Gene Mapping and Genetic Analysis of Male Sterile Mutant x50 in Maize

doi:10.7668/hbnxb.20193481

Abstract

Abstract:

In order to explore the germplasm resources of male sterile line and identify the male fertility genes, which would provide the basic materials for the maize seed production by the male sterile line. Using the male sterile mutant x50 as the experimental material,the male sterile phenotype of x50 was studied,and the F₁ and F₂ populations of x50 and inbred line Mo17 were constructed to determine the genetic pattern of the male sterile trait in x50.With the F₂ population,the male fertility gene X50 was identified by the map-based cloning.Furthermore,the candidate gene was confirmed by the allelism test.The results showed that compared with the wild type,the x50 anthers did not emerge from the glume,and was smaller and wilted,and no mature pollen grains were formed.All F₁ plants were fertile,and F₂ plants displayed 3∶1 segregation ratio between fertile and sterile plants,indicating that the monofactorial recessive inheritance of x50.The gene X50 was mapped to the interval from the molecular marker 2-4901 to 2-4963 on chromosome 2 with a physical range of 237.42 to 241.39 Mb by map-based cloning.Candidate gene analysis found that the male fertility gene ZmMs33 was located in the mapping region.In addition,the test crosses of the ms33 mutants(ms33-6029 and ms33-6052)and heterozygous +/x50 revealed 1∶1 segregation ratio between fertile and sterile plants.These results suggested that x50 was an allele mutant of ZmMs33 gene.The identification of the male sterile mutant x50 provided germplasm resources for maize hybrid seed production and functional study of ZmMs33 gene.

Key words: Maize, Male sterile gene X50, Gene mapping, Anther, Allelism test

GUO Yaoqing, SUN Xiaojing, LIAN Yujie, CHEN Hui, SUN Huayue, ZHANG Xuehai, TANG Jihua, CHEN Xiaoyang. Gene Mapping and Genetic Analysis of Male Sterile Mutant x50 in Maize[J]. Acta Agriculturae Boreali-Sinica, 2023, 38(1): 17-22. doi: 10.7668/hbnxb.20193481.

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Figures/Tables 7

Fig.1 Phenotypic comparison between the wild type and x50 mutant A.The plants of the wild type and x50;B.The tassels of the wild type and x50;C.The spikelets of the wild type and x50;D,E.The mature pollen grains of the wild type(D)and x50(E).

Tab.1 Phenotypic analysis of male fertility in F₂ population

时间和地点 Time and location	可育株数 No. of fertile plants	不育株数 No. of sterile plants	χ² (3∶1)
2019年,海口 2019,Haikou	153	48	0.13
2019年,原阳 2019,Yuanyang	190	64	0.01

Fig.2 The identification of linked markers with X50 gene

Tab.2 Primers used in the gene mapping of X50

引物名称 Primer name	引物序列(5'—3') Primer sequence(5'—3')
2-4791-F	GTCGCTGCTGCCCATGTTATTC
2-4791-R	TGCTGTGCCCGTGATCTCATC
2-4901-F	AACTCCAATCCGCCTCGACTG
2-4901-R	GTAGCCGTATCCTCGTCGTCTG
2-4963-F	TGCTGGTTAATCTGTGCCTTGC
2-4963-R	ACCTGTGTTGCCAAACTTGTTG

Fig.3 The gene mapping of X50

Fig.4 Allelism test of x50 and ms33 mutant A.The tassels of x50/ms33-6029 and wild type derived from the cross of ms33-6029 and heterozygous+/x50;B and C.The mature pollen grains of x50/ms33-6029 and wild type derived from the cross of ms33-6029 and heterozygous+/x50.

Tab.3 Fertility phenotype of the crosses of ms33 mutant and heterozygous +/x50

组合 Combinations	不育株数 No. of sterile plants	可育株数 No. of fertile plants	χ² (1∶1)
ms33-6029×+/x50	34	28	0.58
ms33-6052×+/x50	16	17	0.03

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