摘要
大豆(Gycine max L.)是我国重要的经济作物之一。干旱、盐渍是造成农作物减产、耕作面积减少的两大重要原因。土壤盐渍化对大豆的生产影响非常大,利用现代生物技术提高大豆的耐盐性,是解决相关问题的一种有效途径。
本研究利用合作单位提供的大豆耐盐主效QTL定位信息,通过遗传基因组学分析,将大豆耐盐主效QTL定位于三号染色体的39.69Mbp—44.06Mbp区间,利用Softberry预测出该区段包含约746个ORF；再由BLAST对所有ORF进行了功能分析,发现该区段包含19个可能与耐盐密切有关的基因。在大豆合丰39幼苗经150mM氯化钠溶液处理后,利用实时荧光定量PCR对这19个候选基因进行时空表达模式分析,发现盐胁迫后有4个候选基因变化明显,即4-442在根和叶中表达均显著上调,7-316、17-641、19-737等候选基因在根中表达显著上调,而在叶中表达显著下调。
下一步将对4-442、7-316、17-641、19-737等候选基因进行全长克隆,并利用农杆菌介导转化拟南芥、水稻,进一步研究这四个基因的功能。
此外,本课题组前期发现了一个对盐胁迫有应答的候选基因Gm179,对Gm179完成了全长克隆、测序、转基因载体构建等工作,为下阶段研究打下了扎实的基础。
Soybean (Gycine max L.) is one of important economic crops of China. Drought and salinization are the two major causes leads to cultivable land and yield decrease. Modern biotechnology is an effective way to improve the salt tolerance of soybean which is a promising approach to solve the problem of salinization.
According to the fine mapping information of soybean salt tolerance main QTL from co-operation, salt tolerant main QTL was mapped in the interval of 39.69Mbp-44.06Mbp on the third chromosome by genetical genomics analysis. About 746 ORFs were predicted from this fragment by FGENESH on Softberry. All ORFs'function was predicted through BLAST analysis. And about 19 ORFs were identified which may contribute to soybean salt tolerance trait. Hefeng 39 seedlings were treated with 150mM NaCl, and all 19 candidate genes'time-spatial expression patterns were analyzed by real time quantitative PCR. After salt stress, gene expression of ORF4-442 was up-regulated significantly both in roots and leaves; meanwhile, ORF7-316, ORF17-641 and ORF19-737 were only up-regulated in roots and down-regulated in leaf significantly.
Full length cDNA of 4-442,7-316,17-641,19-737 will be cloned, and transformed into Arabidopsis and rice by Agrobacterium-mediated transformation to further verify these four genes'function in vivo in following work.
In addition, Gm179, one former identified salt tolerance candidate gene in our lab, was cloned, sequenced and transgenic constructor was constructed, which has laid a solid foundation for further functional analysis of Gm179 in vivo.
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