Genetic Diversity and Association Analysis between Medicinal Components and ISSR Markers in Astragali Radix
MENG Xiang-shan1,2, ZHOU Yu-mei1,2, DAI Xiao-hua2, LIU Ping1,2*
1. Key Laboratory of Modern Molecular Breeding for Dominant and Special Crops in Ningxia,Yinchuan 750021, China; 2. College of Agronomy, Ningxia University,Yinchuan 750021, China
Abstract:OBJECTIVE To study and exploit Chinese medicine Astragali Radix, the molecular markers that relates to the phenotypic traits on medicinal components of Astragali Radix and would be detected. METHODS The genetic diversity of 43 Astragali Radix samples was analyzed with ISSR molecular marker technique and then the population genetic structure was studied through 13 selected markers. The association analysis between ISSR markers and 4 phenotypic traits of medicinal components were performed with GLM (general linear model) programs in Tassel 2.1. Certain genetic diversity was discovered among the 43 Astragali Radix samples. RESULTS The genetic distance varied between 0.050 6 and 0.743 8, with an average of 0.274 1. Moreover, the cultivated Astragali Radix from Ningxia and Gansu province closely related to the wild Astragali Radix collected from Liupanshan town in Ningxia. On the other hand, No. 340 had the farthest relationship with other Astragali Radix. The content of polysaccharide, total saponins, total flavonoids, and Astragaloside IV ranged between 7.693-27.840 mg·g-1, 7.167-17.579 mg·g-1, 2.212-6.164 mg·g-1 and 6.070-107.920 μg·g-1, respectively. Meanwhile, linear regression analysis indicated that there was a significant positive correlation between the content of the total saponins and that of flavonoids (r=0.650 5,P=2.3×10-6<0.01), while the content of astragaloside Ⅳ had no significant correlation with that of polysaccharide, total saponins and total flavonoids. The population genetic structural analysis showed that the 43 samples were divided into 4 subgroups. There were total of 34 locus in 13 ISSR markers significantly associated (P<0.01) with the content of polysaccharide,total saponins, flavonoids and astragaloside Ⅳ, and the rate of explanation on the phenotype of related marker ranged from 8.14% to 51.39%. Among the locus, 15 were related with astragaloside Ⅳ content at interpretation rates above 30%, 1 with polysaccharide content an interpretation rate reached as high as 51.39% with high threshold (P<1×10-5). CONCLUSION These results would provide supporting evidence for identification and protection of germplasm resources as well as molecular marker-assisted breeding.
孟祥善, 周玉梅, 代晓华, 刘萍. 黄芪ISSR标记遗传多样性及其与主要药用成分的关联分析[J]. 中国药学杂志, 2019, 54(24): 2060-2070.
MENG Xiang-shan, ZHOU Yu-mei, DAI Xiao-hua, LIU Ping. Genetic Diversity and Association Analysis between Medicinal Components and ISSR Markers in Astragali Radix. Chinese Pharmaceutical Journal, 2019, 54(24): 2060-2070.
Editorial Committee of Flora of China. Flora Reipublicae Popularis Sinicae(40 volumes)(中国植物志.第四十卷)[M]. Beijing: Science Press, 1998:80-81.
[2]
Ch.P(2010). Vol Ⅰ(中国药典2010年版.一部)[S]. 2010:283.
[3]
ZHANG Y D, WANG Y L, SHENG J P, et al. Effects on blood pressure and inflammation of astragalus saponin 1, a principle isolated from Astragalus membranaceus Bge[J]. Acta Pharm Sin(药学学报),1984,19(5):333-337.
[4]
YAN Y D,HUANG X J,YANG Y. Effect of astragaloside Ⅳ on changes of adiponectin and its receptor adipo R1 in injured myocardium in rats[J]. Chin Pharm J(中国药学杂志), 2014, 49(11):978-981.
[5]
LU Z Q. Protective efect of hydroxysafflor yellow a combined with astragaloside Ⅳ on chronic kidney disease[J]. J Nanjing Univ Tradit Chin Med(南京中医药大学学报), 2018,34(6):589-592.
[6]
XIA G Q, HAN X J.Effect of astragalus polysaccharide on cell proliferation and the relevant gene expression during senescence of zebrafish[J]. Chin Pharm J(中国药学杂志), 2012, 47(13):1039-1041.
[7]
WANG T, WEI X J, WENG X G, et al. Effects of astragalus polysaccharides on theinsulin sensitivity in the fat rats[J]. Chin Pharm J(中国药学杂志), 2011, 46(3): 185-188.
[8]
XUAN L Y, TAO X X, ZHAO Y J, et al. Effect of total flavonoids of astragalus on endoplasmic reticulum chaperone, calumenin and connecxin 43 in suckling mouse myocardium with myocarditis caused by coxsackievirus B3[J]. Chin J Appl Physiol(中国应用生理学杂志), 2016, 32(1):51-54.
[9]
YUAN H, ZHANG S F, CHEN N, et al. Progress in application and biological activity of Radix Astragali in health foods[J]. Food Sci(食品科学), 2014, 35(15):330-334.
