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Application of OsDBP1 protein in regulating cold tolerance of rice

A technology of cold resistance and protein, applied in the biological field, can solve the problems of reduced yield and no harvest, and achieve the effect of important application and promotion value.

Active Publication Date: 2019-04-16
INST OF BOTANY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Susceptible to cold damage caused by low temperature weather, resulting in reduced yield, especially when the seedlings are cold, it may lead to crop failure

Method used

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  • Application of OsDBP1 protein in regulating cold tolerance of rice
  • Application of OsDBP1 protein in regulating cold tolerance of rice
  • Application of OsDBP1 protein in regulating cold tolerance of rice

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] 1. Obtaining and identification of OsDBP1 mutant material osdbp1

[0044] Rice osdbp1 (PFG_1E-01523, T-DNA insertion mutant) and its corresponding wild-type rice Dongjin were obtained from the Korean Mutant Bank RISD (http: / / cbi.khu.ac.kr / RISD_DB.html). The Latin name of rice Dongjin is Oryza.sativa L.spp.japonica cv.Dongjin. The rice obtained is in the form of seeds.

[0045] Hull the rice seeds, then soak them in 70% alcohol aqueous solution for 30s, then soak them in 2.5% NaClO aqueous solution for 10-15min, then wash them with sterile water 4-5 times (10min each time), and then inoculate them in 1 / 2MS medium placed on a flat plate in a light incubator (light intensity 10000 μmol / m 2 / s, the light time is 16h / d, and the temperature is 30℃) for three weeks. Take seedlings, extract genomic DNA, and use genomic DNA as a template to carry out PCR amplification with LP / RP primer pair and RB / RP primer pair, respectively. LP: 5'-TCTTCCCATTCACACCTTCC-3'; RP: 5'-TCGAGCACG...

Embodiment 2

[0059] Example 2. Inhibiting OsDBP1 gene expression through RNAi interference to improve rice seedling cold tolerance

[0060] Rice Zhonghua No. 11, referred to as Zhonghua 11, is represented by ZH11.

[0061] Plant RNAi expression vector pTCK303, referred to as vector pTCK303, is the plasmid pTCK303 prepared in Example 1 of the patent whose application number is 03160092.1 (authorized announcement date is June 20, 2007; authorized announcement number is CN1322131C).

[0062] 1. Construction of recombinant plasmids

[0063] 1. Using rice seedlings at the 3rd leaf stage of Zhonghua No. 11, extract total RNA and reverse transcribe to obtain cDNA.

[0064] 2. Using the cDNA obtained in step 1 as a template, PCR amplification is performed using a primer pair composed of OsDBP1-RNAiF and OsDBP1-RNAiR, and the PCR amplification product is recovered.

[0065] OsDBP1-RNAiF:5'-GG GGTACCACTAGT CGCCTTCGCTTAATAATTCTCCT-3';

[0066] OsDBP1-RNAiR:5'-CG GGATCCGAGCTC CCTGATCGGATAACTGGG...

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Abstract

The invention discloses application of an OsDBP1 protein in regulating cold tolerance of rice and a method for cultivating a transgenic plant. The method comprises the steps of inhibiting expression of an osdbp1 gene in a starting plant to obtain the transgenic plant having enhanced cold tolerance. The invention also provides a plant breeding method comprising the steps of reducing the content and / or the activity of the OsDBP1 protein in a target plant, thereby increasing the cold tolerance of the target plant. The methods have important application and promotion value for rice breeding for the purpose of cultivating the cold-tolerant rice.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to the application of OsDBP1 protein in regulating the cold tolerance of rice. Background technique [0002] Rice (Oryza sativa L.) originated in tropical or subtropical regions and is a temperature-loving crop. High-yielding cultivars promoted today have limited tolerance to low temperatures. Susceptible to cold damage caused by low temperature weather, resulting in reduced yield, especially when the seedlings are cold, it may lead to a failure of harvest. Therefore, improving the cold tolerance of rice has important economic and practical significance. [0003] At present, some progress has been made in the research on rice low temperature signaling pathway. The low temperature response pathways in rice mainly include ABA-dependent signaling pathway, DREB-CRT / DRE pathway and ROS-activated kinase cascade amplification pathway. Contents of the invention [0004] The pu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82C12N15/29C07K14/415A01H5/00A01H6/46
CPCC07K14/415C12N15/8273
Inventor 种康夏昌选巩燕山徐云远
Owner INST OF BOTANY CHINESE ACAD OF SCI
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