大花蕙兰组织培养技术体系研究
详细信息    本馆镜像全文|  推荐本文 |  |   获取CNKI官网全文
摘要
本研究为探索大花蕙兰(Cymbidium grandifiorium)组织培养快速繁殖的有效途径,以大花蕙兰的茎尖为外植体,着重研究其培养的最佳培养基配方及培养程序,以及培养中防止氧化褐变的措施,建立大花蕙兰快繁技术体系。结果表明:
     1)大花蕙兰组织培养条件为:昼温25±2℃,夜温23±2℃,光照14H/D,光强20001x左右,pH值为5.3~5.6。
     2)启动培养:启动培养基为MS+BA1.0mg/L+NAA0.01mg/L+AC0.5g/L效果最好,25天后启动率可达85.7%。
     3)丛生芽增殖:最适培养基为MS+BA2.0mg/L+NAA0.1mg/L+AC0.5g/L,丛生芽增殖最快,80天增殖系数可达6.23。
     4)PLB诱导:培养基为1/2MS+BA1.0mg/L+NAA0.1mg/L+AC0.1g/L诱导效果最好,诱导率可达71.4%。
     5)PLB增殖:培养基为1/2MS+BA1.0mg/L+NAA0.05mg/L+AC0.1g/L增殖倍数最高,90天时可达50。
     6)成苗阶段:促进苗生长最适培养基为1/2MS+BA2.0mg/L+NAA0.1mg/L+AC0.58/L。促进根生长最适培养基为1/2MS+BA1.0mg/L+NAA0.1mg/L+AC0.5g/L。
     7)AC对外植体褐变的防止有一定作用,浓度为0.58/L启动培养效果最优。在PLB诱导和增殖阶段则以0.1g/L浓度效果较优。
     8)炼苗以竹林下的腐叶土为基质效果最好,小苗成活率可达100%。
The shoots of Cymbidium grandiflorium were used as explants to establish a rapid technique system of micropropagation, and to find out effective measures to prevent the browning of shoot explants. The results were as follows:
    1) Temperature for culture: Day 25℃, Night 23℃. 14 hours illumination every day, and about 2000 Lux illumination intensity. The pH of medium was adjusted to 5.3-5.6.
    2)The best initiation medium was MS+BA1.0mg/L+NAA0.01mg/L+AC0.5g/L. After 25 days culture, the rate of initiation could reach 85.7%.
    3)The multiplication of shoot clumps:MS+BA2.0mg/L+NAA0.1mg/L+AC0.5g/L was the best medium. After 80 days culture, the multiplication rate of shoot clumps could reach 6.23.
    4) The inducement medium of the PLB: l/2MS+BA1.0mg/L+NAA0.1mg/L+ ACO.lg/L. After 80 days culture, the inducing rate of the PLB could reach 71.4%.
    5) The multiplication of the PLB:l/2MS+BA1.0mg/L+NAA0.05mg/L+AC0.1g/L was the best medium. After 90 days culture, the multiplication rate of the PLB could reach 50.
    6) Seedling stage: the medium for the elongation of shoots and leaves was l/2MS+BA2.0mg/L+NAA0.1mg/L+AC0.5g/L. the medium for the rooting of shoots was 1/2MS+BA1.0mg/L+NAA0.1mg/L+AC0.5g/L.
    
