哥伦比亚花烛的组织培养研究
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摘要
以哥伦比亚花烛四个品种的叶片、叶柄为外植体，研究了各种不同因素对无菌离体培养每一阶段的影响，内容包括：光照、大量元素不同配比、外植体切割方式等因素对愈伤组织诱导、增殖及不定芽分化的影响。
结果表明：愈伤组织诱导应在暗培养下进行，改良后的1／2MS C具有最好的诱导效果，1／2MS C+H_2对四个品种均具有较好的诱导效果。1／2MSC+H_3(B1)对四个品种愈伤组织的不定芽分化均具有较好效果。以与B1成分相同但不添加琼脂的液体培养基对愈伤组织进行再继代，愈伤组织分化出大量不定芽并很快长成叶片。生根培养以1／2MS+NAA0.1mg/L(R2)更为适合。在以椰壳粉：细沙：珍珠岩=1：1：1的混合基质上，在注意保湿保温的条件下，移栽苗全部成活。
同时，以‘Arena’为材料，探索了无菌苗经扩繁再发育成苗并进行移栽的全过程。1／2MS C+BA1.0mg／L(AN3)具有较好的愈伤组织诱导和增殖效果。液体浅层静置培养同样诱导出大量不定芽。
此外，对外植体褐变的遏制进行了一定的研究。认为：外植体消毒方式、外植体切割方式、PVP均对此有影响。其机理和遏制褐变的最佳途径有待进一步研究。
Leaves and stalks are used as explants in the studies on the effect of different factors on nurture in vitro of various cultured varieties of Anthurium andraeanum Lindl.
The content of result includes light , different macro-element portion , cutting way of expiants ; these factors affect inductive rate of callus.
The results are as follows :
Callus should be inducted under darlmess. The best inductive effect belongs to 1/2MS C ( which is the basal media ameliorated from 1/2MS). 1/2MS C+H2 has better effect on four cultured varieties.
1/2MS C+H2 ( Bl ) has better effect on adventitious bud's proliferation OP four cultured varieties. Liquid media same as Bl but Agar powder not added , which used to subculture callus , made adventitious buds proliferated on the callus numerously.
Rooting culture is better on the media l/2MS+NAA0.1mg/L (R1 ).
Using mixture ( cocos nut powder : sand : pearl rock = 1 : 1 : 1 ) as basal matter , paying attention to maintain temperature and humidity , transplanted plantlets are all lively.
The research that the whole course about proliferation of 'Arena' 's plantlet , which is cultivated by method of tissue culture without bacterium , was undertaking at the same time.
1/2MS C+BA1.0mg/L ( AN3 ) has the better effect on callus induction as well as proliferation of adventitious buds. Liquid shallow layer static nurture can also proliferate numerous adventitious buds.
Otherwise , the research of restraining explants' browning has been done in limited level. Disinfecting way of explants , cutting way of explants , PVP all have effect on explants' browning. The research should be done deeply.
结果表明：愈伤组织诱导应在暗培养下进行，改良后的1／2MS C具有最好的诱导效果，1／2MS C+H_2对四个品种均具有较好的诱导效果。1／2MSC+H_3(B1)对四个品种愈伤组织的不定芽分化均具有较好效果。以与B1成分相同但不添加琼脂的液体培养基对愈伤组织进行再继代，愈伤组织分化出大量不定芽并很快长成叶片。生根培养以1／2MS+NAA0.1mg/L(R2)更为适合。在以椰壳粉：细沙：珍珠岩=1：1：1的混合基质上，在注意保湿保温的条件下，移栽苗全部成活。
同时，以‘Arena’为材料，探索了无菌苗经扩繁再发育成苗并进行移栽的全过程。1／2MS C+BA1.0mg／L(AN3)具有较好的愈伤组织诱导和增殖效果。液体浅层静置培养同样诱导出大量不定芽。
此外，对外植体褐变的遏制进行了一定的研究。认为：外植体消毒方式、外植体切割方式、PVP均对此有影响。其机理和遏制褐变的最佳途径有待进一步研究。
Leaves and stalks are used as explants in the studies on the effect of different factors on nurture in vitro of various cultured varieties of Anthurium andraeanum Lindl.
