• 中文核心期刊
  • CSCD来源期刊
  • 中国科技核心期刊
  • CA、CABI、ZR收录期刊

水稻优良三系不育系福农A多抗性基因分析及表达特性研究

Disease and Insect Resistance Genes in Premium Sterile Line Funong A

    摘要
  • 摘要:
    目的 分析福农A中的抗病虫基因，为该不育系更好地应用于育种生产提供理论依据。
    方法 以福农A及亲本福稻B、华航丝苗为材料，利用抗稻瘟病基因分子标记检测各材料中抗稻瘟病基因情况，并应用高密度芯片检测抗白叶枯病、抗病毒及抗褐飞虱基因情况。PCR扩增获得福农A及亲本福稻B、华航丝苗中的抗稻瘟病基因，进行序列比对分析。植株培养至三叶一心期，喷雾接稻瘟病菌并取样，采用 SYBR Green I 荧光定量 PCR（qRT-PCR） 分析抗稻瘟病基因的表达模式。
    结果 福农A及亲本福稻B、华航丝苗中均含有稻瘟病抗性基因Pib、Pia、Pid3和Pid2，但均不含Pi9、Pi54、Pigm、Pit、Pi2；另外，福农A和华航丝苗中含有Pi5，而福稻B中含有Pita和Pi37。福农A及亲本福稻B和华航丝苗中均含有抗白叶枯病基因Xa21和抗黄色斑驳病毒病基因Rymv1；福农A及华航丝苗中含持久性抗水稻条叶枯病毒基因STV11；但它们中均不含抗褐飞虱基因Bph14、Bph15、Bph18、 Bph26、 Bph6、 Bph9。扩增获得的 Pi5、Pia、Pib、Pid3、Pid2基因序列长分别为5 672、2 597、5 532、2 865、3847 bp；福农A与亲本华航丝苗中的Pi5基因序列完全一致；福农A和亲本华航丝苗、福稻B中的Pia、Pib基因序列均完全一致；福农A和华航丝苗中的Pid3和Pid2基因序列完全一致，而它们与福稻B的序列分别有2个和5个碱基差异。在福农A中，抗稻瘟病基因Pi5和Pib的表达明显受稻瘟病菌的诱导，接菌72 h时表达量最高；Pia和Pid3的表达随接菌时间的延长而逐渐升高，接菌96 h时表达量最高；Pid2的表达先升高后降低，且在接菌24 h时表达量最高。
    结论 水稻三系不育系福农A中含有稻瘟病抗性基因Pi5、Pib、Pia、Pid3和Pid2，抗白叶枯病基因Xa21、抗黄色斑驳病毒病基因Rymv1及持久性抗水稻条叶枯病毒基因STV11。因此，该不育系有望应用于多抗基因的聚合育种研究。

     

    Abstract:
    Objective Disease and insect resistance genes in Funong A, a three-line indica cytoplasmic male sterile line, were analyzed for breeding applications.
    Method  Genomic DNA of Funong A, Fudao B, and Huahangsimiao rice germplasms was extracted by CTAB method followed by detection of blast resistance genes with molecular markers. Resistance genes related to bacterial blight, viral diseases, and brown planthopper infestation were analyzed by high density chip detection method. Blast resistance genes of the lines were obtained by PCR and sequenced for comparison. Specimens were collected at 0, 24, 48, 72, and 96 h after a blast fungus inoculation for extraction of total RNA using Trizol method. Gene expression was determined by SYBR Green I qRT-PCR.
    Result Funong A, Fudao B, and Huahangsimiao were found to contain the blast resistance genes Pib, Pia, Pid3, Pid2, bacterial blight resistance gene Xa21, and yellow mottle virus resistance gene Rymv1. Furthermore, Funong A and Huahangsimiao had the blast resistance gene Pi5 and the stripe virus resistance gene STV11, while Fudao B consisted of the blast resistance genes Pita and Pi37. However, no brown planthopper resistance genes were detected in them. The fragments of Pi5, Pia, Pib, Pid3, and Pid2 were 5 672 bp, 2 597 bp, 5 532 bp, 2 865 bp, and 3 847 bp in length, respectively. The sequences of Pi5, Pid3, and Pid2 in Funong A were similar to those in Huahangsimiao, but 2 bases differed from Pid3 and 5 from Pid2 of Fudao B. In Funong A, the expressions of rice blast resistance genes Pi5 and Pib were significantly induced by the blast fungus with a peak occurred 72 h after inoculation. The expressions of Pia and Pid3 increased gradually with time after inoculation to peak in 96 h. The expression of Pid2 rose initially to a maximum in 24 h and then declined.
    Conclusion The three-line indica cytoplasmic male sterile line Funong A carried the blast resistance genes Pi5, Pib, Pia, Pid3, and Pid2, which were significantly induced by the blast fungus. The line also contained the bacterial blight resistance gene Xa21, yellow mottle virus resistance gene Rymv1, and stripe virus resistance gene STV11 but none of the insect resistance genes.

     

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