[10]
NA H J,UM J Y,KIM S C, et al. Molecular discrimination of medicinal Astragali Radix by RAPD analysis[J].Immunopharmacol Immunotoxicol , 2004, 26(2): 265-272.
[11]
YIP P Y,KWAN H S.Molecular identification of Astragalus membranaceus at the species and locality levels[J].J Ethnopharmacol, 2006, 106(2): 222-229.
[12]
JIAO M L, LI Z Y, QIN X M, et al. Comparison between Astragalus membranaceus var. mongholicus and Hedysarum polybotrys based on ITS sequences and metabolomics[J]. Acta Pharm Sin(药学学报), 2015, 50 (12): 1625-1631.
[13]
WANG X F, LIANG Y, HU Z L, et al. Study on genetic diversity of Astragalus membranaceus var. mongholicus populations in innormongolia[J]. Grassl China(中国草地学报), 2018,40(1): 42-48.
[14]
GUO H Y, WANG W W, YU J, et al. DNA Barcoding provides distinction between Radix Astragali and its adulterants [J]. Sci China Life Sci(中国科学·生命科学),2010, 53(8): 992-999.
[15]
NORDBORG M. Linkage disequilibrium, gene trees and selfing: An ancestral recombination graph with partial self-fertilization[J]. Genets Soc Am, 2000, 154: 923-929.
[16]
FLINT-GARCIA S A, THORNSBERRY J M, BUCKLER Ⅳ E S. Structure of linkage disequilibrium in plants[J]. Annu Rev Plant Biol, 2003, 54: 357-374.
[17]
GUPTA P K, RUSTGI S, KULWAL P L. Linkage disequilibrium and association studies in higher plants: present status and future prospects[J]. Plant Mol Biol, 2005, 57: 461-485.
[18]
HANSEN M, KRAFT T, GANESTAM S, et al. Linkage disequilibrium mapping of the bolting gene in sea beet using AFLP markers[J]. Genet Res, 2001, 77(1): 61-66.
[19]
JUN T H, VAN K, KIM M Y, et al. Association analysis using SSR markers to find QTL for seed protein content in soybean[J]. Euphytica, 2008, 162: 179-191.
[20]
LAI Y, WANG P X, WANG H J, et al. Genetic diversity and association analysis using SSR markers in barley[J]. Sci Agric Sin(中国农业科学),2013, 46(2): 233-242.
[21]
LIU J R. Genetic diversity analysis of Sophora alopecuroides and the correlation analysis with matrine[D].Yinchuan: Ningxia University, 2013.
[22]
MENG X S, DAI X H, LIU P, et al. Study on genetic diversity of ITS sequence in Stellaria dichotoma var. lacceolata[J]. J Chin Med Mater(中药材),2018, 41(1):55-59.
[23]
ZHANG R. Analysis on genetic diversity of Radix Astragali by ISSR marker[D]. Jinzhong: Shanxi Agricultural University, 2014.
[24]
ZOU Q. Guidance of Plant Physiology Experiments(植物生理学实验指导)[M]. Beijing: China Agriculture Press, 2000: 110-111.
[25]
EVANNO G,REGNAUT S,GOUDET J. Detecting the number of clusters of individuals using the software structure: a simulation study[J]. Mol Ecol, 2005, 14(8):2611-2620.
[26]
YAN S, REN W C, LIU Z D, et al. Correlation analysis of levels of astraglus polysaccharides, saponin and flavone[J]. West J Tradit Chin Med(西部中医药), 2016,29(12):29-31.
[27]
ZHANG Y H, ZHANG L M, MA W, et al. Content determination of astragalus polysaccharide from astragalus in different regions [J]. Inf Tradit Chin Med(中医药信息),2013, 30(5): 69-71.
[28]
JIANG Y, CHAO R B. Comparison of contents of astragaloside Ⅳ and total saponins in Astragalus membranaceus[J]. West China J Pharm Sci(华西药学杂志), 2007,22(3):322-324.
[29]
GAO S Y, LI K, ZHANG X, et al. Quality comparison study between wild and cultivated Astragali Radix based on absolute growth years[J]. Chin Tradit Herb Drugs(中草药), 2018,49(10):2248-2257.
[30]
YAN J J, YAO M, LIAO G Z, et al. Determination of astragaloside Ⅳ in Astragalus membranaceus var. mongholicus with different growth years by HPLC[J]. Technol Innov Appl(科技创新与应用),2017, (28):85-87.
[31]
MACKAY I, POWELL W. Methods for linkage disequilibrium mapping in crops[J]. Trends Plant Sci, 2007, 12(2): 57-63.
[32]
SUN C W, ZHANG F Y, CHEN F, et al. Genome-wide association study for 13 agronomic traits reveals distribution of superior alleles in bread wheat from the Yellow and Huai Valley of China[J]. Plant Biotechnol J,2017, 15: 953-969.
[33]
ZHANG G H, GAO M G, LI S S, et al. Association analysis of yield traits with molecular markers in Huang-Huai river valley winter wheat region, China[J]. Acta Agron Sin(作物学报),2013,39(7): 1187-1199.