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肾茶种质资源遗传多样性的ISSR分析



全 文 :收稿日期:2015 - 02 - 01
基金项目:海南省星火产业带专项“海南肾茶高效栽培与示范”(HNXH201319) ,海南省产学研一体化专项“海
南木棉产业发展关键技术集成与示范”(CXY20140027 ) ,国家级大学生创新创业训练计划项目“爱
吾园生物除醛创艺坊”(201421) ,海南省中西部高校提升综合实力工作资金项目“农艺应用型本科
实践教学创新团队”(201417) ,海南大学应用科技学院(儋州校区)基金项目“裂掌榕种质资源调查
及资源圃的建立”(HYK1524)
作者简介:罗灿(1988 -) ,男,硕士研究生;研究方向:植物资源与利用. E-mail:wadmrlc@ 126. com
通信作者:于旭东(1973 -) ,男,副教授,博士研究生,研究方向:资源植物学. E-mail:doeast@ 163. com
第 6 卷 第 2 期 热 带 生 物 学 报 Vol. 6 No. 2
2015 年 6 月 JOURNAL OF TROPICAL BIOLOGY Jun. 2015
文章编号:1674 - 7054(2015)02 - 0204 - 019
肾茶种质资源遗传多样性的 ISSR分析
罗 灿1,于旭东1,2,3,吴繁花2,高 鑫3,王童欣2
(1 海南大学 园艺园林学院 /热带作物种质资源保护与开发利用教育部重点实验室,海南 海口 570228 ;
2 海南大学 农学院 /热带珍稀植物资源开发与利用实验室,海南 海口 570228;
3 海南大学 应用科技学院,海南 儋州 571737)
摘 要:为了探讨中国肾茶种质资源的遗传多样性,本研究采用 100 条 ISSR引物对 87 份肾茶种质资源进行
试验分析,发现 20 条引物能扩增出清晰条带。在 87 份供试材料中,20 个 ISSR引物共产生 144 条扩增条带,
其中多态性条带 120 条,多态率 83. 3%,平均每个 ISSR标记能产生 6. 0 条多态性片段。87 份材料间的遗传
相似系数变化范围为 0. 48 ~ 0. 98,但 6 份栽培品种间遗传相似系数为 0. 88 ~ 0. 98,显示出栽培种的遗传基础
相对狭窄。利用 UPGMA聚类分析,供试材料可聚为 2 类,其中栽培品种单独聚成 1 类,野生材料聚为 1 类。
结果显示,中国野生肾茶种质资源遗传多样性丰富,具有较高的开发利用价值。
关键词:肾茶;CTAB法;ISSR;遗传多样性
中图分类号:S 326 文献标志码:A
肾茶[Clerodendranthus spicatus (Thunb.)C. Y. W u]为唇形科肾茶属植物,别名猫须草、猫须公,云
南傣族称肾茶为“雅糯妙”。文献中的 Orthosiphon aristatus 或 Orthosiphon stamineus,也均为肾茶[1]。肾茶
属有 5 种,主要分布在热带地区,原产于印度尼西亚、印度、缅甸、菲律宾、澳大利亚等[2],我国福建、台湾、
广东、广西、云南西双版纳、香港、四川米易等地均有引种栽培[3 - 4]。肾茶全草入药,全年采收,味甘、淡、微
苦、性凉,具有清热利湿,通淋排石的功效,有 2 000 多年的使用历史[5 - 7]。民间认为其全草具有利尿、抗
菌、消炎、溶石、排石和抗肿瘤作用[8 - 9],尤其对肾结石、尿路结石、胆结石、胆囊炎、风湿性关节炎及咽炎
有特殊疗效[10]。其主要化学成分有:黄酮类、酚性化合物、二萜类、三萜类、木脂素类、色原烯类、烷基糖
苷类、甾体皂苷类、蒽醌类、氨基酸、多肽、蛋白质及维生素、矿物质和有机酸等[11 - 14]。简单重复序列区间
(inter-simple sequence repeats,ISSR)PCR技术是 Zietkiewicz 等[15]提出的一种 DNA 分子标记,由于其在
SSR的 3端或 5端锚定 1 ~ 4 个简并碱基,所用的引物序列比 RAPD 的引物序列长,退火温度较高[16 - 18],
该技术不但具有 RAPD 技术的优点,又可克服 SSR 和 RAPD 技术的某些缺点,已应用于黄麻[19]、小
麦[20]、银杏[21]等植物遗传多样性与亲缘关系的研究。王英等 [22]采用 ISSR 分子标记技术对 96 份甘蔗种
质资源的遗传多样性进行了分析,结果表明其种质的遗传相似系数在 0. 15 ~ 0. 19 之间,证明 ISSR标记能
够很好揭示种质的遗传多样性。Behera 等[23]用 15 个 ISSR 引物对 38 份印度苦瓜(Momordica charantia
var. charantia,and var. muricata)进行遗传多样性分析,结果表明,苦瓜栽培品种与野生材料的起源不同,
这为苦瓜种内遗传分析和育种提供了依据。目前,肾茶在我国的种类尚不确定,海南植物志[24]记载肾茶
中国约有 2 种,海南有 1 种。我国肾茶分子方面的研究尚未见报道,笔者通过对收集的 87 份肾茶种质资
源进行 ISSR分子标记,以期揭示其遗传特性,为肾茶的种类鉴定、品种选育、产品的开发和应用提供理论
依据。
1 材料与方法
1. 1 材料
1. 1. 1 供试材料 实验材料来自海南大学儋州校区农学院教学科研基地迁地保育的肾茶种质资源圃。
将收集的 87 份肾茶(表 1)资源进行繁育,2 个月后长出新的嫩叶作为供试材料,选取健康无病斑的嫩叶
若干,装入编号袋,- 80 ℃条件下保存备用。
表 1 供试材料的来源
Tab. 1 Sources of C. spicatus accessions used in this study
序号 采集号 采集时间 采集地 经度 纬度 海拔 /m
S1 BW1 - 1 2013 - 08 - 05 海南霸王岭 109°633. 25″ 19°634. 56″ 225
S2 BW1 - 2 2013 - 08 - 05 海南霸王岭 109°633. 35″ 19°634. 97″ 245
S3 BW1 - 3 2013 - 08 - 05 海南霸王岭 109°633. 48″ 19°624. 58″ 263
S4 BW1 - 4 2013 - 08 - 05 海南霸王岭 109°634. 28″ 19°638. 29″ 274
S5 BW1 - 5 2013 - 08 - 05 海南霸王岭 109°631. 29″ 19°634. 60″ 308
S6 BW1 - 6 2013 - 08 - 05 海南霸王岭 109°633. 20″ 19°636. 41″ 313
S7 BW1 - 7 2013 - 08 - 05 海南霸王岭 109°638. 31″ 19°637. 62″ 323
S8 BW1 - 8 2013 - 08 - 05 海南霸王岭 109°633. 32″ 19°667. 43″ 334
S9 BW1 - 9 2013 - 08 - 05 海南霸王岭 109°653. 83″ 19°687. 22″ 347
S10 BW1 - 10 2013 - 08 - 05 海南霸王岭 109°633. 94″ 19°734. 15″ 366
S11 BW1 - 11 2013 - 08 - 05 海南霸王岭 109°635. 35″ 19°734. 36″ 388
S12 BW1 - 12 2013 - 08 - 05 海南霸王岭 109°633. 26″ 19°737. 14″ 405
S13 BW1 - 13 2013 - 08 - 05 海南霸王岭 109°643. 17″ 19°744. 25″ 422
S14 BW1 - 14 2013 - 08 - 05 海南霸王岭 109°643. 98″ 19°754. 33″ 456
S15 BW2 - 1 2013 - 08 - 06 海南霸王岭 109°652. 95″ 19°541. 12″ 269
S16 BW2 - 2 2013 - 08 - 06 海南霸王岭 109°655. 18″ 19°557. 20″ 282
S17 BW2 - 3 2013 - 08 - 06 海南霸王岭 109°653. 87″ 19°565. 20″ 305
S18 BW2 - 4 2013 - 08 - 06 海南霸王岭 109°652. 45″ 19°589. 31″ 316
S19 BW2 - 5 2013 - 08 - 06 海南霸王岭 109°652. 99″ 19°512. 25″ 333
S20 BW3 - 1 2013 - 08 - 08 海南霸王岭 109°1133. 12″ 19°216. 09″ 528
S21 BW3 - 2 2013 - 08 - 08 海南霸王岭 109°1133. 64″ 19°223. 10″ 497
S22 BW3 - 3 2013 - 08 - 08 海南霸王岭 109°1133. 33″ 19°212. 24″ 532
S23 BW3 - 4 2013 - 08 - 08 海南霸王岭 109°1133. 15″ 19°215. 12″ 488
S24 BW3 - 5 2013 - 08 - 08 海南霸王岭 109°1134. 10″ 19°219. 54″ 557
S25 BW3 - 6 2013 - 08 - 08 海南霸王岭 109°1134. 17″ 19°222. 14″ 540
502第 2 期 罗 灿等:肾茶种质资源遗传多样性的 ISSR分析
续表 1 Continued tab. 1
S26 BW3 - 7 2013 - 08 - 08 海南霸王岭 109°1133. 88″ 19°238. 67″ 511
S27 BW3 - 8 2013 - 08 - 08 海南霸王岭 109°1134. 67″ 19°215. 95″ 489
S28 BW4 - 1 2013 - 08 - 10 海南霸王岭 109°1130. 11″ 19°21. 92″ 427
S29 BW4 - 2 2013 - 08 - 10 海南霸王岭 109°1130. 15″ 19°22. 12″ 398
S30 BW4 - 3 2013 - 08 - 10 海南霸王岭 109°1130. 56″ 19°21. 45″ 388
S31 BW4 - 4 2013 - 08 - 10 海南霸王岭 109°1130. 84″ 19°21. 35″ 443
S32 BW4 - 5 2013 - 08 - 10 海南霸王岭 109°1130. 32″ 19°21. 46″ 450
S33 BW4 - 6 2013 - 08 - 10 海南霸王岭 109°1130. 65″ 19°21. 66″ 427
S34 BW4 - 7 2013 - 08 - 10 海南霸王岭 109°1131. 17″ 19°22. 38″ 422
S35 BW4 - 8 2013 - 08 - 10 海南霸王岭 109°1130. 74″ 19°22. 78″ 435
S36 BW4 - 9 2013 - 08 - 10 海南霸王岭 109°1130. 43″ 19°22. 34″ 439
S37 BW4 - 10 2013 - 08 - 10 海南霸王岭 109°1131. 55″ 19°22. 99″ 427
S38 BW5 - 1 2013 - 08 - 12 海南霸王岭 109°1129. 18″ 19°24. 08″ 444
S39 BW5 - 2 2013 - 08 - 12 海南霸王岭 109°1129. 15″ 19°24. 12″ 447
S40 BW5 - 3 2013 - 08 - 12 海南霸王岭 109°1129. 25″ 19°24. 15″ 436
S41 BW5 - 4 2013 - 08 - 12 海南霸王岭 109°1129. 22″ 19°24. 18″ 434
S42 BW5 - 5 2013 - 08 - 12 海南霸王岭 109°1129. 35″ 19°24. 24″ 430
S43 BW5 - 6 2013 - 08 - 12 海南霸王岭 109°1129. 63″ 19°24. 44″ 426
S44 BW5 - 7 2013 - 08 - 12 海南霸王岭 109°1129. 54″ 19°24. 88″ 422
