基于SSR标记的雪茄烟种质资源指纹图谱库的构建及遗传多样性分析

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王琰琰¹^,^**, 王俊²^,^**, 刘国祥¹, 钟秋², 张华述², 骆铮珍², 陈志华³, 戴培刚¹, 佟英¹, 李媛¹, 蒋勋¹, 张兴伟^,¹^,^*, 杨爱国^,¹^,^*

¹中国农业科学院烟草研究所, 山东青岛 266101

²四川省烟草公司德阳市公司, 四川德阳 618400

³四川省烟草科学研究所, 四川成都 610000

Construction of SSR fingerprint database and genetic diversity analysis of cigar germplasm resources

WANG Yan-Yan¹^,^**, WANG Jun²^,^**, LIU Guo-Xiang¹, ZHONG Qiu², ZHANG Hua-Shu², LUO Zheng-Zhen², CHEN Zhi-Hua³, DAI Pei-Gang¹, TONG Ying¹, LI Yuan¹, JIANG Xun¹, ZHANG Xing-Wei^,¹^,^*, YANG Ai-Guo^,¹^,^*

¹Tobacco Research Institute, Chinese Academy of Agricultural Sciences, Qingdao 266101, Shandong, China

²Deyang Tobacco Company of Sichuan Province, Deyang 618400, Sichuan, China

³Sichuan Institute of Tobacco Science, Chengdu 610000, Sichuan, China

通讯作者: ^*张兴伟, E-mail: zhangxingwei@caas.cn; ^*杨爱国, E-mail: yangaiguo@caas.cn

^** 同等贡献(Contributed equally to this work)
收稿日期:2020-08-11接受日期:2020-12-1网络出版日期:2021-01-12

基金资助:

本研究由中国烟草总公司四川省公司科技项目.SCYC201903
中国农业科学院科技创新工程.ASTIP-TRIC01
中央级公益性科研院所基本科研业务费专项.1610232019008
农作物种质资源保护与利用专项.2020NWB038
国家作物种质资源库资助.NCGRC-2020-18

Received:2020-08-11Accepted:2020-12-1Online:2021-01-12

Fund supported:

This study was supported by the Science and Technology Project of China National Tobacco Corporation Sichuan Branch.SCYC201903
the Science and Technology Innovation Project of Chinese Academy of Agricultural Sciences .ASTIP-TRIC01
the Special Funds for Fundamental Scientific Research Operation of Central-level Public Welfare Scientific Research Institutes.1610232019008
the Special Project for Protection and Utilization of Crop Germplasm Resources.2020NWB038
the National Crop Germplasm Bank.NCGRC-2020-18

作者简介 About authors
王琰琰, E-mail: wangyy101@163.com;

王俊, E-mail: 599187325@qq.com

摘要
为从分子水平研究我国雪茄烟种质资源的遗传多样性差异并建立雪茄烟品种的DNA指纹图谱数据库, 本研究利用43对多态性好的SSR引物对220份雪茄烟种质进行遗传多样性分析, 筛选出14对核心引物对雪茄烟种质进行指纹图谱的构建。结果表明, 43对SSR引物在220份雪茄烟种质材料中共扩增出243个等位基因, 平均每个标记5.65个, 变幅为2~13, 每个位点的多态性信息量(polymorphism information content, PIC)变化为0.2078~0.9087, 平均为0.6360。有效等位基因数(number of effective alleles, N_e)范围为1.3081~11.7876, 平均有效等位基因数为3.9077; 观测杂合度(observed heterozygosity, H_o)变化范围为0.0828~0.7639, 平均为0.3191; 预期杂合度(expected heterozygosity, H_e)的变化范围为0.2361~0.9172, 平均为0.6809; 种群平均Shannon遗传多样性指数(Shannon genetic diversity index, I)为1.3756, 遗传距离在0.0233~0.9286之间, 平均遗传距离0.6816。聚类分析表明, 在遗传距离为0.74处, 可将供试雪茄烟资源分为3个类群。Structure群体遗传结构分析和主成分分析将所有的供试材料划分为2个类群。根据引物的分析和表型鉴定结果, 确定良种、辅善和满耳朵, 山东大叶和牡丹江05-1, Florida 513和CA0701为异名同种, 一个品种保留一份种质, 剩余216份不同种质。从43对SSR引物中筛选出14对可区分所有供试材料的SSR引物作为核心引物构建了216个雪茄烟品种的指纹图谱。我国雪茄烟种质资源具有较高水平的遗传多样性, 本研究构建的雪茄烟种质资源SSR指纹图谱库及遗传分析的结果在分子水平上为筛选、鉴定优质雪茄烟种质资源、挖掘重要基因以及拓宽雪茄烟遗传育种基础等工作提供科学依据。
关键词： 雪茄烟;种质资源;SSR标记;遗传多样性;指纹图谱

Abstract

In order to investigate the genetic diversity of Chinese cigar germplasm resources in molecular level and to establish the DNA fingerprint database of cigar varieties, 43 pairs of SSR primers with good polymorphic were used to analyze the genetic diversity of 220 cigar germplasms, and 14 pairs of core primers were screened from 43 pairs of SSR primers to construct the fingerprint database of 220 cigar germplasms in the study. Results showed that a total of 243 alleles were amplified by 43 pairs of SSR primers in 220 cigar germplasm materials, with an average of 5.65 per marker, ranging from 2 to 13. The polymorphic information content (PIC) of each locus varied from 0.2078 to 0.9087 with an average of 0.6360. The number of effective alleles (N_e) ranged from 1.3081 to 11.7876, and the average number of effective alleles was 3.9077. The observed heterozygosity (H_o) ranges from 0.0828 to 0.7639 with an average of 0.3191. The expected heterozygosity (H_e) ranged from 0.2361 to 0.9172 with an average of 0.6809. The average Shannon genetic diversity index (I) was 1.3756, the genetic distance was between 0.0233 and 0.9286, and the average genetic distance was 0.6816. Unweighted pair-group method with arithmetic means (UPGMA) cluster analysis showed that the tested cigar resources can be divided into three groups at the genetic distance of 0.74. All tested materials were divided into two groups by population genetic structure analysis and principal component analysis (PCA). Based on the primer analysis and phenotypic identification, it was determined that the ‘LZ’, ‘FS’ and ‘MED’, ‘SDDY’ and ‘MDJ 05-1’, ‘Florida 513’ and ‘CA0701’ were the same species with different names. One variety retained one germplasm, leaving 216 different germplasm. 14 pairs of SSR primers which could distinguish all the tested materials were screened to construct the fingerprint database of 216 cigar cultivars from 43 pairs of SSR primers as core primers. In conclusion, cigar germplasm resources in China had a high level of genetic diversity. In this study, the constructed SSR fingerprint database and genetic analysis of cigar germplasm resources provided the scientific basis for the screening and identification of high-quality cigar germplasms resources, the mining of important genes, and the broadening the basis of cigar genetic breeding.

Keywords：cigar;germplasm resources;SSR marker;genetic diversity;fingerprint database

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本文引用格式
王琰琰, 王俊, 刘国祥, 钟秋, 张华述, 骆铮珍, 陈志华, 戴培刚, 佟英, 李媛, 蒋勋, 张兴伟, 杨爱国. 基于SSR标记的雪茄烟种质资源指纹图谱库的构建及遗传多样性分析[J]. 作物学报, 2021, 47(7): 1259-1274. doi:10.3724/SP.J.1006.2021.04183
WANG Yan-Yan, WANG Jun, LIU Guo-Xiang, ZHONG Qiu, ZHANG Hua-Shu, LUO Zheng-Zhen, CHEN Zhi-Hua, DAI Pei-Gang, TONG Ying, LI Yuan, JIANG Xun, ZHANG Xing-Wei, YANG Ai-Guo. Construction of SSR fingerprint database and genetic diversity analysis of cigar germplasm resources[J]. Acta Crops Sinica, 2021, 47(7): 1259-1274. doi:10.3724/SP.J.1006.2021.04183

烟草(Nicotiana tabacum L.)为双子叶植物纲(Dicotyledoneae)管花目(Tubiflorae)茄科(Solanaceae)烟属(Nicotiana)一年生或有限多年生草本植物^[1], 种植范围十分广泛, 是我国重要的经济作物之一, 也是最早应用于分子生物学和基因工程研究的模式植物之一^[2]。我国烟草种质资源十分丰富。目前已有76个烟属植物种被发现, 其中可直接利用的栽培种有普通烟草(N. tabacum L.)和黄花烟草(N. rustica L.)^[3,4,5]。雪茄烟是普通烟草的一种, 我国雪茄烟种质资源数量比较匮乏, 其仅占烟草种质资源的3% ^[6], 且主要为国外引进种质。由于我国雪茄烟种质资源引种渠道多元, 保存较为分散, 种质保存机构间相互引种且独立命名, 因此存在种质重复引进、品种同种异名、同名异种, 以及种质间亲缘关系不清等问题, 给烟草种质资源编目保存、育种亲本选择及优良品种推广应用带来不便。因此, 应用分子标记技术分析不同雪茄烟品种间的亲缘关系和遗传差异, 探索其遗传多样性的丰富程度, 并在此基础上构建雪茄烟种质资源指纹图谱库, 对于我国雪茄烟资源品种鉴定、品种选育及资源高效利用具有重要意义。

目前, 常用的分子标记有随机引物多态性标记(random amplified polymorphic DNA, RAPD)^[7,8,9]、扩增片段长度多态性标记(amplified fragment length polymorphism, AFLP)^[10,11,12]、简单重复序列(simple sequence repeats, SSR)、相关序列扩增多态性(sequence-related amplified polymorphism, SRAP)^[13,14]、核糖体DNA和简单序列重复(inter-simple sequence repeat, ISSR)^[15,16,17]、单核苷酸多态性(single nucleotide polymorphism, SNP)^[18,19]等。在众多分子标记手段中, SSR分子标记由于具有灵敏度高、多态性好、稳定可靠等优点, 是分析遗传多样性及群体亲缘关系很好的标记方法。SSR标记技术已经被广泛用于作物品种鉴定^[20,21]、多样性分析^[22,23,24]、指纹图谱的构建^[25,26]等研究中。近些年在烟草中也得到了较多应用, Bindler等^[27]在2007年公布了第1张烟草SSR标记遗传连锁图谱, 并于2011年在原有的工作基础上又构建了包括2317个标记, 2363个位点的高密度遗传图谱^[28], 为SSR分子标记的广泛应用奠定基础; Moon等^[29]在2008年利用46对SSR引物分析了3种类型的54份烟草属材料的遗传多样性, 于2009年利用71对SSR引物对117个美国主栽烟草品种进行遗传多样性分析^[30], 同年用70个SSR标记对来自美国烟草种质资源收集的702个材料进行了遗传多样性分析^[31]; 马冰等^[32]利用24对核心引物对20份烤烟种质进行了遗传多样性分析; 张雪廷等^[33]利用30对SSR引物分析了38份晾晒烟种质资源的遗传关系; 童治军等^[34]利用120个SSR标记分析了231份烤烟种质资源的遗传多样性及群体遗传结构; 刘国祥等^[35]利用25对SSR标记对33份晒烟材料进行了遗传多样性分析并构建了DNA指纹图谱; 张铭真等^[36]利用64对引物分析了81份烟草种质资源的遗传多样性; 陈芳等^[37]利用48对SSR标记对80份烟草种质资源进行遗传多样性分析, 并绘制了SSR计算机数字指纹图谱; 目前为止国内外报道的有关烟草方面遗传多样性研究多为烤烟和晒晾烟, 而利用SSR标记对雪茄烟种质资源遗传关系进行分析并构建DNA指纹图谱, 在国内外尚未见报道。

本研究利用SSR分子标记对供试雪茄烟种质资源进行遗传多样性分析和指纹图谱的绘制, 以期确定烟草种质材料间的遗传背景差异及品种间亲缘关系, 为雪茄烟种质资源的鉴定、研究和利用提供依据, 并为后续充分发掘利用雪茄烟种质资源、选配合适的育种亲本及种质资源收集、鉴定、编目、保存等育种工作奠定基础。

1 材料与方法

1.1 试验材料

试验所采用的220份雪茄烟种质材料(附表1)由国家烟草种质资源中期库(中国农业科学院烟草研究所)、德阳市烟草公司提供。表1中的晒烟、晾烟、白肋烟及烤烟等类型材料均为选育或改良后的具有雪茄型香气的品种, 经多年试验已投入到雪茄烟选育及生产中, 因此这里统一称为雪茄烟。

1.2 基因组DNA的提取

供试材料于2019年种植在四川省什邡市师古镇苏家桥试验基地。取苗期幼嫩叶片于2 mL离心管中, 并快速放入干冰中冷冻, 带回实验室后置于-80℃的低温冰箱中保存。置于磨样机磨碎后利用SLS法提取DNA, 加ddH₂O在4℃条件下溶解12 h。用核酸测定仪(Thermo Scientific NanoProp 2000)测定DNA浓度, 将DNA浓度统一稀释到50 ng μL^-1, -20℃低温下保存。

1.3 SSR引物

试验所用烟草SSR标记引物是从Bindler等^[28]公布的2317对SSR引物中经初步筛选得到的2000对扩增效果好的SSR引物, 由六合华大(北京)基因科技有限公司合成。

1.4 PCR扩增

扩增体系为10 μL, 包括1 μL的DNA (浓度50 ng μL^-1)、2×Rapid Taq Master Mix (含Taq DNA聚合酶、dNTPs、MgCl₂以及反应缓冲液) 5 μL、上游引物1 μL、下游引物1 μL, 剩余体系用ddH₂O定量。总反应程序为95℃预变性5 min; 95℃变性15 s, 58℃ (根据T_m值进行调整)退火15 s, 72℃延伸15 s, 循环35次; 72℃延伸8 min。PCR扩增产物在6%的非变性聚丙烯酰胺凝胶上电泳分离, 上样量为1.0 μL, 电压1200 V, 功率60 W下电泳1.5 h后银染显色, 照相保存。

1.5 数据分析

读取条带信息时, 有条带的标记为“1”, 无条带的记为“0”, 条带缺失的记为“9”, 构建数列矩阵, 用DataFormater软件^[38]将读取的数据进行矩阵转换。用Popgene32软件^[39]计算观察等位基因数(N_o)、有效等位基因数(N_e)、Nei’s基因多样性指数(H)、Shannon’s多态信息指数(I)。并用PIC_CALC Version 0.6软件^[40]计算多态信息量(PIC):

