Plant disease-resistant protein Xa23 and its coding gene and use

A technology of disease-resistant protein and coding gene, which is applied in the direction of plant gene improvement, application, plant peptide, etc., and can solve the problem of loss of bacterial blight resistance gene resistance and other problems

Active Publication Date: 2013-10-02
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

The long-term widespread utilization of a small number of bacterial blight resistance genes always fac

Method used

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  • Plant disease-resistant protein Xa23 and its coding gene and use
  • Plant disease-resistant protein Xa23 and its coding gene and use
  • Plant disease-resistant protein Xa23 and its coding gene and use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1 Separation of Xa23 Candidate gene sequence cloning

[0030] 1 Plant materials

[0031] contain Xa23 The near-isogenic line of the gene is CBB23.

[0032] Carrier

[0033] The vector used to construct the BAC library of CBB23 is Epicentre's CopyControl TM pCC1BAC TM , The transformable artificial expression vector pYLTAC747 (provided by Dr. Liu Yaoguang of South China Agricultural University), the RNAi interference expression vector pTCK303 (WANG Z et al. A Practical Vector for Efficient Knockdown of Gene Expression in Rice ( Oryza sativa L.). Plant Molecular Biology Reporter 22: 409-417).

[0034] 3 BAC library construction

[0035] Will carry Xa23 The seeds of the near-isogenic line CBB23 of the gene were cultured in a dark room at 28-30°C to obtain chlorophyll seedlings. After being ground into powder with liquid nitrogen, they were gently stirred in a pre-cooled extraction buffer and then sprayed on 50 mesh, 200 mesh, and 400 mesh nylon. Mesh screen s...

Embodiment 2

[0044] Example 2 Obtained by rice genetic transformation and expression analysis Xa23 gene

[0045] 1 Plant materials

[0046] The recipient susceptible rice variety Mudanjiang 8 (MDJ8), the recipient resistant rice variety Zhongwu 1 (containing Xa23 gene).

[0047] Rice bacterial blight

[0048] PXO99, the pathogenic bacterial blight of rice used for inoculation and identification, was quoted from the International Rice Research Institute (IRRI), stored in vacuum at -70°C, rejuvenated on Tetsuji's medium Wakimoto before use, and placed at 28°C Cultivate for 48h, prepare inoculation bacterial solution with sterile water, and adjust the concentration to OD=1.0.

[0049] Agrobacterium-mediated genetic transformation

[0050] Combine the above T189 and other 10 containing Xa23 The TAC clone plasmid DNA of the candidate genes was transformed into Agrobacterium EH105 competent cells, and the 10 clones were transformed into the recipient susceptible variety Mudanjiang No. 8 by Agrobact...

Embodiment 3

[0070] Example 3 Transient expression proves that Xa23 protein causes allergic reactions in dicot cells

[0071] 1 Plant materials

[0072] Benming Nicotiana benthamiana ) Planted in an artificial climate room, culture conditions: light 16h / d, light intensity 30~40μmols -1 m -2 , The temperature is 25~20℃, and the humidity is 50~60%; the seeds of the original tobacco are directly sown in the nutrient soil. After two cotyledons grow, the seedlings are transferred and moved to the new nutrient soil (add equal volume of vermiculite, Conducive to soil ventilation), move 3 to 4 plants per pot, during the period, pay attention to prevent soil pests, watering and nutrient solution intervals.

[0073] Carrier

[0074] Fast PCR Clone Kit (Beijing Bioche-Service Sci and Tech Company), plant expression vector pCAMBIA1305.

[0075] Xa23 Construction of Gene Plant Expression Vector

[0076] The 35S promoter driver was constructed using the recombinant method (Fast PCR Clone) Xa23 Plant expres...

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Abstract

The invention discloses a paddy rice bacterial blight-resistant protein Xa23 and its coding gene. A nucleotide sequence of the coding gene is shown in the formula of SEQ ID NO: 1 and an amino acid sequence of the paddy rice bacterial blight-resistant protein Xa23 is shown in the formula of SEQ ID NO: 2. The invention also relates to a use of the coding gene in cultivation of disease-resistant crops and especially in cultivation of a paddy rice bacterial blight-resistant paddy rice variety.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a rice disease-resistant protein and its coding gene, in particular to a rice bacterial blight-resistant gene and its application. Background technique [0002] Rice is the most important food crop in my country. About 60% of the population takes rice as a staple food. The sustainable development of rice production is directly related to my country's food security. Rice bacterial blight (Bacterial blight, BB) is one of the major diseases in rice production in the world. It was first discovered in Fukuoka, Japan in 1884. Since the 1950s, the incidence range has continued to expand. The current occurrence range of bacterial blight All over the rice producing areas in the world (Mew TW. Current status and future prospects of research on bacterial blight of rice. Annu Rev Phytopathol, 1987, 25:359-382; Ou SH. Rice disease, Commonwealth Agricultural Bereaux, England, 1985, 380pp.). Rice b...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/82C12N15/84A01H5/00
Inventor 赵开军王春连张晓平樊颖伦高英秦腾飞
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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