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Application of rice transcription factor OsWRKY53 in negative regulation and control of cold tolerance of rice in booting stage

A technology of rice transcription factor and transcription factor, which is applied in application, genetic engineering, plant gene improvement, etc., can solve problems such as poor differentiation of branches and spikelets, decreased seed setting rate, and reduced number of grains per panicle, so as to improve rice booting. The effect of long-term cold resistance

Active Publication Date: 2021-04-02
NORTHEAST INST OF GEOGRAPHY & AGRIECOLOGY C A S
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] If rice suffers from low temperature and chilling damage during the booting stage, the differentiation of branches and spikelets will be poor, the number of grains per panicle will decrease, and the seed setting rate will drop sharply, which will easily cause rice yield reduction

Method used

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  • Application of rice transcription factor OsWRKY53 in negative regulation and control of cold tolerance of rice in booting stage
  • Application of rice transcription factor OsWRKY53 in negative regulation and control of cold tolerance of rice in booting stage
  • Application of rice transcription factor OsWRKY53 in negative regulation and control of cold tolerance of rice in booting stage

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1: Biological function verification of rice booting stage negative regulator OsWRKY53

[0039] Observation of wild rice variety Longjing 11, oswrky53 mutant plants and OsWRKY53 overexpression transgenic plants under the background of Longjing 11 found that the seed setting rate of OsWRKY53 overexpressing plants was significantly lower than that of the control material Longjing 11 (such as figure 1 , 2 shown). Compared with the control material Longjing 11, the seed setting rate of the oswrky53 mutant plants was basically the same (such as image 3 , 4 shown). This suggests that OsWRKY53 is likely to be involved in the regulation of cold tolerance at the booting stage of rice.

[0040] Using wild rice varieties Longjing 11, oswrky53 mutant plants and overexpression transgenic plants under the background of Longjing 11 as experimental materials, the distance between the flag leaf and the second leaf of the rice was -5 ~ 0cm ( That is, when the anthers are in ...

Embodiment 2

[0043] Example 2: Detection of GA content in rice anthers negatively regulated by OsWRKY53

[0044] (1) Determination of GA content in anthers of Longjing 11 and oswrky53 mutant plants under normal and low temperature treatment

[0045] The wild rice variety Longjing 11 and the oswrky53 mutant plants under its genetic background were used as experimental materials. The distance between the flag leaf and the second leaf of the rice was -8 ~ -6cm (that is, the anthers were in the meiosis state). During the division stage), the rice was treated at 18°C ​​for 15 days (the anthers were in the three-nucleated stage), and the anthers of about 30 rice main tillers were sampled, and the GA content in the anthers was detected by Thermo Scientific Ultimate 3000 UHPLC and TSQ Quantiva in Wuhan Lvjian Company .

[0046] The result is as Figure 10 As shown, □ means LJ11-NC, ■ means oswrky53-NC, Indicates LJ11-LT, Indicates oswrky53-LT. Low temperature stress will lead to the decrease ...

Embodiment 3

[0063] Example 3: Verification that OsWRKY53 directly binds to and inhibits the transcriptional activity of GA-related biosynthetic genes:

[0064] (1) Sequence analysis of GA-related biosynthetic genes

[0065] According to the 2000bp sequence of the OsGA20ox1, OsGA20ox3 and OsGA3ox1 promoter regions published in the Rice Genome Annotation Project, it was found that there were 2 W-boxes (TTGACC) in the 2000bp promoter region of the OsGA20ox1 gene and 1 W-box in the 2000bp promoter region of the OsGA20ox3 gene (TTGACC), there is a W-box (TTGACC) in the 2000bp promoter region of OsGA3ox1 gene such as Figure 12 shown.

[0066] (2) EMSA experiments verified that OsWRKY53 directly binds to GA-related biosynthetic genes

[0067] 1. Purify the OsWRKY53 protein with the Mbp tag, transform the plasmid into Escherichia coli BL21, pick a single clone and put it in 3ml of liquid LB medium containing kana antibiotics, shake overnight at 37°C. Put the activated bacterial solution into ...

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Abstract

The invention discloses an application of a rice transcription factor OsWRKY53 in negative regulation and control of cold tolerance of rice in a booting stage, aims to provide the new application of the rice transcription factor OsWRKY53 in negative regulation and control of the cold tolerance of the rice in the booting stage, and provides an important theoretical basis for increasing the yield ofthe rice under a low-temperature stress. According to the invention, a new function that the OsWRKY53 participates in regulating and controlling the cold tolerance of the rice in the booting stage isdiscovered, and a molecular mechanism that the OsWRKY53 negatively regulates and controls the cold tolerance of the rice in the booting stage is analyzed. The low-temperature treatment can cause thereduction of the GA content in the rice anther, but the GA content in an oswry53 mutant is obviously higher than that in a wild type. Experiments show that the OsWRKY53 can be directly combined with and inhibit the transcriptional activity of GA related biosynthetic genes OsGA20ox1, OsGA20ox3 and OsGA3ox1, so that the GA content in the rice anther is negatively regulated and controlled, and the cold resistance of the rice in the booting stage is negatively regulated and controlled.

Description

technical field [0001] The invention relates to a new application of rice transcription factor OsWRKY53. Background technique [0002] Rice is an important food crop, which is the staple food for more than half of the world's population. In recent years, due to the abnormal climate, severe weather such as low temperature often occurs in the supposedly hot summer, and chilling injury at the booting stage has become a major natural disaster. Therefore, solving the problem of chilling damage at the booting stage, mining cold-tolerant genes and analyzing their functions have very important practical significance for improving rice yield and ensuring food security. GA, namely gibberellin, is a common plant hormone, which is involved in the regulation of cold tolerance at the booting stage of rice. Low temperature treatment will lead to a decrease in GA content in rice anthers, poor pollen fertility, and a decrease in seed setting rate. The degree of restoration of the phenotype...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29A01H5/00A01H6/46
CPCC07K14/415C12N15/8273
Inventor 田晓杰卜庆云王臻昱李秀峰
Owner NORTHEAST INST OF GEOGRAPHY & AGRIECOLOGY C A S
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