水稻粉质胚乳突变体we1的表型分析与基因定位

doi:10.16819/j.1001-7216.2025.240508

中国水稻科学 ›› 2025, Vol. 39 ›› Issue (4): 543-551.DOI: 10.16819/j.1001-7216.2025.240508

水稻粉质胚乳突变体we1的表型分析与基因定位

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

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，南京 210014
²生物育种钟山实验室，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:2024-05-15 修回日期:2024-08-20 出版日期:2025-07-10 发布日期:2025-07-21
通讯作者: ^*email: yangjie168@aliyun.com
基金资助:
国家自然科学基金资助项目(32201861);江苏省重点研发计划资助项目(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 we1 in Rice

ZHU Jianping¹^,²^,³, LI Xia¹^,²^,³, LI Wenqi¹^,²^,³, XU Yang¹^,²^,³, WANG Fangquan¹^,²^,³, TAO Yajun¹^,²^,³, JIANG Yanjie¹^,²^,³, CHEN Zhihui¹^,²^,³, FAN Fangjun¹^,²^,³, YANG Jie¹^,²^,³^,^*()

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, China
²Zhongshan Biological Breeding Laboratory, Nanjing 210014, China
³Jiangsu Co-innovation Center for Modern Production Technology of Grain Crops, Yangzhou University, Yangzhou 225009, China

Received:2024-05-15 Revised:2024-08-20 Online:2025-07-10 Published:2025-07-21
Contact: ^*email: yangjie168@aliyun.com

摘要/Abstract

摘要：

【目的】对粉质胚乳突变体的研究有助于解析稻米品质形成的分子机制。【方法】从日本晴⁶⁰Co诱变突变体库中筛选到一份粉质皱缩胚乳突变体we1(white endosperm1)，对其表型、理化性质进行分析，并利用we1与9311杂交获得的F₂群体对目标基因进行精细定位。【结果】we1胚乳表现为白色粉质皱缩状，淀粉颗粒排列疏松且不规则；千粒重、株高、总淀粉、直链淀粉含量显著下降，脂肪含量显著上升。遗传分析表明we1粉质皱缩胚乳性状受单个隐性基因控制。利用we1/9311 F₂群体进行基因定位，WE1被定位在6号染色体短臂P16和P18之间约181 kb区间内，该区间包含23个开放阅读框(Open reading frame, ORFs)。qRT-PCR 结果显示，WE1的突变会影响淀粉合成相关基因的表达。【结论】WE1可能是一个参与淀粉合成的新基因，本研究为进一步解析WE1调控水稻淀粉合成的分子机制奠定了基础。

关键词: 水稻, 粉质胚乳, 淀粉, 理化性质, 精细定位

Abstract:

【Objective】 The study of floury endosperm mutants is helpful for elucidating the molecular mechanisms underlying rice grain quality formation. 【Method】 A stably inherited floury endosperm mutant we1 (white endosperm 1) was isolated from a ⁶⁰Co-irradiated mutant pool of japonica rice cultivar Nipponbare. Phenotypic characterization and physicochemical analyses of we1 were conducted. An F₂ population derived from a cross between we1 and indica variety 9311 was used for fine mapping. 【Result】 Compared with the wild type, we1 exhibited white, shrunken endosperm with irregularly shaped and loosely packed compound starch granules. The mutant showed significant reductions in 1000-grain weight, plant height, total starch content, and amylose content, along with a marked increase in lipid content. Genetic analysis indicated that the mutant phenotype was controlled by a single recessive nuclear gene. For map-based cloning, we1 was crossed with 9311 and recessive F₂ individuals were selected. The WE1 locus was initially mapped to chromosome 6 and fine-mapped to a 181-kb physical interval. Twenty-three open reading frames (ORFs) were predicted in this region. qRT-PCR analysis revealed altered expression levels of starch synthesis-related genes in the we1 mutant. 【Conclusion】 WE1 likely encodes a novel gene involved in starch synthesis, providing a foundation for further elucidating its molecular regulatory mechanisms in rice.

Key words: rice, floury endosperm, starch, physicochemical property, fine mapping

朱建平, 李霞, 李文奇, 许扬, 王芳权, 陶亚军, 蒋彦婕, 陈智慧, 范方军, 杨杰. 水稻粉质胚乳突变体we1的表型分析与基因定位[J]. 中国水稻科学, 2025, 39(4): 543-551.