The content of result includes light , different macro-element portion , cutting way of expiants ; these factors affect inductive rate of callus.
The results are as follows :
Callus should be inducted under darlmess. The best inductive effect belongs to 1/2MS C ( which is the basal media ameliorated from 1/2MS). 1/2MS C+H2 has better effect on four cultured varieties.
1/2MS C+H2 ( Bl ) has better effect on adventitious bud's proliferation OP four cultured varieties. Liquid media same as Bl but Agar powder not added , which used to subculture callus , made adventitious buds proliferated on the callus numerously.
Rooting culture is better on the media l/2MS+NAA0.1mg/L (R1 ).
Using mixture ( cocos nut powder : sand : pearl rock = 1 : 1 : 1 ) as basal matter , paying attention to maintain temperature and humidity , transplanted plantlets are all lively.
The research that the whole course about proliferation of 'Arena' 's plantlet , which is cultivated by method of tissue culture without bacterium , was undertaking at the same time.
1/2MS C+BA1.0mg/L ( AN3 ) has the better effect on callus induction as well as proliferation of adventitious buds. Liquid shallow layer static nurture can also proliferate numerous adventitious buds.
Otherwise , the research of restraining explants' browning has been done in limited level. Disinfecting way of explants , cutting way of explants , PVP all have effect on explants' browning. The research should be done deeply.
引文
1 北京林业大学园林系花卉教研组．花卉学(第一版)．北京：中国林业出版社，1994．487～488
2 北京市花卉研究所．室内花卉—新引进的国外观叶植物(第一版)．北京：中国经济出版社，1989．128～130
3 穆鼎．鲜切花周年生产(第一版)．北京：中国农业科技出版社，1997．183～192
4 王宏志．中国南方花卉(第一版)．北京：金盾出版社，1998．42～43
5 何清正．花卉生产新技术(第一版)．广州：广东科技出版社，1991．376～377
6 Adelneid R. Kuehnle, N. Sugii .Induction of Tumors in Anthurium andraeanum by Agrobacterium tumefaciens. HortScience, 1991, 26(10):1325～1328
7 Fure-Chyi chen, Adelneid R. Kuehnle .Obtaining Transgenic Anthurium through Agrobacterium-mediated Transformation of Etiolated Internodes. Journal of A merican Society Horticultural Science, 1996,121(1):47～51
8 Kuehnle A. R., Fure-Chyi chen. Agrobacterium-mediated transformation of Anthurium. In: Bajaj Y. P. S. (ed). Biotechnology in Agriculture & Forestry-Plant Protoplast and Genetic Engineering V. Berlin: Springer Verlag, 1994. Vol 29: 215～225
9 Geier T.. Anthurium .In: P. V. Ammirato, D. A. Evans, W. R. Sharp (eds). Handbook of plant cell culture. New York: McGraw-Hill ,1990.228～252
10 Pierik R. L. M., Steegmans H. H. M., Van der Meys. J. A. J..Plantlet formation in callus tieeues of Anthurium andraeanum Lind..Scientia Horticulturae 1974, 2:193～198
11 Pierik R. L. M..Callus multiplication of Anthurium andraeanum Lind. In
liquid media .Netherlands Journal of Agricultural Science, 1975, 23:299～302
12 Pierik R. L. M.. Anthurim andraeanum Lind. Plantlets Produced from in virto cultivated callus tissues. Physiol. Plant., 1976, 37(1):80～82
13 Pierik R. L. M., Steegmans H. H. M.. Vegetative propagation of Anthurium scherzerianum Schott through callus cultures. Scientia Horticulturae, 1976, 4:291～292
14 Pierik R. L. M., Van leenuwen P., Righter G. C. C. M..Regeneration of leaf explants of Anthurium andraeanum Lind. in virto .Netherlands Journal of Agricultural Science, 1979, 27:221～226