, 其中n为该位点的等位基因数, P_i为第i个等位基因在群体中的频率, P_j为第j个等位基因在群体中的频率。材料间遗传相似系数(genetic similarity coefficient, GS)和遗传距离(genetic distance, GD)按照公式: GS = m/(m+n), GD = 1-GS来计算, 其中, m为基因型间共有带数目, n为差异带数目。利用MEGA7.0.26软件^[41]以非加权配对法(unweighted pair group method with arithmetic mean, UPGMA)进行聚类分析并绘制出聚类图。利用NTSYS-PC 2.10软件中DCENTER和EIGEN模块对220份雪茄烟资源间进行主成分分析(principal component analysis, PCA)。利用Structure 2.3.4软件分析群体遗传结构, 估计最佳群体组群数K的取值范围为设置在1~15, 将参数burn-in period设为10,000, MCMC设为100,000^[42], 每个K值重复运行5次, 依据似然值最大原则选取合适的K值为群体数目, 当K值持续增大时, 参考Evanno等^[43]的方法计算ΔK来选择合适的K值并计算出Q值。

1.6 指纹图谱构建

根据利用尽量少的引物区分尽量多的品种的原则, 首先选取能够把供试材料完全区分开的部分引物作为核心引物构建供试材料指纹图谱代码, 然后参考王静毅等^[44]和宋海斌等^[45]的图谱代码构建方法进行构建。把每对核心SSR引物扩增出的条带, 按其分子量从小到大的顺序按1、2、3……进行编号, 记录各引物对材料扩增出的条带编号, 没有扩增出条带的用0表示。将核心引物按照顺序用A、B、C……进行编号, 每个品种的DNA经不同的引物扩增后将会形成由字母和阿拉伯数字组成的一系列带型编号, 便形成了该品种的SSR指纹图谱代码。利用在线软件草料二维码生成器(http://cli.im/)对每个品种进行编码, 将各个品种的品种名称、全国统一编号、类型、植物学分类、指纹图谱代码等信息一并录入, 生成供试雪茄种质的二维条码。

2 结果与分析

2.1 SSR引物的筛选

本研究利用6份烟草品种(Little Dutch、H211、Beinhart 1000-1、Hicks (Broad Leaf)、Havana 10、H382)对2000对候选引物进行筛选, 获得了43对多态性好、条带清晰的引物(表1)。

Table 1
表1
表1SSR 引物信息
Table 1Information of SSR markers

引物 Primer	上游引物 Forward primer (5'-3')	下游引物 Reverse primer (5'-3')	基因组 Group	位置 Position
PT52940	TCTTAGCCGTCCATCTCTGC	CACAGGACAATTTGTTGCAGTT	7	134.635
PT52118	ATCCCAAGCAAGTTGGTGTC	AACGAGCATTATGGGATTGC	18	70.312
PT51894	AACAACAGCGGATCACATTG	TGTATGCAACCCACAGAACAA	8	78.215
PT30163	CAACACACACGCTCATCAGA	TCCTCATCCATCGGTAAAGG	5	12.216
PT50433	TTTGTAAATTGTGTGTATAGAGGAGAG	GCACTGGATCTCCTGCATTT	16	160.451
PT51005	CCCTTCGTTCTACGAACCCT	GGGTGCCACCTCACTTTCT	10	55.231
PT51145	ATGCAAATGCAAGATGGTCA	CCGTTCAACTTGGCTTTACC	3	103.357
PT51234	AATTGGACCCAACCACTCTG	TAGCAGGGAAGCCGAAAGTA	19	66.109
PT51289	GAGTTGTGGCCAAGTAGCCT	GGCTTTAGCCAAACGCTCTA	21	49.700
PT51420	CGCCATTACGAACACTAGCA	AAGTTTAGTCCGGTCCGGTT	16	73.595
PT52821	GGGCAATGGAGCAATAGAAA	TGCTTAAAGATTTCCACTCGTT	20	95.174
PT53857	TTCATAAATCAACCTTAAGTGGAGC	TTTGGAGCTTGAAAGCAAAGA	22	67.708
PT54027	GGATCTGCTCATTTCCTATGG	CAAGGCAGCTAAGCCTAACAA	11	50.950
PT54444	GTGCCCATACCAGATTGTGA	ATTCTTCCAACATTCGCACC	6	54.463
PT55056	GCTCTTTAGTCTTTAGGGAGTTGG	TGAATTGAGCTCCTTCCAGG	14	0.276
PT60037	TGGTCTTTGTTCACCACCAA	GCTGGTTCCATCAATTCCAT	4	88.053
PT60146	AAACCTGGAAAGCAACATCA	GACCACGTTGTGTAGTTGTATTCTT	15	45.400
PT60379	AATTTGCACTCTGCGGACTC	CCCTGGGAGTTTCCATTTCT	14	28.140
PT60726	GCAGAGGCAACAATGCATAC	AGCTCGTTGCTGTTTCATCA	20	2.521
PT60990	TCAACCCATAAAGCTGCTCC	AAAGAGACAAAGCAGGCACAA	23	4.978
PT61187	GCTCAGTCGTGAAGAAACAGAA	AGGGAATCCTTGGTTGGTTT	5	130.242
PT61220	TTGGCATGGTTGTGTAAGGA	GCCACAAGTTATAATTTAAAGCTACCA	6	102.936
PT61226	CCTTGGACTCACAATACCATCA	CCTGTTAGCCAAAGGGTAATTC	13	46.719
PT30272	GAACCTAACCTCGCTCCACA	AAATGGTAGCTGCGAGGAGA	15	106.325
PT60427	TGATTCCCTAACCTGCGAAC	CCATTCTTGAGCTTCGTTCTTT	21	5.867
PT51378	ATGTCGGGAAGATTCCACAG	ACAACCAGCTAAATGTGCCC	17	111.136
PT1245	AGTTTAATGTGGGAGCGA	TCGGTCGGTATTAGTGGT	11	39.717
PT50736	CCAAGTAGCGACGAGGAGAA	TTATCCACGCGTCACCATTA	24	53.932
PT51059	CAAACCAGCGCATCTGTCTA	CACCAGAACCCGCTAGTCTC	18	15.309
PT51144	ACCGACAACACACGATTGTA	GCTTGTCGCTTAGCTTAACCAT	10	24.533
PT51878	TGGGAAGAGTGCTACCTTAGTACA	GCGATGTACTCCTAAAGATGTCAA	24	60.520
PT51976	CAGAGGGTAGTGAAGTATTCAATTT	GTTATCAATGCCCTGCCCT	22	94.306
PT52088	TGTGATTCGAACTTGTGATCCT	CATCAGTGGAAAGCGACTCA	7	87.079
PT52156	TGAGAATTCATATAACCGACCCA	TGCCTTCTTTCATACTGCCA	23	60.339
PT52176	TTGCATAAGAGTACCCATCTCG	TGTGTGTGTGAAGCTTGAAGAA	9	120.275
PT52689	GCACCAGGTGAAGTAAAGACAA	TGAAATGCATGTAAAGGAAAGG	13	66.991
PT53216	TCCAAACTCTTATCCCTGATCC	CACCTACCCTGTCGCTGTTT	2	0
PT54394	CATCTCAACTCGCCTGACAA	GCGGCAGGGTTCTATGATTA	8	96.735
PT54889	GGGTCTAAATAATCGCAATTGAA	CGCAGGTGGACTTAAAGAATC	19	106.147
PT60080	CTACGCAGAATCCAATTCCA	CATTAGGCCATAGCCATCCA	3	179.150
PT60917	CAGGAGACTCCATAAATTCACTCTT	AGCATGGTTGCTATTGGCTG	9	38.417
PT61025	GAACCAATCCAAGAGCCAGA	TCAGCCAAATATGTAGCAGGTG	1	87.176
PT61343	ACCCATTTAATTGCCACGTC	TATTCCGGCAGGAGTTTGAC	1	22.254

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2.2 遗传多样性分析

43对SSR引物在220份供试材料中共检测出243个基因位点(表2), 每对引物可检测的等位基因数(N_a)为2~13, 平均为5.65个, 其中PT52689的最多, PT52176的最少。有效等位基因数(N_e)在1.3081~11.7876之间, 平均为3.9077, 有效等位基因所占比例为69.15%。引物的多态性信息含量(PIC)在0.2078~0.9087之间, 平均约为0.6360。其中有35对引物的PIC值大于0.5, 为高度多态性信息引物, 说明所选引物的多态性普遍较高。观测杂合度(H_o)的变化范围为0.0828~0.7639, 平均值为0.3191; 群体平均Shannon’s遗传多样性指数(I)为1.3756, 其中PT52689的I最高, 为2.5112, 而PT52176的最低, 为0.3983; 平均Nei’s多样性指数(H)为0.6793。期望杂合度(H_e)的平均值为 0.6809, 最大是0.9172 (PT52689), 最小为0.2361 (PT52176)。H_e的平均值大于0.5, 说明供试雪茄烟种质的遗传多样性较高^[34]。

Table 2
表2
表243对SSR在220份材料中的遗传多样性参数
Table 2Genetic diversity of 43 SSR makers in 220 cigar germplasm resources

引物 Primer	等位基因数 Number of alleles (N_a)	有效等位基因数 Number of effective alleles (N_e)	Shannon’s 多样性指数 Shannon’s diversity index (I)	观测杂合度Observed heterozygosity (H_o)	期望杂合度Expected heterozygosity (H_e)	Nei’s 多样性指数 Nei’s diversity index (H)	多态性信息含量 Polymorphism information content (PIC)
PT52940	6	3.6152	1.3638	0.2750	0.7250	0.7234	0.6729
PT52118	4	2.9357	1.2000	0.3391	0.6609	0.6594	0.6019
PT51894	4	3.7568	1.3501	0.2645	0.7355	0.7338	0.6838
PT30163	4	2.2131	1.0189	0.4506	0.5494	0.5481	0.5017
PT50433	5	2.6611	1.0961	0.3743	0.6257	0.6242	0.5495
PT51005	7	5.0382	1.7676	0.1966	0.8034	0.8015	0.7760
PT51145	4	3.4657	1.3172	0.2869	0.7131	0.7115	0.6634
PT51234	4	1.5812	0.7298	0.6316	0.3684	0.3676	0.3415
PT51289	4	3.7381	1.3521	0.2658	0.7342	0.7325	0.6838
PT51420	4	2.9343	1.1847	0.3393	0.6607	0.6592	0.5943
PT52821	6	3.4023	1.3790	0.2923	0.7077	0.7061	0.6543
PT53857	6	1.8837	0.8822	0.5298	0.4702	0.4691	0.4117
PT54027	8	4.2139	1.6993	0.2355	0.7645	0.7627	0.7364
PT54444	3	2.4175	0.9805	0.4123	0.5877	0.5864	0.5170
PT55056	4	2.5613	1.1245	0.3890	0.6110	0.6096	0.5573
PT60037	5	2.4156	1.0418	0.4126	0.5874	0.5860	0.5153
PT60146	6	3.7451	1.4714	0.2653	0.7347	0.7330	0.6907
PT60379	6	4.4270	1.5781	0.2240	0.7760	0.7741	0.7375
PT60726	4	2.7073	1.1729	0.3679	0.6321	0.6306	0.5829
PT60990	5	4.3975	1.5329	0.2255	0.7745	0.7726	0.7352
PT61187	7	6.0233	1.8663	0.1641	0.8359	0.8340	0.8126
PT61220	6	4.8084	1.6639	0.2061	0.7939	0.7920	0.7621
PT61226	9	5.9444	1.9354	0.1663	0.8337	0.8318	0.8108
PT30272	7	5.6551	1.8289	0.1749	0.8251	0.8232	0.7999
PT60427	6	3.7898	1.5358	0.2622	0.7378	0.7361	0.7020
PT51378	7	5.5863	1.8098	0.1771	0.8229	0.8210	0.7969
PT1245	4	2.6871	1.0545	0.3707	0.6293	0.6279	0.5518
PT50736	3	2.1267	0.8116	0.4690	0.5310	0.5298	0.4192
PT51059	6	1.4182	0.6834	0.7045	0.2955	0.2949	0.2854
PT51144	8	5.2021	1.7598	0.1904	0.8096	0.8078	0.7804
PT51878	4	3.8172	1.3605	0.2603	0.7397	0.7380	0.6891
PT51976	9	6.7316	1.9878	0.1466	0.8534	0.8514	0.8335
PT52088	3	1.7058	0.7370	0.5853	0.4147	0.4138	0.3724
PT52156	4	2.0324	0.7439	0.4909	0.5091	0.5080	0.3878
PT52176	2	1.3081	0.3983	0.7639	0.2361	0.2355	0.2078
PT52689	13	11.7876	2.5112	0.0828	0.9172	0.9152	0.9087
PT53216	5	4.7386	1.5826	0.2092	0.7908	0.7890	0.7553
PT54394	4	2.1517	0.9554	0.4635	0.5365	0.5352	0.4776
PT54889	7	5.8341	1.8356	0.1695	0.8305	0.8286	0.8054
PT60080	10	7.0791	2.0931	0.1393	0.8607	0.8587	0.8433
PT60917	9	6.4282	1.9900	0.1536	0.8464	0.8444	0.8264
PT61025	5	2.7206	1.1786	0.3661	0.6339	0.6324	0.5793
PT61343	6	4.3419	1.5823	0.2285	0.7715	0.7697	0.7340
平均值Mean	5.6512	3.9077	1.3756	0.3191	0.6809	0.6793	0.6360

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2.3 种质间遗传距离

通过SSR标记分析, 220份雪茄烟种质材料间的遗传距离(genetic distance, GD)在0.0233~0.9286之间(表略), 平均为0.6816。其中良种(No.211)、辅善(No.212)和满耳朵(No.213) 3个材料, 山东大叶(No.75)和牡丹江05-1 (No.76) 2个材料, Florida 513 (No.125)和CA0701 (No.141) 2个材料间的遗传距离均为0, 即品种间差异位点数为0, 判定为疑同品种。古巴2号-2 (No.11)和古巴2号-3 (No.12)、Connecticut Broad Leaf (No.40)和CA0708 (No.147)、Havana 211 (No.17)和CA0709 (No.148)、2040 (No.67)和CS0707 (No.160)、Florida 301 (No.36)和florida30 (No.166)、松阳(No.206)和连选(No.207)材料间的遗传距离最小, 为0.0233, 品种间差异位点数为1, 判定为相近品种。这些材料有的是从同一国家的相近地区引进的, 如古巴2号-2 (No.11)和古巴2号-3 (No.12)、松阳(No.206)和连选(No.207); 有的原本是同一个品种在不同地区长期种植过程中被命名了不同的名称, 如Florida 513 (No.125)和CA0701 (No.141)、Connecticut Broad Leaf (No.40)和CA0708 (No.147)、Havana 211 (No.17)和CA0709 (No.148)、2040 (No.67)和CS0707 (No.160)、Florida 301 (No.36)和florida30 (No.166)。需结合田间表型调查结果对疑同品种和相近品种进行进一步鉴定, 从而确定是否为同一种质。Mont Calme Brun (No.63)和什新-2 (No.72)、万毛1号(No.82)和松阳(No.206)、万毛1号(No.82)和连选(No.207)的遗传距离最大, 达到0.9286, 说明两者的亲缘关系较远。材料间遗传距离的变化区间在0.8821左右, 平均遗传距离为0.6816, 表明供试材料间变异范围较大, 亲缘关系较远, 遗传差异性较显著。

