基于荧光SSR构建中国糜子核心种质DNA分子身份证

doi:10.3864/j.issn.0578-1752.2023.12.002

Abstract

Abstract:

【Objective】As an ancient minor grain crop, broomcorn millet ( Panicum miliaceum L. ) is abundant in germplasm. The construction of their DNA molecular identity based on fluorescent SSR markers would provide theoretical basis and molecular detection tool for digital management of resources. 【Method】Two hundred and thirty five broomcorn millet core accessions from China were used as experimental material, polymerase chain reaction were conducted several times using the broomcorn millet specific SSR markers which developed previously by the Broomcorn Millet Crop Molecular Breeding Research Group of the Agronomy College in Shanxi Agricultural University, core markers were obtained. With the given reference genome information of broomcorn millet, the core markers were mapped on chromosomes through BLAST sequence alignment. Fluorescence (FAM/HEX) was labeled on the 5' end of the SSR primer, the genotype of the material was given by capillary electrophoresis. Using binary coding means of expression, “0, 1” was written representing the presence or absence of amplified bands, and the discrimination of the material was detected by the software ID Analysis 4.0. Decimal (0-9) coding methods were used to calculate the size of the amplified fragments so as to obtain the character string molecular identity card of the accession. Genetic diversity, genetic clustering and principal component analysis were performed using the softwares Popgene, Powermarker, MEGA and NTSYS. The two-dimensional code DNA molecular identity card of the accession was given using the two-dimensional code online software (https://cli.im/). 【Result】PCR amplification results showed that all the 235 accessions could be separated by 7 fluorescent SSR markers (RYW3, RYW6, RYW11, RYW18, RYW37, RYW43 and RYW125) combined together. BLAST results showed that RYW18 and RYW37 were distributed on Chromosome 2, located at 0.60 cM and 0.80 cM, respectively. RYW125 is located on Chromosome 4 at 10.40 cM. RYW43 and RYW6 were distributed on Chromosome 5, located at 52.80 cM and 53.00 cM, respectively. RYW11 and RYW3 were located on Chromosome 6 at 2.10 cM and 20.70 cM, respectively. Genetic diversity analysis showed that 87 alleles were detected at 7 loci among all accessions, 3 (RYW11)-25 (RYW6) alleles were detected at each locus, with an average of 12.4286. Shannon diversity index (I) was detected and ranged from 0.2055 (RYW18) to 2.0587 (RYW6), with an average of 1.1398. The observed heterozygosity (Ho) was 0.0086 (RYW11)-0.9455 (RYW18). The expected observed heterozygosity (He) was 0.0795 (RYW18)-0.7469 (RYW11). Nei’s gene diversity index (Nei) was 0.0793 (RYW18)-0.7452 (RYW6). The polymorphism information content (PIC) was 0.0334 (RYW11)-0.8071 (RYW6), with an average of 0.5185. The results of cluster analysis and principal component analysis showed that 235 accessions were classified into 8 groups. The electrophoretic bands were number coding, and 7 marker combinations were used to construct the character string and two-dimensional code DNA molecular ID of all the accessions.【Conclusion】Two hundred and thirty five broomcorn millet core germplasms from China were used as material, polymerase chain reaction and capillary electrophoresis were conducted, 7 core SSR markers were screened. With the given reference genome information of broomcorn millet, the above markers were mapped on 4 chromosomes. Used the above SSR markers, genetic diversity analysis of all accessions was conducted and genetic diversity parameters were obtained. Based on Cluster analysis, all accessions were classified into 8 groups. Principal component analysis result resolved the deviation occured in Cluster analysis. According to the principle of most accessions were tell apart using the least markers, decimal (0-9) coding methods were used to calculate the size of the amplified fragments so as to obtain the character string molecular identity card of the accession. Combined the phenotype data with the above character string, two-dimensional code DNA molecular ID of all the accessions were developed.