ZHU Jianping, LI Xia, LI Wenqi, XU Yang, WANG Fangquan, TAO Yajun, JIANG Yanjie, CHEN Zhihui, FAN Fangjun, YANG Jie. Phenotypic Analysis and Gene Mapping of a Floury Endosperm Mutant we1 in Rice[J]. Chinese Journal OF Rice Science, 2025, 39(4): 543-551.

图/表 11

参考文献 31

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引物名称 Primer name	正向引物序列 Forward primer (5’-3’)	反向引物序列 Reverse primer(5’-3’)
P1	CCCCAACTTTCAGCTTTGCT	GACAAAATCCTCCGTGGCG
P5	AATGCGAAGTTTGGGGGAAC	CAATCAGACGACGAAACGGG
P8	CGTGGACGGTGATGGAATTC	CCTCAGTGACATGCTCAAGG
P12	ACTCAGTAGCAGTTGAGGAGA	TCAAATCCTTGCACATAACCCA
P16	ACTCAGCCGATACTTGAGGG	TTTGGAGTCTATTAGGCTTGAAGC
P18	TGCATGTGTTCACACTGTGA	ACTAATGATCAGCTAGGTGCCT

引物名称 Primer name	正向引物序列 Forward primer (5’-3’)	反向引物序列 Reverse primer(5’-3’)
P1	CCCCAACTTTCAGCTTTGCT	GACAAAATCCTCCGTGGCG
P5	AATGCGAAGTTTGGGGGAAC	CAATCAGACGACGAAACGGG
P8	CGTGGACGGTGATGGAATTC	CCTCAGTGACATGCTCAAGG
P12	ACTCAGTAGCAGTTGAGGAGA	TCAAATCCTTGCACATAACCCA
P16	ACTCAGCCGATACTTGAGGG	TTTGGAGTCTATTAGGCTTGAAGC
P18	TGCATGTGTTCACACTGTGA	ACTAATGATCAGCTAGGTGCCT

引物名称	正向引物序列	反向引物序列
Primer name	Forward primer(5’-3’)	Reverse primer(5’-3’)
Actin	CATGCTATCCCTCGTCTCGACCT	CGCACTTCATGATGGAGTTGTAT
AGPS1	AGAATGCTCGTATTGGAGAAAATG	GGCAGCATGGAATAAACCAC
AGPS2b	AACAATCGAAGCGCGAGAAA	GCCTGTAGTTGGCACCCAGA
AGPL1	GGAAGACGGATGATCGAGAAAG	CACATGAGATGCACCAACGA
AGPL2	AGTTCGATTCAAGACGGATAGC	CGACTTCCACAGGCAGCTTATT
AGPL4	TCAACGTCGATGCAGCAAAT	ATCCCTCAGTTCCTAGCCTCATT
GBSSI	TCCGAGAGGTTCAGGTCATC	ATGAGCTCCTCGGCGTAGTA
SSI	GGGCCTTCATGGATCAACC	CCGCTTCAAGCATCCTCATC
SSIIa	GCTTCCGGTTTGTGTGTTCA	CTTAATACTCCCTCAACTCCACCAT
SSIIc	GCAAAGCGGCTATAGAGGTTCC	ATTTCCATCACCGTAGCAGTAGC
SSIIIa	GCCTGCCCTGGACTACATTG	GCAAACATATGTACACGGTTCTGG
SSIVa	GGGAGCGGCTCAAACATAAA	CCGTGCACTGACTGCAAAAT
BEI	TGGCCATGGAAGAGTTGGC	CAGAAGCAACTGCTCCACC
BEIIa	AGCGCTGAAGGCATTACCTACC	CTACTAATGCTGCAGACTGTGCTC
BEIIb	ATGCTAGAGTTTGACCGC	AGTGTGATGGATCCTGCC
ISA1	TGCTCAGCTACTCCTCCATCATC	AGGACCGCACAACTTCAACATA
ISA2	TAGAGGTCCTCTTGGAGG	AATCAGCTTCTGAGTCACCG
ISA3	ACAGCTTGAGACACTGGGTTGAG	GCATCAAGAGGACAACCATCTG

