Magnaporthe grisea exciton CSB I gene and clone method thereof
A cloning method and rice blast fungus technology, applied in genetic engineering, plant genetic improvement, botanical equipment and methods, etc., can solve the problems of easy mutation, single variety of rice blast resistance, severe susceptibility to disease, etc.
Inactive Publication Date: 2012-01-25
SOUTH CHINA AGRI UNIV
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However, due to the complex genetic background of Magnaporthe oryzae, it is easy to mutate, and the blast-resistant varieties are relatively single, and the genetic homogeneity of the main cultivars makes it difficult for disease-resistant breeding to keep up with the variation speed of the physiological races of Magnaporthe oryzae, which often leads to A new blast-resistant rice variety became severely susceptible 3 to 5 years after being planted on a large scale
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[0031] The present invention will be further described in detail below in conjunction with specific embodiments and accompanying drawings.
[0032] According to the tested blast fungus ZC 13 Primers were designed for the coding sequence of CSB I elicitor peptide in physiological race, the full-length sequence of CSB I cDNA was cloned, and the amino acid sequence of CSB I was deduced according to the coding rules. Include the following steps:
[0033] (1) total RNA is extracted after the cultivation of the physiological race strain of Magnaporthe grisea;
[0034] (2) Using (1) total RNA as a template, the first strand of cDNA was synthesized by RT-PCR; the deduced cDNA sequence of the CSB I elicitor of M. oryzae physiological race was cloned by using the rapid amplification technology of cDNA ends.
[0035] Step (1) process is as follows:
[0036] Magnaporthe grisea ZC 13 Physiological race strains were activated and then inoculated in liquid culture for 4 days at 25°C. The...
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The invention discloses CSB I gene of rice blast fungus elicitor and a cloning method thereof. The nucleotide sequence encoded by the gene comprises the sequence of the total length of cDNA as SEQ ID NO: I and amino acid sequence as SEQ ID NO: 2. Primers are designed according to the coding sequence of peptide end of the CSB I elicitor which has been measured. Total RNA of physiological race of rice blast fungus ZC13 is taken as template, and first strand of cDNA can be synthesized by RT-PCR method. The deduced cDNA sequence of the CSB I elicitor of physiological race of the rice blast fungus ZC13 can be achieved by utilizing the fast amplified technology to clone. The invention can be the basement of the mechanism of wide and abundant researches on the producing disease assistant activity of the rice induced by the elicitor. Besides, the invention can also be used for research on transgenic breeding of the rice.
Description
technical field [0001] The invention relates to the field of biotechnology, in particular to a blast fungus glycoprotein elicitor CSB I gene and a cloning method thereof. Background technique [0002] Rice blast disease (rice blast disease) is one of the three major diseases of rice caused by the fungus Magnaporthe grisea (Hebert) Barr [its asexual generation is Pyricularia grisea (Cooke) Sacc.], widely distributed and serious in rice cultivation countries and regions affect rice yield and quality. Rice blast occurs to varying degrees all year round, and in popular years, the yield is generally reduced by 10%-20%, and when it is severe, it reaches more than 50%. Breeding and utilization of rice blast-resistant varieties is an effective measure to control rice blast. However, due to the complex genetic background of Magnaporthe oryzae, it is easy to mutate, and the blast-resistant varieties are relatively single, and the genetic homogeneity of the main cultivars makes it di...
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IPC IPC(8): C12N15/09C12N15/29
Inventor 王振中童英林董章勇纪春艳李云锋
Owner SOUTH CHINA AGRI UNIV