    
    7) Active carbon was effective for preventing the explants from browning to a certain extent, 0.5g/L was the best initiation concentration, 0.1 g/L was the best inducement and multiplication the PLB concentration.
    8) For nursling of plantlets, the soil of the rotten leaves in the bamboo groves was the best material, the survial rate of plantlets could be up to 100%, after transplanting soil.
引文
1.卢思聪.中国兰与洋兰.北京:金盾出版社,1994
    2.谢为龙,谭群英,王爱平.影响大花蕙兰试管苗培育的因素研究.四川农业大学学报,1994,12(2):231~234
    3.王熊.兰花的组织培养.见:罗士韦,许智宏主编 经济植物的组织培养.科学出版社,1989,170~175
    4.卢思聪.兰花栽培入门,北京:科学出版社,1988
    5.李哞.兰之胚培养,中国园艺(台),1990,36(4):223~244
    6.陈刚,张远能.中国兰花茎尖组织培养研究,花卉,1998,71(1):13
    7.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):111~116
    8.松华,大花蕙兰史略,花卉,1995,53(1):14
    9.卢思聪,潜力.华贵夺目一派雍容的大花蕙兰,中国花卉报
    10.谷祝平.洋兰—艳丽神奇的世界.成都:四川科学出版社,1991
    11.邬秉左.大花蕙兰及其栽培管理,花卉,1997,67(3):12~13
    12.陈心启.吉占和编著.大花蕙兰——杂交品种系列,中国兰花全书,100~102,中国林业出版社,2000
    13.陈翼华,张志刚,高青等.大花蕙兰的离体繁殖.www.google.com
    14.吴连星,大花蕙兰的栽培管理技术.亚热带植物科学,2001,30(1):54~58
    15.《绿生活》杂志编辑部编辑.东亚兰.最新兰花栽培指南,中国农业出版社,2001,108~117
    16.Morel G. producing vires-free Cymbidiums[J]. Am Orchid Soc Bull, 1960, 29:495~497
    17.Winder D D. Clonal multiplication of Cymbidium through tissue culture of the shoot meristem[J]. Am Orchid Soc Bull, 1963, 32:105~107
    18.Murashige, T. and F Skoog. A revised medium for rapid growth and bioassays with tobacco tissue cultures[J]. Physiol.Plant, 1962, 15:473~479
    19.Rao, A. N. 1977, Tissue culture in the orchid industry.. Edited by J.Reinert and Y..S.P. Bajaj.(eds). Applied and Fundamental Aspects of Plant Cell Tissue and Organ Culture. Berlin:. Springer Verlag, 1977.44
    20.Arditti, J.Clonal propagation of orchids by means of tissue culture. In Arditti J(ed). Orthid Biology. Reviews and Perspective, I. Ithaca, NewYork: Comell University Press Ltd., 1977.203
    
    
    21.陈维伦.我国植物快速繁殖和无毒种苗生产的现状和问题见:植物生物技术改良.孙敬山,陈维伦主编.北京:中国科技出版社,1991.213
    22.罗士韦.植物组织与细胞培养研究工作的进展.全国细胞培养与体细胞杂交线粒体呼吸代谢与杂种优势会议文集.1978.1
    23.Kee-Youep Pack, Lu-Kwang, Bong-hee Han. Perspective and handicaps for commericial application of micropropagtion in Korea. Advances in Development Biology and Biotechology of higher Plants. Plant Tissue Culture. Published Korea Soc, 1993. 38
    24.曹孜义主编.实用植物组织培养技术教程.兰州:甘肃科学出版社,2001
    25.孙安慈,建兰根状茎增殖条件的研究.植物学通报,1989,6(3):147~150
    26.王熊,陈季楚,刘桂云等.建兰和秋兰原球茎的发生及无性系的建立.植物学报,1981,(7):203~207
    27.毛碧增,林蔚红,钱秀红等.影响建兰原球茎增殖的若干因素.浙江农业大学学报,1998,24(1):66~68
    28.张菊野,俞玲凤,连宏坤.几种影响春兰原球茎生长与分化的因素.植物生理学通讯,1993,29(3):175~178
    29.曾宋君,程式君,张京丽等.墨兰及其杂种的组织培养与快速繁殖.广西植物,1998,18(2):153~156
    30.张志胜,欧秀娟.墨兰的组织培养.园艺学报,1995,22(3):303~304
    31.王衍安,徐瑛,王志武.培养条件对墨兰组培芽增殖和生长的影响.山东林业科技,1999,121(2):15~17
    32.吴汉珠,王续衍,林泰碧.‘中国兰’茎顶组织培养研究.园艺学报,1987,14(3):203~207
    33.郭达初,刘克斌,柴明良等.蝴蝶石斛兰、卡特兰、蝴蝶兰离体快速繁殖的研究.浙江农业学报,1991,3(1):34~38
    34.王怀宇.蝴蝶兰的快速无性繁殖.园艺学报,1989,16(1):73~77
    35.杨美纯,周歧伟,许鸿源等.外部因子对蝴蝶兰原球茎状体发生的影响.广西植物,2000,20(1):42~46
    36.李进进,廖俊杰,柯丽婉等.蝴蝶兰根段的组织培养.植物生理学通讯,2000,36(1):37
    
