我国石斛属植物种质资源及其亲缘关系研究
详细信息    本馆镜像全文|  推荐本文 |  |   获取CNKI官网全文
摘要
石斛属(Dendrobium)植物是兰科的附生植物,它是我国传统的名贵中药材,除了具有较高的药用价值外,它还具有较高的观赏价值。现在广泛栽培的石斛兰是以石斛属植物的原生种为亲本进行杂交选育出的优良观花品种。我国有丰富的石斛属植物资源,但观赏石斛的育种比较落后。针对这一问题,本研究进行了我国石斛属植物的种质资源及其亲缘关系的研究,以期为我国的观赏石斛兰育种奠定基础。
     本研究调查研究了我国石斛属植物资源现状及其保护措施,并从思茅、景洪和深圳等地收集引种石斛属植物31种1变种,并对其观赏特性进行综合评价。采用石斛叶绿体DNA trnL-trnF间隔区序列比较和采用ISSR分子标记对我国的部分石斛属植物资源进行亲缘关系分析,主要研究结果如下:
     调查收集并引种栽培了31个石斛原生种和1个变种,引种的石斛资源栽培于中国林业科学研究院科研温室。
     首次运用模糊数学的方法对所收集的石斛资源进行观赏特性综合评价。将32个石斛样本依据观赏特性划分为优、良、中、差4个等级。综合观赏特性优的有7种石斛;良的有14种石斛和1个变种;中等的有8种石斛;差的有2种石斛。
     对利用CTAB法从以下几种石斛(铁皮石斛、石斛、细茎石斛、齿瓣石斛、喉红石斛、黑毛石斛、翅梗石斛)的幼嫩叶片提取DNA的方法进行了改进,用改进后的方法提取的DNA数量多,质量好,纯度高。
     首次利用石斛属植物的叶绿体DNA trnL-trnF间隔区序列分析研究石斛属植物种间的亲缘关系。所测的30个石斛样本的叶绿体DNA trnL-trnF间隔区序列已登录GenBank。
     所测序列用CLUSTAL软件进行序列分析,分析了石斛种间的亲缘关系,总体上将30个石斛样本划分为4个相对独立的大组。
     石斛叶绿体DNA trnL-trnF间隔区序列分析确定出的石斛属植物部分种间的亲缘关系为:报春石斛和兜唇石斛亲缘关系较近;串珠石斛与大苞鞘石斛亲缘关系较近;黑毛石斛与长距石斛和喉红石斛亲缘关系较近;刀叶石斛与剑叶石斛亲缘关系较近;这与中国植物志石斛属形态分类相一致。而翅梗石斛与小黄花石斛亲缘关系较近;鼓槌石斛与短棒石斛亲缘关系较近;石斛与重唇石斛亲缘关系较近这与中国植物志的石斛属形态分类不一致。
     首次从60条ISSR引物中筛选出5条引物用于石斛属植物ISSR-PCR扩增。这5条引物扩增的条带清晰、多态性好,32个石斛样本共扩增出103个清晰、稳定的条带,其中有93个多态性条带,多态性条带百分率为90.2%,扩增的条带长度在200—2000bp之间。建立了稳定的、可重复的石斛属植物ISSR-PCR反应体系,为今后利用ISSR标记技术开展石斛种间亲缘关系的研究提供一个标准化程序。
     根据ISSR扩增结果,用Ntsys(2.0)软件计算出遗传相似系数和遗传距离,按UPGMA方法构建聚类树状图,32个石斛样本可分为4组。
     利用ISSR分子标记确定出的石斛属植物部分种间的亲缘关系为:细茎石斛与铁皮石斛和滇桂石斛亲缘关系较近;流苏石斛与叠鞘石斛亲缘关系较近;喉红石斛与黑毛石斛和长距石斛亲缘关系较近;刀叶石斛与剑叶石斛亲缘关系较近。这与中国植物志石斛属形态分类相一致。而翅梗石斛与小黄花石斛亲缘关系较近;束花石斛与罗河石斛亲缘关系较近;这与中国植物志石斛属形态分类不一致。
     两种分子标记方法都支持的石斛属植物亲缘关系为:黑毛石斛与长距石斛和喉红石斛亲缘关系较近;刀叶石斛与剑叶石斛亲缘关系较近;这与中国植物志石斛属形态分类相一致。翅梗石斛与小黄花石斛亲缘关系较近;鼓槌石斛与短棒石斛亲缘关系较近;这与中国植物志石斛属形态分类不一致。
Dendrobium species are epiphytic plants of orchidaceae. Many wild species ofDendrobium are traditional precious Chinese medical herbals and precious ornamentalplants.Dendrobium orchid cultivers which are cultivated diffusely are artifical hybrids ofDendrobium species.There are rich Dendrobium resources in China, but Densrobium orchidbreeding in China drop behind other countries. Aim at this problem, this research studies on thegermplasm resources of Dendrobiun in China and their genetic relationships, in order toestablish the base of Dendrobium orchid breeding in China.
