早开堇菜组织培养及植株再生体系的建立

doi:10.11686/cyxb2015160

摘要/Abstract

摘要： 以野生早开堇菜为材料,研究了不同外植体类型(子叶节、叶片、叶柄)和不同激素配比对其不定芽诱导、愈伤组织诱导和分化的影响,成功建立了早开堇菜组织培养植株再生体系。结果表明,1) 诱导子叶节和叶柄不定芽诱导的最适培养基为 MS+2.0 mg/L 6-BA+0.1 mg/L NAA,叶片不适合作为直接诱导不定芽的外植体。2) 3种外植体均能诱导愈伤组织,子叶节愈伤组织诱导的时间最短,其次为叶柄,叶片最晚。子叶节和叶柄愈伤组织诱导的最适培养基为MS+1.0 mg/L 6-BA+0.5 mg/L 2,4-D,诱导率分别为98.3%和96.7%;叶片愈伤组织诱导的最适培养基为MS+1.0 mg/L 6-BA+0.5 mg/L 2,4-D+0.05 mg/L NAA,诱导率可达88.3%。3) 最适愈伤组织分化培养基为:MS+1.5 mg/L 6-BA+0.1 mg/L NAA,分化率为100%。4)不定芽增殖的最佳培养基为:MS+1.5 mg/L 6-BA+0.075 mg/L NAA,增殖率为4.68。 5)再生苗移植到添加1.0 mg/L 6-BA的1/2 MS培养基上,生根率92.3%。6)组培苗移栽到珍珠岩∶河沙∶腐殖质(1∶2∶2)的混合基质时,100%成活,且植株长势好。

Abstract: The effects of various hormone ratios on adventitious buds, callus inductions and differentiation were investigated using the cotyledon node, leaf blade and petiole of Viola prionantha as explants. Finally, a plant regeneration system via tissue culture was established. The results showed that the optimal medium for adventitious bud induction from the cotyledon node and petiole was MS+2.0 mg/L 6-BA+0.1 mg/L NAA,but the leaf blade was not suitable as an explant. Three kinds of explants could induce callus formation. The optimal medium for callus induction from the cotyledon node and petiole was MS+1.0 mg/L 6-BA+0.5 mg/L 2,4-D with the highest callus induction rate being 98.3% and 96.7%, respectively. The optimal medium for callus induction from the leaf blade was MS+1.0 mg/L 6-BA+0.5 mg/L 2,4-D+0.05 mg/L NAA with the highest callus induction rate being 88.3%. The best medium for callus differentiation was MS+1.5 mg/L 6-BA+0.1 mg/L NAA with the highest callus differentiation rate being 100%. The optimal medium for the proliferation of adventitious buds was MS+1.5 mg/L 6-BA+0.075 mg/L NAA,the highest proliferation times being 4.68. The rooting rate of regenerated plants was up to 92.3% on half strength MS medium supplemented with 1.0 mg/L 6-BA. Rooted plantlets were grown on mixture with perlite: sand: humus (1:2:2); survival rate was up to 100% with good growth.

李俊强, 林利华, 张帆, 万雪琴, 刘敏, 赵景龙. 早开堇菜组织培养及植株再生体系的建立[J]. 草业学报, 2015, 24(11): 163-173.

LI Jun-Qiang, LIN Li-Hua, ZHANG Fan, WAN Xue-Qin, LIU Min, ZHAO Jing-Long. Tissue culture in vitro and plant regeneration of Viola prionantha[J]. Acta Prataculturae Sinica, 2015, 24(11): 163-173.

