Method for cloning piricula oryzae gene of rice

Abstract

The invention discloses a method for cloning a piricula oryzae gene. The method comprises the following steps of: 1) designing a primer according to a known piricula oryzae gene so as to obtain a primer pair; 2) performing polymerase chain reaction (PCR) amplification on the primer pair obtained in the step 1) by taking pyricularia grisea high-resistant paddy as a template so as to obtain PCR amplification products; 3) sequencing the PCR amplification products obtained in the step 2) so as to obtain the piricula oryzae gene; and 4) transferring the piricula oryzae gene into rice blast susceptible paddy, measuring a resistance spectrum of rice blast to obtain the piricula oryzae gene with a new resistance spectrum, namely the target piricula oryzae gene. Experiments of the method prove that: by the method, the piricula oryzae gene in wild paddy is directly separated and paddy varieties are transformed so as to obtain piricula oryzae paddy; and the resistance spectrum of transgenic plants is determined, and the piricula oryzae gene with a new resistance spectrum is screened out, so that the overall improvement on the paddy varieties is facilitated.

Description

technical field [0001] The invention relates to the field of plant genetic engineering, in particular to a method for cloning a rice blast resistance gene. Background technique [0002] Rice is one of the most important food crops in the world, and more than half of the world's population uses rice as the main food. Rice blast is one of the most important diseases of rice. The rice blast caused by rice blast reduces the output of rice every year by 11% to 30%, and when it is serious, it reduces the output by 40% to 50%, or even fails to harvest. [0003] Practice has proved that the use of disease-resistant rice varieties is one of the effective ways to deal with the damage of rice blast. So far, nearly one hundred blast resistance genes have been mapped in rice, and 14 genes have been cloned, which are Pib, Pita, Pi9, Pid2, Pizt, Pi2, Pi36, Pi37, Pi-km, Pi5, Pid3, Pb1, Pit, Pi-k h , except for the gene Pid2 which encodes a serine-threonine receptor-like kinase, the other...

AI Technical Summary

Problems solved by technology

[0004] At present, the cloning of the rice blast resistance gene mainly relies on map-based cloning. Although both the indica rice 9311 and the japonica rice Nipponbare have complete genome sequences, which brings great convenience to the map-based cloning of rice genes, the cloning of the rice blast resistance gene is still difficult. said, there are still many difficulties

For example, the identification of rice blast resistance for each individual plant in a fine-mapping population is not only a huge workload, but also the incidence of rice blast in the field is easily affected by the environment, resulting in statistical errors in traits and accurate positioning; in addition, from the sequenced According to the 9311 and Nipponbare genome sequences, most of the NBS-LRR genes exist in clusters. Even if the target gene can be located in a certain segment, it is difficult to judge which one of the clustered genes is the real disease resistance gene; Furthermore, the currently used rice varieties have a narrow genetic background, and there are not many blast resistance genes available, while wild rice contains abundant blast resistance genes. However, the research on wild rice is still in its infancy. , without a fine genetic map and physical map, it is obviously more difficult to clone the rice blast resistance gene using the map

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