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Application of OsDUF6 gene to improvement of drought resistance of rice

A technology of drought tolerance and rice, which is applied in the field of genetic engineering, can solve the problems that family genes have not yet been studied, and achieve the effects of increased survival rate, increased ABA content change, and weakened membrane lipid peroxidation

Pending Publication Date: 2020-07-10
福建省农业科学院水稻研究所
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The functions of only a few genes in the rice DUFs family have been revealed, and most of the family genes have not been studied

Method used

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  • Application of OsDUF6 gene to improvement of drought resistance of rice
  • Application of OsDUF6 gene to improvement of drought resistance of rice
  • Application of OsDUF6 gene to improvement of drought resistance of rice

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Cloning analysis of embodiment 1 OsDUF6 gene

[0039] Using indica rice mutant material 75-1-127 and indica rice variety 93-11 as parents, reciprocal crossing was carried out to obtain F 1 Plants continue to self-cross to obtain F 2 group. Use F 2 The population was used as the experimental material, and the Nipponbare database was used as the reference sequence to analyze the candidate genes for the target location intervals of the indel markers InDel-106 and InDel-115, and the candidate gene LOC_Os06g44810( http: / / www.ricedata.cn / gene / ).

[0040] According to the Nipponbare reference genome, the sequence comparison analysis of the candidate genes was carried out, and the corresponding specific primers were designed. The specific primer sequences are as follows:

[0041]810F: 5'-ATGGAGTCACCCGCCGCGGGCG-3'; SEQ ID NO.1;

[0042] 810R: 5'-TCATTTTGCCTTCCTTGGCCTC-3'; SEQ ID NO.2.

[0043] high fidelity enzyme Max Super-Fidelity DNA Polymerase was used to amplify it....

Embodiment 2

[0055] Construction of embodiment 2 OsDUF6 gene overexpression vector

[0056] Ubiquitin-driven pCUbi1390 (vector map see Figure 4 ) as an overexpression vector, digested with Pst I, recovered, and obtained a linearized cloning vector fragment; select the restriction site (Pst I), and design corresponding primers (Ubi:810 over F, Ubi:810 over R) for OsDUF6 The gene is amplified, digested, and recovered, referring to Clon The MultiS One StepCloning Kit ligation kit is connected to the corresponding overexpression vector by one-step method. The vector ligation system is as follows: 5xCEMultiS Buffer 4 μL, linearized cloning vector fragment 6 μL, insert fragment amplification product 3 μL, MultiS 2 μL, ddH 2 O 5μL; mix well, and react at 37°C for 30min. Transformation, sequencing, obtaining overexpression vector plasmid (OsDUF6 gene overexpression vector construction strategy schematic diagram see Figure 5 ).

[0057] The specific primer sequences are as follows:

[005...

Embodiment 3

[0060] Embodiment 3 Agrobacterium transformation (freezing-thawing method)

[0061] Transform the constructed overexpression vector plasmid into the Agrobacterium competent cell EHA105 for subsequent genetic transformation tissue infection experiments. The specific steps are as follows:

[0062] (1) Preparation of Agrobacterium Competent EHA105

[0063] 1) Thaw Glycerolbacterium EHA105 stored at -80°C on ice, then streak in solid medium containing rifampicin (Rif) resistance (50 μg / mL) on an ultra-clean bench, and place at 28°C Dark culture 2-3d;

[0064] 2) Pick a monoclonal colony and add it to 700 μL of LB liquid medium containing Rif resistance (50 μg / mL) for shaking culture, 150 rpm, 28 °C for dark culture, after the bacteria are cultivated to a certain concentration, place it in a 50 mL Erlenmeyer flask to continue expand training;

[0065] 3) When the OD value of the cultured bacteria is within the range of about 0.4-0.6, treat the bacterial solution in an ice bath for...

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Abstract

The invention discloses an application of an OsDUF6 gene to improvement of drought resistance of rice, and belongs to the technical field of genetic engineering. Through cloning, an unknown functionalstructural domain gene OsDUF6 of rice is obtained, and through heredity transformation, an overexpression plant of the gene is obtained. The overexpression plant of the OsDUF6 is subjected to droughttreatment, and the inventor finds that compared with a wild type plant, the overexpression plant has the advantages that the survival rate is increased, the activity of antioxidase is improved, the membrane lipid peroxidation degree is reduced, the ABA content change is increased, and the content of active oxygen (ROS) is reduced, so that results indicate that overexpression of the OsDUF6 can notably improve the drought resistance of the rice.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, and more specifically relates to the application of OsDUF6 gene in improving drought tolerance of rice. Background technique [0002] Rice is one of the most important food crops in the world, feeds more than 60% of the world's population, and occupies a very important position in the food crops of our country and the world. However, rice yield loss is increasing day by day, and Ray et al. found that about one-third of rice yield loss was caused by climate change. Due to the variable global climate, the yield of rice is greatly affected by abiotic stresses such as drought, high temperature and heavy metals, among which drought has the most serious impact on rice production. In recent years, with the intensification of urban and rural water use and large-scale pollution of fresh water resources, and due to the uneven distribution of precipitation in time and space, rice water use has ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12N15/82C07K14/415A01H5/00A01H6/46
CPCC07K14/415C12N15/8273
Inventor 张建福姜身飞谢云杰何炜肖开转蔡秋华谢华安
Owner 福建省农业科学院水稻研究所
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