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Gene related to chlorophyll content of leaf and grain weight of wheat as well as Indel marker and application thereof

A technology of chlorophyll content and gene, applied in the fields of genetic engineering and molecular biology, can solve the problems of undiscovered wheat BAS1 gene research report, incompletely clear mechanism of action, rare and other problems

Inactive Publication Date: 2015-12-09
ANHUI AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Although there are many studies on the structure and function of peroxiredoxin in plants, its specific mechanism of action in plants is still not completely clear.
Research reports on Prx in wheat are relatively rare, and no relevant research reports on wheat BAS1 gene have been found so far

Method used

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  • Gene related to chlorophyll content of leaf and grain weight of wheat as well as Indel marker and application thereof
  • Gene related to chlorophyll content of leaf and grain weight of wheat as well as Indel marker and application thereof
  • Gene related to chlorophyll content of leaf and grain weight of wheat as well as Indel marker and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1 Obtaining wheat TaBAS1-2B gene

[0041] The gene TaBAS1-2B related to wheat leaf chlorophyll content and grain weight was obtained by electronic cloning combined with conventional PCR. Search the wheat EST database according to the CDS sequence (GenBank: AB000405.1) of wheat thioredoxin peroxidase (Thiol-specific antioxidant protein) published on NCBI, and select the EST sequences (CA484935.1, CK214438.1, CK217637) .1) Electronic splicing, using the splicing sequence as a template, using PrimerPremier5.0 software to design three pairs of primers B1, B2, and B3 for obtaining wheat BAS1 gene cDNA, and the other two pairs of primers G1 and G2 for obtaining wheat BAS1 gene , The primer information is shown in Table 1.

[0042] Table 1 Primers used to amplify the wheat TaBAS1-2B gene and its cDNA sequence

[0043]

[0044] The PCR reactions were carried out on the BIO-RADMyCycler1.0 PCR machine using the cDNA and genomic DNA of wheat variety ‘Jing411’ as templates. Aft...

Embodiment 2

[0071] *Example 2 Development of wheat TaBAS1-2B gene functional marker

[0072] According to the sequence of the gene TaBAS1-2B cloned in Example 1, a polymorphic Indel marker was developed between the two varieties of'Jing411' and'Hongmangchun 21', and a pair was designed using PrimerPremier5.0 software. To amplify the Indel-labeled primer and analyze its effect on the phenotype in 194 natural wheat varieties, and finally develop a co-segregation with the gene TaBAS1-2B, and closely match the chlorophyll content and grain weight of wheat leaves Related function mark.

[0073] The primer pair used to amplify the Indel label is:

[0074] Upstream primer F: 5’-CGCAGTGCCTGTCGTTTC-3’ (SeqIDNo.5)

[0075] Downstream primer R: 5’-TCACATACTTCTTCCCAAT-3’ (SeqIDNo.6)

[0076] The characteristic band of Indel marker that is related to the higher chlorophyll content and higher grain weight of wheat leaves amplified by the primer pair is 494 bp, and its nucleotide sequence is shown in SeqID No. ...

Embodiment 3

[0077] Example 3 Correlation between different genotypes of wheat TaBAS1-2B gene and flag leaf chlorophyll content and grain weight after anthesis

[0078] 1. Determination of chlorophyll content and thousand-grain weight of wheat at different stages after anthesis

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Abstract

The invention relates to a gene TaBAS1-2B related to the chlorophyll content of a leaf and the grain weight of wheat and an Indel marker of the gene. According to the gene, the gene TaBAS1-2B related to the chlorophyll content of the leaf and the grain weight of the wheat is separated and cloned from wheat for the first time, the Indel marker with polymorphism is developed between two varieties of Jing 411 and Hongmangchun 21 according to a cloned TaBAS1-2B gene sequence, the effect on phenotype is analyzed in 194 natural wheat varieties, and finally, a functional marker which is separated together with the gene TaBAS1-2B and is closely related to the chlorophyll content of the leaf and the grain weight of the wheat is developed. The Indel marker can be used for respectively explaining 8.2% and 4.4% of phenotypic variation of the chlorophyll content of a flag leaf and the thousand seed weight after wheat anthesis, wherein a strip of which the molecular weight is 494bp (Seq ID No.3) has a synergistic effect on the increase of the grain weight, and a strip of which the molecular weight is 493bp (Seq ID No.4) has a synergistic effect on the reduction of the grain weight. The development of the molecular marker provides a practical way for assistant breeding of high-yield wheat molecules.

Description

Technical field [0001] The present invention relates to the fields of genetic engineering and molecular biology, in particular to genes related to chlorophyll content and grain weight of wheat leaves, their Indel markers and applications. Background technique [0002] Wheat is one of the most important food crops in the world. With the increase of population, the reduction of arable land and the continuous increase of food production costs, high-yield and super-high-yield breeding has become an urgent problem in wheat breeding in my country. The growth process of wheat is a process of continuous material production through photosynthesis, and green leaves are the main source of material production. Under suitable conditions, about 70-90% of wheat grain yield during grain filling is derived from photosynthetic products produced by flag leaves after anthesis (Austin et al., 1997; Bidinger et al., 1997). Therefore, extending the functional period of wheat flag leaves and promoting ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/53C12N15/11C12Q1/68
Inventor 朱晓峰常成张海萍曹佳佳张衡张礼廉曹雪莲卢杰司红起马传喜
Owner ANHUI AGRICULTURAL UNIVERSITY
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