Gene MS33 related to maize male nuclear sterility and application thereof in cross breeding

A technology of male sterility and male fertility, applied in application, genetic engineering, plant genetic improvement, etc., can solve problems such as limited nuclear sterility genes, and achieve the effect of complete traits

Active Publication Date: 2016-10-26
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, currently there are very limited genetic sterile genes cloned in maize that can be used in molecular breeding

Method used

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  • Gene MS33 related to maize male nuclear sterility and application thereof in cross breeding
  • Gene MS33 related to maize male nuclear sterility and application thereof in cross breeding
  • Gene MS33 related to maize male nuclear sterility and application thereof in cross breeding

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Example 1. Map-based cloning of the gene MS33 related to maize male sterility

[0061] 1. Phenotype identification of maize ms33 mutant

[0062] The maize male sterile 33 (ms33) mutant was discovered in 1999, and there are 8 alleles of the ms33 mutant preserved in the U.S. Maize Genetics Cooperation Stock Center public mutant bank at present, and the inventor of the present invention applied for The ms33 mutant allele ms33-6019 (ie maize mutant ms33-6019) was obtained and phenotyped. turn out. Compared with wild-type maize plants (inbred line Zheng 58), the ms33-6019 mutant showed no significant difference in pollen abortion and vegetative growth stage ( figure 1 Middle A), but when the ms33-6019 mutant plant entered the pollination stage, the anthers could not be spit out from the florets. Through microdissection and fertility identification, it was found that the ms33-6019 mutant anthers were severely atrophied, and the anthers were treated with 1% I 2 -KI soluti...

Embodiment 2

[0075] Example 2, Maize ms33 Mutant MS33 Gene Functional Complementation Experiment

[0076] In order to further prove and confirm that the ms33 mutant phenotype is caused by the mutation of the MS33 gene, the inventors of the present invention conducted another ms33 allelic mutant line ms33-6024 (that is, the maize mutant ms33-6024, which has the same male sterility as ms33-6019) traits) genome sequencing, and found that compared with the wild-type maize plant—inbred line Zheng 58, the sequence difference of the ms33-6024 mutant was only the insertion of 2 bases at the 507 bp base in the coding region of the MS33 gene, which resulted in migration code mutation, premature termination of translation ( image 3 Middle B).

[0077] At the same time, the inventors of the present invention screened a new ms33 mutant allele sa1 in the self-developed MuDR mutant library. The heterozygous fertile plants (+ / ms33) from the ms33-6019 allele were pollinated to the sa1 homozygous sterile...

Embodiment 3

[0079] Embodiment 3, the expression analysis of MS33 gene in each organ

[0080] In order to further study the function of MS33 gene in detail, the inventors of the present invention extracted different organs (roots, stems, leaves and anthers of different developmental stages of wild-type maize plant (inbred line Zheng 58), i.e. sex mother cell stage, quarter Somatic, mononucleate, dinucleate, and mature pollen) RNAs were reverse-transcribed to obtain cDNA, and the expression of MS33 gene in different tissues was detected by fluorescent quantitative PCR. Among them, the primers used to detect MS33 gene are MS33-F and MS33-R; maize Aciton is used as an internal reference, and the detection primers are Actin-F and Actin-R.

[0081] MS33-F: 5'-TCTGCGTCACTGCCATGC-3';

[0082] MS33-R: 5'-CGTCGGAGCTCACCACG-3'.

[0083] Actin-F: 5'-GAGATGCCTGATGGTCAGGTCA-3';

[0084] Actin-R: 5'-AGTTGTACGTGGCCTCATGGAC-3'.

[0085] The results showed that the expression of MS33 gene was higher in...

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Abstract

The invention discloses a gene MS33 related to maize male nuclear sterility and application thereof in cross breeding. The invention provides MS33 protein related to maize male nuclear sterility, which is shown in (a) or (b): (a) protein composed of an amino acid sequence shown in a sequence 1 in a sequence table; or (b) protein which is derived from the sequence 1 by substituting and / or deleting and / or adding one or more amino acid residues on the amino acid sequence of the sequence 1, and related to plant male fertility. MS33 gene mutation causes maize complete male sterility, characters are complete, and environmental influence is avoided. The gene has application potential in maize hybrid seeds.

Description

technical field [0001] The invention belongs to the field of plant genetic engineering and relates to a gene MS33 related to maize male sterility and its application in hybrid breeding. Background technique [0002] Corn is my country's largest food crop, and my country's corn production and consumption ranks second in the world. Therefore, the stable increase of corn production plays a very important role in maintaining domestic food security. In recent years, with the rapid development of downstream industries, the demand for corn is increasing day by day, and the domestic demand for corn needs to be regulated by imports. Maize production is under increasing pressure. [0003] Currently, maize seed production relies extensively on hybrid seed production. Traditional hybrid seed production requires strict artificial detasseling and controlled pollination to ensure the purity of commercial seeds, which consumes a lot of labor costs and land resources. The impact of this ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/10C12N15/54C12N15/82A01H5/00
Inventor 金危危张磊罗红兵陈晓阳赵悦
Owner CHINA AGRI UNIV
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