Mining SSR Loci and Analysis the Genetic Diversity of Tartary Buckwheat Based on the Whole Genome Sequence

doi:10.13304/j.nykjdb.2021.0521

Abstract

Abstract:

Tartary buckwheat （Fagopyrum tataricum） is an important minor crop. In this study， molecular markers of the simple sequence repeat （SSR） were mined from 8 chromosomes sequence of tartary buckwheat in GenBank database. SSR primers were designed by primer 3.0 software and those with high polymorphism were employed to evaluate the genetic diversity. A total of 51 485 SSR loci were screened with average frequency of 114.07 Mb^-1. Among these SSR motifs， dinucleotide repeat was the main type accounting for 78.20%， followed by trinucleotide repeat （17.76%）. There were 361 types of repeat motifs， while AT/AT （69.60 %）， AAT/TTA （2.49%） and AGA/TCT （2.10%） were the main repeat types. The SSRs length ranged from 12 to 228 bp， and the ratio of SSRs with length ranging from 12 to 20 bp was 57.93%， the ratio with length above 20 bp was 42.07%. It was found that the SSR types detected were specific to the chromosomes. Based on the type of SSR loci， 156 pairs of primers were designed and 17 pairs of primers were screened with high polymorphism to analyze the genetic diversity of the 42 local varieties of tartary buckwheat. The diversity of black tartary buckwheat samples in Meigu County was the highest. The diversity of samples in Butuo County was significantly lower than that in the other two counties. It was of great significance to identify and analyze tartary buckwheat germplasm resources and molecular marker-assisted breeding through a large number of SSR mining， especially the chromosome specific SSR loci mining. In this study， a large number of SSRs were mined， especially the chromosomal specific SSR sites were mined， which was of great significance for the identification and analysis of tartary buckwheat germplasm resources and the molecular marker-assisted breeding of tartary buckwheat. Genetic diversity analysis also revealed the availability of SSR markers and the level of genetic diversity of tartary buckwheat， which provided a new approach for the study of the conservation of genetic diversity of tartary buckwheat.

Key words: Fagopyrum tataricum, chromosome, simple repeat sequence （SSR）, genetic diversity

摘要：

苦荞是一种重要的杂粮作物。利用GenBank数据库中苦荞的8条染色体基因组序列进行SSR标记挖掘，并基于Primer 3.0设计SSR引物，筛选多态性高的引物进行遗传多样性评价。共检测到51 485个SSR 位点，平均发生频率为114.07 Mb^-1，二核苷酸重复型最多（78.20%），其次为三核苷酸重复型（17.76%）。苦荞SSR中包含361种类型重复基元，具有碱基偏好性，优势基元为AT/TA （69.60%）、AAT/TTA （2.49 %）和AGA/TCT（2.10 %）。序列长度变化范围为12~228 bp，长度12~20 bp的占比57.93 %，长度大于20 bp 的占比42.07 %。SSR位点分布在每条染色体上的数量和种类特征较为一致，不同染色体能检出的SSR种类具有特异性。根据不同类型SSR位点设计并合成156对引物，筛选出17对多态性较高的引物用于遗传多样性分析，发现42份苦荞地方品种资源具有较高的多样性，美姑县的黑苦荞遗传多样性最高，布拖县的样品多数聚为同一分支，多样性显著低于美姑县和昭觉县的品种，西南种质库保存的野生苦荞与其他栽培品种明显分离。通过大量SSR的挖掘特别是染色体特异性SSR位点的挖掘，对苦荞种质资源的鉴定分析以及苦荞分子标记辅助育种有重要意义；遗传多样性检测也显示了SSR分子标记的可用性，以及黑苦荞的遗传多样性水平，对苦荞遗传多样性保护研究提供了新的途径。

关键词: 苦荞, 染色体, 简单重复序列（SSR）, 遗传多样性

CLC Number:

S517

Xixi ZUO, Yingjie SONG, Xinyan MA, Yunhui YANG, Yifei WANG, Zeguang GUO, Xiongzhi ZHU, Yue LIU. Mining SSR Loci and Analysis the Genetic Diversity of Tartary Buckwheat Based on the Whole Genome Sequence[J]. Journal of Agricultural Science and Technology, 2022, 24(4): 38-51.

左茜茜, 宋英杰, 马心妍, 杨云卉, 王轶菲, 郭泽光, 朱雄智, 刘越. 苦荞全基因组SSR位点挖掘及遗传多样性分析[J]. 中国农业科技导报, 2022, 24(4): 38-51.

