In vitro Asymbiotic Germination and Propagation of Paphiopedilum concolor (Lindl.) Pfitz.
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摘要: 以授粉后不同发育时期的同色兜兰种子为材料,观察其形态特征和萌发过程,并探讨建立同色兜兰高效快繁体系的最佳条件.结果表明,种子发育中后期即授粉后210~240 d为较适宜的采收期,授粉后210 d的种子萌发率最高(达77.79%);1/4 MS和1/2 MS为同色兜兰适宜的基本培养基,添加100 mL/L椰乳或1 g/L 蛋白胨对种子萌发及原球茎生长和分化有明显的促进作用;添加1 g/L活性炭对原球茎褐化有一定的抑制作用,但添加剂量不宜过大;添加香蕉汁和苹果汁对同色兜兰种子萌发和原球茎生长分化有抑制作用;暗处理对同色兜兰种子萌发无影响;分化后的原球茎在壮苗和生根培养基上培养120 d即可得到4~5片叶、高3~5 cm的同色兜兰健壮试管苗.Abstract: Different embryo-aged Paphiopedilum concolor seeds were collected and their structure, germination process, and effective regeneration conditions were investigated. Results showed that the seeds collected from 210-240 days after pollination (DAP) were optimum for culture in vitro, and the seed germination percentage of 210 DAP was the highest (77.79%). Furthermore, 1/4 MS (Murashige and Skoog culture medium) and 1/2 MS were both suitable for germination, and the addition of 100 mL/L coconut water (CW) and 1 g/L peptone had a positive effect on seed germination and protocorm differentiation. The addition of 1 g/L activated charcoal (AC) inhibited protocorm browning and death, while the addition of 2 g/L AC increased the browning and mortality rates. Banana juice and apple juice had a negative effect on seed germination and protocorm differentiation. After culturing of transferred differentiation protocorm on rooting medium for 120 days, the healthy robust seedlings were 3-5 cm tall and had 5-6 leaves.
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Keywords:
- Paphiopedilum concolor /
- Asymbiotic germination /
- Seed mature /
- Medium
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[1] 陈心启. 中国植物志: 第17卷[M]. 北京: 科学出版社, 1999. Chen XQ. Flora of China: Vol. 17[M]. Beijing: Science Press, 1999.
[2] 罗毅波, 贾建生, 王春玲. 初论中国兜兰属植物的保护策略及其潜在资源优势[J]. 生物多样性, 2003, 11(6): 491-498. Luo YB, Jia JS, Wang CL. Conservation strategy and potential advantages of the Chinese Paphiopedilum[J]. Biodiversity Science, 2003, 11(6): 491-498.
[3] 龙波, 龙春林. 兜兰属植物及其研究现状[J]. 自然杂志, 2006, 28(6): 341-344. Long B, Long CL. Amazing Paphiopedilum and its research status[J]. Chinese Journal of Nature, 2006, 28(6): 341-344.
[4] 刘仲健, 陈心启, 陈利君, 雷嗣鹏. 中国兜兰属植物[M]. 北京: 科学出版社, 2009. Liu ZJ, Chen XQ, Chen LJ, Lei SP. The Genus Paphiopedilum in China[M]. Beijing: Science Press, 2009.
[5] 曾宋君, 陈之林, 吴坤林, 段俊. 国产兜兰属植物的引种和栽培[J]. 中国野生植物资源, 2010, 29(2): 53-58. Zeng SJ, Chen ZL, Wu KL, Duan J. Study on introduction and cultivation of Paphiopedilum distributed in China[J]. Chinese Wild Plant Resources, 2010, 29(2): 53-58.
[6] 曾宋君, 田瑞雪, 陈之林, 吴坤林, 段俊. 兜兰属植物杂交育种研究进展[J]. 热带亚热带植物学报, 2010, 18(2): 459-468. Zeng SJ, Tian RX, Chen ZL, Wu KL, Duan J. Research progress on cross breeding of Paphiopedilum[J]. Journal of Tropical and Subtropical Botany, 2010, 18(2): 459-468.
[7] 李秀玲, 王晓国, 李春牛, 周锦业, 邓杰玲, 曾宋君,卜朝阳, 卢家仕. 基于灰色关联分析方法评价13种野生兜兰的迁地保护适应性[J]. 植物科学学报, 2015, 33(3): 326-335. Li XL, Wang XG, Li CN, Zhou JY, Deng JL, Zeng SJ, Bu ZY, Lu JS. Adaptability evaluation of ex situ conservation of thirteen wild Paphiopedilum species by gray-correlation analysis[J]. Plant Sciense Journal, 2015, 33(3): 326-335.
[8] Arditti J. Micropropagation of Orchids: Vol. 2[M]. New York: Blackwell Publishing, 2008: 49.
[9] Pierik RLM, Sprenkels PA, Van der Harst B, Van der Meys QG. Seed germination and further development of plantlets of Paphiopedilum ciliolare Pfitz. in vitro[J]. Sci Hortic, 1988, 34: 139-153.
[10] Stimart DP, Ascher PD. In vitro germination of Paphiopedilum seed on a completely defined medium[J]. Sci Hortic, 1981, 14: 165-170.
