菜用大豆有机酸的全基因组关联分析

doi:10.7668/hbnxb.20194140

摘要/Abstract

摘要：

为了研究菜用大豆有机酸合成机制并为菜用大豆的品质改良提供一定的理论依据,以含有264份菜用大豆种质资源的自然群体为试验材料,分别在2020,2021年采用HPLC法测定该群体的酒石酸、苹果酸和柠檬酸含量,并结合该群体的基因型数据进行全基因组关联分析,鉴定与有机酸含量显著关联的SNP位点及候选基因。2020,2021年供试群体酒石酸平均含量分别为4.13,4.16 mg/g,苹果酸平均含量分别为7.26,8.99 mg/g,柠檬酸平均含量分别为7.12,10.88 mg/g。相关性分析发现,264份材料柠檬酸含量在2020及2021年2 a间呈显著正相关。同时,苹果酸与柠檬酸间呈显著的正相关,相关系数为0.790^*,酒石酸与苹果酸、柠檬酸均呈显著正相关,相关系数分别为0.695^*,0.739^*。结果表明,供试群体不同品种间存在较大差异,具有丰富的遗传多样性,筛选出6份具有较高有机酸含量的特异种质资源,为菜用大豆有机酸品种改良育种提供优秀的材料。基于混合线性模型的GWAS分析,共检测到54,189,43个分别与酒石酸、苹果酸及柠檬酸含量显著相关的SNP位点,同时根据基因功能注释信息,鉴定到3个及5个分别与酒石酸、苹果酸含量显著相关的候选基因。

关键词: 菜用大豆, SNP, 全基因组关联分析, 有机酸, HPLC

Abstract:

In order to study the organic acid synthesis mechanism of vegetable soybean and provide a theoretical basis for the quality improvement of vegetable soybean,a natural population containing 264 accessions of vegetable soybean was used as experimental materials.The contents of tartaric acid,malic acid and citric acid in this population were determined by high performance liquid chromatography(HPLC)in 2020 and 2021,respectively,and genome-wide association study was conducted based on the genotype data of this population,SNP loci and candidate genes significantly associated with organic acid content were identified.The results showed that the average content of tartaric acid in 2020 and 2021 populations were 4.13,4.16 mg/g,respectively;the average content of malic acid was 7.26,8.99 mg/g,respectively;the average content of citric acid was 7.12,10.88 mg/g,respectively.Correlation analysis showed that citric acid content of 264 materials showed a significant positive correlation between 2020 and 2021.Significant positive correlation coefficients were obtained for the pairs of malic/citric acids,tartaric/malic acids,and tartaric/citric acids were 0.790^*,0.695^*,0.739^*,respectively.The results showed that there were significant differences among different varieties in the test population,with rich genetic diversity.Six specific germplasm resources with high organic acid contents were selected to provide excellent materials for the improvement and breeding of organic acid varieties of vegetable soybean.Genome-wide association study based on the mixed linear model the number of SNP loci significantly correlated with the contents of tartaric acid,malic acid and citric acid were 54,189,43,respectively,and based on the gene functional annotation information,3 and 5 candidate genes were identified which were significantly correlated with the content of tartaric acid and malic acid,respectively.

Key words: Vegetable soybean, SNP, Genome-wide association study, Organic acid, HPLC

刘慧, 许文静, 杨硕, 张威, 张红梅, 刘晓庆, 朱月林, 陈华涛. 菜用大豆有机酸的全基因组关联分析[J]. 华北农学报, 2023, 38(4): 74-82. doi: 10.7668/hbnxb.20194140.

LIU Hui, XU Wenjing, YANG Shuo, ZHANG Wei, ZHANG Hongmei, LIU Xiaoqing, ZHU Yuelin, CHEN Huatao. Genome-Wide Association Study of Organic Acids in Vegetable Soybean[J]. Acta Agriculturae Boreali-Sinica, 2023, 38(4): 74-82. doi: 10.7668/hbnxb.20194140.

http://www.hbnxb.net/CN/Y2023/V38/I4/74

图/表 14

参考文献 20

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时间/min Time	甲醇/% MeOH	0.1%磷酸水溶液/% 0.1% PBS
0	2.5	97.5
10	2.5	97.5
12	100.0	0.0
17	100.0	0.0
20	2.5	97.5
25	2.5	97.5

时间/min Time	甲醇/% MeOH	0.1%磷酸水溶液/% 0.1% PBS
0	2.5	97.5
10	2.5	97.5
12	100.0	0.0
17	100.0	0.0
20	2.5	97.5
25	2.5	97.5

有机酸 Organic acid	回归方程 Linear regression equation	相关系数(R²) Coefficient of determination	定量限/(μg/mL) Quantification limit
酒石酸 Tartaric acid	y=1 453.8x-6 643.1	0.998 2	2.5
苹果酸 Malic acid	y=715.7x-4 082.6	0.997 9	3.0
柠檬酸 Citric acid	y=1 005.8x-846.9	0.999 1	10.0

有机酸 Organic acid	回归方程 Linear regression equation	相关系数(R²) Coefficient of determination	定量限/(μg/mL) Quantification limit
酒石酸 Tartaric acid	y=1 453.8x-6 643.1	0.998 2	2.5
苹果酸 Malic acid	y=715.7x-4 082.6	0.997 9	3.0
柠檬酸 Citric acid	y=1 005.8x-846.9	0.999 1	10.0

性状 Trait	环境 Environment	最小值/ (mg/g) Min	最大值/ (mg/g) Max	均值/ (mg/g) Mean	中位数 Median	极差 Mean	标准差 s	变异系数/% CV
酒石酸	2020	1.66	9.19	4.13	3.88	7.53	1.29	31.25
Tartaric acid	2021	0.48	13.52	4.16	4.03	13.04	2.17	52.07
苹果酸	2020	1.47	36.46	7.26	6.26	34.98	4.89	67.29
Malic acid	2021	1.89	25.19	8.99	8.75	23.30	4.23	47.07
柠檬酸	2020	2.56	12.05	7.12	7.09	9.49	1.37	19.22
Citric acid	2021	1.54	18.31	10.88	11.09	16.77	2.87	26.35

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