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Application of osa-miR529b and precursor gene thereof in improving bacterial leaf-blight resistance of rice

A gene and precursor technology, applied in the biological field, can solve the problems of easy loss of variety resistance, complex diversity, and fast mutation speed.

Active Publication Date: 2019-12-13
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 fast mutation rate and complex diversity of Xanthomonas rice pathogenic varieties, production practice shows that the large-scale promotion and planting of varieties carrying a single disease resistance gene will lead to the increase of dominant virulent races or the mutation of new virulent races. Very easy to lose resistance

Method used

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  • Application of osa-miR529b and precursor gene thereof in improving bacterial leaf-blight resistance of rice
  • Application of osa-miR529b and precursor gene thereof in improving bacterial leaf-blight resistance of rice
  • Application of osa-miR529b and precursor gene thereof in improving bacterial leaf-blight resistance of rice

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1, the acquisition of osa-miR529b

[0027] Put rice seeds (introduced disease-resistant line FF329, recurrent parent HHZ) in a seed soaking bag, accelerate germination at 37°C for 2 days, then transfer them to seedling trays, and transplant them in the field after 2 weeks. Plant 10 rows of each material, 8 plants per row . At the peak tillering stage, the plants were infected with Xanthomonas oryzae PXO99, and inoculated by the leaf-cutting method. Each plant was inoculated with 5-6 slices, and samples were taken at 0h, 12h, 24h and 36h after inoculation, and 2 samples were taken at each time point. repeated for a total of 16 samples. When collecting samples, the inoculated part of rice and the part extending down about 1 cm should be taken. After sampling, immediately freeze in liquid nitrogen and store at -80°C.

[0028] The plant total RNA extraction kit (DP432) from TIANGEN Company was used to extract the above 16 samples, and the extracted RNA was subject...

Embodiment 2

[0044] Embodiment 2, the acquisition and identification of transgenic plants

[0045] 1. Construction of osa-miR529b precursor gene overexpression vector

[0046] In the pCUbi1390 vector, " TGCAGGTCGAC " was replaced by the double-stranded DNA molecule shown in sequence 2 of the sequence listing to obtain the recombinant plasmid pUbi::premiR529b. The pCUbi1390 vector has been sequenced, as shown in sequence 3 of the sequence listing. The recombinant plasmid pUbi::premiR529b has been sequenced and verified, As shown in sequence 4 of the sequence listing.

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Abstract

The invention discloses an application of osa-miR529b from rice and a precursor gene of the osa-miR529b in improving the bacterial leaf-blight resistance of the rice. The invention relates to miRNA (osa-miR529b) shown in a sequence 1 in a sequence table. The invention also discloses the precursor gene of the miRNA. The invention also discloses a method for preparing a transgenic plant, wherein themethod comprises the following steps: overexpressing the miRNA in a plant to obtain a transgenic plant with improved disease resistance. The invention also discloses the method for preparing the transgenic plant, wherein the method comprises the following step of introducing the precursor gene into a plant to obtain the transgenic plant with improved disease resistance. The method can be used forimproving the bacterial leaf-blight resistance of the rice, has important significance for cultivating new bacterial leaf-blight resistant rice varieties, and is suitable for promotion and application.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to the application of osa-miR529b obtained from rice and its precursor gene in improving rice bacterial blight resistance. Background technique [0002] Rice is an important food crop in the world. Rice bacterial blight caused by Xanthomonas oryzae pv. oryzae is the main bacterial disease in rice production, especially in Asia. In the rice areas of South China, rice bacterial blight has a wide incidence area, high incidence and rapid infection, which seriously restricts the production safety of my country's grain. Breeding and planting disease-resistant varieties is an economical and effective measure to control rice bacterial blight. So far, 44 rice bacterial blight resistance genes have been officially reported, of which only genes Xa3, Xa4, Xa21 and Xa23 have been widely used in breeding. Due to the fast mutation rate and complex diversity of Xanthomonas rice pathogenic...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12N15/84A01H5/00A01H6/46
CPCC12N15/113C12N15/8281C12N15/8205C12N2310/14
Inventor 周永力卢家玲黎志康
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI