EMBO J , IF:9.889 , 2021 Jan , V40 (2) : Pe104559 doi: 10.15252/embj.2020104559
The calcium transporter ANNEXIN1 mediates cold-induced calcium signaling and freezing tolerance in plants.
State Key Laboratory of Plant Physiology and Biochemistry, College of Biological Sciences, China Agricultural University, Beijing, China.; Institute of Plant Stress Biology, Collaborative Innovation Center of Crop Stress Biology, Henan University, Kaifeng, China.; Department of Plant Sciences, University of Cambridge, Cambridge, UK.; Department of Biosciences, Durham University, Durham, UK.
The transient elevation of cytosolic free calcium concentration ([Ca(2+) ]cyt ) induced by cold stress is a well-established phenomenon; however, the underlying mechanism remains elusive. Here, we report that the Ca(2+) -permeable transporter ANNEXIN1 (AtANN1) mediates cold-triggered Ca(2+) influx and freezing tolerance in Arabidopsis thaliana. The loss of function of AtANN1 substantially impaired freezing tolerance, reducing the cold-induced [Ca(2+) ]cyt increase and upregulation of the cold-responsive CBF and COR genes. Further analysis showed that the OST1/SnRK2.6 kinase interacted with and phosphorylated AtANN1, which consequently enhanced its Ca(2+) transport activity, thereby potentiating Ca(2+) signaling. Consistent with these results and freezing sensitivity of ost1 mutants, the cold-induced [Ca(2+) ]cyt elevation in the ost1-3 mutant was reduced. Genetic analysis indicated that AtANN1 acts downstream of OST1 in responses to cold stress. Our data thus uncover a cascade linking OST1-AtANN1 to cold-induced Ca(2+) signal generation, which activates the cold response and consequently enhances freezing tolerance in Arabidopsis.
PMID: 33372703
Plant Cell Environ , IF:6.362 , 2021 Jan doi: 10.1111/pce.13993
Maize metabolome and proteome responses to controlled cold stress partly mimic early-sowing effects in the field and differ from those of Arabidopsis.
Biologie du Fruit et Pathologie, UMR 1332, Centre INRAE de Nouvelle Aquitaine-Bordeaux, INRAE, Univ., Villenave d'Ornon, France.; INRAE, CNRS, AgroParisTech, GQE-Le Moulon, Univ. Paris-Saclay, Gif-sur-Yvette, France.; PAPPSO, doi:10.15454/1.5572393176364355E12, GQE-Le Moulon, Gif-sur-Yvette, France.; PMB-Metabolome, INRAE, 2018, Bordeaux Metabolome, doi:10.15454/1.5572412770331912E12, MetaboHUB, PHENOME, IBVM, Centre INRAE de Nouvelle Aquitaine-Bordeaux, Villenave d'Ornon, France.; INRAE, UE GCIE, Estrees-Mons, Peronne, France.; INRAE, Univ. Liege, Univ. Lille, Univ. Picardie Jules Verne, BioEcoAgro, Peronne, France.
In Northern Europe, sowing maize one-month earlier than current agricultural practices may lead to moderate chilling damage. However, studies of the metabolic responses to low, non-freezing, temperatures remain scarce. Here, genetically-diverse maize hybrids (Zea mays, dent inbred lines crossed with a flint inbred line) were cultivated in a growth chamber at optimal temperature and then three decreasing temperatures for 2 days each, as well as in the field. Leaf metabolomic and proteomic profiles were determined. In the growth chamber, 50% of metabolites and 18% of proteins changed between 20 and 16 degrees C. These maize responses, partly differing from those of Arabidopsis to short-term chilling, were mapped on genome-wide metabolic maps. Several metabolites and proteins showed similar variation for all temperature decreases: seven MS-based metabolite signatures and two proteins involved in photosynthesis decreased continuously. Several increasing metabolites or proteins in the growth-chamber chilling conditions showed similar trends in the early-sowing field experiment, including trans-aconitate, three hydroxycinnamate derivatives, a benzoxazinoid, a sucrose synthase, lethal leaf-spot 1 protein, an allene oxide synthase, several glutathione transferases and peroxidases. Hybrid groups based on field biomass were used to search for the metabolite or protein responses differentiating them in growth-chamber conditions, which could be of interest for breeding.
PMID: 33410508
Plant J , IF:6.141 , 2021 Jan doi: 10.1111/tpj.15170
Ethylene increases the cold tolerance of apple via the MdERF1B-MdCIbHLH1 regulatory module.
