植物油菜素文献速览 2022-08-01

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Nat Plants , IF:15.793 , 2022 Jun , V8 (6) : P646-655 doi: 10.1038/s41477-022-01167-1

Deconvoluting signals downstream of growth and immune receptor kinases by phosphocodes of the BSU1 family phosphatases.

Park, Chan Ho and Bi, Yang and Youn, Ji-Hyun and Kim, So-Hee and Kim, Jung-Gun and Xu, Nicole Y and Shrestha, Ruben and Burlingame, Alma L and Xu, Shou-Ling and Mudgett, Mary Beth and Kim, Seong-Ki and Kim, Tae-Wuk and Wang, Zhi-Yong

Department of Plant Biology, Carnegie Institution for Science, Stanford, CA, USA.; Department of Biology, Stanford University, Stanford, CA, USA.; Department of Life Science, College of Natural Sciences, Chung-Ang University, Seoul, Republic of Korea.; Department of Life Science, College of Natural Sciences, Hanyang University, Seoul, Republic of Korea.; Department of Pharmaceutical Chemistry, University of California at San Francisco, San Francisco, CA, USA.; Department of Life Science, College of Natural Sciences, Chung-Ang University, Seoul, Republic of Korea. skkimbio@cau.ac.kr.; Department of Life Science, College of Natural Sciences, Hanyang University, Seoul, Republic of Korea. twgibio@hanyang.ac.kr.; Research Institute for Convergence of Basic Science, Hanyang University, Seoul, South Korea. twgibio@hanyang.ac.kr.; Department of Plant Biology, Carnegie Institution for Science, Stanford, CA, USA. zywang24@stanford.edu.

Hundreds of leucine-rich repeat receptor kinases (LRR-RKs) have evolved to control diverse processes of growth, development and immunity in plants, but the mechanisms that link LRR-RKs to distinct cellular responses are not understood. Here we show that two LRR-RKs, the brassinosteroid hormone receptor BRASSINOSTEROID INSENSITIVE 1 (BRI1) and the flagellin receptor FLAGELLIN SENSING 2 (FLS2), regulate downstream glycogen synthase kinase 3 (GSK3) and mitogen-activated protein (MAP) kinases, respectively, through phosphocoding of the BRI1-SUPPRESSOR1 (BSU1) phosphatase. BSU1 was previously identified as a component that inactivates GSK3s in the BRI1 pathway. We surprisingly found that the loss of the BSU1 family phosphatases activates effector-triggered immunity and impairs flagellin-triggered MAP kinase activation and immunity. The flagellin-activated BOTRYTIS-INDUCED KINASE 1 (BIK1) phosphorylates BSU1 at serine 251. Mutation of serine 251 reduces BSU1's ability to mediate flagellin-induced MAP kinase activation and immunity, but not its abilities to suppress effector-triggered immunity and interact with GSK3, which is enhanced through the phosphorylation of BSU1 at serine 764 upon brassinosteroid signalling. These results demonstrate that BSU1 plays an essential role in immunity and transduces brassinosteroid-BRI1 and flagellin-FLS2 signals using different phosphorylation sites. Our study illustrates that phosphocoding in shared downstream components provides signalling specificities for diverse plant receptor kinases.

PMID: 35697730


Mol Plant , IF:13.164 , 2022 Jun , V15 (6) : P991-1007 doi: 10.1016/j.molp.2022.05.002

Brassinosteroids enhance salicylic acid-mediated immune responses by inhibiting BIN2 phosphorylation of clade I TGA transcription factors in Arabidopsis.

Kim, Yeong-Woo and Youn, Ji-Hyun and Roh, Jeehee and Kim, Jeong-Mok and Kim, Seong-Ki and Kim, Tae-Wuk

Department of Life Science, Hanyang University, Seoul 04763, Republic of Korea.; Department of Life Science, Chung-Ang University, Seoul 06973, Republic of Korea.; Department of Life Science, Chung-Ang University, Seoul 06973, Republic of Korea. Electronic address: skkimbio@cau.ac.kr.; Department of Life Science, Hanyang University, Seoul 04763, Republic of Korea; Research Institute for Convergence of Basic Science, Hanyang University, Seoul 04763, Republic of Korea; Hanyang Institute of Bioscience and Biotechnology, Hanyang University, Seoul 04763, Republic of Korea. Electronic address: twgibio@hanyang.ac.kr.

Salicylic acid (SA) plays an important role in plant immune response, including resistance to pathogens and systemic acquired resistance. Two major components, NONEXPRESSOR OF PATHOGENESIS-RELATED GENES (NPRs) and TGACG motif-binding transcription factors (TGAs), are known to mediate SA signaling, which might also be orchestrated by other hormonal and environmental changes. Nevertheless, the molecular and functional interactions between SA signaling components and other cellular signaling pathways remain poorly understood. Here we showed that the steroid plant hormone brassinosteroid (BR) promotes SA responses by inactivating BR-INSENSITIVE 2 (BIN2), which inhibits the redox-sensitive clade I TGAs in Arabidopsis. We found that both BR and the BIN2 inhibitor bikinin synergistically increase SA-mediated physiological responses, such as resistance to Pst DC3000. Our genetic and biochemical analyses indicated that BIN2 functionally interacts with TGA1 and TGA4, but not with other TGAs. We further demonstrated that BIN2 phosphorylates Ser-202 of TGA4, resulting in the suppression of the redox-dependent interaction between TGA4 and NPR1 as well as destabilization of TGA4. Consistently, transgenic Arabidopsis overexpressing TGA4-YFP with a S202A mutation displayed enhanced SA responses compared to the wild-type TGA4-YFP plants. Taken together, these results suggest a novel crosstalk mechanism by which BR signaling coordinates the SA responses mediated by redox-sensitive clade I TGAs.

PMID: 35524409


Plant Cell , IF:11.277 , 2022 Jul doi: 10.1093/plcell/koac203

Adenosine monophosphate deaminase modulates BIN2 activity through hydrogen peroxide-induced oligomerization.

Lu, Qing and Houbaert, Anaxi and Ma, Qian and Huang, Jingjing and Sterck, Lieven and Zhang, Cheng and Benjamins, Rene and Coppens, Frederik and Van Breusegem, Frank and Russinova, Eugenia

Department of Plant Biotechnology and Bioinformatics, Ghent University, 9052 Ghent, Belgium.; Center for Plant Systems Biology, VIB, 9052 Ghent, Belgium.; Plant Developmental Biology, Wageningen University Research, 6708 PB Wageningen, The Netherlands.

The Arabidopsis thaliana GSK3-like kinase, BRASSINOSTEROID-INSENSITIVE2 (BIN2) is a key negative regulator of brassinosteroid (BR) signaling and a hub for crosstalk with other signaling pathways. However, the mechanisms controlling BIN2 activity are not well understood. Here we performed a forward genetic screen for resistance to the plant-specific GSK3 inhibitor bikinin and discovered that a mutation in the ADENOSINE MONOPHOSPHATE DEAMINASE (AMPD)/EMBRYONIC FACTOR1 (FAC1) gene reduces the sensitivity of Arabidopsis seedlings to both bikinin and BRs. Further analyses showed that AMPD modulates BIN2 activity by regulating its oligomerization in a hydrogen peroxide (H2O2)-dependent manner. Exogenous H2O2 induced the formation of BIN2 oligomers with a decreased kinase activity and an increased sensitivity to bikinin. By contrast, AMPD activity inhibition reduces the cytosolic reactive oxygen species (ROS) levels and the amount of BIN2 oligomers, correlating with the decreased sensitivity of Arabidopsis plants to bikinin and BRs. Furthermore, we showed that BIN2 phosphorylates AMPD to possibly alter its function. Our results reveal the existence of a H2O2 homeostasis-mediated regulation loop between AMPD and BIN2 that fine-tunes the BIN2 kinase activity to control plant growth and development.

PMID: 35876813


Plant Cell , IF:11.277 , 2022 Jul doi: 10.1093/plcell/koac196

Rice DWARF AND LOW-TILLERING and the homeodomain protein OSH15 interact to regulate internode elongation via orchestrating brassinosteroid signaling and metabolism.

Niu, Mei and Wang, Hongru and Yin, Wenchao and Meng, Wenjing and Xiao, Yunhua and Liu, Dapu and Zhang, Xiaoxing and Dong, Nana and Liu, Jihong and Yang, Yanzhao and Zhang, Fan and Chu, Chengcai and Tong, Hongning

National Key Facility for Crop Gene Resources and Genetic Improvement, Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, China.; State Key Laboratory of Plant Genomics and Center for Plant Gene Research (Beijing), Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China.; Southern Regional Collaborative Innovation Center for Grain and Oil Crops in China, Hunan Agricultural University, Changsha 410128, China.; National Nanfan Research Institute, Chinese Academy of Agricultural Sciences, Sanya 572024, China.

Brassinosteroid (BR) phytohormones play crucial roles in regulating internode elongation in rice (Oryza sativa). However, the underlying mechanism remains largely unclear. The dwarf and low-tillering (dlt) mutant is a mild BR-signaling-defective mutant. Here we identify two dlt enhancers that show more severe shortening of the lower internodes compared to the uppermost internode (IN1). Both mutants carry alleles of ORYZA SATIVA HOMEOBOX 15 (OSH15), the founding gene for dwarf6-type mutants, which have shortened lower internodes but not IN1. Consistent with the mutant phenotype, OSH15 expression is much stronger in lower internodes, particularly in IN2, than IN1. The osh15 single mutants have impaired BR sensitivity accompanied by enhanced BR synthesis in seedlings. DLT physically interacts with OSH15 to co-regulate many genes in seedlings and internodes. OSH15 targets and promotes expression of the BR receptor gene BR INSENSITIVE1 (OsBRI1), and DLT facilitates this regulation in a dosage-dependent manner. In osh15, dlt and osh15 dlt, BR levels are higher in seedlings and panicles, but unexpectedly lower in internodes compared with wild type. Taken together, our results suggest that DLT interacts with OSH15, which functions in the lower internodes, to modulate rice internode elongation via orchestrating BR signaling and metabolism.

PMID: 35789396


Curr Biol , IF:10.834 , 2022 Jun , V32 (11) : P2454-2466.e7 doi: 10.1016/j.cub.2022.04.028

The receptor kinase OsWAK11 monitors cell wall pectin changes to fine-tune brassinosteroid signaling and regulate cell elongation in rice.

Yue, Zhi-Liang and Liu, Ning and Deng, Zhi-Ping and Zhang, Yu and Wu, Zhi-Ming and Zhao, Ji-Long and Sun, Ying and Wang, Zhi-Yong and Zhang, Sheng-Wei

Ministry of Education Key Laboratory of Molecular and Cellular Biology, Hebei Collaboration Innovation Center for Cell Signaling and Environmental Adaptation, Hebei Key Laboratory of Molecular and Cellular Biology, College of Life Sciences, Hebei Normal University, 050024 Shijiazhuang, China; Institute of Cash Crops, Hebei Academy of Agriculture & Forestry Sciences, Shijiazhuang 050051, China.; Ministry of Education Key Laboratory of Molecular and Cellular Biology, Hebei Collaboration Innovation Center for Cell Signaling and Environmental Adaptation, Hebei Key Laboratory of Molecular and Cellular Biology, College of Life Sciences, Hebei Normal University, 050024 Shijiazhuang, China.; State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-products, Institute of Virology and Biotechnology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China.; Institute of Cash Crops, Hebei Academy of Agriculture & Forestry Sciences, Shijiazhuang 050051, China.; Department of Plant Biology, Carnegie Institution for Science, Stanford, CA 94305, USA.; Ministry of Education Key Laboratory of Molecular and Cellular Biology, Hebei Collaboration Innovation Center for Cell Signaling and Environmental Adaptation, Hebei Key Laboratory of Molecular and Cellular Biology, College of Life Sciences, Hebei Normal University, 050024 Shijiazhuang, China. Electronic address: swzhang@hebtu.edu.cn.

Rates of plant cell elongation change with day-night alternation, reflecting differences in metabolism related to cell wall remodeling. Information from cell wall surveillance pathways must be integrated with growth regulation pathways to provide feedback regulation of cell wall modification; such feedback regulation is important to ensure sufficient strength and prevent rupture of the cell wall during growth. Several lines of evidence suggest that cell wall perturbations often influence phytohormone signaling, but the identity of the nexus between these two processes remained elusive. Here, we show that wall-associated kinase11 (OsWAK11) acts as a linker connecting cell wall pectin methyl-esterification changes and brassinosteroid (BR) signaling in rice. Our data show that OsWAK11 controls several important agronomical traits by regulating cell elongation in rice. OsWAK11 directly binds and phosphorylates the BR receptor OsBRI1 at residue Thr752, within a motif conserved across most monocot graminaceous crops, thus hindering OsBRI1 interaction with its co-receptor OsSERK1/OsBAK1 and inhibiting BR signaling. The extracellular domain of OsWAK11 shows a much stronger interaction toward methyl-esterified pectin as compared with de-methyl-esterified pectin. OsWAK11 is stabilized in light but is degraded in darkness, in a process triggered by changes in the ratio of methyl-esterified to de-methyl-esterified pectin, creating fluctuations in plant BR signaling in response to day and night alternation. We conclude that OsWAK11 is a cell wall monitor that regulates cell elongation rates to adapt to the environment from the outside in, which complements the well-established inside-out signaling pathway affecting cell elongation in plants.

