Rice blast resisting gene Pi-y43 (t) and SSR (simple sequence repeat) markers relative to gene Pi-y43 (t)

A rice and genome technology, applied in genetic engineering, plant gene improvement, DNA/RNA fragments, etc., can solve the problem of distinguishing plants with multiple disease-resistant genes, it is difficult to screen strains containing multiple disease-resistant genes, and resistance Problems such as the inability of diseased strains to distinguish between multiple resistance loci

Inactive Publication Date: 2013-04-24
福建省农业科学院水稻研究所
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  • Application Information

AI Technical Summary

Problems solved by technology

In the traditional method, through the identification of resistance at the seedling stage, it is difficult to screen lines containing multiple disease-resistant genes. Plants containing a single resistance locus have complete resistance and cannot be distinguished from plants containing multiple disease-resistant genes. Results The screened resistant lines could not distinguish whether they contained multiple resistance sites

Method used

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  • Rice blast resisting gene Pi-y43 (t) and SSR (simple sequence repeat) markers relative to gene Pi-y43 (t)
  • Rice blast resisting gene Pi-y43 (t) and SSR (simple sequence repeat) markers relative to gene Pi-y43 (t)
  • Rice blast resisting gene Pi-y43 (t) and SSR (simple sequence repeat) markers relative to gene Pi-y43 (t)

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Experimental program
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Effect test

Embodiment 1

[0020] 1 Mapping population: F hybrid obtained from the broad-spectrum disease-resistant japonica rice variety Yunyin (Yunyin, provided by the Yunnan Academy of Agricultural Sciences) and the common rice blast variety Lijiangxintuanheigu (Lijiangxintuanheigu, LTH). 2 As a fine-tuned population, the population consisted of 312 extremely susceptible plants and 50 disease-resistant plants. Through polymorphic screening of parents Yunyin, Lijiang Xintuan Heigu and Kangganchi, 10 pairs of markers containing polymorphisms were found. The screened polymorphic marker pair F 2 The mapping population was analyzed, wherein 5 recombinants were detected by marker RM3535, and 1 recombinant was detected by RM14166 ( figure 1 ), and two SSR markers, RM6307 and RM208, which were co-segregated with the target gene Pi-y43(t), were detected in the target region. Because this region contains the cloned rice blast resistance gene Pib, the mapping population was further analyzed with the dominant ...

Embodiment example 2

[0033] 1 Gene prediction and sequence analysis of the target gene region

[0034] The sequences of the Pi-y43(t) gene region defined by the markers FJK42 and RM14184 were predicted and analyzed using ORF Finder provided by NCBI, RiceGAAS on the RGP website, FGENESH, GRAMENE, RAPDB and RGAP of Soft berry ( Figure 5 ), Pi-y43(t) was located in the 133kb co-segregation region between the markers FJK42 and RM14184, and there were 1 and 3 recombinants between the two markers, and finally determined 6 candidate genes (as shown in the table below) .

[0035] Candidate genes related to disease resistance in the target region of Pi-y43(t) gene

[0036]

[0037] 2 RT-PCR analysis of candidate genes

[0038] In order to further verify the results of gene prediction, the expression level analysis of candidate genes was carried out, that is, RT-PCR analysis. To this end, RNA was extracted from the leaves of the resistant variety Yunyin and the susceptible varieties Lijiang Xintuan Hei...

Embodiment example 3

[0048] The upstream and downstream primers were designed using the pib DNA sequence registered in NCBI as a template, and the genomic DNA of Yunyin was extracted for PCR amplification ( Figure 9 ), reclaim the PCR product, and connect the PGM-T carrier. After PCR detection of bacterial fluid, positive clones were sent for sequencing. The results showed that the obtained sequence was 5404bp in length and had 100% homology with the pib DNA sequence registered in NCBI.

[0049] Implementation Case 4

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Abstract

The invention provides a rice blast resisting gene Pi-y43 (t) and SSR (simple sequence repeat) markers relative to the gene Pi-y43 (t), which belong to the field of biotechnology, and particularly relate to japonica rice cloud-led rice blast resisting gene Pi-y43 (t) and 5 pairs of SSR pleomorphism markers relative to the gene Pi-y43 (t). The gene is formed on rice blast broad spectrum resistance variety cloud-led secondary chromosome, and is obtained on the basis of fine positioning and map-based cloning of the japonica rice cloud-led paddy anti-rice blast gene Pi-y43 (t). Compared with that with a known resistance gene Pib, a strain containing the rice blast resisting gene Pi-y43 (t) is highly resistant to the cloud-led 'Sichuan-43' rice blast, and the Pib gene-containing single-gene identification system shows moderate resistance to the 'Sichuan-43'. The five pairs of SSR pleomorphism markers comprise OJL03, OJL08, FJK08, P4F05 and FJK42, and are tightly interlocked with a target gene, so that the selecting efficiency and the veracity of the auxiliary selective breeding of rice blast molecular markers can be preferably improved.

Description

technical field [0001] The present invention relates to the rice blast resistance gene Pi-y43(t) of japonica rice Yunyin and its related 5 pairs of SSR polymorphic markers. Based on fine mapping and map-based cloning of Pi-y43(t), compared with the known resistance gene Pib, Yunyin containing Pi-y43(t) is highly resistant to "Sichuan-43" blast disease strain , while the single-gene differential line containing Pib gene showed moderate resistance to "Sichuan-43". Background of the invention [0002] Rice blast is caused by a fungus (Magnaporthe oryzae) and is one of the most serious diseases widely occurring in various rice regions in the world. The annual loss of rice due to rice blast accounts for about 10% to 15% of the total output, and the loss amounts to billions of dollars. The current ways to control the prevalence of rice blast are mainly to use pesticides for chemical control and to breed new rice varieties resistant to rice blast. Pesticides are very harmful to ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C07K14/415C12Q1/68C12N15/11
Inventor 张建福谢华安陈锦文何炜连玲朱永生
Owner 福建省农业科学院水稻研究所
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