发展套作大豆, 振兴大豆产业

目的对大豆阴蔽响应基因GmIF-3进行克隆与表达分析。方法前期采用蛋白磷酸化组学分析了大豆全蛋白组在白光及阴蔽胁迫下的磷酸化水平，筛选到1个受阴蔽诱导磷酸化水平显著升高的IF-3 (translation initiation factor)蛋白，将其基因命名为GmIF-3 (Glyma.17G072300)。利用生物信息学分析其启动子序列、蛋白结构、蛋白疏水性等生化特性；采用Gateway^®克隆方法克隆该基因的cDNA序列；采用实时荧光定量PCR检测该基因的组织表达模式及阴蔽处理下的表达情况。结果生物信息学分析表明：GmIF-3位于大豆第17号染色体上，可编码274个氨基酸残基的蛋白质，其蛋白等电点为9.11，分子量为30.83 ku，启动子含有多个与光信号响应和激素响应的顺式作用元件。实时荧光定量PCR显示：GmIF-3基因在真叶中的相对表达量最高，在三出复叶、茎尖分生组织、根、茎、叶柄、子叶、花和荚等多种器官中也均有表达。对大豆幼苗阴蔽处理0、1、2、4和6 h后，GmIF-3的表达量显著升高，阴蔽6 h的相对表达量最高，约为对照(阴蔽0 h)的8.3倍。结论GmIF-3 基因可能参与大豆对阴蔽胁迫的应答过程。

Legume-based cropping systems have reduced carbon and nitrogen losses

PurposeCloning and expression analysis of shade responsive gene GmIF-3 in soybean.MethodPhosphoproteomics was used to analyze the phosphorylation status of soybean total proteins under the shading stress. We screened out an IF-3 (translation initiation factor) protein whose phosphorylation was significantly elevated in the shade and its encoding gene was named as GmIF-3 (Glyma.17G072300). Bioinformatics was used to analyze its gene structure and biochemical properties such as promoter sequence, protein structure and protein hydrophobicity. The cDNA sequence was cloned by Gateway^® cloning. Real-time quantitative PCR was used to detect its tissue expression pattern and expression under shading treatment.ResultsBioinformatics analysis showed that GmIF-3 is located on soybean chromosome 17, which encodes a protein of 274 amino acid residues with a protein isoelectric point of 9.11 and a molecular weight of 30.83 ku. Its promoter contains multiple light and hormonal response cis-acting elements. The cDNA sequence was then amplified by RT-PCR. Real-time quantitative PCR showed that the relative expression of GmIF-3 gene was the highest in unifoliolate leaves. It is also expressed in various organs such as trifoliolate leaves, shoot apical meristems, roots, stems, petioles, cotyledons, flowers and pods. The expression of GmIF-3 in the seedlings was significantly upregulated as time increase from 0 to 1, 2, 4 and 6 h under shade stress, and reach the highest level at the 6^th h, which is about 8.3 times higher than that in the control (at 0 h).ConclusionThe GmIF-3 gene may be involved in the response of soybean to shading stress.