S45 BW5 - 8 2013 - 08 - 12 海南霸王岭 109°1129. 14″ 19°25. 15″ 418
S46 BW5 - 9 2013 - 08 - 12 海南霸王岭 109°1129. 26″ 19°25. 36″ 414
S47 BW5 - 10 2013 - 08 - 12 海南霸王岭 109°1130. 27″ 19°25. 39″ 410
S48 BW5 - 11 2013 - 08 - 12 海南霸王岭 109°1129. 18″ 19°25. 48″ 406
S49 BW5 - 12 2013 - 08 - 12 海南霸王岭 109°1128. 15″ 19°25. 07″ 402
S50 BW5 - 13 2013 - 08 - 12 海南霸王岭 109°1128. 34″ 19°25. 20″ 398
S51 BW5 - 14 2013 - 08 - 12 海南霸王岭 109°1128. 47″ 19°25. 28″ 394
S52 YG1 - 1 2013 - 09 - 13 海南鹦哥岭 109°1847. 90″ 18°5959. 50″ 445
S53 YG1 - 2 2013 - 09 - 13 海南鹦哥岭 109°1847. 44″ 18°5959. 65″ 432
S54 YG1 - 3 2013 - 09 - 13 海南鹦哥岭 109°1847. 67″ 18°5959. 62″ 423
S55 YG1 - 4 2013 - 09 - 13 海南鹦哥岭 109°1847. 91″ 18°5959. 64″ 467
S56 YG1 - 5 2013 - 09 - 13 海南鹦哥岭 109°1847. 97″ 18°5959. 76″ 456
S57 YG1 - 6 2013 - 09 - 13 海南鹦哥岭 109°1847. 87″ 18°5959. 71″ 482
S58 YG1 - 7 2013 - 09 - 13 海南鹦哥岭 109°1847. 53″ 18°5959. 86″ 463
S59 YG1 - 8 2013 - 09 - 13 海南鹦哥岭 109°1847. 58″ 18°5959. 97″ 465
S60 YG2 - 1 2013 - 09 - 15 海南鹦哥岭 109°1848. 91″ 18°5959. 13″ 453
S61 YG2 - 2 2013 - 09 - 15 海南鹦哥岭 109°1848. 34″ 18°5959. 16″ 451
S62 YG2 - 3 2013 - 09 - 15 海南鹦哥岭 109°1848. 65″ 18°5959. 32″ 446
S63 YG2 - 4 2013 - 09 - 15 海南鹦哥岭 109°1848. 55″ 18°5959. 10″ 441
S64 YG2 - 5 2013 - 09 - 15 海南鹦哥岭 109°1848. 85″ 18°5959. 90″ 436
602 热 带 生 物 学 报 2015 年
续表 1 Continued tab. 1
S65 YG2 - 6 2013 - 09 - 15 海南鹦哥岭 109°1848. 34″ 18°5959. 87″ 459
S66 YG2 - 7 2013 - 09 - 15 海南鹦哥岭 109°1848. 39″ 18°5959. 67″ 444
S67 HNRDZWY1 -1 2013 - 07 - 22 海南热带植物园 109°3010. 42″ 19°3044. 02″ 137
S68 HNRDZWY1 -2 2013 - 07 - 22 海南热带植物园 109°3010. 43″ 19°3044. 03″ 137
S69 GXYYZWY1 -1 2013 - 11 - 21 广西药用植物园 108°2211. 23″ 22°5129. 32″ 87
S70 GXYYZWY1 -2 2013 -11 -22 广西药用植物园 108°2211. 24″ 22°5129. 33″ 87
S71 XSBN1 -1 2013 -10 -09 云南西双版纳植物园 101°1927. 71″ 21°5217. 23″ 864
S72 XSBN1 -2 2013 -10 -09 云南西双版纳植物园 101°1927. 71″ 21°5217. 24″ 864
S73 YG3 -1 2013 -09 -20 海南鹦哥岭 109°1860. 31″ 18°5958. 57″ 456
S74 YG3 -2 2013 -09 -20 海南鹦哥岭 109°1860. 23″ 18°5958. 48″ 445
S75 YG3 -3 2013 -09 -20 海南鹦哥岭 109°1860. 67″ 18°5958. 55″ 467
S76 YG3 -4 2013 -09 -20 海南鹦哥岭 109°1860. 89″ 18°5958. 53″ 453
S77 YG3 -5 2013 -09 -20 海南鹦哥岭 109°1860. 13″ 18°5958. 22″ 459
S78 YG4 -1 2013 -09 -22 海南鹦哥岭 109°1860. 90″ 18°5958. 44″ 460
S79 YG4 -2 2013 -09 -22 海南鹦哥岭 109°1860. 75″ 18°5958. 80″ 461
S80 YG4 -3 2013 -09 -22 海南鹦哥岭 109°1859. 46″ 18°5958. 90″ 462
S81 YG4 -4 2013 -09 -22 海南鹦哥岭 109°1859. 97″ 18°5958. 14″ 464
S82 YG4 -5 2013 -09 -22 海南鹦哥岭 109°1859. 91″ 18°5959. 56″ 487
S83 YG4 -6 2013 -09 -22 海南鹦哥岭 109°1859. 89″ 18°5959. 07″ 449
S84 YG4 -7 2013 -09 -22 海南鹦哥岭 109°1859. 80″ 18°5959. 78″ 435
S85 YG4 -8 2013 -09 -22 海南鹦哥岭 109°1859. 85″ 18°5959. 79″ 478
S86 WZ1 -1 2013 -08 -17 海南五指山 109°3141. 08″ 18°482. 58″ 561
S87 WZ1 -2 2013 -08 -17 海南五指山 109°3141. 23″ 18°482. 30″ 553
1. 1. 2 试剂与引物 氯化钠、醋酸钠、β-巯基乙醇、Tris-平衡酚、无水乙醇、异戊醇和氯仿等均为国产分析纯;
CTAB,EDTA,Mark2 000,Mark15 000,RNA 酶和琼脂糖均购于北京 Trangen company;10 × buffer,MgCl2(25
mmol·L -1) ,dNTP(10 mmol·L -1) ,Taq 酶(2. 5 U·uL -1)无菌水均购于北京 TianGen;ISSR引物由上海英
潍捷基合成。提取液配制根据 Chaudhry 等[25]的方法进行改良,包括 0. 45 mol·L -1葡萄糖,0. 1 mol·L -1
Tris-HCl (pH 8. 0) ,5 mmol·L -1 EDTA (pH 8. 0) ,2%(W/V)聚乙烯吡咯烷酮(PVP-36) ,2%(V /V)巯基乙
醇。裂解液的配制根据 Vrohbi等[26]的方法改良而来,包括 2%(W/V)CTAB,2%(W/V)PVP-36,1. 4 mol·
L -1 NaCl,20 mmol·L -1,EDTA (pH 8. 0) ,0. 1 mol·L -1 Tris-HCl (pH 8. 0) ,2%(V /V)巯基乙醇。TE 缓冲
液为:10 mmol·L -1 Tris-HCl,1 mmol·L -1 EDTA (pH 8. 0)。V酚 ∶ V氯仿 ∶ V异戊醇 =25∶ 24∶ 1,V氯仿 ∶ V异戊醇 =
24∶ 1,乙醇 70%,100%,醋酸钠(pH 5. 2)为 3 mol·L -1,异丙醇为 100%。
1. 1. 3 主要仪器 德国 EPPENDROF冷冻离心机、德国 EPPENDROF Biophotometer plus核算蛋白测定仪、德
国 TGRADIENT梯度 PCR仪、德国 BiotraPCR仪、北京百亿新凝胶成像系统(Tanon4100)等。
1.2 方法
1. 2. 1 基因组 DNA的提取 将 CTAB法改良用于提取肾茶的 DNA。(1)选取 4 ~ 6 片嫩叶,放入预冷的研
钵中,用液氮研磨至粉状,将粉末迅速装入灭菌的 2 mL离心管中,加入细胞提取液(改良)1 000 μL(4 ℃保
存)和 20 μL的 β-巯基乙醇混匀,在冰上放置 15 min,4 ℃,4 500 r·min -1离心 5 min;(2)弃上清,加入预热
65 ℃的 CTAB裂解液(改良)800 μL和 20 μL的 β-巯基乙醇混匀,将离心管放置于 65 ℃水浴锅中,每 5 min
上下翻转 30次,40 min后取出,冷却至常温;(3)加入等体积酚 -氯仿 -异戊醇混合液,摇晃至浑浊,在常温
702第 2 期 罗 灿等:肾茶种质资源遗传多样性的 ISSR分析
下放置 5 min,15 ℃,12 000 r·min -1离心 15 min;(4)取上清(约 900 μL) ,加入等体积的氯仿 -异戊醇混合
液,轻摇混匀,在冰上放置 5 min,4 ℃,12 000 r·min -1离心10 min;(5)重复步骤(4);(6)取上清(约600 μL),加
入 2 μL RNA酶,在 37 ℃水浴 30 min;(7)加入 1 /10体积 NaAC,加入 2倍体积无水乙醇混匀,在 -20 ℃静置 60
min,4 ℃,9 000 r·min -1离心 10 min;(8)弃上清,用 75%的乙醇洗涤沉淀,常温 10 000 r·min -1离心 5 min:(9)
重复步骤(8);(10)弃上清,置于超净工作台里对 DNA进行风干 20 ~30 min; (11)加入 50 μL TE缓冲液溶解沉
淀,4 ℃保存待测。
1. 2. 2 ISSR引物筛选 ISSR 引物为加拿大哥伦比亚大学公布的 100 条引物序列,由英潍捷基(上海)
贸易有限公司合成。选用 S2(海南霸王岭)、S59(海南鹦哥岭)、S72(云南西双版纳)3 份材料作为 DNA
模板,并从中筛选出 20 条扩增条带清晰,多态性较好的引物。用这 3 份材料 DNA模板对引物进行退火温
度的筛选,退火温度设定为 49 ~ 59 ℃,PCR仪自动生成 12 个退火温度,分别为 49. 0,49. 2,49. 0,51. 0,52.
3,53. 4,54. 6,55. 8,57. 0,58. 1,58. 8,59. 0 ℃,PCR产物用 1. 0%的琼脂糖凝胶电泳检测。
1. 2. 3 ISSR扩增反应体系的确立 ISSR扩增反应体系经过优化①后确定为 20 μL,反应体系如下:10 ×
buffer 2. 0 μL,MgCl2 2. 0 μL,dNTPs 1. 4 μL,ISSR引物(10 μmol·L
-1)1. 0 μL,Taq 酶(2. 5 U·μL -1)1. 0
μL,DNA模板(50 ng·μL -1)2. 0 μL,无菌水 10. 6 μL。PCR 反应程序为 94 ℃预变性 7 min;94 ℃ 变性 30
s;49 ~ 59 ℃(依据 1. 2. 2 结果确定)退火 30 s,72 ℃延伸 90 s,35 个循环;72 ℃延伸 10 min;4 ℃保存。
PCR产物在含 TBE的琼脂糖凝胶中电泳 45 min,电压 120 V。以 Marker2000 相对分子质量为标准对照,
用凝胶成像系统观察拍照记录,见图 1。
1. 3 数据处理 根据电泳结果记录清晰可重复的 ISSR 条带。对于同一条引物的扩增产物,迁移率相同
的条带记为 1 个位点,扩增阳性赋值为 1,阴性赋值为 0,根据统计条带的结果建立原始数据(0,1)矩阵。
统计分析采用 NTSYS2. 1 软件系统进行,利用 UPGMA聚类分析。
802 热 带 生 物 学 报 2015 年
① 罗灿. 87 份肾茶属植物种质资源亲缘关系研究[D],海南大学,2015.