2.4 参试种质的聚类分析

基于Nei’s 遗传距离, 利用UPGMA方法对220份雪茄烟材料进行聚类分析, 其结果(图1)表明, 利用SSR标记可以将220份雪茄烟资源相互区分开, 在GD=0.74时可将材料分为3个类群(I为第1类群, II、III、IV为第2类群, V、VI为第3类群); 而在遗传距离GD=0.70处, 其中第2类群又可细分为II、III、IV 三个小类, 第3类群又被划分为V、VI两个小类, 按此可将220份雪茄烟种质资源分为6个类群。类群I由万毛1号(No.82)、龙井香叶子(No.84)和牡晒89-23-1 (No.182) 3个国内晒烟和国外引进的贝拉烟(No.181)一共4个地方性晒烟品种组成。聚类分析表明, 这4个品种在遗传距离较大时(GD=0.76), 就能明显与其他品种区分开来, 可能是由于这些品种为地方性品种, 种植面积小, 从而保留了较丰富的遗传变异; 类群II由E18 (No.37)和Trapesond 288 (No.58) 2个从不同国家引进的品种组成; 类群III由3个国外引进品种Zrenjanin (No.32)、Copus Pobrecene No.1241 (No. 39)、Begej (No.43)和1个国内选育品种世纪一号(No.52)组成; 类群IV共有40个品种, 其中土耳其雪茄(No.3)、Manila (No.4)、Comstock Spanish (No.41)、Havana IIc (No.14)、Connecticut Broad Leaf (No.40)等37个国外引进品种, 和江西晒烟(No.205)、城口晒烟(No.208)、青州一号(No.134) 3个国内地方品种组成。美国康州阔叶(No.136)、Florida 513 (No.51)、古巴2号(No.9)、德雪1号(No.129)、德雪2号(No.130)、德雪3号(No.131)、Cubra-Brazil (No.135)、Habana92 (No.132)、Beinhart 1000-1 (No.42)、beinhart 1000-1 (No.165)、什邡枇杷柳(No.91)、吉县大烟叶子(No.100)等48个品种聚于类群V。印尼雪茄包皮(No.54)、Zimmer Spanish (No.31)、Segedinska Ruca (No.1)、哈瓦娜(No.13)、小牛舌(No.47)等35个雪茄烟品种, 和牡晒81-21-2 (No.179)、利川乌烟(No.77)、桐乡晒烟(No.79)、沂水大弯筋(No.70)等87个晾晒烟品种, 共122个品种聚于类群VI。从聚类图中可以发现, 我国的品种和从各国引进的雪茄烟品种并没有很明显的分开, 每个类群中的品种不是单一的来自同产地, 各国的品种分散在每个类群中, 这说明聚类结果与雪茄烟种质的地域来源并没有显著的相关性。

图1

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图1220份雪茄烟种质资源的SSR分析聚类图

类群I: 红色; 类群II: 蓝色; 类群III: 橙色; 类群IV: 湖蓝色; 类群V: 紫色; 类群VI: 绿色。
Fig. 1Cluster dendrogram of 220 cigar germplasm resources based on SSR markers

Group I: red; group II: blue; group III: orange; group IV: lake blue; group V: red; group VI: green.

2.5 主成分分析

根据SSR分子标记的结果用NTSYS软件根据位置近则亲缘关系近、位置远则亲缘关系远的原则^[46], 以第1主成分和第2主成分为二维坐标图的横坐标和纵坐标, 构建了220份雪茄烟种质资源的二维主坐标分析图。由图2可知, 220份雪茄烟种质材料划分为2个类群, 其中类群I包含36份种质, 类群II共有184份雪茄烟种质, 每个类群都含有不同地理来源的雪茄烟种质资源, 从PCA图中可以清晰的看出, PCA主成分分析与UPGMA聚类分析得出相同的结果, 即雪茄烟种质资源并没有按地理来源进化划分, 地理来源相同的种质资源交错分布于2个类群中。

图2

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图2220份雪茄烟种质资源的主成分分析(PCA)二维散点图

Fig. 2Two dimensional scatter plot of principal components analysis (PCA) from 220 cigar germplasm resources

2.6 遗传结构分析

利用Structure软件对220份雪茄烟种质的群体遗传结构分析中发现, 在K=1~15中, ln P(D)值随假定亚群数K值的增大而持续增大(图3-A), 则参照Evanno等^[43]的方法, 由ΔK值来确定220份雪茄烟资源的最优群体数K值, 当K=2时似然值最大(图3-B), 即所分析的雪茄烟种质从遗传结构上可划分为2个类群。参考苏一钧等^[47]的研究认为在structure群体遗传结构分析中, 当某一材料的Q值≥0.6表示该材料的遗传背景相对比较单一, 而当Q值<0.6, 则说明该材料的遗传背景较为复杂。分析我国雪茄烟种质资源的Q值发现, 有38份资源(No.24、No.28、No.30、No.31、No.32、No.36、No.38、No.39、No.43、No.50、No.56、No.75、No.76、No.81、No.83、No.87、No.98、No.104、No.105、No.114、No.118、No.125、No.131、No.137、No.141、No.155、No.158、No.164、No.165、No.166、No.172、No.179、No.183、No.191、No.192、No.196、No.218、No.220)的Q值<0.6, 说明这38份雪茄烟资源的遗传组成较为复杂, 没有明确归属于哪个类群, 形成一个混合群体, 其余的182份雪茄烟种质资源的Q值≥0.6, 此部分资源的遗传结构较为单一, 可以划在某一个类群中。220份雪茄烟种质资源的群体遗传结构如图4, 其中第I类群包括92份材料, 第II类群含有90份雪茄烟种质资源。所有的供试材料没有按照地理来源进行分类, 分析的结果与UPGMA聚类分析和PCA主成分分析的结果基本一致。

图3

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图3K值与ln P(D)、ΔK值折线图

A: ln P(D)值随K值变化折线图; B: ΔK值随K值变化折线图。
Fig. 3Line chart of K with ln P(D) and ΔK

A: line chart of ln P(D) with change of K-values; B: line chart of ΔK with change of K-values.

2.7 DNA指纹图谱的构建

本研究从43个SSR标记中选取14对SSR标记(PT52689、PT60080、PT51976、PT60917、PT61187、PT61226、PT54889、PT60726、PT51878、PT51145、PT52118、PT52821、PT60037、PT52940), 就可将供试雪茄烟烟种质完全区分开来。对于疑同品种和相近品种, 结合田间表型鉴定结果确定No.211、No.212和No.213三个材料, No.75和No.76两个材料, No.125和No.141两个材料为异名同种, 因此作为同一品种(No.211、No.212和No.213三个材料保留No.211; No.75和No.76两个材料保留No.75; No.125和No.141两个材料保留No.125)进行指纹图谱的构建。保留的216份雪茄烟种质资源指纹图谱代码构建方法依据之前设计的方案, 将14个引物依此用字母A~N表示, 每个引物生成的有差别的带型用该引物的字母和代表等位基因的数字编号表示, 为每份材料建立惟一能够区别于其他供试材料的指纹图谱代码(表3)。利用在线软件草料二维码生成器(http://cli.im/)对所有供试材料进行编码, 为每份材料构建了一份指纹图谱二维条码(图5), 其中包含了品种名称、全国统一编号、类型、植物学分类、指纹图谱代码等信息。如图1中A包含以下信息: 名称为 Segedinska Ruca, 全国统一编号为00001238, 类型为雪茄烟, 植物学分类为双子叶植物纲(Dicotyledoneae)管花目(Tubiflorae)茄科(Solanaceae)烟属(Nicotiana)普通烟草(Nicotiana tabacum L.), 指纹代码为A7B7C4D1E3F7G2H2I2J3K3L3M3N2。

图4

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图4基于SSR标记的220份雪茄烟种质资源群体遗传结构

Fig. 4Population structure of 220 cigar germplasm resources based on SSR markers

Table 3
表3
表3216份雪茄烟种质资源的SSR指纹图谱代码
Table 3SSR fingerprinting of 216 cigar germplasm resources