State Key Laboratory of Crop Biology, Shandong Agricultural University, Tai'an, Shandong, China.; College of Horticulture Sciences, Shandong Agricultural University, Tai'an, Shandong, China.
Cold stress has always been a major abiotic factor affecting the yield and quality of temperate fruit crops. Ethylene plays a critical regulatory role in the cold stress response, but the underlying molecular mechanisms remains elusive. Here, we revealed that ethylene positively modulates apple responses to cold stress. Treatments with 1-aminocyclopropane-1-carboxylate (an ethylene precursor) and aminoethoxyvinylglycine (an ethylene biosynthesis inhibitor) respectively increased and decreased the cold tolerance of apple seedlings. Consistent with the positive effects of ethylene on cold stress responses, a low-temperature treatment rapidly induced ethylene release and the expression of MdERF1B, which encodes an ethylene signaling activator, in apple seedlings. The overexpression of MdERF1B significantly increased the cold tolerance of apple plant materials (seedlings and calli) and Arabidopsis thaliana seedlings. A qRT-PCR analysis indicated that MdERF1B upregulates the expression of the cold-responsive gene MdCBF1 in apple seedlings. Moreover, MdCIbHLH1, which functions upstream of CBF-dependent pathways, enhanced the binding of MdERF1B to target gene promoters as well as the consequent transcriptional activation. The stability of MdERF1B-MdCIbHLH1 was affected by cold stress and ethylene. Furthermore, MdERF1B interacted with the promoters of two genes critical for ethylene biosynthesis, MdACO1 and MdERF3. The resulting upregulated expression of these genes promoted ethylene production. However, the downregulated MdCIbHLH1 expression in MdERF1B-overexpressing apple calli significantly inhibited ethylene production. These findings imply that MdERF1B-MdCIbHLH1 is a potential regulatory module that integrates the cold and ethylene signaling pathways in apple.
PMID: 33497017
Biotechnol Biofuels , IF:4.815 , 2021 Jan , V14 (1) : P22 doi: 10.1186/s13068-021-01881-6
AP2/ERF and R2R3-MYB family transcription factors: potential associations between temperature stress and lipid metabolism in Auxenochlorella protothecoides.
State Key Laboratory of Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing, 100193, China.; State Key Laboratory of Molecular Developmental Biology, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, 100101, China.; University of Chinese Academy of Sciences, Beijing, 100049, China.; State Key Laboratory of Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing, 100193, China. yangjsh1999@163.com.
BACKGROUND: Both APETALA2/Ethylene Responsive Factor (AP2/ERF) superfamily and R2R3-MYB family were from one of the largest diverse families of transcription factors (TFs) in plants, and played important roles in plant development and responses to various stresses. However, no systematic analysis of these TFs had been conducted in the green algae A. protothecoides heretofore. Temperature was a critical factor affecting growth and lipid metabolism of A. protothecoides. It also remained largely unknown whether these TFs would respond to temperature stress and be involved in controlling lipid metabolism process. RESULTS: Hereby, a total of six AP2 TFs, six ERF TFs and six R2R3-MYB TFs were identified and their expression profiles were also analyzed under low-temperature (LT) and high-temperature (HT) stresses. Meanwhile, differential adjustments of lipid pathways were triggered, with enhanced triacylglycerol accumulation. A co-expression network was built between these 18 TFs and 32 lipid-metabolism-related genes, suggesting intrinsic associations between TFs and the regulatory mechanism of lipid metabolism. CONCLUSIONS: This study represented an important first step towards identifying functions and roles of AP2 superfamily and R2R3-MYB family in lipid adjustments and response to temperature stress. These findings would facilitate the biotechnological development in microalgae-based biofuel production and the better understanding of photosynthetic organisms' adaptive mechanism to temperature stress.
PMID: 33451355
BMC Genomics , IF:3.594 , 2021 Jan , V22 (1) : P51 doi: 10.1186/s12864-020-07365-5
Proteome-wide and lysine crotonylation profiling reveals the importance of crotonylation in chrysanthemum (Dendranthema grandiforum) under low-temperature.
Department of Ornamental Horticulture, Sichuan Agricultural University, 211 Huimin Road, Wenjiang District, Chengdu, Sichuan, 611130, People's Republic of China.; Department of Ornamental Horticulture, Sichuan Agricultural University, 211 Huimin Road, Wenjiang District, Chengdu, Sichuan, 611130, People's Republic of China. qinglinliu@126.com.
