古蔺牛皮茶种质资源遗传多态性

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刘绍杰, 迟琳, 谢文钢, 韩楠, 陈应会, 何鑫, 唐茜. 古蔺牛皮茶种质资源遗传多态性. 作物学报, 40(12): 2118-2127
LIU Shao-Jie, CHI Lin, XIE Wen-Gang, HAN Nan, CHEN Ying-Hui, HE Xin, TANG Qian. Genetic Diversity of Gulin-niupicha [Camellia sinensis (L.)O. Kuntze] Germplasm- Resources . Acta Agronomica Sinica, 40(12): 2118-2127 复制到剪切板

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作物学报编辑部

古蔺牛皮茶种质资源遗传多态性

刘绍杰^1,², 迟琳², 谢文钢¹, 韩楠¹, 陈应会², 何鑫², 唐茜^1,^*

¹四川农业大学园艺学院, 四川雅安 625014

²重庆市经贸中等专业学校农林水利系, 重庆 402100

^*通讯作者(Corresponding author): 唐茜, E-mail:tangqi2008@126.com

收稿日期:2014-09-16

基金:本研究由四川省科技支撑计划项目(2014NZ0105)资助;

摘要

古蔺牛皮茶是四川省特有的茶树种质资源, 原产于四川省泸州市古蔺县古蔺镇枣林村椒子沟。为了更好地保护和利用珍稀的茶树种质资源, 在椒子沟山区选取7个居群共76株典型的古蔺牛皮茶树, 采用实地调查记录、生化成分及ISSR分子标记检测等方法, 分析该种质资源的形态学特征、品质生化成分及遗传多态性。结果表明: (1)古蔺牛皮茶树型为灌木或小乔木; 成熟叶片为绿色或深绿色, 主要为中叶, 叶形为椭圆或长椭圆; 新梢芽叶玉白、黄绿、绿色或紫绿色。表明古蔺牛皮茶居群间表型特征多样化。(2)在7个居群中鉴定出高茶多酚(春梢>27%, 夏梢>31%)和儿茶素(春梢>150 mg g^-1, 夏梢>190 mg g^-1)的居群2个, 游离氨基酸较高( >4.5%)的居群2个; 7个居群的酚氨比值在3.84~16.95之间。表明古蔺牛皮茶在茶树育种和茶叶加工等方面具有较高的利用价值。(3) 15条ISSR引物共扩增出135条谱带, 其中122条为多态性条带, 多态性位点百分比为90.4%。供试材料间的平均Nei’s基因多态性指数(H)和Shannon’s信息指数(I)分别为0.254和0.392, 表明材料间的遗传多态性较低。

关键词: 古蔺牛皮茶; 种质资源; 遗传多态性; ISSR

Genetic Diversity of Gulin-niupicha [Camellia sinensis (L.)O. Kuntze] Germplasm- Resources

LIU Shao-Jie^1,², CHI Lin², XIE Wen-Gang¹, HAN Nan¹, CHEN Ying-Hui², HE Xin², TANG Qian^1,^*

¹College of Horticulture, Sichuan Agricultural University, Ya#cod#x02019;an 625014, China

²Department of Agriculture, Forestry and Water Conservancy, Chongqing Economic and Trade Vocational School, Chongqing 402100, China

Abstract

Gulin-niupicha (Camellia sinensis) is a tea landrace population specifically grown in Jiaozigou, Gulin County, Sichuan Province, China. To protect and utilize Gulin-niupicha resources effectively, we investigated the morphologic characteristics, biochemical components, and genetic diversity based on inter-simple sequence repeat (ISSR) markers by sampling 76 individuals from seven populations of Gulin-niupicha germplasm. The results showed that: (1) there were various biological characteristics among the seven populations. Importantly, a majority of Gulin-niupicha were shrubs or semi-arbor. Their mature leaves were medium in size, elliptic or oblong in shape, and in green or dark green color; while the young shoots showed more colors varying form whitish, yellow green, green or purple green. (2) In the seven populations, two had high contents of free amino acids (>4.5%) and two had both high contents of tea polyphenols (>27% in spring shoots and >31% in summer shoots) and catechins (>150 mg g^-1 in spring shoots and >190 mg g^-1 in summer shoots). The ratio of polyphenols-to-free amino acids ranged from 3.84 to 16.95 in the seven populations. The results of biochemical component measurements indicated that the Gulin-niupicha germplasm resources are of great value in tea tree breeding and tea processing. (3) A total of 135 fragments were amplified in the 76 accessions using 15 pairs of ISSR primers, in which 90.4% (122/135) were polymorphic bands. The genetic diversity of the Gulin-niupicha germplasm revealed by ISSR markers was low, with the Nei’s gene diversity index of 0.254 and Shannon’s information index of 0.392.

Keyword: Gulin-niupicha (Camellia sinensisL.); Germplasm resources; Genetic diversity; ISSR

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种质资源是在漫长的历史过程中, 由自然演化和人工选择形成的一种重要的自然资源。它蕴藏着各种性状的遗传基因, 是选育新品种和发展农业生产的物质基础, 也是进行生物学研究的重要材料^{[ 1]}。我国西南地区的云贵川一带是茶树的发源地之一, 拥有丰富的野生、半野生和栽培的茶树种质资源^{[ 2]}。古蔺牛皮茶是四川省的一个重要地方茶树品种资源, 为有性系品种, 主要分布在古蔺县古蔺镇枣林村的椒子沟, 1985年该品种被认定为四川省省级地方品种。古蔺牛皮茶生长和栽培区域仅限于椒子沟及周围的山区和丘陵地区, 其性状、品质特殊, 有独特的农艺价值。目前有关古蔺牛皮茶的研究较少^{[ 3]}, 未见对古蔺牛皮茶系统研究的报道; 同时, 该品种资源也未得到良好的利用和推广, 加上该地区连年来干旱少雨的气候条件和人为破坏, 导致古蔺牛皮茶群体的生长区域不断缩小, 种质资源流失, 所以应对古蔺牛皮茶种质资源的性状进行系统的鉴定和评价, 以加强对该资源的保护和利用。

本文通过调查古蔺牛皮茶群体的分布情况, 初步分析了不同居群茶树的形态学特征、品质生化成分及遗传多态性, 旨在探究古蔺牛皮茶的资源现状、农艺价值和遗传特征, 为开展古蔺牛皮茶种质资源的收集、保存、评价和利用以及特异资源的发掘和新品种的选育奠定基础。

1 材料与方法

1.1 茶树生长区域自然概况

古蔺县地处四川盆地南缘向贵州高原过度地带。其四季分明、雨热同季、夏季炎热、冬无严寒, 气温差异大, 无霜期长, 年降雨量偏少, 湿度适中, 日照较充足, 常年多夏伏旱, 具有四川盆地南部高温和贵州高原乍寒乍暖的气候特点。古蔺牛皮茶集中分布于该县古蔺镇枣林村的椒子沟(27°58′33″~ 27°59′30″N, 105°46′39″~105°47′33″E, 海拔900~1100 m的山林中), 距离古蔺县城5 km, 分布区域约20 km², 海拔800~1200 m。茶树多为野生、半野生状态, 也有少量人工栽培的茶树。