Figures/Tables 12

References 43

1	林汝法.中国荞麦[M].北京:中国农业出版社, 1994:1-251.
2	CHEN Q F. A study of resource of Fagopyrum (Polygonaceae) native to China [J]. Bot. J. Linn. Soc., 1999, 130(1):53-64.
3	赵钢,唐宇,王安虎.发展中国的苦荞生产[J].作物杂志, 2002, 18(4):11-12.
4	屈洋,周瑜,王钊,等.苦荞产区种质资源遗传多样性和遗传结构分析[J].中国农业科学, 2016, 49(11):2049-2062.
	QU Y, ZHOU Y, WANG Z, et al... Analysis of genetic diversity and structure of tartary buckwheat resources from production regions [J]. Sci. Agric. Sin., 2016, 49(11):2049-2062.
5	杨明君,郭忠贤,杨媛,等.我国荞麦种植简史[J].内蒙古农业科技, 2007, (5):85-86.
6	范昱.中国苦荞种质资源性状评价和荞麦属植物亲缘关系分析[D].成都:成都大学, 2019.
	FAN Y. Evaluation of tartary buckwheat germplasm resources in China and genetic relationship analysis of Fagopyrum Mill genus plants [D]. Chengdu: Chengdu University, 2019.
7	吴韬,肖丽,李伟丽.苦荞的营养与功能成分研究进展[J].西华大学学报(自然科学版), 2021, 40(2):91-109.
	WU T, XIAO L, LI W L. Research progress of chemicals in tartary buckwheat [J]. J. Xihua Univ.(Nat. Sci.), 2021, 40(2):91-109.
8	王亦君,冯舒涵,程锦堂,等.大黄蒽醌类化学成分和药理作用研究进展[J].中国实验方剂学杂志, 2018, 24(13):227-234.
	WANG Y J, FENG S H, CHENG J T, et al.. Research progresson chemical constituents and pharmacological action of anthraquinone in Rhei Radix et Rhizoma [J]. Chin. J. Exp. Traditional Medical Formulae, 2018, 24(13):227-234.
9	WU X Q, GE X S, LIANG S X, et al.. A novel selective accelerated solvent extraction for effective separation and rapid simultaneous determination of six anthraquinones in tartary buckwheat and its products by UPLC-DAD [J]. Food Anal. Methods, 2015, 8(5):1124-1132.
10	祝寅淏,王帅,李瑶,等.黄酮类化合物药理作用的研究进展[J].吉林医药学院学报, 2018, 39(3):219-223.
11	ZHANG L, LI X, MA B, et al.. The tartary buckwheat genome provides insights into rutin biosynthesis and abiotic stress tolerance [J]. Mol. Plant, 2017, 10(9):1224-1237.
12	DING S, WANG S, HE K, et al.. Large-scale analysis reveals that the genome features of simple sequence repeats are generally conserved at the family level in insects [J]. BMC Genomics, 2017, 18(1):848.
13	马名川,刘龙龙,刘璋,等.苦荞全基因组 SSR 位点特征分析与分子标记开发[J].作物杂志, 2021(1):38-46.
	MA M C, LIU L L, LIU Z, et al... Analysis of SSR lci in whole genome and development of molecular markers in tartary buckwheat [J]. Crops, 2021(1):38-46.
14	杜伟,王东航,侯思宇,等.基于苦荞全长转录组测序开发SSR标记及遗传多样性分析[J]. 植物生理学报, 2020, 56 (7):1432-1444.
	DU W, WANG D H, HOU S Y, et al... Development of SSR markers based on full-length transcriptome sequencing and its application for genetic diversity analysis in Fagopyrum tataricum [J]. Plant Physiol. J., 2020, 56(7):1432-1444.
15	贺润丽,尹桂芳,李春花,等.苦荞种皮转录组SSR位点信息分析及其分子标记的开发[J]. 分子植物育种, 2020, 18(18):213-220.
	HE R L, YIN G F, LI C H, et al... Development of molecular markers and SSR loci information analysis of transcriptome in tartary buckwheat seed coat [J]. Mol. Plant Breeding, 2020, 18(18):213-220.
16	李春花,陈蕤坤,王艳青,等.利用SSR标记构建云南苦荞种质资源分子身份证[J].分子植物育种, 2019, 17(5):1575-1582.
	LI C H, CHEN R K, WANG Y Q, et al.. Establishment of the molecular ID for Yunnan tartary buckwheat germplasm resources based on SSR marker [J]. Mol. Plant Breeding,2019, 17(5):1575-1582.
17	黎瑞源,潘凡,陈庆富,等.苦荞转录组EST-SSR发掘及多态性分析[J].中国农业科技导报, 2015, 17(4):42-52.
	LI R Y, PAN F, CHEN Q F, et al... Excavation and polymorphism analysis of EST-SSR from transcriptome of tartary buckwheat [J]. J. Agric. Sci. Technol., 2015, 17(4):42-52.
18	韩瑞霞,张宗文,吴斌,等.苦荞SSR引物开发及其在遗传多样性分析中的应用[J].植物遗传资源学报, 2012, 13(5):759-764.
	HAN R X, ZHANG Z W, WU B, et al.. Development of SSR markers and application in analysis of genetic diversity in tartary buckwheat(Fagopyrum tataricum)[J]. J. Plant Genetic Resour., 2012, 13(5):759-764.
19	刘越,范增华,孙洪波,等.裂叶牵牛获得表达序列标签资源的简单序列重复信息分析[J]. 中国药学杂志, 2011, 46(23):1790-1794.
	LIU Y, FAN Z H, SUN H B, et al.. Analysis of SSR information in EST resource of Pharbitis nil [J]. Chin. Pharm. J., 2011, 46(23):1790-1794.
20	TEMNYKH S, PARK W D, AYRES N, et al... Mapping and genome organization of microsatellite sequences in rice (Oryza sativa L.) [J]. Theor. Appl. Genet., 2000, 100(5):697-712.
21	ZHANG P, DREISIGACKER S, MELCHINGER A E, et al.. Quantifying novel sequence variation and selective advantage in synthetic hexaploid wheats and their backcross-derived lines using SSR markers [J]. Mol. Breeding, 2005, 15(1):1-10.
22	HOU S Y, SUN Z X, HU B L, et al.. Genetic diversity of buckwheat cultivars (Fagopyrum tartaricum Gaertn.) assessed with SSR markers developed from genome survey sequences [J]. Plant Mol. Biol. Rep., 2016, 34:233-241.
23	FANG X M, HUANG K H, NIE J, et al.. Genome-wide mining, characterization, and development of microsatellite markers in Tartary buckwheat (Fagopyrum tataricum Garetn.) [J]. Euphytica, 2019, 215:183-193.