BACKGROUND: Low-temperature severely affects the growth and development of chrysanthemum which is one kind of ornamental plant well-known and widely used in the world. Lysine crotonylation is a recently identified post-translational modification (PTM) with multiple cellular functions. However, lysine crotonylation under low-temperature stress has not been studied. RESULTS: Proteome-wide and lysine crotonylation of chrysanthemum at low-temperature was analyzed using TMT (Tandem Mass Tag) labeling, sensitive immuno-precipitation, and high-resolution LC-MS/MS. The results showed that 2017 crotonylation sites were identified in 1199 proteins. Treatment at 4 degrees C for 24 h and - 4 degrees C for 4 h resulted in 393 upregulated proteins and 500 downregulated proteins (1.2-fold threshold and P < 0.05). Analysis of biological information showed that lysine crotonylation was involved in photosynthesis, ribosomes, and antioxidant systems. The crotonylated proteins and motifs in chrysanthemum were compared with other plants to obtain orthologous proteins and conserved motifs. To further understand how lysine crotonylation at K136 affected APX (ascorbate peroxidase), we performed a site-directed mutation at K136 in APX. Site-directed crotonylation showed that lysine decrotonylation at K136 reduced APX activity, and lysine complete crotonylation at K136 increased APX activity. CONCLUSION: In summary, our study comparatively analyzed proteome-wide and crotonylation in chrysanthemum under low-temperature stress and provided insights into the mechanisms of crotonylation in positively regulated APX activity to reduce the oxidative damage caused by low-temperature stress. These data provided an important basis for studying crotonylation to regulate antioxidant enzyme activity in response to low-temperature stress and a new research ideas for chilling-tolerance and freezing-tolerance chrysanthemum molecular breeding.
PMID: 33446097
J Biotechnol , IF:3.503 , 2021 Jan , V327 : P97-105 doi: 10.1016/j.jbiotec.2021.01.003
Expression of ice recrystallization inhibition protein in transgenic potato lines associated with reduced electrolyte leakage and efficient recovery post freezing injury.
Centre of Excellence in Molecular Biology, University of the Punjab, Lahore, Pakistan.; Department of Microbiology, BUITEMS, Quetta, Pakistan.; Institute of Agricultural Sciences (IAGS), University of the Punjab Lahore-Pakistan, Pakistan.; Centre of Excellence in Molecular Biology, University of the Punjab, Lahore, Pakistan; School of Biological Sciences, University of the Punjab, Pakistan. Electronic address: bushratabassum.sbs@pu.edu.pk.
Potato (Solanum tuberosum L.) is considered to be frost-susceptible as short spells of frost can reduce the tuber yield and quality. Ice recrystallization inhibition (IRI) protein helps prevent growth of ice crystals in the cell apoplast during frost and help prevent damage associated with freezing stress. In this study, we investigated the in planta potential of Lolium perenne derived IRI3 transgene in improving the tolerance of transgenic potato lines for freezing stress. The codon optimized IRI3 transgene was introduced into potato cultivar Diamant through Agrobacterium mediated transformation. Three transgenic potato lines were successfully generated which were confirmed for transgene insertion and genomic integration by polymerase chain reaction and Southern blot. It was evident that the IRI3 transcript decreased in initial 24 h of cold stress treatment while the IRI3 mRNA expression up regulated in subsequent hours of cold treatment with maximum increase to 20 folds at 96 h post stress. A similar trend was also revealed in ion-leakage assay which showed that during cold stress, the transgenic potato lines depicted reduced ion leakage of 14-22% as compared to non-transgenic control plants. Further, the generated transgenic potato lines were tolerant to the frost spell in quarantine field conditions as compared to the non-transgenic potato lines. Additionally, the transgenic lines exhibited efficient recovery post frost injury in field conditions. The biochemical profiles of chlorophyll, proline and higher levels of antioxidant enzyme (superoxide dismutase, Catalase) activity and malondialdehyde levels showed that despite the phenotypic impact of low temperature, the transgenic potato lines quickly adjusted to maintain their cellular homeostasis post freezing stress by increasing the antioxidant defenses. This study suggests that up regulation of IRI3 transcript and regulatory network of cold stress response in transgenic potato lines improve frost tolerance and help stabilize yield in cultivated potato.
PMID: 33450348
BMC Plant Biol , IF:3.497 , 2021 Jan , V21 (1) : P21 doi: 10.1186/s12870-020-02811-8
Genome-wide identification and analysis of DNA methyltransferase and demethylase gene families in Dendrobium officinale reveal their potential functions in polysaccharide accumulation.