Key words: broomcorn millet, capillary electrophoresis, fluorescent SSR, DNA molecular identification

XUE YaPeng, DING YiBing, WANG YuZhuo, WANG XiaoDan, CAO XiaoNing, SANTRA Dipak K, CHEN Ling, QIAO ZhiJun, WANG RuiYun. Construction of DNA Molecular Identity Card of Core Germplasm of Broomcorn Millet in China Based on Fluorescence SSR[J].Scientia Agricultura Sinica, 2023, 56(12): 2249-2261.

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Figures/Tables 10

Table 1

Table 2

Fig. 1

Fig. 2

Table 3

Amplification results and code typing of 7 pairs of core primers detected with capillary electrophoresis"

引物 Primer	荧光基团 Fluorescent group	电泳条带数目 Polymorphism bands	电泳片段长度及编码 Length and code of electrophoresis fragments (bp)
RYW3	6-羧基荧光素 FAM (blue)	19	149(01),440(02),132/149(03),132/149/156(04),138/149/178(05),143/149/156(06),143/149(07),143/149/178(08),143/149/216(09),149/156(10),149/156/216(11),149/169/198(12),149/178(13),149/211/216(14),149/216(15),156/169(16),272/344/440/474(17),440/474(18),143/149/184(19)
RYW6	6-羧基荧光素 FAM (blue)	38	242(01),255(02),262(03),300(04),308(05),323(06),333(07),387(08),444(09),452(10),174/195/242/372(11),187/195/242/372(12),195/202/242(13),195/202/242/372(14),195/242(15),195/242/267/372(16),195/242/347/372(17),195/242/372(18),195/242/387(19),195/242/401(20),202/333(21),211/242/372(22),211/333(23),242/323(24),242/333(25),242/387(26),242/425(27),252/257(28),255/267(29),296/308(30),300/308(31),323/328(32),323/333(33),323/338(34),328/333(35),333/338(36),387/425(37),444/452(38)
RYW11	6-羧基荧光素FAM (blue)	3	180(01),222(02),213/222(03)
RYW18	6-羧基荧光素FAM (blue)	5	376(01),369/381(02),376/381(03),381/386(04),381(05)
RYW37	6-羧基荧光素 FAM (blue)	6	203(01),209(02),196/203(03),196/203/209(04),196/203/209/216(05),203/209(06)
RYW125	6-羧基荧光素 FAM (blue)	24	196(01),202(02),207(03),292(04),333(05),105/202/323(06),122/211/292(07),130/151/202/333(08),130/202/219/339(09),130/202/323/333(10),130/323(11),141/259/292/372(12),141/299/323(13),196/202(14),196/219(15),202/207(16),202/211(17),202/227(18),219/223(19),259/323/333/393(20),292/333(21),299/339(22),323/339(23),323/402(24)
RYW43	六氯-6-甲基荧光素 HEX (green)	30	109(01),120(02),128(03),181(04),204(05),109/115(06),109/115/181(07),109/120/128/147(08),109/128/181/204(09),109/128(10),109/128/181(11),109/138(12),109/138/181(13),109/155/181(14),109/181(15),109/181/204(16),109/181/226/293(17),109/181/293(18),109/181/293/405 (19),109/226/293(20),109/226/293/491(21),109/226/483(22),109/226/491(23),109/293(24),128/181(25),138/181(26),170/181(27),181/204(28),103/109/181/204(29),109/204(30)