     The status quo of Dendrobium resources in China and their conservation measures wereinvestigated and studied. 31 Dendrobium species and 1 variety were collected from Simao andJinghong, Yunnan province and Shenzhen, Guangdong province. The ornamentalcharacteristics of the 32 spcimen were evaluated. Marking by the chloroplast DNA (cpDNA)trnL-trnF sequence and the inter-simple sequence repeat (ISSR) markers, their geneticrelationships were conducted. The main results were as follows.
     31 Dendrobium species and 1 variety were introduced and cultivated in the greenhouse forscience research in Chinese academy of forestry.
     It is the first time to evaluate the ornamental characteristics of the Dendrobium resourcesin China by fuzzy mathematics. 32 Dendrobium spp. samples were classified into 4 groups bytheir ornamental characteristics. GroupⅠ,7 species, which are best; GroupⅡ, 14 species and 1variety, which are better; GroupⅢ,8 species, which are medium; GroupⅣ,2 species, whichare disqualification.
     Ameliorated the DNA extraction method by CTAB from the young foliages of someDendrobium species, such as D. officinale, D. nobile, D. moniliforme, D. devonianum, D.christyanum, D. williamsonii, D. trigonopus.
     And it is also the first time to study on the genetic relationships inter the Dendrobiumspecies in China by chloroplast DNA (cpDNA) trnL-trnF sequence. 30 cpDNA trnL-trnFsequences of the Dendrobium resource samples had been accepted by GenBank of NCBI.
     The genetic relationships of 30 samples were analyzed by using software CLUSTALbased on the chloroplast DNA (cpDNA) trnL-trnF sequence.
     The results showed that 30 samples were classified into 4 groups. The geneticrelationships of 30 Dendrobium samples are as follow. D. primulinum is close with D. aphyllum; D..falconeri is close with D. wardianum; D. williamsonii is close with D. longicornuand D. christyanum; D. terminale is close with D. acinaciforme. These are according to withthe morphological taxonomy of Dendrobium of Chinese flora sinica. But D. trigonopus is closewith D. jenkinsi; D. chrysotoxum is close with D. capillipes; D. nobile is close with D.hercoglossum. The results are not support the Chinese flora sinica classification which isclassified the Dendrobium with the morphological taxonomy.
     The genetic relationships of Dendrobium resource in China were studied by ISSR marker.5 primers, which produced clear and rich polymorphism bands were chosen from 60ISSR-PCR primers. 103 bands were amplified from 32 samples, and 93 bands of them werepolymorphic (percentage of polymorphic bands was 90.2%). The length of the amplified bandswere between 200~2000bp. The stable and reproducible optimum reaction system ofISSR-PCR and PCR amplification parameter were established. The results provided astandardizing program for the analysis of inter species genetic relationships of Dendrobiumresources.
     According to the results of ISSR amplified, similarity coefficient and genetic distancewere calculated by using Ntsys(2.0) software.A dendrogram was constructed based onUPGMA cluster analysis. 32 Dendrobium spp. samples were classified into 4 major groups.The genetic relationships of Dendrobium species in China were as fallow: D. moniliforme isclose with D. officinale and D. guangxiense; D. fimbriatum is close with D. denneanum; D.christyanum is close with D. williamsonii and D. longicornu; D. terminale is close with D.acinaciforme. The result is coherency with the Chinese flora sinica. But D. trigonopus is closewith D. jenkinsii; D. chrysanthum is close with D. lohohense. It is incoherecy with theChinese flora sinica.
     Based on the results of two molecular markers, the genetic relationships of Dendrobiumspecies in China were as follow: D. williamsonii is close with D. longicornu and D.christyanum; D. terminale is close with D. acinaciforme. It is coherency with the Chinese florasinica. But D. trigonopus is close with D. jenkinsii; D. chrysotoxum is close with D. capillipes.It is incoherecy with the Chinese flora sinica.
