拟南芥耐盐突变体的筛选和相关基因的研究
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摘要
土地盐碱化限制了植物的生长,降低了农作物的产量,严重危害了我国农业的可持续性发展。因此,人们投入了大量的精力来研究植物对外界盐碱胁迫的各种耐受机制。植物体内的耐盐机制纵横交错,非常复杂。
我们筛选得到了两个耐受200mMNaCl萌发的拟南芥突变体AT6和AT13。AT6突变体是基因ATlg73660的功能缺失突变体,它在种子萌发阶段和萌发后的幼苗生长阶段均具有耐盐表型。根据预测ATlg73660编码拟南芥Raf类的MAPKKK。在种子萌发阶段,ATlg73660基因的三个功能敲除突变体对于ABA、GA和乙烯的效应程度和野生型没有差异。总之,研究表明拟南芥中ATlg73660基因的编码产物对盐胁迫的耐受机制起到负调控作用。
AT13突变体中T-DNA的插入导致AT3g58110基因的敲除。通过萌发阶段ABA敏感度的实验分析,我们推测AT3g58110基因可能通过ABA介导的途径负调节种子的耐盐萌发。另外AT13突变体在正常生长状况下的晚花和叶片深绿的表型也说明AT3g58110基因的编码蛋白是植物生长发育的调控因子。
Salinity greatly limits plant growth and crop productivity and poses an increasingly serious threat to the sustainability of agriculture. As a consequence, unremitting effort has been conducted to determine the adaptive mechanisms that plants have evolved to survive this abiotic stress. Salt tolerance mechanisms in plants are very complex and involve multiple parallel and interacting pathways.
Here, we reported two mutants that can germinate at 200 mM NaCl and designed as AT6 and AT13, respectively. AT6, a KO mutant of AT1g73660 exhibits salt tolerance phenotype during germination and seedling growth. AT1g73660 encodes a protein with similarity to Raf-like MAPKKKs. No difference in germination was observed between the mutants and the wildtype during the sensitivity assay to ABA, GA or ACC. Taken together, our results showed that AT1g73660 negatively regulated salt tolerance.
Cloning of the chromosomal DNA flanking the T-DNA in AT13 mutant revealed that the insertion had caused a knockout mutation in the gene AT3g58110. To confirm the gene-to-trait relationship of AT13 mutant, germination frequency of two other mutant alleles was observed in the presence of NaCl and ABA. All these assays showed that gene AT3g58110 negatively regulated the salt regulation of seed germination via the ABA pathway in Arabidopsis. In addition, the late-flowering phenotype and dark green leaves implied that the gene AT3g58110 product was also a regulator of plant development.
我们筛选得到了两个耐受200mMNaCl萌发的拟南芥突变体AT6和AT13。AT6突变体是基因ATlg73660的功能缺失突变体,它在种子萌发阶段和萌发后的幼苗生长阶段均具有耐盐表型。根据预测ATlg73660编码拟南芥Raf类的MAPKKK。在种子萌发阶段,ATlg73660基因的三个功能敲除突变体对于ABA、GA和乙烯的效应程度和野生型没有差异。总之,研究表明拟南芥中ATlg73660基因的编码产物对盐胁迫的耐受机制起到负调控作用。
AT13突变体中T-DNA的插入导致AT3g58110基因的敲除。通过萌发阶段ABA敏感度的实验分析,我们推测AT3g58110基因可能通过ABA介导的途径负调节种子的耐盐萌发。另外AT13突变体在正常生长状况下的晚花和叶片深绿的表型也说明AT3g58110基因的编码蛋白是植物生长发育的调控因子。
Salinity greatly limits plant growth and crop productivity and poses an increasingly serious threat to the sustainability of agriculture. As a consequence, unremitting effort has been conducted to determine the adaptive mechanisms that plants have evolved to survive this abiotic stress. Salt tolerance mechanisms in plants are very complex and involve multiple parallel and interacting pathways.
Here, we reported two mutants that can germinate at 200 mM NaCl and designed as AT6 and AT13, respectively. AT6, a KO mutant of AT1g73660 exhibits salt tolerance phenotype during germination and seedling growth. AT1g73660 encodes a protein with similarity to Raf-like MAPKKKs. No difference in germination was observed between the mutants and the wildtype during the sensitivity assay to ABA, GA or ACC. Taken together, our results showed that AT1g73660 negatively regulated salt tolerance.
Cloning of the chromosomal DNA flanking the T-DNA in AT13 mutant revealed that the insertion had caused a knockout mutation in the gene AT3g58110. To confirm the gene-to-trait relationship of AT13 mutant, germination frequency of two other mutant alleles was observed in the presence of NaCl and ABA. All these assays showed that gene AT3g58110 negatively regulated the salt regulation of seed germination via the ABA pathway in Arabidopsis. In addition, the late-flowering phenotype and dark green leaves implied that the gene AT3g58110 product was also a regulator of plant development.
引文
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