Table 3

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References 39

[1]	王星玉, 王纶. 黍稷种质资源描述规范和数据标准. 北京: 中国农业出版社, 2006.
	WANG X Y, WANG L. Descriptors and Data Standard for Broomcorn Millet (Panicum miliaceum L.). Beijing: China Agriculture Press, 2006. (in Chinese)
[2]	CRAWFORD G W. Agricultural origins in North China pushed back to the Pleistocene-Holocene boundary. Proceedings of the National Academy of Sciences of the United States of America, 2009, 106(18): 7271-7272.
[3]	LU H Y, ZHANG J P, LIU K B, WU N Q, LI Y M, ZHOU K S, YE M L, ZHANG T Y, ZHANG H J, YANG X Y, SHEN L C, XU D K, LI Q. Earliest domestication of common millet (Panicum miliaceum) in East Asia extended to 10000 years ago. Proceedings of the National Academy of Sciences of the United States of America, 2009, 106(18): 7367-7372.
[4]	MARTIN J H, LEONARD W H, STAMP D L. Principles of Field Crop Production. 3rd ed. New York: MacMillan Press, 1976: 415-429.
[5]	乔治军. 糜子产业发展现状与思路. 作物杂志, 2013(5): 25-27.
	QIAO Z J. Present situation and developing thought of broomcorn millet industry. Crops, 2013(5): 25-27. (in Chinese)
[6]	柴岩, 冯佰利, 王宏岩. 中国黄米食品. 杨凌: 西北农林科技大学出版社, 2012.
	CHAI Y, FENG B L, WANG H Y. Food of Proso Millet in China. Yangling: Northwest A & Forestry University Press, 2012. (in Chinese)
[7]	王瑞云, 杨阳, 王海岗, 陈凌, 王纶, 陆平, 刘敏轩, 乔治军. 糜子PmNCED1的克隆及其对PEG胁迫的响应. 核农学报, 2018, 32(2): 244-256. doi: 10.11869/j.issn.100-8551.2018.02.0244
	WANG R Y, YANG Y, WANG H G, CHEN L, WANG L, LU P, LIU M X, QIAO Z J. Cloning of gene PmNCED1 and its response to PEG stress in common millet. Journal of Nuclear Agricultural Sciences, 2018, 32(2): 244-256. (in Chinese)
[8]	王瑞云. 糜子遗传多样性及进化研究进展. 北京: 中国农业出版社, 2017.
	WANG R Y. Genetic Diversity and Evolution Advancement in Common Millet (Panicum miliaceum L.). Beijing: China Agriculture Press, 2017. (in Chinese)
[9]	王纶, 王星玉. 中国黍稷种质资源研究. 北京: 中国农业科学技术出版社, 2018.
	WANG L, WANG X Y. Study on Common Millet Germplasm Resources in China. Beijing: China Agricultural Science and Technology Press, 2018. (in Chinese)
[10]	陈小红, 何杰丽, 石甜甜, 邵欢欢, 王海岗, 陈凌, 高志军, 王瑞云, 乔治军. 基于转录组测序开发糜子SSR标记. 中国农业科学, 2020, 53(10): 1940-1949. doi: 10.3864/j.issn.0578-1752.2020.10.002
	CHEN X H, HE J L, SHI T T, SHAO H H, WANG H G, CHEN L, GAO Z J, WANG R Y, QIAO Z J. Developing SSR markers of proso millet based on transcriptome sequencing. Scientia Agricultura Sinica, 2020, 53(10): 1940-1949. (in Chinese) doi: 10.3864/j.issn.0578-1752.2020.10.002
[11]	薛延桃, 陆平, 乔治军, 刘敏轩, 王瑞云. 基于SSR标记的黍稷种质资源遗传多样性及亲缘关系研究. 中国农业科学, 2018, 51(15): 2846-2859. doi: 10.3864/j.issn.0578-1752.2018.15.002
	XUE Y T, LU P, QIAO Z J, LIU M X, WANG R Y. Genetic diversity and genetic relationship of broomcorn millet (Panicum miliaceum L.) germplasm based on SSR markers. Scientia Agricultura Sinica, 2018, 51(15): 2846-2859. (in Chinese)
[12]	石甜甜, 何杰丽, 高志军, 陈凌, 王海岗, 乔治军, 王瑞云. 利用EST-SSR评估糜子资源遗传差异. 中国农业科学, 2019, 52(22): 4100-4109. doi: 10.3864/j.issn.0578-1752.2019.22.014
	SHI T T, HE J L, GAO Z J, CHEN L, WANG H G, QIAO Z J, WANG R Y. Genetic diversity of common millet resources assessed with EST-SSR markers. Scientia Agricultura Sinica, 2019, 52(22): 4100-4109. (in Chinese) doi: 10.3864/j.issn.0578-1752.2019.22.014
[13]	连帅, 陆平, 乔治军, 张琦, 张茜, 刘敏轩, 王瑞云. 利用SSR分子标记研究国内外黍稷地方品种和野生资源的遗传多样性. 中国农业科学, 2016, 49(17): 3264-3275. doi: 10.3864/j.issn.0578-1752.2016.17.002
	LIAN S, LU P, QIAO Z J, ZHANG Q, ZHANG Q, LIU M X, WANG R Y. Genetic diversity in broomcorn millet (Panicum miliaceum L.) from China and abroad by using SSR markers. Scientia Agricultura Sinica, 2016, 49(17): 3264-3275. (in Chinese)
[14]	TIAN H L, WANG F G, ZHAO J R, YI H M, WANG L, WANG R, YANG Y, SONG W. Development of maize SNP3072, a high- throughput compatible SNP array, for DNA fingerprinting identification of Chinese maize varieties. Molecular Breeding, 2015, 35(6): 136. doi: 10.1007/s11032-015-0335-0
[15]	王凤格, 杨扬, 易红梅, 赵久然, 任洁, 王璐, 葛建镕, 江彬, 张宪晨, 田红丽, 侯振华. 中国玉米审定品种标准SSR指纹库的构建. 中国农业科学, 2017, 50(1): 1-14.
