转录因子BvM14-GAI耐盐功能研究

doi:10.11924/j.issn.1000-6850.casb2021-0696

摘要/Abstract

摘要：

转录因子在植物应答逆境胁迫过程中发挥着重要作用,GAI蛋白是植物转录因子家族的重要一员,对其研究主要集中在光响应机制领域,而该蛋白响应耐盐机制研究报道较少。实验室前期已经获得甜菜 M14品系盐胁迫转录组中上调表达基因BvM14-GAI的cDNA全长,本研究试图阐明该基因参与盐胁迫的功能。通过构建该基因植物表达载体,利用农杆菌介导花序浸染法转化拟南芥野生型和GAI基因突变株,检测150 mmol/L NaCl胁迫下异源表达和异源互补拟南芥植株的表型和生理生化指标。0 mmol/L NaCl处理时,以拟南芥野生型和GAI基因突变株为对照,异源表达和异源互补植株的根长、鲜重和干重均显著低于对照,说明BvM14-GAI基因为生长负调控因子;150 mmol/L NaCl胁迫处理后的根长、鲜重、干重及K⁺/Na⁺差异不显著,但甜菜碱、SOD和POD酶活性的含量显著增加,表明转录因子BvM14-GAI通过增强渗透调节和抗氧化酶系统提高异源表达和异源互补拟南芥植株的耐盐功能。研究结果不仅拓展了植物GAI基因响应非生物胁迫的功能,而且对阐明甜菜M14品系耐盐分子机制和培育耐盐作物品系具有一定研究价值。

关键词: 甜菜M14品系, BvM14-GAI基因, 盐胁迫, 转录因子, 抗氧化酶

Abstract:

Transcription factors play an important role in the process of plant response to adversity stress. GAI protein is an important member of the plant transcription factor family. About this protein, the research mainly focuses on the field of light response mechanism, while the protein response to salt tolerance mechanism is less reported. The full-length cDNA of BvM14-GAI gene, which is up-regulated in the transcriptome of the sugar beet M14 line under salt stress, is obtained in the early stage of the laboratory. This study aims to clarify the function of this gene in salt stress. By constructing the plant expression vector of BvM14-GAI gene, it is transformed into the Arabidopsis wild-type and GAI gene mutant plants by Agrobacterium-mediated inflorescence impregnation, then the phenotype and the indexes of the physiology and biochemistry of heterologous expression and heterologous complementary Arabidopsis plants under 150 mmol/L NaCl stress are detected. When treated with 0 mol/L NaCl, the root length, fresh weight and dry weight of heterologous expression and heterologous complementary Arabidopsis plants are significantly lower than those of the control group, indicating that BvM14-GAI gene is a negative growth regulator. After 150 mmol/L NaCl stress treatment, there is no significant difference in root length, fresh weight, dry weight and K ⁺/Na⁺ of the transgenic plants, but the contents of betaine, the enzyme activities of SOD and POD of the transgenic plants increase significantly, indicating that the transcription factor BvM14-GAI could improve the salt tolerance function of heterologous expression and heterologous complementation Arabidopsis plants by enhancing osmotic regulation and antioxidant enzyme systems. The study not only expands the function of plant GAI gene in response to abiotic stress, but also has research value for elucidating the salt-tolerant molecular mechanism of sugar beet M14 line and cultivating salt-tolerant crop lines.

Key words: sugar beet M14 line, BvM14-GAI gene, salt stress, transcription factor, antioxidant enzyme

中图分类号:

Q785

马慧敏, 孙培琳, 马春泉. 转录因子BvM14-GAI耐盐功能研究[J]. 中国农学通报, 2021, 37(34): 34-42.

Ma Huimin, Sun Peilin, Ma Chunquan. Salt Tolerance Function of Transcription Factor BvM14-GAI[J]. Chinese Agricultural Science Bulletin, 2021, 37(34): 34-42.

图/表 9

图1. 35S::pCAMBIA1300-BvM14-GAI重组质粒特异引物PCR产物1%琼脂糖凝胶电泳图 M为DNA Marker DL 2000,1~5为pCAMBIA1300-BvM14-GAI重组质粒特异引物PCR产物

图2. 拟南芥GAI基因突变株纯合子筛选PCR筛选结果 M为DNA Marker DL 2000,1~12为拟南芥GAI基因突变株,13为拟南芥野生型

图3. BvM14-GAI基因异源表达及异源互补阳性拟南芥植株的抗性筛选(Hyg 30 mg/L)结果

图4. BvM14-GAI基因拟南芥异源表达植株及异源互补植株的PCR检测结果 M为DNA Marker DL 2000,WT为野生型植株,OX为BvM14-GAI基因异源表达植株,KO为GAI基因突变株,CO为BvM14-GAI基因异源互补植株,下同

图5. BvM14-GAI基因拟南芥异源表达植株及异源互补植株的RT-PCR检测结果

图6. BvM14-GAI基因拟南芥异源表达及异源互补植株盐胁迫前后的表型及根长测定结果不同小写字母代表显著性差异(P<0.05),下同

图7. BvM14-GAI基因拟南芥异源表达及异源互补植株盐胁迫前后的鲜重、干重测定结果

图8. BvM14-GAI基因拟南芥异源表达及异源互补植株盐胁迫前后K+/Na+及甜菜碱含量测定结果

图9. BvM14-GAI基因拟南芥异源表达及异源互补植株盐胁迫前后SOD、POD活性测定结果

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