参考文献

[1] Wang L N. The Life Cycle and Ecological Adaptation of Viola Prionantha Bunge[D]. Hohhot: Inner Mongolia University, 2009.
[2] Liu H C, Sun Z Y. Study on the introduction and application of three kinds of wild genus Viola . Chinese Landscape Architecture, 2006, 1(13): 1-4.
[3] Zhang J Z, Sun G F, Liu X C. Available herbaceous flower resources in greening of environmental for the city of Beijing. Proceedings of Beijing Olympics and the City Landscape Construction[C]. Beijing: Beijing Institute of Landscape Architecture, 2002.
[4] Zhou C. The Textual Research and the Studies on the Chemical Constituents of Viola yedoensis [D]. Beijing: Beijing University of Chinese Medicine, 2008: 13-15.
[5] Liu X X, Liu J H, Liu Z K, et al . Studies on effective composition analysis of Viola manshuricaw and its antibacterial effect. Journal of Traditional Chinese Veterinary Medicine, 2004, 23(3): 16-18.
[6] Chen H Y. Determination of volatile components in Viola prionantha Bunge. Journal of Southwest University of Science and Technology, 2010, 25(3): 22-24.
[7] Duan C Y, Hou X G, Zhang Z P, et al . Study on gardens utilizes of Viola in China. Chinese Agricultural Science Bulletin, 2004, 20(5): 185-186.
[8] Cai D F, Gao Z S, Si F H. Isozyme analyse of two cover plants Viola philippica and Viola prionantha . Grassland of China, 1998, 4(1): 48-49, 53.
[9] Wang X H, Qin M J, Yu G D. Viola medicinal plant: general survey and perspectives of its natural resources for exploitation. Chinese Wild Plant Resources, 2003, 22(4): 36-37.
[10] Ngan F, Chang R S, Tabba H D, et al . Isolation, purification and partial characterization of an active anti-HIV compound from the Chinese medical herb Viola yedoensis . Antiviral Research, 1988, 10(1-3): 107-115.
[11] Hildebrandt U, Hoef-Emden K, Backhausen S, et al . The rare, endemic zinc violets of Central Europe originate from Viola lutea Huds. Plant Systematics and Evolution, 2006, 257(3-4): 205-222.
[12] Noret N, Josens G, Escarré J, et al . Development of Issoria lathonia (Lepidoptera: Nymphalidae) on zinc-accumulating and nonaccumulating Viola species (Violaceae). Environmental Toxicology and Chemistry, 2007, 26(3): 565-571.
[13] Slomka A, Libik-Konieczny M, Kuta E, et al . Metalliferous and non-metalliferous populations of Viola tricolor represent similar mode of antioxidative response. Journal of Plant Physiology, 2008, 165(15): 1610-1619.
[14] Vengadesan G, Ganapathi A, Prem Anand R, et al . In vitro propagation of Acacia sinuate (Lour.) Merr. via cotyledonary nodes. Agroforestry Systems, 2002, 55: 9-15.
[15] Murashige T, Skoog F. Revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum, 1962, 15(3): 473-497.
[16] Wang X F, Wang C M, Zhang J L, et al . Development of a plant regeneration system via tissue culture in Puccinellia tenuiflora . Acta Prataculturae Sinica, 2014, 23(6): 355-360.
[17] Wang G, Du J, Li G Y, et al . Study on tissue cultural and clonal propagation of Lilium davidii var. Unicolor (Hoog) Cotton and Lilium brownie F. E. Brown ex Miellez. Journal of Northwest University (Natural Science), 2002, 38(1): 69-71.
[18] Meng Y L, Hao Y P, Jia J F, et al . Tissue cultural in vitro and plant regeneration from callus as well as explant of red clover ( Trifolium pratense L.). Acta Prataculturae Sinica, 1994, 3(2): 51-54.
[19] Du X H, Sun X L, Hao G P, et al . Effects of different explants and exogenous hormones on callus induction and growth of Ginkgo biloba in vitro . Journal of Shandong Normal University (Natural Science), 2008, 23(1): 129-133.
[20] Tan W C, Dai C G. Tissue Culture Technology of Ornamental[M]. Beijing: China Forestry Publishing House, 1991: 91-99.