	WANG F G, YANG Y, YI H M, ZHAO J R, REN J, WANG L, GE J R, JIANG B, ZHANG X C, TIAN H L, HOU Z H. Construction of an SSR-based standard fingerprint database for corn variety authorized in China. Scientia Agricultura Sinica, 2017, 50(1): 1-14. (in Chinese)
[16]	赵艳杰, 冯艳芳, 黄思思, 马莹雪, 李嫒嫒, 张蝶, 邓超, 韩瑞玺, 唐浩. 182份东北地区受保护大豆品种DNA指纹库的构建及分析. 中国种业, 2019(11): 43-47.
	ZHAO Y J, FENG Y F, HUANG S S, MA Y X, LI A A, ZHANG D, DENG C, HAN R X, TANG H. Construction and analysis of DNA fingerprint database of 182 protected soybean varieties in Northeast China. China Seed Industry, 2019(11): 43-47. (in Chinese)
[17]	李清, 罗永坚, 吴柔贤, 贾俊婷, 张文虎, 宋松泉, 刘军. 广东省大豆种质资源遗传多样性分析及DNA分子身份证构建. 广东农业科学, 2020, 47(12): 221-228.
	LI Q, LUO Y J, WU R X, JIA J T, ZHANG W H, SONG S Q, LIU J. Analysis on genetic diversity and construction of DNA molecular identity card of soybean germplasm resources in Guangdong Province. Guangdong Agricultural Sciences, 2020, 47(12): 221-228. (in Chinese)
[18]	樊晓静, 于文涛, 蔡春平, 林浥, 王泽涵, 房婉萍, 张见明, 叶乃兴. 利用SNP标记构建茶树品种资源分子身份证. 中国农业科学, 2021, 54(8): 1751-1760. doi: 10.3864/j.issn.0578-1752.2021.08.014
	FAN X J, YU W T, CAI C P, LIN Y, WANG Z H, FANG W P, ZHANG J M, YE N X. Construction of molecular ID for tea cultivars by using of single nucleotide polymorphism (SNP) markers. Scientia Agricultura Sinica, 2021, 54(8): 1751-1760. (in Chinese)
[19]	张枭, 杜潇, 孙馨宇, 王键, 董文轩. 利用SSR标记构建部分山楂资源的基因身份证. 沈阳农业大学学报, 2021, 52(2): 153-159.
	ZHANG X, DU X, SUN X Y, WANG J, DONG W X. Construction of molecular identity for partial Crataegus resources based on SSR markers. Journal of Shenyang Agricultural University, 2021, 52(2): 153-159. (in Chinese)
[20]	刘晓鑫, 谢传晓, 赵琦, 路明, 曲延英, 张滨, 梁业红, 张世煌. 基于SSR荧光标记技术的玉米群体混合样本基因频率分析方法. 中国农业科学, 2008, 41(12): 3991-3998.
	LIU X X, XIE C X, ZHAO Q, LU M, QU Y Y, ZHANG B, LIANG Y H, ZHANG S H. Establishment of fluorescent SSR technique on detecting allelic frequency in maize (Zea mays L.) populations with bulk sampling strategy. Scientia Agricultura Sinica, 2008, 41(12): 3991-3998. (in Chinese)
[21]	王瑞云, 季煦, 陆平, 刘敏轩, 许月, 王纶, 王海岗, 乔治军. 利用荧光SSR分析中国糜子遗传多样性. 作物学报, 2017, 43(4): 530-548.
	WANG R Y, JI X, LU P, LIU M X, XU Y, WANG L, WANG H G, QIAO Z J. Analysis of genetic diversity in common millet (Panicum miliaceum L.) using fluorescent SSR in China. Acta Agronomica Sinica, 2017, 43(4): 530-548. (in Chinese) doi: 10.3724/SP.J.1006.2017.00530
[22]	冉昆, 隋静, 王宏伟, 魏树伟, 张勇, 董冉, 董肖昌, 王少敏. 利用SSR荧光标记构建山东地方梨种质资源分子身份证. 果树学报, 2018, 35(S1): 71-78.
	RAN K, SUI J, WANG H W, WEI S W, ZHANG Y, DONG R, DONG X C, WANG S M. Using the fluorescent labeled SSR markers to establish the molecular ID of pear germplasm resources in Shandong. Journal of Fruit Science, 2018, 35(S1): 71-78. (in Chinese)
[23]	郭艳春, 张力岚, 陈思远, 祁建民, 方平平, 陶爱芬, 张列梅, 张立武. 黄麻应用核心种质的DNA分子身份证构建. 作物学报, 2021, 47(1): 80-93. doi: 10.3724/SP.J.1006.2021.04022
	GUO Y C, ZHANG L L, CHEN S Y, QI J M, FANG P P, TAO A F, ZHANG L M, ZHANG L W. Establishment of DNA molecular fingerprint of applied core germplasm in jute (Corchorus spp.). Acta Agronomica Sinica, 2021, 47(1): 80-93. (in Chinese) doi: 10.3724/SP.J.1006.2021.04022
[24]	郑永胜, 张晗, 王东建, 孙加梅, 王雪梅, 段丽丽, 李华, 王玮, 李汝玉. 基于荧光检测技术的小麦品种SSR鉴定体系的建立. 中国农业科学, 2014, 47(19): 3725-3735. doi: 10.3864/j.issn.0578-1752.2014.19.001
	ZHENG Y S, ZHANG H, WANG D J, SUN J M, WANG X M, DUAN L L, LI H, WANG W, LI R Y. Development of a wheat variety identification system based on fluorescently labeled SSR markers. Scientia Agricultura Sinica, 2014, 47(19): 3725-3735. (in Chinese)
[25]	连帅. 黍稷种质资源遗传多样性研究及微核心种质的构建[D]. 太谷: 山西农业大学, 2017.
	LIAN S. Genetic diversity assessment of broomcorn millet (Panicum miliaceum L.) and construction of a mini core collection in China[D]. Taigu: Shanxi Agricultural University, 2022. (in Chinese)
[26]	EDWARDS K, JOHNSTONE C, THOMPSON C. A simple and rapid method for the preparation of plant genomic DNA for PCR analysis. Nucleic Acids Research, 1991, 19(6): 1349. doi: 10.1093/nar/19.6.1349 pmid: 2030957