PMID: 35512695


New Phytol , IF:10.151 , 2022 Jul doi: 10.1111/nph.18404

Integration of multi-omics data reveals interplay between brassinosteroid and TORC signaling in Arabidopsis.

Montes, Christian and Wang, Ping and Liao, Ching-Yi and Nolan, Trevor M and Song, Gaoyuan and Clark, Natalie M and Elmore, J Mitch and Guo, Hongqing and Bassham, Diane C and Yin, Yanhai and Walley, Justin W

Department of Plant Pathology and Microbiology, Iowa State University, Ames, IA, 50011, USA.; Department of Genetics, Development and Cell Biology, Iowa State University, Ames, IA, 50011, USA.; Department of Biology, Duke University, Durham, NC, 27708, USA.; USDA-ARS Cereal Disease Laboratory, University of Minnesota, St. Paul, MN, 55108, USA.; Plant Sciences Institute, Iowa State University, Ames, IA, 50011, USA.

Brassinosteroids (BR) and Target of Rapamycin Complex (TORC) are two major actors coordinating plant growth and stress responses. BRs function through a signaling pathway to extensively regulate gene expression and TORC is known to regulate translation and autophagy. Recent studies have revealed connections between these two pathways, but a system-wide view of their interplay is still missing.We quantified the level of 23,975 transcripts, 11,183 proteins, and 27,887 phosphorylation sites in wild-type Arabidopsis thaliana and in mutants with altered levels of either BRASSINOSTEROID INSENSITIVE 2 (BIN2) or REGULATORY ASSOCIATED PROTEIN OF TOR 1B (RAPTOR1B), two key players in BR and TORC signaling, respectively.We found that perturbation of BIN2 or RAPTOR1B levels affects a common set of gene-products involved in growth and stress responses. Furthermore, we used the multi-omic data to reconstruct an integrated signaling network. We screened 41 candidate genes identified from the reconstructed network and found that loss of function mutants of many of these proteins led to an altered BR response and/or modulated autophagy activity.Altogether, these results establish a predictive network that defines different layers of molecular interactions between BR- or TORC-regulated growth and autophagy.

PMID: 35892179


New Phytol , IF:10.151 , 2022 Aug , V235 (4) : P1455-1469 doi: 10.1111/nph.18228

Pan-brassinosteroid signaling revealed by functional analysis of NILR1 in land plants.

Zheng, Bowen and Bai, Qunwei and Li, Chenxi and Wang, Lihaitian and Wei, Qiang and Ali, Khawar and Li, Wenjuan and Huang, Shengdi and Xu, Hongxing and Li, Guishuang and Ren, Hongyan and Wu, Guang

College of Life Sciences, Shaanxi Normal University, Xi'an, 710119, China.

Brassinosteroid (BR) signaling has been identified from the ligand BRs sensed by the receptor Brassinosteroid Insensitive 1 (BRI1) to the final activation of Brassinozole Resistant 1/bri1 EMS-Suppressor 1 through a series of transduction events. Extensive studies have been conducted to characterize the role of BR signaling in various biological processes. Our previous study has shown that Excess Microsporocytes 1 (EMS1) and BRI1 control different aspects of plant growth and development via conserved intracellular signaling. Here, we reveal that another receptor, NILR1, can complement the bri1 mutant in the absence of BRs, indicating a pathway that resembles BR signaling activated by NILR1. Genetic analysis confirms the intracellular domains of NILR1, BRI1 and EMS1 have a common signal output. Furthermore, we demonstrate that NILR1 and BRI1 share the coreceptor BRI1 Associated Kinase 1 and substrate BSKs. Notably, the NILR1-mediated downstream pathway is conserved across land plants. In summary, we provide evidence for the signaling cascade of NILR1, suggesting pan-brassinosteroid signaling initiated by a group of distant receptor-ligand pairs in land plants.

PMID: 35570834


Plant Physiol , IF:8.34 , 2022 Jul doi: 10.1093/plphys/kiac354

DIACYLGLYCEROL KINASE 5 Participates in Flagellin-Induced Signaling in Arabidopsis.

Kalachova, Tetiana and Skrabalkova, Eliska and Pateyron, Stephanie and Soubigou-Taconnat, Ludivine and Djafi, Nabila and Collin, Sylvie and Sekeres, Juraj and Burketova, Lenka and Potocky, Martin and Pejchar, Premysl and Ruelland, Eric

Institute of Experimental Botany of the Czech Academy of Sciences, Rozvojova 263, 16502 Prague, Czech Republic.; Department of Experimental Plant Biology, Charles University, Vinicna 5, Prague 12844, Czech Republic.; Institute of Plant Sciences Paris Saclay IPS2, CNRS, INRA, Universite Paris-Sud, Universite Evry, Universite Paris-Saclay, Batiment 630, 91405 Orsay, France.; Institute of Plant Sciences Paris-Saclay IPS2, Paris Diderot, Sorbonne Paris-Cite, Batiment 630, 91405, Orsay, France.; Sorbonne Universite, Physiologie Cellulaire et Moleculaire des Plantes, F-75005 Paris, France.; Universite de Technologie de Compiegne, CNRS Enzyme and Cell Engineering Laboratory, Rue du Docteur Schweitzer, 60203 Compiegne, France.

Flagellin perception is a keystone of pattern-triggered immunity in plants. The recognition of this protein by a plasma membrane receptor complex is the beginning of a signaling cascade that includes protein phosphorylation and the production of reactive oxygen species (ROS). In both Arabidopsis (Arabidopsis thaliana) seedlings and suspension cells, we found that treatment with flg22, a peptide corresponding to the most conserved domain of bacterial flagellin, caused a rapid and transient decrease in the level of phosphatidylinositol 4,5-bisphosphate along with a parallel increase in phosphatidic acid (PA). In suspension cells, inhibitors of either phosphoinositide-dependent phospholipases C (PLC) or diacylglycerol kinases (DGKs) inhibited flg22-triggered PA production and the oxidative burst. In response to flg22, receptor-like kinase-deficient fls2, bak1 and bik1 mutants (FLAGELLIN SENSITIVE 2, BRASSINOSTEROID INSENSITIVE 1-associated kinase 1 and BOTRYTIS-INDUCED KINASE 1, respectively) produced less PA than wild-type (WT) plants, whereas this response did not differ in NADPH oxidase-deficient rbohD (RESPIRATORY BURST OXIDASE HOMOLOG D) plants. Among the DGK-deficient lines tested, the dgk5.1 mutant produced less PA and less ROS after flg22 treatment compared to WT seedlings. In response to flg22, dgk5.1 plants showed lower callose accumulation and impaired resistance to Pseudomonas syringae pv. tomato DC3000 hrcC-. Transcriptomics revealed that the basal expression of defense-related genes was altered in dgk5.1 seedlings compared to the WT. A GFP-DGK5 fusion protein localized to the plasma membrane, where RBOHD and PLC2 (proteins involved in plant immunity) are also located. The role of DGK5 and its enzymatic activity in flagellin signaling and fine-tuning of early immune responses in plant-microbe interactions is discussed.

PMID: 35900211


Plant Physiol , IF:8.34 , 2022 Jul doi: 10.1093/plphys/kiac332

Class I TCP transcription factor AtTCP8 modulates key brassinosteroid-responsive genes.

Spears, Benjamin J and McInturf, Samuel A and Collins, Carina and Chlebowski, Meghann and Cseke, Leland J and Su, Jianbin and Mendoza-Cozatl, David G and Gassmann, Walter

Department of Biological Sciences, Butler University, Indianapolis, Indiana, United States of America.; Division of Plant Sciences, University of Missouri, Columbia, Missouri, United States of America.; Christopher S. Bond Life Sciences Center and Interdisciplinary Plant Group, University of Missouri, Columbia, Missouri, United States of America.; Department of Biology, Marian University, Indianapolis, Indiana, United States of America.

The plant-specific TEOSINTE BRANCHED1/CYCLOIDEA/PROLIFERATING CELL FACTOR (TCP) transcription factor family is most closely associated with regulating plant developmental programs. Recently, TCPs were also shown to mediate host immune signaling, both as targets of pathogen virulence factors and as regulators of plant defense genes. However, comprehensive characterization of TCP gene targets is still lacking. Loss of function of the class I TCP gene AtTCP8 attenuates early immune signaling and, when combined with mutations in AtTCP14 and AtTCP15, additional layers of defense signaling in Arabidopsis (Arabidopsis thaliana). Here, we focus on TCP8, the most poorly characterized of the three to date. We used chromatin immunoprecipitation and RNA-sequencing to identify TCP8-bound gene promoters and differentially regulated genes in the tcp8 mutant; these data sets were heavily enriched in signaling components for multiple phytohormone pathways, including brassinosteroids (BRs), auxin, and jasmonic acid. Using BR signaling as a representative example, we showed that TCP8 directly binds and activates the promoters of the key BR transcriptional regulatory genes BRASSINAZOLE-RESISTANT1 (BZR1) and BRASSINAZOLE-RESISTANT2 (BZR2/BES1). Furthermore, tcp8 mutant seedlings exhibited altered BR-responsive growth patterns and complementary reductions in BZR2 transcript levels, while TCP8 protein demonstrated BR-responsive changes in subnuclear localization and transcriptional activity. We conclude that one explanation for the substantial targeting of TCP8 alongside other TCP family members by pathogen effectors may lie in its role as a modulator of BR and other plant hormone signaling pathways.

PMID: 35866682


Plant Physiol , IF:8.34 , 2022 Jul doi: 10.1093/plphys/kiac327

Brassinosteroid signaling restricts root lignification by antagonizing SHORT-ROOT function in Arabidopsis.

Li, Meng and Li, Pengxue and Wang, Chunhua and Xu, Huimin and Wang, Mengxue and Wang, Yanli and Niu, Xufang and Xu, Mengyuan and Wang, Hong and Qin, Yaxin and Tang, Wenqiang and Bai, Mingyi and Wang, Wenfei and Wu, Shuang

College of Life Science&College of Horticulture, Fujian Agriculture and Forestry University, Fuzhou 350002, China.; Ministry of Education Key Laboratory of Molecular and Cellular Biology, Hebei Collaboration Innovation Center for Cell Signaling, Hebei Key Laboratory of Molecular and Cellular Biology, College of Life Sciences, Hebei Normal University, Shijiazhuang, China.; Key Laboratory of Plant Development and Environment Adaptation Biology, Ministry of Education, School of Life Sciences, Shandong University, Jinan, China.

Cell wall lignification is a key step in forming functional endodermis and protoxylem in plant roots. Lignified casparian strips (CS) in endodermis and tracheary elements of protoxylem are essential for selective absorption and transport of water and nutrients. Although multiple key regulators of CS and protoxylem have been identified, the spatial information that drives the developmental shift to root lignification remains unknown. Here, we found that brassinosteroid signaling plays a key role in inhibiting root lignification in the root elongation zone. The inhibitory activity of brassinosteroid signaling occurs partially through the direct binding of BRASSINAZOLE-RESISTANT 1 (BZR1) to SHORT-ROOT (SHR), repressing the SHR-mediated activation of downstream genes that are involved in root lignification. Upon entering the mature root zone, brassinosteroid signaling declines rapidly, which releases SHR activity and initiates root lignification. Our results provide a mechanistic view of the developmental transition to cell wall lignification in Arabidopsis thaliana roots.

PMID: 35809074


Plant Physiol , IF:8.34 , 2022 Jun , V189 (3) : P1177-1179 doi: 10.1093/plphys/kiac185

Transcription factor OsNAC016: a convergent point of brassinosteroid and abscisic acid signaling in rice.

Ahmed, Sulaiman and Chen, Jian

International Genome Center, Jiangsu University, Zhenjiang 212013, China.

PMID: 35460247


Plant Physiol , IF:8.34 , 2022 Jun , V189 (3) : P1757-1773 doi: 10.1093/plphys/kiac157

The interplay of auxin and brassinosteroid signaling tunes root growth under low and different nitrogen forms.

Devi, Loitongbam Lorinda and Pandey, Anshika and Gupta, Shreya and Singh, Amar Pal

National Institute of Plant Genome Research, New Delhi, 110067, India.