1.2 供试材料

从椒子沟集中分布的野生、半野生古蔺牛皮茶群体中, 选定7个数量较多、外部形态特征具有代表性的居群共计76株( 表1), 观测并记录其地理位置。以当地栽培的四川中小叶群体品种为对照组(CK)。

表1 古蔺牛皮茶种质资源来源情况 Table 1 Origin and sample number of Gulin-niupicha germplasm resources

1.3 形态学特征观察

于2012年春季驻点观测, 根据NY/T 1312-2007农作物种质资源鉴定技术规程(茶树)^{[ 4]}的方法, 观测古蔺牛皮茶树体、芽叶及成熟叶片的性状及特征, 并按相应方法和要求^{[ 1]}记录于茶树种质资源数据采集表。为方便后期的统计分析, 参照沈程文^{[ 5]}的方法量化处理形态学特征的质量指标( 表2)。

表2 古蔺牛皮茶种质资源表型性状编码 Table 2 Phenotypic traits and their coding for Gulin-niupicha germplasm resources

1.4 品质生化成分测定

于2012年春、夏两季及2013年春季采摘茶树的标准一芽二叶制作生化样, 根据NY/T 1312-2007农作物种质资源鉴定技术规程(茶树)^{[ 4]}制定的方法, 测定古蔺牛皮茶的品质生化成分, 测定指标包括茶多酚^{[ 7]}、游离氨基酸^{[ 8]}和咖啡碱^{[ 9]}。采用高效液相色谱法(HPLC)测定儿茶素和茶氨酸^{[ 7]}。

1.5 ISSR分析

于2012年春季取样, 选取茶树新梢一芽一叶, 迅速用液氮处理, 并将其保存在-80℃冰箱中备用。参照刘本英等^{[ 10]}的方法提取茶树基因组DNA。

参照刘本英等^{[ 10]}的ISSR反应体系, 20 μL反应体系中含模板DNA 30 ng, 10×缓冲液, Mg²⁺ 2.0 mmol L^-1, 引物0.2 μmol L^-1, dNTPs 0.4 mmol L^-1, Taq DNA聚合酶1.5 U。采用美国百乐公司生产的MyCycler T_m Thermal型PCR仪扩增DNA, 扩增程序为94℃预变性5 min; 94℃变性1 min, 48~62℃退火30 s, 72℃延伸2 min, 35次循环; 72℃再延伸10 min, 12℃保存。扩增产物在1×TBE缓冲液中, 用2%的琼脂糖凝胶电泳检测, 电压设定为90 V, 电泳3 h。经EB溶液染色后, 用SYNGENE (德国)凝胶成像系统拍照记录。

从加拿大哥伦比亚大学(University of British Columbia Biotechnology, UBS)报道的100条植物通用ISSR引物^{[ 11]}中挑选72条, 委托北京华大基因有限公司合成。参考引物的 T_m值, 采用梯度PCR确定退火温度。以古蔺牛皮茶N1为模板, 在48~62℃间设置0.5℃为一个温度梯度进行扩增, 扩增产物经1.5%琼脂糖凝胶电泳检测, 以扩增带型清楚杂带比较少的温度作为该引物的退火温度。选择能扩增出目的条带、多态性高、重复性好的引物作为资源扩增的核心引物。

将核心引物的扩增结果中清晰的条带赋值为1, 同一位置无带或难以分辨的弱带赋值为0, 建立原始数据矩阵。在同一扩增位点, 只要有至少一个材料在该扩增位点计数为“0”, 则该扩增位点为多态性条带, 每条多态性带可视为一个等位基因。

1.6 统计分析

用SPSS Version19.0软件统计分析形态学原始数据, 利用Duncan’s多重比较法进行差异显著性检验。参照罗晓莹等^{[ 12]}的方法, 利用POPGEN Version 1.32软件分析ISSR标记试验数据。多态位点百分率(PPL)、Nei’s基因多态性指数( H)、Shannon’s多态性信息指数( I)、各居群内基因遗传分化指数( G_st) ( G_st=1- H_s/ H_t)及居群内的遗传距离和相似系数等参数均由POPGEN Version 1.32软件获得。居群每代迁移数 N_m=(1- F_st)/4 F_st^{[ 13]}。将POPGEN Version 1.32软件得出的居群遗传相似系数输入NTsys 2.10e软件,得到居群的亲缘关系聚类图, 并测试形态学特征矩阵和ISSR分子标记数据矩阵的Mantel相关性。

2 结果与分析

2.1 茶树居群的形态学性状

7个供试的古蔺牛皮茶居群中, 树型大部分为灌木, 其中居群D和居群G全部为灌木型, 而其他5个居群均含有小乔木或乔木型。每个居群的平均树高在180.8~382.1 cm之间, 均比四川中小叶群体茶树(平均172.4 cm)高。其中, 居群B最高,为382.0 cm; 居群D最矮, 为180.0 cm。古蔺牛皮茶居群的成熟叶片特征呈多样化,叶片大小指数平均为1.54 (居群E)~2.50 (居群A), 大部分为2.0~2.5, 多为中叶种, 群体E的叶片大小指数平均为1.54, 含有部分小叶种茶树。叶形指数平均为1.18~1.40, 表明多数茶树的成熟叶片呈椭圆形,也有部分呈长椭圆形。其中,居群A、G的叶形指数分别为1.40、1.33, 含有较多的长椭圆叶形的茶树, 还含有少量披针叶形的茶树。叶色指标为1.60~2.09, 多数茶树叶片呈绿色, 少部分呈深绿色, 黄绿色的很少。各居群的芽叶性状差异也很大, 芽叶色泽从玉白色到紫绿色不等, 平均芽色指标在2.0~3.0之间, 其中, 居群A最低(2.1), 颜色相对偏浅, 而居群D (2.9)最高, 颜色相对较深。