[27]	王瑞云, 刘笑瑜, 王海岗, 陆平, 刘敏轩, 陈凌, 乔治军. 用高基元微卫星标记分析中国糜子遗传多样性. 中国农业科学, 2017, 50(20): 3848-3859. doi: 10.3864/j.issn.0578-1752.2017.20.002
	WANG R Y, LIU X Y, WANG H G, LU P, LIU M X, CHEN L, QIAO Z J. Evaluation of genetic diversity of common millet (Panicum miliaceum L.) germplasm available in China using high motif nucleotide repeat SSR markers. Scientia Agricultura Sinica, 2017, 50(20): 3848-3859. (in Chinese)
[28]	何杰丽, 石甜甜, 陈凌, 王海岗, 高志军, 杨美红, 王瑞云, 乔治军. 糜子EST-SSR分子标记的开发及种质资源遗传多样性分析. 植物学报, 2019, 54(6): 723-732. doi: 10.11983/CBB19037
	HE J L, SHI T T, CHEN L, WANG H G, GAO Z J, YANG M H, WANG R Y, QIAO Z J. The genetic diversity of common millet (Panicum miliaceum L.) germplasm resources based on the EST-SSR markers. Chinese Bulletin of Botany, 2019, 54(6): 723-732. (in Chinese)
[29]	薛亚鹏, 李元阳, 陈凌, 王海岗, 王瑞云, 乔治军. 山西省糜子DNA分子身份证的构建. 山西农业大学学报(自然科学版), 2022, 42(1): 19-25.
	XUE Y P, LI Y Y, CHEN L, WANG H G, WANG R Y, QIAO Z J. Construction of DNA molecular identity card of broomcorn millet germplasm in Shanxi Province. Journal of Shanxi Agricultural University (Natural Science Edition), 2022, 42(1): 19-25. (in Chinese)
[30]	LIU K J, MUSE S V. PowerMarker: An integrated analysis environment for genetic marker analysis. Bioinformatics, 2005, 21(9): 2128-2129. doi: 10.1093/bioinformatics/bti282 pmid: 15705655
[31]	YEH F C. Population genetic analysis of co-dominant and dominant markers and quantitative traits. Belgian Journal of Botany, 1997, 129: 157.
[32]	TAMURA K, PETERSON D, PETERSON N, STECHER G, NEI M, KUMAR S. MEGA5: Molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Molecular Biology and Evolution, 2011, 28(10): 2731-2739. doi: 10.1093/molbev/msr121 pmid: 21546353
[33]	寇淑君, 霍阿红, 付国庆, 纪军建, 王瑶, 左振兴, 刘敏轩, 陆平. 利用荧光SSR分析中国糜子的遗传多样性和群体遗传结构. 中国农业科学, 2019, 52(9): 1475-1487. doi: 10.3864/j.issn.0578-1752.2019.09.001
	KOU S J, HUO A H, FU G Q, JI J J, WANG Y, ZUO Z X, LIU M X, LU P. Genetic diversity and population structure of broomcorn millet in China based on fluorescently labeled SSR. . Scientia Agricultura Sinica, 2019, 52(9): 1475-1487. (in Chinese) doi: 10.3864/j.issn.0578-1752.2019.09.001
[34]	SUN Y L, LIU Y, SHI J F, WANG L, LIANG C Z, YANG J, CHEN J F, CHEN M S. Biased mutations and gene losses underlying diploidization of the tetraploid broomcorn millet genome. The Plant Journal, 2023, 113, 787-801. doi: 10.1111/tpj.v113.4
[35]	丁敏, 段政勇, 王宇卓, 薛亚鹏, 王海岗, 陈凌, 王瑞云, 乔治军. 糜子GBSSI基因功能标记的开发与验证. 作物学报, 2023, 49(3): 703-718. doi: 10.3724/SP.J.1006.2023.24028
	DING M, DUAN Z Y, WANG Y Z, XUE Y P, WANG H G, CHEN L, WANG R Y, QIAO Z J. Development and validation of functional markers of GBSSI gene in proso millet. Acta Agronomica Sinica, 2023, 49(3): 703-718. (in Chinese) doi: 10.3724/SP.J.1006.2023.24028
[36]	沈群. 杂粮与科学的美味邂逅. 北京: 中国农业大学出版社, 2022.
	SHEN Q. Delicious Encounters Between Grains and Science. Beijing: Chinese Agricultural University Press, 2022. (in Chinese)
[37]	郝晨阳, 王兰芬, 贾继增, 董玉琛, 张学勇. SSR荧光标记和银染技术的比较分析. 作物学报, 2005, 31(2): 144-149.
	HAO C Y, WANG L F, JIA J Z, DONG Y C, ZHANG X Y. Comparison of fluorescence and silver-staining detection systems of microsatellite markers. Acta Agronomica Sinica, 2005, 31(2): 144-149. (in Chinese)
[38]	陈小红, 林元香, 王倩, 丁敏, 王海岗, 陈凌, 高志军, 王瑞云, 乔治军. 基于高基元SSR构建黍稷种质资源的分子身份证. 作物学报, 2022, 48(4): 908-919. doi: 10.3724/SP.J.1006.2022.14034
	CHEN X H, LIN Y X, WANG Q, DING M, WANG H G, CHEN L, GAO Z J, WANG R Y, QIAO Z J. Development of DNA molecular ID card in hog millet germplasm based on high motif SSR. Acta Agronomica Sinica, 2022, 48(4): 908-919. (in Chinese) doi: 10.3724/SP.J.1006.2022.14034
[39]	林元香. 山西糜子核心种质分子身份证的构建[D]. 太谷: 山西农业大学, 2022.
	LIN Y X. Construction of molecular ID card of core germplasm of common millet (Panicum miliaceum L.) in Shanxi[D]. Taigu: Shanxi Agricultural University, 2022. (in Chinese)