The coordinated signaling activity of auxin and brassinosteroids (BRs) is critical for optimal plant growth and development. Nutrient-derived signals regulate root growth by modulating the levels and spatial distribution of growth hormones to optimize nutrient uptake and assimilation. However, the effect of the interaction of these two hormones and their signaling on root plasticity during low and differential availability of nitrogen (N) forms (NH4+/NO3-) remains elusive. We demonstrate that root elongation under low N (LN) is an outcome of the interdependent activity of auxin and BR signaling pathways in Arabidopsis (Arabidopsis thaliana). LN promotes root elongation by increasing BR-induced auxin transport activity in the roots. Increased nuclear auxin signaling and its transport efficiency have a distinct impact on root elongation under LN conditions. High auxin levels reversibly inhibit BR signaling via BRI1 KINASE INHIBITOR1. Using the tissue-specific approach, we show that BR signaling from root vasculature (stele) tissues is sufficient to promote cell elongation and, hence, root growth under LN condition. Further, we show that N form-defined root growth attenuation or enhancement depends on the fine balance of BR and auxin signaling activity. NH4+ as a sole N source represses BR signaling and response, which in turn inhibits auxin response and transport, whereas NO3- promotes root elongation in a BR signaling-dependent manner. In this study, we demonstrate the interplay of auxin and BR-derived signals, which are critical for root growth in a heterogeneous N environment and appear essential for root N foraging response and adaptation.

PMID: 35377445


Plant Physiol , IF:8.34 , 2022 Jun , V189 (3) : P1296-1313 doi: 10.1093/plphys/kiac146

OsNAC016 regulates plant architecture and drought tolerance by interacting with the kinases GSK2 and SAPK8.

Wu, Qi and Liu, Yingfan and Xie, Zizhao and Yu, Bo and Sun, Ying and Huang, Junli

Key Laboratory of Biorheological Science and Technology, Ministry of Education, Bioengineering College, Chongqing University, Chongqing 400044, China.

Ideal plant architecture and drought tolerance are important determinants of yield potential in rice (Oryza sativa). Here, we found that OsNAC016, a rice NAC (NAM, ATAF, and CUC) transcription factor, functions as a regulator in the crosslink between brassinosteroid (BR)-mediated plant architecture and abscisic acid (ABA)-regulated drought responses. The loss-of-function mutant osnac016 exhibited erect leaves and shortened internodes, but OsNAC016-overexpressing plants had opposite phenotypes. Further investigation revealed that OsNAC016 regulated the expression of the BR biosynthesis gene D2 by binding to its promoter. Moreover, OsNAC016 interacted with and was phosphorylated by GSK3/SHAGGY-LIKE KINASE2 (GSK2), a negative regulator in the BR pathway. Meanwhile, the mutant osnac016 had improved drought stress tolerance, supported by a decreased water loss rate and enhanced stomatal closure in response to exogenous ABA, but OsNAC016-overexpressing plants showed attenuated drought tolerance and reduced ABA sensitivity. Further, OSMOTIC STRESS/ABA-ACTIVATED PROTEIN KINASE8 (SAPK8) phosphorylated OsNAC016 and reduced its stability. The ubiquitin/26S proteasome system is an important degradation pathway of OsNAC016 via the interaction with PLANT U-BOX PROTEIN43 (OsPUB43) that mediates the ubiquitination of OsNAC016. Notably, RNA-sequencing analysis revealed global roles of OsNAC016 in promoting BR-mediated gene expression and repressing ABA-dependent drought-responsive gene expression, which was confirmed by chromatin immunoprecipitation quantitative PCR analysis. Our findings establish that OsNAC016 is positively involved in BR-regulated rice architecture, negatively modulates ABA-mediated drought tolerance, and is regulated by GSK2, SAPK8, and OsPUB43 through posttranslational modification. Our data provide insights into how plants balance growth and survival by coordinately regulating the growth-promoting signaling pathway and response under abiotic stresses.

PMID: 35333328


Sci Total Environ , IF:7.963 , 2022 Sep , V838 (Pt 4) : P156503 doi: 10.1016/j.scitotenv.2022.156503

OsBR6ox, a member in the brassinosteroid synthetic pathway facilitates degradation of pesticides in rice through a specific DNA demethylation mechanism.

Qiao, Yuxin and Ma, Li Ya and Chen, Zhao Jie and Wang, Yujue and Gu, Yucheng and Yang, Hong

Jiangsu Key Laboratory of Pesticide Science, College of Sciences, Nanjing Agricultural University, Nanjing 210095, China.; Jiangsu Key Laboratory of Pesticide Science, College of Sciences, Nanjing Agricultural University, Nanjing 210095, China; Institute of Agricultural Resources and Environment, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China.; Syngenta Crop Protection AG, Rosentalstrasse 67, CH-4002 Basel, Switzerland.; Syngenta Ltd, Jealott's Hill International Research Centre, Bracknell, Berkshire RG42 6EY, UK.; Jiangsu Key Laboratory of Pesticide Science, College of Sciences, Nanjing Agricultural University, Nanjing 210095, China. Electronic address: hongyang@njau.edu.cn.

This manuscript described a comprehensive study on a pesticide degradation factor OsBR6ox that promoted the degradation of pesticides atrazine (ATZ) and acetochlor (ACT) in rice tissues and grains through an epigenetic mechanism. OsBR6ox was transcriptionally induced under ATZ and ACT stress. Genetic disruption of OsBR6ox increased rice sensitivity and led to more accumulation of ATZ and ACT, whereas transgenic rice overexpressing OsBR6ox lines (OEs) showed opposite effects with improved growth and lower ATZ and ACT accumulation in various tissues, including grains. OsBR6ox-mediated detoxification of ATZ and ACT was associated with the increased abundance of brassinolide (one of the brassinosteroids, BRs), a plant growth regulator for stress responses. Some Phase I-II reaction protein genes for pesticide detoxification such as genes encoding laccase, O-methyltransferase and glycosyltransferases were transcriptionally upregulated in OE lines under ATZ and ACT stress. HPLC-Q-TOF-MS/MS analysis revealed an enhanced ATZ/ATC metabolism in OE plants, which removed 1.21-1.49 fold ATZ and 1.31-1.44 fold ACT from the growth medium but accumulated only 83.1-87.1 % (shoot) and 71.7-84.1 % (root) of ATZ and 69.4-83.4 % of ACT of the wild-type. Importantly, an ATZ-responsive demethylated region in the upstream of OsBR6ox was detected. Such an epigenetic modification marker was responsible for the increased OsBR6ox expression and consequent detoxification of ATZ/ACT in rice and environment. Overall, this work uncovered a new model showing that plants utilize two mechanisms to co-regulate the detoxification and metabolism of pesticides in rice and provided a new approach for building up cleaner crops and eliminating residual pesticides in environments.

PMID: 35688248


J Integr Plant Biol , IF:7.061 , 2022 Jul doi: 10.1111/jipb.13327

The RECEPTOR-LIKE PROTEIN53 immune complex associates with LLG1 to positively regulate plant immunity.

Chen, Renjie and Sun, Pengwei and Zhong, Guitao and Wang, Wei and Tang, Dingzhong

State Key Laboratory of Ecological Control of Fujian-Taiwan Crop Pests, Key Laboratory of Ministry of Education for Genetics, Breeding and Multiple Utilization of Crops, Plant Immunity Center, Fujian Agriculture and Forestry University, Fuzhou, 350002, China.; Institute of Genetics and Developmental Biology, the Chinese Academy of Sciences, Beijing, 100101, China.; College of Life Sciences, University of Chinese Academy of Sciences, Beijing, 100049, China.; Ministerial and Provincial Joint Innovation Centre for Safety Production of Cross-Strait Crops, Fujian Agriculture and Forestry University, Fuzhou, 350002, China.

Pattern recognition receptors (PRRs) sense ligands in pattern-triggered immunity (PTI). Plant PRRs include numerous receptor-like proteins (RLPs), but many RLPs remain functionally uncharacterized. Here, we examine an Arabidopsis thaliana RLP, RLP53, which positively regulates immune signaling. Our forward genetic screen for suppressors of enhanced disease resistance1 (edr1) identified a point mutation in RLP53 that fully suppresses disease resistance and mildew-induced cell death in edr1 mutants. The rlp53 mutants showed enhanced susceptibility to virulent pathogens, including fungi, oomycetes, and bacteria, indicating that RLP53 is important for plant immunity. The ectodomain of RLP53 contains leucine-rich repeat (LRR) motifs. RLP53 constitutively associates with the LRR receptor-like kinase SUPPRESSOR OF BRASSINOSTEROID INSENSITIVE1-ASSOCIATED KINASE (BAK1)-INTERACTING RECEPTOR KINASE1 (SOBIR1) and interacts with the co-receptor BAK1 in a pathogen-induced manner. The double mutation sobir1-12 bak1-5 suppresses edr1-mediated disease resistance, suggesting that EDR1 negatively regulates PTI modulated by the RLP53-SOBIR1-BAK1 complex. Moreover, the glycosylphosphatidylinositol (GPI)-anchored protein LORELEI-LIKE GPI-ANCHORED PROTEIN1 (LLG1) interacts with RLP53 and mediates RLP53 accumulation in the plasma membrane. We thus uncovered the role of a novel RLP and its associated immune complex in plant defense responses and revealed a potential new mechanism underlying regulation of RLP immune function by a GPI-anchored protein. This article is protected by copyright. All rights reserved.

PMID: 35796320


J Integr Plant Biol , IF:7.061 , 2022 Jun doi: 10.1111/jipb.13322

The divergence of brassinosteroid sensitivity between rice subspecies involves natural variation conferring altered internal auto-binding of OsBSK2.

Yin, Wenchao and Li, Lulu and Yu, Zhikun and Zhang, Fan and Liu, Dapu and Wu, Hongkai and Niu, Mei and Meng, Wenjing and Zhang, Xiaoxing and Dong, Nana and Yang, Yanzhao and Liu, Jihong and Liu, Yongqiang and Zhang, Guoxia and Xu, Jianlong and Wang, Shimei and Chu, Chengcai and Qian, Qian and Tong, Hongning

National Key Facility for Crop Gene Resources and Genetic Improvement, Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing, 100081, China.; Rice Research Institute of Anhui Academy of Agricultural Sciences, Hefei, 230001, China.; State Key Laboratory of Plant Genomics, Institute of Genetics and Developmental Biology, the Innovative Academy for Seed Design, Chinese Academy of Sciences, Beijing, 100101, China.

Japonica/geng and indica/xian are two major rice (Oryza sativa) subspecies with multiple divergent traits, but how these traits are related and interact within each subspecies remains elusive. Brassinosteroids (BRs) are a class of steroid phytohormones that modulate many important agronomic traits in rice. Here, using different physiological assays, we revealed that japonica rice exhibits an overall lower BR sensitivity than indica. Extensive screening of BR signaling genes led to the identification of a set of genes distributed throughout the primary BR signaling pathway with divergent polymorphisms. Among these, we demonstrate that the C38/T variant in BR Signaling Kinase2 (OsBSK2), causing the amino acid change P13L, plays a central role in mediating differential BR signaling in japonica and indica rice. OsBSK2(L13) in indica plays a greater role in BR signaling than OsBSK2(P13) in japonica by affecting the auto-binding and protein accumulation of OsBSK2. Finally, we determined that OsBSK2 is involved in a number of divergent traits in japonica relative to indica rice, including grain shape, tiller number, cold adaptation, and nitrogen-use efficiency. Our study suggests that the natural variation in OsBSK2 plays a key role in the divergence of BR signaling, which underlies multiple divergent traits between japonica and indica.

PMID: 35766344


J Integr Plant Biol , IF:7.061 , 2022 Jun doi: 10.1111/jipb.13311

OsCPL3 is involved in brassinosteroid signaling by regulating OsGSK2 stability.

Gong, Luping and Liao, Shenghao and Duan, Wen and Liu, Yongqiang and Zhu, Dongmei and Zhou, Xiaosheng and Xue, Baoping and Chu, Chengcai and Liang, Yun-Kuan

State Key Laboratory of Hybrid Rice, Department of Plant Sciences, College of Life Sciences, Wuhan University, Wuhan, 430072, China.; State Key Laboratory of Plant Genomics, Institute of Genetics and Developmental Biology, the Innovative Academy for Seed Design, Chinese Academy of Sciences, Beijing, 100101, China.; Hubei Hongshan Laboratory, Wuhan, 430070, China.