2 结果与分析
2. 1 基因组 DNA的遗传多态性 从 UBC公布的 100 条引物中筛选出 20 条能扩增出条带清晰、多态性
及重复效果较好的 ISSR引物(表 2) ,通过对肾茶基因组 DNA 进行 PCR 扩增,PCR 扩增条带相对分子质
量一般在 100 ~ 2 000 bp(图 1)。用筛选出的 20 个 ISSR引物共扩增出 144 条谱带,平均每个引物扩增出
7. 2 条谱带,其中存在多态性的谱带有 120 条,多态性条带的比例(PPB)达 83. 33%(表 2)。结果表明肾
茶种质资源的遗传多样性丰富,ISSR标记可以较好地揭示出肾茶种质资源种内的遗传差异和亲缘关系。
表 2 引物序列及其扩增结果
Tab. 2 List of primers and their sequences as well as the amplification results
引物
Primer
引物序列 5'- 3'
Primer sequence(5'- 3')
退火温度 /℃
Anneal tempera-
ture
扩增条带 /数
Percentage of
polymorphic
bands
多态性条带 /数
Percentage of
polymorphic
bands
多态性百分率 /%
Percentage of poly-
morphic bands
UBC808 AGA GAG AGA GAG AGA GC 52. 3 8 8 100. 00
UBC809 AGA GAG AGA GAG AGA GG 53. 4 8 5 62. 50
UBC817 CAC ACA CAC ACA CAC AA 51 7 4 57. 14
UBC818 CAC ACA CAC ACA CAC AG 51 8 5 62. 50
UBC825 ACA CAC ACA CAC ACA CT 52. 3 8 7 87. 50
UBC826 ACA CAC ACA CAC ACA CC 58. 1 6 6 100. 00
UBC827 ACA CAC ACA CAC ACA CG 52. 3 7 6 85. 71
UBC828 TGT GTG TGT GTG TGT GA 51 5 3 60. 00
UBC830 TGT GTG TGT GTG TGT GG 54. 6 5 5 100. 00
UBC836 AGA GAG AGA GAG AGA GYA 51 9 8 88. 89
UBC841 GAG AGA GAG AGA GAG AYC 49. 9 6 5 83. 33
UBC846 CAC ACA CAC ACA CAC ART 49 7 5 71. 43
UBC847 CAC ACA CAC ACA CAC ARC 52. 3 8 7 87. 50
UBC848 CAC ACA CAC ACA CAC ARG 54. 6 7 6 85. 71
UBC849 GTG TGT GTG TGT GTG TYA 53. 4 9 8 88. 89
UBC851 GTG TGT GTG TGT GTG TYG 49. 9 7 7 100. 00
UBC856 ACA CAC ACA CAC ACA CYA 52. 3 11 10 90. 91
UBC857 ACA CAC ACA CAC ACA CYG 51 8 8 100. 00
UBC858 TGT GTG TGT GTG TGT GRT 49 5 3 60. 00
UBC866 CTC CTC CTC CTC CTC CTC 54. 6 5 4 80. 00
合计 144 120
平均 7. 2 6 83. 33
902第 2 期 罗 灿等:肾茶种质资源遗传多样性的 ISSR分析
2. 2 供试材料的遗传关系
2. 2. 1 ISSR遗传标记相似性分析 利用 20 条 ISSR标记引物对 87 份肾茶种质资源所获得的 144 条扩增
片段,通过软件 NTSYS2. 1 分析 87 份供试材料间的 Nei-Li 遗传相似系数 (GS 值) ,GS 值的变化范围在
0. 48 ~ 0. 98,平均值为 0. 73;从表 3 可见,在 87 份肾茶材料中,S29 与 S69 的遗传相似系数最小,为 0. 48,
表明 S29 与 S69 亲缘关系最远;81 份野生材料与 6 份栽培品种之间的遗传相似系数在 0. 48 ~ 0. 62 之间,
表明野生材料与栽培品种之间亲缘关系较远;6 份栽培种 S67,S68,S69,S70,S71,S72 之间的遗传相似系
数在 0. 88 ~ 0. 98 之间,表明 6 份栽培种之间亲缘关系较近;S69 与 S70 遗传相似系数最高,为 0. 98,表明
这 2 份材料遗传相似性最高,基因差异最小,亲缘关系最近。87 份肾茶遗传相似性分析结果表明,中国肾
茶资源存在丰富的遗传多样性。基于 ISSR数据计算的 87 份供试材料的相似系数详见附表。
表 3 基于 ISSR数据计算的 87 份供试材料的相似系数(部分)
Tab. 3 Genetic similarity of 87 C. spicatus accessions based on ISSR analysis
编号 S1 S2 S23 S24 S28 S29 S57 S58 S66 S67 S68 S69 S70 S71 S72
S1 1. 00
S2 0. 95 1. 00
S23 0. 86 0. 84 1. 00
S24 0. 67 0. 65 0. 63 1. 00
S28 0. 69 0. 68 0. 63 0. 84 1. 00
S29 0. 65 0. 64 0. 56 0. 78 0. 82 1. 00
S57 0. 72 0. 69 0. 63 0. 80 0. 82 0. 82 1. 00
S58 0. 81 0. 80 0. 74 0. 78 0. 80 0. 72 0. 81 1. 00
S66 0. 85 0. 84 0. 85 0. 67 0. 67 0. 60 0. 69 0. 79 1. 00
S67 0. 60 0. 56 0. 58 0. 58 0. 57 0. 50 0. 60 0. 59 0. 60 1. 00
S68 0. 60 0. 56 0. 58 0. 58 0. 60 0. 51 0. 63 0. 58 0. 62 0. 94 1. 00
S69 0. 63 0. 60 0. 58 0. 56 0. 56 0. 48 0. 56 0. 57 0. 65 0. 94 0. 92 1. 00
S70 0. 60 0. 58 0. 56 0. 56 0. 57 0. 50 0. 58 0. 58 0. 63 0. 93 0. 92 0. 98 1. 00
S71 0. 60 0. 57 0. 55 0. 58 0. 57 0. 53 0. 63 0. 60 0. 62 0. 93 0. 93 0. 92 0. 92 1. 00
S72 0. 58 0. 55 0. 54 0. 60 0. 62 0. 56 0. 66 0. 61 0. 60 0. 91 0. 92 0. 88 0. 88 0. 90 1. 00
2. 2. 2 肾茶遗传多样性的分子聚类 图 2 可见,当遗传相似系数为 0. 66 时,供试材料聚为 2 大类,第Ⅰ
大类包括 81 份材料,全部为海南野生种,它们之间的遗传相似系数变化范围为 0. 57 ~ 0. 95,表明野生材
料之间遗传多样性较丰富;第Ⅱ大类包含 6 份材料,全部都为栽培种,它们遗传相似系数在 0. 88 ~ 0. 98 之
间,说明它们之间的亲缘关系很近,但这 6 份材料来自不同地域,S67 和 S68 采自海南儋州热带植物园,
S69 和 S70 采自广西药用植物园,S71 与 S72 采自云南西双版纳植物园,可见肾茶的遗传多样性与生长的
地域没有较大的相关性,此聚类明显突出了野生材料与栽培材料之间遗传的差异性。在遗传相似系数为
0. 71 时,供试材料聚为 3 类,第Ⅰ大类可分为 2 个亚类,第 1 亚类包括 77 份材料,47 份采自霸王岭,2 份
采自五指山,分别为 S86 与 S87,28 份采自鹦哥岭;第 2 亚类包含 4 份材料,S12,S13,S14 及 S16 都为海南
霸王岭采集的野生资源,结果再次表明,其遗传聚类与地域没有严格相关性。
012 热 带 生 物 学 报 2015 年
3 讨 论
祁建明等[27]应用 ISSR 标记,对烟草属(Nicotiana)4 个种 30 份材料的遗传多样性进行分析,3 个野
生种的遗传相似系数在 0. 29 ~ 0. 52 之间,结果表明,ISSR很好地揭示出 3 个烟草野生种之间较大的遗传
差异性。本研究首次采用 ISSR对我国肾茶种质资源的遗传多样性进行分析,20 条 ISSR引物共产生 144 条
扩增条带,其中多态性条带 120条,多态率 83. 33%,87份材料间的遗传相似系数变化范围为 0. 48 ~0. 98,平
均 GS值为 0. 73,表明肾茶种质资源的遗传多样性比较丰富;这与刘伟等[28]利用 ISSR标记技术揭示西南
地区 45 份狗牙根之间的遗传多样性分析是可行的结论一致。海南野生肾茶材料间的遗传相似系数范围
为 0. 57 ~ 0. 95,表明野生材料具有较丰富的遗传多样性;栽培品种间遗传相似系数为 0. 88 ~ 0. 98,显示出
栽培种的遗传基础相对狭窄。究其原因,可能是栽培肾茶为同一品种生长在不同地域而已,也可能是同
112第 2 期 罗 灿等:肾茶种质资源遗传多样性的 ISSR分析
一品种引种到不同地方,再加上肾茶主要是无性繁殖为主,自然繁育比较低,因此基因交流较少,基因型
的改变较难,同时 6 份材料之间的叶形、叶缘、花色等形态特征都极其相似,这与表型的观察结果也是一
致的。
本试验利用 20 条引物能够将 87 份材料分开,并利用 UPGMA法将提供的材料聚成 2 大类,进一步证
实了 ISSR分子标记对肾茶资源遗传多样性分析是可行的。Chen 等[29]用 ISSR 分子标记对中国 92 个藕
莲(Nelumbo nucifera Gaertn.)进行遗传多样性分析,结果 92 个品种划分为 2 类,一类是大部分起源于我国
的地方品种,另一类是引种于美国的地方品种,这与肾茶遗传多样性分析的结果类似,显示出 ISSR 对肾
茶的遗传多样性分析是可行的。Joshi等[30]用 ISSR 标记研究稻(Oryza L.)的 42 个品种,包括 28 个野生
品种(9 个基因型)、1 个栽培种以及 3 个相关属的品种,发现在种间和种内均表现出很高的多态性,研
究结果表明,稻属是多途径进化的,这与肾茶遗传多样性分析结果比较类似,这为肾茶的种类鉴定提供了
依据。特别值得注意的是 87 份供试材料中不同区域栽培品种与野生材料之间的遗传差异性较大,野生
材料之间同样表现为遗传差异性较大,可见肾茶的遗传聚类与其地理来源没有严格的一致性,这与刘建
乐等[31]对割手密种质资源遗传多样性研究结果一致。
本研究用 ISSR标记技术对 87份肾茶材料进行遗传聚类与遗传相似性分析,为了解中国肾茶资源地理
起源和肾茶种类鉴定提供了依据。近年来,肾茶虽有大量引种栽培,但随着人们对野生肾茶资源的破坏,海
南野生资源已处于濒危状态,再加上海南环境的影响,一些野生居群正在初步缩小甚至消失,这对于肾茶种
质资源的研究是非常不利的,因此,加强肾茶野生资源的保护刻不容缓,这也是本研究的意义所在。
致谢:海南大学农学院戚山江、赵原森同学、霸王岭自然保护区王进强老师在实验过程中给予了帮助,特此致谢!
参考文献:
[1]柳丹萍,黄荣桂. 肾茶研究新进展[J]. 海峡药学,2013,25(11) :56 - 60.
[2]中国科学院中国植物志编辑委员会.中国植物志(第六十六卷) [M]. 北京:科学出版社,1999:574 - 577.
[3]罗关兴,娄尚椿. 珍贵药用保健植物———肾茶[J]. 广西热带农业,2004(6) :31 - 32.
[4]李芳洲,盛国荣,许金星,等. 肾茶(大花直管草)的引种栽培和初步临床试验报告[J]. 亚热带植物通讯,1980(1) :19 -24.
[5]张平,李伯儒. 肾茶移栽密度、荫蔽度和追肥量的研究初报[J]. 热带作物研究,1997(3) :39 - 41.
[6]陈建白,白旭华,蒋桂芝,等. 肾茶开发利用的初步研究[J]. 云南热作科技,1988,21(3) :6 - 8.