2.2 品质生化成分分析

2.2.1 2012年春夏梢茶树采收的对象是新梢, 因此新梢中主要生化品质成分的含量和配比决定茶叶的先天品质。由表4可见, 7个古蔺牛皮茶居群新梢的茶多酚含量, 春梢为22.57%~30.17%, 夏梢为26.82%~36.57%。其中居群B (春梢为30.17%, 夏梢为36.57%)、F (春梢为28.97%, 夏梢为33.45%)和A (春梢为27.93%, 夏梢为31.51%)春、夏梢的含量均显著高于CK (春梢为25.60%, 夏梢为29.42%), 居群D春、夏梢均最低(春梢为22.57%, 夏梢为26.82%), 且显著低于CK。儿茶素含量, 春梢为126.52~157.15 mg g^-1, 夏梢为135.27~194.60 mg g^-1。其中居群A (春梢为157.15 mg g^-1, 夏梢为194.60 mg g^-1)、B (春梢为155.19 mg g^-1, 夏梢为191.69 mg g^-1)春、夏梢的含量均显著高于CK (148.07 mg g^-1、184.09 mg g^-1), 而居群C、F、G和D的春、夏梢含量则显著低于CK。游离氨基酸含量春梢为2.08%~4.84%, 夏梢为1.86%~ 3.97%, 除居群B和E显著低于CK (春梢为2.64%, 夏梢为2.33%)外, 其余居群春、夏梢均高于CK。其中居群D春、夏梢均最高(春梢为4.84%, 夏梢为3.97%), 居群C (春梢为4.63%, 夏梢为3.79%)次之。咖啡碱含量春梢为3.55%~4.81%, 夏梢为3.32%~ 5.22%, 除居群D和E外, 其他5个居群春梢和夏梢的含量均高于CK。酚氨比春梢为4.66~12.66, 夏梢为6.76~16.95, 其中居群E春、夏梢均最高, 居群B次之, 均显著高于CK, 居群D最低。

表3 古蔺牛皮茶居群间8个生物学表型性状统计分析 Table 3 Statistical analysis of eight phenotypic traits among different Gulin-niupicha populations

表4 2012年古蔺牛皮茶群体春、夏梢主要生化成分 Table 4 Biochemical composition of Gulin-niupicha in spring and summer of 2012

2.2.2 2013年春梢主要生化成分不同居群春梢的主要生化品质成分的含量也有较明显的差异( 表5)。茶多酚含量为19.04%~29.79%, 咖啡碱含量3.36%~4.28%, 游离氨基酸总量2.27%~4.96%, 儿茶素总量为104.73~175.01 mg g^-1, 酚氨比为3.84~ 12.38。茶氨酸是茶叶氨基酸组分中最重要且含量最高的氨基酸, 又是茶叶中的特征氨基酸, 其含量为10.98~24.13 mg g^-1, 多数居群间与对照也有一定差异, 居群D比E和CK分别高119.76%、80.21%。与2012年的测试结果相比, 茶多酚含量略降低, 咖啡碱含量呈现一定差异, 游离氨基酸含量均出现不同程度上升, 但居群D和C仍最高, 分别为4.96%和4.78%。表现出古蔺牛皮茶资源生化成分的多样性。

2.2.3 2013年春梢儿茶素茶叶中的儿茶素是茶叶中多酚类物质的主体成分, 也是具有保健功效的生理活性物质。儿茶素主要含8种组分物质, 从表6可知。8种儿茶素组分的含量各居群间与对照均有较明显的差异, 其中最具保健功效的酯型儿茶素(EGCG)的含量为45.75~82.80 mg g^-1, 居群B、F、A的含量均显著高于CK, 其余4个居群则显著低于CK。

表5 2013年古蔺牛皮茶群体春梢主要生化成分 Table 5 Biochemical composition of Gulin-niupicha in spring of 2013

表6 不同居群古蔺牛皮茶春梢儿茶素组分分析(2013年) Table 6 Analysis of catechin components about the tea plants in spring of 2013

2.3.2 古蔺牛皮茶居群遗传多态性和遗传结构

从表8可知, 古蔺牛皮茶7个居群的平均多态性为64.66%, Nei’s基因多态性( H)指标为0.254, Shannon’s遗传信息指数( I)为0.392。各居群内, Nei’s基因多态性( H)系数平均为0.203, Shannon’s遗传信息指数( I)平均为0.307。在7个居群中, 居群C的遗传多态性最高, H值为0.218, I值为0.333; 而居群B的遗传多态性最低, H值为0.186, I值为0.163。从遗传结构来看, 古蔺牛皮茶居群的基因遗传分化指数( G_st)为0.198, 表明有19.8%的遗传变异存在于7个居群间, 80.2%的变异存在于居群内; 也说明古蔺牛皮茶居群内遗传变异较高, 居群间的遗传变异较低。基于居群间遗传分化系数计算的每代迁移数(即基因流 N_m)为2.022。

表7 15对ISSR引物对76株古蔺牛皮茶扩增的多态性 Table 7 Polymorphism detected with 15 ISSR primers in 76 Gulin-niupicha individuals

表8 不同古蔺牛皮茶群体中的ISSR遗传多态性 Table 8 Genetic diversity of Gulin-niupicha distributed among different populations detected by ISSR

2.3.3 古蔺牛皮茶居群亲缘关系聚类分析利用UPGMA (unweighted pair-group method with arith-metic means)法, 得出聚类树状图( 图1)。在遗传相似系数为0.956时, 可以将7个居群分成3个组, 居群A、居群B和居群D组成第I组, 居群C和居群E组成第II组, 居群F和居群G组成第III组。7个居群体之间, 居群A和居群B的遗传相似系数最大, 为0.981; 居群E和居群G的遗传相似系数最小, 为0.879; 表明居群A和居群B之间亲缘关系最近, 而居群E和居群G亲缘关系最远。

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	图1 基于ISSR分子标记和Jaccard遗传相似性系数的亲缘关系UPGMA聚类图Fig. 1 UPGMA dendrogram of seven Gulin-niupicha populations based on ISSR

2.4 形态学特征与遗传距离的相关性分析

利用Mantel测试方法对形态学特征矩阵和ISSR分子标记数据矩阵作相关性分析。显示, 在形态学特征和遗传距离之间不存在相关性, 其Mantel相关系数 r=0.03。

表9 不同古蔺牛皮茶居群间的遗传相似系数 Table 9 Genetic similarity coefficient between populations of Gulin-niupicha

3 讨论

3.1 古蔺牛皮茶形态学特征与品质生化成分

古蔺牛皮茶在树型、树姿、成熟叶片和新梢形状上都表现出多样性, 很多地区茶树地方种质资源都表现出这一特征。王春梅等^{[ 14]}在研究崇州枇杷茶野生大茶树种质资源时, 发现居群内变异类型较多, 不同类型茶树间差异显著, 树体、芽叶和成熟新梢等生物学特征均表现出多样性; 蒋会兵等^{[ 15]}对云南10个地区的830份茶树种质资源分析发现, 云南茶树种质资源具有丰富的表型遗传多态性。茶树在漫长的历史过程中, 经历了古地质气候的变迁, 形成了自己独特的生长发育规律和遗传特性。了解这些特征、特性,可为种质资源的鉴定和评介提供依据, 同时在茶树栽培和育种中综合运用各项技术,以实现茶叶的优质、高产、高效^{[ 16]}。本研究对古蔺牛皮茶的形态学性状的初步探索, 积累了一些数据, 对该资源的鉴定、评价与利用具有一定的指导意义。但是, 需对其进行长期深入的研究, 才能充分掌握其特征特性和开发利用价值。