Glycogen synthase kinase 3 (GSK3) proteins play key roles in brassinosteroid (BR) signaling during plant growth and development by phosphorylating various substrates. However, how GSK3 protein stability and activity are themselves modulated is not well understood. Here, we demonstrate in vitro and in vivo that C-TERMINAL DOMAIN PHOSPHATASE-LIKE 3 (OsCPL3), a member of the RNA Pol II CTD phosphatase-like family, physically interacts with OsGSK2 in rice (Oryza sativa). OsCPL3 expression was widely detected in various tissues and organs including roots, leaves and lamina joints, and was induced by exogenous BR treatment. OsCPL3 localized to the nucleus, where it dephosphorylated OsGSK2 at the Ser-222 and Thr-284 residues to modulate its protein turnover and kinase activity, in turn affecting the degradation of BRASSINAZOLE-RESISTANT 1 (BZR1) and BR signaling. Loss of OsCPL3 function resulted in higher OsGSK2 abundance and lower OsBZR1 levels, leading to decreased BR responsiveness and alterations in plant morphology including semi-dwarfism, leaf erectness and grain size, which are of fundamental importance to crop productivity. These results reveal a previously unrecognized role for OsCPL3 and add another layer of complexity to the tightly controlled BR signaling pathway in plants.

PMID: 35665602


Food Res Int , IF:6.475 , 2022 Jul , V157 : P111296 doi: 10.1016/j.foodres.2022.111296

Comparative transcriptomic analysis reveals the potential mechanism of hot water treatment alleviated-chilling injury in banana fruit.

Si, Jia and Fan, Yin-Yin and Liu, Zong-Li and Wei, Wei and Xiao, Xiao-Mei and Yang, Ying-Ying and Shan, Wei and Kuang, Jian-Fei and Lu, Wang-Jin and Fan, Zhong-Qi and Li, Lu-Lu and Chen, Jian-Ye

State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources/Guangdong Provincial Key Laboratory of Postharvest Science of Fruits and Vegetables/Engineering Research Center of Southern Horticultural Products Preservation, Ministry of Education, College of Horticulture, South China Agricultural University, Guangzhou 510642, China.; Key Laboratory of Postharvest Biology of Subtropical Special Agricultural Products/Institute of Postharvest Technology of Agricultural Products, College of Food Science, Fujian Agriculture and Forestry University, Fuzhou, Fujian 350002, China. Electronic address: fanzqi@fafu.edu.cn.; Tea Research Institute, Guizhou Academy of Agricultural Sciences, Guiyang, Guizhou 550006, China.; State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources/Guangdong Provincial Key Laboratory of Postharvest Science of Fruits and Vegetables/Engineering Research Center of Southern Horticultural Products Preservation, Ministry of Education, College of Horticulture, South China Agricultural University, Guangzhou 510642, China. Electronic address: chenjianye@scau.edu.cn.

Banana fruit is prone to chilling injury (CI) during cold storage, resulting in quality deterioration and commodity reduction. The hot water treatment (HWT), dipping banana fruit in hot water (52 degrees C) for 3 min, reduced CI symptom at 7 degrees C storage. The purpose of this study was to investigate the potential molecular mechanism of HWT on the alleviation of CI of postharvest banana fruit. It was found that HWT treatment obviously inhibited the increases in CI index, relative electrolytic leakage, and the contents of malonaldehyde (MDA) and O2(*-), while enhanced proline accumulation. Further transcriptome analysis in the pericarp of banana fruit was evaluated during storage. The results showed that differentially expressed genes (DEGs) in the comparison between control and HWT group were mainly enriched in photosynthesis, chlorophyll metabolism, lipid metabolism, glutathione metabolism, and brassinosteroid and carotenoid biosynthesis. Moreover, transcriptome expression profiles and RT-qPCR analyses exhibited that the corresponding genes involved in these metabolism pathways and heat shock proteins (HSPs) were upregulated by HWT during cold storage. In general, our findings clearly reveal the potential pathways by which HWT alleviates CI in banana fruit, enriching the theoretical basis for the application of hot water to reduce CI in fruits.

PMID: 35761601


Plant J , IF:6.417 , 2022 Aug , V111 (3) : P785-799 doi: 10.1111/tpj.15852

GhBZR3 suppresses cotton fiber elongation by inhibiting very-long-chain fatty acid biosynthesis.

Shi, Zemin and Chen, Xia and Xue, Huidan and Jia, Tingting and Meng, Funing and Liu, Yunfei and Luo, Xiaomin and Xiao, Guanghui and Zhu, Shengwei

Key Laboratory of Plant Molecular Physiology, Institute of Botany, Chinese Academy of Sciences, Beijing, 100093, China.; College of Life Science, University of Chinese Academy of Sciences, Beijing, 100049, China.; School of Food and Biological Engineering, Shaanxi University of Science and Technology, Xi'an, 710021, China.; School of Ecology and Environment, Northwestern Polytechnical University, Xi'an, 710012, China.; College of Life Sciences, Shaanxi Normal University, Xi'an, 710062, China.

The BRASSINAZOLE-RESISTANT (BZR) transcription factor is a core component of brassinosteroid (BR) signaling and is involved in the development of many plant species. BR is essential for the initiation and elongation of cotton fibers. However, the mechanism of BR-regulating fiber development and the function of BZR is poorly understood in Gossypium hirsutum L. (cotton). Here, we identified a BZR family transcription factor protein referred to as GhBZR3 in cotton. Overexpression of GhBZR3 in Arabidopsis caused shorter root hair length, hypocotyl length, and hypocotyl cell length, indicating that GhBZR3 negatively regulates cell elongation. Pathway enrichment analysis from VIGS-GhBZR3 cotton plants found that fatty acid metabolism and degradation might be the regulatory pathway that is primarily controlled by GhBZR3. Silencing GhBZR3 expression in cotton resulted in taller plant height as well as longer fibers. The very-long-chain fatty acid (VLCFA) content was also significantly increased in silenced GhBZR3 plants compared with the wild type. The GhKCS13 promoter, a key gene for VLCFA biosynthesis, contains two GhBZR3 binding sites. The results of yeast one-hybrid, electrophoretic mobility shift, and luciferase assays revealed that GhBZR3 directly interacted with the GhKCS13 promoter to suppress gene expression. Taken together, these results indicate that GhBZR3 negatively regulates cotton fiber development by reducing VLCFA biosynthesis. This study not only deepens our understanding of GhBZR3 function in cotton fiber development, but also highlights the potential of improving cotton fiber length and plant growth using GhBZR3 and its related genes in future cotton breeding programs.

PMID: 35653239


Plant J , IF:6.417 , 2022 Jul , V111 (1) : P183-204 doi: 10.1111/tpj.15786

A lineage-specific arginine in POS1 is required for fruit size control in Physaleae (Solanaceae) via gene co-option.

Wang, Li and Liu, Xueyang and Li, Qiaoru and Xu, Nan and He, Chaoying

State Key Laboratory of Systematic and Evolutionary Botany, Institute of Botany, Chinese Academy of Sciences, Nanxincun 20, Xiangshan, 100093, Beijing, China.; University of Chinese Academy of Sciences, Yuquan Road 19, 100049, Beijing, China.; The Innovative Academy of Seed Design, Chinese Academy of Sciences, Beijing, China.

Solanaceae have important economic value mainly due to their edible fruits. Physalis organ size 1/cytokinin response factor 3 (POS1/CRF3), a unique gene in Solanaceae, is involved in fruit size variation in Physalis but not in Solanum. However, the underlying mechanisms remain elusive. Here, we found that POS1/CRF3 was likely created via the fusion of CRF7 and CRF8 duplicates. Multiple genetic manipulations revealed that only POS1 and Capsicum POS1 (CaPOS1) functioned in fruit size control via the positive regulation of cell expansion. Comparative studies in a phylogenetic framework showed the directional enhancement of POS1-like expression in the flowers and fruits of Physaleae and the specific gain of certain interacting proteins associated with cell expansion by POS1 and CaPOS1. A lineage-specific single nucleotide polymorphism (SNP) caused the 68th amino acid histidine in the POS1 orthologs of non-Physaleae (Nicotiana and Solanum) to change to arginine in Physaleae (Physalis and Capsicum). Substituting the arginine in Physaleae POS1-like by histidine completely abolished their function in the fruits and the protein-protein interaction (PPI) with calreticulin-3. Transcriptomic comparison revealed the potential downstream pathways of POS1, including the brassinosteroid biosynthesis pathway. However, POS1-like may have functioned ancestrally in abiotic stress within Solanaceae. Our work demonstrated that heterometric expression and a SNP caused a single amino acid change to establish new PPIs, which contributed to the co-option of POS1 in multiple regulatory pathways to regulate cell expansion and thus fruit size in Physaleae. These results provide new insights into fruit morphological evolution and fruit yield control.

PMID: 35481627


Int J Mol Sci , IF:5.923 , 2022 Jul , V23 (14) doi: 10.3390/ijms23147915

Impact of Exogenous Application of Potato Virus Y-Specific dsRNA on RNA Interference, Pattern-Triggered Immunity and Poly(ADP-ribose) Metabolism.

Samarskaya, Viktoriya O and Spechenkova, Nadezhda and Markin, Nikolay and Suprunova, Tatyana P and Zavriev, Sergey K and Love, Andrew J and Kalinina, Natalia O and Taliansky, Michael

Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences, 117997 Moscow, Russia.; Doka-Gene Technologies Ltd., 141880 Rogachevo, Russia.; The James Hutton Institute, Invergowrie, Dundee DD2 5DA, UK.; Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, 119991 Moscow, Russia.

In this work we developed and exploited a spray-induced gene silencing (SIGS)-based approach to deliver double-stranded RNA (dsRNA), which was found to protect potato against potato virus Y (PVY) infection. Given that dsRNA can act as a defence-inducing signal that can trigger sequence-specific RNA interference (RNAi) and non-specific pattern-triggered immunity (PTI), we suspected that these two pathways may be invoked via exogeneous application of dsRNA, which may account for the alterations in PVY susceptibility in dsRNA-treated potato plants. Therefore, we tested the impact of exogenously applied PVY-derived dsRNA on both these layers of defence (RNAi and PTI) and explored its effect on accumulation of a homologous virus (PVY) and an unrelated virus (potato virus X, PVX). Here, we show that application of PVY dsRNA in potato plants induced accumulation of both small interfering RNAs (siRNAs), a hallmark of RNAi, and some PTI-related gene transcripts such as WRKY29 (WRKY transcription factor 29; molecular marker of PTI), RbohD (respiratory burst oxidase homolog D), EDS5 (enhanced disease susceptibility 5), SERK3 (somatic embryogenesis receptor kinase 3) encoding brassinosteroid-insensitive 1-associated receptor kinase 1 (BAK1), and PR-1b (pathogenesis-related gene 1b). With respect to virus infections, PVY dsRNA suppressed only PVY replication but did not exhibit any effect on PVX infection in spite of the induction of PTI-like effects in the presence of PVX. Given that RNAi-mediated antiviral immunity acts as the major virus resistance mechanism in plants, it can be suggested that dsRNA-based PTI alone may not be strong enough to suppress virus infection. In addition to RNAi- and PTI-inducing activities, we also showed that PVY-specific dsRNA is able to upregulate production of a key enzyme involved in poly(ADP-ribose) metabolism, namely poly(ADP-ribose) glycohydrolase (PARG), which is regarded as a positive regulator of biotic stress responses. These findings offer insights for future development of innovative approaches which could integrate dsRNA-induced RNAi, PTI and modulation of poly(ADP-ribose) metabolism in a co-ordinated manner, to ensure a high level of crop protection.

PMID: 35887257


Int J Mol Sci , IF:5.923 , 2022 Jun , V23 (12) doi: 10.3390/ijms23126795

Evolutionary Analysis and Functional Identification of Ancient Brassinosteroid Receptors in Ceratopteris richardii.

Zheng, Bowen and Xing, Kaixin and Zhang, Jiaojiao and Liu, Hui and Ali, Khawar and Li, Wenjuan and Bai, Qunwei and Ren, Hongyan

College of Life Sciences, Shaanxi Normal University, Xi'an 710119, China.

Phytohormones play an important role in the adaptive evolution of terrestrial plants. Brassinosteroids (BRs) are essential hormones that regulate multiple aspects of plant growth and development in angiosperms, but the presence of BR signaling in non-seed plants such as ferns remains unknown. Here, we found that BR promotes the growth of Ceratopteris richardii, while the synthetic inhibitor PCZ inhibits the growth. Using full-length transcriptome sequencing, we identified four BRI1-like receptors. By constructing chimeric receptors, we found that the kinase domains of these four receptors could trigger BR downstream signaling. Further, the extracellular domains of two receptors were functionally interchangeable with that of BRI1. In addition, we identified a co-receptor, CtSERK1, that could phosphorylate with CtBRL2s in vitro. Together, these proved the presence of a receptor complex in Ceratopteris richardii that might perceive BR and activate downstream hormone signaling. Our results shed light on the biological and molecular mechanisms of BR signaling in ferns and the role of BR hormone signaling in the adaptive evolution of terrestrial plants.