引文
[1] 吉占和,陈心启,罗毅波,等.中国植物志.第19卷.北京:科学出版社,1999.67~257
    [2] 高江云.西双版纳石斛资源的保护与利用.园艺学报,1996,23(2):160
    [3] 陈心启,吉占和,罗毅波.中国野生兰科植物彩色图鉴.北京:科学出版社,1999
    [4] 曾宋君,胡松华.石斛兰.广州:广东科技出版社,2004
    [5] 包雪声,顺庆生,陈立钻.中国药用石斛彩色图谱.上海:上海医科大学出版社,复旦大学出版社,2001.1~108
    [6] 陈心启,吉占和.中国兰花全书.北京:中国林业出版社,1998.153~171
    [7] 卢思聪.中国兰与洋兰.北京:金盾出版社,1994
    [8] 吉占和.中国石斛属的初步研究.植物分类学报,1980,18(4):427~449
    [9] Bill Lavarack,Wayne Harris,Geoff Stocker.DENDROBIUM AND ITS RELATIVES.Portland, Oregon, U,S,A:Timber Press,2002
    [10] Kraenzlin,F.Orchidaceae-Monandrae-Dendrobiina I in A.Engler, Das Pflanzenreich Heft, 1910,45
    [11] Smith.J.J. Die Orchidaceae Von Java, 1905,306~374
    [12] Schlechter R. Die Orchidaceae Von Deutsch-Neu-Guinea Feddes Repert Sp.Nov. Beih. 1914,1:569~642
    [13] Holttum R.E Flora of Malaya Orchida of Malaya,1953 1:258~339
    [14] 刘晓燕.泰国兰花生产考察与思考.贵州农业科学,2005,33(2):100~102
    [15] Goh C J, Yang A L. Effects of growth regulators and decapitation on flowering of Dendrobium orchid hybrids.Plant-Science-Letters, 1978,12(3):287~292
    [16] Roedjito S.W. Artificial media for Dendrobium orchid nursery. Berita-Biologi, 1979,2(4) :84
    [17] Porat,R,Borochov A,Halevy A H.Pollination-induced changes in ethylene production and sensitivity to ethylenein cut dendrobium orchid flowers.Scientia-Horticulturae, 1994,58(3) :215~221
    [18] Widiastoety D, Bahar F A. Effect of different sources and dosages of carbohydrate on the growth of plantlet of Dendrobium orchid. Jurnal-Hortikultura ,1995,5(3) :76~80
    [19] Mamaril J.C., Lopez A.M. Effect of growth hormone extracts from coconut water on the growth of reflasked dendrobium protocorm .Philippine Journal of Crop Science, 1996, 21 (Supplement no. 1 ): 30
    [20] Vellupillai M, Swarup S, Chong-JinGoh. Histological and protein changes during earlystages of seed germination in the orchid, Dendrobium crumenatum.Journal of Horticultural Science,1997,72(6):941~948.
    [21] Khoo, G.H., Hew C.S,He J. Photosynthetic utilization of radiant energy by CAM Dendrobium flowers. Photosynthetica, 1997, 34(3) :367~376.
    [22] Widiastoety D, Kusumo S, Syafni .The influence of water coconut maturity and coconut variety on the growth of Dendrobium orchid plantlets. Jurnal Hortikultura 1997,7(3) :768~772
    [23] Mitra A, Dey S, Sawarkar S K. Photoautotrophic in vitro multiplication of the orchid Dendrobium under CO2 enrichment. Biologia Plantarum, 1998,41 (1): 145~148
    [24] Widiastoety D., Prasetio W, Solvia N.The influence of shading on the flowers production of three cultivars of Dendrobium orchid.JurnaI-Hortikultura,2000,9(4): 302~306
    [25] Prasad G.V.S. Sai, Rao I.V.Subba, Reddy P Veera.ln vitro propagation of orchid-Dendrobium 'Sonia'. Indian Journal of Plant Physiology, 2001,6(3): 284~288
    [26] Porat R, Borochov A, Halevy A.H. is jasmonic acid involved in the endogenous regulation of Dendrobium orchid flower senescence. Acta-hortie. Leuven, Belgium: International Society for Horticultural Science, 1995, (405): 314~319.