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材料编号 Serial number	统一编号 Unicode	名称 Name	来源 Source	备注 Remark
1	00000082	黄糜子Huangmeizi	中国黑龙江Heilongjiang, China	地方品种Landrace
2	00000840	六十天小红黍60-day Xiaohongshu	中国山西Shanxi, China	地方品种Landrace
3	00000710	皋兰鸭蛋青Gaolanyadanqing	中国甘肃Gansu, China	地方品种Landrace
4	00003117	黄糜Huangmei	中国新疆Xinjiang, China	地方品种Landrace
5	00006438	乌克兰黍Ukraine shu	中国内蒙古Inner Mongolia, China	地方品种Landrace
6	00004198	金守黍Jinshoushu	中国广东Guangdong, China	地方品种Landrace
7	00007131	白黍子Baishuzi	中国河北Hebei, China	地方品种Landrace
8	00006681	札达糜Zhadamei	中国西藏Tibet, China	地方品种Landrace
9	00007258	黄糜子Huangmeizi	中国辽宁Liaoning, China	地方品种Landrace
10	00007375	790035	印度India	地方品种Landrace
11	00005191	塞盖德斯Saigaidesi	中国山东Shandong, China	地方品种Landrace
12	00005671	扶余白糜子Fuyubaimeizi	中国吉林Jilin, China	地方品种Landrace
13	00005452	夯糜子Hangmeizi	中国陕西Shaanxi, China	地方品种Landrace
14	00002641	西吉小黄糜Xijixiaohuangmei	中国宁夏Ningxia, China	地方品种Landrace

