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作物学报

作物学报 ›› 2025, Vol. 51 ›› Issue (4): 1110-1117.doi: 10.3724/SP.J.1006.2025.42042

• 研究简报 • 上一篇    下一篇

水稻粉质胚乳突变体we2的表型分析与基因定位

朱建平1,2,3(), 李文奇1,2,3, 许扬1,2,3, 王芳权1,2,3, 李霞1,2,3, 蒋彦婕1,2,3, 范方军1,2,3, 陶亚军1,2,3, 陈智慧1,2,3, 吴莹莹1,2,3, 杨杰1,2,3,*()   

  1. 1江苏省农业科学院粮食作物研究所 / 农业农村部淮河下游种质创制重点实验室(南京) / 国家水稻改良中心南京分中心, 江苏南京 210014
    2生物育种钟山实验室, 江苏南京 210014
    3扬州大学江苏省粮食作物现代产业技术协同创新中心, 江苏扬州 225009
  • 收稿日期:2024-09-07 接受日期:2024-12-12 出版日期:2025-04-12 网络出版日期:2024-12-27
  • 通讯作者: 杨杰, E-mail: yangjie168@aliyun.com
  • 作者简介:E-mail: 1525209729@qq.com
  • 基金资助:
    国家自然科学基金项目(32201861);江苏省重点研发计划项目(BE2023362);江苏省种业振兴揭榜挂帅项目(JBGS (2021) 008)

Phenotypic analysis and gene mapping of a floury endosperm mutant we2 in rice

ZHU Jian-Ping1,2,3(), LI Wen-Qi1,2,3, XU Yang1,2,3, WANG Fang-Quan1,2,3, LI Xia1,2,3, JIANG Yan-Jie1,2,3, FAN Fang-Jun1,2,3, TAO Ya-Jun1,2,3, CHEN Zhi-Hui1,2,3, WU Ying-Ying1,2,3, YANG Jie1,2,3,*()   

  1. 1Institute of Food Crops, Jiangsu Academy of Agricultural Sciences / Key Laboratory of Germplasm Innovation in Downstream of Huaihe River (Nanjing), Ministry of Agricultural and Rural Affairs / Nanjing Branch of Chinese National Center for Rice Improvement, Nanjing 210014, Jiangsu, China
    2Zhongshan Biological Breeding Laboratory, Nanjing 210014, Jiangsu, China
    3Jiangsu Co-Innovation Center for Modern Production Technology of Grain Crops, Yangzhou University, Yangzhou 225009, Jiangsu, China
  • Received:2024-09-07 Accepted:2024-12-12 Published:2025-04-12 Published online:2024-12-27
  • Contact: E-mail: yangjie168@aliyun.com
  • Supported by:
    National Natural Science Foundation of China(32201861);Jiangsu Provincial Key Research and Development Program(BE2023362);Project of Seeds Innovation in Jiangsu(JBGS (2021) 008)

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摘要：

从日本晴60Co诱变突变体库中筛选到一份粉质胚乳突变体we2 (white endosperm2), 对其表型和理化性质进行分析, 并利用we2与9311杂交获得的F2群体对目标基因进行精细定位。we2胚乳表现为白色粉质, 淀粉颗粒排列疏松且不规则; 千粒重、总淀粉和直链淀粉含量显著下降, 脂肪含量显著上升。遗传分析表明we2粉质胚乳性状受单个隐性基因控制。利用we2/9311 F2群体进行基因定位, WE2被定位在第6染色体长臂P38和P39之间约244 kb区间内, 该区间包含27个开放阅读框(Open reading frames, ORFs)。qRT-PCR结果显示, we2胚乳中淀粉合成相关基因表达显著下调。本研究为WE2的克隆和功能分析奠定了基础。

关键词: 水稻, 胚乳, 淀粉, 精细定位

Abstract:

A floury endosperm mutant, we2 (white endosperm2), was identified from a 60Co-irradiated mutant pool of the rice (Oryza sativa) variety Nipponbare. The phenotype and physicochemical properties of we2 were analyzed, and an F2 population derived from a cross between we2 and the indica variety 9311 was used for fine mapping of the target gene. Compared with the wild type, we2 exhibited a white endosperm phenotype with irregularly shaped, loosely packed compound starch granules. The 1000-grain weight, total starch content, and amylose content in we2 were significantly lower than those in the wild type, whereas the lipid content was higher. Genetic analysis revealed that the floury endosperm phenotype of we2 is controlled by a single nuclear recessive gene. For map-based cloning, the we2 mutant was crossed with 9311, and F2 individuals were analyzed. The WE2 locus was initially mapped to chromosome 6 and subsequently fine-mapped to a 244 kb genomic region containing 27 predicted open reading frames (ORFs). Quantitative real-time PCR (qRT-PCR) analysis demonstrated that the expression levels of several genes involved in starch biosynthesis were reduced in the we2 mutant. This study provides a foundation for the cloning and functional characterization of WE2, contributing to a deeper understanding of the genetic and molecular mechanisms underlying rice endosperm development.

