CHEN Yan-ning,WU Zhi-yi,YUAN Wen-jie,et al.Research on the Regulation Effect of Ribosomal Gene GmRPL12 on Low Sulfur Tolerance in Soybean[J].Soybean Science,2020,39(04):518-526.[doi:10.11861/j.issn.1000-9841.2020.04.0518]
核糖体基因GmRPL12对大豆低硫耐性的调控作用研究
- Title:
- Research on the Regulation Effect of Ribosomal Gene GmRPL12 on Low Sulfur Tolerance in Soybean
- 文献标志码:
- A
- 摘要:
- 为研究大豆中核糖体基因对大豆耐低硫胁迫功能的调控作用，从科丰1号根中克隆1个大豆核糖体蛋白编码基因GmRPL12。分析基因结构和低硫胁迫下不同组织中的表达情况，将基因过表达载体和干扰载体转化科丰1号毛状根,获得转基因嵌合体,并进行基因表达分析和植株表型分析。序列分析结果表明：基因编码区全长576 bp，预测蛋白C端包含1个核糖体蛋白L7/L12保守结构域RPL12。该基因在根中表达量较高，表达水平受低硫诱导，不同品种中该基因对低硫响应的模式不同。通过毛状根遗传转化获得GmRPL12基因过表达、RNA干扰以及空载体对照的大豆嵌合体。低硫处理与正常硫水平处理相比，GmRPL12基因过表达的大豆嵌合体植株倒一叶和倒二叶SPAD、株高、地上部鲜重和干重、地下部鲜重和干重显著增加，RNA干扰下调大豆嵌合体植株的以上指标。GmRPL12基因的过量表达能显著增加低硫处理时地上部、地下部的无机硫含量，以及正常硫水平处理时地上部的无机硫含量。研究结果暗示GmRPL12基因可能参与大豆对低硫耐性和大豆硫代谢的调控。
- Abstract:
- In order to investigate the regulation of ribosomal genes on soybean tolerance to low sulfur stress, this study cloned a soybean ribosomal gene GmRPL12 from the root of Kefeng 1. We analysised the gene structure and expression in different tissues under low sulfur stress.The gene overexpression vector and interference vector were transformed into hairy root of Kefeng 1 to obtain transgenic chimeras, and then we analysised the expression of the gene and the phenotype of the chimera plant.The results of sequence analysis showed that, the total length of the gene code region was 576 bp. There was a ribosomal protein L7/L12 C-terminal domain RPL12 in its predicted protein. The gene was highly expressed in roots and was induced by low sulfur, and showed different expression patterns in two materials. Soybean chimeras with gene overexpression (OE), RNA interference (Ri), and two empty vectors (OE-EV and Ri-EV) were obtained by genetic transformation of hairy roots. Compared with that under the +S condition, SPAD value, the plant height, the fresh weight and dry weight of aboveground, the fresh weight and dry weight of root increased significantly in the GmRPL12 gene-overexpressed soybean chimera under the -S condition, and these indicators decreased significantly in the soybean Ri chimera under the -S condition. Furthermore, the inorganic sulfur contents of soybean OE chimeras increased significantly in the above ground and under ground plant under -S, and in the above ground plant under +S. These results suggest that GmRPL12 gene may be involved in soybean tolerance to low sulfur and soybean sulfur metabolism.
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