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作物学报 ›› 2020, Vol. 46 ›› Issue (12): 1831-1838.doi: 10.3724/SP.J.1006.2020.03017

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

玉米籽粒突变体dek101的表型分析和精细定位

宋欣冉1(), 胡书婷2(), 张凯2, 崔则瑾2, 李建生2, 杨小红2, 白光红1,*()   

  1. 1新疆农业大学, 新疆乌鲁木齐830052
    2中国农业大学 / 国家玉米改良中心 / 农业农村部玉米生物学重点实验室, 北京100193
  • 收稿日期:2020-03-17 接受日期:2020-07-02 出版日期:2020-12-12 网络出版日期:2020-11-25
  • 通讯作者: 白光红
  • 基金资助:
    国家自然科学基金项目(31421005)

Phenotypic analysis and fine mapping of dek101 in maize

Xin-Ran SONG1(), Shu-Ting HU2(), Kai ZHANG2, Ze-Jin CUI2, Jian-Sheng LI2, Xiao-Hong YANG2, Guang-Hong BAI1,*()   

  1. 1Xinjiang Agricultural University, Urumqi 830052, Xinjiang, China
    2National Maize Improvement Center of China / Key Laboratory of Maize Biology of Ministry of Agriculture and Rural Affairs / China Agricultural University, Beijing 100193, China
  • Received:2020-03-17 Accepted:2020-07-02 Published:2020-12-12 Published online:2020-11-25
  • Contact: Guang-Hong BAI
  • Supported by:
    National Natural Science Foundation of China(31421005)

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摘要：

籽粒作为玉米储藏器官, 其发育程度和物质储存直接影响玉米的产量和品质。本研究在玉米双单倍体系选育过程中发现可稳定遗传的籽粒缺陷突变体, 命名为defective kernel 101 (dek101)。该突变体籽粒皱缩, 粒重显著降低, 胚致死, 胚乳发育缺陷, 不能成苗。在授粉后12 d, dek101开始出现明显的发育异常, 授粉后21 d籽粒鲜重、干重、体积不再增加。扫描电镜观察发现, 与野生型相比, dek101淀粉粒显著变小。遗传分析证实该突变性状受隐性单基因控制。利用441个F2单株和1648个F3单株, 将该基因定位在1号染色体的标记IDP2182和IDP4600之间, 物理区间约300 kb, 共有5个预测基因。这些结果为挖掘与玉米籽粒发育有关的功能基因, 解析籽粒发育机制奠定了基础。

关键词: 玉米, 籽粒缺陷突变体, 精细定位

Abstract:

As the storage organ of maize, kernel development and accumulation of storage production directly determines maize yield and quality. In this study, a stable defective kernel mutant, named as defective kernel 101 (dek101), was identified during the development of double haploid (DH) lines in maize. The dek101 kernels displayed severely shrunk kernel appearance, significantly reduced kernel weight, lethal embryo, defective endosperm and were incapable of germinating. The dek101 showed obvious developmental abnormalities at 12 days after pollination (DAP). The fresh weight, dry weight and volume of the kernels were no longer increased after 21 DAP. Scanning electron microscopy (SEM) observation revealed that the starch granules of dek101 were significantly smaller compared with wild-type kernels. Genetic analysis demonstrated that the mutant trait was controlled by a recessive single gene. Using 441 F2 individuals and 1648 F3 individuals, dek101 was narrowed down to a genomic region of about 300 kb between the InDel marker IDP2182 and IDP4600 on chromosome 1, which contains five predicted genes. These results laid the foundation for mining functional genes related to maize kernel development and deciphering the mechanism of grain development.

Key words: maize, defective-kernel mutant, fine mapping

表1

在亲本间有多态性InDel引物"

引物
Primer		 位置
Location (Mb)		 序列
Primer sequence (5′-3′)		 产物
Product (bp)
IDP112 42.1 F: AGGAAGCTGTATCCCACACG; R: TCATGGGTTTCTTCTTTGCG 624
IDP8266 43.3 F: GTTGTTGTGCTCCAGAGAAGG; R: TACGTTGCCTATCATTGCCC 369
IDP101 45.1 F: CTATCCCGTTCGTGTTCA; R: CCCTGCGTTGTCTTTCTC 261
IDP643 46.2 F: ACCCTCATCTTCAGCAGTCG; R: GGTGAAACGGCAGTACAAGG 838
IDP2182 47.1 F: GGAATGTGTACACGGCAGGT; R: ACAGCAATCGGAGCAGTGTT 225
IDP4600 47.4 F: CACTTCGACGAGGGGTTCAT; R: GTAACCACCTACCCACAA 263
IDP8401 49.3 F: TAGGCACTGTTTGTTTCA; R: CTAGGGTTATGTGGCATT 324
IDP78 49.3 F: CGGTTGTGACTGCTATGT; R: TGTGGCATTTATTGTTCAT 342

图1

籽粒发育动态及授粉后不同时期野生型籽粒和突变型籽粒对比 A: 杂合果穗; B: 30 DAP时突变型籽粒和野生型籽粒对比; C: 授粉后不同时期野生型与突变型籽粒发育动态; D~E: 50颗野生型和突变型籽粒不同时期干重、鲜重的变化; F: 20颗野生型和突变型籽粒不同时期体积的变化。WT: 野生型籽粒; dek101: 突变体籽粒。A~C图标尺为1 cm。**, 突变型籽粒与野生型相比在统计学上有极显著差异 (P < 0.01)。DAP: 授粉后天数。"

图2

野生型与dek101籽粒淀粉粒扫描电镜观察 A~C: 授粉后15、21和27 d野生型籽粒扫描电镜观察; D~F: 授粉后15、21和27 d突变型籽粒扫描电镜观察。A~F图标尺: 5 μm。"

表2

F2果穗突变型籽粒的分离比例"

果穗
Ear		 野生型籽粒数
Wild-type		 突变型籽粒数
Mutant		 χ2
1 268 86 0.06
2 292 103 0.19
3 252 85 0.001
4 241 80 0.001
总计Total 1053 354 0.019

图3

dek101的精细定位 n: F2群体大小以及F3群体数目; 黑色竖线代表引物位置; 标记下方的红色数字代表重组单株数目; 矩形框代表5个重组类型的基因型以及对应的籽粒表型, 黑色矩形代表B73基因型, 灰色矩形代表杂合基因型。"

表3

dek101定位区间的候选基因注释"

基因位点
Gene locus		 基因注释
Gene annotation		 染色体
Chr.
Zm00001d028813 Transducin family protein/WD-40 repeat family protein Chr.1
Zm00001d028814 Pathogenesis-related protein 10 Chr.1
Zm00001d028815 Pathogenesis-related protein 10 Chr.1
Zm00001d028816 Pathogenesis-related protein 10 Chr.1
Zm00001d028818 Calpain-type cysteine protease DEK1 Chr.1

图4

区段内5个预测基因在玉米自交系B73中不同组织的表达量"

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