YANG Jing,ZHAO Qian-qian,NIU Lu,et al.Study on the Enhancement of Soybean Resistance to Sclerotinia Sclerotiorum with Transformation of Cmoxdc1 Gene from Coniothyrium Minitans[J].Soybean Science,2020,39(05):712-719.[doi:10.11861/j.issn.1000-9841.2020.05.0712]
转盾壳霉Cmoxdc1基因增强大豆对菌核病抗性的研究
- Title:
- Study on the Enhancement of Soybean Resistance to Sclerotinia Sclerotiorum with Transformation of Cmoxdc1 Gene from Coniothyrium Minitans
- Keywords:
- Soybean; Sclerotinia stem rot; Sclerotinia sclerotiorum; Oxalic acid; Cmoxdc1; Resistance
- 文献标志码:
- A
- 摘要:
- 草酸被认为是核盘菌的主要致病因子，在病原菌的侵染过程中发挥着重要作用，并与核盘菌的致病力密切相关，为研究转盾壳霉草酸脱酸酶基因Cmoxdc1对大豆菌核病抗性的促进作用，本研究利用农杆菌介导转化法，将来源于核盘菌生防菌盾壳霉（Coniothyrium minitans）的Cmoxdc1基因导入大豆感病品种Jack，研究转Cmoxdc1基因大豆对草酸耐受性和菌核病抗性。结果表明：外源基因以低拷贝形式（1～2个）整合至大豆基因组，并能够完成转录。对离体叶片进行60 mmol?L-1草酸处理后，转基因植株叶片病斑面积显著低于对照受体品种，接种7 d后病斑面积较受体对照减少88.80%～89.98%。离体叶柄和叶片接种核盘菌后，转基因大豆病斑扩展受到明显抑制，病斑长度和病斑面积均显著低于对照受体。接种4 h后，叶柄病斑长度为对照品种的41.38%～43.51%；接种7 d后，叶片病斑面积为对照品种的8.68%～27.01%；并且，接种核盘菌后转基因叶片中的草酸含量也较对照品种显著降低。研究结果表明盾壳霉Cmoxdc1表达显著增强转基因大豆对草酸的耐受性和对菌核病的抗性。
- Abstract:
- It is widely accepted that oxalic acid (OA) is a key virulence factor which plays an essential role in the pathogenesis of sclerotinia stem rot (SSR) and is closely related to the pathogenicity of S.sclerotiorum. In this study, an oxalic acid decarboxylase gene Cmoxdc1 from Coniothyrium minitans, a ubiquitous antagonistic fungus of S.sclerotiorum, was introduced into the soybean cultivar Jack susceptible to SSR, and stable transgenic lines were generated to evaluate for their tolerance to OA and resistance to SSR. Molecular characterization confirmed the integration with single or double copies of insertions and expression at the transcriptional level of the foreign gene Cmoxdc1 in transgenic soybean. OA treatment with externally applied OA (60 mmol?L-1) showed that the transgenic plants exhibited significant reduced lesion size on the detached leaves by 88.80%-89.98% reduction of lesion area relative to the non-transformed (NT) plants at 7 d after OA application. S.sclerotiorum inoculation revealed limited lesion development and less lesion sizes in transgenic plants, with 41.38%-43.51% of lesion lengths (4 d after inoculation) in the detached stem assay and 8.68%-26.98% of lesion areas (7 d after inoculation) in the detached leaf assay, respectively, as that of the corresponding NT control. Meanwhile, decreased OA content were also observed in the transgenic leaves after S.sclerotiorum inoculation. In conclusion, the results demonstrate that the expression of Cmoxdc1 from C.minitans significantly enhanced OA tolerance and SSR resistance in transgenic soybean plants.
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