[11] Lee YI, Lee N, Yeung EC, Chung MC. Embryo development of Cypripedium formosanumin relation to seed germination in vitro[J]. Am Soc Hort Sci, 2005, 130(5): 747-753
[12] 张毓, 张启翔, 赵世伟, 凌春英. 濒危植物大花杓兰胚与珠被发育的研究[J]. 园艺学报, 2010, 37(1): 72-76. Zhang Y, Zhang QX, Zhao SW, Ling CY. Embryo and integument development of the endangered species Cypripedium macranthos Sw.[J]. Acta Horticulturae Sinica, 2010, 37(1): 72-76.
[13] Zeng SJ, Zhang Y, da Silva JAT, Wu KL, Zhang JX, Duan J. Seed biology and in vitro seed germination of Cypripedium[J]. Crit Rev Biotechnol, 2014, 34(4): 358-371.
[14] 张娟娟, 严宁, 胡虹. 三种兜兰属植物种子发育过程及其与无菌萌发的关系[J]. 植物分类与资源学报, 2013, 35(1): 33-40. Zhang JJ, Yan N, Hu H. Seed development of three Paphiopedilum species in relation to asymbiotic germination[J]. Plant Diversity and Resources, 2013, 35(1): 33-40.
[15] 丁长春, 虞泓, 刘方媛. 影响杏黄兜兰种子萌发的因素[J]. 云南植物研究, 2004, 26(6): 673-677. Ding CC, Yu H, Liu FY. Factors affecting the germination of Paphiopedilum armeniacum[J]. Acta Botanica Yunna-nica, 2004, 26(6): 673-677.
[16] 陈之林, 叶秀粦, 梁承邺, 段俊. 杏黄兜兰和硬叶兜兰的种子试管培养[J]. 园艺学报, 2004, 31(4): 540-542. Chen ZL, Ye XL, Liang YC, Duan J. Seed germination in virto of Paphiopedilum armeniacum and P. micranthum[J]. Acta Horticulturae Sinica, 2004, 31(4): 540-542.
[17] Lee YI. The asymbiotic seed germination of six Paphiopedilum species in relation to the time of seed collection and seed pretreatment[J]. Acta Hortic, 2007, 75: 381-385.
[18] 周丽, 邓克云, 徐正海. ‘金州报春'兜兰的非共生萌发与快速繁殖[J]. 植物生理学报, 2013, 49(11): 1250-1254. Zhou L, Deng KY, Xu ZH. In vitro asymbiotic germination and propagation of Paphiopedilum primulinum 'Jinzhoubaochun'[J]. Plant Physiology Journal, 2013, 49(11): 1250-1254.
[19] Zeng SJ, Wu KL, da Sliva JAT, Zhang JX, Chen ZL, Xia NH, Duan J. Asymbiotic seed germination, seedling development and reintroduction of Paphiopedilum wardii Sumerh, an endangered terrestrial orchid[J]. Sci Hortic, 2012, 138: 198-209
[20] Lee YI, Yeung EC, Lee N, Chung MC. Embryo development in the lady' 黄玮婷, 曾宋君. 文山兜兰白变种的无菌播种和试管成苗[J]. 植物生理学通讯, 2010, 46(10): 1069-1070. Huang WT, Zeng SJ. Asymbiotic seed germination and in vitro seedling development of Paphiopedilum wenshanense f. album Gruss & Petchl[J]. Plant Physiology Communications, 2010, 46(10): 1069-1070.
[23] 刘其府, 傅燕艳, 曾宋君, 吴坤林, 张建霞, 段俊. 同色兜兰种子非共生萌发试验[J]. 广东农业科学, 2012(12): 47-49. Liu QF, Fu YY, Zeng SJ. Wu KL, Zhang JX, Duan J. Test on asymbiotic seed germination of Paphiopedilum conco-lor (Bateman) Pfitz. 尤佳妍, 张毓, 刘岩, 程瑾, 潘会堂, 张启翔. 小叶兜兰的种子发育和无菌萌发[J]. 植物生理学报, 2014, 50(3): 275-282. You JY, Zhang Y, Liu Y, Cheng J, Pan HT, Zhang QX. Seed development and asymbiotic of Paphiopedilum barbigerum Tang et Wang[J]. Plant Physiology Journal, 2014, 50(3): 275-282.
[25] 王贞, 丛磊, 刘燕. 兜兰属植物研究现状[J]. 林业科学, 2006, 42(7): 114-119. Wang Z, Cong L, Liu Y. A review of Paphiopedilum research[J]. Scientia Silvae Sinicae, 2006, 42(7): 114-119.
[26] 王莲辉, 魏鲁明, 姜运力, 玉屏, 余登利, 潘德权. 白花兜兰的组织培养与快速繁殖[J]. 植物生理学通讯, 2010, 46(10): 1071-1072. Wang LH, Wei LM, Jiang YL, Yu P, Yu DL, Pan DQ. Tissue culture and rapid propagation of Paphiopedilum emersonii Koopowitz et Cribb[J]. Plant Physiology Communications, 2010, 46(10): 1071-1072.
[27] 沈海龙. 植物组织培养[M]. 北京: 中国林业出版社, 2005: 39. Shen HL. Plant Tissue Culture[M]. Beijing: China Forestry Press, 2005: 39.
[28] 曾宋君, 陈之林, 吴坤林, 段俊. 兜兰无菌播种和组织培养研究进展[J]. 园艺学报, 2007, 34(3): 793-796. Zeng SJ, Chen ZL, Wu KL, Duan J. A resume of germination and mericlone of Paphiopedilum[J]. Acta Horticulturae Sinica, 2007, 34(3): 793-796.
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