[7]许娜,许旭东,杨峻山,等. 猫须草的研究进展[J]. 中草药,2010,41(5) :848 - 851.
[8]高南南,陈慧珍,田泽,等. 肾茶药理作用的研究[J]. 中草药,1996,27(10) :615 - 615.
[9]蔡华芳,寿燕,汪菁菁,等. 肾茶的药理作用初探[J]. 中药材,1997,20(1) :38 - 40.
[10]李月婷,黄荣桂,郑兴中,等. 肾茶的研究进展[J]. 中国中西医结合杂志,2002,22(6) :470 - 472.
[11]颜文祝,吕于谋,沈文通,等. 肾茶主要化学成份初步试验[J]. 海峡药学,1995,7(3) :3 - 4.
[12]陈伊蕾,谭昌恒,谭俊杰,等. 肾茶的化学和药理研究进展[J]. 天然产物研究与开发,2009,21(5) :885 - 891.
[13]DOLEKOV I,RROV L,GRZ J,et al. Antiproliferative and antiangiogenic effects of flavone eupatorin,an active con-
stituent of chloroform extract of Orthosiphon stamineus leaves[J]. Fitoterapia,2012,83(6) :1000 - 1007.
[14]AMZAD H M,ISMAIL Z. Isolation and characterization of triterpenes from the leaves of Orthosiphon stamineus[J],Arabian
Journal of Chemistry,2013,6:295 - 298.
[15]ZIETKIEWICZ E,RAFALSKI A,LABUDA D. Genome fingerprinting by simple sequence repeat (SSR)-anchored polymerase
chain reaction amplification[J]. Genomics,1994,20:176 - 183.
[16]GUPTA M,CHYI Y S,ROMERO-SEVERSON J,et al. Amplification of DNA makers from evolutionarily diverse genomes u-
sing single primers of simple-sequence repeats[J]. Theoretical and Applied Genetics,1994,89:998 - 1006.
[17]NAGAOKA T,OGIHARA Y. Applicability of inter-simple sequence repeat polymorphisms in wheat for use as DNA markers in
comparison to RFLP and RAPD marker[J]. Theoretical and Applied Genetics,1997,94:597 - 602.
[18]TSUMURA Y,OHBA K,TRAUSS S H. Diversity and inheritance of inter-simple sequence repeat polymorphisms in Douglas-
fir (Pseudotsuga menziesii)and sugi(Cryptomeria japonica) [J]. Theoretical and Applied Genetics,1996,92(1) :40 - 45.
[19]QI J M,ZHOU D X,WU W R,et al. Applicat ion of ISSR technology in genetic diversity detection of Jute[J]. Chinese Jour-
nal of Applied Ecology,2003,14(9) :1473 - 1477.
[20]WANG X Y,CHEN P D,QI Z J,et al. Preliminary study of ISSR marker technique on wheat fingerprinting[J]. Journal of Ag-
212 热 带 生 物 学 报 2015 年
ricultural Biotechnology ,2001,9(3) :261 - 263.
[21]GE Y Q,QIU Y X,DING B Y,et al. An ISSR analysis on population genetic diversity of the relict plant ginkgo Biloba[J].
Biodiversity Sci,2003,11(4) :276 - 287.
[22]王英,庄南生,高和琼,等. 甘蔗种质遗传基础的 ISSR分析[J]. 湖南农业大学学报(自然科学版),2007,33(Z1):176 -183.
[23]BEHERA T K,SINGH A K,STAUB J E. Comparative analysis of genetic diversity in Indian bitter gourd (Momordica charantia
L. )using RAPD and ISSR markers for developing crop improvement strategies[J]. Scientia Horticulturae,2008,15:209 -
217.
[24]广东省植物研究所编委会.海南植物志(第四卷上) [M].北京:科学出版社,1977:56 - 57.
[25]CHAUDHRY B,YASMEEN A,HUSNAIN T,et al. Mini-scale Genomic DNA Extraction from Cotton[J]. Plant Molecular
Biology Repoter,1999,17(3) :280 - 280.
[26]VROHBI I,HARVENGT L,CHANDELIER A,et al. Improved RAPD amplification of recalcitrant plant DNA by the use of
activated charcoal during DNA extraction[J]. Plant Breed,1996,115:205 - 206.
[27]祁建民,王涛,陈顺辉,等. 部分烟草种质遗传多样性与亲缘关系的 ISSR标记分析[J]. 作物学报,2006,32(3):373 -378.
[28]刘伟,张新全,李芳,等. 西南区野生狗牙根遗传多样性的 ISSR标记与地理来源分析[J]. 草业学报,2007,16(3):55 -61.
[29]CHEN Y Y,ZHOU R C,LIN X D,et al. ISSR analys is of genetic diversity in sacred lotus cultivars[J]. Aquatic Botany,
2008,89:311 - 316.
[30]JOSHI S P,GUPTA V S,AGGARWAL R K,et al. Genetic diversity and phylogenetic relationship as revealed by inter-simple
sequence repeat (ISSR )polymorphism in the genus Oryza[J]. Theoretical and Applied Genetics,2000,100(8) :1311 -
1320.
[31]刘建乐,白昌军,严琳玲,等. 割手密种质资源遗传多样性的 ISSR分析[J]. 热带作物学报,2014,35(1) :68 - 73.
Genetic Diversity in Clerodendranthus spicatus (Thunb.)
Based on ISSR Markers
LUO Can1,YU Xudong1,2,3,WU Fanhua2,GAO Xin3,WANG Tongxin2
(1. College of Horticulture and Landscape Architecture,Hainan University,Haikou,570228;2. Laboratory for Development and Utilization of
Tropical Rare Plants Resources,College of Agronomy,Hainan University,Haikou,570228;3. College of Applied Science and Technology,Hainan
University,Danzhou,571737,China)
Abstract:In order to analyze the genetic diversity of Clerodendranthus spicatus (Thunb.)C. Y. Wu. in China,a
total of 87 accessions of C. spicatus (Thunb.)C. Y. Wu were analyzed by using ISSR markers. One hundred of
ISSR primers were designed to produce amplification products of the accessions,among which 20 primers pro-
duced distinct amplification products. The 20 primers yielded a total of 144 bands in the 87 accessions,of which
120 bands (83. 3%)were polymorphic (6. 0 polymorphic bands per primer). The ISSR-based genetic similarity
(GS)values of the 87 C. spicatus accessions ranged from 0. 48 to 0. 98,but the GS values of six accessions of
cultivars ranged from 0. 88 - 0. 98. This result indicated that the genetic basis was comparatively narrow among
the cultivars. Moreover,the 87 accessions were clustered into two groups by UPGMA,among which the cultivars
were classified into one group and the wild species into another group. It is concluded that the wild germplasm
resources of C. spicatus in China are more genetically diverse and hence have a great potential for further exploi-
tation and utilization.
Key words:Clerodendranthus spicatus (Thunb.)C. Y. Wu.;CTAB methods;ISSR;genetic diversity
312第 2 期 罗 灿等:肾茶种质资源遗传多样性的 ISSR分析
附表 基于 ISSR数据计算的 87 份供试材料的相似系数
Schedule Genetic similarity of 87 C. spicatus accessions based on ISSR analysis
编号 S1 S2 S3 S4 S5 S6 S7 S8 S9 S10 S11 S12 S13 S14 S15
S1 1. 00
S1 1. 00
S2 0. 95 1. 00
S3 0. 85 0. 85 1. 00
S4 0. 79 0. 77 0. 81 1. 00
S5 0. 84 0. 85 0. 88 0. 76 1. 00
S6 0. 83 0. 83 0. 83 0. 75 0. 83 1. 00
S7 0. 87 0. 89 0. 90 0. 74 0. 94 0. 87 1. 00
S8 0. 78 0. 76 0. 80 0. 85 0. 75 0. 76 0. 76 1. 00
S9 0. 85 0. 85 0. 81 0. 78 0. 80 0. 85 0. 84 0. 77 1. 00
S10 0. 82 0. 84 0. 81 0. 76 0. 81 0. 85 0. 84 0. 77 0. 90 1. 00
S11 0. 81 0. 83 0. 79 0. 76 0. 78 0. 86 0. 84 0. 76 0. 88 0. 93 1. 00
S12 0. 65 0. 64 0. 66 0. 73 0. 61 0. 70 0. 63 0. 74 0. 72 0. 70 0. 72 1. 00
S13 0. 73 0. 72 0. 76 0. 76 0. 71 0. 76 0. 72 0. 75 0. 80 0. 73 0. 77 0. 82 1. 00
S14 0. 69 0. 69 0. 70 0. 72 0. 65 0. 73 0. 68 0. 72 0. 78 0. 78 0. 80 0. 83 0. 83 1. 00
S15 0. 77 0. 78 0. 78 0. 74 0. 79 0. 76 0. 81 0. 75 0. 81 0. 84 0. 85 0. 64 0. 75 0. 78 1. 00
S16 0. 65 0. 65 0. 66 0. 66 0. 61 0. 73 0. 64 0. 68 0. 72 0. 70 0. 73 0. 85 0. 81 0. 86 0. 68