24	VARSHNEY R K, GRANER A, SORRELLS M E. Genic microsatellite markers in plants:features and applications [J]. Trends Biotechnol., 2005, 23(1):48-55.
25	ZHAO H S, YANG L, PENG Z H, et al.. Developing genome-wide microsatellite markers of bamboo and their applications on molecular marker assisted taxonomy for accessions in the genus Phyllostachys [J/OL]. Sci. Rep., 2015, 5:8018 [2022-01-09]..
26	李水根,关媛,李秀芬,等.观赏樱属植物基因组SSR分子标记开发及鉴定[J/OL].分子植物育种,2021 [2022-01-09]..
	LI S G, GUAN Y, LI X F, et al.. Development and characterization of genome-SSR markers in flowering cherries [J/OL]. Mol. Plant Breeding, 2021 [2022-01-09]. .
27	张曼,田娟,孙墨可,等.基于向日葵全基因组序列的SSR标记开发及鉴定[J/OL].分子植物育种, 2021 [2022-01-09]. .
	ZHANG M, TIAN J, SUN M K, et al.. Development of SSR molecular markers based on whole genome of Sunflower [J/OL]. Mol. Plant Breeding, 2021 [2022-01-09]. .
28	何志凯,耿平美惠,张勇洪,等.药用植物益智的基因组调研及SSR分子标记开发[J/OL].分子植物育种, 2021 [2022-01-09]. .
	HE Z K, GENG P M H, ZHANG Y H, et al.. Genome survey and SSR molecular marker development of medicinal plant Alpinia oxyphylla Miq [J/OL]. Mol. Plant Breeding, 2021 [2022-01-09]. .
29	周文才,唐山,何小三,等.光皮树基因组SSR特征分析[J/OL].分子植物育种,2021 [2022-01-09]..
	ZHOU W C, TANG S, HE X S, et al.. Analysis of SSR Characteristics in Genome of Swida wilsoniana [J/OL]. Mol. Plant Breeding,2021 [2022-01-09]. .
30	张淑文,梁森苗,郑锡良,等. 杨梅基因组SSR引物的开发与应用[J].园艺学报, 2019, 46(1):149-156.
	ZHANG S W, LIANG S M, ZHENG X L, et al.. Development of genomic SSR and application in Chinese bayberry [J]. Acta Hortic. Sin., 2019, 46(1):149-156.
31	原志敏.玉米全基因组SSRs分子标记开发与特征分析[D].四川雅安:四川农业大学, 2013.
	YUAN Z M. Development and characterization of SSR markers providing genome-wide coverage and High Resolution in Maize [D]. Sichuan Yaan: Sichuan Agricultural University, 2013.
32	石桃雄,黎瑞源,郭菊卉,等.基于普通荞麦种子表达序列标签微卫星标记的开发[J].贵州农业科学, 2014, 42(3):1-5.
	SHI T X, LI R Y, GUO J H, et al.. Development of SSR molecular markers based on expressed sequence tags from seeds of Fagopyrum esculentum [J]. Guizhou Agric. Sci., 2014, 42(3):1-5.
33	蒋爽,张学英,安海山,等.枇杷全基因组SSR标记开发及其多态性研究[J].园艺学报,2021(5):170-179.
	JIANG S, ZHANG X Y, AN H S, et al.. A study on the development and analysis of polymorphism of SSR markers in the whole genome of Loquat [J]. Acta Hortic, Sinica, 2021(5):170-179.
34	乔舒婷,董文其,胡齐赞,等.基于丝瓜全基因组序列SSR分子标记开发[J/OL].分子植物育种,2021 [2022-01-09]. .
	QIAO S T, DONG W Q, HU Q Z, et al.. Development of SSR molecular markers based on whole genome sequences of sponge gourd [J/OL]. Mol. Plant Breeding, 2021 [2022-01-09]. .
35	詹海仙,王颖莉,杜晨晖,等.基于甘草全基因组序列的SSR分子标记开发[J].分子植物育种, 2020, 18(18):221-228.
	ZHAN H X, WANG Y L, DU C H, et al.. SSR molecular markers development based on whole genome sequences in Glycyrrhiza uralensis Fisch [J]. Mol. Plant Breeding, 2020, 18(18):221-228.
36	刘松卫,卢迎春,宋婉玲,等.基于灯盏花全基因组SSR位点分析及多态性引物开发[J].分子植物育种, 2018, 16(12):4003-4009.
	LIU S W, LU Y C, SONG W L, et al.. SSR loci analysis based on Erigeron breviscapus genome and polymorphism primers development [J]. Mol. Plant Breeding, 2018, 16(12):4003-4009.
37	洪文娟,郝兆祥,刘康佳,等.基于石榴全基因组序列的SSR标记开发及鉴定[J].北京林业大学学报, 2019, 41(8):38-47.
	HONG W J, HAO Z X, LIU K J, et al.. Development and identification of SSR molecular markers based on whole genomic sequences of Punica granatum [J]. J. Beijing Forestry Univ.,2019, 41(8):38-47.
38	SENTHILKUMARAN R, BISHT I S, BHAT K V, et al.. Diversity in buckwheat (Fagopyrum spp.) landrace populations from north-western Indian Himalayas [J]. Genetic Resour. Crop Evol., 2008, 55(2):287-302.
39	BOTSTEIN D, WHITE R L, SKOLNICK M, et al.. Construction of a genetic linkage map in man using restriction fragment length polymorphisms [J]. Am. J. Human Genetics, 1980, 32:314-331.
40	高帆,张宗文,吴斌.中国苦荞SSR分子标记体系构建及其在遗传多样性分析中的应用[J]. 中国农业科学, 2012, 45(6):1042-1053.
	GAO F, ZHANG Z W, WU B. Construction and application of SSR molecular markers system for genetic diversity analysis of Chinese tartary buckwheat germplasm resources [J]. Sci. Agric. Sin., 2012, 45(6):1042-1053.
41	杨学文,丁素荣,胡陶,等.104份苦荞种质的遗传多样性分析[J].作物杂志, 2013 (6):13-18.
	YANG X W, DING S R, HU T, et al.. Genetic diversity of 104 tartary buckwheat accessions [J]. Crops, 2013 (6):13-18.
42	张久盘,常克勤,杨崇庆,等.基于ITS和RLKs序列的苦荞种质资源遗传多样性分析[J].南方农业, 2020, 14(3):157-160.
43	SONG Y J, FANG Q, JARVIS D, et al.. Network analysis of seed flow, a traditional method for conserving tartary buckwheat (Fagopyrum tataricum) landraces in Liangshan, Southwest China [J/OL]. Sustainability, 2019, 11(16):4263 [2021-01-09]. .