[21] Zou Y M, Huang X F. Rapid micro-propagation and inflorescence induction of Tagetes erecta L. Journal of Jiangsu Forestry Science & Technology, 2005, 32(6): 17-19.
[22] Qi G X, Yang X D, Sui L, et al . Optimization of regeneration system of Trifolium repens L. Journal of Jilin Agricultural Sciences, 2011, 36(4): 51-54.
[23] Wu L J. Application and limitations of the woody plant tissue culture technique in the forestry scientific research and production. Journal of Fujian Forestry Science and Technology, 2003, 30(1): 67-70.
[24] Li C Y, Yin M H, Xu W H, et al . Effect of inorganic salt level and inositol concentration in MS medium and pH value on Phaseolus radiatus L. seed germination in test tube. Journal of Shangrao Normal College, 2008, 28(6): 62-65.
[25] Maene L, Debergh P. Liquid medium additions to established tissue cultures to improve elongation and rooting in vitro . Plant Cell Tissue & Organ Culture, 1984, 5(1): 23-33.
[26] López-Bucio J, Cruz-Ramírez A, Herrera-Estrella L. The role of nutrient availability in regulating root architecture. Current Opinion in Plant Biology, 2003, 6(3): 280-287.
[27] Dhiman M, Shatma V, Moitra S. Somatic embryogenesis and plant regeneration in Ephedra foliates (Boiss.); a non coniferous gymnosperm. Plant Tissue Culture and Biotechnology, 2010, 20(2): 133-143
[28] Qiao Q, Kong X S, Wu L, et al . Tissue culture and plant regeneration of Viola yedoensis . China Seed Industry, 2007, 6: 38-39.
[29] Chen Y Q, Liu J F, Zhong X, et al . Tissue culture and regeneration system from leaves of Saintpaulia ionantha in vitro . Chinese Agricultural Science Bulletin, 2010, 26(10): 212-216.
[30] Xu Q, Le Z B, Xu Y, et al . Study of leaf and petiole tissue culture induction and plant regeneration on sweet potato. Chinese Agricultural Science Bulletin, 2011, 27(15): 102-105.
[31] Zou Y C, Qin D J, Shuai C Q, et al . Study on tissue culture technique for rapid propagation of Cyclamen persicum . Journal of Hubei University for Nationalities (Natural Science Edition), 2011, 29(1): 23-27.
[1] 王丽娜. 早开堇菜( Viola prionantha Bunge)生活史及其生态适应性的研究[D]. 呼和浩特: 内蒙古大学, 2009.
[2] 刘会超, 孙振元.三种野生堇菜属植物引种及其园林应用. 中国园林, 2006, 1(13): 1-4.
[3] 张金政, 孙国峰, 刘雪川. 北京城市大环境绿化中可利用的草本花卉资源. 北京奥运和城市园林绿化建设论文集[C]. 北京: 北京园林学会, 2002.
[4] 周驰.紫花地丁的本草考证及化学成分研究[D]. 北京: 北京中医药大学, 2008: 13-15.
[5] 刘湘新, 刘进辉, 刘自逵, 等. 紫花地丁的有效成分分析及抗菌作用研究. 中兽医医药杂志, 2004, 23(3): 16-18.
[6] 陈红英. 早开堇菜的挥发性成分分析. 西南科技大学学报, 2010, 25(3): 22-24.
[7] 段春燕, 侯小改, 张赞平, 等. 堇菜属 Viola 植物资源的园林开发前景探讨.中国农学通报, 2004, 20(5): 185-186.
[8] 蔡朵芬, 高振声, 石飞华. 两种草坪地被植物紫花地丁与早开堇菜的同工酶研究.中国草地, 1998, 4(1): 48-49, 53.
[9] 王旭红, 秦民坚, 余国奠. 堇菜属药用植物研究概况与其资源利用前景.中国野生植物资源, 2003, 22(4): 36-37.
[16] 王雪芳, 王春梅, 张金林, 等. 小花碱茅组织培养植株再生体系的建立. 草业学报, 2014, 23(6): 355-360.
[17] 王刚, 杜捷, 李桂英, 等. 兰州百合和野百合组织培养及快速繁殖研究. 西北师范大学学报(自然科学版), 2002, 38(1): 69-71.
[18] 孟玉玲, 皓云鹏, 贾敬芬, 等. 红三叶组织培养及再生植株. 草业学报, 1994, 3(2): 51-54.
[19] 杜希华, 孙秀玲, 郝岗平. 不同外植体和激素对银杏愈伤组织诱导和生长的影响. 山东师范大学学报(自然科学版), 2008, 23(1): 129-133.
[20] 谭文澄, 戴策刚.观赏植物组织培养技术[M]. 北京:中国林业出版社, 1991: 91-99.
[21] 邹永梅, 黄雪芳. 万寿菊的组织培养和瓶苗开花研究. 江苏林业科技, 2005, 32(6): 17-19.
[22] 齐广勋, 杨向东, 隋丽, 等. 白三叶高频组织培养再生体系的研究. 吉林农业科学, 2011, 36(4): 51-54.
[23] 吴丽君. 木本植物组织培养技术在林业科研生产中的应用与局限. 福建林业科技, 2003, 30(1): 67-70.
[24] 李粹钰, 尹明华, 徐卫红, 等. MS培养基无机盐水平、肌醇浓度和pH值对绿豆试管内萌发的影响.上饶师范学院学报. 2008, 28(6): 62-65.
[28] 乔琦, 孔祥生, 伍丽, 等. 紫花地丁的组织培养和植株再生. 中国种业, 2007, 6: 38-39.
[29] 程云清, 刘剑锋, 钟雪, 等. 非洲紫罗兰叶片外植体再生技术体系的建立. 中国农学通报, 2010, 26(10): 212-216.
[30] 徐茜, 乐正碧, 徐燕, 等. 甘薯叶片和叶柄组织诱导培养及植株再生研究. 中国农学通报, 2011, 27(15): 102-105.
[31] 邹迎春, 覃大吉, 帅超群, 等. 仙客来组织培养快繁技术研究. 湖北民族学院学报(自然科学版), 2011, 29(1): 23-27.