S17 0. 74 0. 74 0. 78 0. 75 0. 76 0. 72 0. 76 0. 73 0. 76 0. 76 0. 75 0. 58 0. 70 0. 69 0. 76
S18 0. 72 0. 70 0. 79 0. 85 0. 72 0. 71 0. 70 0. 83 0. 71 0. 72 0. 72 0. 69 0. 77 0. 70 0. 74
S19 0. 82 0. 78 0. 79 0. 69 0. 81 0. 78 0. 84 0. 74 0. 81 0. 83 0. 82 0. 58 0. 67 0. 69 0. 83
S20 0. 85 0. 88 0. 83 0. 77 0. 83 0. 81 0. 86 0. 76 0. 88 0. 91 0. 87 0. 64 0. 69 0. 72 0. 81
S21 0. 78 0. 78 0. 78 0. 81 0. 77 0. 75 0. 80 0. 78 0. 85 0. 88 0. 85 0. 63 0. 73 0. 73 0. 87
S22 0. 76 0. 74 0. 81 0. 79 0. 76 0. 81 0. 78 0. 80 0. 82 0. 83 0. 81 0. 72 0. 78 0. 72 0. 76
S23 0. 86 0. 84 0. 82 0. 71 0. 83 0. 81 0. 84 0. 72 0. 82 0. 85 0. 81 0. 59 0. 65 0. 67 0. 74
S24 0. 67 0. 65 0. 72 0. 85 0. 62 0. 65 0. 62 0. 78 0. 68 0. 68 0. 65 0. 70 0. 72 0. 69 0. 65
S25 0. 84 0. 83 0. 84 0. 72 0. 82 0. 77 0. 83 0. 76 0. 78 0. 83 0. 78 0. 60 0. 69 0. 69 0. 78
S26 0. 82 0. 78 0. 81 0. 76 0. 77 0. 78 0. 80 0. 78 0. 82 0. 86 0. 81 0. 65 0. 70 0. 72 0. 77
S27 0. 76 0. 75 0. 80 0. 84 0. 75 0. 77 0. 78 0. 81 0. 78 0. 81 0. 78 0. 69 0. 74 0. 71 0. 76
S28 0. 69 0. 68 0. 76 0. 81 0. 67 0. 69 0. 67 0. 82 0. 66 0. 70 0. 67 0. 76 0. 72 0. 74 0. 68
S29 0. 65 0. 64 0. 67 0. 73 0. 63 0. 65 0. 61 0. 75 0. 65 0. 62 0. 62 0. 76 0. 72 0. 67 0. 63
S30 0. 72 0. 73 0. 78 0. 72 0. 74 0. 76 0. 76 0. 81 0. 76 0. 82 0. 76 0. 74 0. 76 0. 74 0. 76
S31 0. 75 0. 74 0. 81 0. 71 0. 80 0. 79 0. 80 0. 77 0. 75 0. 81 0. 76 0. 67 0. 74 0. 72 0. 77
S32 0. 80 0. 82 0. 83 0. 76 0. 74 0. 78 0. 79 0. 76 0. 80 0. 87 0. 84 0. 69 0. 69 0. 74 0. 81
412 热 带 生 物 学 报 2015 年
续附表 Renew schedule
编号 S1 S2 S3 S4 S5 S6 S7 S8 S9 S10 S11 S12 S13 S14 S15
S33 0. 79 0. 81 0. 78 0. 74 0. 76 0. 76 0. 81 0. 73 0. 79 0. 79 0. 78 0. 63 0. 70 0. 66 0. 74
S34 0. 79 0. 78 0. 76 0. 78 0. 76 0. 75 0. 80 0. 74 0. 82 0. 82 0. 78 0. 66 0. 72 0. 69 0. 76
S35 0. 80 0. 79 0. 80 0. 77 0. 76 0. 77 0. 81 0. 75 0. 78 0. 80 0. 77 0. 63 0. 71 0. 65 0. 75
S36 0. 81 0. 83 0. 81 0. 76 0. 82 0. 77 0. 83 0. 76 0. 81 0. 84 0. 80 0. 63 0. 67 0. 67 0. 76
S37 0. 82 0. 84 0. 83 0. 76 0. 78 0. 79 0. 83 0. 78 0. 81 0. 82 0. 81 0. 62 0. 67 0. 70 0. 78
S38 0. 68 0. 67 0. 69 0. 69 0. 70 0. 64 0. 66 0. 67 0. 65 0. 69 0. 64 0. 63 0. 62 0. 63 0. 67
S39 0. 79 0. 81 0. 83 0. 76 0. 78 0. 75 0. 78 0. 78 0. 79 0. 82 0. 76 0. 67 0. 73 0. 73 0. 74
S40 0. 82 0. 83 0. 85 0. 78 0. 83 0. 79 0. 83 0. 78 0. 82 0. 85 0. 79 0. 67 0. 73 0. 73 0. 76
S41 0. 78 0. 81 0. 82 0. 79 0. 80 0. 78 0. 80 0. 78 0. 81 0. 86 0. 79 0. 66 0. 72 0. 73 0. 80
S42 0. 75 0. 78 0. 82 0. 78 0. 77 0. 78 0. 76 0. 77 0. 78 0. 83 0. 81 0. 67 0. 73 0. 72 0. 80
S43 0. 78 0. 78 0. 83 0. 80 0. 79 0. 78 0. 79 0. 78 0. 78 0. 81 0. 78 0. 67 0. 75 0. 72 0. 76
S44 0. 74 0. 74 0. 75 0. 72 0. 72 0. 76 0. 73 0. 72 0. 75 0. 78 0. 72 0. 80 0. 77 0. 77 0. 70
S45 0. 72 0. 74 0. 78 0. 72 0. 73 0. 79 0. 77 0. 70 0. 75 0. 81 0. 79 0. 70 0. 67 0. 74 0. 74
S46 0. 73 0. 75 0. 78 0. 70 0. 71 0. 80 0. 76 0. 71 0. 78 0. 81 0. 78 0. 71 0. 69 0. 74 0. 74
S47 0. 66 0. 67 0. 70 0. 70 0. 67 0. 67 0. 68 0. 68 0. 65 0. 70 0. 65 0. 68 0. 67 0. 68 0. 67
S48 0. 78 0. 74 0. 81 0. 75 0. 74 0. 75 0. 78 0. 76 0. 76 0. 81 0. 81 0. 62 0. 66 0. 69 0. 76
S49 0. 83 0. 81 0. 83 0. 85 0. 82 0. 78 0. 82 0. 82 0. 81 0. 80 0. 80 0. 67 0. 72 0. 74 0. 82
S50 0. 83 0. 83 0. 78 0. 74 0. 80 0. 79 0. 83 0. 72 0. 79 0. 76 0. 78 0. 58 0. 65 0. 65 0. 74
S51 0. 78 0. 81 0. 78 0. 76 0. 75 0. 81 0. 79 0. 76 0. 80 0. 85 0. 84 0. 65 0. 71 0. 74 0. 82
S52 0. 78 0. 78 0. 83 0. 79 0. 78 0. 83 0. 81 0. 80 0. 78 0. 79 0. 76 0. 66 0. 72 0. 72 0. 74
S53 0. 78 0. 78 0. 81 0. 84 0. 75 0. 81 0. 76 0. 79 0. 76 0. 74 0. 74 0. 68 0. 76 0. 71 0. 71
S54 0. 78 0. 77 0. 78 0. 83 0. 76 0. 79 0. 77 0. 83 0. 76 0. 75 0. 76 0. 69 0. 76 0. 70 0. 74
S55 0. 77 0. 75 0. 77 0. 83 0. 76 0. 76 0. 74 0. 79 0. 76 0. 74 0. 73 0. 68 0. 75 0. 69 0. 74
S56 0. 81 0. 83 0. 78 0. 74 0. 79 0. 78 0. 81 0. 75 0. 78 0. 78 0. 77 0. 61 0. 67 0. 67 0. 71
S57 0. 72 0. 69 0. 72 0. 80 0. 68 0. 72 0. 67 0. 79 0. 69 0. 69 0. 69 0. 72 0. 71 0. 68 0. 68
S58 0. 81 0. 80 0. 82 0. 79 0. 76 0. 79 0. 78 0. 85 0. 79 0. 79 0. 76 0. 67 0. 74 0. 72 0. 76
S59 0. 84 0. 82 0. 81 0. 80 0. 78 0. 88 0. 79 0. 79 0. 81 0. 78 0. 80 0. 69 0. 76 0. 69 0. 74
S60 0. 71 0. 69 0. 72 0. 75 0. 66 0. 65 0. 66 0. 74 0. 69 0. 68 0. 67 0. 74 0. 74 0. 67 0. 69
S61 0. 82 0. 83 0. 79 0. 69 0. 81 0. 82 0. 83 0. 74 0. 81 0. 79 0. 75 0. 66 0. 72 0. 66 0. 72
S62 0. 79 0. 77 0. 81 0. 79 0. 78 0. 88 0. 81 0. 77 0. 83 0. 81 0. 82 0. 67 0. 74 0. 72 0. 76
S63 0. 81 0. 80 0. 82 0. 76 0. 81 0. 82 0. 83 0. 81 0. 81 0. 82 0. 81 0. 67 0. 73 0. 73 0. 78
S64 0. 85 0. 85 0. 84 0. 80 0. 82 0. 84 0. 85 0. 85 0. 84 0. 83 0. 80 0. 67 0. 72 0. 71 0. 78
S65 0. 81 0. 81 0. 84 0. 78 0. 83 0. 81 0. 83 0. 83 0. 80 0. 81 0. 78 0. 67 0. 74 0. 74 0. 81
512第 2 期 罗 灿等:肾茶种质资源遗传多样性的 ISSR分析
续附表 Renew schedule
编号 S1 S2 S3 S4 S5 S6 S7 S8 S9 S10 S11 S12 S13 S14 S15
S66 0. 85 0. 84 0. 79 0. 75 0. 81 0. 78 0. 83 0. 78 0. 79 0. 78 0. 75 0. 62 0. 69 0. 69 0. 76
S67 0. 60 0. 56 0. 52 0. 58 0. 54 0. 53 0. 57 0. 56 0. 55 0. 55 0. 53 0. 53 0. 53 0. 56 0. 54
S68 0. 60 0. 56 0. 53 0. 59 0. 56 0. 55 0. 57 0. 60 0. 55 0. 53 0. 52 0. 57 0. 56 0. 57 0. 56
S69 0. 63 0. 60 0. 54 0. 58 0. 56 0. 56 0. 58 0. 59 0. 58 0. 58 0. 56 0. 56 0. 55 0. 59 0. 58
S70 0. 60 0. 58 0. 53 0. 58 0. 56 0. 55 0. 57 0. 58 0. 56 0. 56 0. 55 0. 57 0. 56 0. 60 0. 58
S71 0. 60 0. 57 0. 53 0. 59 0. 57 0. 58 0. 60 0. 61 0. 59 0. 58 0. 56 0. 58 0. 57 0. 58 0. 58
S72 0. 58 0. 55 0. 51 0. 60 0. 52 0. 54 0. 55 0. 60 0. 54 0. 53 0. 51 0. 58 0. 56 0. 58 0. 52
S73 0. 83 0. 79 0. 77 0. 77 0. 78 0. 76 0. 78 0. 81 0. 80 0. 81 0. 77 0. 68 0. 74 0. 74 0. 76
S74 0. 77 0. 78 0. 77 0. 77 0. 76 0. 73 0. 76 0. 78 0. 77 0. 80 0. 76 0. 64 0. 69 0. 68 0. 74
S75 0. 76 0. 74 0. 78 0. 80 0. 79 0. 78 0. 76 0. 81 0. 81 0. 78 0. 74 0. 68 0. 76 0. 72 0. 72
S76 0. 74 0. 73 0. 78 0. 78 0. 74 0. 75 0. 74 0. 78 0. 76 0. 76 0. 74 0. 73 0. 81 0. 74 0. 72
S77 0. 72 0. 72 0. 76 0. 72 0. 74 0. 81 0. 76 0. 76 0. 78 0. 78 0. 78 0. 80 0. 83 0. 80 0. 70
S78 0. 74 0. 73 0. 79 0. 76 0. 77 0. 79 0. 76 0. 80 0. 74 0. 76 0. 72 0. 67 0. 69 0. 69 0. 69