编号 No.	SSR指纹图谱 SSR fingerprinting	编号 No.	SSR指纹图谱 SSR fingerprinting
1	A7B7C4D1E3F7G2H2I2J3K3L3M3N2	110	A9B9C3D6E3F4G3H2I3J4K3L4M3N4
2	A7B7C4D1E3F7G2H3I2J3K3L3M3N2	111	A8B6C4D6E4F0G3H0I3J3K3L4M3N2
3	A10B2C7D4E4F8G5H2I1J1K1L3M1N3	112	A9B5C7D5E0F0G3H0I3J3K3L2M3N2
4	A10B2C6D4E4F8G5H4I1J1K1L3M1N3	113	A9B5C7D4E3F4G3H2I3J3K3L2M3N2
5	A2B4C5D4E2F8G5H4I4J1K2L2M1N3	114	A12B9C4D9E5F3G4H3I2J3K3L3M1N2
6	A2B4C5D4E2F8G5H4I4J1K2L3M1N3	115	A8B8C5D8E1F0G3H0I2J3K3L3M3N2
7	A8B7C5D4E4F2G2H4I3J4K2L4M3N1	116	A9B7C4D8E1F3G3H1I2J3K2L4M3N4
8	A8B2C5D4E7F2G2H1I3J4K2L3M2N1	117	A2B4C7D3E3F7G5H4I1J1K1L4M1N3
9	A1B7C3D4E7F5G3H2I4J4K2L4M2N1	118	A11B2C5D6E6F4G4H0I3J3K2L4M1N2
10	A1B2C3D3E3F5G3H2I4J4K2L4M1N1	119	A13B6C5D7E3F4G4H1I2J3K2L2M3N2
11	A1B2C3D3E3F5G3H2I4J4K2L4M2N1	120	A5B8C5D6E3F4G2H4I2J2K1L4M1N2
12	A1B2C3D3E3F5G3H2I4J1K2L4M2N1	121	A1B4C4D2E4F6G3H2I4J4K3L4M1N2
13	A7B5C5D4E4F5G1H2I2J4K3L2M3N1	122	A10B4C7D3E4F7G5H4I1J4K2L4M1N2
14	A3B4C6D3E3F9G5H4I4J1K3L2M1N3	123	A10B4C4D5E5F2G5H2I4J3K3L2M2N2
15	A3B4C6D3E3F9G1H4I2J1K3L2M1N3	124	A3B4C5D5E7F2G5H2I4J3K2L2M2N4
16	A12B4C3D3E4F5G5H4I1J1K1L4M1N3	125	A8B9C4D3E7F6G3H2I4J2K3L4M3N1
17	A4B4C8D3E3F9G5H4I1J1K1L4M1N4	126	A12B5C4D3E4F4G5H4I1J2K2L2M1N3
18	A5B7C7D3E4F9G5H3I2J1K3L4M1N1	127	A13B5C2D3E4F4G5H4I1J2K2L2M1N3
19	A12B4C4D3E4F4G4H4I1J1K1L2M1N3	128	A12B4C6D3E4F7G5H4I1J2K2L4M1N4
20	A2B4C8D3E1F9G4H4I4J1K3L2M1N3	129	A1B9C2D3E7F6G4H2I4J4K3L4M2N2
21	A2B4C6D3E1F9G4H4I4J1K3L2M1N3	130	A1B9C2D3E4F6G4H2I4J3K3L2M2N3
22	A7B5C5D4E4F5G1H3I2J4K3L2M3N1	131	A4B9C1D5E7F6G3H2I1J4K3L4M2N2
23	A2B3C2D3E2F4G3H3I2J1K3L2M1N2	132	A11B4C4D5E4F2G6H2I4J3K3L2M2N2
24	A2B2C6D5E4F5G1H3I4J3K1L5M1N3	133	A4B4C6D5E7F6G6H2I4J4K2L4M2N2
25	A2B2C7D6E7F4G5H3I4J3K1L2M3N4	134	A3B5C6D4E1F7G5H4I4J3K3L2M2N3
26	A8B2C7D5E7F5G5H1I4J2K1L2M1N2	135	A4B4C5D5E4F5G5H2I4J2K2L4M0N2
27	A8B9C2D6E3F4G2H2I2J3K3L3M3N1	136	A3B4C3D3E3F2G4H2I4J3K3L4M2N3
28	A3B7C5D2E7F7G3H2I3J3K3L3M3N2	137	A8B4C4D0E7F4G2H2I4J3K2L2M1N4
29	A4B2C6D2E3F8G4H4I1J1K3L2M1N3	138	A2B0C4D1E6F6G3H2I4J3K3L2M2N3
30	A7B2C6D6E6F5G2H2I3J1K3L6M1N4	139	A3B4C6D5E7F2G5H2I4J2K2L2M3N4
31	A1B7C2D2E5F6G4H2I3J4K3L2M3N2	140	A9B3C4D6E4F2G2H2I3J4K3L4M3N2
32	A13B6C8D5E3F3G2H4I3J3K4L6M1N4	142	A4B9C4D3E4F4G4H2I4J4K2L4M1N2
33	A7B2C6D6E6F5G2H2I4J1K3L2M1N3	143	A13B4C4D3E4F4G4H4I1J1K1L2M1N3
34	A12B3C4D3E7F3G2H4I3J3K1L2M1N2	144	A4B9C2D2E7F6G3H2I4J4K3L4M3N2
35	A2B2C7D3E1F8G5H4I4J2K3L3M1N4	145	A13B4C6D3E3F7G4H4I1J1K2L2M1N3
36	A8B2C5D5E3F4G2H2I3J2K3L2M3N2	146	A2B9C2D3E4F6G3H4I1J1K1L2M3N2
37	A7B2C4D4E7F6G2H4I3J1K3L2M3N4	147	A11B2C3D3E2F2G6H4I1J4K1L2M3N3
38	A2B6C4D3E1F3G4H2I3J3K4L5M3N2	148	A4B4C8D3E3F9G5H4I1J1K1L4M1N3
39	A3B7C6D2E4F4G6H1I3J3K4L5M1N1	149	A4B4C6D5E6F2G4H2I4J2K2L2M3N4
40	A11B2C3D3E2F2G5H4I1J4K1L2M3N3	150	A4B8C3D5E3F2G5H2I4J1K1L2M3N3
41	A11B4C4D3E4F5G6H4I1J1K2L2M1N4	151	A5B3C2D9E1F4G3H2I3J3K4L5M1N1
42	A6B2C5D6E7F5G1H3I4J2K3L5M1N4	152	A13B5C5D3E3F4G5H2I1J1K1L4M1N3
43	A10B2C7D2E4F1G2H3I3J4K3L3M1N4	153	A4B4C4D5E2F4G1H2I4J0K2L4M1N1
44	A1B3C4D4E7F6G4H3I2J4K3L3M3N4	154	A5B3C3D9E2F3G2H2I4J3K3L4M1N3
45	A7B7C2D5E5F6G3H1I2J4K3L2M3N1	155	A3B3C9D8E2F2G3H1I4J3K4L4M1N1
46	A6B8C1D7E4F2G7H1I2J4K3L1M3N4	156	A10B3C4D8E2F3G2H2I4J3K2L4M1N3
47	A9B7C1D8E4F3G1H1I2J4K3L3M3N1	157	A13B5C5D4E2F4G5H2I1J1K1L3M1N3
48	A4B2C3D5E5F5G4H3I4J4K3L2M1N4	158	A3B2C2D2E6F3G7H3I2J3K3L4M3N4
49	A6B6C2D9E3F6G1H2I2J4K3L3M3N1	159	A5B7C4D3E4F9G5H4I2J1K3L4M1N1
50	A10B5C6D6E4F2G5H4I1J4K3L2M1N1	160	A9B8C6D3E3F3G2H1I2J3K3L4M3N2
51	A8B7C3D4E7F5G3H3I4J2K3L3M3N1	161	A2B8C2D8E1F3G3H1I2J3K3L4M3N2
52	A10B6C1D4E4F7G2H1I2J2K3L3M3N4	162	A2B4C3D3E6F3G3H3I3J1K3L4M3N2
53	A8B2C3D5E4F2G2H2I3J3K3L2M3N2	163	A4B3C1D2E1F3G4H1I2J3K2L4M1N4
54	A8B2C2D6E4F2G1H2I3J3K3L4M3N2	164	A4B3C2D3E1F3G4H1I2J3K2L4M1N3
55	A10B2C7D4E7F2G4H2I1J1K3L4M3N2	165	A6B2C5D6E6F2G1H3I3J1K3L2M1N1
56	A4B2C2D4E7F1G4H2I4J1K3L4M2N2	166	A8B2C5D5E3F4G2H2I3J2K3L4M3N2
57	A5B4C6D4E2F7G4H2I3J2K3L4M2N2	167	A6B7C2D5E3F3G3H1I3J3K3L6M1N2
58	A8B2C2D5E7F2G2H1I4J1K4L4M3N2	168	A9B8C4D6E2F3G7H2I2J4K2L2M3N3
59	A4B2C8D4E4F2G3H2I2J4K3L5M2N2	169	A12B10C5D5E2F3G7H2I2J4K2L4M3N3
60	A2B3C3D4E4F3G3H2I2J2K3L4M3N2	170	A8B9C2D6E2F3G7H2I2J4K2L4M5N3
61	A3B1C5D3E4F3G5H2I4J1K2L3M3N3	171	A6B10C1D5E2F3G2H2I2J3K2L2M3N2
62	A4B2C7D3E0F0G6H0I2J2K1L0M2N3	172	A7B8C6D2E6F3G3H4I1J3K2L3M2N3
63	A4B2C7D3E1F4G6H4I2J2K2L3M2N2	173	A5B3C7D5E7F4G4H4I3J3K2L4M1N4
64	A13B3C4D3E1F4G6H4I2J2K1L2M2N2	174	A10B7C1D5E2F3G4H2I3J4K2L2M3N3
65	A12B3C4D3E1F4G6H4I2J2K1L4M2N2	175	A5B9C6D6E2F7G4H2I3J2K2L4M3N3
66	A12B3C4D3E2F4G5H4I1J2K1L2M2N3	176	A9B7C3D6E4F2G2H2I3J4K2L1M3N4
67	A8B8C6D3E3F3G2H1I2J3K3L4M3N2	177	A8B9C1D2E7F3G3H2I4J2K2L4M3N3
68	A3B1C7D6E7F3G5H3I2J1K2L4M3N3	178	A10B9C3D5E3F3G2H2I3J3K3L4M3N3
69	A3B1C7D3E2F3G5H3I2J1K2L4M3N3	179	A13B3C6D6E2F2G5H2I3J3K3L4M3N3
70	A5B4C2D8E2F3G4H2I3J3K1L4M3N3	180	A5B9C3D4E2F8G2H2I3J3K4L4M3N2
71	A8B7C4D6E2F3G2H2I3J3K3L4M3N3	181	A2B1C6D6E5F2G1H3I4J2K4L6M1N1
72	A6B7C3D8E3F5G7H2I3J4K3L4M5N4	182	A8B1C6D6E5F2G4H3I4J2K4L6M1N1
73	A8B6C3D8E3F5G7H2I3J4K3L4M5N4	183	A5B7C4D7E5F3G4H1I2J2K4L4M3N3
74	A6B7C3D8E4F3G7H2I3J4K3L4M3N4	184	A5B8C3D9E6F4G4H2I2J3K2L4M3N2
75	A11B6C7D7E3F3G4H4I3J3K1L3M1N2	185	A13B6C4D9E2F2G5H2I2J3K2L2M3N3
77	A6B8C2D6E3F3G2H2I3J3K4L4M3N4	186	A9B7C4D7E1F4G2H2I2J3K2L2M3N4
78	A1B8C2D7E4F3G4H2I4J3K3L4M3N2	187	A9B7C4D7E1F4G2H2I2J4K1L2M3N4
79	A2B7C3D8E6F4G2H2I2J3K3L6M3N2	188	A2B7C1D8E1F2G4H2I2J4K1L2M3N2
80	A7B7C7D8E7F3G7H2I3J3K3L3M3N4	189	A6B9C2D4E2F2G7H2I2J3K1L4M5N4
81	A1B2C4D3E6F4G5H2I3J1K3L3M1N4	190	A5B7C2D8E2F2G7H2I2J3K1L4M5N2
82	A11B2C4D2E3F3G6H3I4J2K4L3M1N3	191	A7B7C4D8E5F6G5H2I2J3K1L4M3N3
83	A2B7C7D8E7F5G3H2I2J1K3L5M3N3	192	A6B1C4D8E6F4G6H2I2J1K1L2M3N4
84	A5B7C3D8E3F4G3H2I2J2K3L3M1N5	193	A5B7C6D8E6F2G7H2I2J3K1L2M3N3
85	A5B9C5D8E2F5G3H4I2J3K3L2M3N4	194	A6B7C2D9E2F5G7H2I1J4K3L4M3N3
86	A6B4C3D5E3F4G3H2I3J3K3L3M3N4	195	A6B7C2D9E2F5G7H2I2J4K2L2M3N4
87	A3B9C3D3E4F4G4H3I3J3K3L3M1N2	196	A1B7C6D8E5F5G3H3I2J1K2L4M3N2
88	A9B7C1D9E3F4G3H2I1J3K3L5M3N3	197	A4B7C6D8E5F2G7H2I2J3K2L2M3N3
89	A2B2C4D3E6F3G4H2I2J1K3L4M3N3	198	A5B7C2D9E2F2G3H2I2J3K3L2M3N2
90	A10B7C4D3E3F4G5H2I1J3K3L3M3N2	199	A5B7C2D9E1F2G3H2I2J3K2L2M3N2
91	A4B2C2D3E5F8G4H3I3J2K3L4M3N4	200	A5B7C2D9E1F2G2H2I2J3K2L2M3N2
92	A9B8C5D9E3F4G3H2I2J3K3L3M4N2	201	A5B9C2D8E2F1G2H2I2J3K2L1M3N3
93	A9B7C5D8E3F4G3H2I2J3K3L4M3N2	202	A5B7C2D9E2F5G7H3I2J4K2L2M3N3
94	A10B7C2D8E3F4G1H2I2J3K3L6M3N2	203	A5B7C2D9E2F2G2H2I2J3K3L2M3N2
95	A8B8C5D8E3F4G3H2I3J3K3L2M3N2	204	A5B7C2D9E2F2G2H2I1J3K2L2M3N2
96	A8B8C2D6E3F4G3H2I1J3K3L2M3N2	205	A13B2C4D3E4F5G6H3I1J1K1L2M1N3
97	A6B6C3D6E3F4G3H2I3J3K3L6M1N1	206	A2B7C2D8E6F4G2H2I1J3K3L6M3N2
98	A4B7C7D6E3F4G3H3I4J3K4L2M3N4	207	A2B7C2D8E6F4G2H2I1J3K2L6M3N2
99	A12B8C3D6E3F1G5H2I1J3K3L5M3N4	208	A1B2C6D3E1F8G5H4I3J2K2L3M1N3
100	A3B2C5D3E5F6G5H3I4J1K3L3M1N4	209	A1B1C6D6E6F2G6H3I3J2K1L3M3N4
101	A9B6C4D6E3F4G4H2I3J3K4L4M3N4	210	A2B1C6D6E6F2G6H3I3J2K1L3M3N4
102	A8B9C2D6E3F5G7H2I3J3K3L2M3N3	211	A6B2C5D3E4F4G7H2I3J3K2L3M3N2
103	A9B5C3D4E1F5G4H2I3J0K0L2M3N4	214	A3B2C2D7E4F4G7H2I0J3K2L3M3N2
104	A4B2C2D6E5F0G4H3I3J1K3L4M3N4	215	A1B7C7D5E7F3G4H3I4J2K2L4M3N2
105	A8B6C6D5E3F4G5H3I1J2K3L3M1N2	216	A5B6C4D9E5F4G2H2I4J4K2L2M3N3
106	A9B8C3D9E3F4G4H3I3J3K3L3M3N2	217	A3B1C8D6E6F5G5H3I0J0K4L4M3N3
107	A10B9C3D6E3F4G4H3I3J2K3L4M3N4	218	A1B1C2D4E4F3G3H1I4J3K2L4M1N2
108	A9B9C8D6E1F5G4H2I3J2K3L2M3N4	219	A1B1C8D6E5F5G5H3I4J2K4L4M3N3
109	A10B8C8D8E4F0G3H0I3J4K3L0M3N4	220	A1B1C2D4E4F3G3H1I4J3K2L4M1N7

The number in Table 4 corresponds to the same number in Table 1; A-N represent primer of SSR PT52689, PT60080, PT51976, PT60917, PT61187, PT61226, PT54889, PT60726, PT51878, PT51145, PT52118, PT52821, PT60037, PT52940, respectively.
表4中编号对应表1同编号品种; A~N分别代表SSR引物PT52689、PT60080、PT51976、PT60917、PT61187、PT61226、PT54889、PT60726、PT51878、PT51145、PT52118、PT52821、PT60037、PT52940。

新窗口打开|下载CSV

图5

新窗口打开|下载原图ZIP|生成PPT
图5部分雪茄烟品种指纹图谱二维条码

A~F分别为雪茄烟品种Segedinska Ruca、Scafati、土耳其雪茄、Manila、Connecticut Shade和Bad Geudertheimer Landsorte。
Fig. 5Fingerprinting of 2D barcode from part cigar varieties A-F represent the melon samples

Segedinska Ruca, Scafati, Turkey Cigar, Manila, Connecticut Shade, and Bad Geudertheimer Landsorte, respectively.

3 讨论

3.1 雪茄烟种质资源遗传多样性分析

进行雪茄烟种质资源遗传多样性的研究对我国雪茄烟种质的鉴定、新基因的挖掘、新品种选育等育种工作至关重要。目前有不少烟草方面遗传多样性研究的报道, 但多为烤烟和晾晒烟, 而雪茄烟只进行过形态学上的鉴定^[48,49], 受外界环境条件和栽培措施的影响较大, 极易产生误差导致结果不准确。作物品种鉴定的最可靠方法是品种的基因型鉴定, 利用分子标记技术从DNA水平上区分鉴别雪茄烟基因型间的遗传差异, 不受外界环境条件和栽培措施的影响, DNA分析方法是目前为止最有效的遗传分析方法^[50]。操作简单、多态性好、耗费低的SSR标记已逐渐取代存在诸多缺点的通用型DNA标记RAPD、AFLP等。SSR标记已被广泛应用于烟草种质资源与品种遗传多样性分析^{[4,20-22,51-52]}, 因其成本较低也被视为烟草品种鉴定的理想工具^[18-19,53]。

本研究利用SSR标记首次分析了雪茄烟种质资源遗传多样性, 因此, 没有与之相关的研究报道可供参考和比较。本研究利用43对SSR分子标记对收集到的220份雪茄烟种质资源的遗传多样性进行分析发现, 平均等位基因数为5.65, H为0.6793, PIC为0.6360, 说明供试种质具有较高的遗传多样性, 引物总体上有较好的鉴别能力。供试材料间的遗传距离(GD)在0.0233~0.9286之间, 平均为0.6816, 说明雪茄烟种质资源的遗传距离较大, 遗传多样性比较丰富。主要原因可能为: 虽然我国最近十年才开始进行雪茄烟的大规模引种和栽培, 但是晾晒烟的种植历史悠久, 我国使用优质地方晾晒烟品种当做茄芯来使用, 当地也利用地方晾晒烟品种和引进的各种雪茄烟品种进行杂交, 以获得品种改良, 这在无形中拓宽了我国雪茄烟种质资源的遗传多样性。在220份雪茄烟种质资源中, 良种(No.211)、辅善(No.212)和满耳朵(No.213)、山东大叶(No.75)和牡丹江05-1 (No.76)、Florida 513 (No.125)和CA0701 (No.141)等种质被判定为疑同品种; 古巴2号-2 (No.11)和古巴2号-3 (No.12)、Connecticut Broad Leaf (No.40)和CA0708 (No.147)、Havana 211 (No.17)和CA0709 (No.148)、2040 (No.67)和CS0707 (No.160)、Florida 301 (No.36)和florida30 (No.166)、松阳(No.206)和连选(No.207)等种质被判定为相近品种, 具有非常高的遗传相似性。结合田间表型性状鉴定结果发现, 辅善(No.212)和满耳朵(No.213)、山东大叶(No.75)和牡丹江05-1 (No.76)、Florida 513 (No.125)和CA0701 (No.141)等种质为同一品种, 即异名同种。由此可见, SSR标记适用于雪茄烟品种鉴定和遗传多样性的研究中。SSR标记在雪茄烟遗传多样性分析、种质资源品种鉴定方面的应用, 为雪茄烟新品种的管理、杂交育种、产权保护提供科学依据。

