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HsfAld, a Protein Identified via FOX Hunting Using thellungiella salsuginea cDNAs Improves Heat Tolerance by Regulating Heat-Stress-Responsive Gene Expression

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Molecular Plant 2013年第2期6卷 411-422页

作者： Yukari Higashi Naohiko Ohama Tomoko Ishikawa Taku Katori Ayaka Shimura Kazuya Kusakabe Kazuko Yamaguchi-Shinozaki Junko Ishida Maho Tanaka Motoaki Seki Kazuo Shinozaki Yoichi Sakata Takahisa Hayashi Teruaki Taji Department of Bio-Science Tokyo University of Agriculture 1-1-1 Sakuragaoka Setagaya Tokyo 156-8502 Japan Graduate School of Agriculture and Life Sciences University of Tokyo Tokyo 113-8657 Japan RIKEN Yokohama Institute 1-7-22 Suehiro-cho Tsururni-ku Yokohama 230-0045 Japan

thellungiella salsuginea （formerly T. halophila）, a species closely related to Arabidopsis （Arabidopsis thali-ana）, is tolerant not only to high salt levels, but also to chilling, freezing, and ozone. Here, we report that T. salsuginea also shows greater heat tolerance than Arabidopsis. We identified T. salsuginea HsfAld （TsHsfAld） as a gene that can confer marked heat tolerance on Arabidopsis. TsHsfAld was identified via Full-length cDNA Over-eXpressing gene （FOX） hunt-ing from among a collection of heat-stress-related T. salsuginea cDNAs. Transgenic Arabidopsis overexpressing TsHsfAld showed constitutive up-regulation of many genes in the Arabidopsis AtHsfA1 regulon under normal growth tempera-ture. in Arabidopsis mesophyll protoplasts, TsHsfAld was localized in both the nucleus and the cytoplasm. TsHsfAld also interacted with AtHSP90, which negatively regulates AtHsfAls by forming HsfA1-HSP90 complexes in the cytoplasm. It is likely that the partial nuclear localization of TsHsfAld induced the expression of the AtHsfAld regulon in the transgenic plants at normal temperature. We also discovered that transgenic Arabidopsis plants overexpressing AtHsfAldwere more heat-tolerant than wild-type plants and up-regulated the expression of the HsfAld regulon, as was observed in TsHsfAld-overexpressing plants. We propose that the products of both TsHsfAld and AtHsfAld function as positive regulators of Arabidopsis heat-stress response and would be useful for the improvement of heat-stress tolerance in other plants.

关键词： thellungiella salsuginea heat-stress tolerance HsfA1 FOX hunting.

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Salt-responsive miRNA in root of thellungiella salsuginea

Salt-responsive miRNA in root of Thellungiella salsuginea

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从植物科学到农业发展——2012全国植物生物学大会

作者： Fei Gao,Jing Chen,Xin Sui,Yijun Zhou~* College of Life and Environmental Sciences,Minzu University of China,Beijing 100081,China.

MicroRNA(miRNA) has been administrated as a key regulatory component in plant stress adaptation.thellungiella salsuginea(halophila),a salt tolerant relative of Arabidopsis,is a good model used in plant salt tolerance studies,however,there is no report on identification and stress response profiling of miRNA in T.salsuginea.In the present study,Illumina sequencing generated two small RNA libraries using RNA isolated from high-salinity(SS) treated and untreated(CK) roots of T.salsuginea.Informatics analysis identified 49 conserved miRNA generated from 74 conserved miRNA precursors and 60 novel miRNA generated from 79 precursors,respectively.Comparing the reads of miRNA in CK and SS libraries identified 18 salt-responsive(change ratio > 1.5) conserved miRNA including 13 up-regulated(including miR160b,miR166-l,mirl71c-l,miR2111a,mir319-2,mir394,and mir396) and 5 down-regulated(including miR167-l miR395a,miR398a,and miR399b) conserved thellungiella miRNAs.Twenty-nine salt-responsive including 6 up-regulated and 23 down-regulated novel thellungiella miRNAs were also identified.Quantitative RT-PCRs were performed to validated these miRNA and expression profile.Target gene prediction and degradome sequencing revealed that these salt-responsive miRNA may participate in salt stress response in thellungiella root by regulating the gene expression of transcription factors like ARF,GRF,TCP,SCL,NF-YC,cell cycle related protein like CYCLIN A and enzymes like CSD and ATP sulphurylase.Finally,a network model regulating the miRNA mediated salt stress response in thellungiella root was proposed. Identification of miRNAs participating in stress response of T.salsuginea root will improve our understanding on molecular mechanisms of plant salt tolerance.