Key Laboratory of South China Agricultural Plant Molecular Analysis and Genetic Improvement, South China Botanical Garden, Chinese Academy of Sciences, Guangzhou, 510650, China.; Economic Botany, Core Botanical Gardens, Chinese Academy of Sciences, Guangzhou, 510650, China.; Independent researcher, P. O. Box 7, Miki-cho post office, Ikenobe 3011-2, Miki-cho, Kagawa-ken, 761-0799, Japan.; Biodata Biotechnology Co. Ltd, Hefei, 230031, China.; Key Laboratory of South China Agricultural Plant Molecular Analysis and Genetic Improvement, South China Botanical Garden, Chinese Academy of Sciences, Guangzhou, 510650, China. duanj@scib.ac.cn.; Economic Botany, Core Botanical Gardens, Chinese Academy of Sciences, Guangzhou, 510650, China. duanj@scib.ac.cn.
BACKGROUND: DNA methylation is a conserved and important epigenetic modification involved in the regulation of numerous biological processes, including plant development, secondary metabolism, and response to stresses. However, no information is available regarding the identification of cytosine-5 DNA methyltransferase (C5-MTase) and DNA demethylase (dMTase) genes in the orchid Dendrobium officinale. RESULTS: In this study, we performed a genome-wide analysis of DoC5-MTase and DodMTase gene families in D. officinale. Integrated analysis of conserved motifs, gene structures and phylogenetic analysis showed that eight DoC5-MTases were divided into four subfamilies (DoCMT, DoDNMT, DoDRM, DoMET) while three DodMTases were divided into two subfamilies (DoDML3, DoROS1). Multiple cis-acting elements, especially stress-responsive and hormone-responsive ones, were found in the promoter region of DoC5-MTase and DodMTase genes. Furthermore, we investigated the expression profiles of DoC5-MTase and DodMTase in 10 different tissues, as well as their transcript abundance under abiotic stresses (cold and drought) and at the seedling stage, in protocorm-like bodies, shoots, and plantlets. Interestingly, most DoC5-MTases were downregulated whereas DodMTases were upregulated by cold stress. At the seedling stage, DoC5-MTase expression decreased as growth proceeded, but DodMTase expression increased. CONCLUSIONS: These results provide a basis for elucidating the role of DoC5-MTase and DodMTase in secondary metabolite production and responses to abiotic stresses in D. officinale.
PMID: 33407149
BMC Plant Biol , IF:3.497 , 2021 Jan , V21 (1) : P10 doi: 10.1186/s12870-020-02746-0
Cloning and characterization of KoOsmotin from mangrove plant Kandelia obovata under cold stress.
State Key Laboratory of Tropical Oceanography, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou, 510301, China.; Southern Marine Science and Engineering Guangdong Laboratory, Guangzhou, 511458, China.; Innovation Academy of South China Sea Ecology and Environmental Engineering, Chinese Academy of Sciences, Guangzhou, 510301, China.; State Key Laboratory of Tropical Oceanography, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou, 510301, China. yswang@scsio.ac.cn.; Southern Marine Science and Engineering Guangdong Laboratory, Guangzhou, 511458, China. yswang@scsio.ac.cn.; Innovation Academy of South China Sea Ecology and Environmental Engineering, Chinese Academy of Sciences, Guangzhou, 510301, China. yswang@scsio.ac.cn.; State Key Laboratory of Tropical Oceanography, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou, 510301, China. chenghao@scsio.ac.cn.; Southern Marine Science and Engineering Guangdong Laboratory, Guangzhou, 511458, China. chenghao@scsio.ac.cn.; Innovation Academy of South China Sea Ecology and Environmental Engineering, Chinese Academy of Sciences, Guangzhou, 510301, China. chenghao@scsio.ac.cn.; College of Life Science and Technology, Jinan University, Guangzhou, 510632, China.; Root Biology Center, Fujian Agriculture and Forestry University, Fuzhou, 350002, China.
BACKGROUND: Low temperature is a major abiotic stress that seriously limits mangrove productivity and distribution. Kandelia obovata is the most cold-resistance specie in mangrove plants, but little is known about the molecular mechanism underlying its resistance to cold. Osmotin is a key protein associated with abiotic and biotic stress response in plants but no information about this gene in K. obovata was reported. RESULTS: In this study, a cDNA sequence encoding osmotin, KoOsmotin (GenBank accession no. KP267758), was cloned from mangrove plant K. obovata. The KoOsmotin protein was composed of 221 amino acids and showed a calculated molecular mass of 24.11 kDa with pI 4.92. The KoOsmotin contained sixteen cysteine residues and an N-terminal signal peptide, which were common signatures to most osmotins and pathogenesis-related 5 proteins. The three-dimensional (3D) model of KoOsmotin, contained one alpha-helix and eleven beta-strands, was formed by three characteristic domains. Database comparisons of the KoOsmotin showed the closest identity (55.75%) with the osmotin 34 from Theobroma cacao. The phylogenetic tree also revealed that the KoOsmotin was clustered in the branch of osmotin/OLP (osmotin-like protien). The KoOsmotin protein was proved to be localized to both the plasma membrane and cytoplasm by the subcellular localization analysis. Gene expression showed that the KoOsmotin was induced primarily and highly in the leaves of K. obovata, but less abundantly in stems and roots. The overexpressing of KoOsmotin conferred cold tolerance in Escherichia coli cells. CONCLUSION: As we known, this is the first study to explore the osmotin of K. obovata. Our study provided valuable clues for further exploring the function of KoOsmotin response to stress.