    [27] Ketsa S, Thampitakorn F. Characteristics of ethylene production of Dendrobium orchid flowers. Actaohortic. Leuven, Belgium : International Society for Horticultural Science. Oct 1995. (405): 253~263
    [28] Kuehnle A R,Lewis D.H.,Markham K R,Mitchell K A,Davies M,Jordan B.R. Floral flavonoids and pH in Dendrobium orchid species and hybrids. Euphytica. Dordrecht, 1997,95 (2) :187~194.
    [29] Saiprasad G.V.S., Polisetty Raghuveer, Raj Anupam. Effect of growth regulators on production of PLBs and multiple shoots in orchid: Dendrobium 'Sonia': Assessment of role of methane and ethylene. Phytomorphology,2003,53(1): 63~71
    [30] Saiprasad G.V.S., Raghuveer P, Khetarpa S ,Chandra R. Effect of various polyamines on production of protocorm-like bodies in orchid: Dendrobium 'Sonia'. Scientia-Horticulturae,2004,100(1-4): 161~168
    [31] Haruyuki Kamemoto,Teresita D. Amore,and Adelheid R. Kuehnle,Breeding Dendrobium Orchids in Hawaii.Honolulu:University of Hawaii Press, 1999
    [32] 唐振缁,程式君.广东石斜属(兰科)的研究.中国科学院华南植物研究所集刊,1986年
    [33] 卢炯林,高立献.河南石斛属植物的调查研究.武汉植物学研究,1991,9(2):148~152
    [34] 倪勤武,来平凡.浙江富阳发现野生铁皮石斛新分布.业科学研究,2000,3(2):222
    [35] 彭锐,范俊安,张艳.斛属药用植物种质资源研究进展.时珍国医国药,2001,12(3):273~274
    [36] 李秀.云南思茅野生兰科石斛属植物.思茅师范高等专科学校学报,2002,18(3):76~79
    [37] 杨琪.浅议云南兰科植物种质资源保存和种源基地建设.林业调查规划,2002,27(增刊):16~19
    [38] 张铁.文山石斛属药用资源调查与开发利用.保山师专学报,2003,22(5):01~05
    [39] 管艳红,马洁,张丽霞,等.西双版纳石斛属药用植物资源.中国野生植物资源.2004,23(6):38~39
    [40] 姚能昌.浅述云南石斛资源现状及开发利用技术.林业调查规划,2004,29(4):80~82
    [41] 徐兴,李黎.云南石斛的保护和可持续利用.森林与人类.2004,5:52~53
    [42] 邓朝仪,聂建平,卢永成等.贵州石斛属植物资源及其开发利用价值评价.贵州林业科技,2004,32(1):51~53
    [43] 淳泽.药用石斛的资源危机与保护对策.资源开发与市场.2005,21(2):139~140
    [44] 邓君浪,郑宽瑜,赵辉,等.云南药用石斛产业化存在的问题及对策.云南农业科技,2005,(4):7~8
    [45] 蒋波,詹源庆,黄捷.金钗石斛濒危原因及其野生资源保护.中国野生植物资源,2005,24(5):34~36
    [46] 袁玉美.潞西市石斛种质资源及栽培技术.林业调查规划,2005,30(2):112~114
    [47] 白音,包英华,金家兴,等.我国药用石斛资源调查研究.中草药,2006,37(9):4~6
    [48] 苏惠,杨云.纳板河自然保护区石斛属植物资源现状与保护对策。林业调查规划,2006,31(5):100~102
    [49] 李满飞,徐国钧,平田义正,等.中药石斛类多糖的含量测定.中草药,1990,21(10):10
    [50] 毕志明,王峥涛,张勉,等.流苏石斛化学成分的研究(Ⅱ).中国药科大学学报,2001,32(6):421~422
    [51] 杨虹,龚燕晴,王峥涛,等.鼓槌石斛化学成分的研究.中草药,2001,32(11):972~974
    [52] 邓银华,徐康平,谭桂山.石斛属植物化学成分与药理活性研究进展.中药材,2002,25(9):677~680
    [53] 黄民权,阮金月.铁皮石斛氨基酸组分分析.中药材,1997,20(1):32~33
    [54] 王康正,高文远.石斛属药用植物研究进展.中草药,1997,28(10):633~
    [55] 徐珞珊,徐国钧,沙文兰,等.中药石斛显微鉴定研究Ⅰ.南京药学院学报,1980,15(2):1
    [56] 徐珞珊,徐国钧,林惠蓉,等.中药石斛显微鉴定研究Ⅱ.南京药学院学报,1981,17(2):52
    [57] 李满飞,徐国钧,徐珞珊,等.商品石斛的调查及鉴定(Ⅱ).中草药,1991,22(4):173
    [58] 刘学平,汤明辉,戴涌,等中药石斛类粉末的显微鉴定研究冲国药科大学学报,1992,
    [59] 马国祥,徐国钧,徐珞珊,等.商品石斛的调查及鉴定(Ⅲ).中草药,1995,26(7):370
    [60] 张智,翟立业.霍山石斛营养器官的解剖结构.安徽农业大学学报,1995,22(3)
    [61] 何静波.黑节草原胚体的繁殖.云南植物研究,1982,4(2):211~212
    [62] 徐云娟.霍山石斛种子的萌发和试管苗的培养.安徽农学院学报,1984,(1):48~52
    [63] 刘瑞驹,蒙爱东,邓锡青,等.铁皮石斛试管苗快速繁殖的研究.药学学报,1988,23(8):636~640
    [64] 顾慧芬,忻晓君,周文婷,等。铁皮石斛试管苗快速生长与栽培研究及多糖含量测定.中成药1999,21(12):6658~659
    [65] 周根余,谢薇,程磊.影响铁皮石斛原球茎生长的若干因素.江西科学,1999,17(4):231~235
    [66] 张铭,朱峰,魏小勇,等.铁皮石斛种胚萌发和原球茎质量控制.浙江大学学报(理学版)2000,27(1):92~94
    [67] 陈薇,寸守铣.铁皮石斛茎段离体快繁.植物生理学通讯,2002,38920:145
    [68] 蒋林,丁平,郑迎冬.添加剂对铁皮石斛组织培养和快速繁殖的影响.中药材,2003,26(8):539~541
    [69] 王碧琴,盖安俊.铁皮石斛的组织培养及快速繁殖.江西科学,2006,24(6):479~484
