Molecular marker method for anti-rice blast gene Pi-hk1 (t) of paddy

A rice blast resistance gene, pi-hk1 technology, applied in the field of molecular genetics, can solve the problems of difficult disease resistance gene aggregation, difficult phenotype identification, low breeding efficiency, etc., and achieves convenient identification, control of breeding population size, and high cost. Effect

Inactive Publication Date: 2011-08-17
NANJING AGRICULTURAL UNIVERSITY
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Problems solved by technology

However, traditional breeding methods are time-consuming, laborious, phenotypic identification is difficult, and breeding efficiency is low. Since most dis

Method used

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  • Molecular marker method for anti-rice blast gene Pi-hk1 (t) of paddy
  • Molecular marker method for anti-rice blast gene Pi-hk1 (t) of paddy
  • Molecular marker method for anti-rice blast gene Pi-hk1 (t) of paddy

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Embodiment 1

[0026] (1) Materials and methods:

[0027] 1. Li Peifu et al. obtained F 1 , selfing to obtain F 2 The population was isolated for mapping, and the Pi-hk1(t) gene was mapped between SSR markers RM7654 and RM27381 (Chinese Rice Science, 2007, 21: 579-584). On this basis, the present invention selects 3 homozygous disease-resistant single plants containing target DNA fragments for planting, respectively backcrossing and selfing with Su Yunuo to form BC 2 f 2 Separation of large groups for fine-tuning.

[0028] 2. For the methods of strain cultivation and inoculation identification, refer to Li Peifu et al. (Chinese Rice Science, 2007, 21: 579-584).

[0029] 3. DNA extraction method: use the SDS method to extract the DNA of each individual plant in the isolated population.

[0030] 4. Development of new markers: New markers were designed between the initial target area RM7654-RM27381.

[0031] (1) SSR marker design: download the genome sequence of the BAC where the target g...

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Abstract

The invention belongs to the field of molecular genetics, and discloses a molecular marker method for anti-rice blast gene Pi-hk1 (t) of paddy. The method comprises the following steps of: (1) selecting a paddy sample, and extracting genome deoxyribonucleic acid (DNA) of the paddy sample; and (2) performing polymerase chain reaction (PCR) amplification on the genome DNA of the paddy sample by utilizing any one, two or three pairs of molecular marker primers in s01, in5 and in3, performing electrophoretic detection on a PCR amplification product, and if corresponding DNA fragments of molecularmarkers are amplified, indicating that the Pi-hk1 (t) gene exists. In the method, the selection efficiency of a single marker is over 99.92 percent, and the selection efficiency of any double markersis 100 percent. The resistance level of the gene on rice blast can be forecasted by detecting whether the paddy contains the gene or not by the molecular marker primers provided by the invention, so the selection efficiency of the rice blast resistance of the paddy is improved greatly, and the process for breeding for disease resistance is accelerated.

Description

technical field [0001] The invention belongs to the field of molecular genetics, and relates to a molecular marker method of the rice blast resistance gene Pi-hk1(t), in particular to a molecular marker method of the rice blast resistance gene Pi-hk1(t) of A. Molecular marker-assisted selective breeding of rice blast varieties and utilization of germplasm resources. Background technique [0002] Rice is an important food crop in my country. Rice blast caused by blast fungus is one of the most threatening diseases to rice production, and it occurs in rice production areas. It is estimated that the global annual damage caused by rice blast is about billions of dollars in losses (Jeon et al, 2003). my country loses hundreds of millions of kilograms of rice every year due to rice blast damage. In the popular years of rice blast, the yield loss caused is generally 10-20%, and can reach more than 50% in severe cases. Some fields even fail to harvest, and cause the quality of ri...

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 王建飞张红生鲍永美谢留杰张晓军黄骥王州飞吴云雨
Owner NANJING AGRICULTURAL UNIVERSITY
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