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作物学报 ›› 2025, Vol. 51 ›› Issue (6): 1501-1513.doi: 10.3724/SP.J.1006.2025.43052

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

玉米ZmKL1优异等位基因调控籽粒大小的效应评估及分子机制解析

杨晓慧1,2(), 晏宣军2,3, 杨文妍2, 付俊杰2, 杨琴1,*(), 谢玉心2,*()   

  1. 1西北农林科技大学农学院/作物抗逆与高效生产全国重点实验室/农业农村部西北旱区玉米生物学与遗传育种重点实验室, 陕西杨凌712100
    2中国农业科学院作物科学研究所/作物基因资源与育种全国重点实验室, 北京100081
    3海南大学热带农林学院, 海南海口570228
  • 收稿日期:2024-11-15 接受日期:2025-03-26 出版日期:2025-06-12 网络出版日期:2025-04-01
  • 通讯作者: *谢玉心, E-mail: xieyuxin@caas.cn;杨琴: E-mail: qyang@nwafu.edu.cn
  • 作者简介:E-mail: 507163727@qq.com
  • 基金资助:
    本研究由国家自然科学基金项目(32101809);中国农业科学院科技创新工程项目资助

Effect evaluation and investigation on molecular mechanism of the ZmKL1 favorable allele in regulating maize kernel size

YANG Xiao-Hui1,2(), YAN Xuan-Jun2,3, YANG Wen-Yan2, FU Jun-Jie2, YANG Qin1,*(), XIE Yu-Xin2,*()   

  1. 1College of Agronomy, Northwest A&F University / State Key Laboratory of Crop Stress Resistance and High-Efficiency Production / Key Laboratory of Maize Biology and Genetic Breeding in Arid Area of Northwest Region, Ministry of Agriculture and Rural Affairs, Yangling 712100, Shaanxi, China
    2Institute of Crop Sciences, Chinese Academy of Agricultural Sciences / State Key Laboratory of Crop Gene Resources and Breeding, Beijing 100081, China
    3College of Tropical Agriculture and Forestry, Hainan University, Haikou 570228, Hainan, China
  • Received:2024-11-15 Accepted:2025-03-26 Published:2025-06-12 Published online:2025-04-01
  • Contact: *E-mail: xieyuxin@caas.cn;E-mail: qyang@nwafu.edu.cn
  • Supported by:
    the National Natural Science Foundation of China(32101809);the Innovation Program of Chinese Academy of Agricultural Sciences.

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摘要： 为评估玉米籽粒大小调控基因ZmKL1不同等位基因对农艺性状的效应, 解析其调控籽粒大小的分子机制, 本研究构建了近等基因系(NIL), 在2个地点调查其田间表型、穗部及籽粒表型, 并进行转录组与蛋白组测序, 共同解析不同等位基因对籽粒大小的调控效应。结果显示, NIL群体在粒长、粒宽、百粒重、株高及穗位高方面存在显著差异, 花期及穗部表型等方面无差异。2组NIL间差异表达基因总数为744个、差异表达蛋白总数为152个。GO分析发现差异表达基因富集于蛋白结合、氧化还原酶活性等途径, 差异表达蛋白则与转录、基因表达调控、RNA生物合成及代谢调控过程密切相关。进一步利用荧光定量PCR试验对8个关键基因的表达差异进行了验证。本研究对ZmKL1等位基因进行了表型评估, 发现ZmKL1优异等位基因在玉米产量性状改良方面具有一定的应用潜力; 同时联合转录组与蛋白组初步解析了其调控籽粒大小的机制, 为进一步挖掘影响籽粒大小关键基因及通路提供了思路。

关键词: 玉米籽粒, 转录组, 蛋白组, 等位基因, 近等基因系

Abstract:

To evaluate the effects of different alleles of the kernel size-related gene ZmKL1 on agronomic traits and to elucidate the molecular mechanisms by which ZmKL1 regulates kernel size in maize, this study constructed near-isogenic lines (NILs) and analyzed their field performance, ear morphology, and kernel traits at two locations. Transcriptome and proteome analyses were conducted to explore the regulatory effects of different alleles on kernel size. The results revealed significant differences in kernel length, kernel width, hundred-kernel weight, plant height, and ear height among the NILs, while no significant differences were observed in flowering time or ear traits. A total of 744 differentially expressed genes (DEGs) and 152 differentially expressed proteins (DEPs) were identified between the two NIL groups. Gene Ontology (GO) analysis indicated that DEGs were enriched in pathways related to protein binding and oxidoreductase activity, while DEPs were primarily associated with transcriptional regulation, gene expression, RNA biosynthesis, and metabolic processes. The expression differences of eight key genes were further validated by quantitative real-time PCR (qRT-PCR). This study not only provides a comprehensive phenotypic assessment of ZmKL1 alleles, demonstrating the potential of the favorable allele for maize yield improvement, but also offers preliminary insights into the molecular mechanisms underlying ZmKL1-mediated kernel size regulation through integrative transcriptomic and proteomic analyses. These findings contribute to the identification of key genes and pathways involved in maize kernel development, laying the foundation for future genetic improvement strategies.