采集地点 Sampling site	样品编号 Sample ID	来源 Source	数量 Amount
四川攀枝花 Panzhihua， Sichuan	XNZY-1	中国西南野生生物种质资源库 Germplasm Bank of Wild Species in Southwest China	3
西藏 Tibet	XNZY-2
河北 Hebei	XNZY-5
贵州 Guizhou	GZSF-1、GZSF-3、GZSF-4	贵州省荞麦工程技术研究中心 Guizhou Province Buckwheat Engineering Technology Research Center	4
湖南 Hunan	GZSF-2		4
山西 Shanxi	DTZX-1	云南省滇台特色农业产业化工程研究中心 Yunnan-Taiwan Engineering Research Center for Characteristic Agriculture Industrialization of Yunnan Province	3
云南 Yunnan	DTZX-2
重庆 Chongqing	DTZX-5
四川省昭觉县 Zhaojue， Sichuan	ZJJZ-1、ZJJZ-2、ZJLG、ZJSJ、ZJSY-1、ZJSY-2、ZJWQ-1、ZJWQ-2、ZJWQ-3、ZJWZ、ZJXY、EN-G-TU、CHU-G	本课题组采集 Sampled by this lab	13
四川省美姑县 Meigu， Sichuan	MG-48、MGLL、MGNY-1、MGNY-2、MGTZ、MGYD-1、MGYD-2、MGYS、EN-G-JIE、EN-G-WAZI	本课题组采集 Sampled by this lab	10
四川省布拖县 Butuo， Sichuan	BT-52、BTAE、BTDM、BTLC、BTLG、BTLJ、BTLK-1、BTLK-2、BTZE	本课题组采集 Sampled by this lab	9
总计Total			42