PMID: 35743240


Front Plant Sci , IF:5.753 , 2022 , V13 : P952246 doi: 10.3389/fpls.2022.952246

A Predominant Role of AtEDEM1 in Catalyzing a Rate-Limiting Demannosylation Step of an Arabidopsis Endoplasmic Reticulum-Associated Degradation Process.

Zhang, Jianjun and Xia, Yang and Wang, Dinghe and Du, Yamin and Chen, Yongwu and Zhang, Congcong and Mao, Juan and Wang, Muyang and She, Yi-Min and Peng, Xinxiang and Liu, Li and Voglmeir, Josef and He, Zuhua and Liu, Linchuan and Li, Jianming

State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, South China Agricultural University, Guangzhou, China.; Guangdong Key Laboratory for Innovative Development and Utilization of Forest Plant Germplasm, College of Forestry and Landscape Architecture, South China Agricultural University, Guangzhou, China.; Department of Molecular, Cellular, and Developmental Biology, University of Michigan, Ann Arbor, MI, United States.; University of Chinese Academy of Sciences, Beijing, China.; The Center of Excellence for Molecular Plant Sciences, Chinese Academy of Sciences, Shanghai, China.; Glycomics and Glycan Bioengineering Research Center, College of Food Science and Technology, Nanjing Agricultural University, Nanjing, China.

Endoplasmic reticulum-associated degradation (ERAD) is a key cellular process for degrading misfolded proteins. It was well known that an asparagine (N)-linked glycan containing a free alpha1,6-mannose residue is a critical ERAD signal created by Homologous to alpha-mannosidase 1 (Htm1) in yeast and ER-Degradation Enhancing alpha-Mannosidase-like proteins (EDEMs) in mammals. An earlier study suggested that two Arabidopsis homologs of Htm1/EDEMs function redundantly in generating such a conserved N-glycan signal. Here we report that the Arabidopsis irb1 (reversal of bri1) mutants accumulate brassinosteroid-insensitive 1-5 (bri1-5), an ER-retained mutant variant of the brassinosteroid receptor BRI1 and are defective in one of the Arabidopsis Htm1/EDEM homologs, AtEDEM1. We show that the wild-type AtEDEM1, but not its catalytically inactive mutant, rescues irb1-1. Importantly, an insertional mutation of the Arabidopsis Asparagine-Linked Glycosylation 3 (ALG3), which causes N-linked glycosylation with truncated glycans carrying a different free alpha1,6-mannose residue, completely nullifies the inhibitory effect of irb1-1 on bri1-5 ERAD. Interestingly, an insertional mutation in AtEDEM2, the other Htm1/EDEM homolog, has no detectable effect on bri1-5 ERAD; however, it enhances the inhibitory effect of irb1-1 on bri1-5 degradation. Moreover, AtEDEM2 transgenes rescued the irb1-1 mutation with lower efficacy than AtEDEM1. Simultaneous elimination of AtEDEM1 and AtEDEM2 completely blocks generation of alpha1,6-mannose-exposed N-glycans on bri1-5, while overexpression of either AtEDEM1 or AtEDEM2 stimulates bri1-5 ERAD and enhances the bri1-5 dwarfism. We concluded that, despite its functional redundancy with AtEDEM2, AtEDEM1 plays a predominant role in promoting bri1-5 degradation.

PMID: 35874007


Front Plant Sci , IF:5.753 , 2022 , V13 : P834027 doi: 10.3389/fpls.2022.834027

Identification and Characterization of Long Non-coding RNA in Tomato Roots Under Salt Stress.

Li, Ning and Wang, Zhongyu and Wang, Baike and Wang, Juan and Xu, Ruiqiang and Yang, Tao and Huang, Shaoyong and Wang, Huan and Yu, Qinghui

Institute of Horticulture Crops, Xinjiang Academy of Agricultural Sciences, Urumqi, China.; Key Laboratory of Horticulture Crop Genomics and Genetic Improvement in Xinjiang, Urumqi, China.; Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing, China.

As one of the most important vegetable crops in the world, the production of tomatoes was restricted by salt stress. Therefore, it is of great interest to analyze the salt stress tolerance genes. As the non-coding RNAs (ncRNAs) with a length of more than 200 nucleotides, long non-coding RNAs (lncRNAs) lack the ability of protein-coding, but they can play crucial roles in plant development and response to abiotic stresses by regulating gene expression. Nevertheless, there are few studies on the roles of salt-induced lncRNAs in tomatoes. Therefore, we selected wild tomato Solanum pennellii (S. pennellii) and cultivated tomato M82 to be materials. By high-throughput sequencing, 1,044 putative lncRNAs were identified here. Among them, 154 and 137 lncRNAs were differentially expressed in M82 and S. pennellii, respectively. Through functional analysis of target genes of differentially expressed lncRNAs (DE-lncRNAs), some genes were found to respond positively to salt stress by participating in abscisic acid (ABA) signaling pathway, brassinosteroid (BR) signaling pathway, ethylene (ETH) signaling pathway, and anti-oxidation process. We also construct a salt-induced lncRNA-mRNA co-expression network to dissect the putative mechanisms of high salt tolerance in S. pennellii. We analyze the function of salt-induced lncRNAs in tomato roots at the genome-wide levels for the first time. These results will contribute to understanding the molecular mechanisms of salt tolerance in tomatoes from the perspective of lncRNAs.

PMID: 35865296


Theor Appl Genet , IF:5.699 , 2022 Aug , V135 (8) : P2907-2923 doi: 10.1007/s00122-022-04158-0

The brassinosteroid biosynthesis gene TaD11-2A controls grain size and its elite haplotype improves wheat grain yields.

Xu, Huiyuan and Sun, Han and Dong, Jiajin and Ma, Chengxue and Li, Jingxue and Li, Zhuochun and Wang, Yihuan and Ji, Junqi and Hu, Xinrong and Wu, Meihui and Zhao, Chunhua and Qin, Ran and Wu, Jiajie and Ni, Fei and Cui, Fa and Wu, Yongzhen

Key Laboratory of Molecular Module-Based Breeding of High Yield and Abiotic Resistant Plants in Universities of Shandong, College of Agriculture, Ludong University, Yantai, Shandong, China.; State Key Laboratory of Crop Biology, College of Agronomy, Shandong Agricultural University, Taian, Shandong, China.; Key Laboratory of Molecular Module-Based Breeding of High Yield and Abiotic Resistant Plants in Universities of Shandong, College of Agriculture, Ludong University, Yantai, Shandong, China. sdaucf@126.com.; Key Laboratory of Molecular Module-Based Breeding of High Yield and Abiotic Resistant Plants in Universities of Shandong, College of Agriculture, Ludong University, Yantai, Shandong, China. yongzhenwu1204@163.com.

KEY MESSAGE: TaD11-2A affects grain size and root length and its natural variations are associated with significant differences in yield-related traits in wheat. Brassinosteroids (BRs) control many important agronomic traits and therefore the manipulation of BR components could improve crop productivity and performance. However, the potential effects of BR-related genes on yield-related traits and stress tolerance in wheat (Triticum aestivum L.) remain poorly understood. Here, we identified TaD11 genes in wheat (rice D11 orthologs) that encoded enzymes involved in BR biosynthesis. TaD11 genes were highly expressed in roots (Zadoks scale: Z11) and grains (Z75), while expression was significantly suppressed by exogenous BR (24-epiBL). Ectopic expression of TaD11-2A rescued the abnormal panicle structure and plant height (PH) of the clustered primary branch 1 (cpb1) mutant, and also increased endogenous BR levels, resulting in improved grain yields and grain quality in rice. The tad11-2a-1 mutant displayed dwarfism, smaller grains, sensitivity to 24-epiBL, and reduced endogenous BR contents. Natural variations in TaD11-2A were associated with significant differences in yield-related traits, including PH, grain width, 1000-grain weight, and grain yield per plant, and its favorable haplotype, TaD11-2A-HapI was subjected to positive selection during wheat breeding. Additionally, TaD11-2A influenced root length and salt tolerance in rice and wheat at seedling stages. These results indicated the important role of BR TaD11 biosynthetic genes in controlling grain size and root length, and also highlighted their potential in the molecular biological analysis of wheat.

PMID: 35794218


Plant Sci , IF:4.729 , 2022 Jul , V320 : P111281 doi: 10.1016/j.plantsci.2022.111281

Enhanced brassinosteroid signaling via the overexpression of SlBRI1 positively regulates the chilling stress tolerance of tomato.

Wang, Dan and Yang, Zaijun and Wu, Meiqi and Wang, Wei and Wang, Yue and Nie, Shuming

Key Laboratory of Southwest China Wildlife Resources Conservation (Ministry of Education), College of Life Science, China West Normal University, Nanchong 637009, China.; Key Laboratory of Southwest China Wildlife Resources Conservation (Ministry of Education), College of Life Science, China West Normal University, Nanchong 637009, China. Electronic address: nieshuming@163.com.

Brassinosteroids (BRs) regulate plant development and response to stress. BRASSINOSTEROID INSENSITIVE 1 (BRI1) is a BR receptor that activates BR signaling. Although the function of the tomato BR receptor SlBRI1 in regulating growth and drought resistance has been examined, that of SlBRI1 in cold tolerance is unclear. This study indicated that the expression of SlBRI1 in tomato was rapidly induced and reached its highest level at 3 h under chilling treatment and then decreased. The overexpression of SlBRI1 displayed low relative electrolyte leakage, malondialdehyde content, and reactive oxygen species (ROS) accumulation under chilling stress. The proline content and activities of superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) in SlBRI1OE plants were higher than those in the wild-type (WT) plants after chilling stress. The transcript abundances of five cold-responsive genes were higher in SlBRI1OE plants than in WT plants during chilling stress. RNA sequence analysis showed that the expression of the majority of genes encoding photosystem I and II were downregulated. The degree of suppression in SlBRI1OE plants was weaker than that in WT plants. Additionally, the Pn and Fv/Fm of SlBRI1OE plants were significantly higher than those of WT plants under chilling stress. We identified several genes encoding key enzymes in BRs; indole acetic acid (IAA), gibberellin (GA), and abscisic acid (ABA) biosynthesis or signaling were upregulated or downregulated during chilling stress. Chilling stress decreased the BRs and GA3 content, and increased IAA and ABA content. The contents were lower or higher in SlBRI1OE than in WT plants. Furthermore, chilling stress regulated the expression levels of 43 transcription factors. The expression of seven cold-regulated protein genes was higher or lower in SlBRI1OE plants than in WT plants under chilling stress. These results suggest that SlBRI1 positively regulates chilling tolerance mainly through ICE1-CBF-COR pathway in tomato.

PMID: 35643607


Front Genet , IF:4.599 , 2022 , V13 : P953458 doi: 10.3389/fgene.2022.953458

In-Silico Study of Brassinosteroid Signaling Genes in Rice Provides Insight Into Mechanisms Which Regulate Their Expression.

Ahmar, Sunny and Gruszka, Damian

Institute of Biology, Biotechnology and Environmental Protection, Faculty of Natural Sciences, University of Silesia, Katowice, Poland.

Brassinosteroids (BRs) regulate a diverse spectrum of processes during plant growth and development and modulate plant physiology in response to environmental fluctuations and stress factors. Thus, the BR signaling regulators have the potential to be targeted for gene editing to optimize the architecture of plants and make them more resilient to environmental stress. Our understanding of the BR signaling mechanism in monocot crop species is limited compared to our knowledge of this process accumulated in the model dicot species - Arabidopsis thaliana. A deeper understanding of the BR signaling and response during plant growth and adaptation to continually changing environmental conditions will provide insight into mechanisms that govern the coordinated expression of the BR signaling genes in rice (Oryza sativa) which is a model for cereal crops. Therefore, in this study a comprehensive and detailed in silico analysis of promoter sequences of rice BR signaling genes was performed. Moreover, expression profiles of these genes during various developmental stages and reactions to several stress conditions were analyzed. Additionally, a model of interactions between the encoded proteins was also established. The obtained results revealed that promoters of the 39 BR signaling genes are involved in various regulatory mechanisms and interdependent processes that influence growth, development, and stress response in rice. Different transcription factor-binding sites and cis-regulatory elements in the gene promoters were identified which are involved in regulation of the genes' expression during plant development and reactions to stress conditions. The in-silico analysis of BR signaling genes in O. sativa provides information about mechanisms which regulate the coordinated expression of these genes during rice development and in response to other phytohormones and environmental factors. Since rice is both an important crop and the model species for other cereals, this information may be important for understanding the regulatory mechanisms that modulate the BR signaling in monocot species. It can also provide new ways for the plant genetic engineering technology by providing novel potential targets, either cis-elements or transcriptional factors, to create elite genotypes with desirable traits.