位点 Locus	正向引物 Forward primer (5′-3′)	反向引物 Reverse primer (5′-3′)	5′端修饰 5′ end modification	重复序列 Repeated motif	退火温度 Tm (℃)
RYW3	GGAGGCGTGACAATAAAAC	GGCGTGAGGTGTTGTTTTT	5′6-FAM	(CTGCAA)₅	48.90
RYW5	GACGATGCTCTTGACCTTGT	CACCGTGAAATGTCTCTGCT	5′6-FAM	(CCTTT)₅	51.80
RYW6	AGCCGATTTGCTGTGGAGT	CTGCCTCCGATGAGTTGGT	5′6-FAM	(ACACC)₅	52.15
RYW11	TGCTCGTCTTCTCGCTTCG	AGTAGTCCTCCACCGCCATCT	5′6-FAM	(GGTA)₅	54.75
RYW14	CGCACAACGACCACAAGAG	ATACACCAGAGGAGCACGC	5′6-FAM	(GGCC)₅	57.35
RYW18	CTCCCTCTTTGTCCTCGTT	GCTGCCTCTTCGCTATCTT	5′6-FAM	(AGTT)₆	51.10
RYW20	ACCTCTTGCCGCACACTAC	TTCTACATCCCCGAACCAC	5′6-FAM	(TTGG)₆	52.15
RYW35	ATTAGCATCCCCCTCCAC	ATCCGCTTTCCCAACCAC	5′6-FAM	(CGTGC)₅(GGA)₆	50.30
RYW37	CATTCCGTTCCTTGTCTTCC	CAGTCTCACTCCTGCGATGT	5′6-FAM	(GCGAT)₅	52.80
RYW40	TGCTCTTCGGCTCTTCTCC	ATCAGCTCATCGTGACCCC	5′6-FAM	(CAGC)₆	53.20
RYW43	GGAGATGCTTGCTTGGTTG	CAGGAATCGCAAGGAACAG	5′HEX	(GGAG)₅	54.00
RYW54	GCACTTGCTCCTGCTTCTC	GACCTTGCCGATGTTGTTG	5′HEX	(CCTC)₅	55.65
RYW124	CCACAAAGCGAAAGAACC	GAGTCCAAGCCCTCATCC	5′6-FAM	(TCG)₆	53.65
RYW125	TTGACGACGACTGTGTGC	TGTTGGTGGAGTTGAGGAC	5′6-FAM	(GGC)₅	55.10
RYW146	TGATGCTTCTTGGGTTCG	CGCCGTCCACTTCTGTAT	5′6-FAM	(GCG)₆	53.60

