摘要
本研究为探索珙桐组织培养快速繁殖的有效途径,以珙桐芽为外植体,着重研究各阶段的最佳培养基配方及培养程序,以及培养中试管苗的异常表现及其原因,建立珙桐快繁技术体系。结果表明:
1)利用珙桐芽作为初代培养的外植体材料,最佳的取材时间为10月。不带木质部接种可降低外植体的褐化率。
2)珙桐组织培养条件:启动阶段为昼温26℃,夜温24℃,光照14h/d;继代和生根阶段为昼温24℃,夜温22℃,光照13h/d;光强1500~20001x,pH值为5.8~6.2。
3)珙桐启动培养的最适培养基为1/2MS+BA1.5mg/l+IBA1mg/l,30d后,芽的诱导启动率可达95%以上。
4)愈伤组织诱导丛生芽继代增殖的最适培养基为N_6+BA2.5mg/l+IBA1mg/l+GA_31mg/l,其平均增殖系数可达8.00以上。
5)侧芽继代增殖方式的最适培养基为N_6+BA1mg/l+GA_32mg/l,其平均增殖系数可达3.90。
6)可以试管苗的顶芽和带芽茎段作为外植体,在初代培养基中进行二次启动,启动率高达90%以上,通过增加外植体种类,达到增殖快繁的效果。
7)生根阶段的最适培养基为1/8MS+BA0.1mg/l+NAA1mg/1+活性炭0.5g/1,生根率可达95%以上,生根根数多,生长迅速。
8)炼苗阶段以河沙作为培养基质,并以基本培养基1/10MS为营养液进行叶面施肥,移栽成活率可达62%以上。
The buds of Davidia involucrata were used as explants to establish a rapid technique system of micropropagation, and to study abnormal phenomena of the in vitro shoots and their reasons.
1) Buds were used as explants to establish in vitro system. October was the best period of picking the buds in this test. Removing xylem when inoculating the buds could reduce the browning rate.
2) Temperature for initial culture: Day 26℃, Night 24℃, 14 hours illumination every day; temperature for multiplicative and rooting culture: Day 24℃, Night 22℃, 12 hours illumination every day, and 1500~20001x illumination intensity. The pH of medium was adjusted to 5.8-6.2.
3) The best initiation medium was l/2MS+BA1.5mg/l+IBAlmg/l. After 30 days culture, the inducement rate could be up to 95%.
4) The multiplication of callus induced shoot clumps: N6+BA2.5mg/l+IBAl mg/l+GAslmg/l was the best medium. The multiplication rate of shoot clumps could be up to 8.00.
5) The multiplication of side buds: N6+BAlmg/l+GA32mg/l was the best medium, the multiplication rate of side buds could reach 3.90.
6) The in vitro nodal segments and shoot tips could be used as explants in second initial culture, the inducement rate could reach 90%. A good propagation
result could be obtained by expands the initial explants.
7) The best medium for the rooting of shoots was l/8MS+BA0.1mg/l+NAAl mg/l+AC0.5g/l, in which the root of rooty plantlets grew rapidly and the rooting rate could reach 95%.
8) For nursling of plantlets, sands was used as material and 1/10MS as application of leaves' nutrient solution, the survival rate of plantlets could be up to 62%.
引文
1.傅立国主编.中国植物红皮书.北京:科学出版社.1991:474~475.
2.吴宗福.国家一级保护植物——珙桐.云南林业.1985,4:3~5.
3. Sargent C. S. Plantae Willsonianea. Cambridge: The University Press. 1916,2:255~257.
4.张家勋.中国的鸽子树——珙桐.植物杂志.1998,1:1~7.
5.杨业勤.美丽的中国鸽子树——珙桐.林业科技通讯,1980,9:6~7.
6.张清华,宋朝枢.珙桐天然分布的调查研究.主要珍稀树种繁殖研究.北京:林业出版社.1992,6:45~46.
7.周武忠.花与中国文化.北京:中国农业出版社.1999,184.
8.张家勋,何立明,孔凡伦等.珙桐的形态与生物学特征.北京林业大学学报.1994,16:33~37.
9.贺军辉.蓝果树科3种代表植物的染色体观察.湖南林业科技.1991,2:40~41.
10.孙彬,李柏年.两种珙桐叶片结构的观察.西北植物学报.1993,13:198~202.