Key words: rice, endosperm, starch, fine mapping

表1

WE2精细定位引物"

名称
Name		 正向引物
Forward primer (5′-3′)		 反向引物
Reverse primer (5′-3′)
P22 GACGACACGCACAGCATC GGGGGTTTATATTGATTTTTGCGA
P27 GGACATACCTAGCGCTAGC AGTACTCTGATCACCGTCAGT
P31 CGAGGACGACAGAACGAAGA CGTGTTGAGCCCCACCAG
P36 TGTTTGGAGAGAGTTGGAGATT GGATCGGTTTTCACAAACAGAC
P38 AACAATTCACTCGCCACAGC CGGCGGTTTTGGATTTGGAT
P39 AGCACAGTACTATCGACGGG GGCCATTTTGTGCTGGTACA

图1

表型分析 A, B: WT (A)与we2 (B)种子; C, D: WT (C)与we2 (D)种子横切面; E: 粒长, n = 20; F: 粒宽, n = 20; G: 粒厚, n = 20; 标尺为1 mm。数值为平均值±标准差, 采用t测验, **表示P < 0.01。"

图2

农艺性状分析 A~D: 千粒重、株高、分蘖数和结实率, n = 20。数值为平均值±标准差, 采用t测验, **表示P < 0.01。"

图3

理化性质分析 A~D: 总淀粉含量、直链淀粉含量、脂肪含量和总蛋白含量, n = 3。数值为平均值±标准差, 采用t测验, **表示P < 0.01。"

图4

扫描电镜分析 WT (A~C)和we2 (D~F)种子横切面观察。A, D标尺为1 mm; B, E标尺为50 μm; C, F标尺为10 μm。"

表2

遗传分析"

杂交组合
Cross combination		 野生型种子数
No. of wild type seeds		 突变体种子数
No. of mutant seeds		 χ2(3:1)
we2/wild type F2 127 40 0.050
Wild type/we2 F2 148 55 0.369

图5

WE2的精细定位 WE2定位在6号染色体长臂端标记P38和P39区间内, 物理距离约为244 kb, 包括27个候选基因。"

表3

WE2候选基因"

开放阅读框
ORFs		 基因登录号
Locus ID		 功能注释
Functional description
ORF1 Os06g0659400 Protein of unknown function DUF231, plant domain containing protein
ORF2 Os06g0659500 Similar to Glutaredoxin
ORF3 Os06g0659800 Conserved hypothetical protein
ORF4 Os06g0660200 Similar to Auxin efflux carrier protein
ORF5 Os06g0660400 Conserved hypothetical protein
ORF6 Os06g0660600 Homeodomain-like containing protein
ORF7 Os06g0660700 Similar to Ubiquitin-conjugating enzyme E2S
ORF8 Os06g0660800 Protein kinase domain containing protein
ORF9 Os06g0661000 Plant MuDR transposase domain containing protein
ORF10 Os06g0661400 ANTH domain containing protein
ORF11 Os06g0661500 Conserved hypothetical protein
ORF12 Os06g0661600 Zinc finger, DHP-type domain containing protein
ORF13 Os06g0661700 RabGAP/TBC domain containing protein
ORF14 Os06g0661800 Similar to Cryptochrome dash
ORF15 Os06g0661900 Protein of unknown function DUF266, plant family protein
ORF16 Os06g0662000 Conserved hypothetical protein
ORF17 Os06g0662200 Eukaryotic transcription factor, DNA-binding domain containing protein
ORF18 Os06g0662300 Pollen allergen Lol p2 family protein
ORF19 Os06g0662500 Pollen allergen Lol p2 family protein
ORF20 Os06g0662600 Pollen allergen Lol p2 family protein
ORF21 Os06g0662700 Pollen allergen Lol p2 family protein
ORF22 Os06g0662800 Pollen allergen Lol p2 family protein
ORF23 Os06g0662900 Pollen allergen/expansion, C-terminal domain containing protein
ORF24 Os06g0663100 Hypothetical protein
ORF25 Os06g0663200 Similar to Protein kinase APK1B, chloroplast precursor
ORF26 Os06g0663300 Pollen allergen Lol p2 family protein
ORF27 Os06g0663400 Serine/threonine protein kinase-like protein

图6

WE2候选基因表达 开花后12 d胚乳中WE2候选基因表达检测(qRT-PCR), 以Actin作为内参, n = 3。数值为平均值±标准差, 采用t测验, **表示P < 0.01。qRT-PCR assay of the expression of WE2 candidate genes in 12 DAF endosperm. Actin was used as control. n = 3. Values are means ± SD, using Student’s t-test. ** indicates P < 0.01."

图7

淀粉合成相关基因表达 开花后12 d胚乳中WE2候选基因表达检测(qRT-PCR), 以Actin作为内参, n = 3. 数值为平均值±标准差, 采用t测验, **表示P < 0.01。"

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