PMID: 33407136
Planta , IF:3.39 , 2021 Jan , V253 (1) : P9 doi: 10.1007/s00425-020-03521-z
Genetic analysis of freezing tolerance in camelina [Camelina sativa (L.) Crantz] by diallel cross of winter and spring biotypes.
Department of Plant Breeding and Biotechnology, Sari Agricultural Sciences and Natural Resources University (SANRU), P.O. Box 576, Sari, Iran. j.soorni@sanru.ac.ir.; Department of Systems Biology, Agricultural Biotechnology Research Institute of Iran (ABRII), Agricultural Research, Education and Extension Organization (AREEO), P.O. Box 31535-1897, Karaj, Iran. j.soorni@sanru.ac.ir.; Department of Plant Breeding and Biotechnology, Sari Agricultural Sciences and Natural Resources University (SANRU), P.O. Box 576, Sari, Iran.; Department of Agronomy and Plant Breeding, Razi University, P.O. Box 85438-67156, Kermanshah, Iran.; Genetics and Agricultural Biotechnology Institute of Tabarestan (GABIT), Sari Agricultural Sciences and Natural Resources University (SANRU), P.O. Box 576, Sari, Iran.
MAIN CONCLUSION: Camelina biotypes had different responses to freezing stress, which was mainly inherited by additive gene effects and can be reliably used in breeding programs and for a better understanding of freezing tolerance mechanisms in camelina plants. Camelina [Camelina sativa (L.) Crantz] is a frost-tolerant oilseed plant that is cultivated as an autumn crop in semi-arid regions. However, camelina establishment in these areas is limited by low temperatures in winter that results in decreased seed yield. In the present study, genetic basis of freezing tolerance (FT) in spring and winter biotypes of camelina was analyzed at seedling stage using a diallel cross experiment. The parents consisted of two winter doubled haploid (DH) lines with high (DH34 and DH31), two spring lines with medium (DH19 and DH26), and two spring lines with low FT (DH08 and DH91). For this purpose, the parents along with F1 entries were subjected to freezing stress and survival percentage, electrolyte leakage, and lethal temperature for 50% mortality (LT50) of the lines were measured. Results showed that although both additive and non-additive effects of the genes determine the FT, further analyses indicated that it was mainly controlled by the additive effects. Therefore, selection-based methods may be more efficient for improving FT in camelina genotypes. The results of specific combining ability (SCA) and heterosis analysis among various DH lines suggested that more tolerant cultivars of camelina could be developed by targeted crossings. When a tolerant winter line and a susceptible spring line were crossed, their progenies showed a higher FT compared with the progenies of a cross between two susceptible spring lines indicating FT is controlled by additive effects of the genes in camelina plants. These findings provided new insight into the genetic basis of freezing-related traits in camelina and could be used for more sophisticated breeding programs.
PMID: 33389162
Gene , IF:2.984 , 2021 Jan , V764 : P145097 doi: 10.1016/j.gene.2020.145097
Cold-regulated gene LeCOR413PM2 confers cold stress tolerance in tomato plants.
College of Life Science, Shihezi University, Shihezi 832000, China.; College of Life Science, Shihezi University, Shihezi 832000, China. Electronic address: jianboz9@sina.com.