S79 0. 75 0. 76 0. 81 0. 78 0. 74 0. 78 0. 74 0. 81 0. 74 0. 76 0. 74 0. 70 0. 70 0. 67 0. 69
S80 0. 78 0. 79 0. 83 0. 80 0. 76 0. 80 0. 78 0. 81 0. 77 0. 80 0. 77 0. 64 0. 69 0. 65 0. 72
S81 0. 75 0. 73 0. 79 0. 76 0. 74 0. 76 0. 74 0. 76 0. 72 0. 72 0. 69 0. 65 0. 70 0. 67 0. 67
S82 0. 72 0. 73 0. 79 0. 81 0. 72 0. 75 0. 70 0. 77 0. 72 0. 75 0. 74 0. 73 0. 72 0. 72 0. 70
S83 0. 78 0. 80 0. 81 0. 75 0. 77 0. 76 0. 77 0. 78 0. 74 0. 78 0. 74 0. 65 0. 65 0. 66 0. 70
S84 0. 83 0. 83 0. 83 0. 76 0. 79 0. 81 0. 83 0. 76 0. 81 0. 83 0. 83 0. 67 0. 74 0. 71 0. 78
S85 0. 82 0. 81 0. 83 0. 81 0. 81 0. 83 0. 81 0. 83 0. 79 0. 82 0. 79 0. 70 0. 74 0. 70 0. 72
S86 0. 81 0. 81 0. 84 0. 83 0. 82 0. 83 0. 83 0. 78 0. 85 0. 88 0. 83 0. 67 0. 72 0. 72 0. 78
S87 0. 83 0. 81 0. 85 0. 81 0. 81 0. 83 0. 83 0. 78 0. 85 0. 86 0. 81 0. 70 0. 74 0. 74 0. 77
编号 S16 S17 S18 S19 S20 S21 S22 S23 S24 S25 S26 S27 S28 S29 S30
S16 1. 00
S17 0. 66 1. 00
S18 0. 67 0. 82 1. 00
S19 0. 59 0. 81 0. 74 1. 00
S20 0. 63 0. 76 0. 72 0. 85 1. 00
S21 0. 65 0. 81 0. 78 0. 83 0. 87 1. 00
S22 0. 70 0. 74 0. 82 0. 78 0. 84 0. 83 1. 00
S23 0. 60 0. 75 0. 68 0. 89 0. 90 0. 81 0. 78 1. 00
612 热 带 生 物 学 报 2015 年
续附表 Renew Schedule
编号 S16 S17 S18 S19 S20 S21 S22 S23 S24 S25 S26 S27 S28 S29 S30
S24 0. 69 0. 72 0. 83 0. 63 0. 67 0. 74 0. 79 0. 63 1. 00
S25 0. 60 0. 81 0. 74 0. 91 0. 88 0. 81 0. 74 0. 91 0. 65 1. 00
S26 0. 62 0. 81 0. 74 0. 88 0. 87 0. 83 0. 81 0. 86 0. 71 0. 91 1. 00
S27 0. 65 0. 80 0. 81 0. 80 0. 82 0. 85 0. 83 0. 76 0. 80 0. 81 0. 85 1. 00
S28 0. 76 0. 76 0. 85 0. 66 0. 67 0. 73 0. 76 0. 63 0. 84 0. 69 0. 70 0. 82 1. 00
S29 0. 69 0. 67 0. 76 0. 58 0. 58 0. 66 0. 66 0. 56 0. 78 0. 61 0. 58 0. 71 0. 82 1. 00
S30 0. 70 0. 71 0. 78 0. 75 0. 76 0. 81 0. 82 0. 72 0. 72 0. 77 0. 79 0. 78 0. 80 0. 73 1. 00
S31 0. 67 0. 72 0. 78 0. 82 0. 77 0. 78 0. 83 0. 76 0. 69 0. 78 0. 78 0. 77 0. 77 0. 69 0. 92
S32 0. 68 0. 76 0. 74 0. 78 0. 82 0. 81 0. 77 0. 78 0. 69 0. 81 0. 83 0. 79 0. 75 0. 65 0. 81
S33 0. 66 0. 75 0. 75 0. 78 0. 81 0. 81 0. 81 0. 78 0. 69 0. 78 0. 78 0. 83 0. 74 0. 65 0. 75
S34 0. 65 0. 74 0. 74 0. 76 0. 83 0. 85 0. 82 0. 78 0. 69 0. 77 0. 82 0. 83 0. 73 0. 65 0. 76
S35 0. 67 0. 76 0. 77 0. 77 0. 79 0. 80 0. 81 0. 77 0. 72 0. 76 0. 80 0. 83 0. 76 0. 67 0. 76
S36 0. 60 0. 76 0. 74 0. 81 0. 89 0. 81 0. 78 0. 83 0. 63 0. 86 0. 84 0. 81 0. 71 0. 61 0. 77
S37 0. 60 0. 75 0. 74 0. 82 0. 87 0. 79 0. 81 0. 82 0. 68 0. 84 0. 85 0. 81 0. 72 0. 62 0. 74
S38 0. 60 0. 75 0. 74 0. 69 0. 70 0. 72 0. 67 0. 68 0. 68 0. 76 0. 74 0. 76 0. 78 0. 73 0. 71
S39 0. 67 0. 79 0. 78 0. 76 0. 84 0. 78 0. 74 0. 67 0. 79 0. 78 0. 76 0. 84 0. 78 0. 67 0. 78
S40 0. 67 0. 79 0. 76 0. 78 0. 85 0. 81 0. 79 0. 81 0. 68 0. 85 0. 85 0. 83 0. 76 0. 66 0. 78
S41 0. 65 0. 78 0. 76 0. 81 0. 81 0. 85 0. 78 0. 79 0. 72 0. 84 0. 83 0. 83 0. 76 0. 70 0. 83
S42 0. 67 0. 79 0. 79 0. 79 0. 80 0. 79 0. 78 0. 76 0. 72 0. 83 0. 82 0. 81 0. 78 0. 67 0. 79
S43 0. 64 0. 76 0. 80 0. 80 0. 82 0. 81 0. 81 0. 78 0. 73 0. 82 0. 83 0. 85 0. 76 0. 68 0. 77
S44 0. 78 0. 69 0. 71 0. 69 0. 74 0. 72 0. 79 0. 72 0. 69 0. 73 0. 76 0. 78 0. 77 0. 69 0. 81
S45 0. 72 0. 72 0. 68 0. 72 0. 76 0. 78 0. 74 0. 75 0. 67 0. 74 0. 76 0. 80 0. 76 0. 69 0. 75
S46 0. 71 0. 69 0. 66 0. 72 0. 78 0. 77 0. 76 0. 74 0. 65 0. 74 0. 77 0. 76 0. 71 0. 67 0. 77
S47 0. 69 0. 67 0. 73 0. 66 0. 65 0. 70 0. 72 0. 63 0. 77 0. 68 0. 69 0. 72 0. 81 0. 76 0. 76
S48 0. 65 0. 79 0. 78 0. 83 0. 83 0. 82 0. 78 0. 82 0. 68 0. 85 0. 85 0. 83 0. 74 0. 66 0. 74
S49 0. 68 0. 78 0. 80 0. 77 0. 81 0. 81 0. 78 0. 76 0. 73 0. 78 0. 76 0. 79 0. 76 0. 74 0. 70
S50 0. 58 0. 72 0. 68 0. 81 0. 81 0. 74 0. 69 0. 85 0. 61 0. 83 0. 76 0. 74 0. 62 0. 59 0. 65
S51 0. 67 0. 77 0. 76 0. 83 0. 82 0. 88 0. 77 0. 80 0. 72 0. 81 0. 80 0. 83 0. 78 0. 69 0. 81
S52 0. 67 0. 83 0. 82 0. 79 0. 78 0. 79 0. 79 0. 78 0. 74 0. 80 0. 81 0. 84 0. 80 0. 69 0. 76
S53 0. 67 0. 76 0. 84 0. 72 0. 76 0. 78 0. 81 0. 73 0. 80 0. 74 0. 73 0. 81 0. 81 0. 74 0. 73
S54 0. 67 0. 79 0. 83 0. 74 0. 76 0. 78 0. 81 0. 71 0. 76 0. 73 0. 72 0. 81 0. 81 0. 74 0. 75
S55 0. 67 0. 78 0. 84 0. 72 0. 74 0. 76 0. 80 0. 72 0. 77 0. 72 0. 73 0. 79 0. 81 0. 75 0. 78
712第 2 期 罗 灿等:肾茶种质资源遗传多样性的 ISSR分析
续附表 Renew Schedule
编号 S16 S17 S18 S19 S20 S21 S22 S23 S24 S25 S26 S27 S28 S29 S30
S56 0. 64 0. 73 0. 73 0. 76 0. 79 0. 74 0. 73 0. 78 0. 66 0. 76 0. 76 0. 76 0. 69 0. 60 0. 70
S57 0. 65 0. 73 0. 80 0. 67 0. 67 0. 72 0. 74 0. 63 0. 80 0. 65 0. 67 0. 74 0. 82 0. 82 0. 72
S58 0. 69 0. 79 0. 83 0. 75 0. 77 0. 76 0. 79 0. 74 0. 78 0. 76 0. 79 0. 80 0. 80 0. 72 0. 78
S59 0. 69 0. 73 0. 74 0. 74 0. 76 0. 72 0. 76 0. 76 0. 69 0. 75 0. 78 0. 72 0. 71 0. 64 0. 74
S60 0. 66 0. 69 0. 78 0. 67 0. 66 0. 72 0. 74 0. 64 0. 74 0. 67 0. 68 0. 74 0. 78 0. 80 0. 74
S61 0. 65 0. 72 0. 69 0. 81 0. 77 0. 72 0. 75 0. 83 0. 64 0. 80 0. 78 0. 73 0. 66 0. 63 0. 75
S62 0. 72 0. 74 0. 76 0. 76 0. 78 0. 79 0. 81 0. 79 0. 72 0. 76 0. 78 0. 81 0. 73 0. 65 0. 72
S63 0. 67 0. 76 0. 78 0. 82 0. 83 0. 78 0. 82 0. 81 0. 71 0. 81 0. 82 0. 83 0. 73 0. 65 0. 76
S64 0. 67 0. 74 0. 77 0. 78 0. 83 0. 78 0. 81 0. 80 0. 70 0. 79 0. 81 0. 81 0. 72 0. 65 0. 77
S65 0. 68 0. 74 0. 77 0. 80 0. 82 0. 77 0. 81 0. 78 0. 70 0. 79 0. 80 0. 81 0. 74 0. 67 0. 78
S66 0. 62 0. 71 0. 74 0. 82 0. 84 0. 76 0. 76 0. 85 0. 67 0. 84 0. 79 0. 77 0. 67 0. 60 0. 75
S67 0. 50 0. 53 0. 53 0. 62 0. 58 0. 56 0. 58 0. 58 0. 58 0. 58 0. 60 0. 61 0. 57 0. 50 0. 55
S68 0. 53 0. 52 0. 56 0. 62 0. 57 0. 55 0. 60 0. 58 0. 58 0. 58 0. 59 0. 61 0. 60 0. 51 0. 58
S69 0. 53 0. 51 0. 53 0. 61 0. 60 0. 57 0. 60 0. 58 0. 56 0. 60 0. 61 0. 59 0. 56 0. 48 0. 60
S70 0. 54 0. 52 0. 53 0. 60 0. 58 0. 55 0. 59 0. 56 0. 56 0. 58 0. 59 0. 58 0. 57 0. 50 0. 58
S71 0. 54 0. 52 0. 55 0. 60 0. 58 0. 55 0. 60 0. 55 0. 58 0. 56 0. 60 0. 60 0. 57 0. 53 0. 59
S72 0. 53 0. 54 0. 57 0. 57 0. 56 0. 56 0. 60 0. 54 0. 60 0. 55 0. 57 0. 62 0. 62 0. 56 0. 57
S73 0. 68 0. 74 0. 77 0. 78 0. 79 0. 81 0. 76 0. 77 0. 70 0. 81 0. 80 0. 79 0. 78 0. 71 0. 81
S74 0. 64 0. 78 0. 78 0. 77 0. 79 0. 80 0. 76 0. 77 0. 72 0. 82 0. 78 0. 79 0. 76 0. 67 0. 80
S75 0. 67 0. 76 0. 80 0. 76 0. 78 0. 77 0. 85 0. 76 0. 77 0. 76 0. 80 0. 76 0. 75 0. 67 0. 84
