Application of oryza sativa l. OsGA2ox8 protein and coding gene and recombinant vector thereof in enhancing plant drought resistance

A technology for encoding gene and drought resistance, which is applied to rice OsGA2ox8 protein and its encoding gene and recombinant vector to enhance the application field of plant drought resistance

Inactive Publication Date: 2019-11-05
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

However, the research on the OsGA2ox gene mainly focuses on salt stress resistance, and it is not known how it will perform in drought resistance

Method used

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  • Application of oryza sativa l. OsGA2ox8 protein and coding gene and recombinant vector thereof in enhancing plant drought resistance
  • Application of oryza sativa l. OsGA2ox8 protein and coding gene and recombinant vector thereof in enhancing plant drought resistance
  • Application of oryza sativa l. OsGA2ox8 protein and coding gene and recombinant vector thereof in enhancing plant drought resistance

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Spatiotemporal expression differences of OsGA2ox8 gene under different stress conditions

[0030] Experimental material: Nipponbare (Oryza sativa ssp.japonica) (obtained from the Institute of Crop Science, Chinese Academy of Agricultural Sciences);

[0031] 1. Low temperature, high salinity and drought stress of rice

[0032] The rice Nipponbare seeds are sterilized, soaked at room temperature for 2 days, and germinated at 37°C for 1 day. After germination, the seeds with consistent germination are selected and sowed on the foam frame of a plastic box, 2 seeds per hole, hydroponic before the second leaf, and cultivated with Yoshida nutrient solution after the second leaf . Low temperature (4°C), high salt (150mmol / LNaCl) and osmotic stress (20% PEG) treatments were started at the five-leaf stage. Nipponbare took leaves and roots after 0h, 1h, 3h, 6h, 12h, and 24h of stress treatment.

[0033] 2. Real-time quantitative PCR analysis

[0034] Use TRIZOL reagent to extra...

Embodiment 2

[0050] Overexpression of OsGA2ox8 gene enhances drought resistance of transgenic rice

[0051] 1. Construction of OsGA2ox8 gene overexpression vector

[0052] First, pGWC vector (publicly available from Institute of Crop Science, Chinese Academy of Agricultural Sciences) was digested with restriction endonuclease EmaI1051, linearized and recovered.

[0053] Primers were designed according to the full-length Nipponbare sequence, and the full-length coding region of the OsGA2ox8 gene was amplified using Nipponbare cDNA as a template. At the same time, the linearized adapters of the pGWC vector were added to the 5' and 3' ends. The amplification primers were:

[0054] F: 5′- gcaggctttgacttt ATGGTGGCGATCACGGCGC-3' (the underlined part is the linker sequence);

[0055] R: 5′- gggtctagagacttt TTTCTTCGTCGCGGCCTCATC-3' (the underlined part is the linker sequence); PCR amplification and recovery of the target DNA fragment (1092bp).

[0056] The In-Fusion HD Cloning Kit (Clontech,...

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Abstract

The invention provides an application of an oryza sativa l. OsGA2ox8 protein and a coding gene and a recombinant vector thereof in enhancing plant drought resistance, and belongs to the technical field of biological gene engineering. The OsGA2ox8 protein has the amino acid sequence shown in SEQ ID NO.1. Under drought, high salt and low temperature stress, along with prolonging of the treatment time and the change of the expression quantity of an OsGA2ox8 gene, and with combination of phenotypic change of overexpressed and CRISPR-knockout oryza sativa l. plants, a result shows that the OsGA2ox8gene in leaves is induced to perform up-regulated expression by drought and cold stress, and is inhibited to perform down-regulated expression in roots by drought and cold stress. In addition, whether in the roots or in the leaves, OsGA2ox8 is induced to perform up-regulated expression by salt stress, but the up-regulated range in the roots is small; with overexpression of the OsGA2ox8 gene in oryza sativa l., the ability of the oryza sativa l. to resist drought stress can be significantly improved.

Description

technical field [0001] The invention belongs to the technical field of biogenetic engineering, and specifically relates to the application of rice OsGA2ox8 protein, its coding gene and recombinant vector in enhancing plant drought resistance. Background technique [0002] Rice (Oryza sativa L.) is a cereal crop with high economic and social value. Together with wheat and corn, it feeds more than half of the world's population. It is the main food crop in my country and is crucial to ensuring food security in my country. However, with the exploration of rice yield potential, there is a sharp contradiction between high input for high yield and sustainable development of resources and environment. In recent years, the increasingly serious salinization of soil or irrigation water has become a major obstacle to high rice yield and further expansion of planting area. [0003] Early studies have shown that the stress tolerance of plants is extremely sensitive to plant hormones, an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/02C12N15/53C12N15/82A01H5/00A01H6/46
CPCC12N9/0071C12N15/8273C12N2800/60C12Y114/11013
Inventor 傅彬英王银晓王文生赵秀琴杜丰平徐建龙黎志康
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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