作物杂志,2026, 第5期: 107–114 doi: 10.16035/j.issn.1001-7283.2026.05.013

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玉米黄化突变基因ZmET10的精细定位

虞思佳1,2(), 腾峰2, 童少华3, 王明成3, 叶文举1,2, 刘丹2, 贺正华2, 张士龙2, 孙霄鹏2, 李曼菲1, 杜何为1(), 贾海涛2()   

  1. 1 长江大学生命科学学院, 434025, 湖北荆州
    2 湖北省农业科学院粮食作物研究所, 430064, 湖北武汉
    3 谷城县农业科学研究所, 441700, 湖北襄阳
  • 收稿日期:2025-08-04 修回日期:2025-09-25 出版日期:2026-10-15 发布日期:2026-10-22
  • 通讯作者: 杜何为,研究方向为玉米遗传育种,E-mail:200457@yangtzeu.edu.cn;贾海涛为共同通信作者,研究方向为玉米遗传育种,E-mail:htjia@hbaas.com
  • 作者简介:虞思佳,研究方向为玉米种质资源创新,E-mail:17816918221@163.com
  • 基金资助:
    湖北省农业科学院粮食作物研究所自主创新基金(2022ZZCX003);湖北省自然科学基金创新群体项目(2022CFA030)

Fine Mapping of the Etiolation Mutant Gene ZmET10 in Maize

Yu Sijia1,2(), Teng Feng2, Tong Shaohua3, Wang Mingcheng3, Ye Wenju1,2, Liu Dan2, He Zhenghua2, Zhang Shilong2, Sun Xiaopeng2, Li Manfei1, Du Hewei1(), Jia Haitao2()   

  1. 1 College of Life Science, Yangtze University, Jingzhou 434025, Hubei, China
    2 Food Crops Institute, Hubei Academy of Agricultural Sciences, Wuhan 430064, Hubei, China
    3 Gucheng Institute of Agricultural Sciences, Xiangyang 441700, Hubei, China
  • Received:2025-08-04 Revised:2025-09-25 Online:2026-10-15 Published:2026-10-22

摘要:

叶色突变体的表型变异可归因于叶绿体结构的发育缺陷与叶绿素代谢途径的紊乱。为探究玉米叶色变异的分子机制,从甲基磺酸乙酯(EMS)诱导的B73突变体库中鉴定出1份能稳定遗传的黄化材料et10。通过遗传学分析,证实该突变表型由1对隐性核基因ZmET10控制。利用集群分离分析结合转录组测序(BSR-Seq)策略,将相关位点锚定于第10号染色体长约19 Mb的区段。基于该区域设计并筛选了10个SSR和5个InDel多态性标记,对1628株BC1F1群体中显现黄化表型的个体进行基因型分析。最终将ZmET10定位于第10号染色体标记indel-62和indel-69之间约30.2 kb的区间内。该区间内含有7个候选基因,其中Zm00001d025868编码细胞分裂素核苷5′-单磷酸磷酸核糖水解酶,其功能异常可能导致活性细胞分裂素水平降低,进而引起植物的黄化表型,推测该基因为ZmET10的关键候选基因。

关键词: 玉米, 黄化突变基因ZmET10, BSR-Seq, 精细定位

Abstract:

Phenotypic variation in leaf-color mutants can be attributed to developmental defects in chloroplast structure and disturbed chlorophyll metabolism. To explore the molecular mechanism of maize leaf-color variation, a stably inherited etiolation mutant et10 was identified from an ethyl methanesulfonate (EMS)- mutagenized library of maize inbred line B73. Genetic analysis revealed that the mutant phenotype was controlled by a single recessive nuclear gene, designated ZmET10. Using BSR-Seq, the target gene was primarily mapped to a 19 Mb region on chromosome 10. Furthermore, ten SSR and five InDel markers were developed in this region. By genotyping 1628 recessive individuals from a BC1F1 population, ZmET10 was fine-mapped to a 30.2 kb interval between markers indel-62 and indel-69. This interval harbors 7 candidate genes. Among them, Zm00001d025868 encodes cytokinin riboside 5'-monophosphate phosphoribohydrolase, whose loss of function may reduce active cytokinin levels and lead to the etiolation phenotype. Thus, Zm00001d025868 is proposed as the candidate gene for ZmET10.

