Molecular marker for bacterial stripe resisting major gene BLS1 locus of rice and application of molecular marker

A technology of major genes and molecular markers, applied in the field of molecular biology, can solve the problems of in-depth molecular basic research and less research on molecular genetic mechanism, and achieve the effects of saving screening workload, clear location, and high cost

Active Publication Date: 2016-10-12
广西壮族自治区农业科学院水稻研究所
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  • Abstract
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  • Application Information

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Problems solved by technology

Most of the studies on the main effect genes of resistance to thin stripe disease are concentrated on the screening of resistance sources and the analysis of resistance genetics, while t

Method used

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  • Molecular marker for bacterial stripe resisting major gene BLS1 locus of rice and application of molecular marker
  • Molecular marker for bacterial stripe resisting major gene BLS1 locus of rice and application of molecular marker
  • Molecular marker for bacterial stripe resisting major gene BLS1 locus of rice and application of molecular marker

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Experimental program
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Effect test

Embodiment 1

[0043] 1. The origin of the main gene BLS1:

[0044] Huang Dahui et al. (2008) first found that common wild rice DP3 is resistant to bacterial streak type III by using resistance identification screening (Huang Dahui, Cen Zhenlu, Liu Chi, He Wenai, Chen Yingzhi, Ma Zengfeng, Yang Lang, Wei Shaoli, Liu Yali, Huang Siliang, Yang Xinqing, Li Rongbai. Screening and genetic analysis of wild rice bacterial streak resistance resources. Journal of Plant Genetic Resources, 2008, 9(1):11-14). He et al. (2012) showed through genetic analysis that DP3 carries a pair of recessive bacterial stripe disease resistance genes and temporarily named it bls1, and located the gene in the 4.0-cM region between RM587 and RM510 (HeW A, Huang D H*,Li R B,Qiu Y F,Song J D,Yang H N,Zheng J X,Huang Y Y,Li X Q,Liu C,Zhang Y X,Ma Z F and Yan Y. Identification of a resistance gene bls1to bacterial leaf streak in wild rice Oryza rufipogon Griff . Journal of Integrative Agriculture, 11(6):962-969). This gene...

Embodiment 2

[0060] Example 2: Verification of Molecular Markers

[0061] 1. Materials and methods

[0062] 1.1 Materials

[0063] Negative varieties: 10 copies, 8 copies of non-insect-resistant materials in the breeding combination of susceptible variety 93-11, resistant parent DP3, and DP3X 93-11.

[0064] Positive varieties: 9 insect-resistant materials in the breeding combination of insect-resistant varieties DP3 and 93-11X DP3.

[0065] Molecular marker primers: RM19382, RM 19391, RM19400, RM 19402, RM510.

[0066] 1.2 Method

[0067] Genomic DNA of rice sample was extracted by CTAB extraction method (method is the same as in Example 1). Sample DNA was amplified with primers RM19382, RM19391, RM19400, RM19402 and RM510, respectively. The reaction system included 0.10 μM primers, 250 μM dNTPs, 1× PCR reaction buffer (50 mM KCl, 10 mM Tris-HCl pH8.3, 1.5 mM MgCl 2 ), 100ng of DNA template, 1UTaq enzyme. The reaction program was: pre-denaturation at 94°C for 5 minutes, cycle (94°C...

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Abstract

The invention discloses a molecular marker for a bacterial stripe resisting major gene BLS1 locus of rice and an application of the molecular marker. Specific steps for screening are as follows: (1) constructing a positioned segregation population, so as to obtain an approximate isogenic line F2 of the positioned segregation population; (2) extracting genomic DNA of rice leaves of each single strain of parents and a F2 population by adopting a CTAB method so as to carry out SSR molecular marker analysis; (3) preliminarily positioning a gene BLS1 in a region between RM19382 and RM510; (4) carrying out BLS1 close-linkage marking: carrying out detection and analysis after carrying out molecular marking by several kinds of marking primers, so as to position the BLS1 in a physical range, i.e., 21-kb between RM19400 and RM510. The molecular marker is applied to the selective breeding of bacterial stripe resisting rice varieties or the screening of resistant genetic resources. The molecular marker disclosed by the invention can be used for effectively detecting whether ordinary bacterial stripe resisting wild rice DP3 and derived varieties (lines) thereof contain the major gene locus or not, the efficiency of selection of bacterial stripe resisting rice is increased, and the bacterial stripe resisting rice varieties containing the gene BLS1 are obtained.

Description

【Technical field】 [0001] The invention relates to the field of molecular biology, in particular to a molecular marker for the BLS1 locus of the rice anti-bacterial leaf spot main gene. 【Background technique】 [0002] Bacterial streak disease (referred to as thin stripe disease) is caused by the pathogen Xanthomonas oryze pv. Oryzicola (Xoc), and is an important rice disease. The disease can cause a 32% loss of rice yield in an outbreak year. Cultivation measures and the use of pesticides are effective ways to prevent and control thin stripe disease. However, using resistant resources to breed resistant varieties is the most economical and effective way to control the disease. Genetic analysis showed that there were two different types of resistance resources, such as major genes and quantitative trait loci (Quantitative trait locus, QTL). At present, there are few researches on the application of molecular markers of the major genes of anti-bacterial stripe disease. [0...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
Inventor 黄大辉秦钢马增凤刘驰罗同平张月雄岑贞陆
Owner 广西壮族自治区农业科学院水稻研究所
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