PMID: 35873468


Front Genet , IF:4.599 , 2022 , V13 : P891702 doi: 10.3389/fgene.2022.891702

RNA-Seq and Gene Ontology Analysis Reveal Differences Associated With Low R/FR-Induced Shade Responses in Cultivated Lentil and a Wild Relative.

Yuan, Hai Ying and Caron, Carolyn T and Vandenberg, Albert and Bett, Kirstin E

Department of Plant Sciences, University of Saskatchewan, Saskatoon, SK, Canada.; Aquatic and Crop Resource Development Research Center, National Research Council of Canada, Saskatoon, SK, Canada.

Lentil is an important pulse crop not only because of its high nutrient value but also because of its ecological advantage in a sustainable agricultural system. Our previous work showed that the cultivated lentil and wild lentil germplasm respond differently to light environments, especially to low R/FR-induced shade conditions. Little is known about how cultivated and wild lentils respond to shade at the level of gene expression and function. In this study, transcriptomic profiling of a cultivated lentil (Lupa, L. culinaris) and a wild lentil (BGE 016880, L. orientalis) at several growth stages is presented. De novo transcriptomes were assembled for both genotypes, and differential gene expression analysis and gene ontology enrichment analysis were performed. The transcriptomic resources generated in this study provide fundamental information regarding biological processes and genes associated with shade responses in lentils. BGE 016880 and Lupa shared a high similarity in their transcriptomes; however, differential gene expression profiles were not consistent between these two genotypes. The wild lentil BGE 016880 had more differentially expressed genes than the cultivated lentil Lupa. Upregulation of genes involved in gibberellin, brassinosteroid, and auxin synthesis and signaling pathways, as well as cell wall modification, in both genotypes explains their similarity in stem elongation response under the shade. Genes involved in jasmonic acid and flavonoid biosynthesis pathways were downregulated in BGE 016880 only, and biological processes involved in defense responses were significantly enriched in the wild lentil BGE 016880 only. Downregulation of WRKY and MYB transcription factors could contribute to the reduced defense response in BGE 016880 but not in Lupa under shade conditions. A better understanding of shade responses of pulse crop species and their wild relatives will play an important role in developing genetic strategies for crop improvement in response to changes in light environments.

PMID: 35795209


Sci Rep , IF:4.379 , 2022 Jul , V12 (1) : P11294 doi: 10.1038/s41598-022-15284-6

A brassinosteroid functional analogue increases soybean drought resilience.

Perez-Borroto, Lucia Sandra and Guzzo, Maria Carla and Posada, Gisella and Pena Malavera, Andrea Natalia and Castagnaro, Atilio Pedro and Gonzalez-Olmedo, Justo Lorenzo and Coll-Garcia, Yamilet and Pardo, Esteban Mariano

Plant Breeding, Wageningen University and Research, 6708 PB, Wageningen, The Netherlands.; Instituto de Fisiologia y Recursos Geneticos Vegetales Victorio S. Trippi - Unidad de Estudios Agropecuarios (IFRGV-UDEA, INTA-CONICET), Av. 11 de septiembre 4755, CP X5014MGO, Cordoba, Argentina.; Instituto de Tecnologia Agroindustrial del Noroeste Argentino (ITANOA), Estacion Experimental Agroindustrial Obispo Colombres (EEAOC) /Consejo Nacional de Investigaciones Cientificas Y Tecnicas (CONICET), Las Talitas, Tucuman, Argentina.; Centro de Bioplantas, Universidad de Ciego de Avila "Maximo Gomez Baez", Ciego de Avila, Cuba.; Centro de Estudios de Productos Naturales, Facultad de Quimica, Universidad de La Habana, Havana, Cuba.; Instituto de Tecnologia Agroindustrial del Noroeste Argentino (ITANOA), Estacion Experimental Agroindustrial Obispo Colombres (EEAOC) /Consejo Nacional de Investigaciones Cientificas Y Tecnicas (CONICET), Las Talitas, Tucuman, Argentina. empardokarate@gmail.com.

Drought severely affects soybean productivity, challenging breeding/management strategies to increase crop resilience. Hormone-based biostimulants like brassinosteroids (BRs) modulate growth/defence trade-off, mitigating yield losses; yet, natural molecule's low stability challenges the development of cost-effective and long-lasting analogues. Here, we investigated for the first time the effects of BR functional analogue DI-31 in soybean physiology under drought by assessing changes in growth, photosynthesis, water relations, antioxidant metabolism, nodulation, and nitrogen homeostasis. Moreover, DI-31 application frequencies' effects on crop cycle and commercial cultivar yield stabilisation under drought were assessed. A single foliar application of DI-31 favoured plant drought tolerance, preventing reductions in canopy development and enhancing plant performance and water use since the early stages of stress. The analogue also increased the antioxidant response, favouring nitrogen homeostasis maintenance and attenuating the nodular senescence. Moreover, foliar applications of DI-31 every 21 days enhanced the absolute yield by ~ 9% and reduced drought-induced yield losses by ~ 7% in four commercial cultivars, increasing their drought tolerance efficiency by ~ 12%. These findings demonstrated the practical value of DI-31 as an environmentally friendly alternative for integrative soybean resilience management under drought.

PMID: 35788151


Plant Physiol Biochem , IF:4.27 , 2022 Aug , V185 : P325-335 doi: 10.1016/j.plaphy.2022.06.011

ZmBSK1 positively regulates BR-induced H2O2 production via NADPH oxidase and functions in oxidative stress tolerance in maize.

Liu, Lei and Sun, Yanchao and Zhang, Meijing and Liu, Ruixiang and Wu, Xiaming and Chen, Yanping and Yuan, Jianhua

Provincial Key Laboratory of Agrobiology, Institute of Food Crops, Jiangsu Academy of Agricultural Sciences, Nanjing, 210014, Jiangsu, China.; Provincial Key Laboratory of Agrobiology, Institute of Food Crops, Jiangsu Academy of Agricultural Sciences, Nanjing, 210014, Jiangsu, China; College of Life Sciences, Nanjing Agricultural University, Nanjing, 210095, Jiangsu, China.; Provincial Key Laboratory of Agrobiology, Institute of Food Crops, Jiangsu Academy of Agricultural Sciences, Nanjing, 210014, Jiangsu, China. Electronic address: chenyp@jaas.ac.cn.; Provincial Key Laboratory of Agrobiology, Institute of Food Crops, Jiangsu Academy of Agricultural Sciences, Nanjing, 210014, Jiangsu, China. Electronic address: yuanjh1123@163.com.

Brassinosteroid (BR) has been indicated to induce the production of hydrogen peroxide (H2O2) in plants in response to various environmental stimuli. However, it remains largely unknown how BR induces H2O2 production. In this study, we found that BR treatment significantly raised the kinase activity of maize (Zea mays L.) brassinosteroid-signaling kinase 1 (ZmBSK1) using the immunoprecipitation kinase assay. ZmBSK1 could modulate the gene expressions and activities of nicotinamide adenine dinucleotide phosphate (NADPH) oxidases (EC 1.6.3.1) to modulate BR-induced H2O2 production. BR could enhance the interaction between ZmBSK1 and maize calcium/calmodulin-dependent protein kinase (ZmCCaMK), a previously identified substrate of ZmBSK1. The BR-induced phosphorylation and kinase activity of ZmCCaMK are dependent on ZmBSK1. Moreover, we showed that ZmBSK1 regulated the NADPH oxidase gene expression and activity via directly phosphorylating ZmCCaMK. Genetic analysis suggested that ZmBSK1-ZmCCaMK module strengthened plant tolerance to oxidative stress induced by exogenous application of H2O2 through improving the activities of antioxidant defense enzyme and alleviating the malondialdehyde (MDA) accumulation and electrolyte leakage rate. In conclusion, these findings provide the new insights of ZmBSK1 functioning in BR-induced H2O2 production and the theoretical supports for breeding stress-tolerant crops.

PMID: 35738188


BMC Plant Biol , IF:4.215 , 2022 Jul , V22 (1) : P319 doi: 10.1186/s12870-022-03701-x

Integrated transcriptome and hormonal analysis of naphthalene acetic acid-induced adventitious root formation of tea cuttings (Camellia sinensis).

Wang, Yongxin and Pang, Dandan and Ruan, Li and Liang, Jinbo and Zhang, Qiang and Qian, Yinhong and Zhang, Yazhen and Bai, Peixian and Wu, Liyun and Cheng, Hao and Cui, Qingmei and Wang, Liyuan and Wei, Kang

Key Laboratory of Tea Biology and Resources Utilization, Ministry of Agriculture, National Center for Tea Improvement, Tea Research Institute Chinese Academy of Agricultural Sciences (TRICAAS), Hangzhou, 310008, China.; Tea Research Institute, Yunnan Academy of Agricultural Sciences, Menghai, 666201, China.; Tea Research Institute of Enshi Academy of Agricultural Sciences, Enshi, 445000, China.; Tea Research Institute of Enshi Academy of Agricultural Sciences, Enshi, 445000, China. hbeshsz@yahoo.com.cn.; Key Laboratory of Tea Biology and Resources Utilization, Ministry of Agriculture, National Center for Tea Improvement, Tea Research Institute Chinese Academy of Agricultural Sciences (TRICAAS), Hangzhou, 310008, China. wangly@tricaas.com.; Key Laboratory of Tea Biology and Resources Utilization, Ministry of Agriculture, National Center for Tea Improvement, Tea Research Institute Chinese Academy of Agricultural Sciences (TRICAAS), Hangzhou, 310008, China. weikang@tricaas.com.

BACKGROUND: Tea plant breeding or cultivation mainly involves propagation via cuttings, which not only ensures the inheritance of the excellent characteristics of the mother plant but also facilitates mechanized management. The formation of adventitious root (AR) determines the success of cutting-based propagation, and auxin is an essential factor involved in this process. To understand the molecular mechanism underlying AR formation in nodal tea cuttings, transcriptome and endogenous hormone analysis was performed on the stem bases of red (mature)- and green (immature)-stem cuttings of 'Echa 1 hao' tea plant as affected by a pulse treatment with naphthalene acetic acid (NAA). RESULTS: In this study, NAA significantly promoted AR formation in both red- and green-stem cuttings but slightly reduced callus formation. External application of NAA reduced the levels of endogenous indole-3-acetic acid (IAA) and cytokinin (TZR, trans-zeatin riboside). The number of DEGs (NAA vs. CK) identified in the green-stem cuttings was significantly higher than that in the red-stem cuttings, which corresponded to a higher rooting rate of green-stem cuttings under the NAA treatment. A total of 82 common DEGs were identified as being hormone-related and involved in the auxin, cytokinin, abscisic acid, ethylene, salicylic acid, brassinosteroid, and jasmonic acid pathways. The negative regulation of NAA-induced IAA and GH3 genes may explain the decrease of endogenous IAA. NAA reduced endogenous cytokinin levels and further downregulated the expression of cytokinin signalling-related genes. By the use of weighted gene co-expression network analysis (WGCNA), several hub genes, including three [cellulose synthase (CSLD2), SHAVEN3-like 1 (SVL1), SMALL AUXIN UP RNA (SAUR21)] that are highly related to root development in other crops, were identified that might play important roles in AR formation in tea cuttings. CONCLUSIONS: NAA promotes the formation of AR of tea cuttings in coordination with endogenous hormones. The most important endogenous AR inductor, IAA, was reduced in response to NAA. DEGs potentially involved in NAA-mediated AR formation of tea plant stem cuttings were identified via comparative transcriptome analysis. Several hub genes, such as CSLD2, SVL1 and SAUR21, were identified that might play important roles in AR formation in tea cuttings.

PMID: 35787241


Planta , IF:4.116 , 2022 Jul , V256 (2) : P27 doi: 10.1007/s00425-022-03939-7

Natural variation of the BRD2 allele affects plant height and grain size in rice.

Huang, Jinpeng and Chen, Zhiming and Lin, Jiajia and Chen, Jinwen and Wei, Menghao and Liu, Liang and Yu, Feng and Zhang, Zesen and Chen, Fangyu and Jiang, Liangrong and Zheng, Jingsheng and Wang, Tiansheng and Chen, Huiqing and Xie, Wangyou and Huang, Senhao and Wang, Houcong and Huang, Yumin and Huang, Rongyu

School of Life Sciences, Xiamen University, Xiamen, China.; Quanzhou Agricultural Science Institute, Quanzhou, 362212, China.; Key Laboratory of Ministry of Education for Genetic Improvement and Comprehensive Utilization of Crops, Fujian Provincial Key Laboratory of Crop Breeding by Design, College of Agriculture, Fujian Agriculture and Forestry University, Fuzhou, China.; School of Life Sciences, Xiamen University, Xiamen, China. huangrongyu@xmu.edu.cn.