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16 Rosario T. L., Lapitan L. A.. Callus and plantlet formation in Anthurium anraeanum Lind.. Philipp. Agri., 1981, 64:197～202
17 Larkin P. J., Scowcroft W. R.. Somaclonal variation: A novel source of variability from cell cultures for plant improvement. Theor. Appl. Genet., 1981, 60:197～214
18 Finnie J. F., Van Staden J..In virto culture of Anthurium andraeanum Lind.. S. Afr. J. Bot., 1986, 52:343～346
19 Kunisaki J. T.. Tissue culture of tropical ornamental plants. HortScience, 1977, 12:141～142
20 Kunisaki J. T. In virto propagation of Anthurium andaeanum Lind.. HortScience, 1980, 15:508～509
21 Keller E. R. J., Brehmer M., H(?)fer E..Micropropagation of Anthurium andraeanum Lind. and the use of a novel stabilizing substrate. Arch. Gartenbau, 1986, 34:149～156
22 Geier T.. Micropropagation of Anthurium scherzerianum: Propagation
schemes and plant conformity. Acta Horticulturae, 1987, 212:293～298
23 Leffring L., Soede A. C.. Nieuwe vermeerderingsmethode bij weefselkweek Anthurium andraeanum. Vakbald Bloemisterij, 1978, 33(34): 19～20
24 Leffring L., Soede A. C.. Weefselkweek Anthurium andraeanum onderzoek te boven (1). Vakbald Bloemisterij, 1979, 34(13):43
25 Leffring L., Soede A. C..Weefselkweek Anthurium andraeanum onderzoek te boven (2). Vakbald Bloemisterij, 1979, 34(15):40～41
26 Adelheid R. Kuehnle, Fure-Chyi Chen, Nellie Sugii .Somatic embryogenesis and plant regeneration in Anhturium andraeanum hybrids. Plant Cell Reports ,1992,438～442
27 Tracie K. Matsumoto, David T. Webb, Adelheid R. Kuehnle .Histology and Origin of Somatic Embryos Derived from Anthurium andraeanum Linden ex Andr(?) Lamina. J. Amer. Soc. Hort. Sci., 1996, 121(1):404～407
28 Kamemoto H., A. Kuehnle, J. Kunisaki .Breeding for new and bacterial blight resistant cultivars. In: K. M. Delate and C. H. M. Tome (eds). Proc. 5th Hawaii Anthurium Ind. Conf..Honolulu: Univ. Hawaii, 1994. 39～44
29 Fure-Chyi Chen, Adelheid R. Kuehnle, Nellie Sugii .Anthurium roots for micropropagation and Agrobacterium tumefaciens-mediated gene transfer. Plant Cell, Tissue and Organ Culture, 1997, 49:71～74
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2 北京市花卉研究所．室内花卉—新引进的国外观叶植物(第一版)．北京：中国经济出版社，1989．128～130
3 穆鼎．鲜切花周年生产(第一版)．北京：中国农业科技出版社，1997．183～192
4 王宏志．中国南方花卉(第一版)．北京：金盾出版社，1998．42～43
5 何清正．花卉生产新技术(第一版)．广州：广东科技出版社，1991．376～377
6 Adelneid R. Kuehnle, N. Sugii .Induction of Tumors in Anthurium andraeanum by Agrobacterium tumefaciens. HortScience, 1991, 26(10):1325～1328
7 Fure-Chyi chen, Adelneid R. Kuehnle .Obtaining Transgenic Anthurium through Agrobacterium-mediated Transformation of Etiolated Internodes. Journal of A merican Society Horticultural Science, 1996,121(1):47～51
8 Kuehnle A. R., Fure-Chyi chen. Agrobacterium-mediated transformation of Anthurium. In: Bajaj Y. P. S. (ed). Biotechnology in Agriculture & Forestry-Plant Protoplast and Genetic Engineering V. Berlin: Springer Verlag, 1994. Vol 29: 215～225
9 Geier T.. Anthurium .In: P. V. Ammirato, D. A. Evans, W. R. Sharp (eds). Handbook of plant cell culture. New York: McGraw-Hill ,1990.228～252
10 Pierik R. L. M., Steegmans H. H. M., Van der Meys. J. A. J..Plantlet formation in callus tieeues of Anthurium andraeanum Lind..Scientia Horticulturae 1974, 2:193～198
11 Pierik R. L. M..Callus multiplication of Anthurium andraeanum Lind. In
