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A kind of rapeseed drought tolerance negative regulator gene and its application

A gene and drought resistance technology, applied in application, genetic engineering, plant genetic improvement, etc., can solve problems such as insufficient understanding of the function and role of NAC transcription factors

Active Publication Date: 2022-07-05
HENAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The NAC transcription factor family plays an important role in plant resistance to various abiotic stresses. However, the functions and roles of NAC transcription factors in response to abiotic stress in rapeseed are not well understood

Method used

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  • A kind of rapeseed drought tolerance negative regulator gene and its application
  • A kind of rapeseed drought tolerance negative regulator gene and its application
  • A kind of rapeseed drought tolerance negative regulator gene and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 Cloning of BnNAC038 Sequence of Drought Sensitive Protein Coding Gene of Brassica napus

[0032] The specific method for extracting total RNA from Brassica napus seedlings was extracted using RNA extraction and separation reagent (Trizol, Invitrogen). Put the frozen samples into a high-throughput tissue grinder and grind at 45Hz for 40s; after grinding, add 1 mL of Trizol solution to each EP tube, invert several times by hand, shake well, and place on ice for 5 min; add 200 μL of Trizol solution to each EP tube Chloroform was added to each sample, mixed slowly and then kept on ice for 5 min; the above samples were centrifuged at 4°C and 12,000 rpm for 10 min; the supernatant was sucked into a new RNase free EP tube with a pipette, Add an equal volume of -40°C pre-cooled isopropanol to the supernatant, invert up and down and mix evenly at 4°C, 12,000 rpm, and centrifuge for 15 min; discard the supernatant, first add 250 μL of absolute ethanol, and then add 750 ...

Embodiment 2

[0037] Example 2 Acquisition of transgenic BnNAC038 gene plants

[0038] 1. Construction of plant overexpression vector of Brassica napus BnNAC038 gene: The overexpression vector of the gene was constructed by Gateway recombination technology. First, the RFP gene was added to the pEarleyGate103 vector to transform it, and the Arabidopsis-specific promoter At2S3 was used to express it. , the modified vector was named pEarleyGate103-RFP. The target gene was linked to pDONOR 207 vector by BP reaction (BP Clonase II Enzyme Mix, Invitrogen) to form a recombinant plasmid containing the target gene. The correctly sequenced recombinant plasmid was subjected to LR reaction (LR Clonase II Enzyme Mix, Invitrogen No. 11791-020) with the pEarleyGate103-RFP expression vector, the BnNAC038 gene was recombined into the pEarleyGate103-RFP vector, and transformed into E. coli DH5α competent cells, The successfully recombined overexpression vector pEarleyGate103-RFP-BnNAC038 was screened by 50m...

Embodiment 3

[0046] Example 3 Detection of drought sensitivity in Arabidopsis thaliana transgenic BnNAC038

[0047] 1. Detection of drought tolerance in transgenic Arabidopsis overexpressing BnNAC038

[0048] The Arabidopsis thaliana seedlings grown in the soil for 3 weeks with normal watering were selected to study the tolerance of drought / osmotic stress. Most of the wilting and water loss were more serious than WT. After 3 days of rehydration, the drought and wilting of WT were relieved, and the leaves returned to green, but most of the transgenic plants could not recover, and the survival rate was significantly lower than that of WT. The water loss rate of the isolated leaves of the plants was measured, and the results showed that the leaves of the overexpressed BnNAC038 transgenic lines lost water faster than the wild type. The results are as follows: Figure 4 shown.

[0049] 2. Determination of relevant physiological indicators of overexpressed BnNAC038 transgenic Arabidopsis

[0...

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Abstract

The invention belongs to the technical field of plant molecular biology, and in particular relates to a negative regulation gene for drought tolerance of rape and its application. The gene is the BnNAC038 gene, and the nucleotide sequence is shown in SEQ ID NO. 1. The rape drought tolerance negative regulator gene BnNAC038 provided by the present invention plays a negative regulatory role in drought resistance, and its overexpression in the transgenic plant will inhibit the coding participation. The expression of genes in response to osmotic or oxidative damage induced by abiotic stress reduces the drought resistance of transgenic plants, providing a theoretical basis and related genes for the subsequent cultivation of drought-tolerant transgenic crops.

Description

technical field [0001] The invention belongs to the technical field of plant molecular biology, and in particular relates to a negative regulation gene for drought tolerance of rape and its application. Background technique [0002] Brassica napus, as an important oil crop in my country, is the main cultivar in rapeseed oil production. Brassica napus is an allodiploid, derived from the hybridization of Brassica napus and turnip, with oil value, feed value and ornamental value. In recent years, the planting area of ​​rapeseed has increased year by year, but the stress resistance of rapeseed is poor, and the frequent occurrence of extreme weather such as drought and low temperature has seriously affected the yield of rapeseed. Especially in the next few years, with the severe global warming problem, the water content in the atmosphere will gradually increase, the water content in the soil will decrease, and there will be a high frequency of drought stress. Therefore, through...

Claims

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Application Information

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IPC IPC(8): C12N15/29C12N15/82C07K14/415A01H5/00A01H6/20A01H6/82
CPCC07K14/415C12N15/8273
Inventor 王道杰陈玉平杨翠玲刘雷迪王勇锋席梦微丁群英
Owner HENAN UNIVERSITY
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