Tomato (Lycopersicon esculentum Mill) is an important food plant that has been used as a model plant in genetic evolution and molecular biology research. The plant is originated from the tropics; thus, it is sensitive to cold. Its growth and development can be easily affected by cold stress. In this study, cold-regulated gene LeCOR413PM2 was cloned from tomato leaves and then used to generate two types of transgenic tomato plants: LeCOR413PM2-overexpressing transgenic plants and RNA-interference-expressing transgenic plants. The functions and expression of LeCOR413PM2 gene in response to cold stress were subsequently assessed. The results showed that LeCOR413PM2 localized in the plasma membrane. Expression of LeCOR413PM2 gene in the leaf of transgenic tomato plant was highest compared to that in other organs (i.e., root, stem, flower and fruit); it was elevated when plants were treated with cold stress. Overexpression of LeCOR413PM2 gene was found to not only reduce damage to cell membrane, accumulation of ROS, and photoinhibition of PSII, but also maintain high activity of antioxidant enzymes and content of osmotic regulators. The results also reveal that high activities of antioxidant enzymes were caused by the up-regulation of their gene expressions. This study demonstrates that the overexpression of LeCOR413PM2 could increase cold tolerance of transgenic tomato plants, while the suppressed expression of LeCOR413PM2 by RNA interference could increase the sensitivity of plants to cold.
PMID: 32866589
Plants (Basel) , IF:2.762 , 2021 Jan , V10 (1) doi: 10.3390/plants10010104
CRISPR/Cas9-Mediated Knockout of HOS1 Reveals Its Role in the Regulation of Secondary Metabolism in Arabidopsis thaliana.
Federal Scientific Center of the East Asia Terrestrial Biodiversity, Far Eastern Branch, the Russian Academy of Sciences, 690022 Vladivostok, Russia.
In Arabidopsis, the RING finger-containing E3 ubiquitin ligase HIGH EXPRESSION OF OSMOTICALLY RESPONSIVE GENES 1 (HOS1) functions as a main regulator of the cold signaling. In this study, CRISPR/Cas9-mediated targeted mutagenesis of the HOS1 gene in the first exon was performed. DNA sequencing showed that frameshift indels introduced by genome editing of HOS1 resulted in the appearance of premature stop codons, disrupting the open reading frame. Obtained hos1(Cas9) mutant plants were compared with the SALK T-DNA insertion mutant, line hos1-3, in terms of their tolerance to abiotic stresses, accumulation of secondary metabolites and expression levels of genes participating in these processes. Upon exposure to cold stress, enhanced tolerance and expression of cold-responsive genes were observed in both hos1-3 and hos1(Cas9) plants. The hos1 mutation caused changes in the synthesis of phytoalexins in transformed cells. The content of glucosinolates (GSLs) was down-regulated by 1.5-times, while flavonol glycosides were up-regulated by 1.2 to 4.2 times in transgenic plants. The transcript abundance of the corresponding MYB and bHLH transcription factors, which are responsible for the regulation of secondary metabolism in Arabidopsis, were also altered. Our data suggest a relationship between HOS1-regulated downstream signaling and phytoalexin biosynthesis.
PMID: 33419060
PLoS One , IF:2.74 , 2021 , V16 (1) : Pe0243292 doi: 10.1371/journal.pone.0243292
Comparative proteomics analysis reveals the molecular mechanism of enhanced cold tolerance through ROS scavenging in winter rapeseed (Brassica napus L.).
Gansu Provincial Key Laboratory of Aridland Crop Science, College of Agronomy, Gansu Agricultural University, Lanzhou, China.
Two winter rapeseed cultivars, "NS" (cold tolerant) and "NF" (cold sensitive), were used to reveal the morphological, physiological, and proteomic characteristics in leaves of plants after treatment at -4 degrees C for 12 h(T1) and 24 h(T2), and at room temperature(T0), to understand the molecular mechanisms of cold tolerance. Antioxidant activity and osmotic adjustment ability were higher, and plasma membrane injury was less obvious, in NS than in NF under cold stress. We detected different abundant proteins (DAPs) related to cold tolerance in winter rapeseed through data-independent acquisition (DIA). Compared with NF, A total of 1,235 and 1,543 DAPs were identified in the NSs under T1 and T2, respectively. Compared with NF, 911 proteins were more abundant in NS only after cold treatment. Some of these proteins were related to ROS scavenging through four metabolic pathways: lysine degradation; phenylalanine, tyrosine, and tryptophan; flavonoid biosynthesis; and ubiquinone and other terpenoid-quinone biosynthesis. Analysis of these proteins in the four candidate pathways revealed that they were rapidly accumulated to quickly enhance ROS scavenging and improve the cold tolerance of NS. These proteins were noticeably more abundant during the early stage of cold stress, which was critical for avoiding ROS damage.
PMID: 33434207
Oecologia , IF:2.654 , 2021 Jan doi: 10.1007/s00442-020-04839-x
Carbon allocation to growth and storage depends on elevation provenance in an herbaceous alpine plant of Mediterranean climate.
