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Acta Agronomica Sinica ›› 2025, Vol. 51 ›› Issue (10): 2595-2604.doi: 10.3724/SP.J.1006.2025.53023

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• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

Different mutations in ZmMS13 generate the gms1 and yems1166 multiple allele maize male-sterile lines

ZHAO Zhi-Wen(), CHEN Hui(), LIAN Yu-Jie, LU Han, CAO Xu-Dong, WANG Fan, YU Meng-Fan, ZHANG Zhan-Hui, TANG Ji-Hua, CHEN Xiao-Yang()   

  1. College of Agronomy, Henan Agricultural University / State Key Laboratory of High-Efficiency Production of Wheat-Maize Double Cropping, Zhengzhou 450046, Henan, China
  • Received:2025-04-15 Accepted:2025-07-09 Online:2025-10-12 Published:2025-07-15
  • Contact: *E-mail: cxy759020@henau.edu.cn E-mail:zzw16692520062@163.com;17339301889@163.com;cxy759020@henau.edu.cn
  • About author:First author contact:**Contributed equally to this work
  • Supported by:
    Joint Foundation for Science and Technology Research and Development Program of Henan Province (Cultivation of Superior Disciplines)(242301420131);Zhengzhou Research and Development Project of Open Bidding for Selecting the Best Candidates(2023JBGS013)

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Abstract:

The development and utilization of maize male-sterile lines is an effective strategy to reduce the cost of hybrid seed production and improve seed purity. In this study, two maize male-sterile mutants, gms1 and yems1166, were identified. Phenotypic analysis revealed that both mutants exhibited abnormal anther cuticle structures, microspore degradation at the late uninucleate stage, and complete absence of mature pollen grains. Genetic analysis indicated that male sterility in both mutants is controlled by a single recessive nuclear gene. Using a positional cloning approach, the GMS1 and YEMS1166 loci were mapped to intervals of 23.52-26.09 Mb and 24.86-30.95 Mb on chromosome 5, respectively, regions containing the previously reported male fertility gene ZmMS13, which encodes the ABCG transporter ZmABCG2a. Gene sequencing revealed that gms1 carries a single base substitution (TCA>TGA) in the fifth exon of ZmMS13, resulting in a premature stop codon. The yems1166 mutant harbors an 8-bp deletion in the first exon of the same gene, also leading to a premature stop codon. Allelic tests confirmed that gms1 and yems1166 are novel allelic variants of ZmMS13. These findings identify two new allelic mutants of ZmMS13 and provide valuable germplasm resources for the development of male-sterile lines in maize hybrid seed production.

Key words: maize, male sterility, anther cuticle, ms13, ABCG transporter

Table 1

Primer sequences used for gene mapping"

引物名称
Primer name		 正向引物序列
Forward primer sequence (5′-3′)		 反向引物序列
Reverse primer sequence (5′-3′)
S08836 ACCAGTCCGCCACTACTATTCG CCGCTCCATCCGTCCTGTC
S08958 GGTGCCCGTTACATCAGATCAG GTGTGACCGTGTGAGTGGAATC
S08994 TGCTTGCCTTCTCCTGTTTGAC GCTCGGTTCACGCCTTCAC
S09011 TCCGTTCGGAATCACTTGTTGG TTCAGGAGGGAGCGGAGTTG
S09069 GCAACGTGCATGTGTGTG GTCAACGGCCAGCCCTAC
S09079 GCCAGCGTCACCGTCGTC TGTCGTGCGGAAGCCTGAAG
S09092 TCCTTATCCCTGCCAAGAAACG AGAGTGCGGAGAGTGAGACC
S09100 CCTTTCTGAATGGAGGGAGTAC CGTCTGGCTTCTTTTCATGTTG
S09128 TTAGGATCAGCAAGGCATAGGG GAGCGTTGTCAGGTAAGTTTGG
S09130 TGACTCATTCAGCAGCCTGGTG CCGTTGACTGGTCCACCTGTAC
S09138 TACCCGTCCCAATTCCCAACC AAGCAGCGAACCAGCACTTG
S09144 CCTTGGCTGTCTCCAGTAGTCC CCTGCTGGTCCTCTGGTTGTG
S09146 TGCCTGCCTATATCTTCGACAC TCAATTCCTCGGTAGCGTCATC
S09154 GCTTCTTGTCGGAAATCGTCAC TTTGGCAGGTTCAGCTAATTGC
S09160 GACACCATGCCTGTGACTGAG AAAGCCGTTTCTTGCCCTCTG
S09172 GCCGTGTTGGAATGATGGTC TCTTCAGCTAGAGAGGTGGTAC
S09220 CAACAACACAACGCCACAAC TGATGCCATGATGACCTGTATG

Fig. 1

Phenotypic analysis of wild type (WT) and gms1 at the flowering stage A: wild-type and gms1 plant, scale bar=30 cm; B: wild type and gms1 tassel, scale bar=1 cm; C: wild type and gms1 spikelet, scale bar=2 mm; D: wild type mature pollen grains, scale bar=100 μm; E: gms1 mature pollen grains, scale bar=100 μm."

Fig. 2

Comparison of anther morphology at the different stages between wild type (WT) and gms1 mutant S8b: the tetrad stage; S9: the early uninucleate stage; S10: the late uninucleate stage; S11: the binucleate stage; S13: the mature pollen grain stage. Scale bar = 1 mm."

Fig. 3

Observation of microspore development at different stages A, B: tetrad stage; C, D: early uninucleate stage; E, F: late uninucleate stage; H, I: binucleate stage; J, K: mature pollen grain stage. WT: wild type. Scale bar = 100 μm."

Fig. 4

Structural analysis of the anther cuticle at the mature pollen grain stage A, B: anthers, scale bar=1 mm; C, D: outer surface of anther, scale bar=25 μm; E, F: inner surface of anther, scale bar=5 μm. WT: wild type."

Table 2

Segregation ratio of fertile plants and sterile plants in the F2 population"

时间
Time		 地点
Location		 F2分离群体
F2 population		 雄性可育株
Male fertile plants		 雄性不育株
Male sterile plants		 χ2
2022年春Spring in 2022 原阳Yuanyang gms1×B73 108 26 2.328
2022年夏Summer in 2022 原阳Yuanyang gms1×B73 46 15 0.011

Fig. 5

Gene mapping of GMS1"

Fig. 6

Gene mapping of YEMS1166"

Table 3

Segregation ratio of fertile plants and sterile plants in the population"

时间
Time		 地点
Location		 群体
Population		 雄性可育株
Male fertile plants		 雄性不育株
Male sterile plants		 χ2
2023 海南Hainan gms1×(yems1166×B73) 31 28 0.508
yems1166×(gms1×B73) 37 38 0.027
2023 原阳Yuanyang gms1×(yems1166×B73) 35 32 0.400
yems1166 ×(gms1×B73) 27 24 0.467

Fig. 7

Gene sequencing of ZmMS13"

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