Key words: maize kernel, transcriptome, proteome, allele, near-isogenic line

图1

近等基因系群体构建 A: 等位基因测序图; B: 近等基因系构建流程图; C: 近等基因系ZmKL1基因序列比对; D: 近等基因系ZmKL1蛋白序列比对。NIL1代表携带插入变异(CTCAG)的近等基因系; NIL2代表携带缺失变异(-----)的近等基因系。"

表1

试验所用引物"

引物名称
Primer name		 引物序列
Primer sequence (5'-3')		 引物用途
Primer usage
KL1-genomic-F1 GGTTGTACCACACCTGCCA 分段PCR Segmented PCR
KL1-genomic-R1 GGGGTACACCCAAAGGCT 分段PCR Segmented PCR
KL1-genomic-F2 TAGCTGTGCGTTCAAGTGGT 分段PCR Segmented PCR
KL1-genomic-R2 CGCCGCCAGGATTTGTTG 分段PCR Segmented PCR
Zm00001d000023-F GGCAAATGTCTGATGTGCAGGAGG 实时荧光定量PCR qRT-PCR
Zm00001d000023-R GCCCTTATTGCTCCAGCAAGAGG 实时荧光定量PCR qRT-PCR
Zm00001d010525-F CCTGGTGCAACACCACTAGAGAG 实时荧光定量PCR qRT-PCR
Zm00001d010525-R TTCTGCTCTCCTCGCGCCAAT 实时荧光定量PCR qRT-PCR
Zm00001d010640-F CAAGGCACGAATCCACGCCGA 实时荧光定量PCR qRT-PCR
Zm00001d010640-R GAGGGAAACGAGCCCACAGAGC 实时荧光定量PCR qRT-PCR
Zm00001d013193-F CTTCAGCTCCAGCCGCAGCAT 实时荧光定量PCR qRT-PCR
Zm00001d013193-R CTGCAGCACTCATCGCCTTCG 实时荧光定量PCR qRT-PCR
Zm00001d034160-F GAGCCTCGAGGAGGAGCGCC 实时荧光定量PCR qRT-PCR
Zm00001d034160-R GTCGTCGGCCTTGGCCTCCG 实时荧光定量PCR qRT-PCR
Zm00001d044104-F TGTTCCACACCTCCGCCTTCGT 实时荧光定量PCR qRT-PCR
Zm00001d044104-R CCTCCACACACATGCCAGCAAG 实时荧光定量PCR qRT-PCR
Zm00001d044156-F TGTAGCCATGACGTCGTTCCCC 实时荧光定量PCR qRT-PCR
Zm00001d044156-R CGTGAACTTCTCGAAGTGGCCAAAC 实时荧光定量PCR qRT-PCR
Zm00001d051938-F TGCACATGGACCCCAGCCGGA 实时荧光定量PCR qRT-PCR
Zm00001d051938-R CGCCGTCACCTCCGCCATCAT 实时荧光定量PCR qRT-PCR
ZmActin-F ATGGTCAAGGCCGGTTTCG 实时荧光定量PCR qRT-PCR
ZmActin-R TCAGGATGCCTCTCTTGGCC 实时荧光定量PCR qRT-PCR

图2

自然材料里不同单倍型的籽粒表型鉴定 A~C分别为粒长、粒宽、百粒重的统计数据分析结果。*: P < 0.05; ***: P < 0.001, Student’s t测验。Hap1代表携带插入变异(CTCAG)的单倍型; Hap2代表携带缺失变异(-----)的单倍型。"

图3

近等基因系的田间表型鉴定图(A~E)、穗部表型鉴定图(F~I)、穗部表型图(J) BJ: 北京; LF: 廊坊。*: P < 0.05, ****: P < 0.0001, ns: 无显著性。Student’s t测验。NIL1为携带插入变异(CTCAG)的近等基因系; NIL2为携带缺失变异(-----)的近等基因系。标尺为1 cm。"

图4

近等基因系的籽粒表型鉴定 A~C分别为粒长、粒宽、百粒重测量图, D~F分别为粒长、粒宽、百粒重的统计数据分析结果。BJ: 北京; LF: 廊坊。***: P < 0.001;****: P < 0.0001, Student’s t测验。标尺为1 cm。NIL1代表携带插入变异(CTCAG)的近等基因系; NIL2代表携带缺失变异(-----)的近等基因系。"

图5

NIL1和NIL2授粉后10 d (A, B)与15 d (C, D)的石蜡切片图 标尺为1 mm。"

图6

NIL群体转录组分析 A: NIL1和NIL2差异表达基因火山图; B: 差异表达基因GO分析图。"

图7

NIL群体蛋白组分析 A: 肽段长度分布图; B: 蛋白分子量与等电点分布图; C: NIL1和NIL2差异表达蛋白火山图; D: 差异蛋白的BP富集气泡图(排名前20)。"

图8

转录组和蛋白组联合分析 A: 差异表达基因与差异表达蛋白韦恩图; B: 差异表达基因与差异表达蛋白相关性图。DEGs: 差异表达基因; DAPs: 差异表达蛋白。"

图9

差异表达基因的qPCR验证分析 Student’s t检验, **: P < 0.01; ***: P < 0.001; ****: P < 0.0001。"

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