    [70] 杨联合.曲茎石斛组织培养研究.中国中药杂志,1998,23(11):658
    [71] 曾宋君,程式君,张京丽,等.五种石斛兰的胚培养及其快速繁殖研究.园艺学报,1998,25(1):75~80
    [72] 宋锡全,宋琴曲.金钗石斛茎段培养再生绿色植株.贵州师范大学学报(自然科学版)2003,21(3):80~82
    [73] 孙廷,杨玉珍,胡如善,等.金钗石斛的组织培养和快繁技术.西北农业学报,2004,13(4):13~16
    [74] 黄肇宇,蒋波,杨存亮等.金钗石斛老茎诱导无菌苗的初步研究.玉林师范学院学报(自然科学版)2004,25(5):76~79
    [75] 黎建岭,黄肇宇,詹源庆,等.金钗石斛试管苗生根研究.广西科学院学报,2006,22(2):87~89
    [76] 宋锡全,吴辉,赵俊.金钗石斛试管苗壮苗培养和光合速率检测.贵州师范大学学报(自然科学版),2006,24(1):6~9
    [77] 蒋波,黄肇宇,梁泽华,等.金钗石斛试管苗移栽基质的初步研究.玉林师范学院学报(自然科学)2006,27(3):98~100
    [78] 徐红,刘峻,王峥涛,等.鼓槌石斛组织培养研究.中国中药杂志,2001,26(6):378~381
    [79] 刘勇,刘贤旺.重唇石斛种子试管苗培养简报.江西中医学院学报,2004,16(1):54
    [80] 丁长春.齿瓣石斛的胚培养技术及其快速繁殖研究.云南农业科技,2004,6:20
    [81] 陈之林,曾宋君,段俊.独角石斛的离体繁殖.植物生理学通讯,2004,40(3):342
    [82] 卢文芸,唐金刚,乙引,等.五种药用石斛快速繁殖的研究.种子,2005,24(5):23~25
    [83] 张明,别之龙,陈仕江,等.温度和光强对金钗石斛吸收磷素的影响.西南农业大学学报,1999,21(3):220~222
    [84] 别之龙,张明,陈仕江,等.金钗石斛对磷素的吸收和运转.核农学报,2002,16(3): 162~166
    [85] 张明,陈仕江,万兆良,等.光照强度对金钗石斛吸收氮素的影响.西南农业大学学报,2000,22(2):108~111
    [86] 陈仕江,张明,丑敏霞,等.金钗石斛生长的最适光温研究.中国中药杂志,2002,27(7):509~510
    [87] 别之龙,万兆良,张明,等.金钗石斛光合产物的运转和分配.核农学报,2001,15(1):51~56
    [88] 张明,陈仕江,李泉森,等.金钗石斛驯化栽培的基质研究,中药材,2001,24(9):628
    [89] 张明,陈仕江,李泉生,等.不同栽培条件下金钗石斛总生物碱含量比较.中药材,2001,24(10): 707~708
    [90] 蔡永萍,李玲,林毅,等.霍山三种石斛的内源GA3、ABA含量对茎高生长的影.激光生物学报,2004,13(5):345~348
    [91] 刘晓青,周建涛.外源GA_3对春石斛园艺性状的影响.江苏农业科学,2004,5:77
    [92] 蔡永萍,李合生,骆炳山,等.霍山3种石斛的生长节律及其与生态因子关系的研究.武汉植物学研究,2003,21(4):351~355
    [93] 苏文华,张光飞.铁皮石斛叶片光合作用的碳代谢途径.植物生态学报,2003,27(5):631~637
    [94] 吴显芝,乙引.外源Ca~(2+)对干旱胁迫下金钗石斛幼苗生长的影响.贵州师范大学学报(自然科学版),2006,24(2):23~26
    [95] 郭顺星,曹文芩,高微微,等.铁皮石斛及金钗石斛菌根真菌的分离及其生物活性测定.中国中药杂志,2000,25(6):338~341
    [96] 胡天放.霍山石斛的野生变为栽培的经验介绍.中药通报,1958,4(12)424
    [97] 林深.石斛贴树培育的经验.药学通报,1959,7(4):157~158
    [98] 王用平.贵州省赤水县石斛栽培方法.中草药通讯,1978,9(7):44~45
    [99] 王用平.贵州产石斛属植物引种驯化研究.贵州药讯,1985,(2):13~16
    [100] 程式君.石斛属的引种栽培.植物引种驯化集刊.第四集.北京:科学出版社,1985.59
    [101] 石友亭.曲茎石斛放养研究.中国中药杂志;1995,(3):
    [102] 张炳言,封光伟,何洪中,等.模拟曲茎石斛生境的栽培技术研究.中药材,1999,22(5):217~220
    [103] 付开聪,冯德强,张绍云,等.铁皮石斛集约化高产栽培技术研究.中草药,2003,34(2):177~179
    [104] 邓君浪,郑宽瑜,赵辉.铁皮石斛集约化栽培管理技术.云南农业科技,2005,(1):33~34
    [105] 唐德英.野生鼓槌石斛引种和栽培技术,中国中药杂志,2006,31(15):1297~1298
    [106] 练强,李俊.齿瓣石斛人工授粉试验,热带农业科技2003,26(2):44~45
    [107] 李涵,郑思乡,李枝林,等.齿瓣石斛多倍体育种研究初报.中国农学通报.2004,20(4):198~199
    [108] 唐树梅,漆智平.石斛兰营养特性及施肥技术初探周艺学报,1999,26(3):184~187
    [109] 刘晓青,周建涛,王燕敏.高档盆栽兰花春石斛栽培管理技术.安徽农业科学,2004,32(5):1093~1094
    [110] 李振坚,王雁.春石斛的花期调控.中国花卉园艺,2005,(8):20~23
    [111] 乔佳伟.春石斛栽培要点.中国花卉园艺,2005,8:19~20
    [112] 李振坚.春石斛规模化繁殖要领.中国花卉园艺,2006,(22):20~22
    [113] 王韬,郑平,赵贵林,等.华南地区春石斛的特征特性及栽培管理.广东农业科学2006,8:41~43
    [114] 陈艳,张晓燕.石斛属植物主要害虫及其风险分析.武夷科学,2002,18:8~14
    [115] 曾宋君,刘东明.石斛兰的主要病害及其防治.中药材,2003,26(7):471~474
    [116] 曾宋君,刘东明,段俊.石斛兰的主要虫害及其防治.中药材,2003,26(9):619~621
    [117] 董诗韬.石斛主要病害及其综合防治技术,2005,30(1):76~79
    [118] 周家琪.牡丹,芍药花型分类的探讨,园艺学报,1962,351~360
    [119] 马燕,毛汉书,陈俊愉.部分月季花品种的数量分类研究,西北植物学报,13(3):225~231
    [120] 张忠义,鲁琳,武荣花,等.牡丹品种种质资源评估模型.生物数学学报,1997,12(4):376~379
    [121] 施修杰,张治明.紫薇(Lagerstroemia indica)种质资源的数量研究.植物引种驯化集刊.第11集,1997,81~92
    [122] 封培波.上海地区引种宿根花卉观赏性评价及耐热抗寒研[硕士学位论文].北京:北京林业大学,2003.8