S76 0. 73 0. 79 0. 88 0. 72 0. 73 0. 79 0. 82 0. 69 0. 78 0. 74 0. 74 0. 84 0. 87 0. 80 0. 86
S77 0. 80 0. 69 0. 75 0. 68 0. 70 0. 69 0. 79 0. 68 0. 71 0. 69 0. 69 0. 76 0. 78 0. 76 0. 85
S78 0. 67 0. 78 0. 78 0. 75 0. 74 0. 75 0. 78 0. 75 0. 72 0. 77 0. 75 0. 80 0. 81 0. 72 0. 81
S79 0. 69 0. 75 0. 78 0. 71 0. 74 0. 72 0. 75 0. 69 0. 72 0. 73 0. 72 0. 77 0. 83 0. 74 0. 78
S80 0. 63 0. 77 0. 81 0. 76 0. 79 0. 77 0. 78 0. 77 0. 74 0. 79 0. 80 0. 79 0. 74 0. 65 0. 76
S81 0. 67 0. 75 0. 82 0. 71 0. 70 0. 69 0. 78 0. 72 0. 74 0. 72 0. 72 0. 74 0. 77 0. 72 0. 78
S82 0. 74 0. 76 0. 83 0. 68 0. 70 0. 75 0. 76 0. 68 0. 78 0. 70 0. 71 0. 78 0. 87 0. 74 0. 76
S83 0. 65 0. 74 0. 75 0. 74 0. 76 0. 74 0. 74 0. 75 0. 68 0. 77 0. 75 0. 76 0. 74 0. 67 0. 76
S84 0. 67 0. 77 0. 78 0. 78 0. 81 0. 77 0. 80 0. 77 0. 69 0. 79 0. 80 0. 79 0. 71 0. 67 0. 80
S85 0. 66 0. 76 0. 82 0. 78 0. 80 0. 78 0. 83 0. 79 0. 72 0. 80 0. 81 0. 81 0. 74 0. 69 0. 79
S86 0. 64 0. 76 0. 77 0. 81 0. 86 0. 83 0. 81 0. 83 0. 72 0. 83 0. 85 0. 85 0. 72 0. 64 0. 80
S87 0. 67 0. 69 0. 75 0. 79 0. 84 0. 79 0. 83 0. 82 0. 71 0. 81 0. 82 0. 80 0. 73 0. 63 0. 82
812 热 带 生 物 学 报 2015 年
续附表 Renew Schedule
编号 S31 S32 S33 S34 S35 S36 S37 S38 S39 S40 S41 S42 S43 S44 S45
S31 1. 00
S32 0. 80 1. 00
S33 0. 78 0. 84 1. 00
S34 0. 76 0. 83 0. 92 1. 00
S35 0. 78 0. 82 0. 94 0. 92 1. 00
S36 0. 78 0. 85 0. 87 0. 90 0. 86 1. 00
S37 0. 78 0. 81 0. 83 0. 83 0. 84 0. 88 1. 00
S38 0. 71 0. 73 0. 75 0. 76 0. 74 0. 78 0. 71 1. 00
S39 0. 78 0. 83 0. 82 0. 83 0. 84 0. 91 0. 85 0. 82 1. 00
S40 0. 78 0. 84 0. 83 0. 88 0. 87 0. 92 0. 86 0. 82 0. 94 1. 00
S41 0. 81 0. 84 0. 78 0. 81 0. 78 0. 83 0. 81 0. 79 0. 83 0. 86 1. 00
S42 0. 81 0. 85 0. 79 0. 79 0. 81 0. 84 0. 83 0. 81 0. 88 0. 86 0. 88 1. 00
S43 0. 80 0. 82 0. 83 0. 84 0. 85 0. 83 0. 87 0. 77 0. 84 0. 87 0. 84 0. 87 1. 00
S44 0. 79 0. 78 0. 81 0. 81 0. 80 0. 76 0. 72 0. 75 0. 79 0. 81 0. 79 0. 79 0. 78 1. 00
S45 0. 72 0. 84 0. 82 0. 83 0. 81 0. 77 0. 75 0. 74 0. 78 0. 81 0. 79 0. 82 0. 81 0. 85 1. 00
S46 0. 74 0. 86 0. 80 0. 83 0. 79 0. 79 0. 80 0. 70 0. 78 0. 81 0. 80 0. 81 0. 79 0. 80 0. 91
S47 0. 74 0. 74 0. 72 0. 73 0. 74 0. 69 0. 66 0. 80 0. 72 0. 74 0. 83 0. 73 0. 71 0. 76 0. 73
S48 0. 72 0. 80 0. 81 0. 79 0. 80 0. 83 0. 79 0. 74 0. 82 0. 81 0. 76 0. 78 0. 81 0. 71 0. 82
S49 0. 74 0. 76 0. 80 0. 80 0. 81 0. 79 0. 80 0. 73 0. 81 0. 81 0. 78 0. 77 0. 81 0. 72 0. 77
S50 0. 68 0. 73 0. 78 0. 75 0. 76 0. 80 0. 78 0. 67 0. 75 0. 75 0. 72 0. 75 0. 78 0. 68 0. 75
S51 0. 81 0. 85 0. 83 0. 80 0. 81 0. 79 0. 81 0. 72 0. 76 0. 78 0. 81 0. 80 0. 81 0. 73 0. 81
S52 0. 81 0. 80 0. 82 0. 81 0. 83 0. 81 0. 82 0. 74 0. 79 0. 82 0. 81 0. 79 0. 81 0. 76 0. 78
S53 0. 76 0. 75 0. 77 0. 77 0. 76 0. 76 0. 77 0. 72 0. 74 0. 77 0. 76 0. 76 0. 78 0. 74 0. 74
S54 0. 76 0. 74 0. 76 0. 75 0. 76 0. 74 0. 76 0. 69 0. 74 0. 74 0. 74 0. 76 0. 76 0. 71 0. 72
S55 0. 80 0. 74 0. 77 0. 74 0. 78 0. 74 0. 73 0. 70 0. 74 0. 74 0. 73 0. 77 0. 75 0. 74 0. 72
S56 0. 73 0. 74 0. 77 0. 74 0. 76 0. 78 0. 78 0. 65 0. 74 0. 74 0. 73 0. 74 0. 75 0. 70 0. 73
S57 0. 74 0. 72 0. 70 0. 70 0. 72 0. 67 0. 73 0. 73 0. 69 0. 70 0. 74 0. 74 0. 74 0. 69 0. 67
S58 0. 79 0. 80 0. 82 0. 78 0. 84 0. 77 0. 79 0. 68 0. 78 0. 78 0. 75 0. 76 0. 78 0. 74 0. 74
S59 0. 76 0. 78 0. 74 0. 73 0. 75 0. 75 0. 76 0. 65 0. 74 0. 74 0. 74 0. 76 0. 75 0. 76 0. 73
S60 0. 71 0. 73 0. 74 0. 75 0. 76 0. 69 0. 69 0. 71 0. 72 0. 72 0. 72 0. 74 0. 73 0. 74 0. 72
S61 0. 76 0. 76 0. 78 0. 75 0. 76 0. 77 0. 76 0. 67 0. 74 0. 75 0. 78 0. 78 0. 78 0. 76 0. 76
S62 0. 74 0. 74 0. 82 0. 81 0. 81 0. 77 0. 81 0. 65 0. 74 0. 78 0. 75 0. 76 0. 80 0. 74 0. 78
S63 0. 79 0. 81 0. 83 0. 78 0. 80 0. 81 0. 83 0. 65 0. 79 0. 79 0. 76 0. 78 0. 81 0. 75 0. 76
912第 2 期 罗 灿等:肾茶种质资源遗传多样性的 ISSR分析
续附表 Renew Schedule
编号 S31 S32 S33 S34 S35 S36 S37 S38 S39 S40 S41 S42 S43 S44 S45
S64 0. 78 0. 81 0. 81 0. 80 0. 82 0. 79 0. 76 0. 76 0. 78 0. 67 0. 80 0. 83 0. 79 0. 76 0. 76
S65 0. 81 0. 81 0. 81 0. 77 0. 82 0. 81 0. 81 0. 67 0. 83 0. 80 0. 78 0. 80 0. 79 0. 77 0. 77
S66 0. 76 0. 77 0. 82 0. 78 0. 78 0. 84 0. 79 0. 69 0. 81 0. 79 0. 76 0. 76 0. 74 0. 75 0. 72
S67 0. 55 0. 54 0. 59 0. 62 0. 61 0. 56 0. 58 0. 58 0. 53 0. 56 0. 52 0. 51 0. 56 0. 55 0. 58
S68 0. 59 0. 56 0. 58 0. 59 0. 60 0. 56 0. 58 0. 58 0. 55 0. 58 0. 56 0. 53 0. 56 0. 58 0. 55
S69 0. 57 0. 56 0. 56 0. 60 0. 58 0. 59 0. 60 0. 57 0. 57 0. 60 0. 57 0. 56 0. 53 0. 57 0. 56
S70 0. 56 0. 56 0. 55 0. 58 0. 57 0. 57 0. 59 0. 56 0. 55 0. 58 0. 55 0. 56 0. 54 0. 56 0. 56
S71 0. 59 0. 57 0. 56 0. 60 0. 60 0. 57 0. 58 0. 56 0. 55 0. 58 0. 58 0. 55 0. 54 0. 58 0. 56
S72 0. 57 0. 56 0. 61 0. 63 0. 63 0. 56 0. 58 0. 58 0. 56 0. 58 0. 56 0. 53 0. 59 0. 57 0. 56
S73 0. 78 0. 75 0. 76 0. 77 0. 78 0. 79 0. 76 0. 76 0. 80 0. 81 0. 80 0. 77 0. 74 0. 74 0. 72
S74 0. 77 0. 75 0. 80 0. 78 0. 81 0. 82 0. 77 0. 76 0. 83 0. 81 0. 80 0. 80 0. 76 0. 73 0. 73
S75 0. 84 0. 72 0. 74 0. 77 0. 75 0. 78 0. 76 0. 69 0. 78 0. 77 0. 78 0. 78 0. 75 0. 76 0. 70
S76 0. 85 0. 74 0. 79 0. 78 0. 80 0. 76 0. 74 0. 76 0. 79 0. 79 0. 79 0. 78 0. 80 0. 81 0. 74
S77 0. 81 0. 72 0. 74 0. 74 0. 74 0. 72 0. 71 0. 68 0. 76 0. 76 0. 76 0. 75 0. 76 0. 85 0. 78
S78 0. 79 0. 73 0. 76 0. 72 0. 76 0. 74 0. 74 0. 75 0. 78 0. 76 0. 78 0. 76 0. 76 0. 75 0. 75
S79 0. 75 0. 77 0. 78 0. 75 0. 78 0. 77 0. 75 0. 74 0. 79 0. 78 0. 75 0. 78 0. 74 0. 74 0. 76
S80 0. 76 0. 78 0. 78 0. 76 0. 79 0. 79 0. 78 0. 69 0. 80 0. 78 0. 74 0. 78 0. 76 0. 72 0. 74
S81 0. 81 0. 74 0. 76 0. 75 0. 78 0. 76 0. 74 0. 69 0. 78 0. 75 0. 72 0. 72 0. 74 0. 76 0. 75
S82 0. 76 0. 78 0. 76 0. 72 0. 77 0. 74 0. 71 0. 75 0. 78 0. 78 0. 75 0. 76 0. 73 0. 78 0. 76
S83 0. 78 0. 83 0. 82 0. 78 0. 81 0. 81 0. 78 0. 71 0. 82 0. 82 0. 75 0. 78 0. 76 0. 72 0. 78
S84 0. 83 0. 85 0. 84 0. 78 0. 83 0. 85 0. 81 0. 67 0. 84 0. 81 0. 77 0. 80 0. 78 0. 76 0. 76
S85 0. 79 0. 80 0. 79 0. 76 0. 78 0. 83 0. 78 0. 71 0. 85 0. 83 0. 81 0. 79 0. 77 0. 76 0. 78
S86 0. 80 0. 82 0. 83 0. 84 0. 83 0. 86 0. 81 0. 72 0. 84 0. 84 0. 83 0. 81 0. 83 0. 76 0. 80
S87 0. 82 0. 83 0. 81 0. 85 0. 81 0. 85 0. 83 0. 68 0. 81 0. 85 0. 82 0. 79 0. 83 0. 79 0. 78
