Rice bacterial blight resistance-related gene osduf6 and its application

A technology for bacterial blight resistance and rice bacterial blight, which can be applied in application, genetic engineering, plant genetic improvement and other directions, and can solve problems such as loss of variety resistance

Active Publication Date: 2022-08-02
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

Due to the high degree of variability of rice bacterial blight, after large-scale promotion and planting of disease-resistant varieties carrying a single major gene, potential toxic races will become dominant races or new toxic races will appear due to mutations, which can easily lead to Variety resistance loss

Method used

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  • Rice bacterial blight resistance-related gene osduf6 and its application
  • Rice bacterial blight resistance-related gene osduf6 and its application
  • Rice bacterial blight resistance-related gene osduf6 and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0095] Example 1. Cloning of rice OsDuf6 gene

[0096] 1. Acquisition of rice OsDuf6 gene sequence

[0097] The genomic DNA of the leaves of rice cultivar FF329 (high resistance to bacterial blight strain PXO99A) was extracted, and this DNA was used as a template, using forward primer OsDuf6-F: 5'-AGTGATAAGATTATAGTCTAAT-3' and reverse primer OsDuf6-R :5'-TTTATAGCAAAAAGAAATGTCGA-3', PCR amplification was performed with PrimeSTAR G×L DNA Polymerase (Code: R050A, Takara) to obtain the amplified product (that is, the OsDuf6 gene sequence, and its nucleotide sequence was SEQ ID No. .2).

[0098] 2. Acquisition of the coding sequence of the rice OsDuf6 gene

[0099] The total RNA of the leaves of rice variety FF329 was extracted, and the RNA was used as a template to synthesize cDNA using FastKing gDNADispelling RT SuperMix (Code: KR118, TIANGEN). -3' and OsDuf6-CDS-R: 5'-TTAGTGTCGTCGAGATAAGAGTG-3', PCR amplification was carried out with PrimeSTAR G×L DNA Polymerase (Code: R050A,...

Embodiment 2

[0100] Example 2. Construction of OsDuf6 Gene Complementary Vector and Gene Knockout Vector

[0101] 1. Construction of OsDuf6 gene complementary vector

[0102] The OsDuf6 gene complementary vector pCAMBIA1300-OsDuf6 was constructed comprising the OsDuf6 gene self-promoter-driven genome full-length sequence (SEQ ID No. 4, including 2221 bp promoter sequence, 2183 bp OsDuf6 gene sequence and 810 bp termination region sequence). The operation steps are as follows:

[0103] (1) Extracting the genomic DNA of the leaves of rice variety FF329, using the DNA as a template, using the forward primer OsDuf6-CP-F: 5'-TGAGCTGATCAAACAACGAAAG-3' and the reverse primer OsDuf6-CP-R: 5'-ATGCACAGGAATGTCGGTACTT -3', perform PCR amplification with PrimeSTAR G×L DNA Polymerase (Code: R050A, Takara) to obtain the amplified product (that is, the full-length sequence of the OsDuf6 gene self-promoter-driven genome. After sequencing, its nucleotide sequence is SEQ ID No. 4), and cut rubber for recyc...

Embodiment 3

[0124] Example 3, the acquisition of transgenic rice

[0125] 1. Acquisition of OsDuf6 gene complementary transgenic plants

[0126] The rice variety Huang Huazhan (HHZ), which is highly susceptible to bacterial blight strain PXO99A, was used as the recipient plant for preparing transgenic rice, and the pCAMBIA1300-OsDuf6 in Example 2 was used to prepare transgenic rice, and the blank vector pCAMBIA1300 was used as a control. Specific steps are as follows:

[0127] 1. Take out the mature seeds of Huanghuazhan (HHZ), remove the husks, and pick out the seeds with full, smooth and sterile spots for disinfection.

[0128] 2. Inoculate the sterilized seeds on the induction medium, cultivate in the dark for about 14 days at 28°C, and select callus with good appearance and good growth ability.

[0129] 3. The recombinant vector pCAMBIA1300-OsDuf6 constructed in Example 2 was introduced into Agrobacterium tumefaciens EHA105 to obtain recombinant bacteria.

[0130] 4. Take the recom...

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Abstract

The invention discloses a rice bacterial blight resistance-related gene OsDuf6 and its application. The present invention first discloses that the present invention first discloses that the protein with the amino acid sequence of SEQ ID No. 1 or the fusion protein obtained by linking a tag at the N-terminus or / and the C-terminus of the amino acid sequence shown in SEQ ID No. 1 is used in the regulation of plant protein Application in leaf blight resistance. The invention further discloses a method for cultivating transgenic plants with enhanced bacterial blight resistance. The invention finds that the rice OsDuf6 gene has the function of positively regulating the bacterial blight resistance of rice, can be used to improve the bacterial blight resistance of rice, and is of great significance for cultivating new bacterial blight resistant rice varieties.

Description

technical field [0001] The invention belongs to the technical field of plant genetic engineering, relates to the function and application of a rice disease resistance-related gene, and in particular relates to a rice bacterial blight resistance-related gene OsDuf6 and its application. Background technique [0002] Bacterial blight of rice is an important bacterial disease caused by Xanthomonas oryzae pv.oryzae, Xoo, which spreads all over the world. Generally, it can lead to a reduction of about 10% in rice production, and in severe cases, it can reduce production by 50%-60%. Breeding disease-resistant varieties with resistance genes is currently the most economical and effective measure to control rice bacterial blight. So far, 45 rice bacterial blight resistance genes have been reported at home and abroad ( http: / / www.shigen.nig.ac.jp / rice / oryzabase / gene / list ). However, the disease resistance genes from wild rice are difficult to use; some resistance genes only have...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C12N15/84C07K14/415A01H5/00A01H6/46
CPCC07K14/415C12N15/8281C12N15/8213
Inventor 周永力卢家玲曾丹黎志康
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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