    [123] 魏娜.北京地区引种奥运宿根花卉观赏性评价及抗寒性研[硕士学位论文].北京:北京林业大学,2006.13
    [124] 杨显志,邵华,周成,等.生物技术在药用石斛研究中的应用.中草药,2002,33(2):173~175
    [125] 杨翠芹,刘伟,王声斌,等.转基因技术在兰花上的应用.亚热带植物科学,2004,33(2):64~68
    [126] 魏小勇,方花.石斛属药用植物分子生物学研究.上海中医药大学学报,2005,19(1)47~49
    [127] 宋喜强,罗毅波,钟云芳,等.石斛属植物生物技术研究概况,园艺学报,2005,32(4):741~747
    [128] 郭顺星,曹文岑,张集慧,等.铁皮石斛人工种子制作流程及发芽研究.中草药,1996,27920:105~107
    [129] 张铭,魏小勇,黄华荣.铁皮石斛人工种子固形包埋系统的研究.园艺学报,2001,28(5):435~439
    [130] 王君晖,张毅祥,刘峰,等.铁皮石斛种子、原球茎和类原球茎体的超低温保存研究.园艺学报,1999,26(1):59~61
    [131] 王光远,刘培,许智宏,等.石斛离体培养中ABA对诱导花芽形成的影响.植物学报,1995,37(5): 374~378
    [132] 温云飞,鲁润龙,谢子立,等霍山石斛的快速繁殖和花芽诱导.植物生理学通讯.1999,35(4):296~297
    [133] 陈肖英.霍山石斛试管开花研究.:[硕士学位论文].广州:华南师范大学,2003.5
    [134] 王琳.金钗石斛试管开花研究:[硕士学位论文].广州:华南师范大学,2004.4
    [135] 王光远,许智宏,蔡德发,等.铁皮石斛的离体开花.中国科学(C辑),1997,27(3):229~234
    [136] 丁兰,付庭治.兰花生物工程研究进展.西北师范大学学报(自然科学版),2000,36(3):111~114
    [137] 魏小勇,方花.石斛属药用植物分子生物学研究。上海中医药大学学报,2005,19(1):47~49
    [138] 庄军平,黄胜琴,潘舒群,等.金钗石斛花芽cDNA表达文库的构建及鉴定.园艺学报,2006,33(4): 895~897
    [139] Yang HH,Chua NH.Isolation and characterization of genes involved in the pigment biosynthesis of orchids. Proceedingsof 13th World Orchid Conference,1990,48~50
    [140] Kuehnle AR, Cheng S,Zhang G, et al.Transformation of Dendrobium orchid using particle bombardment of protocorms. Plant Cell Reports,1992,11(9):484488
    [141] Chia T F, Chan Y.S. ,Chua N H..The firefly luciferase gene as a non-invasive reportor for dendrobium transformation. Plant Journal, 1994,6(3):441~446
    [142] Yu Z H, Chen M.Y, N ie L.Recovery of transgenie orchid plants with hygromycin selection by particle bombardment to protocorms. Plant Cell,Tissue and Organ Cuitre, 1999,58:87~92
    [143] Nan G.L., Tang C.S. Kuehnle A R ,Kado C I. Dendrobium orchids contain inducer of Agrobaterium virulence genes. Physiological an d Molecular Plant Pathology,1997,51(6): 391—399
    [144] Yu H,Yang S H,Goh C .J.Agrobacteriurn-mediated transformation of a Dendrobium orchid with the class 1 knox gene DOH1. Plant Cell Reports,2001, 20: 301~305
    [145] Men S Z,Ming X T,Liu R W . Agrobacterium-mediated genetic transformation of a Dendrobium orchid. Plant Cell,Tissue and Organ Culture. 2003,75:63~71
    [146] 杨翠琴,秦耀国,刘伟.超声波处理对根癌农杆菌介导转化金钗石斛影响的初步研究.四川农业大学学报,2006,24(4):455~458
    [147] Nagl,-W. Heterochromatin elimination in the orchid Dendrobium. Protoplasma. 1983,18 (3) :234~237
    [148] Ranganath R.M., Shubhakara G . Ultrastructural studies in the orchid Dendrobium Queen Sonia. Beitraege zur Biologie 2000,72(1): 15~22
    [149] Yu H, Goh C J. Differential gene expression during floral transition in an orchid hybrid Dendrobium Madame Thong-ln .Plant Cell Reports,2000,19(9): 926~931
    [150] Liu Zhi-xin, Wu-Hao,Pan Jun-song, Zheng Xue-qin. Cloning and sequencing of movement protein gene of cymbidium mosaic virus.Virologica Sinica, 2001,16(1): 51~54
    [151] Wu Xue-Min,Lim Saw-Hoon, Yang Wei-Cai.Characterization, expression and phyiogenetic study of R2R3-MYB genes in orchid. Plant Molecular Biology, 2003,51(6): 959-972