采集地点 Sampling site	样品编号 Sample ID	来源 Source	数量 Amount
四川攀枝花 Panzhihua， Sichuan	XNZY-1	中国西南野生生物种质资源库 Germplasm Bank of Wild Species in Southwest China	3
西藏 Tibet	XNZY-2
河北 Hebei	XNZY-5
贵州 Guizhou	GZSF-1、GZSF-3、GZSF-4	贵州省荞麦工程技术研究中心 Guizhou Province Buckwheat Engineering Technology Research Center	4
湖南 Hunan	GZSF-2		4
山西 Shanxi	DTZX-1	云南省滇台特色农业产业化工程研究中心 Yunnan-Taiwan Engineering Research Center for Characteristic Agriculture Industrialization of Yunnan Province	3
云南 Yunnan	DTZX-2
重庆 Chongqing	DTZX-5
四川省昭觉县 Zhaojue， Sichuan	ZJJZ-1、ZJJZ-2、ZJLG、ZJSJ、ZJSY-1、ZJSY-2、ZJWQ-1、ZJWQ-2、ZJWQ-3、ZJWZ、ZJXY、EN-G-TU、CHU-G	本课题组采集 Sampled by this lab	13
四川省美姑县 Meigu， Sichuan	MG-48、MGLL、MGNY-1、MGNY-2、MGTZ、MGYD-1、MGYD-2、MGYS、EN-G-JIE、EN-G-WAZI	本课题组采集 Sampled by this lab	10
四川省布拖县 Butuo， Sichuan	BT-52、BTAE、BTDM、BTLC、BTLG、BTLJ、BTLK-1、BTLK-2、BTZE	本课题组采集 Sampled by this lab	9
总计Total			42

