Mapping and Preliminary Analysis of ABORTED MICROSPORES (AMS) as the Candidate Gene Underlying the Male Sterility (MS-5) Mutant in Melon (Cucumis melo L.)
Mapping and Preliminary Analysis of ABORTED MICROSPORES (AMS) as the Candidate Gene Underlying the Male Sterility (MS-5) Mutant in Melon (Cucumis melo L.)
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甜瓜（Cucumis melo L.）雄性不育（MS-5）突变体候选基因败育小孢子（AMS）的定位和初步分析
DOI：
10.3389/fpls.2017.00902
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发表时间：
2017-05-30
影响因子：
5.6
通讯作者：
Luan, Feishi
中科院分区：
文献类型：
--
作者：
Sheng, Yunyan;Wang, Yudan;Luan, Feishi
甜瓜是世界上重要的农业和经济蔬菜作物。遗传性雄性不育突变体ms-5具有控制雄性不育种质的隐性核基因。雄性不育可降低甜瓜F1代制种成本，但授粉前应去除杂合可育植株。本研究采用群体分离分析结合特异长度扩增片段测序法对水稻单核雄性不育隐性基因进行了定位。基于亲本系重测序数据和252个F2个体的经典作图，鉴定了位于支架000048上的9号染色体上的30-kb候选区域，其跨度为2，522，791至2，555，104 bp，并通过切割扩增多态性序列标记进一步证实。基因预测表明，6个注释的基因存在于30-kb的候选区域。定量RT-PCR结果表明，不育系（ms-5）和可育系（HM 1 -1）在2-mm（四分体）和5-mm（第一次花粉有丝分裂）时期的AMS基因表达量存在显著差异，AMS候选基因转录因子也存在负调控。AMS转录因子的测序和聚类分析揭示了亲本系之间的五个单核苷酸多态性。本研究结果提示AMS转录因子可能是甜瓜单核雄性不育的候选基因。本研究结果将有助于育种者在播种时识别雄性不育和可育植株，作为分子标记辅助选择方法，这将降低种子生产成本，提高甜瓜雄性不育系的利用率。
Melon is an important agricultural and economic vegetable crop worldwide. The genetic male sterility mutant (ms-5) has a recessive nuclear gene that controls the male sterility germplasm. Male sterility could reduce the cost of F1 seed production in melon, but heterozygous fertile plants should be removed before pollination. In this study, bulked segregant analysis combined with specific length amplified fragment sequencing was applied to map the single nuclear male sterility recessive gene. A 30-kb candidate region on chromosome 9 located on scaffold 000048 and spanning 2,522,791 to 2,555,104 bp was identified and further confirmed by cleavage amplified polymorphic sequence markers based on parental line resequencing data and classical mapping of 252 F2 individuals. Gene prediction indicated that six annotated genes are present in the 30-kb candidate region. Quantitative RT-PCR revealed significant differences in the expression level of the LOC103498166 ABORTED MICROSPORES (AMS) gene in male-sterile lines (ms-5) and male-fertile (HM1-1) lines during the 2-mm (tetrad) and 5-mm (the first pollen mitosis) periods, and negative regulation of the AMS candidate gene transcription factor was also detected. Sequencing and cluster analysis of the AMS transcription factor revealed five single-nucleotide polymorphisms between the parental lines. The data presented herein suggest that the AMS transcription factor is a possible candidate gene for single nuclear male sterility in melon. The results of this study will help breeders to identify male-sterile and -fertile plants at seeding as marker-assisted selection methods, which would reduce the cost of seed production and improve the use of male-sterile lines in melon.