编号 S46 S47 S48 S49 S50 S51 S52 S53 S54 S55 S56 S57 S58 S59 S60 S61
S46 1. 00
S47 0. 74 1. 00
S48 0. 80 0. 67 1. 00
S49 0. 74 0. 71 0. 83 1. 00
S50 0. 72 0. 58 0. 82 0. 83 1. 00
S51 0. 81 0. 71 0. 80 0. 81 0. 76 1. 00
S52 0. 76 0. 73 0. 78 0. 81 0. 79 0. 85 1. 00
022 热 带 生 物 学 报 2015 年
续附表 Renew Schedule
编号 S46 S47 S48 S49 S50 S51 S52 S53 S54 S55 S56 S57 S58 S59 S60 S61
S53 0. 72 0. 72 0. 76 0. 81 0. 76 0. 82 0. 90 1. 00
S54 0. 70 0. 67 0. 75 0. 84 0. 76 0. 83 0. 88 0. 91 1. 00
S55 0. 68 0. 68 0. 76 0. 83 0. 76 0. 81 0. 83 0. 86 0. 90 1. 00
S56 0. 69 0. 60 0. 76 0. 76 0. 84 0. 76 0. 84 0. 78 0. 83 0. 76 1. 00
S57 0. 68 0. 74 0. 66 0. 78 0. 65 0. 78 0. 80 0. 83 0. 84 0. 83 0. 69 1. 00
S58 0. 73 0. 69 0. 78 0. 83 0. 78 0. 83 0. 90 0. 84 0. 85 0. 87 0. 83 0. 81 1. 00
S59 0. 75 0. 64 0. 73 0. 78 0. 80 0. 76 0. 84 0. 82 0. 83 0. 79 0. 82 0. 75 0. 84 1. 00
S60 0. 72 0. 72 0. 72 0. 76 0. 65 0. 76 0. 75 0. 78 0. 78 0. 80 0. 65 0. 83 0. 78 0. 67 1. 00
S61 0. 74 0. 66 0. 72 0. 73 0. 82 0. 78 0. 82 0. 77 0. 79 0. 76 0. 83 0. 67 0. 78 0. 83 0. 67 1. 00
S62 0. 76 0. 63 0. 81 0. 81 0. 82 0. 81 0. 86 0. 84 0. 82 0. 80 0. 83 0. 74 0. 85 0. 84 0. 72 0. 79
S63 0. 76 0. 67 0. 83 0. 83 0. 82 0. 84 0. 88 0. 83 0. 83 0. 84 0. 84 0. 74 0. 88 0. 81 0. 74 0. 83
S64 0. 78 0. 67 0. 80 0. 83 0. 80 0. 81 0. 87 0. 85 0. 87 0. 82 0. 88 0. 74 0. 88 0. 88 0. 72 0. 83
S65 0. 76 0. 69 0. 83 0. 86 0. 81 0. 79 0. 84 0. 81 0. 83 0. 86 0. 82 0. 75 0. 88 0. 83 0. 73 0. 80
S66 0. 72 0. 65 0. 78 0. 78 0. 86 0. 77 0. 81 0. 80 0. 79 0. 77 0. 83 0. 69 0. 79 0. 78 0. 67 0. 83
S67 0. 56 0. 58 0. 59 0. 61 0. 60 0. 60 0. 56 0. 56 0. 59 0. 60 0. 57 0. 60 0. 59 0. 54 0. 56 0. 53
S68 0. 56 0. 63 0. 55 0. 61 0. 56 0. 57 0. 58 0. 57 0. 60 0. 60 0. 54 0. 63 0. 58 0. 56 0. 56 0. 55
S69 0. 56 0. 59 0. 56 0. 59 0. 58 0. 59 0. 56 0. 56 0. 60 0. 59 0. 58 0. 56 0. 57 0. 56 0. 54 0. 58
S70 0. 56 0. 60 0. 55 0. 61 0. 59 0. 60 0. 56 0. 57 0. 60 0. 61 0. 56 0. 58 0. 58 0. 54 0. 56 0. 59
S71 0. 57 0. 63 0. 55 0. 61 0. 56 0. 57 0. 56 0. 56 0. 60 0. 58 0. 56 0. 63 0. 60 0. 57 0. 56 0. 53
S72 0. 56 0. 65 0. 56 0. 62 0. 56 0. 59 0. 60 0. 59 0. 61 0. 62 0. 52 0. 66 0. 61 0. 52 0. 58 0. 53
S73 0. 72 0. 71 0. 77 0. 79 0. 72 0. 83 0. 80 0. 79 0. 78 0. 82 0. 72 0. 75 0. 81 0. 76 0. 77 0. 76
S74 0. 69 0. 67 0. 78 0. 79 0. 77 0. 82 0. 81 0. 79 0. 78 0. 81 0. 75 0. 72 0. 81 0. 75 0. 77 0. 74
S75 0. 74 0. 72 0. 74 0. 79 0. 72 0. 76 0. 78 0. 81 0. 80 0. 85 0. 71 0. 74 0. 80 0. 79 0. 72 0. 77
S76 0. 72 0. 76 0. 76 0. 78 0. 68 0. 83 0. 85 0. 84 0. 83 0. 87 0. 73 0. 78 0. 83 0. 74 0. 82 0. 74
S77 0. 77 0. 73 0. 72 0. 74 0. 68 0. 73 0. 75 0. 77 0. 76 0. 77 0. 69 0. 72 0. 75 0. 77 0. 75 0. 75
S78 0. 73 0. 70 0. 81 0. 78 0. 75 0. 80 0. 86 0. 84 0. 85 0. 84 0. 78 0. 77 0. 85 0. 81 0. 75 0. 79
S79 0. 74 0. 70 0. 81 0. 80 0. 71 0. 80 0. 82 0. 84 0. 83 0. 81 0. 76 0. 77 0. 85 0. 80 0. 81 0. 76
S80 0. 71 0. 63 0. 81 0. 79 0. 80 0. 81 0. 85 0. 86 0. 85 0. 83 0. 85 0. 75 0. 88 0. 83 0. 74 0. 80
S81 0. 72 0. 72 0. 78 0. 80 0. 74 0. 74 0. 81 0. 83 0. 81 0. 84 0. 77 0. 74 0. 83 0. 80 0. 75 0. 76
S82 0. 73 0. 74 0. 75 0. 78 0. 67 0. 80 0. 83 0. 84 0. 82 0. 81 0. 76 0. 77 0. 83 0. 78 0. 76 0. 71
S83 0. 74 0. 67 0. 79 0. 80 0. 76 0. 78 0. 82 0. 80 0. 79 0. 81 0. 81 0. 73 0. 86 0. 77 0. 74 0. 78
S84 0. 75 0. 68 0. 80 0. 79 0. 74 0. 79 0. 80 0. 78 0. 77 0. 79 0. 76 0. 71 0. 83 0. 79 0. 73 0. 78
S85 0. 76 0. 70 0. 82 0. 81 0. 75 0. 78 0. 83 0. 84 0. 82 0. 83 0. 78 0. 76 0. 83 0. 81 0. 74 0. 81
S86 0. 76 0. 68 0. 85 0. 83 0. 81 0. 82 0. 83 0. 79 0. 78 0. 81 0. 79 0. 71 0. 84 0. 78 0. 73 0. 80
S87 0. 78 0. 72 0. 79 0. 80 0. 78 0. 80 0. 81 0. 80 0. 78 0. 78 0. 77 0. 70 0. 81 0. 78 0. 74 0. 81
122第 2 期 罗 灿等:肾茶种质资源遗传多样性的 ISSR分析
续附表 Renew Schedule
编号 S62 S63 S64 S65 S66 S67 S68 S69 S70 S71 S72 S73 S74 S75 S76
S62 1. 00
S63 0. 85 1. 00
S64 0. 85 0. 91 1. 00
S65 0. 83 0. 94 0. 92 1. 00
S66 0. 76 0. 86 0. 85 0. 87 1. 00
S67 0. 56 0. 58 0. 56 0. 60 0. 60 1. 00
S68 0. 56 0. 58 0. 57 0. 61 0. 62 0. 94 1. 00
S69 0. 56 0. 58 0. 59 0. 62 0. 65 0. 94 0. 92 1. 00
S70 0. 55 0. 60 0. 57 0. 63 0. 63 0. 93 0. 92 0. 98 1. 00
S71 0. 56 0. 58 0. 60 0. 63 0. 62 0. 93 0. 93 0. 92 0. 92 1. 00
S72 0. 56 0. 58 0. 56 0. 60 0. 60 0. 91 0. 92 0. 88 0. 88 0. 90 1. 00
S73 0. 78 0. 78 0. 81 0. 81 0. 78 0. 60 0. 61 0. 63 0. 63 0. 60 0. 59 1. 00
S74 0. 78 0. 77 0. 78 0. 78 0. 78 0. 57 0. 56 0. 59 0. 58 0. 56 0. 55 0. 90 1. 00
S75 0. 77 0. 80 0. 81 0. 82 0. 78 0. 58 0. 61 0. 62 0. 61 0. 61 0. 60 0. 81 0. 79 1. 00
S76 0. 78 0. 79 0. 77 0. 78 0. 74 0. 53 0. 55 0. 53 0. 53 0. 53 0. 57 0. 87 0. 85 0. 84 1. 00
S77 0. 79 0. 75 0. 77 0. 77 0. 72 0. 52 0. 55 0. 54 0. 53 0. 56 0. 54 0. 78 0. 76 0. 81 0. 88
S78 0. 83 0. 83 0. 83 0. 85 0. 78 0. 56 0. 58 0. 58 0. 56 0. 55 0. 57 0. 81 0. 84 0. 81 0. 85
S79 0. 79 0. 82 0. 83 0. 84 0. 75 0. 55 0. 53 0. 57 0. 56 0. 55 0. 57 0. 81 0. 83 0. 78 0. 82
S80 0. 83 0. 87 0. 88 0. 86 0. 83 0. 57 0. 54 0. 58 0. 56 0. 56 0. 53 0. 78 0. 82 0. 79 0. 78
S81 0. 76 0. 81 0. 83 0. 84 0. 78 0. 59 0. 58 0. 58 0. 58 0. 59 0. 57 0. 74 0. 76 0. 84 0. 83
S82 0. 78 0. 78 0. 80 0. 78 0. 72 0. 52 0. 53 0. 54 0. 53 0. 52 0. 57 0. 80 0. 80 0. 78 0. 83
S83 0. 79 0. 85 0. 84 0. 87 0. 81 0. 59 0. 58 0. 63 0. 60 0. 58 0. 60 0. 76 0. 78 0. 76 0. 75
S84 0. 78 0. 84 0. 85 0. 85 0. 80 0. 51 0. 53 0. 53 0. 53 0. 54 0. 52 0. 79 0. 79 0. 81 0. 81
S85 0. 81 0. 86 0. 87 0. 88 0. 83 0. 58 0. 60 0. 61 0. 59 0. 60 0. 58 0. 81 0. 80 0. 84 0. 81
S86 0. 84 0. 84 0. 83 0. 86 0. 83 0. 60 0. 57 0. 60 0. 58 0. 60 0. 58 0. 81 0. 82 0. 83 0. 80
S87 0. 82 0. 85 0. 84 0. 84 0. 82 0. 60 0. 60 0. 64 0. 62 0. 62 0. 58 0. 80 0. 77 0. 84 0. 79
编号 S77 S78 S79 S80 S81 S82 S83 S84 S85 S86 S87
S77 1. 00
S78 0. 82 1. 00
S79 0. 79 0. 92 1. 00
S80 0. 72 0. 87 0. 88 1. 00
S81 0. 81 0. 85 0. 85 0. 87 1. 00
S82 0. 76 0. 83 0. 86 0. 83 0. 81 1. 00
S83 0. 72 0. 86 0. 89 0. 90 0. 86 0. 82 1. 00
S84 0. 80 0. 77 0. 78 0. 83 0. 83 0. 78 0. 84 1. 00
S85 0. 79 0. 85 0. 83 0. 88 0. 85 0. 83 0. 88 0. 90 1. 00
S86 0. 77 0. 83 0. 83 0. 88 0. 81 0. 78 0. 85 0. 88 0. 88 1. 00
S87 0. 81 0. 79 0. 81 0. 83 0. 81 0. 76 0. 83 0. 84 0. 85 0. 94 1. 00
222 热 带 生 物 学 报 2015 年