茶叶生化成分组成及其含量是茶叶色、香、味形成的物质基础, 也是茶叶品质鉴定的重要指标之一^{[ 17, 18]}。前人通过生化成分测定, 已鉴定出众多品质优异的茶树资源材料^{[ 19, 20, 21]}。古蔺牛皮茶群体中新梢品质生化成分呈现出特异和多样的特点, 7个供试居群中, B和A居群的茶多酚含量较高(春梢>27%, 夏梢>31%), 且儿茶素含量也高(春梢>150 mg g^-1, 夏梢>190 mg g^-1); 居群D和C的春茶游离氨基酸总量高于4.5%。酚氨比值是鉴定茶叶适制性的重要依据, 一般认为, 酚氨比值小于8适宜制绿茶; 酚氨比值在8~15之间, 红绿茶兼制; 大于15则适制红茶^{[ 22]}。7个居群的春茶比值值均在3.84~16.95之间, 表明古蔺牛皮茶可以用来选育优质绿茶、红茶品种或红绿兼制品种。然而,茶树的生长发育和代谢受诸多环境因素的影响, 内质生化成分也会随地理、气候、发育阶段不同而改变。因此, 对古蔺牛皮茶深入研究才能挖掘群体中有价值的种质资源, 而对现有古蔺牛皮茶资源进行有效的保护是深入研究和开发利用的前提。

3.2 古蔺牛皮茶居群的遗传多态性和遗传结构

本研究用15对ISSR引物对76份古蔺牛皮茶材料的遗传多态性进行分析,多态性位点百分比为90.4%, 高于季鹏章等^{[ 23]}利用ISSR研究云南古树茶10个居群的多态性位点百分比(56.5%~90.9%), 略低于宁静等^{[ 24]}研究的湘西“黄金茶”群体和李丹等^{[ 25]}研究的江华苦茶的多态性位点百分比(分别为91.22%和95.40%), 远低于刘本英等^{[ 26]}研究云南茶树资源的多态性(98.9%)。表明古蔺牛皮茶基因组DNA具有一般的多态性, 也反映出古蔺牛皮茶群体一定的遗传变异程度。76份古蔺牛皮茶材料的平均Nei’s基因多样性指数( H)为0.254, 平均Shannon’s信息指数( I)为0.392, 比姚明哲等^{[ 27]}研究我国茶树无性系品种多样性指数( H为0.23; I为0.38)略高, 这是由于我国选育茶树品种时, 同一地区的大部分育成品种都源于遗传背景比较相似的地方群体或亲缘关系比较近的亲本, 导致遗传基础相对单一; 同时, 良种的繁育主要采用无性繁殖技术, 也会导致无性系茶树遗传多样性较低。宁静等^{[ 24]}研究的湘西“黄金茶”地方群体的多态性指数( H为0.30, I为0.45)和李丹等^{[ 25]}研究的江华苦茶的多态性指数( H为0.38; I为0.56)都比古蔺牛皮茶居群高, 尤其是后者, 表明古蔺牛皮茶的在基因上多态性并不是很高。作为一个地方特有的茶树居群, 古蔺牛皮茶处于野生、半野生和栽培状态, 茶树的来源和生长发育已受到一定的人为影响, 比如茶农对具有优良农艺性状茶树会进行保护和培育, 而对农艺性状较差的茶树则较少关注, 这种人工选择会导致古蔺牛皮茶居群遗传背景变窄, 从而降低居群的遗传多态性^{[ 28]}, 这可能是古蔺牛皮茶对比湘西“黄金茶”群体和江华苦茶群体表现出较低基因多态性的原因之一。

植物居群的遗传结构是由一系列的作用形成的, 包括物种的长期进化(如分布的变化, 栖息地的分裂和种群隔离)、突变、繁殖体系以及基因流动和选择等^{[ 29]}。表型变异是基因型和环境共同作用的结果, 丰富的表型变异蕴藏着一定的遗传变异^{[ 30]}。本研究中古蔺牛皮茶居群间的遗传分化系数( G_st)是0.198, 这与杨阳等^{[ 31]}利用EST-SSR标记研究黄金茶群体遗传多样性及遗传分化分析结果相似。而对应的群体每代迁移数即基因流动估值( N_m)为2.022, 表现出中度的基因流动性。Wright^{[ 32]}认为, 如果居群间基因流指数 N_m>1, 基因流便能发挥均质化作用, 可有效防止不同地区亚居群间遗传分化的产生; 反之如果 N_m<1, 则表示各亚居群间基因流受阻, 居群走向遗传分化。本研究中古蔺牛皮茶各居群间的基因流指数大于1, 说明整个群体内遗传分化较低。不同区域居群之间的分化程度比较低, 也表明地理位置对其选择压力不大。

3.3 遗传距离与形态学特征的相关性

本研究中, 古蔺牛皮茶基于ISSR标记的遗传距离与形态学特征之间未表现出相关性。类似的结果在一些野生植物居群中也出现过, Lopes等^{[ 33]}在研究葡萄牙野生杨梅居群的遗传多态性时, 发现形态学性状和分子遗传特征之间不存在相关性( r=0.09), Tantasawat等^{[ 34]}在研究一年生豇豆时也发现类似的结果。造成这种结果的原因是多方面的, Pirkhezri等^{[ 35]}认为, 生物表现出来的外形特征可能是由多对等位基因共同决定的, 这些等位基因都会表现出相同或者相似的某一性状, 而这些等位基因和表型性状之间并不存在必然的相关性。此外, 本试验选择的ISSR引物本身存在一定局限性, 不可能将所有的决定表型性状的等位基因都扩增出来, 茶树生长的特殊环境也会对茶树生长发育产生影响, 从而影响茶树性状特征的表现。

4 结论

古蔺牛皮茶在形态学特征和品质生化成分上呈现出多态性, 但居群的平均Nei’s基因多样性指数( H)为0.254, 平均Shannon’s信息指数( I)为0.392, 表明遗传多态性较低。7个居群的UPGMA遗传相似性聚类显示, 在相似系数为0.956处可将之分为3个组。居群间的遗传分化系数( G_st)是0.198, 证明古蔺牛皮茶居群的遗传多样性主要分布在自然居群内。

The authors have declared that no competing interests exist.