MAIN CONCLUSION: The zqdm1 identified from a rice mutant is a novel allele of BRD2 and is responsible for regulating rice plant height, grain size and appearance, which has possibilities on improving rice quality. Plant height is an important agronomic trait related to rice yield, and grain size directly determines grain yield in rice (Oryza sativa L.). With the development of molecular biotechnology and genome sequencing technology, more and more key genes associated with plant height and grain size have been cloned and identified in recent years. This study identified the zqdm1 gene from a mutant with reduced plant height and grain size. The zqdm1 gene was revealed to be a new allele of BRASSINOSTEROID DEFICIENT DWARF 2 (BRD2), encoding a FAD-linked oxidoreductase protein involved in the brassinosteroid (BR) biosynthesis pathway, and regulates plant height by reducing cell number of longitudinal sections of the internode and regulates grain size by altering cell expansion. A 369-bp DNA fragment was found inserted at the first exon, resulting in protein-coding termination. This mutation has not been discovered in previous studies. Complementation tests have confirmed that 369-bp insertion in BRD2 was responsible for the plant height and grain size changing in the zqdm1 mutant. Over-expression of BRD2 driven by different promoters into indica rice variety Jiafuzhan (JFZ) results in slender grains, suggesting its function on regulating grain shape. In summary, the current study has identified a new BRD2 allele, which facilitated the further research on the molecular mechanism of this gene on regulating growth and development.

PMID: 35780402


Membranes (Basel) , IF:4.106 , 2022 Jun , V12 (6) doi: 10.3390/membranes12060606

Plasma Membrane-Associated Proteins Identified in Arabidopsis Wild Type, lbr2-2 and bak1-4 Mutants Treated with LPSs from Pseudomonas syringae and Xanthomonas campestris.

Offor, Benedict C and Mhlongo, Msizi I and Dubery, Ian A and Piater, Lizelle A

Department of Biochemistry, University of Johannesburg, Auckland Park, Johannesburg 2006, South Africa.

Plants recognise bacterial microbe-associated molecular patterns (MAMPs) from the environment via plasma membrane (PM)-localised pattern recognition receptor(s) (PRRs). Lipopolysaccharides (LPSs) are known as MAMPs from gram-negative bacteria that are most likely recognised by PRRs and trigger defence responses in plants. The Arabidopsis PRR(s) and/or co-receptor(s) complex for LPS and the associated defence signalling remains elusive. As such, proteomic identification of LPS receptors and/or co-receptor complexes will help to elucidate the molecular mechanisms that underly LPS perception and defence signalling in plants. The Arabidopsis LPS-binding protein (LBP) and bactericidal/permeability-increasing protein (BPI)-related-2 (LBR2) have been shown to recognise LPS and trigger defence responses while brassinosteroid insensitive 1 (BRI1)-associated receptor kinase 1 (BAK1) acts as a co-receptor for several PRRs. In this study, Arabidopsis wild type (WT) and T-DNA knock out mutants (lbr2-2 and bak1-4) were treated with LPS chemotypes from Pseudomonas syringae pv. tomato DC3000 (Pst) and Xanthomonas campestris pv. campestris 8004 (Xcc) over a 24 h period. The PM-associated protein fractions were separated by liquid chromatography and analysed by tandem mass spectrometry (LC-MS/MS) followed by data analysis using Byonic(TM) software. Using Gene Ontology (GO) for molecular function and biological processes, significant LPS-responsive proteins were grouped according to defence and stress response, perception and signalling, membrane transport and trafficking, metabolic processes and others. Venn diagrams demarcated the MAMP-responsive proteins that were common and distinct to the WT and mutant lines following treatment with the two LPS chemotypes, suggesting contributions from differential LPS sub-structural moieties and involvement of LBR2 and BAK1 in the LPS-induced MAMP-triggered immunity (MTI). Moreover, the identification of RLKs and RLPs that participate in other bacterial and fungal MAMP signalling proposes the involvement of more than one receptor and/or co-receptor for LPS perception as well as signalling in Arabidopsis defence responses.

PMID: 35736313


BMC Genomics , IF:3.969 , 2022 Jun , V23 (1) : P453 doi: 10.1186/s12864-022-08684-5

Genome-wide identification and expression analysis reveals spinach brassinosteroid-signaling kinase (BSK) gene family functions in temperature stress response.

Li, Yang and Zhang, Heng and Zhang, Yongxue and Liu, Yanshuang and Li, Yueyue and Tian, Haodong and Guo, Siyi and Sun, Meihong and Qin, Zhi and Dai, Shaojun

Development Center of Plant Germplasm Resources, College of Life Sciences, Shanghai Normal University, Shanghai, 200234, China.; Development Center of Plant Germplasm Resources, College of Life Sciences, Shanghai Normal University, Shanghai, 200234, China. zhangheng1029@163.com.; Key Laboratory of Saline-alkali Vegetation Ecology Restoration (Northeast Forestry University), Ministry of Education, College of Life Sciences, Northeast Forestry University, Harbin, 150040, China.; State Key Laboratory of Crop Stress Adaptation and Improvement, School of Life Sciences, Henan University, Kaifeng, 475004, China.; Development Center of Plant Germplasm Resources, College of Life Sciences, Shanghai Normal University, Shanghai, 200234, China. daishaojun@shnu.edu.cn.

BACKGROUND: Brassinosteroid (BR)- signaling kinase (BSK) is a critical family of receptor-like cytoplasmic kinase for BR signal transduction, which plays important roles in plant development, immunity, and abiotic stress responses. Spinach (Spinacia oleracea) is cold- tolerant but heat- sensitive green leafy vegetable. A study on BSK family members and BSKs- mediated metabolic processes in spinach has not been performed. RESULTS: We identified and cloned seven SoBSKs in spinach. Phylogenetic and collinearity analyses suggested that SoBSKs had close relationship with dicotyledonous sugar beet (Beta vulgaris) rather than monocotyledons. The analyses of gene structure and conserved protein domain/ motif indicated that most SoBSKs were relative conserved, while SoBSK6 could be a truncated member. The prediction of post-translation modification (PTM) sites in SoBSKs implied their possible roles in signal transduction, redox regulation, and protein turnover of SoBSKs, especially the N-terminal myristoylation site was critical for BSK localization to cell periphery. Cis-acting elements for their responses to light, drought, temperature (heat and cold), and hormone distributed widely in the promoters of SoBSKs, implying the pivotal roles of SoBSKs in response to diverse abiotic stresses and phytohormone stimuli. Most SoBSKs were highly expressed in leaves, except for SoBSK7 in roots. Many SoBSKs were differentially regulated in spinach heat- sensitive variety Sp73 and heat- tolerant variety Sp75 under the treatments of heat, cold, as well as exogenous brassinolide (BL) and abscisic acid (ABA). The bsk134678 mutant Arabidopsis seedlings exhibited more heat tolerance than wild- type and SoBSK1- overexpressed seedlings. CONCLUSIONS: A comprehensive genome- wide analysis of the BSK gene family in spinach presented a global identification and functional prediction of SoBSKs. Seven SoBSKs had relatively- conserved gene structure and protein function domains. Except for SoBSK6, all the other SoBSKs had similar motifs and conserved PTM sites. Most SoBSKs participated in the responses to heat, cold, BR, and ABA. These findings paved the way for further functional analysis on BSK- mediated regulatory mechanisms in spinach development and stress response.

PMID: 35725364


Plants (Basel) , IF:3.935 , 2022 Jun , V11 (13) doi: 10.3390/plants11131728

HD-Zip III Gene Family: Identification and Expression Profiles during Leaf Vein Development in Soybean.

Gao, Jing and Chen, Jiyu and Feng, Lingyang and Wang, Qi and Li, Shenglan and Tan, Xianming and Yang, Feng and Yang, Wenyu

College of Agronomy, Sichuan Agricultural University, Huimin Road 211, Wenjiang District, Chengdu 611130, China.; Sichuan Engineering Research Center for Crop Strip Intercropping System, Chengdu 611130, China.; Key Laboratory of Crop Ecophysiology and Farming System in Southwest, Ministry of Agriculture, Chengdu 611130, China.

Leaf veins constitute the transport network for water and photosynthetic assimilates in vascular plants. The class III homeodomain-leucine zipper (HD-Zip III) gene family is central to the regulation of vascular development. In this research, we performed an overall analysis of the HD-Zip III genes in soybean (Glycine max L. Merr.). Our analysis included the phylogeny, conservation domains and cis-elements in the promoters of these genes. We used the quantitative reverse transcription-polymerase chain reaction to characterize the expression patterns of HD-Zip III genes in leaf vein development and analyze the effects of exogenous hormone treatments. In this study, twelve HD-Zip III genes were identified from the soybean genome and named. All soybean HD-Zip III proteins contained four highly conserved domains. GmHB15-L-1 transcripts showed steadily increasing accumulation during all stages of leaf vein development and were highly expressed in cambium cells. GmREV-L-1 and GmHB14-L-2 had nearly identical expression patterns in soybean leaf vein tissues. GmREV-L-1 and GmHB14-L-2 transcripts remained at stable high levels at all xylem developmental stages. GmREV-L-1 and GmHB14-L-2 were expressed at high levels in the vascular cambium and xylem cells. Overall, GmHB15-L-1 may be an essential regulator that is responsible for the formation or maintenance of soybean vein cambial cells. GmREV-L-1 and GmHB14-L-2 were correlated with xylem differentiation in soybean leaf veins. This study will pave the way for identifying the molecular mechanism of leaf vein development.

PMID: 35807680


Biomed Res Int , IF:3.411 , 2022 , V2022 : P8445484 doi: 10.1155/2022/8445484

Transcriptomic Insight into Viviparous Growth in Water Lily.

Su, Qun and Wang, Hong-Yan and Tian, Min and Li, Chun-Niu and Li, Xian-Min and Huang, Zhan-Wen and Bu, Zhao-Yang and Lu, Jia-Shi

Flower Research Institute of Guangxi Academy of Agricultural Sciences, Nanning Guangxi 530007, China.; Flower Research Institute of Yunnan Academy of Agricultural Sciences, Kunming Yunnan 650200, China.; National Engineering Research Center for Ornamental Horticulture, Kunming Yunnan 650200, China.

Water lily is an important ornamental flower plant which is capable of viviparous plantlet development. But no study has been reported on the molecular basis of viviparity in water lily. Hence, we performed a comparative transcriptome study between viviparous water lily Nymphaea micrantha and a nonviviparous species Nymphaea colorata at four developmental stages. The higher expression of highly conserved AUX/IAA, ARF, GH3, and SAUR gene families in N. micrantha compared to N. colorata is predicted to have a major impact on the development and evolution of viviparity in water lily. Likewise, differential regulation of hormone signaling, brassinosteroid, photosynthesis, and energy-related pathways in the two species provide clues of their involvement in viviparity phenomenon. This study revealed the complex mechanism of viviparity trait in water lily. The transcriptomic signatures identified are important basis for future breeding and research of viviparity in water lily and other plant species.

PMID: 35845943


Biosci Biotechnol Biochem , IF:2.043 , 2022 Jul , V86 (8) : P1004-1012 doi: 10.1093/bbb/zbac074

Structure modification of nonsteroidal brassinolide-like compound, NSBR1.

Takimoto, Seisuke and Nishikawa, Bunta and Matsuo, Midori and Hinata, Shiori and Hisatomi, Taiki and Yamagami, Ayumi and Nakano, Takeshi and Nakagawa, Yoshiaki and Miyagawa, Hisashi

Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kyoto, Japan.; Central Pharmaceutical Research Institute, Japan Tobacco Inc., 1-1 Murasaki-cho, Takatsuki, Osaka, Japan.; Graduate School of Biostudies, Kyoto University, Kyoto, Japan.

Brassinolide (BL) is a possible plant growth regulator in agriculture, but the presence of a steroid skeleton hampers the field application of BL in agriculture because of its high synthetic cost. We discovered NSBR1 as the first nonsteroidal BL-like compound using in silico technology. Searching for more potent BL-like compounds, we modified the structure of NSBR1 with respect to 2 benzene rings and the piperazine ring. The activity of synthesized compounds was measured in Arabidopsis hypocotyl elongation. The propyl group of butyryl moiety of NSBR1 was changed to various alkyl groups, such as straight, branched, and cyclic alkyl chains. Another substituent, F, at the ortho position of the B ring toward the piperazine ring was changed to other substituents. A methyl group was introduced to the piperazine ring. Most of the newly synthesized compounds with the 3,4-(OH)2 group at the A ring significantly elongated the hypocotyl of Arabidopsis.

PMID: 35687006


Biosci Biotechnol Biochem , IF:2.043 , 2022 Jul , V86 (8) : P1041-1048 doi: 10.1093/bbb/zbac071

BRZ-INSENSITIVE-PALE GREEN 1 is encoded by chlorophyll biosynthesis enzyme gene that functions in the downstream of brassinosteroid signaling.