11. Hu yushi, Lin Jinxing et al. A comparative study of Davidia involucrate and D. involucrate var. vilimoriniana, International Dendrology Society Year Book(1997).
12.张家勋,李俊清,周宝顺.珙桐的天然分布和人工引种分析.北京林业大学学报.1995,17:25~30.
13.方文培.四川植物志,第一卷.成都:四川出版社.1981.
14.宋朝枢主编.中国珍稀濒危保护植物.北京:中国林业出版社.1989.
15.淘金川等.珙桐的地理分布与引种,浙江林学院学报.1986,1:30~35.
16.王献薄等.珙桐的生物生态学特性和栽培技术.广西植物.1995,15:347~353.
17. Deng Hongbing, Jiang Mingxi et al. Flora and ecological characteristics of rare plant communities on the southern slope of Shennongjia Mountain. Journal of Forestry Research. 2002. 13: 1, 21~24.
18. Yang Y. Q., Xu Y. Y. A preliminary study of the ecological characteristics of dove tree in Guizhou provice Scientia Silvae Sinicae. 1986,22:425~430.
19.彭春良,颜立红.湖南珙桐及其群落的初步研究.湖南林业科技.1989,1:9~13.
20.杨一川,李体俊.四川峨眉山珙桐群落的初步研究,植物生态学与地植物学学报.1989,13:270~276.
21.侯建军.鄂西南长阳县境内珙桐群落的调查分析.湖北民族学院学报:自然科学版.1991,3:94~98.
22.陈波涛,杨成华,洪文武.大方县箐梁子珙桐调查报告.贵州林业科技.1995,23:27~30.
23.焦健.甘肃文县珙桐群落的组成结构数量特征.甘肃农业大学学报.1995,2:189~193.
24.沈泽昊,林洁,陈伟烈等.四川卧龙地区珙桐群落的结构与更新研究.植物生态学报.1999,23:562~567.
25.喻理飞.贵州柏箐喀斯特台原区珙桐群落特征研究.贵州科学.2002,20(3):44~47.
26. Tang, C. Q. Ohsawa, M. Zonal transition of evergreen, deciduous, and coniferous forests along the altitudinal gradient on a humid subtropical mountain, Mt. Emei, Sichuan, China. Plant Ecology. 1997, 133: 1, 63~78..
27. Tang, C. Q. Ohsawa, M. Tertiary relic deciduous forests on a humid subtropical mountain, Mt. Emei, Sichuan, China. Folia Geobotanica & Phytotaxonomica. 2002,37: 1, 93~106.
28.张清华,阎洪.气候变化对我国珍稀濒危树种——珙桐地理分布的影响研究.林业科学.2000,36,(2):47~51.
29.张家勋,李俊清,周宝顺等.珙桐繁殖和栽培技术研究.北京林业大学学报.1995,17:24~28.
30. He, Z. X.; Lao W. X. Evaluation of Chinese tree species in Eastwoodhill Arboretum, Horticulture in New Zealand. 1992,2:7~9.
31. Lamb, J. G. D. Nutty, F. J. The propagation of Davidia. Garden, UK. 1986,111: 4, 189.
32.宋洪初.杭州引种珙桐14年总结.中国植物学会五十周年年会.学术论文摘要汇编.1988,745~746.
33.杨永兰等.珙桐、厚朴、鹅掌楸引种试验研究.重庆林业科技.1993,36:8~10.
34.邓先宝等.珙桐的繁殖措施探讨.重庆林业科技.1993,36:21~25.
35.邹高顺.珍稀树种的种苗繁殖研究.福建林业科技.1994,21:81~88.
36.唐晓军.珙桐的繁育及引种栽培技术.四川林勘设计.2002,1:50~52.
37. Fordham, A. J. Dove tree (Davidia involucrata var. vilmoriniana) and its propagation by seeds. Combined Proceedings - International Plant Propagators' Society. 1988, 37: 343~344.
38.李建强,张敏华,黄宏文等.珙桐的等位酶点变异分析.武汉植物学研究.2000,18:247~249.
39.成俊卿主编.中国热带及亚热带木材.北京:北京科学出版社.1980.
40.黎明,苏金乐,杨芳绒等.珙桐营养器官解剖学研究.河南农业大学学报.1999,33:357~359.