引物名称	正向引物序列	反向引物序列
Primer name	Forward primer(5’-3’)	Reverse primer(5’-3’)
Actin	CATGCTATCCCTCGTCTCGACCT	CGCACTTCATGATGGAGTTGTAT
AGPS1	AGAATGCTCGTATTGGAGAAAATG	GGCAGCATGGAATAAACCAC
AGPS2b	AACAATCGAAGCGCGAGAAA	GCCTGTAGTTGGCACCCAGA
AGPL1	GGAAGACGGATGATCGAGAAAG	CACATGAGATGCACCAACGA
AGPL2	AGTTCGATTCAAGACGGATAGC	CGACTTCCACAGGCAGCTTATT
AGPL4	TCAACGTCGATGCAGCAAAT	ATCCCTCAGTTCCTAGCCTCATT
GBSSI	TCCGAGAGGTTCAGGTCATC	ATGAGCTCCTCGGCGTAGTA
SSI	GGGCCTTCATGGATCAACC	CCGCTTCAAGCATCCTCATC
SSIIa	GCTTCCGGTTTGTGTGTTCA	CTTAATACTCCCTCAACTCCACCAT
SSIIc	GCAAAGCGGCTATAGAGGTTCC	ATTTCCATCACCGTAGCAGTAGC
SSIIIa	GCCTGCCCTGGACTACATTG	GCAAACATATGTACACGGTTCTGG
SSIVa	GGGAGCGGCTCAAACATAAA	CCGTGCACTGACTGCAAAAT
BEI	TGGCCATGGAAGAGTTGGC	CAGAAGCAACTGCTCCACC
BEIIa	AGCGCTGAAGGCATTACCTACC	CTACTAATGCTGCAGACTGTGCTC
BEIIb	ATGCTAGAGTTTGACCGC	AGTGTGATGGATCCTGCC
ISA1	TGCTCAGCTACTCCTCCATCATC	AGGACCGCACAACTTCAACATA
ISA2	TAGAGGTCCTCTTGGAGG	AATCAGCTTCTGAGTCACCG
ISA3	ACAGCTTGAGACACTGGGTTGAG	GCATCAAGAGGACAACCATCTG

杂交组合 Cross	正常透明种子数 No. of wild type seeds	突变粉质种子数 No. of mutant seeds	χ²_（3:1）
we1/野生型F₂ F₂of we1/wild type	132	50	0.469
野生型/we1 F₂ F₂ of wild type/we1	151	47	0.108

水稻粉质胚乳突变体we1的表型分析与基因定位

Phenotypic Analysis and Gene Mapping of a Floury Endosperm Mutant we1 in Rice

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开放阅读框	基因登录号	功能注释
ORFs	Locus ID	Functional description
ORF1	Os06g0165200	含有仙茅凝集素结构域的蛋白质Curculin-like lectin domain containing protein
ORF2	Os06g0165300	保守假定蛋白Conserved hypothetical protein
ORF3	Os06g0165500	含有仙茅凝集素结构域的蛋白质Curculin-like lectin domain containing protein
ORF4	Os06g0165600	含有AP2结构域的蛋白质AP2 domain containing protein
ORF5	Os06g0165800	咖啡酰辅酶A氧甲基转移酶Caffeoyl-CoA 3-O-methyltransferase
ORF6	Os06g0166000	含有F-box结构域的蛋白质Cyclin-like F-box domain containing protein
ORF7	Os06g0166100	含有FAR1结构域的蛋白质FAR1 domain containing protein
ORF8	Os06g0166200	含有C2H2型锌指结构域的蛋白质Zinc finger, C2H2-type domain containing protein
ORF9	Os06g0166400	TINY 类蛋白质TINY-like protein
ORF10	Os06g0166500	生长素响应蛋白IAA20 Auxin-responsive protein IAA20
ORF11	Os06g0166900	含有蛋白激酶结构域的蛋白质Protein kinase-like domain containing protein
ORF12	Os06g0167000	PRP8 蛋白质PRP8 protein
ORF13	Os06g0167100	含有Armadillo类螺旋结构域的蛋白质Armadillo-like helical domain containing protein
ORF14	Os06g0167200	RING-H2型锌指蛋白RING-H2 finger protein ATL1R
ORF15	Os06g0167400	DTC家族蛋白Di-trans-poly-cis-decaprenylcistransferase family protein
ORF16	Os06g0167500	含有LRR结构域的蛋白质Leucine-rich repeat, plant specific containing protein
ORF17	Os06g0167600	蛋白酶体亚基-3 Proteasome subunit alpha-3
ORF18	Os06g0168000	谷胱甘肽硫转移酶Glutathione S-transferase, C-terminal-like domain containing protein
ORF19	Os06g0168400	保守假定蛋白Conserved hypothetical protein
ORF20	Os06g0168500	Syntaxin类蛋白 Syntaxin-like protein
ORF21	Os06g0168600	核糖核苷酸还原酶Ribonucleotide reductase
ORF22	Os06g0168700	富含脯氨酸蛋白Prolin rich protein
ORF23	Os06g0168800	蛋白激酶Protein kinase

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