ECOBIOSIS, Departamento de Botanica, Facultad de Ciencias Naturales y Oceanograficas, Universidad de Concepcion, Casilla 160-C, Concepcion, Chile. clau.m.reyes@gmail.com.; Instituto de Ecologia y Biodiversidad (IEB), Casilla 653, Santiago, Chile. clau.m.reyes@gmail.com.; Centro de Investigacion en Ecosistemas de la Patagonia (CIEP), Moraleda 16, Coyhaique, Chile.; ECOBIOSIS, Departamento de Botanica, Facultad de Ciencias Naturales y Oceanograficas, Universidad de Concepcion, Casilla 160-C, Concepcion, Chile.; Instituto de Ecologia y Biodiversidad (IEB), Casilla 653, Santiago, Chile.
It is unclear whether the frequently observed increase in non-structural carbohydrates (NSC) in plants exposed to low temperatures or drought reflects a higher sensitivity of growth than photosynthesis in such conditions (i.e. sink limitation), or a prioritization of carbon (C) allocation to storage. Alpine areas in Mediterranean-type climate regions are characterized by precipitation increases and temperature decreases with elevation. Thus, alpine plants with wide elevational ranges in Mediterranean regions may be good models to examine these alternative hypotheses. We evaluated storage and growth during experimental darkness and re-illumination in individuals of the alpine plant Phacelia secunda from three elevations in the Andes of central Chile. We hypothesized that storage is prioritized regarding growth in plants of both low- and high elevations where drought and cold stress are greatest, respectively. We expected that decreases in NSC concentrations during darkness should be minimal and, more importantly, increases in NSC after re-illumination should be higher than increases in biomass. We found that darkness caused a significant decrease in NSC concentrations of both low- and high-elevation plants, but the magnitude of the decrease was lower in the latter. Re-illumination caused higher increase in NSC concentration than in biomass in both low- and high-elevation plants (1.5- and 1.9-fold, respectively). Our study shows that C allocation in Phacelia secunda reflects ecotypic differences among elevation provenances and suggests that low temperature, but not drought, favours C allocation to storage over growth after severe C limitation.
PMID: 33459865
Funct Plant Biol , IF:2.617 , 2021 Jan doi: 10.1071/FP20046
Identification and characterisation of cold stress-related proteins in Oryza rufipogon at the seedling stage using label-free quantitative proteomic analysis.
In this study, label-free quantitative proteomics were used to study cold stress-related proteins in Dongxiang wild rice (Oryza rufipogon Griff., DWR) and cold sensitive cultivated rice 'Xieqingzao B'(Oryza sativa L. ssp. indica cv., XB). The results demonstrated the presence of 101 and 216 differentially expressed proteins (DEPs) were detected in DWR and XB, respectively, after cold stress. Bioinformatics analysis showed that DWR and XB differed significantly in their ability to scavenge reactive oxygen species (ROS) and regulate energy metabolism. Of the 101 DEPs of DWR, 46 DEPs related to differential expressed genes were also detected by transcriptome analysis. And 13 out of 101 DEPs were located in previous cold related quantitative trait loci (QTL). Quantitative real-time PCR analysis indicated that protein expression and transcription patterns were not similar in XB and DWR. Protein-protein interaction (PPI) network was constituted using the DEPs of DWR and XB, and the following three centre proteins were identified: Q8H3I3, Q9LDN2, and Q2QXR8. Next, we selected a centre protein and two of the 37 DEPs with high levels of differential expression (fold change >/= 2) were used for cloning and prokaryotic expression. We found that Q5Z9Q8 could significantly improve the cold tolerance of Escherichia coli.
PMID: 33487217
Cryobiology , IF:2.283 , 2021 Jan doi: 10.1016/j.cryobiol.2021.01.012
Differential proteome between ejaculate and epididymal sperm represents a key factor for sperm freezability in wild small ruminants.
