绿豆籽粒蛋白质与苏氨酸含量全基因组关联分析及KASP分子标记开发

doi:10.3864/j.issn.0578-1752.2026.13.003

中国农业科学 ›› 2026, Vol. 59 ›› Issue (13): 2789-2801.doi: 10.3864/j.issn.0578-1752.2026.13.003

• 作物遗传育种·种质资源·分子遗传学 • 上一篇下一篇

绿豆籽粒蛋白质与苏氨酸含量全基因组关联分析及KASP分子标记开发

陈圣琦¹(), 刘金洋²(), 黄佳丽³, 刘肖岑³, 赵硕谦⁴, 林云², 薛晨晨², 闫强², 陈景斌², 吴然然², 陈新²^,³(), 程金平¹(), 袁星星²^,³()

¹ 南京农业大学农学院，南京 210095
² 江苏农业科学院经济作物研究所/江苏省园艺作物遗传改良与高效利用重点实验室，南京 210014
³ 南京农业大学生命科学学院，南京 210095
⁴ 南京农业大学植物保护学院，南京 210095

收稿日期:2025-11-19 接受日期:2026-01-13 出版日期:2026-07-01 发布日期:2026-07-01
通信作者:
陈新，E-mail：cx@jaas.ac.cn
程金平，E-mail：cjp@njau.edu.cn
袁星星，E-mail：yxx@jaas.ac.cn
联系方式: 陈圣琦，E-mail：2023801189@stu.njau.edu.cn。刘金洋，E-mail：20200024@jaas.ac.cn。陈圣琦和刘金洋为同等贡献作者。
基金资助:
国家自然科学基金(32200499); 国家重点研发计划(2023YFD1202702-3); 国家食用豆产业技术体系生物防治与综合防控岗位科学家(CARS-08-G15); 江苏省种业揭榜挂帅项目(JBGS〔2021〕004)

Whole Genome Association Study of Protein and Threonine Content in Mung Bean Seeds and Development of KASP Molecular Marker

CHEN ShengQi¹(), LIU JinYang²(), HUANG JiaLi³, LIU XiaoCen³, ZHAO ShuoQian⁴, LIN Yun², XUE ChenChen², YAN Qiang², CHEN JingBin², WU RanRan², CHEN Xin²^,³(), CHENG JinPing¹(), YUAN XingXing²^,³()

¹ College of Agriculture, Nanjing Agricultural University, Nanjing 210095
² Institute of Economic Crops, Jiangsu Academy of Agricultural Sciences/Key Laboratory of Genetic Improvement of Efficient Horticultural Crops, Nanjing 210014
³ College of Life Sciences, Nanjing Agricultural University, Nanjing 210095
⁴ College of Plant Protection, Nanjing Agricultural University, Nanjing 210095

Received:2025-11-19 Accepted:2026-01-13 Published:2026-07-01 Online:2026-07-01

摘要/Abstract

摘要：

【目的】蛋白质与苏氨酸是绿豆籽粒的核心营养成分，对绿豆营养品质具有关键影响。绿豆蛋白因易消化吸收、生物活性丰富，是素食者的优质蛋白来源，对人体健康极具价值。挖掘与绿豆籽粒蛋白质含量和苏氨酸相对含量性状显著关联的遗传位点及候选基因，为绿豆籽粒蛋白质和苏氨酸含量性状的遗传改良提供理论依据。【方法】利用凯氏定氮法和液相色谱-质谱联用技术，对2022年种植的279份绿豆品种籽粒的蛋白质含量和苏氨酸相对含量进行测定，利用Tassel5软件的一般线性模型进行全基因组关联分析，挖掘绿豆籽粒蛋白质含量和苏氨酸相对含量相关的遗传位点。结合连锁不平衡分析、转录组比较分析和比较基因组学鉴定到与氮代谢相关的关键候选基因。【结果】鉴定出位于第3染色体939 296—1 039 749 bp区域内的6个关键位点，可解释6.06%—12.20%的表型变异。这6个位点之间紧密连锁，主要可以被分成2种单倍型，其中，Hap2为优势单倍型。然后对关键候选位点SLG03_966208（A/G）设计KASP分子标记，可将80份绿豆品种成功分成AA、GG和AG 3种基因型，准确率达87.5%。在位点连锁区域内鉴定到关键候选基因EVM0000757，其拟南芥同源基因AT5G65750与氮代谢显著相关，在高低蛋白品种中，EVM0000757表达差异显著（P=1.84E-03，|log₂FC|=1.39），是可靠候选基因。【结论】鉴定了279份绿豆品种的籽粒蛋白质含量和苏氨酸相对含量。检测到位于第3染色体939 296—1 039 749 bp区域内的6个SNP位点，开发了SLG03_966208（A/G）位点的KASP分子标记，可以区分绿豆籽粒蛋白质和苏氨酸含量的高低。鉴定到与氮代谢相关的关键候选基因EVM0000757。

