摘要根瘤菌不但可以通过共生固氮提升大豆品质,还可提高大豆的耐盐能力,但其机制仍不明确。本研究中,我们采用qRT-PCR技术,研究NaCl胁迫下重要代谢通路上的关键调控基因的转录表达情况。实验结果表明:盐胁迫下的Union85140盐敏感大豆叶中转录因子MYB、CHS、CHI、CPM编码基因的转录水平都因根瘤菌施用呈现显著的上调趋势,尤其是CHI-1编码基因的上调趋势最为显著,达到了64705。8%,其次是CHS-3编码基因,达到了6065。8%。而在大豆根的R(-)Na(+)与R(+)Na(+)的比较中,根瘤菌对上述基因编码的表达调控都呈现显著的下调趋势。特别是大豆根中的MYB-1、CHI-2编码基因的下调趋势最为显著,分别达到了99。8%和98。1%。这表明,根瘤菌通过大豆叶中MYB、CHS、CHI、CPM编码基因增强植物的耐盐胁迫能力,而大豆根中的MYB-1、CHI-2等基因起负调控作用。74231
Abstract Rhizobium could not only improve the soybean nutritional quantity, but also enhance the soybean’s salt tolerance。 However, the molecular mechanisms of these two functions are not clear。 In this study, we analysis the transcriptional expression level of some salt responsive genes, which are also key regulatory genes in the flavonoid metabolic pathway, by qRT-PCR。 Results showed that the transcription factors MYB, CHS, CHI and CPM encoding genes in leaves of Union85140 salt sensitive soybean were significantly up-regulated by the application of the rhizobium, especially the upward trend of the CHI-1 encoding gene was the most significant, reached 64705。8%, and next is CHS-3 encoding gene, reached 6065。8%。 In comparison with R (-) Na (+) and R (+) Na (+) in soybean roots, the expression of the genes coding for above-mentioned genes showed a significant downward trend。 Especially the MYB-1, CHI-2 encoding gene in soybean roots is the most significant downward trend, reached 99。8% and 98。1%。 This suggests that the rhizobium pass through MYB, CHS, CHI and CPM encoding gene in soybean leaves can enhanced plant salt tolerance,and MYB-1, CHI-2 gene can play a negulatory role in soybean root。
毕业论文关键词:大豆; 根瘤菌; 耐盐基因; 实时荧光定量PCR; 转录表达
Key words: soybean; rhizobium; salt tolerance gene; Real time quantitative PCR; transcriptional expression
目录
引言 4
1 材料与方法 5
1。1 大豆的栽培 5
1。1。1 大豆种子的灭菌处理 5
1。1。2 大豆种子的春化、萌发 5
1。1。3 大豆种子的种植 5
1。1。4 盐胁迫处理、收样 5
1。2 大豆植株总RNA的提取 5
1。2。1 材料、仪器设备及试剂 5
1。2。2 试验方法 6
1。3 大豆植株总RNA反转录cDNA 7
1。3。1 材料、仪器设备及试剂 7
1。3。2 试验方法 7
1。4 荧光定量PCR 8
1。4。1 材料、仪器设备及试剂 8
1。4。2 试验方法 8
1。5 数据处理与分析 10
2 结果与分析 10
2。1 根瘤菌施用对盐胁迫环境中MYB盐响应基因转录表达的影响。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。10