Two novel gene-specific markers at the Pik locus facilitate the application of rice blast resistant alleles in breeding

doi:10.1016/S2095-3119(20)63272-5

Journal of Integrative Agriculture ›› 2021, Vol. 20 ›› Issue (6): 1554-1562.DOI: 10.1016/S2095-3119(20)63272-5

所属专题：植物抗病遗传合辑Plant Disease-resistance Genetics

收稿日期:2020-02-17 出版日期:2021-06-01 发布日期:2021-05-19

Two novel gene-specific markers at the Pik locus facilitate the application of rice blast resistant alleles in breeding

TIAN Da-gang^{1, 2}, CHEN Zi-qiang¹, LIN Yan¹, CHEN Zai-jie¹, LUO Jia-mi¹, JI Ping-sheng⁴, YANG Li-ming³, WANG Zong-hua², WANG Feng¹

¹ Fujian Key Laboratory of Genetic Engineering for Agriculture/Biotechnology Research Institute, Fujian Academy of Agricultural Sciences, Fuzhou 350003, P.R.China
² State Key Laboratory of Ecological Pest Control for Fujian and Taiwan Crops, College of Life Science, Fujian Agriculture and Forestry University, Fuzhou 350002, P.R.China
³ College of Biology and the Environment, Nanjing Forestry University, Nanjing 210037, P.R.China
⁴ Department of Plant Pathology, University of Georgia, Tifton, GA 31793, USA

Received:2020-02-17 Online:2021-06-01 Published:2021-05-19
Contact: Correspondence YANG Li-ming, E-mail: yangliming@njfu.edu.cn; WANG Zong-hua, E-mail: wangzh@fafu.edu.cn; WANG Feng, E-mail: wf@fjage.org
Supported by:
This research was funded by the National Natural Science Foundation of China (31640006), the China Postdocotral Science Foundation (2019M662219), the Youngth Program of Fujian Academy of Agricultural Sciences (YC2019004), and the Projection of Public Welfare of Fujian Province (2017R1019-10).

摘要/Abstract

摘要：

由稻瘟病菌引起的稻瘟病，是一种制约世界水稻生产的真菌病害。长期的生产实践表明，将持久广谱的抗性基因导入高产水稻品种，是防治该病害的首选。位于第11号染色体上的抗瘟基因 Pik 基因座，至少含有 Pi-1、Pik-h、Pi-k、Pik-m、Pik-s和Pik-p等6个重要的抗病基因；但由于当前缺乏适用的分子标记，限制了该基因座功能基因在抗病育种中的广泛应用。为了更好地在分子育种中利用该基因座的功能基因，开发Pik 基因座功能基因的特异性标记并用其分型种质资源具有重要意义。基于此，本研究通过对Pi-1、Pik-h、Pi-k、Pik-m、Pik-s、Pik-p等功能基因和非功能基因位点之间的序列广泛比较，获得了一个在这些功能基因启动子区-1015-bp处Pik-p缺失19-bp和一个在这些功能基因末端+6816-bp处Pi-1插入11-bp的的多态性位点，并据此分别开发出两个能精确区分出Pik-p、Pi-1和K型功能等位位点的Pikp-Del和Pi1-In标记；进一步通过结合稻瘟病菌室内接种和已有的dCAPS标记Pi1FNP和dCAPS-795鉴定，对这两个标记鉴定结果的准确性进行了评价。结果显示，我们鉴定的基因型与稻瘟病人工接种抗性表现和两个dCAPS标记的鉴定的结果完全一致。另外，我们还利用Pikp-Del和Pi1-In标记对531份水稻品种和育种材料进行了基因分型，结果表明，5份材料含有Pik-p基因，8份材料含有Pi-1基因，说明这两个基因在我国水稻稻瘟病抗性育种中还没有被充分利用；另外还有256份携带K型等位基因，这些材料可作为抗稻瘟病育种的种质资源。综上，Pikp-Del和Pi1-In标记可以实现对Pik-p、Pi-1和K型功能等位基因的精准检测，结合标记分型的种质资源，将会加速Pik-p 和Pi-1以及Pik 基因座的其它功能基因在抗病育种中的应用。

Abstract:

Blast, a disease caused by Magnaporthe oryzae, is a major constraint for rice production worldwide. Introgression of durable blast resistance genes into high-yielding rice cultivars has been considered a priority to control the disease. The blast resistance Pik locus, located on chromosome 11, contains at least six important resistance genes, but these genes have not been widely employed in resistance breeding since existing markers hardly satisfy current breeding needs due to their limited scope of application. In this study, two PCR-based markers, Pikp-Del and Pi1-In, were developed to target the specific InDel (insertion/deletion) of the Pik-p and Pi-1 genes, respectively. The two markers precisely distinguished Pik-p, Pi-1, and the K-type alleles at the Pik locus, which is a necessary element for functional genes from rice varieties. Results also revealed that only several old varieties contain the two genes, of which nearly half carry the K-type alleles. Therefore, these identified varieties can serve as new gene sources for developing blast resistant rice. The two newly developed markers will be highly useful for the use of Pik-p, Pi-1 and other resistance genes at the Pik locus in marker-assisted selection (MAS) breeding programs.

Key words: rice , blast disease , molecular marker , Pik-p , Pi-1 , K-type alleles

. [J]. Journal of Integrative Agriculture, 2021, 20(6): 1554-1562.

TIAN Da-gang, CHEN Zi-qiang, LIN Yan, CHEN Zai-jie, LUO Jia-mi, JI Ping-sheng, YANG Li-ming, WANG Zong-hua, WANG Feng . Two novel gene-specific markers at the Pik locus facilitate the application of rice blast resistant alleles in breeding[J]. Journal of Integrative Agriculture, 2021, 20(6): 1554-1562.

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Two novel gene-specific markers at the Pik locus facilitate the application of rice blast resistant alleles in breeding

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