    
    37.张秀清,王志武,王春英等.蝴蝶兰实生苗原球茎诱导研究.莱阳农学院学报,1995,12(1):44~46
    38.傅连芳等.蝴蝶兰快速繁殖.热带作物研究,1990,(2):24~27
    39.颜东敏.蝴蝶兰叶片愈伤组织繁殖的探讨,兰花世界,1989,75~79
    40.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
    41.杨其光,杜国华.霍山石斛嫩枝离体培养成苗.植物生理学通讯,1986,12
    42.唐雪梅,陈红.激素对曲茎石斛试管苗生长的影响.中药材,1993,16(8):12~13
    43.曾宋君,程式君.石斛的试管苗快速繁殖.中药材,1996,19(10):490~491
    44.杨联河,王倩嵘,石拓等.曲茎石斛组织培养研究.中国中药杂志,1998,23(11):658~659
    45.王光远,许智宏,蔡德发等.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
    46.叶秀粪.石斛兰组织培养和细胞学观察.园艺学报,1995,22(1):83~87
    47.张菊野等.卡托利亚兰的组织培养.植物生理学通讯,1988,(5):51
    48.彭晓明,曾宋君,张京丽等.文心兰的茎尖及花梗组织培养和快速繁殖.园艺学报,2000,27(2):127~129
    49.陈兴贻.文心兰的组织培养.植物生理学通讯,1989,(6):49
    50.何松林,十三鸟和子,孔德政等.基本培养基及凝固剂对文心兰试管苗生长发育的影响.北京林业大学学报,2001,23(1):29~31
    51.朱根发,王怀宇,陈明莉.文心兰脱毒快速繁殖技术研究.广东农业科学,1997,(4):27~28
    52.谭文澄.观赏植物组织培养技术.北京:中国林业出版社,1991,237~247
    53.Sagawa Y, Sengal O P. Aseptic stem propagation of Vanda miss Juaqum[J]. Lindleyana. 1988, (3): 27~29
    54.Sigh F, Prakash D. Proceeding of a National Symposium on Plant Propagtion[M]. Indian: Calcatta West Bengal, 1982. 103~104
    55.Bhojwani S S, Razdan M K. Plant Tissue Culture: Theory and Practice[M]. Amsterdam_Oxford_New York Tokyo, 1977
    56.Vij S P, Anil Sood, Plaha K K. Propgation of Rhynchostylis Retusa BL. (Orchidaceae) by direct organogensis from leaf segment culture[J].Bot Goz, 1984, 145(2): 210~214
    
    
    57.孙安慈.兰花组培工作进展简介。植物杂志,1988,(6):24~25
    58.Mroginski L A, Kartha K K, Shyluk J P. Regeneration of Peanut(Arachis hypogea) plantlets by in vitro culture of immature leaves[J].Can J Bot, 1981, 59(5): 826~830
    59.Seeni S, Latha P G. Foliar regeneration of the endangered Red vanda, Renanthera imschootiana Rolfe[J]. Am Orchid Soc Bull, 1976, 45:1022~1024
    60.谷祝平,颜廷进,大花蕙兰茎尖组织培养及其形态建成研究,实验生物学报.1989,22(2):149~151
    61.郑迎冬,杨承勇,蒋林.大花蕙兰的茎段培养.仲恺农业技术学院学报,2000,13(1):19~22
    62.刘明志,朱京育.培养基、BA 和复合添加物对大花蕙兰增殖和分化的影响.暨南大学学报(自然科学版),2000,21(3):100~105
    63.张树珍,曾宪松,张银东等.大花蕙兰的组织培养.热带作物研究,1995,(4):26~28
    64.肖三元.金线兰组织培养及快速繁殖研究.广西热作科技,1998,66(1):12~14
    65.王亦菲,陆瑞菊,周润梅等.丛生芽——文心兰快速繁殖的新途径.66~67 朱德蔚主编.植物组织培养与脱毒快繁技术,中国科学出版社,2001,10
    66.刘荣维,梅庆超,崔元方等.丛生芽——蝴蝶兰无性快速繁殖的新途径.热带作物学报,1993,14(2):105~107
    67.周俊彦,郭扶兴.植物快速营养繁殖速度及产率的计算.植物学通报,1991,8(2):60~63
    68.李云主编.林果花菜组织培养快速育苗技术.中国林业出版社,2001,123
    69.赵杨景,郭顺星,高薇薇等.3 种内生真菌与大花蕙兰共生对矿质营养吸收的影响.园艺学报,1999,26(2):110~115
    70.Dan-hua Yu,Carole P Mereclith. The influence of explant origin on tissue browning and shoot production in shoottip cultures of grapevine. J. Amer. Soc. Hort Sci .1986(16): 972~975
    71.王熊.兰花快速无性繁殖的研究及花芽分化的探讨.植物生理学报,1984,10:391~396
    72.王熊.兰花生长发育规律的探讨.山东农业大学学报,1988,19(1):19~24
    73.王光远,许智宏,蔡德发等.铁皮石斛的离体开花.中国科学,1996,37(5):229~234

© 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