Molecular marker of rice resistance gene Xa23 and application of molecular marker

A molecular marker and resistance gene technology, applied in the biological field, can solve problems such as loss of resistance to bacterial blight resistance genes, pathogenic variation of pathogenic bacteria, etc., achieve high reliability, and eliminate the effect of field inoculation identification work.

Active Publication Date: 2015-07-22
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Long-term widespread utilization of a small number of bacterial blight resistance genes is always at risk of loss of resistance due to variation in pathogenicity

Method used

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  • Molecular marker of rice resistance gene Xa23 and application of molecular marker
  • Molecular marker of rice resistance gene Xa23 and application of molecular marker
  • Molecular marker of rice resistance gene Xa23 and application of molecular marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Example 1 Determination of Lj74 Molecular Marker

[0020] 1 plant material

[0021] The susceptible reincarnated parent King Kong 30 (JG30) is the female parent, carrying the bacterial blight resistance gene Xa23 The near isogenic line CBB23 was used as the male parent. Crossing JG30 with CBB23, F 1 Single harvest. f 1 F 2 A total of 2,562 individual plants were obtained from the isolated populations from generation to generation, and were planted in the net room of the Institute of Crop Science, Chinese Academy of Agricultural Sciences, under conventional water and fertilizer management.

[0022] Rice bacterial blight

[0023] Rice bacterial blight wide-pathogenic strain PXO99 (called P6) was introduced from the International Rice Research Institute (IRRI), stored in vacuum at –70°C, rejuvenated on Wakimoto Tetsu’s medium before use, and placed at 28°C Cultivate for 48 hours, prepare the inoculum solution with sterile water, and adjust the concentration t...

Embodiment 2

[0042] Example 2 Verification of molecular markers

[0043] In order to further verify whether the molecular marker Lj74 is only for the molecular marker of the Xa23 gene, molecular detection was carried out on varieties containing different genes and some varieties currently being promoted.

[0044] Since primer Lj74 is with Xa23 Gene co-segregation markers, only containing Xa23 The variety of the gene can expand the DNA fragment consistent with the CBB23 band pattern, which does not contain Xa23 The extended DNA band pattern of the variety was inconsistent with the CBB23 band pattern. Therefore, we selected 25 cultivars containing different bacterial blight resistance genes that have been mapped and cloned, including IR24 (no resistance gene), IRBB1 ( Xa1 ), IRBB2 ( Xa2 ), IRBB3 ( Xa3 ), IRBB4 ( Xa4 ), IRBB5 ( xa5 ), IRBB7 ( Xa7 ), IRBB8 ( xa8 ), IRBB10 ( Xa10 ), IRBB11 ( Xa11 ), IRBB13 ( xa13 ), IRBB14 ( Xa14 ), M41 ( xa15 ), Tetep ( Xa16 ),...

Embodiment 3

[0046] Example 3 Molecular markers Lj74 sequencing

[0047] In order to further study the molecular marker Lj74 is the sequence difference between the resistant variety and the non-resistant variety, the molecular marker Lj74 determined in the embodiment example, that is, using SEQ ID NO: 1 and 2 as the primer amplified fragment was sequenced. The fragment sequence amplified from the resistant variety CBB23 is shown in SEQ ID NO: 3, and the fragment sequence amplified from the resistant variety CBB23 is shown in SEQ ID NO: 4.

[0048] After comparing the files, it is found that the resistant variety has two copies of the nucleotide sequence fragment shown in SEQ ID NO: 6, that is, the nucleotide sequence fragment shown in SEQ ID NO: 5, while the non-resistant variety has only one copy A fragment of the nucleotide sequence shown in SEQ ID NO:6. Based on this finding, those skilled in the art can design primer pairs or probes based on the sequence for detection (not limited t...

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Abstract

The invention discloses a molecular marker of a rice bacterial leaf blight resistance gene Xa23. The molecular marker is not only closely linked to the rice bacterial leaf blight resistance gene Xa23, but also is co-separated from the resistance gene in plant offspring, so that a good way is provided for resistant rice variety screening and molecular marker-assisted breeding.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a rice bacterial blight resistance gene molecular marker. Background technique [0002] by Xanthomonas ( Xanthomonas oryzae PV .oryzae, Xoo ) caused by rice bacterial blight is one of the main diseases in rice production, seriously affecting the yield and quality of rice. Practice has proved that using the disease resistance of rice varieties is the most economical and effective way to control rice bacterial blight, and discovering and using new genes resistant to bacterial blight has always been one of the research hotspots. In the early days, it mainly started with cultivated varieties, and later excavated from wild resources [Brar DS and Khush GS.1997, , Alien introduction in rice. Plant Mol. Biol. 1997,35:35-47; Natraj KP,2012, Identification and fine -mapping of Xa33, a novel gene for resistance to Xanthomonas oryzae pv. oryzae. Phytopathology, 2012, 102:222-228]. So far, 36 ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11C12Q1/68A01H5/00
Inventor 赵开军王春连樊颖伦
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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