Tachibana, Ryo and Yamagami, Ayumi and Miyagi, Shino and Nakazawa-Miklasevica, Miki and Matsui, Minami and Sakuta, Masaaki and Tanaka, Ryouichi and Asami, Tadao and Nakano, Takeshi

Graduate School of Biostudies, Kyoto University, KItashirakawa-Oiwake-cho, Sakyo-ku, Kyoto, Japan.; Department of Biology, Ochanomizu University, Bunkyo-ku, Tokyo, Japan.; RIKEN, CSRS, Tsurumi-ku, Yokohama, Japan.; The Institute of Oncology, Riga Stradins University, 16 Dzirciema Street, Riga, Latvia.; Organization for the Strategic Coordination of Research and Intellectual Properties, Meiji University, Tama-ku, Kawasaki, Japan.; Institute of Low Temperature Science, Hokkaido University, Kita-ku, Sapporo, Japan.; Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Bunkyo-ku, Tokyo, Japan.

Brassinosteroids (BRs), a kind of phytohormone, have various biological activities such as promoting plant growth, increasing stress resistance, and chloroplast development. Though BRs have been known to have physiological effects on chloroplast, the detailed mechanism of chloroplast development and chlorophyll biosynthesis in BR signaling remains unknown. Here we identified a recessive pale green Arabidopsis mutant, Brz-insensitive-pale green1 (bpg1), which was insensitive to promoting of greening by BR biosynthesis-specific inhibitor Brz in the light. BPG1 gene encoded chlorophyll biosynthesis enzyme, 3, 8-divinyl protochlorophyllide a 8-vinyl reductase (DVR), and bpg1 accumulated divinyl chlorophylls. Chloroplast development was suppressed in bpg1. Brz dramatically increased the expression of chlorophyll biosynthesis enzyme genes, including BPG1. These results suggest that chlorophyll biosynthesis enzymes are regulated by BR signaling in the aspect of gene expression and BPG1 plays an important role in regulating chloroplast development.

PMID: 35583242


Genes Genomics , IF:1.839 , 2022 Jul , V44 (7) : P833-841 doi: 10.1007/s13258-022-01266-5

Role of tyrosine autophosphorylation and methionine residues in BRI1 function in Arabidopsis thaliana.

Choi, Jae-Han and Oh, Eun-Seok and Min, Hansol and Chae, Won Byoung and Mandadi, Kranthi Kiran and Oh, Man-Ho

Department of Biological Sciences, Chungnam National University, Daejeon, 34134, Korea.; Department of Environmental Horticulture, Dankook University, Cheonan, 31116, Korea.; Department of Plant Pathology and Microbiology, Texas A&M University, College Station, TX, USA.; Department of Biological Sciences, Chungnam National University, Daejeon, 34134, Korea. manhooh@cnu.ac.kr.

BACKGROUND: Brassinosteroids (BRs), a group of plant growth hormones, control biomass accumulation and biotic and abiotic stress tolerance, and therefore are highly relevant to agriculture. BRs bind to the BR receptor protein, brassinosteroid insensitive 1 (BRI1), which is classified as a serine/threonine (Ser/Thr) protein kinase. Recently, we reported that BRI1 acts as a dual-specificity kinase both in vitro and in vivo by undergoing autophosphorylation at tyrosine (Tyr) residues. OBJECTIVE: In this study, we characterized the increased leaf growth and early flowering phenotypes of transgenic lines expressing the mutated recombinant protein, BRI1(Y831F)-Flag, compared with those expressing BRI1-Flag. BRI1(Y831F)-Flag transgenic plants showed a reduction in hypocotyl and petiole length compared with BRI1-Flag seedlings. Transcriptome analysis revealed differential expression of flowering time-associated genes (AP1, AP2, AG, FLC, and SMZ) between BRI1(Y831F)-Flag and BRI1-Flag transgenic seedlings. We also performed site-directed mutagenesis of the BRI1 gene, and investigated the effect of methionine (Met) substitution in the extracellular domain (ECD) of BRI1 on plant growth and BR sensitivity by evaluating hypocotyl elongation and root growth inhibition. METHODS: The pBIB-Hyg(+)-pBR-BRI1-Flag construct(Li et al. 2002) was used as the template for SDM with QuickChange XL Site Directed Mutagenesis Kit (Stratagene, La Jolla, CA, USA) to make the SDM mutants. After PCR with SDM kit, add 1 mul of Dpn1 to PCR reaction. Incubate at 37 degrees C for 2 h to digest parental DNA and then transformed into XL10-gold competent cells. Transcriptome analysis was carried out at the University of Illinois (Urbana-Champaign, Illinois, USA). RNA was prepared and hybridized to the Affymetrix GeneChip Arabidopsis ATH1 Genome Array using the Gene Chip Express Kit (Ambion, Austin, TX, USA). RESULTS: Tyrosine 831 autophosphorylation of BRI1 regulates Arabidopsis flowering time, and mutation of methionine residues in the extracellular domain of BRI1 affects hypocotyl and root length. BRI1(M656Q)-Flag, BRI1(M657Q)-Flag, and BRI1(M661Q)-Flag seedlings were insensitive to the BL treatment and showed no inhibition of root elongation. However, BRI1(M665Q)-Flag and BRI1(M671Q)-Flag seedlings were sensitive to the BL treatment, and exhibited root elongation inhibition. the early flowering phenotype of BRI1(Y831F)-Flag transgenic plants is consistent with the expression levels of key flowering-related genes, including those promoting flowering (AP1, AP2, and AG) and repressing flowering (FLC and SMZ).

PMID: 35598220


Braz J Biol , 2022 , V82 : Pe260818 doi: 10.1590/1519-6984.260818

Mitigation of the effects of salt stress in cowpea bean through the exogenous aplication of brassinosteroid.

Sousa, D J P and Nogueira, G A S and Teixeira, K B S and Monteiro, G G T N and Brito, A E A and Nascimento, V R and Albuquerque, G D P and Oliveira, T J M and Souza, L C and Freitas, J M N and Oliveira Neto, C F and Okumura, R S

Universidade Federal Rural da Amazonia - UFRA, Instituto de Ciencias Agrarias, Laboratorio de Fisiologia Vegetal, Grupo de Estudos da Biodiversidade em Plantas Superiores, Belem, PA, Brasil.; Universidade Federal Rural da Amazonia - UFRA, Instituto de Ciencias Agrarias, Laboratorio de Fisiologia Vegetal, Grupo de Estudos da Biodiversidade em Plantas Superiores, Parauapebas, PA, Brasil.

Cowpea (Vigna unguiculata (L.) Walp.) is a legume widely cultivated by small, medium and large producers in several Brazilian regions. However, one of the concerns for the production of cowpea in Brazil in recent years is the low rainfall activity in these regions, which generates the accumulation of salts on the surface. The objective of this work was to evaluate the effects of salt stress on growth parameters and enzyme activity in cowpea plants at different concentrations of brassinosteroids. Experiment was developed in a greenhouse using a completely randomized experimental design in a 3 x 3 factorial scheme. The treatments consisted of three levels of brassinosteroids (0, 3 and 6 microM EBL) and three levels of salt stress (0, 50 and 100 mM NaCl). Growth factors (height, diameter and number of leaves) decreased in the saline condition. With the presence of brassinosteroid the height did not increase, but the number of leaves did, mainly in the saline dosage of 100 mM NaCl. In the variable membrane integrity, brassinosteroid was efficient in both salinity dosages, the same not happening with the relative water content, where the saline condition did not affect the amount of water in the vegetable, with the application of brassino it remained high, decreasing only at dosage 100 mM NaCl. The nitrate reductase enzyme was greatly affected in the root system even with the application of increasing doses of brassino. Therefore, brassinosteroids as a promoter of saline tolerance in cowpea seedlings was positive. The concentration of 3microM of EBL provided the most satisfactory effect in tolerating the deleterious effects of the saline condition. The same cannot be concluded for the concentration of 6microM of EBL that did not promote tolerance to some variables.

PMID: 35857948


Plant Commun , 2022 Jun : P100348 doi: 10.1016/j.xplc.2022.100348

Deubiquitination of BES1 by UBP12/UBP13 promotes brassinosteroid signaling and plant growth.

Park, Su-Hyun and Jeong, Jin Seo and Zhou, Yu and Binte Mustafa, Nur Fatimah and Chua, Nam-Hai

Temasek Life Sciences Laboratory, National University of Singapore, 1 Research Link, Singapore 117604, Singapore.; Temasek Life Sciences Laboratory, National University of Singapore, 1 Research Link, Singapore 117604, Singapore. Electronic address: chua@rockefeller.edu.

As a key transcription factor in the brassinosteroid (BR) signaling pathway, the activity and expression of BES1 (BRI1-EMS-SUPPRESSOR 1) are stringently regulated. BES1 degradation is mediated by ubiquitin-related 26S proteasomal and autophagy pathways, which attenuate and terminate BR signaling; however, the opposing deubiquitinases (DUBs) are still unknown. Here, we showed that the ubp12-2w/13-3 double mutant phenocopies the BR-deficient dwarf mutant, suggesting that the two DUBs UBP12/UBP13 antagonize ubiquitin-mediated degradation to stabilize BES1. These two DUBs can trim tetraubiquitin with K46 and K63 linkages in vitro. UBP12/BES1 and UBP13/BES1 complexes are localized in both cytosol and nuclei. UBP12/13 can deubiquitinate polyubiquitinated BES1 in vitro and in planta, and UBP12 interacts with and deubiquitinates both inactive, phosphorylated BES1 and active, dephosphorylated BES1 in vivo. UBP12 overexpression in BES1(OE) plants significantly enhances cell elongation in hypocotyls and petioles and increases the ratio of leaf length to width compared with BES1(OE) or UBP12(OE) plants. Hypocotyl elongation and etiolation result from elevated BES1 levels because BES1 degradation is retarded by UBP12 in darkness or in light with BR. Protein degradation inhibitor experiments show that the majority of BES1 can be degraded by either the proteasomal or the autophagy pathway, but a minor BES1 fraction remains pathway specific. In conclusion, UBP12/UBP13 deubiquitinate BES1 to stabilize the latter as a positive regulator for BR responses.

PMID: 35706355


Plant Commun , 2022 Jul , V3 (4) : P100317 doi: 10.1016/j.xplc.2022.100317

A reference-guided TILLING by amplicon-sequencing platform supports forward and reverse genetics in barley.

Jiang, Congcong and Lei, Miaomiao and Guo, Yu and Gao, Guangqi and Shi, Lijie and Jin, Yanlong and Cai, Yu and Himmelbach, Axel and Zhou, Shenghui and He, Qiang and Yao, Xuefeng and Kan, Jinhong and Haberer, Georg and Duan, Fengying and Li, Lihui and Liu, Jun and Zhang, Jing and Spannagl, Manuel and Liu, Chunming and Stein, Nils and Feng, Zongyun and Mascher, Martin and Yang, Ping

Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing, China.; Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing, China; College of Agronomy, Sichuan Agricultural University, Chengdu, China.; Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), Seeland, Germany.; Institute of Botany, Chinese Academy of Sciences, Beijing, China.; Plant Genome and Systems Biology (PGSB), Helmholtz Center Munich, German Research Center for Environmental Health, Neuherberg, Germany.; College of Agronomy, Sichuan Agricultural University, Chengdu, China.; Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), Seeland, Germany. Electronic address: mascher@ipk-gatersleben.de.; Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing, China. Electronic address: yangping@caas.cn.

Barley is a diploid species with a genome smaller than those of other members of the Triticeae tribe, making it an attractive model for genetic studies in Triticeae crops. The recent development of barley genomics has created a need for a high-throughput platform to identify genetically uniform mutants for gene function investigations. In this study, we report an ethyl methanesulfonate (EMS)-mutagenized population consisting of 8525 M3 lines in the barley landrace "Hatiexi" (HTX), which we complement with a high-quality de novo assembly of a reference genome for this genotype. The mutation rate within the population ranged from 1.51 to 4.09 mutations per megabase, depending on the treatment dosage of EMS and the mutation discrimination platform used for genotype analysis. We implemented a three-dimensional DNA pooling strategy combined with multiplexed amplicon sequencing to create a highly efficient and cost-effective TILLING (targeting induced locus lesion in genomes) platform in barley. Mutations were successfully identified from 72 mixed amplicons within a DNA pool containing 64 individual mutants and from 56 mixed amplicons within a pool containing 144 individuals. We discovered abundant allelic mutants for dozens of genes, including the barley Green Revolution contributor gene Brassinosteroid insensitive 1 (BRI1). As a proof of concept, we rapidly determined the causal gene responsible for a chlorotic mutant by following the MutMap strategy, demonstrating the value of this resource to support forward and reverse genetic studies in barley.

PMID: 35605197