作者已声明无竞争性利益关系。

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[28]	姜燕华, 段云裳, 王丽鸳, 周健, 曾建明, 成浩. 福建茶树品种的SSR分析及其人为选择的影响. 浙江林业科技, 2010, 30(3): 12-16 Jiang Y H, Duan Y S, Wang L Y, Zhou J, Zeng J M, Cheng H. SSR analysis on tea germplasm in Fujian Province and effect of variety selection. J Zhenjiang For Sci & Tech, 2010, 30(3): 12-16 (in Chinese with English abstract) [本文引用:1]
[29]	Schaal B A, Hayworth D A, Olsen K M, Rauscher J T, Smith W A. Phylogeo graphic studies in plants: problems and prospect. Mol Ecol, 1998, 7: 465-474 [本文引用:1] [JCR: 6.275]
[30]	吴会芳, 李作洲, 黄宏文. 湖北野生天麻的遗传分化及栽培天麻种质评价. 生物多样性, 2006, 14: 315-326 Wu H F, Li Z Z, Huang H W. Genetic differentiation among natural populations of Gastrodia elata (Orchidaceae) in Hubei and germplasm assessment of the cultivated populations. Biodiv Sci, 2006, 14: 315-326 (in Chinese with English abstract) [本文引用:1] [CJCR: 1.854]
[31]	杨阳, 刘振, 赵洋, 梁国强, 赵熙. 利用EST-SSR标记研究黄金茶群体遗传多样性及遗传分化. 茶叶科学, 2009, 29: 236-242 Yang Y, Liu Z, Zhao Y, Liang G Q, Zhao X. Genetic diversity and relationship of Huangjincha cultivar based on EST-SSR markers. J Tea Sci, 2009, 29: 236-242 (in Chinese with English abstract) [本文引用:1] [CJCR: 1.225]
[32]	Wright S. Evolution and the genetics of populations In Variability Within and Among Natural Populations. Chicago: University of PressUniversity of Chicago Press, 1978. pp149-156 [本文引用:1]
[33]	Lopes L, Olga S, Pereiar J A, Baptista P. Genetic diversity of Portuguese Arbutus unedo L. populations using leaf traits and molecular markers: an approach for conservation purposes. Sci Hort, 2012, 142: 57-67 [本文引用:1]
[34]	Tantasawat P, Trongchuen J, Prajongjai T, Seehalak W, Jittayasothorn Y. Variety identification and comparative analysis of genetic diversity in yardlong bean (Vigna unguiculata ssp. sesquipedalis) using morphological characters, SSR and ISSR analysis. Sci Hort, 2010, 124: 204-216 [本文引用:1]
[35]	Pirkhezri M, Hassani M E, Hadian J. Genetic diversity in different populations of Matricaria chamomilla L. growing in Southwest of Iran, based on morphological and RAPD markers. Res J Med Plant, 2010, 4: 1-13 [本文引用:1] [JCR: 0.879]

2005

0.0

... 它蕴藏着各种性状的遗传基因, 是选育新品种和发展农业生产的物质基础, 也是进行生物学研究的重要材料^[1] ...

... 3 形态学特征观察于2012年春季驻点观测, 根据NY/T 1312-2007农作物种质资源鉴定技术规程(茶树)^[4]的方法, 观测古蔺牛皮茶树体、芽叶及成熟叶片的性状及特征, 并按相应方法和要求^[1]记录于茶树种质资源数据采集表 ...

1981

0.0

0.717

... 我国西南地区的云贵川一带是茶树的发源地之一, 拥有丰富的野生、半野生和栽培的茶树种质资源^[2] ...

2013

0.0

... 目前有关古蔺牛皮茶的研究较少^[3], 未见对古蔺牛皮茶系统研究的报道 ...

2007

0.0

... 4 品质生化成分测定于2012年春、夏两季及2013年春季采摘茶树的标准一芽二叶制作生化样, 根据NY/T 1312-2007农作物种质资源鉴定技术规程(茶树)^[4]制定的方法, 测定古蔺牛皮茶的品质生化成分, 测定指标包括茶多酚^[7]、游离氨基酸^[8]和咖啡碱^[9] ...

2007

0.0

... 为方便后期的统计分析, 参照沈程文^[5]的方法量化处理形态学特征的质量指标(表2) ...

2002

0.0

2008

0.0

... 采用高效液相色谱法(HPLC)测定儿茶素和茶氨酸^[7] ...

2002

0.0

2002

0.0

2006

0.0

0.661

... 参照刘本英等^[10]的方法提取茶树基因组DNA ...

... 参照刘本英等^[10]的ISSR反应体系, 20 #cod#x003bc ...

2008

0.0

... 从加拿大哥伦比亚大学(University of British Columbia Biotechnology, UBS)报道的100条植物通用ISSR引物^[11]中挑选72条, 委托北京华大基因有限公司合成 ...

2005

0.0

1.854

罗晓莹, 唐光大, 许涵, 庄雪影, 郑文经, 郑明. 山茶科3种中国特有濒危植物的遗传多样性研究. 生物多样性, 2005, 13: 112-121

Luo

X Y

, Tang

G D

, Xu

, Zhuang

X Y

, Zheng

W J

, Zheng

Genetic diversity of three endemic and endangered species of the family Theaceae in Guangdong, China. Biodiv Sci, 2005, 13: 112-121 (in Chinese with English abstract)

Euryodendron excelsum, Apterosperma oblata, and Camellia changii are three species of the family Theaceae endemic to China. Genetic diversity of the three endangered species was studied using random amplified polymorphic DNA (RAPD). A total of 206－305 bands were generated by 20-24 random primers. Percentages of polymorphic loci(PPL) of the three species were 51.80％, 80.26％ and 38.83％, respectively, and coefficients of gene differentiation (G_ST) were 0.3566, 0.1713, and 0.1242, respectively. The results indicate that Apterosperma oblata, with wider natural distribution, has higher genetic diversity than the other two species with restricted natural distributions. Genetic variation mainly existed within populations; however, population genetic differentiation was obviously higher in Euryodendron excelsumthan in the other two species, which possibly resulted from habitat fragmentation. Based on the different genetic structure, we put forward some conservation strategies, including in situ preservation, establishing a conservation centre for endangered species of the family Theaceae in Guangdong, and developing fast propagation techniques.

1 College of Forestry, South China Agricultural University, Guangzhou 510642 2 Yangchun Forestry Research Institute, Yangchun, Guangdong 529600 3 Yangchun Forestry Bureau, Yangchun, Guangdong 529600

猪血木(Euryodendron excelsum)、圆籽荷(Apterosperma oblata)和杜鹃红山茶(Camellia changii)均为我国特有的山茶科珍稀濒危植物。本研究应用随机扩增多态性DNA(RAPD)检测方法，比较了这3个物种的遗传多样性。利用20－24个随机引物从猪血木、圆籽荷和杜鹃红山茶居群中分别获得206－305条RAPD谱带，多态位点百分率(PPL)分别为51.80％、80.26％和38.83％；居群间遗传分化指数G_ST分别为0.3566、0.1713、0.1242。结果显示：自然分布较广的圆籽荷的遗传多样性高于另外两种自然分布区狭窄的种类。3个物种的遗传变异均主要存在于居群内，但猪血木居群间的遗传分化明显高于圆籽荷和杜鹃红山茶，这可能与其生境的严重片断化有关。根据3个物种不同的遗传结构特点，建议加强现有自然居群的就地保护，促进居群自然更新；收集不同分布点的种源，尽快建立广东山茶科珍稀濒危植物的保育中心，深入开展人工快繁研究，扩大这些种群的数量。

... 参照罗晓莹等^[12]的方法, 利用POPGEN Version 1 ...

1965

0.0

... 居群每代迁移数N_m=(1-F_st)/4F_st^[13] ...

