水稻窄长粒突变体nlg1的筛选及候选基因鉴定

doi:10.16288/j.yczz.24-329

遗传 ›› 2025, Vol. 47 ›› Issue (6): 672-680.doi: 10.16288/j.yczz.24-329

水稻窄长粒突变体nlg1的筛选及候选基因鉴定

郭念¹(), 王辰杰¹, 李钊¹, 韩囡囡¹, 周晨¹, 王凯迎¹, 黄科², 潘咏清²^,³, 李英洁²(), 李云海²()

1.海南大学南繁学院(三亚南繁研究院)，三亚 572025
2.中国科学院遗传与发育生物学研究所，种子创新重点实验室，北京100101
3.中国科学院大学，北京 100039

收稿日期:2024-11-14 修回日期:2024-12-29 出版日期:2025-06-20 发布日期:2025-02-21
通讯作者: 李英洁，博士，博士后，研究方向：水稻籽粒大小调控。E-mail: liyingjie@genetics.ac.cn;
李云海，博士，研究员，研究方向：植物种子和器官大小调控。E-mail: yhli@genetics.ac.cn
作者简介:郭念，硕士研究生，专业方向：作物栽培与耕作。E-mail: 1243323954@qq.com
基金资助:
国家重点研发计划(2021YFF1000202);国家重点研发计划(2022YFF1002903)

Genetic analysis and identification of candidate genes for a narrow and long grain mutant (nlg1) in rice

Nian Guo¹(), Chenjie Wang¹, Zhao Li¹, Nannan Han¹, Chen Zhou¹, Kaiying Wang¹, Ke Huang², Yongqing Pan²^,³, Yingjie Li²(), Yunhai Li²()

1. School of Breeding and Multiplication (Sanya Institute of Breeding and Multiplication), Hainan University, Sanya 572025, China
2. Key Laboratory of Seed Innovation, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China
3. University of Chinese Academy of Sciences, Beijing 100039, China

Received:2024-11-14 Revised:2024-12-29 Published:2025-06-20 Online:2025-02-21
Supported by:
National Key Research and Development Program of China(2021YFF1000202);National Key Research and Development Program of China(2022YFF1002903)

摘要/Abstract

摘要：

籽粒大小是水稻产量的关键决定因素之一，挖掘调控水稻籽粒大小的基因并解析其分子机制，对水稻精准育种具有重要意义。本研究通过EMS诱变粳稻品种ZH11，获得一个籽粒变窄变长的突变体nlg1(narrow and long grain 1)。扫描电镜观察发现nlg1成熟颖壳外表皮纵向细胞数目增加，横向细胞数目减少且细胞宽度变窄，表明NLG1基因可能通过调控颖壳细胞的增殖和扩展影响籽粒大小。结合基因组重测序和MutMap分析，将候选基因定位为LOC_Os09g27590(即已报道的GS9)，该基因编码一个结构域功能未知的蛋白。在nlg1突变体中，GS9基因第一个外显子插入一个碱基C，导致移码突变和蛋白翻译提前终止。遗传互补实验证实NLG1基因能够恢复nlg1突变体的表型。因此，本研究鉴定了一个新的GS9等位突变，为深入解析GS9调控籽粒大小的分子机制提供了新线索，同时为水稻分子设计育种提供了重要的基因资源和理论依据。

关键词: 水稻, 籽粒大小, MutMap, GS9

Abstract:

Grain size is one of the components that determine the rice yield. Exploring more key genes that regulating grain size and analyzing their molecular mechanisms are of great importance for precision breeding of rice. In this study, we screened a mutant nlg1(narrow and long grain 1) with narrow and long grains by mutagenizing the japonica rice variety ZH11 with EMS. Observation and statistical analysis of the outer surface of the mature glume of nlg1 by scanning electron microscopy revealed an increased number of longitudinal cells and a decreased number of transverse cells, along with a reduction in the width of transverse cells. These findings suggest that NLG1 may regulate grain size by simultaneously influencing the proliferation and expansion of lemma cells. The candidate gene was cloned by genome resequencing combined with MutMap analysis method. The results showed that the candidate gene of LOC_Os09g27590 was the previous reported gene, GS9, which encodes a protein with unknown domains. In the nlg1 mutant, a base C was inserted in the first exon of the candidate gene of LOC_Os09g27590, causing a frameshift mutation that leads to premature termination of protein translation. Further genetic complementation experiments showed that NLG1 rescued the phenotype of the nlg1 mutant. Therefore, this study identifies a novel GS9 allele mutation, and further dissection of its molecular regulatory networks in grain size control can provides a theoretical support and genetic resources for precise rice breeding.

Key words: rice, grain size, MutMap, GS9

郭念, 王辰杰, 李钊, 韩囡囡, 周晨, 王凯迎, 黄科, 潘咏清, 李英洁, 李云海. 水稻窄长粒突变体nlg1的筛选及候选基因鉴定[J]. 遗传, 2025, 47(6): 672-680.

Nian Guo, Chenjie Wang, Zhao Li, Nannan Han, Chen Zhou, Kaiying Wang, Ke Huang, Yongqing Pan, Yingjie Li, Yunhai Li. Genetic analysis and identification of candidate genes for a narrow and long grain mutant (nlg1) in rice[J]. Hereditas(Beijing), 2025, 47(6): 672-680.

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引物名称	引物序列(5′→3′)	用途
nlg1seq-F	CTCCTCACTCATCCTACTC	鉴定nlg1突变位点引物
nlg1seq-R	AAGAAACTGTTGCCTTTGCTCT	鉴定nlg1突变位点引物
COM-F	TGACCATGATTACGAATTCGAGCTCCTTAAGCTCACTGACCTCAC	构建互补载体引物
COM-R	TAAAACGACGGCCAGTGCCAAGCTTTCCTGCTCTACCCTGTCTTT	构建互补载体引物
COM-JD-F	CTCCTCACTCATCCTACTC	转基因苗鉴定引物
COM-JD-R	TCGTCGTCAGCTAATCATCG	转基因苗鉴定引物

编号	染色体	物理位置	碱基变化	所在位置	基因号	测序频次	突变类型
INDEL1	Chr.9	2826399	A/AT	基因间区	/	0/8	/
SNP1	Chr.9	3547096	A/C	基因间区	/	0/7	/
SNP2	Chr.9	15786577	G/A	5'UTR	LOC_Os05g39250	0/24	/
INDEL2	Chr.9	16691171	G/GC	外显子	LOC_Os09g27590	0/27	移码突变
SNP3	Chr.9	18187015	T/C	基因上游	LOC_Os09g29980	0/15	/
INDEL3	Chr.9	22586699	A/AAC	基因上游	LOC_Os09g39330	0/20	/
INDEL4	Chr.9	23212674	T/TG	基因上游	LOC_Os09g34310	0/5	/

水稻窄长粒突变体nlg1的筛选及候选基因鉴定

Genetic analysis and identification of candidate genes for a narrow and long grain mutant (nlg1) in rice

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