中国农业科学

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黑龙江省水稻种质抗瘟评价及抗瘟遗传结构的分析

张亚玲,高清,赵羽涵,刘瑞,付忠举,李雪,孙宇佳,靳学慧   

  1. 黑龙江八一农垦大学农学院/黑龙江省植物抗性研究中心,黑龙江大庆 163000
  • 发布日期:2021-12-19

Evaluation of blast resistance and genetic structure analysis of rice germplasm in Heilongjiang Province

ZHANG YaLing, GAO Qing, ZHAO Yuhan, LIU Rui, FU Zhongju, LI Xue, SUN Yujia, JIN XueHui   

  1. College of Agronomy, Heilongjiang Bayi Agricultural University/Heilongjiang Plant Resistance Research Center, Daqing 163000, Heilongjiang
  • Online:2021-12-19

摘要: 【目的】稻瘟病严重威胁黑龙江省水稻的生产,选育和利用抗瘟品种是最经济、安全和有效措施。了解黑龙江省水稻主栽品种的稻瘟病抗性,明确抗病基因在黑龙江省抗性效应,为黑龙江省稻瘟病抗病种质资源的选育和利用提供依据。【方法】2018年秋季采自黑龙江省水稻主产区的134个水稻单孢菌株,采用病原物接种鉴定方法对黑龙江省50个水稻主栽品种进行抗性分析针对已报35个稻瘟病抗病基因,利用基因特异引物对参试品种及阳性对照品种进行抗瘟基因的检测,部分无阳性对照品种抗性基因扩增后测序,与NCBI中公布的参考序列进行比对,分析稻瘟病抗性基因在相应品种中存在情况通过基因聚合类型与品种抗性表现相关性分析,明确与黑龙江省水稻品种抗性表现相关的基因型。【结果】根据抗性频率评价,参试的黑龙江省50份水稻品种中龙粳20表现为抗,龙粳67、龙垦202、龙粳40、龙粳31、龙粳57和龙粳43表现为中抗,其余43个品种均表现感病。通过品种组合抗性分析发现龙粳20龙粳6733对品种组合的联合抗性系数(association coefficients,RAC)较高,联合致病性系数(virulence association coefficientsVAC较低,联合抗病性较好,该类搭配结构具有良好应用前景。通过特异性引物对品种的携带的抗瘟基因鉴定结果显示,抗瘟基因PishPi36Pi33Pi-CO39在供试品种中均被检出,Pi63PtrPi37Pi64pi21Pi9Pi54Pi-khPiaPikpPi35PikmPik检出率为50%—100%,说明这类基因在黑龙江省水稻育种中应用较广泛PitaPibPiiPi5Piz-tPi50Pi2检出率为10%—50%Pid2仅在2个品种中被检出,Pigm仅在吉粳88中被检出,而PitPid3Bsr-d1Pi25Pid3-A4Pi56Pi1PikePb1在供试品种中均未检测到,说明这类基因在黑龙江省水稻品种中分布较少。通过品种基因型分析发现,供试品种携带抗瘟基因12—19个不等,共58种抗瘟基因型,说明供试品种的抗瘟基因组合类型较丰富。单基因及多基因聚合与抗病相关性分析结果显示,Pi2Piz-tPi50Pi5Pii的分布频率与携带该基因且表现抗病的百分率相当;研究发现品种携带抗瘟基因越多其表现为抗病品种的概率越高,携带Pi2+Piz-t+Pi50+α聚合类型的6份品种均表现抗病。【结论】参试的黑龙江省水种质资源抗性偏低,部分品种组合种植有较高应用价值,抗瘟基因在参试品种中分布不等,Pi2Piz-tPi50Pi5Pii在品种抗病方面起主导作用Pi2+Piz-t+Pi50+α基因聚合类型可以提高水稻抗瘟性


关键词: 水稻, 稻瘟病, 抗性频率, 抗瘟性评价, 抗瘟基因

Abstract: ObjectiveRice blast seriously threatens rice production in Heilongjiang Province and the breeding and utilization of anti-blast varieties are the most economical, safe and effective measures to control it. This study explored the resistance of rice varieties in Heilongjiang Province; clarified the resistance effect of disease resistance genes, and provided a basis for the selection and utilization of germplasm resources of rice blast in Heilongjiang Province. MethodIn the autumn of 2018,134 monospore rice strains were collected from the main rice area in Heilongjiang Province, and the resistance of 50 dominant rice varieties was analyzed. For the reported 35 rice blast resistance genes, a comparison was made between the varieties and positive control varieties. Sequencing results of some positive control varieties were compared with the reference sequence published in the National Centre for Biotechnology Informationhttps://www.ncbi.nlm.nih.gov/ to analyze the existence of rice blast resistance genes in the corresponding varieties. Through the correlation analysis of gene aggregation type and breed resistance performance, the genotypes related to the resistance performance of rice varieties in Heilongjiang Province were clarified. ResultsAmong 50 rice varieties in Heilongjiang Province, Longgeng 20 had the best resistance (R). Longgeng 67, Longken 202, Longgeng 40, Longgeng 31, Longgeng 57 and Longgeng 43 had moderate disease resistance; whereas, 43 varieties such as Jiahe 1 had relatively low resistance. Through the analysis of variety combination resistance, we found that 33 pairs of Longgeng 20 + Longgeng 67 had a high RACresistance association coefficient) value, low VAC (virulence association coefficient) value and good combined disease resistance. The matching structure showed potential for application. The identification of resistance genes carried by specific primers showed that Pish, Pi36, Pi33 and Pi-CO39 were detected, and Pi63, Ptr, Pi37, Pi64, pi21, Pi9, Pi54, Pikh, Pia, Pikp, Pi35, Pikm and Pik were between 50–100% of occurences frequency, indicating that such genes were widely used in rice breeding in Heilongjiang Province. The detection rate of Pita, Pib, Pii, Pi5, Piz-t, Pi50 and Pi2 was between 10% and 50%; Pid2 was detected in in two varieties, and Pigm was detected in Jigeng 88. However, Pit, Pid3, Bsr-d1, Pi25, Pid3-A4, Pi56, Pi1, Pike and Pb1 were not detected in the cultivars, indicating that such genes are less distributed in rice varieties in Heilongjiang Province. The variety genotype analysis found that the varieties carried 12–19 resistant genes, with a total of 58 genotypes, indicating that the test varieties had rich blast resistance gene combination types. By analysis, the single gene and gene polymerization with disease resistance showed that the distribution frequency of Pi2, Piz-t, Pi50, Pi5 and Pii corresponded to frequency of resistance. We found that the more varieties carrying resistance genes, the higher the frequency resistance, and six varieties that carrying Pi2+Piz-t+Pi50 polymerization types showed resistance. ConclusionOur results have demonstrated the following: the resistance of rice germplasm resources in Heilongjiang province is low; combination planting of different varieties could be usefully applied; the distribution of blast resistance genes is different in the participating varieties; Pi2, Piz-t, Pi50, Pi5 and Pii play a leading role in disease resistance, and Pi2+Piz-t+Pi50+α gene polymerization type could contribute to improving rice blast resistance.


Key words: rice, rice blast, resistance frequency, blast resistance evaluation, blast resistance gene