本研究通过UPGMA聚类分析、PCA主成分分析和Structure软件的遗传结构分析3种分析方法对供试材料进行划分, 三者在分类结果上存在差异: UPGMA聚类分析在GD=0.74处将供试材料划分为3个大类, 而群体遗传结构和PCA主成分分析则将供试材料划分为2个亚群; UPGMA聚类分析和PCA主成分分析在类群内品种分布的结果上高度一致, 而群体遗传结构分析结果与这两者在类群内品种的分布上存在较大差异。造成差异的主要原因可能是各自的分类依据不同: 聚类分析和PCA主成分分析分别依据的是遗传距离(GD)和遗传相似系数(GS), 反映的都是品种间亲缘关系的远近, 这2种分析方法在类群内具体品种的分布结果上高度一致; 而群体遗传结构分类的依据服从Hardy-Weinberg平衡的亚群数目, 是建立在数学模型的基础上, 并计算各供试材料间相应的Q值。因此, Structure软件的遗传结构分析的结果避免了人为因素对群体划分的影响, 具有较高的准确性^[34]。3种分析方法的结果均显示, 收集的这220份来自各地的雪茄烟资源交错分布于每个类群中, 遗传分类与地域因素没有显著相关性。主要因为我国是从引种鉴定、推广适宜各产区栽培条件的优良品种开始雪茄烟种植的, 并在此基础上发展新的雪茄烟品种。我国的雪茄烟资源大多是从国外引进, 有些雪茄烟资源有较好的香气, 如Havana系列品种; 有的是好的抗源, 如高抗黑胫病的Florida301, 高抗黑胫病、赤星病的Beinhart 1000-1等, 这些优质、抗病品种被广泛用作亲本与其他不同品种进行杂交选育出品质较好、抗病能力强的品种或是作为试验材料进行雪茄烟分子育种研究, 也进一步表明我国雪茄烟种质资源的遗传多样性丰富。本研究中的220份雪茄烟种质资源的聚类结果与地理来源无相关关系, 说明我国雪茄烟种质资源遗传背景比较复杂。

3.2 雪茄烟种质资源指纹图谱库的构建

DNA指纹图谱是在分子标记的基础上绘制而成的, 这种图谱多态性丰富, 具有高度的个体特异性和环境稳定性, 犹如人的指纹一样能够鉴别生物个体之间的差异, 因此被称为DNA指纹图谱(DNA fingerprinting)^[54]。DNA指纹图谱具有快速、准确的特点, 是鉴别品种、品系的有力工具, 也非常适应于种质资源的鉴定工作^[55]。SSR分子标记特异性强、谱带清晰、数据准确, 适合大量资源的指纹图谱构建工作^[35], 利用SSR标记进行烟草种质资源指纹图谱的构建已有报道^[35,37,56]。本研究利用筛选出的14对可以完全区分所有供试材料的SSR引物, 和根据供试材料分别在14对引物扩增出条带的分子量大小及条带类型构建了由字母和阿拉伯数字组成的216份雪茄烟种质资源DNA指纹图谱代码; 结合供试材料的相关信息和指纹图谱代码进行编码又构建了216份雪茄烟种质资源的DNA指纹图谱二维条码。本研究通过这2种方式对雪茄烟种质资源进行指纹图谱库的构建, 使每个品种都有唯一的一套DNA指纹, 可在分子水平上解决同种异名和同名异种等品种混乱现象, 若2份材料的指纹数据一致则被认为是同一品种。雪茄烟种质资源SSR指纹图谱库的建立使品种鉴定更加高效准确, 为以后雪茄烟种质资源的鉴定、保护及育种研究奠定理论基础。

4 结论

本研究利用43对SSR引物对220份雪茄烟种质资源进行遗传多样性分析表明, 这些材料亲缘关系较远, 遗传差异性显著, 遗传多样性丰富, 这对我国雪茄烟种质资源的利用及品种选育方面具有重要意义; 利用从43对SSR引物中筛选出的14对核心引物构建了216份雪茄烟种质资源的指纹图谱库, 为今后我国雪茄烟品种鉴定体系的研究提供技术支撑。

附表请见网络版: 1) 本刊网站http://zwxb.chinacrops.org/; 2) 中国知网http://www.cnki.net/; 3) 万方数据http://c.wanfangdata.com.cn/Periodical-zuowxb.aspx。