重复基元 Repeat motif	Chr.1	Chr.2	Chr.3	Chr.4	Chr.5	Chr.6	Chr.7	Chr.8	总计 Total	比例 Percentage/%
AT/TA	5 194	4 853	4 566	4 432	4 260	4 606	4 019	3 901	35 831	69.6
CT/GA	280	310	254	347	299	278	225	216	2 209	4.29
AG/TC	184	152	158	178	141	133	122	147	1 215	2.36
AC/TG	137	124	99	119	83	111	92	105	870	1.69
CA/GT	95	94	78	114	83	66	61	81	672	1.31
AAT/TTA	180	171	183	144	160	144	155	144	1 281	2.49
AGA/TCT	169	183	104	146	112	133	118	116	1 081	2.10
ATA/TAT	174	129	144	104	140	122	104	135	1 052	2.04
AAG/TTC	139	120	104	120	100	108	85	85	861	1.67
CTT/GAA	129	99	89	103	101	73	88	100	782	1.52
ATT/TAA	142	93	103	82	105	86	91	76	778	1.51
CAT/GTA	55	48	24	55	41	38	28	41	330	0.64
CAA/GTT	54	33	36	34	36	31	27	30	281	0.55
ACT/TGA	35	39	31	25	34	25	30	36	255	0.50
AAAT/TTTA	60	45	0	35	42	39	48	40	309	0.60
AATA/TTAT	27	23	35	33	20	19	26	20	203	0.39
AAGA/TTCT	26	17	17	23	20	30	17	13	163	0.32
ATAA/TATT	29	19	24	16	19	0	17	25	149	0.29
ATTT/TAAA	18	9	22	12	16	7	21	20	125	0.24
AAAAT/TTTTA	3	9	2	0	3	5	5	3	30	0.06
AGAAG/TCTTC	0	7	3	3	0	10	6	4	27	0.05
AAATA/TTTAT	7	0	0	0	0	0	3	3	13	0.03
AAAATA/TTTTAT	0	0	0	0	0	2	0	0	2	≈0.00
TCTACC	1	0	1	0	1	0	1	0	4	≈0.00
GTTCAG	1	1	0	1	1	0	0	0	4	≈0.00
总计Total	7 139	6 578	6 077	6 126	5 817	6 066	5 389	5 341	48 527	—

重复基元 Repeat motif	Chr.1	Chr.2	Chr.3	Chr.4	Chr.5	Chr.6	Chr.7	Chr.8	总计 Total	比例 Percentage/%
AT/TA	5 194	4 853	4 566	4 432	4 260	4 606	4 019	3 901	35 831	69.6
CT/GA	280	310	254	347	299	278	225	216	2 209	4.29
AG/TC	184	152	158	178	141	133	122	147	1 215	2.36
AC/TG	137	124	99	119	83	111	92	105	870	1.69
CA/GT	95	94	78	114	83	66	61	81	672	1.31
AAT/TTA	180	171	183	144	160	144	155	144	1 281	2.49
AGA/TCT	169	183	104	146	112	133	118	116	1 081	2.10
ATA/TAT	174	129	144	104	140	122	104	135	1 052	2.04
AAG/TTC	139	120	104	120	100	108	85	85	861	1.67
CTT/GAA	129	99	89	103	101	73	88	100	782	1.52
ATT/TAA	142	93	103	82	105	86	91	76	778	1.51
CAT/GTA	55	48	24	55	41	38	28	41	330	0.64
CAA/GTT	54	33	36	34	36	31	27	30	281	0.55
ACT/TGA	35	39	31	25	34	25	30	36	255	0.50
AAAT/TTTA	60	45	0	35	42	39	48	40	309	0.60
AATA/TTAT	27	23	35	33	20	19	26	20	203	0.39
AAGA/TTCT	26	17	17	23	20	30	17	13	163	0.32
ATAA/TATT	29	19	24	16	19	0	17	25	149	0.29
ATTT/TAAA	18	9	22	12	16	7	21	20	125	0.24
AAAAT/TTTTA	3	9	2	0	3	5	5	3	30	0.06
AGAAG/TCTTC	0	7	3	3	0	10	6	4	27	0.05
AAATA/TTTAT	7	0	0	0	0	0	3	3	13	0.03
AAAATA/TTTTAT	0	0	0	0	0	2	0	0	2	≈0.00
TCTACC	1	0	1	0	1	0	1	0	4	≈0.00
GTTCAG	1	1	0	1	1	0	0	0	4	≈0.00
总计Total	7 139	6 578	6 077	6 126	5 817	6 066	5 389	5 341	48 527	—

SSR引物类型 SSR primer type	总引物数量 Total primers number	有效引物数量 Effective primers number	多态性引物数量 Polymorphic primers number	基序长度 Sequence length /bp
复合型 Compound	3	3	2	55、66
二核苷酸 Dinucleotide	119	86	11	14~66
三核苷酸 Trinucleotide	26	23	4	15、27、48、63
四核苷酸Tetranucleotide	7	7	—	—
五核苷酸Pentanucleotide	1	1	—	—
总计Total	156	120	—	—