2012

0.0

0.525

... 王春梅等^[14]在研究崇州枇杷茶野生大茶树种质资源时, 发现居群内变异类型较多, 不同类型茶树间差异显著, 树体、芽叶和成熟新梢等生物学特征均表现出多样性 ...

2013

0.0

1.667

蒋会兵, 宋维希, 矣兵, 李友勇, 马玲, 陈林波, 田易萍, 段志芬, 刘本英, 梁名志. 云南茶树种质资源的表型遗传多样性. 作物学报, 2013, 39: 2000-2008

Jiang

H B

, Song

W X

, Yi

, Li

Y Y

, Ma

, Chen

L B

, Tian

Y P

, Duan

Z F

, Liu

B Y

, Liang

B Z.

Genetic diversity of tea germplasm resources in Yunnan province based on phenotypic characteristics. Acta Agron Sin, 2013, 39: 2000-2008 (in Chinese with English abstract)

We analyzed and compared the genetic diversity of main phenotypic characteristics in 830 tea germplasm, from ten different areas in Yunnan Province. The results showed Section Thea had the rich genetic diversity, the diversity index of descriptive traits was 0.64–1.40, with the average of 1.04, and relatively higher in young shoot color, the lowest in leaf base shape; the diversity index was 0.76–0.99, with the highest in Honghe prefecture, the second in Lincang and Wenshan prefectures, the lowest in Zhaotong prefecture. The variation coefficient of numerical traits was 10.27%–48.14%, with the average of 26.43%, and relatively higher in the ratio of polyphenol to amino acid, and the lowest in water extract; the variation coefficient was 16.09%–25.75%, with the highest in Wenshan and Honghe prefectures, and the lowest in Zhaotong and Dali. The genetic diversity and variation coefficient showed a tendency of Southeast>West>Northeastern in phenotypic characteristics. By cluster analysis, tea germplasm resources from ten regions was divided geographically into three groups, Zhaotong and Honghe for each group respectively, others for the third group. The results demonstrate an obviously ecogeographical differentiation among Southeast, West, and Northeast areas of tea germplasm in Yunnan, and Southeast is the distribution center with genetic diversity in phenotypic characteristics.

Tea Research Institute, Yunnan Academy of Agricultural Sciences / Engineering Research Centers of Tea Germplasm Resources Innovation and Its Matching Cultivated Techniques, Menghai 666201, China

对云南10个地区的830份茶树种质资源主要表型性状的遗传多样性分析和比较表明，云南茶树种质资源具有丰富的表型遗传多样性，13个描述型性状的遗传多样性指数在0.64~1.40之间，平均为1.04，以芽叶色泽较高，叶基较低；不同地区的茶树资源遗传多样性指数在0.76~0.99之间，以红河地区较高，其次是临沧和文山地区，以昭通地区较低。14个数值型性状的变异系数在10.27%~48.14%之间，平均为26.43%，以酚氨比较大，水浸出物含量较小；不同地区的茶树资源变异系数在16.09%~25.75%之间，以文山、红河地区较大，昭通、大理地区较小。茶树资源表型性状的遗传多样性和变异水平显示东南部>西部>东北部的趋势。通过聚类分析，10个地区的茶树资源划分为3个地理组，东北部的昭通和东南部的红河各单独聚为一组，其他8个地区聚为第3组，显示云南茶树资源存在东南部、西部、东北部生态地理的分化，初步推断东南部为云南茶树资源表型遗传多样性分布中心。

... 蒋会兵等^[15]对云南10个地区的830份茶树种质资源分析发现, 云南茶树种质资源具有丰富的表型遗传多态性 ...

2000

0.0

... 了解这些特征、特性,可为种质资源的鉴定和评介提供依据, 同时在茶树栽培和育种中综合运用各项技术,以实现茶叶的优质、高产、高效^[16] ...

2012

0.0

1.225

1964

0.0

1.225

1992

0.0

1.225

2001

0.0

1.225

2011

0.0

1.225

宋维希, 刘本英, 矣兵, 蒋会兵, 马玲, 段志芬, 孙雪梅, 汪云刚, 王平盛. 云南茶树优异种质资源的鉴定评价与筛选. 茶叶科学, 2011, 31: 45-52

Song

W X

, Liu

B Y

, Yi

, Jiang

H B

, Ma

, Duan

Z F

, Sun

X M

, Wang

Y G

, Wang

P S.

Identification, Evaluation and screening on elite and rare tea germplasms from Yunnan province. J Tea Sci, 2011, 31: 45-52 (in Chinese with English abstract)

从农艺性状、加工品质、生化成分、抗性等方面,对云南500份茶树种质资源进行了全面系统的鉴定和评价.采用近100项评定指标,筛选出52份优质资源,其中:四项常规成分含量均衡的32份;发芽特早生的2份;超常规水平的高多酚(>38%)含量5份、高咖啡碱(>5.2%)7份、低咖啡碱(<1%)的2份、高茶黄素(>1.6%)10份;水浸出物含量>48%的26份;抗根结线虫较强的2份;抗假眼小绿叶蝉较强的1份;抗咖啡小爪螨虫较强的2份;抗寒性较强的6份.

2008

0.0

... 大于15则适制红茶^[22] ...

2007

0.0

1.225

... 4%, 高于季鹏章等^[23]利用ISSR研究云南古树茶10个居群的多态性位点百分比(56 ...

2010

0.0

1.225

... 9%), 略低于宁静等^[24]研究的湘西#cod#x0201c ...

... 宁静等^[24]研究的湘西#cod#x0201c ...

2012

0.0

1.225

... 群体和李丹等^[25]研究的江华苦茶的多态性位点百分比(分别为91 ...

... 45)和李丹等^[25]研究的江华苦茶的多态性指数(H为0 ...

2010

0.0

1.667

刘本英, 李友勇, 唐一春, 王丽鸳, 成浩, 王平盛. 云南茶树资源遗传多样性与亲缘关系的ISSR分析. 作物学报, 2010, 36: 391-400

Liu

B Y

, Li

Y Y

, Tang

Y C

, Wang

L Y

, Cheng

, Wang

P S.