关键词: 绿豆, 籽粒蛋白质含量, 籽粒苏氨酸相对含量, 全基因组关联分析, KASP标记

Abstract:

【Objective】Protein and threonine are the core nutrients of mung bean seeds, which have a critical impact on the nutritional quality of mung beans. Among them, mung bean protein is a high-quality protein source for vegetarians due to its easy digestion, absorption, and rich biological activity, which is of great value to human health. This study aims to explore genetic loci and candidate genes significantly associated with the protein content and relative content of threonine in mung bean seeds, providing a theoretical basis for genetic improvement of protein and threonine content in mung bean seeds. 【Method】This study used Kjeldahl nitrogen determination method and liquid chromatography-mass spectrometry to determine the protein content and relative content of threonine in 279 mung bean varieties planted in 2022. The general linear model of Tassel5 software was used for genome-wide association study to explore genetic loci related to protein content and relative content of threonine in mung bean seeds. By combining linkage disequilibrium analysis, transcriptome comparative analysis, and comparative genomics, key candidate genes related to nitrogen metabolism were identified. 【Result】Six key loci located on chromosome 3 were identified, which are related to the protein content and relative content of threonine in mung bean seeds, and can explain 6.06% to 12.20% of phenotypic variation. These 6 loci are closely linked and can be mainly divided into two haplotypes, with Hap2 being the dominant haplotype. Then, KASP molecular marker was designed for the key candidate locus SLG03_966208 (A/G), which can successfully classify 80 mung bean varieties into three genotypes: AA, GG, and AG, with a detection accuracy rate of 87.5%. The key candidate gene EVM0000757 was identified within the locus linkage region, and the homologous gene AT5G65750 in Arabidopsis was significantly associated with nitrogen metabolism. Its gene expression was significantly different between high and low protein varieties (P=1.84E-03, |log₂FC|=1.39), making it a reliable candidate gene.【Conclusion】279 mung bean varieties were identified for their grain protein content and relative content of threonine. Six SNP loci were detected within the 939 296-1 039 749 bp region of chromosome 3. A KASP molecular marker for the SLG03_966208 (A/G) locus was developed, which can distinguish the high and low levels of protein and threonine content in mung bean grains. A key candidate gene related to nitrogen metabolism, EVM0000757, was identified.

Key words: mung bean, grain protein content, relative content of grain threonine content, genome-wide association study, KASP marker

陈圣琦, 刘金洋, 黄佳丽, 刘肖岑, 赵硕谦, 林云, 薛晨晨, 闫强, 陈景斌, 吴然然, 陈新, 程金平, 袁星星. 绿豆籽粒蛋白质与苏氨酸含量全基因组关联分析及KASP分子标记开发[J]. 中国农业科学, 2026, 59(13): 2789-2801.

CHEN ShengQi, LIU JinYang, HUANG JiaLi, LIU XiaoCen, ZHAO ShuoQian, LIN Yun, XUE ChenChen, YAN Qiang, CHEN JingBin, WU RanRan, CHEN Xin, CHENG JinPing, YUAN XingXing. Whole Genome Association Study of Protein and Threonine Content in Mung Bean Seeds and Development of KASP Molecular Marker[J]. Scientia Agricultura Sinica, 2026, 59(13): 2789-2801.