Table S1
附表1
附表1 供试的烟草种质资源材料
Table S1 Tobacco germplasm resources

编号No.	种质名称 Germplasm name	类型 Type	种质提供单位 Origin	来源 Source
1	Segedinska Ruca	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	南斯拉夫 Yugoslavia
2	Scafati	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	意大利 Italy
3	土耳其雪茄 Turkish cigar	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	土耳其 Turkey
4	Manila	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	菲律宾 the Philippines
5	Connecticut Shade	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	美国 USA
6	Bad Geudertheimer Landsorte	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	德国 Germany
7	H423	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	美国 USA
8	2042	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)
9	古巴2号 Cuba 2	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	古巴 Cuba
10	古巴2号-1 Cuba 2-1	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	古巴 Cuba
11	古巴2号-2 Cuba 2-2	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	古巴 Cuba
12	古巴2号-3 Cuba 2-3	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	古巴 Cuba
13	哈瓦娜 Hawana	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	古巴 Cuba
14	Havana IIc	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	波兰 Poland
15	Havana IIc	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	德国 Germany
16	Havana10	雪茄烟 Cigar tobacco	国家烟草种质资源中期库National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	波兰 Poland
17	Havana 211	雪茄烟 Cigar tobacco	国家烟草种质资源中期库National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	美国 USA
18	H211	雪茄烟 Cigar tobacco	国家烟草种质资源中期库National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	古巴 Cuba
19	Havana 38	雪茄烟 Cigar tobacco	国家烟草种质资源中期库National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	德国 Germany
20	Havana 510	雪茄烟 Cigar tobacco	国家烟草种质资源中期库National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	波兰 Poland
21	Havana Connecticut	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	美国 USA
22	夏湾那 Xiawanna	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	古巴 Cuba
23	Criollo Salteno 11	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	巴西 Brazil
24	多米尼加短芯 Dominican Short Core Leaf	雪茄烟 Cigar tobacco	德阳市烟草公司 Deyang Tobacco Company	多米尼加 Dominican
25	多米尼加长芯Dominican Long Core Leaf	雪茄烟 Cigar tobacco	德阳市烟草公司 Deyang Tobacco Company	多米尼加 Dominican
26	尼加拉瓜短芯 Nigaragua Short Core Leaf	雪茄烟 Cigar tobacco	德阳市烟草公司 Deyang Tobacco Company	尼加拉瓜 Nicaragua
27	H382	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	印度尼西亚 Indonesia
28	J 20	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)
29	Little Dutch	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	美国 USA
30	Oosikappal	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)
31	Zimmer Spanish	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	西班牙 Spain
32	Zrenjanin	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	南斯拉夫 Yugoslavia
33	铁杆青 Tieganqing	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国贵州 Guizhou, China
34	Sumatra Mitlingskaja	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	印度尼西亚 Indonesia
35	Friedrichstaier	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	德国 Germany
36	Florida 301	雪茄烟 Cigar tobacco	国家烟草种质资源中期库National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	美国 USA
37	E 18	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	德国 Germany
38	Dirojuio Fagues No.361	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	波兰 Poland
39	Copus Pobrecene No.1241	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	波兰 Poland
40	Connecticut Broad Leaf	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	美国 USA
41	Comstock Spanish	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	西班牙 Spain
42	Beinhart 1000-1	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	美国 USA
43	Begej	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	南斯拉夫 Yugoslavia
44	422 (Kedoe Hybrid 43)	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)
45	112-117	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	美国 USA
46	什邡70-30号 Shifang70-30	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国四川 Sichuan, China
47	小牛舌 Xiaoniushe	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国江西 Jiangxi, China
48	Conn Shade	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	美国 USA
49	Bel-W3	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	美国 USA
50	诺凡斯瑞加 Nuofansiruijia	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)
51	Florida 513	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	美国 USA
52	世纪1号 Shiji 1	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国浙江 Zhejiang, China
53	2052	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)
54	印尼雪茄包皮 Indonesia cigar wrapper	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	印度尼西亚 Indonesia
55	印尼02号 Indonesia 02	雪茄烟 Cigar tobacco	国家烟草种质资源中期库National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	印度尼西亚 Indonesia
56	无名雪茄 Wuming cigar	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)
57	Trapezund 610	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	波兰 Poland
58	Trapesond 288	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	俄罗斯 Russia
59	Sumatra Deli	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	印度尼西亚 Indonesia
60	Pennbel 69	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	美国 USA
61	Pennsylvania Seed Leaf	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	美国 USA
62	Philippin	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	菲律宾 Philippines
63	Mont Calme Brun	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)
64	Lanka 23	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)
65	Hanica	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	德国 Germany
66	Geudetthelmex	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	德国 Germany
67	2040	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)
68	Burley 21	白肋烟 Burley	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	美国 USA
69	Burley 37	白肋烟 Burley	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	美国 USA
70	沂水大弯筋 Yishuidawanjin	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国山东 Shandong, China
71	塘蓬 Tangpeng	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国广东 Guangdong, China
72	什新-2 Shixin-2	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国四川 Sichuan, China
73	什新-1 Shixin-1	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国四川 Sichuan, China
74	宣汉晒烟76-2 Xuanhanshaiyan 76-2	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国四川 Sichuan, China
75	山东大叶 Shandongdaye	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国山东 Shandong, China
76	牡丹江05-1 Mudanjiang 05-1	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国黑龙江 Heilongjiang, China
77	利川乌烟 Lichuanwuyan	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国湖北 Hubei, China
78	罗三湾青毛烟 Luosanwanqingmaoyan	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国湖北 Hubei, China
79	桐乡晒烟 Tongxiangshaiyan	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国浙江 Zhejiang, China
80	NY-3	晒烟 Sun-cured tobacco	德阳市烟草公司 Deyang Tobacco Company	印度尼西亚 Indonesia
81	公会晒烟 Gonghuishaiyan	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国广西 Guangxi, China
82	万毛1号 Wanmao 1	晒烟 Sun-cured tobacco	德阳市烟草公司 Deyang Tobacco Company	中国重庆 Chongqing, China
83	万毛2号 Wanmao 2	晒烟 Sun-cured tobacco	德阳市烟草公司 Deyang Tobacco Company	中国重庆 Chongqing, China
84	龙井香叶子 Longjingxiangyezi	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国吉林 Jilin, China
85	凤凰香烟 Fenghuangxiangyan	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国湖南 Hunan, China
86	黄苗2220 Huangmiao 2220	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国河南 Henan, China
87	新宾小团叶 Xinbinxiaotuanye	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国辽宁 Liaoning, China
88	督叶尖杆种 Duyejianganzhong	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国浙江 Zhejiang, China
89	红花铁杆子 Honghuatieganzi	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国四川 Sichuan, China
90	尚志一朵花 Shangzhiyiduohua	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国黑龙江 Heilongjiang, China
91	什邡枇杷柳 Shifangpipaliu	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国四川 Sichuan, China
92	伟俄小柳叶 Weiexiaoliuye	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国贵州 Guizhou, China
93	铁赤烟 Tiechiyan	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国江西 Jiangxi, China
94	丹阳烟 Danyangyan	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国江苏 Jiangsu, China
95	人和烟 Renheyan	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国广东 Guangdong, China
96	云罗03 Yunluo 03	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国广东 Guangdong, China
97	稀格巴小黑烟 Xigebaxiaoheiyan	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国贵州 Guizhou, China
98	辽多叶 Liaoduoye	晒烟 Sun-cured tobacco	国家烟草种质资源中期库National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国辽宁 Liaoning, China
99	江油烟 Jiangyouyan	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国四川 Sichuan, China
100	吉县大烟叶子 Jixiandayanyezi	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国山西 Shanxi, China
101	巫山小兰烟 Wushanxiaolanyan	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国四川 Sichuan, China
102	湄潭晒烟 Meitanshaiyan	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国贵州 Guizhou, China
103	鹤峰黑烟 Hefengheiyan	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国湖北 Hubei, China
104	铁板青 Tiebanqing	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国湖北 Hubei, China
105	泌阳晒烟(4) Biyangshaiyan (4)	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国河南 Henan, China
106	六峰毛把烟 Liufengmaobayan	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国湖北 Hubei, China
107	阳和大乌烟 Yanghedawuyan	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国湖北 Hubei, China
108	建平瓢把烟 Jianpingpiaobayan	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国四川 Sichuan, China
109	巫山大南烟-1 Wushandananyan-1	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国四川 Sichuan, China
110	早阳人头烟 Zaoyangrentouyan	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国四川 Sichuan, China
111	丛树大青烟 Congshudaqingyan	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国重庆 Chongqing, China
112	大叶子青烟 Dayeziqingyan	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国重庆 Chongqing, China
113	巫溪火烟-2 Wuxihuoyan-2	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国重庆 Chongqing, China
114	利川毛烟 Lichuanmaoyan	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国湖北 Hubei, China
115	竹山大柳子 Zhushandaliuzi	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国湖北 Hubei, China
116	川柳子烟 Chuanliuziyan	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国四川 Sichuan, China
117	Wisconsin38	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	美国 USA
118	Q-5-0082	药烟 Herbal medicine tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国山西 Shanxi, China
119	81-8-6	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国广东 Guangdong, China
120	A-0014	药烟 Herbal medicine tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国山西 Shanxi, China
121	Havana-3	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	古巴 Cuba
122	Havana-1	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	古巴 Cuba
123	Havana 1	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	古巴 Cuba
124	S-2	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	古巴 Cuba
125	Florida 513	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	美国 USA
126	Geudetthelmex	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	德国 Germany
127	Havana 10	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	波兰 Poland
128	土耳其雪茄烟 Turkish cigar	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	土耳其 Turkey
129	德雪1号 Dexue 1	雪茄烟 Cigar tobacco	德阳市烟草公司 Deyang Tobacco Company	古巴 Cuba
130	德雪2号 Dexue 2	雪茄烟 Cigar tobacco	德阳市烟草公司 Deyang Tobacco Company	古巴 Cuba
131	德雪3号 Dexue 3	雪茄烟 Cigar tobacco	德阳市烟草公司 Deyang Tobacco Company	古巴 Cuba
132	Habana92	雪茄烟 Cigar tobacco	德阳市烟草公司 Deyang Tobacco Company	古巴 Cuba
133	C-15	雪茄烟 Cigar tobacco	德阳市烟草公司 Deyang Tobacco Company	古巴 Cuba
134	青州一号 Qingzhou 1	雪茄烟 Cigar tobacco	德阳市烟草公司 Deyang Tobacco Company	中国 China
135	Cubra-Brazil	雪茄烟 Cigar tobacco	德阳市烟草公司 Deyang Tobacco Company	巴西 Brazil
136	美国康州阔叶 America Connecticut Broadleaf	雪茄烟 Cigar tobacco	德阳市烟草公司 Deyang Tobacco Company	美国 USA
137	OLOR	雪茄烟 Cigar tobacco	德阳市烟草公司 Deyang Tobacco Company	多米尼加 Dominican
138	来凤H382 Laifeng H382	雪茄烟 Cigar tobacco	德阳市烟草公司 Deyang Tobacco Company	中国来凤 Laifeng, China
139	海南-1 Hainan-1	雪茄烟 Cigar tobacco	德阳市烟草公司 Deyang Tobacco Company	中国海南 Hainan, China
140	海南-2 Hainan-2	雪茄烟 Cigar tobacco	德阳市烟草公司 Deyang Tobacco Company	中国海南 Hainan, China
141	CA0701	雪茄烟 Cigar tobacco	德阳市烟草公司 Deyang Tobacco Company	中国浙江 Zhejiang, China
142	CA0702	雪茄烟 Cigar tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	美国 USA
143	CA0703	雪茄烟 Cigar tobacco	德阳市烟草公司 Deyang Tobacco Company	美国 USA
144	CA0704	雪茄烟 Cigar tobacco	德阳市烟草公司 Deyang Tobacco Company	美国 USA
145	CA0705	雪茄烟 Cigar tobacco	德阳市烟草公司 Deyang Tobacco Company	波兰 Poland
146	CA0706	雪茄烟 Cigar tobacco	德阳市烟草公司 Deyang Tobacco Company	德国 Germany
147	CA0708	雪茄烟 Cigar tobacco	德阳市烟草公司 Deyang Tobacco Company	美国 USA
148	CA0709	雪茄烟 Cigar tobacco	德阳市烟草公司 Deyang Tobacco Company	波兰 Poland
149	尼加拉瓜长芯 Nicaragua Long Core Leaf	雪茄烟 Cigar tobacco	德阳市烟草公司 Deyang Tobacco Company	尼加拉瓜 Nicaragua
150	哈珀纳斯 Haponasi	雪茄烟 Cigar tobacco	德阳市烟草公司 Deyang Tobacco Company	古巴 Cuba
151	MFPP	雪茄烟 Cigar tobacco	德阳市烟草公司 Deyang Tobacco Company	巴西 Brazil
152	MFZS	雪茄烟 Cigar tobacco	德阳市烟草公司 Deyang Tobacco Company	巴西 Brazil
153	CP2011	雪茄烟 Cigar tobacco	德阳市烟草公司 Deyang Tobacco Company	巴西 Brazil
154	MSCA	雪茄烟 Cigar tobacco	德阳市烟草公司 Deyang Tobacco Company	多米尼加 Dominica
155	MNC	雪茄烟 Cigar tobacco	德阳市烟草公司 Deyang Tobacco Company	多米尼加 Dominica
156	DM长 DM Chang	雪茄烟 Cigar tobacco	德阳市烟草公司 Deyang Tobacco Company	多米尼加 Dominica
157	CS0701	雪茄烟 Cigar tobacco	德阳市烟草公司 Deyang Tobacco Company	菲律宾 the Philippines
158	CS0704	雪茄烟 Cigar tobacco	德阳市烟草公司 Deyang Tobacco Company	中国广东 Guangdong, China
159	CS0705	雪茄烟 Cigar tobacco	德阳市烟草公司 Deyang Tobacco Company	中国广东 Guangdong, China
160	CS0707	雪茄烟 Cigar tobacco	德阳市烟草公司 Deyang Tobacco Company	中国四川 Sichuan, China
161	CS0708	雪茄烟 Cigar tobacco	德阳市烟草公司 Deyang Tobacco Company	中国山东 Shandong, China
162	CS0709	雪茄烟 Cigar tobacco	德阳市烟草公司 Deyang Tobacco Company	中国四川 Sichuan, China
163	CS0713	雪茄烟 Cigar tobacco	德阳市烟草公司 Deyang Tobacco Company	中国四川 Sichuan, China
164	3763	雪茄烟 Cigar tobacco	德阳市烟草公司 Deyang Tobacco Company	中国四川 Sichuan, China
165	beinhart1000-1	雪茄烟 Cigar tobacco	德阳市烟草公司Deyang Tobacco Company	美国 USA
166	florida30	雪茄烟 Cigar tobacco	德阳市烟草公司 Deyang Tobacco Company	美国 USA
167	稀格巴小黑烟 Xigebaxiaoheiyan	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国贵州 Guizhou, China
168	高产烟-3 Gaochanyan-3	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国广东 Guangdong, China
169	包盖烟-1 Baogaiyan-1	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国广东 Guangdong, China
170	四川青毛 Sichuanqingmao	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国四川 Sichuan, China
171	青毛籽 Qingmaozi	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国四川 Sichuan, China
172	四川黄毛 Sichuanhuangmao	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国四川 Sichuan, China
173	奉节大毛烟 Fengjiedamaoyan	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国重庆 Chongqing, China
174	大火柳子烟 Dahuoliuziyan	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国湖北 Hubei, China
175	黄金烟 Huangjinyan	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国湖南 Hunan, China
176	青梗8 Qinggeng 8	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国广东 Guangdong, China
177	泡杆烟 Paoganyan	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国贵州 Guizhou, China
178	云南荼科所晒烟 Yunnantukesuoshanyan	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国云南 Yunnan, China
179	牡晒81-21-2 Mushai 81-21-2	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国黑龙江 Heilongjiang, China
180	州852 Zhou 852	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国湖南 Hunan, China
181	贝拉烟 Beilayan	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	几内亚 Guinea
182	牡晒89-23-1 Mushai 89-23-1	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国黑龙江 Heilongjiang, China
183	晒9118 Shai 9118	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国湖南 Hunan, China
184	81-8-6	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国广东 Guangdong, China
185	清远牛利 Qingyuanniuli	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国广东 Guangdong, China
186	葵烟 Kuiyan	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国广东 Guangdong, China
187	湘潭细叶枇杷 Xiangtanxiyepipa	晒烟 Sun-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingd ao)	中国湖南 Hunan, China
188	什烟一号 Shiyan No.1	晒烟 Sun-cured tobacco	德阳市烟草公司 Deyang Tobacco Company	中国四川 Sichuan, China
189	葵柳 Kuiliu	晒烟 Sun-cured tobacco	德阳市烟草公司 Deyang Tobacco Company	中国四川 Sichuan, China
190	笆毛柳 Bamaoliu	晒烟 Sun-cured tobacco	德阳市烟草公司 Deyang Tobacco Company	中国四川 Sichuan, China
191	绵竹晒烟 Mianzhushaiyan	晒烟 Sun-cured tobacco	德阳市烟草公司 Deyang Tobacco Company	中国四川 Sichuan, China
192	GW3	晒烟Sun-cured tobacco	德阳市烟草公司 Deyang Tobacco Company	中国四川 Sichuan, China
193	MJ1	晒烟Sun-cured tobacco	德阳市烟草公司 Deyang Tobacco Company	中国四川 Sichuan, China
194	MJ2	晒烟 Sun-cured tobacco	德阳市烟草公司 Deyang Tobacco Company	中国四川 Sichuan, China
195	巴山一号 Bashan 1	晒烟 Sun-cured tobacco	德阳市烟草公司 Deyang Tobacco Company	中国四川 Sichuan, China
196	万毛3号 Wanmao 3	晒烟 Sun-cured tobacco	德阳市烟草公司 Deyang Tobacco Company	中国重庆 Chongqing, China
197	崇州晒烟1 Chongzhoushaiyan 1	晒烟 Sun-cured tobacco	德阳市烟草公司 Deyang Tobacco Company	中国四川 Sichuan, China
198	崇州晒烟2 Chongzhoushaiyan 2	晒烟 Sun-cured tobacco	德阳市烟草公司 Deyang Tobacco Company	中国四川 Sichuan, China
199	州烟 Zhouyan	晒烟 Sun-cured tobacco	德阳市烟草公司 Deyang Tobacco Company	中国四川 Sichuan, China
200	铁杆子 Tieganzi	晒烟 Sun-cured tobacco	德阳市烟草公司 Deyang Tobacco Company	中国四川 Sichuan, China
201	黄毛烟 Huangmaoyan	晒烟 Sun-cured tobacco	德阳市烟草公司 Deyang Tobacco Company	中国四川 Sichuan, China
202	白毛烟 Baimaoyan	晒烟 Sun-cured tobacco	德阳市烟草公司 Deyang Tobacco Company	中国四川 Sichuan, China
203	杂交晒烟 Zajiaoshaiyan	晒烟 Sun-cured tobacco	德阳市烟草公司 Deyang Tobacco Company	中国四川 Sichuan, China
204	白花 Bahua	晒烟 Sun-cured tobacco	德阳市烟草公司 Deyang Tobacco Company	中国河南 Henan, China
205	江西晒烟 Jiangxishaiyan	晒烟 Sun-cured tobacco	德阳市烟草公司 Deyang Tobacco Company	中国江西 Jiangxi, China
206	松阳晒烟 Songyangshaiyan	晒烟 Sun-cured tobacco	德阳市烟草公司 Deyang Tobacco Company	中国浙江 Zhejiang, China
207	连选晒烟 Lianxuanshaiyan	晒烟 Sun-cured tobacco	德阳市烟草公司 Deyang Tobacco Company	中国广东 Guangdong, China
208	城口晒烟 Chengkoushaiyan	晒烟 Sun-cured tobacco	德阳市烟草公司 Deyang Tobacco Company	中国重庆 Chongqing, China
209	4	晒烟 Sun-cured tobacco	德阳市烟草公司 Deyang Tobacco Company	中国四川 Sichuan, China
210	5	晒烟 Sun-cured tobacco	德阳市烟草公司 Deyang Tobacco Company	中国四川 Sichuan, China
211	良种 Liangzhong	晒烟 Sun-cured tobacco	德阳市烟草公司 Deyang Tobacco Company	中国湖南 Hunan, China
212	辅善 Fushan	晒烟 Sun-cured tobacco	德阳市烟草公司 Deyang Tobacco Company	中国四川 Sichuan, China
213	满耳朵 Manerduo	晒烟 Sun-cured tobacco	德阳市烟草公司 Deyang Tobacco Company	中国四川 Sichuan, China
214	青梗 Qinggeng	晒烟 Sun-cured tobacco	德阳市烟草公司 Deyang Tobacco Company	中国广东 Guangdong, China
215	南雄晒黄烟 Nanxiongshaihuangyan	晒烟 Sun-cured tobacco	德阳市烟草公司 Deyang Tobacco Company	中国广东 Guangdong, China
216	泉烟 Quanyan	晒烟 Sun-cured tobacco	德阳市烟草公司 Deyang Tobacco Company	中国四川 Sichuan, China
217	Maryland609	马里兰烟 Maryland tobacco	德阳市烟草公司 Deyang Tobacco Company	美国 USA
218	TN86	白肋烟 Burley	德阳市烟草公司 Deyang Tobacco Company	美国 USA
219	81-26	晒烟 Sun-cured tobacco	德阳市烟草公司 Deyang Tobacco Company	中国黑龙江 Heilongjiang, China
220	武鸣牛利 Wumingniuli	晾烟 Air-cured tobacco	国家烟草种质资源中期库 National Infrastructure for Crop Germplasm Resources (Tobacco, Qingdao)	中国广西 Guangxi, China

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参考文献原文顺序
文献年度倒序
文中引用次数倒序
被引期刊影响因子

[1]

刘国顺. 烟草栽培学. 北京: 中国农业出版社, 2003. pp 21-22.

[本文引用: 1]

Liu G

. Tobacco Cultivation. Beijing: China Agriculture Press, 2003. pp 21-22(in Chinese).

[本文引用: 1]

[2]

童治军. 烟草微卫星标记的开发与应用
浙江大学博士学位论文, 浙江杭州, 2012.

[本文引用: 1]

Tong Z

. Development and Application of Microsatellite Markers in Flue-cured Tobacco (Nicotiana tabacum L.)
PhD Dissertation of Zhejiang University, Hangzhou, Zhejiang, China, 2012 (in Chinese with English abstract).

[本文引用: 1]

[3]

任民, 王志德, 牟建民, 刘艳华, 张兴伟. 我国烟草种质资源的种类与分布概况
中国烟草科学, 2009,30(增刊1):8-14.

[本文引用: 1]

Ren

, Wang Z

, Mu J

, Liu Y

, Zhang X

. Overview of species and distribution of tobacco germplasm resources in China
Chin Tob Sci, 2009,30(S1):8-14 (in Chinese with English abstract).

[本文引用: 1]

[4]

杨柳, 汪斌, 童治军, 黄勇兵, 曹梅秀, 吴海乔, 巫升鑫, 陈顺辉, 兰涛. 25份普通烟草种质资源遗传多样性的SSR标记分析
福建农林大学学报(自然科学版), 2013,42(2):171-175.

[本文引用: 2]

Yang

, Wang

, Tong Z

, Huang Y

, Cao M

, Wu H

, Wu S

, Chen S

, Lan

. Genetic diversity analysis of 25 Nicotiana tabacum L. germplasm resources based on SSR marker
J Fujian Agric For Univ (Nat Sci Edn), 2013,42:171-175 (in Chinese with English abstract).

[本文引用: 2]

[5]

尹国英. 烟草种质资源遗传多样性和应用潜力分析以及核心SSR引物筛选
西南大学硕士学位论文, 重庆, 2013.

[本文引用: 1]

Yin G

. Analysis of Genetic Diversity and Application Potential of Tobacco Germplasm Resources and Screening of Core SSR Primers
MS Thesis of Southwest University, Chongqing, China, 2013 (in Chinese with English abstract).

[本文引用: 1]

[6]

李爱军, 秦艳青, 代惠娟, 范静苑, 曾代龙, 王华敬, 潘运金. 国产雪茄烟叶科学发展刍议
中国烟草学报, 2012,18(1):112-114.

[本文引用: 1]

Li A

, Qin Y

, Dai H

, Fan J

, Zeng D

, Wang H

, Pan Y

. On scientific development of China’s cigar leaf
Acta Tab Sin, 2012,18(1):112-114 (in Chinese with English abstract).