Genetic diversity and relationship of tea germplasm in Yunnan revealed by ISSR analysis. Acta Agron Sin, 2010, 36: 391-400 (in Chinese with English abstract)

Tea is an important beverage crop in the world. Yunnan region in China is the origin center of tea plants. Our knowledge on genetic diversity and relationship of tea germplasm in Yunnan province is critical to guide tea breeding. In order to provide theoretical information for using tea germplasm in tea breeding, we investigated the relationship and genetic diversity of the tea germplasm in Yunnan province. Eight species, including 134 tea varieties were used to detect the genetic variation by inter-simple sequence repeats (ISSR) analysis. The result showed that 475 DNA fragments among all 134 tea accessions were amplified, using 18 reliable ISSR primers, among which 470 DNA bands were polymorphic (PPB=98.9%). This indicated that a great amount of genetic polymorphism exists among tea germplasm tested. The genetic similarity (GS) among the tested genotypes ranged from 0.445 to 0.819, with an average of 0.512, indicating a wide gene pool among tea varieties in Yunnan. The cluster analysis presented that these resources were divided into three main groups using the unweighted pair-group method with arithmetic average (UPGMA) based on ISSR molecular marker data, but the dendrogram did not indicate clear division among tested varieties based on their geographical origin. Principal Component analysis (PCA) for ISSR data showed that PCA supported UPGMA clustering result, but showed more explicit relationships among the test accessions with different lays, orientationsand positions. The GS among 8 populations ranged from 0.850 to 0.987, with an average of 0.92, indicating that there existed a small variation of genetic diversity among different population. The findings of this research would be favorable for the further practice, such as tea breeding, the molecular genetic linkage mapping and the DNA fingerprint building of tea germplasm.

1 Research Center for Tea Germplasm and Improvement, Tea Research Institute of Chinese Academy of Agricultural Science, National Center for Tea Improvement, Hangzhou 310008, China; 2 Tea Research Institute of Yunnan Academy of Agricultural Science, Menghai 666201, China

以8个种群134份云南茶树资源为材料，应用ISSR标记方法，进行了遗传多样性与亲缘关系分析。结果表明，18个多态性ISSR引物对全部试验材料进行PCR扩增，共获得475条稳定的谱带，其中多态性谱带470条(占98.9%)，遗传多样性丰富；应用Nei-Li相似系数法估算了134个材料间的相似系数为0.445~0.819，平均为0.512，说明茶树资源间的遗传基础较宽；对134份茶树资源的分子系统聚类分析(UPGMA)将资源分为3大组，聚类结果与地理距离没有明显的相关性；主成分分析(PCA)表明主成分分析的结果与系统聚类基本一致，但是主成分分析更加直观清晰地显示各个材料间的亲缘关系；大厂茶等8个种群间的遗传相似系数于0.850~0.987间，平均为0.92，表明不同种群间的遗传差异较小。

... 40%), 远低于刘本英等^[26]研究云南茶树资源的多态性(98 ...

2007

0.0

1.667

姚明哲, 陈亮, 王新超, 赵丽萍, 杨亚军. 我国茶树无性系品种遗传多样性与亲缘关系的ISSR分析. 作物学报, 2007, 33: 598-604

Yao

M Z

, Chen

, Wang

X C

, Zhao

L P

, Yang

Y J.

Genetic diversity and relationship of clonal tea cultivars in China revealed by ISSR markers. Acta Agron Sin, 2007, 33: 598-604 (in Chinese with English abstract)

Tea is an important beverage crop in China. In recent years many clonal tea cultivars have been developed and widely replaced the diverse traditional tea population. Our knowledge on genetic diversity and relationship of these clonal tea cultivars is critical to guide tea breeding, but still limited. In this study, the genetic diversity and relationship of 36 clonal tea cultivars which mainly cultivated in China were analyzed with ISSR markers. Based on 20 ISSR primers, total 368 bands were generated, of which 367 (99.7%) were polymorphic. The average polymorphism information content (PIC) was 0.90, Nei’s gene diversity (H) and Shannon’s information index (I) were 0.23 and 0.38, respectively. Diversity of clonal cultivars within tea area was lower than that of in whole nation. The levels of diversity in Tea Area of Jiangnan and South China were higher than those of Southwest China. Analysis of molecular variance (AMOVA) showed that variance component among cultivars (94.4%) was far higher than that among regions (5.6%). The genetic similarity among all the tested clonal cultivars ranged from 0.58 to 0.84, averaging 0.69, which indicated that the genetic basis was relatively narrow. The cluster analysis based on ISSR markers revealed that cultivar “Jinxuan” from Chinese Taiwan was of the farthest distance with others from Chinese mainland in the dendrogram. Thirty-five clonal cultivars from Chinese mainland were clustered into a major group, and they could be further classified into three sub-groups except cultivar “Yihongzao”. The genetic relationship of all 36 tea cultivars revealed by the ISSR dendrogram would be of great help to accurately select the parents in tea breeding programs.

Laboratory for Tea Germplasm, Genetic Improvement and Molecular Biology / Tea Research Institute, Chinese Academy of Agricultural Sciences, Hangzhou 310008, Zhejiang, China

利用ISSR分子标记分析了我国36个主要茶树无性系品种的遗传多样性和亲缘关系。结果表明20个ISSR引物在供试品种中共扩增出368条谱带，其中多态性条带占总条带的99.7%，引物的多态性信息指数（PIC）平均达0.90。供试品种的基因多样性（H）和Shannon信息指数（I）分别为0.23和0.38。茶区内茶树品种的遗传多样性低于总体水平，江南和华南茶区主栽无性系品种的多样性高于西南茶区。AMOVA分析表明区域因素引起的变异（占5.6%）远小于品种因素（占94.4%）。供试品种间的相似系数介于0.58~0.84，平均为0.69，显示出我国茶树主栽品种的遗传基础已相对比较狭窄。ISSR聚类分析表明，中国台湾品种金萱与大陆品种的遗传距离较远，形成单独的个类。35个大陆品种聚成一个大类群，其中除宜红早形成独立的个类外，其他品种又聚为3个亚类群。亲缘关系树状图在分子水平上显示了我国主要茶树无性系品种间的亲缘关系，为今后茶树育种亲本的选配提供了理论依据。

... 392, 比姚明哲等^[27]研究我国茶树无性系品种多样性指数(H为0 ...

2010

0.0

姜燕华, 段云裳, 王丽鸳, 周健, 曾建明, 成浩. 福建茶树品种的SSR分析及其人为选择的影响. 浙江林业科技, 2010, 30(3): 12-16

Jiang

Y H

, Duan

Y S

, Wang

L Y

, Zhou

, Zeng

J M

, Cheng

SSR analysis on tea germplasm in Fujian Province and effect of variety selection. J Zhenjiang For Sci & Tech, 2010, 30(3): 12-16 (in Chinese with English abstract)

采用36对引物对60份福建省茶树品种进行了SSR分析,对所获数据的聚类分析和遗传结构分析都将参试材料分为两大类群,一大类是基本未获推广应用的未审(认)定地方品种,另一大类则包含品质性状优异而获得较大面积推广应用的审(认)定地方品种和现代育成品种;他们的多态性从高到低依次为未审(认)定地方品种＞育成品种＞审(认)定地方品种,表明带有相同或相似目的的人为选择,导致降低了推广使用的茶树品种的遗传多样性.

... 作为一个地方特有的茶树居群, 古蔺牛皮茶处于野生、半野生和栽培状态, 茶树的来源和生长发育已受到一定的人为影响, 比如茶农对具有优良农艺性状茶树会进行保护和培育, 而对农艺性状较差的茶树则较少关注, 这种人工选择会导致古蔺牛皮茶居群遗传背景变窄, 从而降低居群的遗传多态性^[28], 这可能是古蔺牛皮茶对比湘西#cod#x0201c ...