Key words: Maize (Zea mays), Etiolation mutant gene ZmET10, BSR-Seq, Fine-mapping

表1

ZmET10基因定位相关引物

引物Primer F-序列(5-3′)F-sequence (5-3′) R-序列(5-3′)R-sequence (5-3′)
umc1739 CAAAAAGCTTTAGTTTTTACAGGCAGA GCGTGTAATTTGCTCCTTCTTCTT
bnlg1712 CTCAGGCTTCACGTGGGTTT GTTACACTCCCCTGCCAAAA
umc1911 CCCGGAAGCTGTTTTCTTTTTAAT CGATCTGGAACGTAAGGATCATCT
umc1453 AATACCAAGCTGCACTCAGAAACC CGTCAAATCCAGCCTAAGCATC
umc1330 AGCAAAGAAGCCAAAGAGCAACT GTCACCACCGTCTGCTGGTA
umc1930 TCTTCTCCAAGTGTGTTAATGCCC CACACAGTGAGTCGTTTCTTTCGT
bnlg1074 CATGCTAATAGCCTACCGGG TTTCCCCCTGATTCGTTATG
umc1506 AAAAGAAACATGTTCAGTCGAGCG ATAAAGGTTGGCAAAACGTAGCCT
umc2221 ATAATGCTACACTTTTGCGGGGTT CAGGCGGATAAGAATGACGAGTAG
bnlg2190 TCCTCCTTCATCCCCTTCTT CCCAGTATCATTGCCCAATC
indel-48 GGCATGCTCCATTAGATCGT CCGAGAATAGCCACTCAAGC
indel-61 TAGACCTTTTGCCGAGTGCT CGCGCGATCTCTTATCATTA
indel-62 GCTTCCAAGCATCGTAATCC GCCTTCGAGCCAACAATAAC
indel-64 ATGCCGCAGTCGTCGCCGAGC CACTCCTACAAGCAAATAGATT
indel-69 TGCATCGTCATAATCTGAAGC CTTTCAAACTCCTACCGTGCT

图1

野生型B73与玉米黄化突变体et10田间表型调查 (a) 苗期表型;(b) 苗期叶片;(c) 成株期;(d) 茎秆;(e) 雄穗分枝;(f) 未成熟果穗。

表2

et10及其亲本B73农艺性状考察分析

性状
Trait
B73 et10 P值
P-value
株高Plant height (cm) 175.69±9.51 125.63±12.86 4.08E-21**
穗位高Ear height (cm) 66.75±5.56 45.62±9.57 2.16E-09**
雄穗分枝数
Number of tassel branches
4.69±0.93 4.43±1.24 0.24
单株叶片数
Number of leaves per plant
13.56±0.78 12.33±0.54 1.51E-08**
叶长Length of leaf (cm) 73.31±4.68 65.94±4.17 3.51E-06**
叶宽Width of leaf (cm) 8.18±0.67 8.54±0.70 0.05*
穗长Ear length (cm) 9.25±1.10 7.83±1.11 0.0006**
穗重Ear weight (g) 73.01±8.04 38.38±7.17 4.49E-10**
百粒重100-grain weight (g) 25.91±1.37 18.87±2.03 2.71E-09**
抽雄天数Days to tasseling (d) 75.52±1.62 78.69±1.57 2.54E-07**
散粉天数Days to shedding (d) 77.71±1.43 80.59±1.51 3.17E-11**
吐丝天数Days to silking (d) 78.54±1.32 82.97±1.41 4.12E-16**

表3

2个杂交组合F1和F2群体的分离

组合
Combination
群体Population 比率
Ratio
卡方
χ2
P值
P-value
F1 F2
WT et
B73×et10 绿色 217 74 2.9 0.31 0.65
Mo17×et10 绿色 206 62 3.3 0.32 0.53

图2

野生型B73与突变体et10色素含量 “**”表示在P < 0.01水平差异显著。

图3

透射电镜下的野生型B73和突变体et10叶绿体结构

图4

BSR-Seq分析揭示黄化突变基因ZmET10的遗传定位

图5

ZmET10的精细定位 WT:野生型;Mu:突变体。

表4

精细定位区间内的7个基因

基因ID Gene ID 位置Locus 功能描述Functional description
Zm00001d025863 131891540~131892317 光依赖性短下胚轴蛋白
Zm00001d025864 131914989~131917182 MYB结构域蛋白
Zm00001d023172 131919025~131919626 未知功能蛋白
Zm00001d025865 131993604~132007438 肌醇磷酸神经酰胺葡萄糖醛酸基转移酶1
Zm00001d025866 132009297~132012863 核小核糖核蛋白家族蛋白
Zm00001d025867 132084448~132087460 未知功能蛋白
Zm00001d025868 132094786~132098109 细胞分裂素核苷5′-单磷酸磷酸核糖水解酶
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