Department of Animal Reproduction, Spanish National Institute for Agricultural and Food Research and Technology (INIA), Avda Puerta de Hierro km 5.9, 28040 Madrid, Spain; Department of Animal Breeding and Husbandry, Institute of Animal Science, Endenicher Allee 15, University of Bonn, 53115 Bonn, Germany; Department of Physiology, Faculty of Veterinary Science, International Excellence Campus for Higher Education and Research 'Campus Mare Nostrum', University of Murcia, Campus de Espinardo, 30100 Murcia, Spain.; Institute of Biochemistry and Molecular Biology, Medical Faculty, University of Bonn, Nussallee 11, 53115 Bonn, Germany.; Institute for Medical Biometry, Informatics and Epidemiology, Medical Faculty, University of Bonn, Venusberg-Campus 1, 53127 Bonn, Germany; Institute for Genomic Statistics and Bioinformatics, Medical Faculty, University of Bonn, Venusberg-Campus 1, 53127 Bonn, Germany.; Centre for Biotechnology and Plant Genomic, Polytechnic University of Madrid-National Institute for Agricultural and Food Research and Technology (UPM-INIA), Autopista M-40 Km 38, 28223 Madrid, Spain.; Department of Physiology, Faculty of Veterinary Science, International Excellence Campus for Higher Education and Research 'Campus Mare Nostrum', University of Murcia, Campus de Espinardo, 30100 Murcia, Spain.; Department of Animal Breeding and Husbandry, Institute of Animal Science, Endenicher Allee 15, University of Bonn, 53115 Bonn, Germany.; Department of Biomedical Sciences, Animal Reproduction and Biotechnology Laboratory, Colorado State University, 3105 Rampart Rd, 80521 Fort Collins, CO, USA.; Department of Animal Reproduction, Spanish National Institute for Agricultural and Food Research and Technology (INIA), Avda Puerta de Hierro km 5.9, 28040 Madrid, Spain. Electronic address: moreno@inia.es.
Epididymal sperm shows higher cryoresistance than ejaculated sperm. Although the sperm proteome seems to affect cell cryoresistance, studies aiming at identifying proteins involved in sperm freezing-tolerance are scarce. The aims of this study were to investigate differences of sperm freezability and proteome between epididymal and ejaculated sperm in three mountain ungulates: Iberian ibex, Mouflon and Chamois. Sperm samples were cryopreserved in straws by slow freezing. Tandem mass tag-labeled peptides from sperm samples were analyzed by high performance liquid chromatography coupled to a mass spectrometer in three technical replicates. The statistical analysis was done using the moderated t-test of the R package limma. Differences of freezability between both types of sperm were associated with differences of the proteome. Overall, epididymal sperm showed higher freezability than ejaculated sperm. Between 1490 and 1883 proteins were quantified in each species and type of sperm sample. Cross species comparisons revealed a total of 76 proteins that were more abundant in epididymal than in ejaculated sperm in the three species of study whereas 3 proteins were more abundant in ejaculated than epididymal sperm in the three species of study (adjusted P < 0.05; |log2| fold-change > 0.5). Many of the proteins that were associated with higher cryoresistance are involved in stress response and redox homeostasis. In conclusion, marked changes of sperm proteome were detected between epididymal and ejaculated sperm. This work contributes to update the sperm proteome of small ruminants and to identify candidate markers of sperm freezability.
PMID: 33485896
Plant Signal Behav , IF:1.671 , 2021 Jan , V16 (1) : P1845048 doi: 10.1080/15592324.2020.1845048
Molecular cloning and functional characterization of GmAAPTs from soybean (Glycine max).
College of Life Sciences, State Key Laboratory of Crop Genetics and Germplasm Enhancement, Nanjing Agricultural University , Nanjing, P.R.China.; Institute of Botany, Jiangsu Province and Chinese Academy of Sciences (Nanjing Botanical Garden Mem. Sun Yat-Sen) , Nanjing, P.R.China.; College of Agriculture, State Key Laboratory of Crop Genetics and Germplasm Enhancement, Nanjing Agricultural University , Nanjing, P.R.China.; Institute of Industrial Crops, Jiangsu Academy of Agricultural Sciences , Nanjing, P.R.China.
Aminoalcoholphosphotransferase (AAPT) utilizes diacylglycerols and cytidine diphosphate-choline/ethanolamine as substrates for the synthesis of phosphatidylcholine (PC)/phosphatidylethanolamine (PE). Plant AAPTs involved in phospholipid metabolism mediate diverse physiological processes; however, little is known about their functions in triacylglycerol (TAG) metabolism and seed germination. In the present study, we isolated and characterized two AAPTs, GmAAPT1 and GmAAPT2, from soybean (Glycine max). GmAAPT1 and GmAAPT2 exhibited strong similarity in their amino acid contents and expression patterns, and both were found to localize to the endoplasmic reticulum and Golgi apparatus. In vitro enzymatic analyses showed that GmAAPT1 and GmAAPT2 contributed to PC and PE synthesis and exhibited choline/ethanolamine phosphotransferase-like enzymatic properties. The overexpression of GmAAPT1 and GmAAPT2 in Arabidopsis led to reduced levels of seed TAG and polyunsaturated fatty acids and decreased seed germination under freezing stress. Together, these findings suggest that GmAAPTs mediate TAG metabolism and negatively regulate seed freezing tolerance.
PMID: 33164676