41. Louvier V.; Brouard J. P. et al. n-Alkanes, friedelin and neooleana 3(5) 12 diene in plants. Bulletin de la Societe botanique de France, Letters Botaniques. 1992,139(4~5)307~319.
42. Hang G. Q. Lu F.S. Study on chemical composition of Davidia involucrata Bill. native to China Acta Botanica Sinica. 1989,31:540~543.
43.李轩,陈迎辉,彭春良等.湖南珙桐资源的保护和综合开发模式初探.湖南林业科技.1998,25:45~47.
44. Tsukagoshi, M. Ono, Y. Hashimoto, T. Fossil endocarp of Davidia from the Early Pleistocene sediments of the Tokai Group in Gifu Prefecture, Central Japan. Bulletin of the Osaka Museum of Natural History. 1997,No. 51, 13~23.
45. Manchester, S. R. Leaves and fruits of Davidia (Cornales) from the Paleocene of North America. Systematic Botany. 2002, 27: 2, 368~382.
46.毕世荣,何立明,孔凡伦等.珙桐组织培养.植物生理学通讯.1983,4:43~44.
47.谈锋,刘玉成.缙云山五种珍稀濒危植物的组织培养.重庆林业科技.1993,36:11~15.
48. Sato, T. Microprogation of hankachinoki(Davidia involucrata). Journal of the Japanese Forestry Society. 1996,78:309~311.
49. He, S. A. Jin, W. Jin, J. P. Micropropagation research in the Nanjing Botanic Garden, Mem. Sun Yat-Sen. Botanic Gardens Micropropagation News. 1991, 1: 3, 29~33.
50.周俊辉.植物快速繁殖中存在的问题与对策.仲凯农业技术学院学报.1999,12(4):64~70.
51.李海燕,张丽娟,田和平.细胞和组织培养技术在植物育种中的应用.天津农林科技.1999,10(5):15~16.
52.陈振光主编.园艺植物离体培养学.北京:中国农业出版社.1995,74.
53.于亚军,代汉萍,李宝江.植物激素和生长调节剂在果树组织培养中的应用.北方园艺 2002(6):68~70.
54. Smulder M.J.M,Croes A.F, Wallems G.J, Polar transport of 1-naphthaleneacetic acid determines the distribution of flower buds on explants of tobacco. Plant Physiol. 1988,88: 752~756.
55.谷瑞升,蒋湘宁,郭仲琛等.植物离体培养中器官发生调控机制的研究进展.植物学通报.1999,6(3):238~244.
56.王光萍,黄敏仁.福建山樱花的组织培养及植株再生.南京林业大学学报(自然科学版).2002,26:73~75.
57.卜学贤,陈维纶.活性炭在植物细胞组织培养中的作用.植物学通报.1988,5(1):1~5.
58.卜学贤,陈维纶.活性炭对培养基中植物生长调节物质的吸附作用.植物学通报.1988,14(4):401~405.
59. Steinitz B., Yahel H. In vitro Proapagation of Narcissus tazetta. Hortscience. 1982,17 (3): 333~334.
60.曹孜义,刘国民.实用植物组织培养技术教程.甘肃,甘肃科学技术林业出版社.2001,58.
61.肖玉兰,张立力,张光怡等.非洲菊无糖组织培养的应用研究.园艺学报.1988,25(4):408~410.
62.李孟超.火鹤组培苗无糖生根培养试验.北京农业科学.2000,18(5):30~32.
63.周岩等.植物组织培养中的体细胞无性系变异.河南职业技术师范学院学报.2000,12(4):25~27.
64.梁一池,杨华.植物组织培养技术的研究进展.福建林学院学报.2002,22(1):1~3.
65.谭文澄,戴策刚.观赏植物组织培养技术.北京:中国林业出版社.1991,106~107.
66.李浚明.植物组织培养教程.北京:中国农业大学出版社.1992,360.
67.陈正华.木本植物组织培养及其应用.北京:高等教育出版社.1986,28~29.
68.毕世荣等.桫椤组织培养的研究.植物生理学通讯.1985,(6):38.
69.曹效东,曹孜义.植物试管繁殖的成本和效益浅析.植物生理学通讯.1996,32(4):284~291.
70.蔡新声.台湾植物组织培养研究现状.植物生理学通讯.1994,30(6):473~476.