[本文引用: 1]

[7]

Piano L

, Abet

, Sorrentino

, Acanfora

, Cozzolino

, Muro A

. Genetic variability in Nicotiana tabacum and Nicotiana species as revealed by RAPD markers: 1. Development of the RAPD procedure
Nephron Clin Prac, 2000,19:1-15.

[本文引用: 1]

[8]

常爱霞, 瞿永生, 贾兴华. 烟草RAPD反应体系优化及品种多态性标记研究
中国烟草科学, 2004,25(2):9-13.

[本文引用: 1]

Chang A

, Qu Y

, Jia X

. Optimalization of tobacco RAPD reaction system and studies on polymorphic marker of tobacco varieties
Chin Tob Sci, 2004,25(2):9-13 (in Chinese with English abstract).

[本文引用: 1]

[9]

Arslan

, Okumus

. Genetic and geographic polymorphism of cultivated tobaccos (Nicotiana tabacum) in Turkey
Genetika, 2006,42:818-823.

URLPMID:16871787 [本文引用: 1]

Nicotiana tabacum (2n = 48) is a natural amphidiploid and shows a distribution over a geographical area in eastern anatolia. Random amplified polymorphic DNA (RAPD) technique was used to evaluate both genetic diversity among 21 primitive tobacco accessions comparing flue cure virginia genotype (FCV) and their geographical polymorphism as a source of genetic variations for breeding programs. Only 13 of all the 60 random primers used in RAPD showed polymorphism acceptable for characterization of these accessions. Totally 118 RAPD fragments were generated from thirteen decamer primer and sixtyfour of them were found polymorphic (54.2%). Mus and FCV showed the smallest genetic distance among accessions cultivated in the eastern anatolia. These results shows that the RAPD assay is a powerful approach for identifying genetic and geographic polymorphism.

[10]

Dadras A

, Sabouri

, Nejad G

, Sabouri

, Shoai-Deylami

. Association analysis, genetic diversity and structure analysis of tobacco based on AFLP markers
Mol Biol Rep, 2014,41:3317-3329.

[本文引用: 1]

[11]

杨友才, 周清明, 尹晗琪, 朱列书. 烟草种质资源遗传多样性及亲缘关系的AFLP分析
中国农业科学, 2006,39:2194-2199.

[本文引用: 1]

Yang Y

, Zhou Q

, Yin H

, Zhu L

. Studies on genetic diversity and relationship in tobacco germplasms by AFLP analysis
Sci Agric Sin, 2006,39:2194-2199 (in Chinese with English abstract).

[本文引用: 1]

[12]

Ren

, Timko M

. AFLP analysis of genetic polymorphism and evolutionary relationships among cultivated and wild Nicotiana species
Genome, 2001,44:559-571.

URLPMID:11550889 [本文引用: 1]

Amplified fragment length polymorphism (AFLP) analysis was used to determine the degree of intra- and inter-specific genetic variation in the genus Nicotiana. Forty-six lines of cultivated tobacco (Nicotiana tabacum L.) and seven wild Nicotiana species, including N. sylvestris, N. tomentosiformis, N. otophora, N. glutinosa, N. suaveolens, N. rustica, and N. longiflora, were analyzed, using at least eight different oligonucleotide primer combinations capable of detecting a minimum of 50 polymorphic bands per primer pair. The amount of genetic polymorphism present among cultivated tobacco lines (N. tabacum) was limited, as evidenced by the high degree of similarity in the AFLP profiles of cultivars collected worldwide. Six major clusters were found within cultivated tobacco that were primarily based upon geographic origin and manufacturing quality traits. A greater amount of genetic polymorphism exists among wild species of Nicotiana than among cultivated forms. Pairwise comparisons of the AFLP profiles of wild and cultivated Nicotiana species show that polymorphic bands present in N. tabacum can be found in at least one of three proposed wild progenitor species (i.e., N. sylvestris, N. tomentosiformis, and N. otophora). This observation provides additional support for these species contributing to the origin of N. tabacum.

[13]

龙腾, 刘雷, 黄玉碧. 四川部分晾晒烟种质遗传关系的SRAP分析
作物学报, 2009,35:173-178.

[本文引用: 1]

Teng

, Liu

, Huang Y

. Genetic analvsis of Sichuan sun-cured tobacco germplasm by SRAP
Acta Agron Sin, 2009,35:173-178.

[本文引用: 1]

[14]

张吉顺, 王仁刚, 杨春元, 吴春, 史跃伟, 王志红, 王轶, 任学良. 国内外烤烟品种农艺性状的遗传多样性及与SRAP标记的关联分析
作物学报, 2012,38:1029-1041.

[本文引用: 1]

Zhang J

, Wang R

, Yang C

, Wu

, Shi Y

, Wang Z

, Wang

, Ren X

. Genetic diversity of agronomic traits and association analysis with SRAP markers in flue-cured tobacco ( Nicotiana tabacum) varieties from China and abroad
Acta Agron Sin, 2012,38:1029-1041 (in Chinese with English abstract).

[本文引用: 1]

[15]

Qiong

, Renxiang

, Wei

. Fingerprint map construction of core tobacco germplasm by ISSR markers
Southwest China J Agric Sci, 2010,23:335-339.

[本文引用: 1]

[16]

Edrisi M

, Samizadeh L

, Shoaei D

. Assessing the genetic diversity of tobacco ( Nicotiana tabacum L.) varieties
Crop Breed J, 2012,2:125-132.

[本文引用: 1]

[17]

梁景霞, 祁建民, 方平平, 王涛, 陈顺辉, 周东新, 陶爱芬, 梁康迳, 吴为人. 烟草种质资源遗传多样性与亲缘关系的ISSR聚类分析
中国农业科学, 2008,41:286-294.

[本文引用: 1]

Liang J

, Qi J

, Fang P

, Wang

, Chen S

, Zhou D

, Tao A

, Liang K

, Wu W

. Genetic diversity and genetic relatives analysis of tobacco germplasm based on inter-simple sequence repeat (ISSR)
Sci Agric Sin, 2008,41:286-294 (in Chinese with English abstract).

[本文引用: 1]

[18]

Salazar J

, Rubio

, Ruiz

, Tartarini

, Gómez P

, Dondini

. SNP development for genetic diversity analysis in apricot
Tree Genet Genomes, 2015,11:15.

[本文引用: 2]

[19]

Wang W

, Mauleon

, Hu Z

, Chebotarov

, Tai S

, Wu Z

, Li

, Zheng T

, Fuentes R

, Zhang

, Mansueto

, Copetti

, Sanciangco

, Palis K

, Xu J

, Sun

, Fu B

, Zhang H

, Gao Y

, Zhao X

, Shen

, Cui

, Yu

, Li Z

, Chen M

, Detras

, Zhou Y

, Zhang X

, Zhao

, Kudrna

, Wang C

, Li

, Jia

, Lu J

, He X

, Dong Z

, Xu J

, Li Y

, Wang

, Shi J

, Li

, Zhang D

, Lee

, Hu W

, Poliakov

, Dubchak

, Ulat V

, Borja F

, Mendoza J

, Ali

, Li

, Gao

, Niu Y

, Yue

, Naredo

M E B

, Talag

, Wang

X Q

, Li

J J

, Fang

X D

, Yin

, Glaszmann

J C

, Zhang

J W

, Li

J Y

, Hamilton

R S

, Wing

R A

, Ruan

, Zhang

G Y

, Wei

C C

, Alexandrov

, McNally

K L

, Li

Z K

, Leung

. Genomic variation in 3,010 diverse accessions of Asian cultivated rice
Nature, 2018,557:43-49.

[本文引用: 2]

[20]

尚维, 赵申清玉, 党江波, 郭启高, 梁国鲁, 杨超, 张艳, 陈益银. 基于SSR分子标记的Nicotiana tobacum-N. plumbaginifolia异源染色体植株的鉴定与筛选
作物学报, 2018,44:1640-1649.

[本文引用: 2]

Shang

, Zhao

S Q Y

, Dang

J B

, Guo

Q G

, Liang

G L

, Yang

, Zhang

, Chen

Y Y

. Identification and screening of
Nicotiana tobacam-N. plumbaginifolia heterologous chromosome plants based on SSR marker. Acta Agron Sin, 2018,44:1640-1649 (in Chinese with English abstract).

[本文引用: 2]

[21]

焦荻, 商纪鹏, 高素燕, 王钦, 郝建全, 何伟, 焦定量, 吕敬刚, 陈则明. 西瓜品种‘蜜多’种子纯度SSR标记鉴定
中国瓜菜, 2019,32(7):19-22.

[本文引用: 1]

Jiao

, Shang J

, Gao S

, Wang

, Hao J

, He

, Jiao D

, Lyu J

, Chen Z

. Identification of ‘Miduo’ watermelon seed purity using SSR marker
China Cucurbits Veget, 2019,32(7):19-22 (in Chinese with English abstract).

[本文引用: 1]

[22]

卢媛, 艾为大, 韩晴, 王义发, 李宏杨, 瞿玉玑, 施标, 沈雪芳. 糯玉米自交系SSR标记遗传多样性及群体遗传结构分析
作物学报, 2019,45:214-224.

[本文引用: 2]

, Ai W

, Han

, Wang Y

, Li H

, Qu Y

, Shi

, Shen X

. Genetic diversity and population structure analysis by SSR markers in waxy maize
Acta Agron Sin, 2019,45:214-224 (in Chinese with English abstract).

[本文引用: 2]

[23]

赵靓, 罗燕杰, 肖思文, 蔡亚南, 何立飞, 遆羽静, 李庆卫. 基于表型和SSR标记的梅花种质资源遗传多样性分析
分子植物育种, 2019,17:4458-4469.

[本文引用: 1]

Zhao

, Luo Y

, Xiao S

, Cai Y

, He L

, Di Y

, Li Q

. Genetic diversity analysis on Prunus mume germplasm resources based on phenotypic traits and SSR markers
Mol Plant Breed, 2019,17:4458-4469 (in Chinese with English abstract).

[本文引用: 1]

[24]

张跃新, 胡伟, 郝艺铭, 闫宝松, 马凤, 么宏伟. 北方地区黑木耳部分主栽品种与野生菌株遗传多样性SSR分析
中国食用菌, 2019,38(5):44-48.

[本文引用: 1]

Zhang Y

, Hu

, Hao Y

, Yan B

, Ma

, Yao H

. Analysis on genetic diversity for the part of main cultivars and wild strains of auricularia auricula-judae from Northern China by SSR markers
Edible Fungi China, 2019,38(5):44-48 (in Chinese with English abstract).

[本文引用: 1]

[25]

倪维晨, 李瑞霞, 陶启威, 张戟, 毕研飞, 钱春桃. 基于SSR标记的地方品种糯性小玉米自交系指纹图谱构建
浙江农业科学, 2019,60:911-914.

[本文引用: 1]

Ni W

, Li R

, Tao Q

, Zhang

, Bi Y

, Qian C

. Construction of fingerprints of waxy maize inbred lines based on SSR markers
Zhejiang Agric Sci, 2019,60:911-914 (in Chinese with English abstract).

[本文引用: 1]

[26]

马斌, 李军乔, 刘贺贺, 刘欣, 李风英, 富贵, 白世俊, 韦梅琴. 蕨麻品种SSR指纹图谱的构建及遗传相似性分析
分子植物育种, 2019,17:4367-4377.

[本文引用: 1]

, Li J

, Liu H

, Liu

, Li F

, Fu

, Bai S

, Wei M

. Construction and analysis of genetic similarity of SSR fingerprints in Juema
Mol Plant Breed, 2019,17:4367-4377 (in Chinese with English abstract).

[本文引用: 1]

[27]

Bindler

, Hoeven

R V D

, Gunduz

, Plieske

, Ganal

, Rossi

, Gadani

, Donini

. A microsatellite marker based linkage map of tobacco
Theor Appl Genet, 2007,114:341-349.

DOI:10.1007/s00122-006-0437-5 URLPMID:17115128 [本文引用: 1]

We report the first linkage map of tobacco (Nicotiana tabacum L.) generated through microsatellite markers. The microsatellite markers were predominantly derived from genomic sequences of the Tobacco Genome Initiative (TGI) through bioinformatics screening for microsatellite motives. A total of 684 primer pairs were screened for functionality in a panel of 16 tobacco lines. Of those, 637 primer pairs were functional. Potential parents for mapping populations were evaluated for their polymorphism level through genetic similarity analysis. The similarity analysis revealed that the known groups of tobacco varieties (Burley, Flue-cured, Oriental and Dark) form distinct clusters. A mapping population, based on a cross between varieties Hicks Broad Leaf and Red Russian, and consisting of 186 F2 individuals, was selected for mapping. A total of 282 functional microsatellite markers were polymorphic in this population and 293 loci could be mapped together with the morphological trait flower color. Twenty-four tentative linkage groups spanning 1,920 cM could be identified. This map will provide the basis for the genetic mapping of traits in tobacco and for further analyses of the tobacco genome.

[28]

Bindler

, Plieske

, Bakaher

, Gunduz

, Ivanov

, Hoeven

R V D

, Ganal

, Donini

. A high density genetic map of tobacco (Nicotiana tabacum L.) obtained from large scale microsatellite marker development
Theor Appl Genet, 2011,123:219-230.

URLPMID:21461649 [本文引用: 2]

[29]

Lewis R

, Nicholson J

, Moon H

. Use of transferable Nicotiana tabacum L. microsatellite markers for investigating genetic diversity in the genus Nicotiana
Genome, 2008,51:547-559.

DOI:10.1139/G08-039 URLPMID:18650945 [本文引用: 1]

The recent development of microsatellite markers for tobacco, Nicotiana tabacum L., may be valuable for genetic studies within the genus Nicotiana. The first objective was to evaluate transferability of 100 N. tabacum microsatellite primer combinations to 5 diploid species closely related to tobacco. The number of primer combinations that amplified scorable bands in these species ranged from 42 to 56. Additional objectives were to assess levels of genetic diversity amongst available accessions of diploid relatives closely related to tobacco (species of sections Sylvestres and Tomentosae), and to evaluate the efficacy of microsatellite markers for establishing species relationships in comparison with existing phylogenetic reconstructions. A subset of 46 primer combinations was therefore used to genotype 3 synthetic tobaccos and an expanded collection of 51 Nicotiana accessions representing 15 species. The average genetic similarity for 7 diverse accessions of tobacco was greater than the average similarity for N. otophora accessions, but lower than the average genetic similarities for N. sylvestris, N. tomentosa, N. kawakamii, and N. tomentosiformis accessions. A microsatellite-based phylogenetic tree was largely congruent with taxonomic representations based on morphological, cytological, and molecular observations. Results will be useful for selection of parents for creation of diploid mapping populations and for germplasm introgression activities.