引物 Primer	观测等位变异 Observed number of alleles (Na)	Shannon 多样性指数 Shannon’s diversity index(I)	观测杂合度 Observed heterozygosity (Ho)	期望杂合度 Expected heterozygosity (He)	Nei’s 基因多样性指数 Nei's expected heterozygosity (Nei)	多态性信息含量Polymorphism information content (PIC)
RYW3	15.0000	0.9874	0.8136	0.4114	0.4105	0.5073
RYW6	25.0000	2.0587	0.5238	0.7469	0.7452	0.8071
RYW11	3.0000	0.7178	0.0086	0.5053	0.5043	0.0334
RYW18	4.0000	0.2055	0.9455	0.0795	0.0793	0.2103
RYW37	4.0000	1.1777	0.3376	0.6700	0.6686	0.6495
RYW125	16.0000	1.6084	0.4414	0.6940	0.6916	0.7681
RYW43	20.0000	1.2230	0.5399	0.5161	0.5149	0.6540
合计Total	87.0000	-	-	-	-	-
平均Mean	12.4286	1.1398	0.5158	0.5176	0.5163	0.5185

引物/组合 Primer/Combination	区分种质数 No. of germplasm distinguished	区分率 Rate of discrimination (%)
RYW3	10	4.26
RYW6	17	7.23
RYW11	2	0.85
RYW18	0	0.00
RYW37	2	0.85
RYW43	16	6.81
RYW125	16	6.81
RYW3+RYW6	46	19.57
RYW3+RYW6+RYW11	48	20.42
RYW3+RYW6+RYW11+RYW18	62	26.38
RYW3+RYW6+RYW11+RYW18+RYW37	88	37.45
RYW3+RYW6+RYW11+RYW18+RYW37+RYW43	161	68.51
RYW3+RYW6+RYW11+RYW18+RYW37+RYW43+RYW125	235	100.00

Construction of DNA Molecular Identity Card of Core Germplasm of Broomcorn Millet in China Based on Fluorescence SSR

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