关键词： thellungiella salsuginea root salt stress miRNA

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Overexpression of a galactinol synthase,TsGOLS2,in Arabidopsis thaliana enhances tolerance to high salinity and osmotic stresses

Overexpression of a galactinol synthase,TsGOLS2,in Arabidops...

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从植物科学到农业发展——2012全国植物生物学大会

作者： Zhibin Sun~1,Zenglan Wang~1,Xingyun Qi~1,Pinghua Li,Chunxia Wu,Hui Zhang and Yanxiu Zhao~* Key Lab of Plant Stress Research,Life Science College of Shandong Normal University,88 Wenhua East Road,Jinan 250014,China.

Galactinol synthase(GOLS),the key enzyme in the synthesis of raffinose family oligosaccharides(RFOs),catalyzes the condensation of UDP-galactose with /nyo-inositol to produce galactinol as the sole donor for the synthesis of RFOs.RFOs have been implicated in mitigating effects of environmental stresses on plants.We have cloned a galactinol synthase gene from thellungiella salsuginea.The TsGOLS2 gene(1,322 bp) contained an open reading frame of 1011 bp and encodes 337 amino acids,with high homology to AtGolS2.TsGOLS2 was up-regulated by several abiotic stresses.Using molecular biological methods,we over-expressed TsGOLS2 in A.thaliana.The contents of galactinol and raffinose were significantly increased in transgenic plants.Salt-stressed transgenic Arabidopsis plants exhibited higher germination rate,photosynthesis ability and seedling growth.Following osmotic stress by high concentration of sorbitol,transgenic plants retained high germination rates and grew well during early development.These results indicated that over-expression of TsGOLS in A.thaliana improved the tolerance to high salinity and osmotic stress of transgenic plants.

关键词： thellungiella salsuginea TsGOLS2 overexpression galactinol raffinose stress tolerance.

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A mobile C2-domain protein confers salt tolerance in plant

A mobile C2-domain protein confers salt tolerance in plant

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从植物科学到农业发展——2012全国植物生物学大会

作者： Gang Li,Ran Xia,Yiyue Zhang,Huawei Zhang,Qi Xie~* Institute of Genetics and Developmental Biology,Chinese Academy of Sciences,Beijing 100101,China

A mobile C2-domain protein TsST03 has been isolated from thellungiella salsuginea through functional expression of random cDNA libraries of T.salsuginea in Arabidopsis due to its ability to increase the salt tolerance.The expression levels of TsST03 was found to be induced quickly by NaCl,drought,cold and heat stress as well as stress-related hormones including ABA,JA,ACC and SA,which suggests TsTh03 perhaps participates in the early response to these stresses.Studies of subcellular localization of TsTh03 in TsTh03-GFP transgenic Arabidopsis droved by the native promoter of TsTh03 revealed that TsTh03-GFP green fluorescence was uniformly dispersed throughout the cytosol,nucleus and plasma membrane.However,subject to salt stress,signals emitted from TsTh03-GFP disappeared from the nucleus and cytosol,and could only be detected on the plasma membrane.These results indicated the subcellular localization of TsTh03 was regulated by salt stress.Heterograft experiments in Arabidopsis revealed the protein of TsTh03 could participate in long distance translocation,by which the wild type part in chimera plant could be endowed with salt tolerant abilities.Peptide analysis of the TsTh03 sequence revealed that TsTh03 had the highest sequence similarity with C2 domain-containing proteins.Details will be presented and discussed.

关键词： thellungiella salsuginea salt tolerance C2-domain movement protein

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