    [152] Yang Shu-Hua, Yu Ha, Gob Chong-Jin .Functional characterisation of a cytokinin oxidase gene DSCKX1 in Dendrobium orchid. Plant Molecular Biology, 2003,51(2): 237~248
    [153] Yang Shuhua, Yu Hao ,Xu Yifeng,Goh Chong-Jin.Investigation of cytokinin-deficient phenotypes in Arabidopsis by ectopic expression of orchid DSCKX1. FEBS-Letters, 2003,555(2): 291~296
    [154] Mudalige Jayawickrama, Rasika G ,Champagne Michele M,Hieber A David,Kuehnle Adelheid R.CIoning and characterization of two anthocyanin biosynthetic genes from Dendrobium orchid .journal of the American Society for Horticultural Science,2005,130(4): 611~618
    [155] Williams JG, Kubelik AR, Livak KJ, et al. DNA polymorphsms amplified by arbitrary primers are useful as genetic markers. Necleic Acids Res,1990,18 (22):6531~6535
    [156] Zabeau M,Vos P.Selective restriction fragment amplification: a general method for DNA fingerprinting .European Patent Application Number:92402629.7,Publication NumberEP0534858 A 1,1993
    [157] Zietkiewicz E, Rafalske A, Labuda D. Genome fingerprinting by simple sequence repeat (SSR)-anchored polymerase chain reaction amplification.Genome.1994,20:178~183
    [158] 黄瑶.叶绿体DNA及其在植物系统学研究中的应用.植物学通报,1994,11(2):11~25
    [159] Donald JP, Donald HL, Garrette C.Phylogenetic Relationships in Nuphar (Nymphaea): Evidence from Morphology Chloroplast DNA and Nuclear Ribosomal DNA.Ameriean.Journal of Botany, 1999,86 (9): 1316~1324
    [160] Tao S,Daniel J Crawford .Chloroplast DNA phylogeny ,reticulate evolution ,and biogeography of Paeonia (Paeoniaceae).American Journal of Botany, 1997,84 (9): 1120~1136
    [161] Terry RG, Robert SN,Robin JT, et al.Genetic Variation in Chloroplast and Nuclear Ribosomal DNA in Utah Juniper(Juniperus Osteosperma, Cupressaceae):Evidence for lnterspecific Gene Flow.American Journal of Botany,2000,87(2):250~258
    [162] Rajora OP, Dancik BP, Chloroplast DNA variation in Populus.lnter specific restriction fragment polymorphisms and genetic relationships among Populus deitoids,P.nigra,P.maximowiczii, and P.Canadensis.Theor Appl Genet, 1995,90:324-330
    [163] Taberlet P, Gielly L,Pautou Get aI.Universal primers for amplification of three non-coding regions of chloroplast DNA. Plant Mol., 1991,17: 1105-1109
    [164] Pelser PB,Gravendeel B,Meijden R.Tackling speciose genera:species composition and polygenetic position of Senecio sect.Jacobaea based on plastid and nrDNA sequences.American Journal of Botany,2002,89:929-939
    [165] Bohe,UR, Hilger H,Cerff R, et al. Noncoding chloroplast DNA for plant molecular systematics at the infrageneric level. In Schierwater B,Streit B,Wagner CP, Molecular ecology and evolution: approaches and applications, 1994,391-403
    [166] Gielly, L,Taberlet EThe use of chloroplast DNA to resolve plant phylogenies:noncoding versus rbcl sequences.Molecular Biololgy and evolution, 1994,11:769-777
    [167] Ham RCHJ,Hart HT, Mes THM,et aI.Molecular evolution of noncoding regions of the chloroplast genome in the Crassulaceae and related species.Current Genetics, 1994,25:558-566