图/表 9

表1

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表3

显著QTL关联候选基因的分析"

染色体 Chromosome	位置 Position (bp)	基因 Gene	拟南芥同源基因 Homologous gene of Arabidopsis	功能注释 Functional annotation	参考文献 References
LG03	880773-881672	EVM0013103	AT3G09110	非特征蛋白LOC106770734 Uncharacterized protein LOC106770734
LG03	899614-909356	EVM0002414	AT3G55320	ABC转运蛋白B家庭成员20 ABC transporter B family member 20
LG03	910014-911736	EVM0023300	AT2G39470	叶绿体腔定位光合NDH亚基1 Photosynthetic NDH subunit of lumenal location 1, chloroplastic
LG03	914126-915697	EVM0026632	AT2G39490	F-box蛋白At2g39490 F-box protein At2g39490
LG03	916012-918173	EVM0019126	AT3G55340	核仁素2 Nucleolin 2
LG03	921375-922998	EVM0031542	AT5G59790	类FLC上游蛋白 Protein upstream of FLC-like
LG03	925340-928379	EVM0016950	AT3G55350	类ALP蛋白 Protein ALP1-like
LG03	929960-930569	EVM0003649	AT2G39500	非特征蛋白LOC108325383，部分 Uncharacterized protein LOC108325383, partial
LG03	937645-938435	EVM0002963	AT2G37430	类锌指蛋白ZAT11 Zinc finger protein ZAT11-like
LG03	943232-949840	EVM0000757	AT5G65750	2-酮戊二酸脱氢酶E1组分 2-Oxoglutarate dehydrogenase E1 component	[27]
LG03	950896-951478	EVM0004529	-	非特征蛋白类At4g04775 Uncharacterized protein At4g04775-like
LG03	952343-952875	EVM0028246	AT3G05850	非特征蛋白LOC106770420 Uncharacterized protein LOC106770420
LG03	959895-962941	EVM0006824	AT5G59840	ras相关蛋白RABE1c ras-related protein RABE1c
LG03	964662-966659	EVM0003873	AT4G35550	WUSCHEL相关同源盒13 WUSCHEL-related homeobox 13
LG03	970064-975466	EVM0008260	AT5G13680	延伸复合蛋白1亚型X1 Elongator complex protein 1 isoform X1
LG03	977401-984719	EVM0013687	AT3G55400	甲硫氨酸-tRNA连接酶，叶绿体/线粒体 Methionine--tRNA ligase, chloroplastic/mitochondrial
LG03	985197-986871	EVM0005089	-	假设蛋白VIGAN_08013300，部分 Hypothetical protein VIGAN_08013300, partial
LG03	988811-990132	EVM0031142	AT3G55390	类CASP蛋白4C2 CASP-like protein 4C2

表3

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性状 Trait	最大值 Maximum	最小值 Minimum	平均数±标准差 Mean±SD	变异系数 CV (%)	偏度系数 Skewness	峰度系数 Kurtosis	遗传力 H² (%)
蛋白质Protein (%)	24.38	18.45	21.27±1.01	4.75	0.06	0.09	76.38
苏氨酸Threonine (PA)	3601595.23	834158.43	1509441.75±410080.96	27.17	1.14	2.60	-
PA：峰面积 PA: Peak area

序号Number	位点SNP	染色体Chromosome	位置Position (bp)	-log₁₀_(P)	表型贡献率R² (%)
1	SLG03_939296	3	939296	5.64	6.16
2	SLG03_942729	3	942729	6.29	6.79
3	SLG03_942730	3	942730	6.29	6.79
4	SLG03_1038622	3	1038622	5.16	6.06
5	SLG03_966208	3	966208	7.01	11.21
6	SLG03_1039749	3	1039749	7.70	12.20

绿豆籽粒蛋白质与苏氨酸含量全基因组关联分析及KASP分子标记开发

Whole Genome Association Study of Protein and Threonine Content in Mung Bean Seeds and Development of KASP Molecular Marker

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