Gene for regulating and controlling corn drought stress resistance and application thereof

A corn and genetic technology, applied in the fields of application, genetic engineering, plant genetic improvement, etc., can solve the problems of difficult production and utilization

Inactive Publication Date: 2017-03-08
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Recent studies have shown that controlling ABA signaling is one of the important mechanisms for maize to respond to drought stress, in which PYR / PYL / RCAR proteins, cluster A PP2C phosphatases (PP2CA) and type 3 SnRK2 kinases form ABA receptors or other protein complexes. Drugs mediate the early transmission of ABA signaling, but the natural variation of its related coding genes and their mechanism of drought resistance are still poorly understood
In addition, the identified drought-resistant genes in maize are mainly based on transgenic overexpression or deletion mutants, but it is still difficult to use them in production

Method used

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  • Gene for regulating and controlling corn drought stress resistance and application thereof
  • Gene for regulating and controlling corn drought stress resistance and application thereof
  • Gene for regulating and controlling corn drought stress resistance and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1 Overexpression of ZmPP2CA10 weakens drought tolerance in maize

[0047] The full-length CDS of ZmPP2CA10 gene primers were designed using the reference sequence of maize B73 as a template, and part of the vector sequence was added to both ends of the primers, and the fragment was cloned in vitro using the B73 leaf cDNA as a template, and connected to the overexpression vector by homologous recombination pZZ0153-3HA, and sequenced to verify that the correct clone was obtained. The overexpression vector plasmid was sent to China Seed Group Co., Ltd. (Wuhan) to obtain the transgenic plants of the gene. The applicant obtained two homozygous overexpression ZmPP2CA10 maize transgenic material lines 6 and 12 through herbicide screening.

[0048] Extract the RNA of transgenic material family 6 and family 12, and wild-type C01 material, perform reverse transcription, and detect whether the target gene is overexpressed at the transcriptional level by fluorescent quanti...

Embodiment 2

[0052] Example 2 Identification of ZmPP2CA10 Preference Alleles Associated with Drought Tolerance

[0053] In order to further confirm the details of the gene mutation of ZmPP2CA10, the applicant randomly selected 100 materials from different inbred lines of maize to resequence the ZmPP2CA10 gene. Using the genomic DNA of these 100 materials as a template, primers were designed and amplified based on the B73 reference genome sequence, and the ZmPP2CA10 promoter region and gene region (including exons and introns) were measured respectively.

[0054] Multiple sequence alignments were performed on the ZmPP2CA10 sequence of each material, and all the SNP / InDel sites and the survival rate data of the associated population (the data came from previous articles published by others) were used for candidate gene association analysis, and a site InDel- 338 were significantly correlated with drought ( figure 2 A).

[0055] There is an InDel-308 site near the InDel-338 site. Through l...

Embodiment 3

[0061] Example 3 Luciferase experiments verify that the ERSE element in the promoter region is necessary for the activation of ZmPP2CA10 by endoplasmic reticulum stress signals, but does not affect the activation of ZmPP2CA10 by the ABA pathway

[0062] Clone the 1000bp promoter fragment upstream of the ZmPP2CA10 gene ATG in the CIMBL55 material. The InDel-338 site of ZmPP2CA10 in this material lacks the ERSE element. This fragment is named A10pΔERSE. The nucleotide sequence of the missing part is shown in SEQ ID NO: 1. After mutation The nucleotide sequence of the promoter of ZmPP2CA10 is shown in SEQ ID NO:2. A point mutation was made to A10pΔERSE, and the InDel-338 site was mutated to contain an ERSE element, which was named A10pERSE. The two fragments were ligated by restriction enzymes and connected to the pGL-Basic-LUC vector to construct pGL-Basic-A10pΔERSE-LUC and pGL-Basic-A10pERSE-LUC. The two plasmids were transformed into maize protoplasts and cultured overnight, ...

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Abstract

The invention relates to the technical field of plant genetic engineering, in particular to a gene for regulating and controlling the corn drought stress resistance. The gene is a ZmPP2CA10 deletion mutation gene; the deletion segment is a continuous base sequence with a distance being 301bp away from the upstream of the ZmPP2CA10 initial codon ATG; the continuous basic group sequence is shown as SEQ ID NO:1; the nucleotide sequence of the mutated ZmPP2CA10 promoter is shown as SEQ ID NO:2. The deletion mutation gene can be used for corn drought resistance relevant molecular marker auxiliary breeding and corn drought resistance auxiliary identification; the operation is convenient; the accuracy rate is high.

Description

technical field [0001] The invention relates to the technical field of plant genetic engineering, in particular to a gene regulating corn drought stress resistance and its application. Background technique [0002] Corn is one of the most important food crops. In my country, the planting area of ​​corn has surpassed that of rice, becoming the largest crop. Maize (Zea may L.) is a cross-pollinated annual grass plant belonging to the same plant with different flowers. Taxonomically, it belongs to Gramineae (Graminease) Zea family (Maydeae) Zea genus (Zea L.), originated in Central America, and was introduced to China in the early 16th century. Corn is not only an important food crop, but also an important feed and industrial raw material. It has been greatly favored as an energy crop in recent years. Changes in the natural environment have seriously affected the increase in corn yields. Drought is one of the main adversity factors affecting plant growth and crop yields, whic...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/55C12Q1/68
CPCC12N9/16C12Q1/6895C12Q2600/13C12Q2600/156C12Y301/03016
Inventor 代明球向艳丽孙霄鹏
Owner HUAZHONG AGRI UNIV
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