    [168] Mes THM, Hart HT.Sedum surculosum and S.jaccardianum share a unique 70 bp deletion in the chloroplast DNA trnL-trnF intergenic spacer.Plant systematics and Evolution, 1994,193:213-221
    [169] Curtis,SE,Clegg MT.Molecul ar evolution of chloroplast DNA sequences .Mol. B ioI.Evol., 1984, 1:291~301
    [170] Germano J,Klein AS.Species-specific muclear and chloroplast single nucleotide polymorphisms to distinguish Picea glauco,Emariana.Theor Appl Genet, 1999,99:37-49
    [171] 卢孟柱,谢红丽等.利用叶绿体DNA变异研究胡杨系统发育.西北植物学报,2000,20(6):1148~1154
    [172] 张文蘅,陈之端,陈虎彪,等.从叶绿体DNA trnL-F序列论双参属的归属问题.植物分类学报,2001,39(4):337~344
    [173] 王玛丽,陈之端,张宪春,等.蹄盖蕨科的系统发育:叶绿体DNA trnL~F区序列证据.植物分类学报.2003,41(5):416~426
    [174] 牟少华.我国部分鸢尾属(Iris)植物系统位置研究[博士学位论文].北京:中国林业科学研究院,2005.79
    [175] Joshi,S. E, V.S.Gupta, R.K.Aggarwal,EK.Ranjekar& D.S.Brar,2000.Genetic diversity and phylogenetic relationship as revealed by inter-simple sequence repeat (ISSR) polymorphism in the genus Oryza.Theor Appl Genet 100:1311~1320
    [176] 钱韦,葛颂,洪德元.采用RAPD和ISSR标记探讨中国疣粒野生稻的遗传多样性.植物学 报,2000,42(7):741~750
    [177] JIANG Jing, YANG Chuan-ping, LIU Gui-Feng ,et al. Genetic relationship of interspeeies for eight birch species. Journal of Forestry Research, 13(4): 281-284 (2002):281~284
    [178] 余爱丽,张木清,陈如凯.ISSR分子标记在甘蔗及其近缘属分类上的应用.福建农林大学学报(自然科学版,2002,31(4):484~489
    [179] 马朝芝,傅廷栋,Stine Tuevesson等.用ISSR标记技术分析中国和瑞典甘蓝型油菜的遗传多样性.中国农业科学,2003,36(11):1403~1408
    [180] 沈颖.石斛类ISSR标记研究与ITS序列分析[硕士学位论文].杭州:浙江大学,2004,28
    [181] 刘小莉,刘飞虎,李宗菊等.10种报春花亲缘关系的ISSR分析.云南大学学报(自然科学版),2004,26(5):454~458
    [182] 邱英雄,胡绍庆,陈跃磊,等.ISSR-PCR技术在桂花品种分类研究中的应用.园艺学报,2004,3(4): 529~532
    [183] LIU Gui-feng, DONG Jing-xiang, JIANG Ying, et al. Analysis of genetic relationship in 12 species of Section Strobus with ISSR markers. Journal of Forestry Research,2005,16(3): 213-215
    [184] 杨淑达,施苏华,龚洵,等.滇牡丹遗传多样性的ISSR分析.生物多样性,2005,13(2):105~111
    [185] 罗瑜萍,龚维,邱英雄,等.羊蹄甲属3种园艺树种分子鉴定及亲缘关系的ISSR分析.园艺学报,2006,33(2):433~436
    [186] 赵杨,陈晓阳,王秀荣,等.9种胡枝子亲缘关系的ISSR分析.吉林林业科技,2006,35(2):1~4
    [187] 沈洁,丁小余,丁鸽,等.铁皮石斛居群差异的研究ISSR指纹标记方法的建立与优化.中国中药杂志,2006,31(4):291~294
    [188] 徐红,李晓波,丁小余,等.中药黄草石斛rDNA ITS序列分析.药学学报,2001,36(10):777~783
    [189] David Tai-wai Lau,Pang-chui Shaw ,Jun wang et al.Authentication of medicinal Dendrobium species by the Internal Transcribed Spacer of ribosomal DNA .Planta Med.,2001,67:456~460
    [190] 丁小余,徐珞珊,徐红,等.曲茎石斛及其近似种鉴别的形态和DNA分子证据.药学学报,2001,36
    [191] 张铭,黄华荣,廖苏梅,等.石斛属RAPD分析及鉴定铁皮石斛的特异性引物设计.中国中药杂志,2001,26(7):442~447
    [192] 丁小余,王峥涛,徐红,等.枫斗类石斛rDNA ITS区的全序列数据库及其序列分析鉴别.药学学报.2002,37(7):567~573
    [193] 罗玉明,丁小余,徐珞珊,等.细叶石斛的位点特异性PCR鉴别.淮阴师范学院学报(自然科学版),2002,1(1):82~85.
    [194] 滕艳芬,吴晓俊,徐红,等.石斜及其常见混淆品的matK基因序列比较.中国药科大学学报,2002,33(4):280~283
    [195] 虞泓,和锐,倪念春,等.石斛属4种植物的AFLP分析.中草药,25(7):808~810
    [196] 王玉东.兰属和石斛属植物的遗传多样性研究[硕士学位论文].杭州:浙江工业大学,2004,34
    [197] 林夏珍,赵建强.中国野生花卉种质资源调查综述.浙江林学院学报,2001,15(4):441~444
    [198] 郑键,张彦广,刘冬云.中国北方野生花卉种质资源研究现状.河北农业大学学报,2002,25(2):22~26
    [199] 卢磊.林木和花卉种质资源科技基础平台建设.[硕士学位论文].北京:北京林业大学,2006,2

© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号

地址:北京市海淀区学院路29号 邮编:100083

电话:办公室:(+86 10)66554848;文献借阅、咨询服务、科技查新:66554700