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Acta Agronomica Sinica ›› 2020, Vol. 46 ›› Issue (12): 1991-1996.doi: 10.3724/SP.J.1006.2020.03025

• RESEARCH NOTES • Previous Articles     Next Articles

Genetic analysis and characterization of male sterile mutant mi-ms-3 in maize

TIAN Shi-Ke(), QIN Xin-Er, ZHANG Wen-Liang, DONG Xue, DAI Ming-Qiu, YUE Bing*()   

  1. National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, Wuhan 430070, Hubei, China
  • Received:2020-05-07 Accepted:2020-08-19 Online:2020-08-31 Published:2020-08-31
  • Contact: YUE Bing E-mail:tianshike1996@163.com;yuebing@mail.hzau.edu.cn
  • Supported by:
    National Key Research and Development Program of China(2016YFD0100804)

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Abstract:

Maize is one of the best crops in the utilization of heterosis. Male sterile lines are important germplasms for the hybrids production. A male sterile mutant named mi-ms-3 was obtained by screening in a mutator insertion library. The number of male anthers in tassel decreased and not exserted. There were few anthers with only two pollen sacs in the mutant tassels, and some of the anthers were degenerated to membranous and formed filaments at their ends. Although pollens in the anthers could be stained by I2-KI, pollen shedding was abnormal and the number of pollen grains decreased. The number of silks in the ear of the mutant increased, and there was a sterile grain on both sides of the maturated kernel. Fertility of F1 plants, which were obtained by hybridization between mi-ms-3 and maize inbred Mo17, was normal. Genetic analysis of F2 population showed that the mutant phenotype was controlled by a recessive gene. The candidate gene was preliminarily mapped on the long arm of chromosome 3 by BSA and it was located between a SSR marker and an Indel marker with a distance of 1.5 cM. There are 21 candidate genes in this region. It was finally found that the insertion mutation of Mu transposon occurred at 30 bp upstream of the coding region of zm00001d042618 (zmm16) by transponson tagging and sequencing analysis. The results showed that mi-ms-3 was a new allele of sts1, which caused by a single base mutation in the coding region. RT-PCR analysis indicated that the expression of zmm16 in the mutant was decreased. The identification of the new allelic mutant of sts1 in this study would provide new materials for the study of flower development and hybrid seed production.

Key words: maize, male sterile, gene mapping, genetic analysis

Table 1

Primers for genetic mapping and gene expression analysis"

引物名称
Primer name		 正向引物
Forward sequence (5′-3′)		 反向引物
Reverse sequence (5′-3′)		 染色体位置
Chromosome position
umc1973 CAGGCAGAAAAGGAACGGAAC GTGCGAGAGAAGATGGATGATTG 3.05
umc1027 AACTCTGTCTCCGTCACCGTGT GACCTCATCTCGGTGGAAATTG 3.06
S-1 CCGTCATCGACATTTTACTCAG CACAAGAGTTGGTTCGCTG 3.06
S-2 CCTTGTACCCGCTCATCCAT CTCCTACCGACCTGCCTG 3.06
S-3 GTGTTACAAGAACTCAGCGT AGCCTTCGTTCAGTTTCGGT 3.06
S-4 TGACTGGCTTTCGTCCTACT CACTCTCCAGCTGCTTCTCT 3.06
S-5 ACAATTAGCTTAAGAACGC AGCGGTGTTCATGGTACTCA 3.06
S-6 TGGCTACTAGCTAGCTGTGCT CTAAACCTTCACGCACAGCC 3.06
S-7 CCAACTGAGCCGAATCGTTC ATCATCCCATATCCCACGCC 3.06
S-8 GTGGCCCATATGGTCTCAAC CGGCCAGTCGATTAAAGCTC 3.06
S-9 CGACGAAGAACTGACTTGACC TTGCTTGATCATTCCTCCGC 3.06
S-10 GTTTGGGCCGCATTCGAGAGA TATTCTTGCCTCTCCCACCG 3.06
TIR-1 CGGCCTCCATTTCGTCGAATCCCTT
42618 TGCCGGTACTGTGACTGTTC CCTGAGCTCGATCTGCATGG 3.06
42618-2 TCTTTACTCCTCCCCTCCCA CTTCTTGAGGATCCCGTTGC 3.06
q42618 GCCTTAGTGCAGAGATTGACC GCCTTTCCCAGTGCTCCA 3.06

Fig. 1

Phenotypic of the mi-ms-3 and wild type (WT) A: the tassel of WT(wild type) (left) and mi-ms-3 (right), mi-ms-3 anther not exserted; B1-B2: transverse sections of the entire anther of WT and mi-ms-3 in the late microspore developmental stage, mi-ms-3 anthers with only two pollen sacs; C1-C4: the spikelet of mi-ms-3 contain 0-3 anthers respectively; D1-D2: pollen grains of WT and mi-ms-3; E1: the ear of mi-ms-3 (left) and WT (right), the number of silks in the ear of the mi-ms-3 increased; E2: the kernel of mi-ms-3 (left) and WT (right), mi-ms-3 each seed corresponds to multiple silks."

Table 2

Genotype and phenotype of the important recombinant individuals"

umc1973 S-1 S-2 S-3 S-4 S-5 S-6 S-7 S-8 S-9 S-10 umc1027 表型Phenotype
2-2 H H H H A A A A A A A A 突变体Mutant
11-5 H H H H H A A A A A A A 突变体Mutant
9-3 H H H H H H H A A A A A 突变体Mutant
14-8 A A A A A A A A A A A H 突变体Mutant
5-4 H H H H H H H H H H H A 野生型Wild type
20-9 A H H H H H H H H H H H 野生型Wild type

Fig. 2

Identification of insertion mutant of mu transposon 1-2: heterozygous plants; 3-5: homozygous wild type plants; 6: marker; 7-14: homozygous mutant plants; 15: B73; 16: Mo17."

Fig. 3

Sequence analysis of zmm16 Mu13 transposon occurred at 30 bp upstream of ATG of zmm16."

Fig. 4

Prediction and analysis of promoter elements of zmm16"

Fig. 5

Expression level of zmm16 The expression of zmm16 in mi-ms-3 decreased compared with wild type."

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