[30]

Moon H

, Nicholson J

, Heineman

, Lion

, Hoeven

R V D

, Hayes

A J

, Lewis

R S

. Changes in genetic diversity of U.S. flue-cured tobacco germplasm over seven decades of cultivar development
Crop Sci, 2009,49:498-508.

[本文引用: 1]

[31]

Moon H

, Nifong J

, Nicholson J

, Heineman

, Lion

, Hoeven

R V D

, Hayes

A J

, Lewis

R S

. Microsatellite-based analysis of tobacco ( Nicotiana tabacum L.) genetic resources
Crop Sci, 2009,49:2149-2159.

[本文引用: 1]

[32]

马冰, 代帅帅, 程亚增, 蒋彩虹, 任民, 程立锐, 杨爱国. 烤烟种质资源SSR核心引物的筛选及验证
中国烟草科学, 2016,37(5):1-5.

[本文引用: 1]

, Dai S

, Cheng Y

, Jiang C

, Ren

, Cheng L

, Yang A

. Screen and identification of SSR core primers for flue-cured tobacco germplasm
Chin Tob Sci, 2016,37(5):1-5 (in Chinese with English abstract).

[本文引用: 1]

[33]

张雪廷, 童治军, 焦芳婵, 肖炳光, 曾千春. 38份晾晒烟种质资源遗传关系的SSR分析
植物遗传资源学报, 2013,14:653-658.

[本文引用: 1]

Zhang X

, Tong Z

, Jiao F

, Xiao B

, Zeng Q

. Genetic relationship analysis of thirty-eight sun/air-cured tobacco germplasms based on simple sequence repeat (SSR) markers
J Plant Genet Resour, 2013,14(4):653-658 (in Chinese with English abstract).

[本文引用: 1]

[34]

童治军, 陈学军, 方敦煌, 曾建敏, 吴兴富, 肖炳光. 231份烤烟种质资源SSR标记遗传多样性及其与农艺性状和化学成分的关联分析
中国烟草学报, 2017,23(5):31-40.

[本文引用: 3]

Tong Z

, Chen X

, Fang D

, Zeng J

, Wu X

, Xiao B

. SSR marker-based analyses on genetic diversity and relevant variations of agronomic traits and chemical composition of 231 flue-cured tobacco germplasm resources
Acta Tab Sin, 2017,23(5):31-40 (in Chinese with English abstract).

[本文引用: 3]

[35]

刘国祥, 邹昆晏, 任民, 佟英, 冯全福, 杨爱国, 戴培刚, 徐宜民, 张兴伟. 33份晒烟种质资源SSR标记的指纹图谱构建
中国烟草学报, 2017,23(5):84-91.

[本文引用: 3]

Liu G

, Zou K

, Ren

, Tong

, Feng Q

, Yang A

, Dai P

, Xu Y

, Zhang X

. Construction of SSR markers’ fingerprints of 33 sun-cured tobacco germplasm resources
Acta Tab Sin, 2017,23(5):84-91 (in Chinese with English abstract).

[本文引用: 3]

[36]

张铭真, 李晓辉, 王袁, 杨铁钊, 徐世晓. 81份烟草种质资源遗传多样性分析
江西农业学报, 2017,29(1):62-68.

[本文引用: 1]

Zhang M

, Li X

, Wang

, Yang T

, Xu S

. Analysis of genetic diversity of 81 tobacco germplasm resources
Acta Agric Jiangxi, 2017,29(1):62-68 (in Chinese with English abstract).

[本文引用: 1]

[37]

陈芳, 徐世晓, 李晓辉, 刘超, 周建飞, 王袁, 田培, 杨铁钊. 基于SSR标记的80份烟草种质指纹图谱的构建及遗传多样性分析
作物杂志, 2019, ( 1):28-37.

[本文引用: 2]

Chen

, Xu S

, Li X

, Liu

, Zhou J

, Wang

, Tian

, Yang T

. Construction of molecular fingerprinting and analysis of genetic diversity for 80 tobacco ( Nicotiana tabacum) germplasms based on SSR markers
Crops, 2019, ( 1):28-37 (in Chinese with English abstract).

[本文引用: 2]

[38]

樊文强, 盖红梅, 孙鑫, 杨爱国, 张忠锋, 任民. SSR数据格式转换软件DataFormater
分子植物育种, 2016,14:265-270.

[本文引用: 1]

Fan W

, Ge H

, Sun

, Yang A

, Zhang Z

, Ren

. DataFormater, a software for SSR data formatting to develop population genetics analysis
Mol Plant Breed, 2016,14:265-270 (in Chinese with English abstract).

[本文引用: 1]

[39]

Yen

, Boyle

. Population genetic analysis of co-dominant and dominant markers and quantitative traits
Belg J Bot, 1997,129:157.

[本文引用: 1]

[40]

Sándor

, Péter

, István

, Mousapour G

, Géza

, János

. PICcalc: an online program to calculate polymorphic information content for molecular genetic studies
Biochem Genet, 2012,50:670-672.

DOI:10.1007/s10528-012-9509-1 URLPMID:22573137 [本文引用: 1]

[41]

Sudhir

, Glen

, Koichiro

. MEGA7: molecular evolutionary genetics analysis Version 7.0 for bigger datasets
Mol Biol Evol, 2016,33:1870-1874.

DOI:10.1093/molbev/msw054 URLPMID:27004904 [本文引用: 1]

We present the latest version of the Molecular Evolutionary Genetics Analysis (Mega) software, which contains many sophisticated methods and tools for phylogenomics and phylomedicine. In this major upgrade, Mega has been optimized for use on 64-bit computing systems for analyzing larger datasets. Researchers can now explore and analyze tens of thousands of sequences in Mega The new version also provides an advanced wizard for building timetrees and includes a new functionality to automatically predict gene duplication events in gene family trees. The 64-bit Mega is made available in two interfaces: graphical and command line. The graphical user interface (GUI) is a native Microsoft Windows application that can also be used on Mac OS X. The command line Mega is available as native applications for Windows, Linux, and Mac OS X. They are intended for use in high-throughput and scripted analysis. Both versions are available from www.megasoftware.net free of charge.

[42]

Pritchard J

, Stephens

, Donnelly

. Inference of population structure using multilocus genotype data
Genetics, 2000,155:945-959.

URLPMID:10835412 [本文引用: 1]

We describe a model-based clustering method for using multilocus genotype data to infer population structure and assign individuals to populations. We assume a model in which there are K populations (where K may be unknown), each of which is characterized by a set of allele frequencies at each locus. Individuals in the sample are assigned (probabilistically) to populations, or jointly to two or more populations if their genotypes indicate that they are admixed. Our model does not assume a particular mutation process, and it can be applied to most of the commonly used genetic markers, provided that they are not closely linked. Applications of our method include demonstrating the presence of population structure, assigning individuals to populations, studying hybrid zones, and identifying migrants and admixed individuals. We show that the method can produce highly accurate assignments using modest numbers of loci-e.g. , seven microsatellite loci in an example using genotype data from an endangered bird species. The software used for this article is available from http://www.stats.ox.ac.uk/ approximately pritch/home. html.

[43]

Evanno

, Regnaut

, Goudet

. Detecting the number of clusters of individuals using the software STRUCTURE: a simulation study
Mol Ecol, 2005,14:2611-2620.

DOI:10.1111/j.1365-294X.2005.02553.x URLPMID:15969739 [本文引用: 2]

The identification of genetically homogeneous groups of individuals is a long standing issue in population genetics. A recent Bayesian algorithm implemented in the software STRUCTURE allows the identification of such groups. However, the ability of this algorithm to detect the true number of clusters (K) in a sample of individuals when patterns of dispersal among populations are not homogeneous has not been tested. The goal of this study is to carry out such tests, using various dispersal scenarios from data generated with an individual-based model. We found that in most cases the estimated 'log probability of data' does not provide a correct estimation of the number of clusters, K. However, using an ad hoc statistic DeltaK based on the rate of change in the log probability of data between successive K values, we found that STRUCTURE accurately detects the uppermost hierarchical level of structure for the scenarios we tested. As might be expected, the results are sensitive to the type of genetic marker used (AFLP vs. microsatellite), the number of loci scored, the number of populations sampled, and the number of individuals typed in each sample.

[44]

王静毅, 陈业渊, 黄秉智, 于飞, 武耀廷. 部分香蕉品种SSR 指纹图谱的构建
果树学报, 2009,26:733-738.

[本文引用: 1]

Wang J

, Chen Y

, Huang B

, Yu

, Wu Y

. Establishment of fingerprinting for bananas (Musa nana) by SSR marker
J Fruit Sci, 2009,26:733-738 (in Chinese with English abstract).

[本文引用: 1]

[45]

宋海斌, 崔喜波, 马鸿艳, 朱子成, 栾非时. 基于SSR 标记的甜瓜品种(系)DNA 指纹图谱库的构建
中国农业科学, 2012,45:2676-2689.

[本文引用: 1]

Song H

, Cui X

, Ma H

, Zhu Z

, Luan F

. Construction of DNA fingerprint database based on SSR marker for varieties (lines) of Cucumis melo L
Sci Agric Sin, 2012,45:2676-2689 (in Chinese with English abstract).

[本文引用: 1]

[46]

陈越, 陈玲, 李春花, 张敦宇, 付坚, 程在全. 中国南方地区水稻资源SSR指纹数据库的构建及遗传多样性分析
分子植物育种, 2020,18:6502-6517.

[本文引用: 1]

Chen

, Chen

, Li C

, Zhang D

, Fu

, Cheng Z

. Construction of SSR fingerprint database and genetic diversity analysis of rice resources in Southern China
Mol Plant Breed, 2020,18:6502-6517 (in Chinese with English abstract).

[本文引用: 1]

[47]

苏一钧, 王娇, 戴习彬, 唐君, 赵冬兰, 张安, 周志林, 曹清河. 303份甘薯地方种SSR遗传多样性与群体结构分析
植物遗传资源学报, 2018,19:243-251.

[本文引用: 1]

Su Y

, Wang

, Dai X

, Tang

, Zhao D

, Zhang

, Zhou Z

, Cao Q

. Genetic diversity and population structure analysis of 303 sweet potato landraces using SSR markers
J Plant Genet Resour, 2018,19:243-251 (in Chinese with English abstract).

[本文引用: 1]

[48]

许美玲, 贺晓辉, 宋玉川, 刘勇, 李梅云, 樊有银, 李永平. 72份雪茄烟种质资源的鉴定评价和聚类分析
中国烟草学报, 2017,23(5):41-56.

[本文引用: 1]

Xu M

, He X

, Song Y

, Liu

, Li M

, Fan Y

, Li Y

. Identification, evaluation and cluster analysis of 72 cigar tobacco germplasm resources
Acta Tab Sin, 2017,23(5):41-56 (in Chinese with English abstract).

[本文引用: 1]

[49]

许美玲, 贺晓辉, 宋玉川, 李梅云, 方敦煌, 陈学军, 樊有银, 李永平. 76份雪茄烟资源鉴定评价
中国烟草学报, 2018,24(5):18-26.

[本文引用: 1]

Xu M

, He X

, Song Y

, Li M

, Fang D

, Chen X

, Fan Y

, Li Y

. Identification and comprehensive evaluation of 76 germplasms of cigar tobacco
Acta Tab Sin, 2018,24(5):18-26 (in Chinese with English abstract).

[本文引用: 1]

[50]

陈杰, 杨静, 龙胜贤, 肖慈平, 杨昌义, 黄清忠, 王维. SSR分子标记在烟草研究中的应用进展
生物技术通报, 2015,31(3):43-48.

[本文引用: 1]

Chen

, Yang

, Long S

, Xiao C

, Yang C

, Huang Q

, Wang

. Advance of the application of SSR molecular markers in tobacco research
Biotechnol Bull, 2015,31(3):43-48 (in Chinese with English abstract).

[本文引用: 1]

[51]

赖瑞强, 李荣华, 夏岩石, 郭培国, 袁清华, 赵伟才. 烟草种质的SSR标记遗传多样性及青枯病抗性的关联分析
中国烟草学报, 2018,24(6):71-81.

[本文引用: 1]

Lai R

, Li R

, Xia Y

, Guo P

, Yuan Q

, Zhao W

. SSR marker-based genetic diversity analysis of tobacco germplasm and association analysis with resistance to bacterial wilt
Acta Tab Sin, 2018,24(6):71-81 (in Chinese with English abstract).

[本文引用: 1]

[52]

丛鑫, 刘艳华, 戴培刚, 王志德, 张兴伟, 任民, 程立锐, 姚志敏. 抗PVY烟草种质的遗传多样性分析
植物遗传资源学报, 2014,15:679-684.

[本文引用: 1]

Cong

, Liu Y

, Dai P

, Wang Z

, Zhang X

, Ren

, Cheng L

, Yao Z

. Genetic diversity analysis of tobacco germplasm resistance to PVY
J Plant Genet Resour, 2014,15:679-684 (in Chinese with English abstract).

[本文引用: 1]

[53]

孙九喆, 杨金初, 苏东嬴, 王二彬, 王君婷, 张顺峰. 基于SSR标记的初烤烟叶品种快速鉴别
烟草科技, 2019,52(3):32-38.

[本文引用: 1]

Sun J

, Yang J

, Su D

, Wang E

, Wang J

, Zhang S

. Rapid identification of cured tobacco leaf varieties based on SSR markers
Tob Sci Technol, 2019,52(3):32-38 (in Chinese with English abstract).

[本文引用: 1]

[54]

Hilde

, Kurt

, Bjorn

. DNA fingerprinting in botany: past, present, future
Invest Genet, 2014,5:1.

[本文引用: 1]

[55]

王忠华. DNA指纹图谱技术及其在作物品种资源中的应用
分子植物育种, 2006,4:425-430.

[本文引用: 1]

Wang Z

. DNA Fingerprinting technology and its application in crop germplasm resources
Mol Plant Breed, 2006,4:425-430 (in Chinese with English abstract).

[本文引用: 1]

[56]

徐军. 烟草核心种质SSR指纹图谱构建及遗传多样性分析
中国农业科学院研究生院硕士学位论文, 山东青岛, 2011.

[本文引用: 1]

. Construction of Fingerprinting and Analysis of Genetic Diversity with SSR Marker for Tobacco Core Germplasm
MS Thesis of Chinese Academy of Agricultural Sciences, Qingdao, Shandong, China, 2011 (in Chinese with English abstract).

[本文引用: 1]