liquid media .Netherlands Journal of Agricultural Science, 1975, 23:299～302
12 Pierik R. L. M.. Anthurim andraeanum Lind. Plantlets Produced from in virto cultivated callus tissues. Physiol. Plant., 1976, 37(1):80～82
13 Pierik R. L. M., Steegmans H. H. M.. Vegetative propagation of Anthurium scherzerianum Schott through callus cultures. Scientia Horticulturae, 1976, 4:291～292
14 Pierik R. L. M., Van leenuwen P., Righter G. C. C. M..Regeneration of leaf explants of Anthurium andraeanum Lind. in virto .Netherlands Journal of Agricultural Science, 1979, 27:221～226
15 Pierik R. L. M.. Vermeerdering in kweekbuizen stijgt tot 30 miljeon. Vakblad voor de Bloemisterij, 1985,40 (26):44～45
16 Rosario T. L., Lapitan L. A.. Callus and plantlet formation in Anthurium anraeanum Lind.. Philipp. Agri., 1981, 64:197～202
17 Larkin P. J., Scowcroft W. R.. Somaclonal variation: A novel source of variability from cell cultures for plant improvement. Theor. Appl. Genet., 1981, 60:197～214
18 Finnie J. F., Van Staden J..In virto culture of Anthurium andraeanum Lind.. S. Afr. J. Bot., 1986, 52:343～346
19 Kunisaki J. T.. Tissue culture of tropical ornamental plants. HortScience, 1977, 12:141～142
20 Kunisaki J. T. In virto propagation of Anthurium andaeanum Lind.. HortScience, 1980, 15:508～509
21 Keller E. R. J., Brehmer M., H(?)fer E..Micropropagation of Anthurium andraeanum Lind. and the use of a novel stabilizing substrate. Arch. Gartenbau, 1986, 34:149～156
22 Geier T.. Micropropagation of Anthurium scherzerianum: Propagation
schemes and plant conformity. Acta Horticulturae, 1987, 212:293～298
23 Leffring L., Soede A. C.. Nieuwe vermeerderingsmethode bij weefselkweek Anthurium andraeanum. Vakbald Bloemisterij, 1978, 33(34): 19～20
24 Leffring L., Soede A. C.. Weefselkweek Anthurium andraeanum onderzoek te boven (1). Vakbald Bloemisterij, 1979, 34(13):43
25 Leffring L., Soede A. C..Weefselkweek Anthurium andraeanum onderzoek te boven (2). Vakbald Bloemisterij, 1979, 34(15):40～41
26 Adelheid R. Kuehnle, Fure-Chyi Chen, Nellie Sugii .Somatic embryogenesis and plant regeneration in Anhturium andraeanum hybrids. Plant Cell Reports ,1992,438～442
27 Tracie K. Matsumoto, David T. Webb, Adelheid R. Kuehnle .Histology and Origin of Somatic Embryos Derived from Anthurium andraeanum Linden ex Andr(?) Lamina. J. Amer. Soc. Hort. Sci., 1996, 121(1):404～407
28 Kamemoto H., A. Kuehnle, J. Kunisaki .Breeding for new and bacterial blight resistant cultivars. In: K. M. Delate and C. H. M. Tome (eds). Proc. 5th Hawaii Anthurium Ind. Conf..Honolulu: Univ. Hawaii, 1994. 39～44
29 Fure-Chyi Chen, Adelheid R. Kuehnle, Nellie Sugii .Anthurium roots for micropropagation and Agrobacterium tumefaciens-mediated gene transfer. Plant Cell, Tissue and Organ Culture, 1997, 49:71～74
30 Chen F. C..Genetic engineering of anthurium for bacterial disease resistance.[PhD. diss.]. Honolulu: Univ. Hawaii. 1993
31 D. J. Norman, A. M. Alvarez. Latent infections of in vitro anthurium caused by Xanthomonas campestris pv. dieffenbachia. Plant Cell, Tissue and Organ Culture, 1994, 39:55～61
32 Arnold H. Hara , Wayne T. Nishijima , James D. Hansen . Reduced Pesticide Use in an IPM Program for Anthuriums . Journal of Economic
Entomology, 1990, Vol.83(4): 1531～1534
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