1998

6.275

0.0

... 植物居群的遗传结构是由一系列的作用形成的, 包括物种的长期进化(如分布的变化, 栖息地的分裂和种群隔离)、突变、繁殖体系以及基因流动和选择等^[29] ...

2006

0.0

1.854

吴会芳, 李作洲, 黄宏文. 湖北野生天麻的遗传分化及栽培天麻种质评价. 生物多样性, 2006, 14: 315-326

H F

, Li

Z Z

, Huang

H W.

Genetic differentiation among natural populations of Gastrodia elata (Orchidaceae) in Hubei and germplasm assessment of the cultivated populations. Biodiv Sci, 2006, 14: 315-326 (in Chinese with English abstract)

Gastrodia elata (Orchidaceae) is a rare and endangered medicinal plant. Seven ISSR primers were used to examine population genetic parameters of 483 samples across eight natural populations and six cultivated populations of G. elata. The seven primers amplified a total of 77 unambiguous and reproducible bands, among which 64 bands were polymorphic and the percentage of polymorphic loci was PPB=83.12%. The results showed that genetic variation in the natural populations was significantly higher than that in the cultivated populations (PPB=59.09% vs. PPB=35.71%, Ae=1.29 vs. Ae=1.16, H=0.176 vs. H=0.100 and Shannon’s I=0.270 vs. I=0.155), suggesting that the cultivated populations are narrowly genetic-based and genetically uniform. UPGMA cluster analysis showed that natural populations and cultivated populations were distinctly separated into two groups. Coefficient of genetic differentiation among eight natural popula-tions was 0.2558 (G_ST) which was well in accordance with the result by AMOVA analysis (27.25% of the to-tal genetic variation resided among populations), indicating that natural populations were genetically differ-entiated to a certain extent. Gene flow among natural populations was relatively weak (N_m=1.4547), which might have caused the genetic differentiation among the populations. It is interesting to note that natural populations tended to cluster by geographical locations as revealed in UPGMA dendrogram, while the Mantel test showed no significant correlation between genetic and geographic distances among natural popula-tions(r=0.1669, P=0.2110). This incongruence suggested that the current genetic differentiation among natu-ral populations might have resulted from combined effects, including those of life history traits, geographical isolation and human disruption. The genetic uniformity found in cultivated populations might reflect the loss of genetic diversity caused by a genetic bottleneck during human domestication and clonal reproduction over generations. The narrow genetic base as revealed in cultivated G. elata and the genetic differentiation be-tween cultivated and natural populations might have resulted from a cutoff of gene exchange when domesti-cation began.

Wuhan Botanical Garden, Chinese Academy of Sciences, Wuhan 430074

采用7条ISSR引物对天麻(Gastrodia elata) 8个自然居群和6个人工栽培居群共483个样本的居群遗传多样性进行了初步检测, 共检测出清晰、重复性好的DNA带77条, 其中64条为多态性带, 总多态位点百分比PPB=83.12%。遗传多样性分析结果表明: 天麻自然居群的遗传多样性参数分别为：多态位点百分比PPB=59.09%，有效等位基因数Ae =1.29，Nei’s遗传多样度H=0.176，Shannon’s多态信息指数I=0.270，明显高于人工栽培居群(PPB= 35.71%，Ae=1.16，H=0.100，I=0.155), 揭示出栽培居群存在明显的遗传基础狭窄和遗传均质性问题。UPGMA聚类分析表明, 自然居群与栽培居群存在明显的分化而分别聚为两大类群。自然居群间基因分化系数G_ST=0.2558, 与AMOVA分析所揭示的居群间遗传变异量占总变异的27.25％的结果相近, 说明天麻自然居群间亦存在一定程度的遗传分化; 居群间基因流(N_m)为1.4547, 相对较弱, 可能对自然居群的遗传分化有一定影响。自然居群聚类结果显示出一定程度的地理区域聚类趋势, 但Mantel检验表明自然居群间遗传距离与地理距离并不存在显著相关(r=0.1669, P=0.2110), 揭示出天麻自然居群的分化现状可能是其生活史特性、地理隔离与人为破坏综合作用的结果。栽培居群的遗传均质化趋势, 揭示了引种驯化的瓶颈效应和长期无性繁育所导致的遗传多样性丧失, 也反映出栽培天麻种质的遗传基础狭窄。而栽培居群与自然居群间存在着明显的遗传分化, 反映天麻栽培居群与自然居群间可能存在基因流的阻断。

... 表型变异是基因型和环境共同作用的结果, 丰富的表型变异蕴藏着一定的遗传变异^[30] ...

2009

0.0

1.225

杨阳, 刘振, 赵洋, 梁国强, 赵熙. 利用EST-SSR标记研究黄金茶群体遗传多样性及遗传分化. 茶叶科学, 2009, 29: 236-242

Yang

, Liu

, Zhao

, Liang

G Q

, Zhao

Genetic diversity and relationship of Huangjincha cultivar based on EST-SSR markers. J Tea Sci, 2009, 29: 236-242 (in Chinese with English abstract)

利用19对茶树EST-SSR引物对黄金茶群体的38个单株进行了遗传多样性和亲缘关系分析.19对引物共检测到等位位点37个,每个引物1～3个,平均1.95个;期望杂合度(He)在0.03～0.65之间,平均值0.32;观测杂合度(Ho)在0～0.97之间,平均值0.33;群体内的Shannon指数0.55.以上结果均说明黄金茶群体的遗传多样性较低.基于EST-SSR数据以SAHN邻接法对供试种质资源进行UPGMA遗传相似性聚类,并绘制树状聚类图,按相似系数0.77可将参试的38份种质资源分为六大类.根据不同单株间的相似系数可为黄金茶群体的遗传改良提供一定参考.同时,AMOVA分析表明,黄金茶群体的遗传变异21.77%发生在种群间,78.23%发生于单株间,不同区域的种群间基因流为1.80,这可以为黄金茶群体的保护提供一定理论依据.

... 198, 这与杨阳等^[31]利用EST-SSR标记研究黄金茶群体遗传多样性及遗传分化分析结果相似 ...

1978

0.0

... Wright ^[32]认为, 如果居群间基因流指数N_m#cod#x0003E ...

2012

0.0

... 类似的结果在一些野生植物居群中也出现过, Lopes等^[33]在研究葡萄牙野生杨梅居群的遗传多态性时, 发现形态学性状和分子遗传特征之间不存在相关性(r=0 ...

2010

0.0

... 09), Tantasawat等^[34]在研究一年生豇豆时也发现类似的结果 ...

2010

0.879

0.0

... 造成这种结果的原因是多方面的, Pirkhezri等^[35]认为, 生物表现出来的外形特征可能是由多对等位基因共同决定的, 这些等位基因都会表现出相同或者相似的某一性状, 而这些等位基因和表型性状之间并不存在必然的相关性 ...