[an error occurred while processing this directive] [an error occurred while processing this directive] [an error occurred while processing this directive]
[an error occurred while processing this directive]
技术方法

栽培薯蓣茎段离体再生体系的建立

  • 冯雯 ,
  • 王玉国
展开
  • 复旦大学生命科学学院, 上海 200438
*王玉国, 复旦大学生命科学学院教授, 博士生导师。长期从事植物系统发育基因组学、进化生态学和重要遗传资源保护与利用研究。主持多项国家自然科学基金和省部级科研项目; 在Journal of Experimental Botany、Molecular Ecology Resources、PLoS Genetics、Molecular Phylogenetics and Evolution及生物多样性等国内外重要学术期刊上发表研究论文80余篇。目前其研究团队主要采用系统学、基因组学、进化生态学和生物信息学等多学科手段, 结合经典研究方法, 探讨复杂植物类群的种间关系与适应性进化机制, 开展重要作物的起源与进化和特有种质资源评价、保护与创新研究。E-mail: wangyg@fudan.edu.cn

收稿日期: 2024-04-08

  录用日期: 2024-05-27

  网络出版日期: 2024-05-27

基金资助

上海市崇明区农业科创项目(2021CNKC-01-02)

Establishment of an In Vitro Regeneration System for Stem Segments of Cultivated Dioscorea polystachya

  • Wen Feng ,
  • Yuguo Wang
Expand
  • School of Life Sciences, Fudan University, Shanghai 200438, China

Received date: 2024-04-08

  Accepted date: 2024-05-27

  Online published: 2024-05-27

摘要

为建立栽培薯蓣(Dioscorea polystachya)组培再生体系, 以茎尖及茎尖下4个不同长度的茎段为外植体, 探讨了不同浓度植物生长调节剂、培养基类型及茎段对腋芽诱导和植株再生的影响。结果表明, 培养基配比为MS+1.0 mg∙L-1 6-BA+0.5 mg∙L-1 KT和茎尖下12-20 cm的茎段为栽培薯蓣不定芽诱导的最佳组合, 诱导率达90.0%; 继代培养最适培养基配方为MS+0.5 mg∙L-1 6-BA+0.05 mg∙L-1 NAA+0.1 mg∙L-1 KT, 低浓度NAA固定时, 增殖系数在一定范围内随着6-BA浓度的升高而增高; 在相同外源激素条件下, 快繁生根采用DKW培养基明显优于常规1/2MS培养基, 生根率显著提高, 达92.86%。添加1 mg PVP以及增加转接次数均能明显降低褐化率。该研究有效解决了栽培薯蓣种质资源离体稳定繁殖取材部位优化的问题, 为规模化生产栽培薯蓣优质脱毒苗奠定了良好基础。

本文引用格式

冯雯 , 王玉国 . 栽培薯蓣茎段离体再生体系的建立[J]. 植物学报, 2024 , 59(5) : 792 -799 . DOI: 10.11983/CBB24051

Abstract

To establish a tissue culture regeneration system for cultivated Chinese yam (Dioscorea polystachya), the effects of different concentrations of plant growth regulators, and stem segments on axillary bud induction and plant regeneration were investigated with different concentrations of plant growth regulators, media types, and shoot tips and stem segments at four different positions under the shoot tip as explants. The results revealed that MS+1.0 mg∙L-1 6-BA+0.5 mg∙L-1 KT hormone ratio and 12-20 cm stem segment under the stem tip were the best formulas for the induction of adventitious buds in D. polystachya, and the induction rate reached 90.0%. In the subculture, the optimal concentration was MS+0.5 mg∙L-1 6-BA+0.05 mg∙L-1 NAA+0.1 mg∙L-1 KT, and the proliferation coefficient increased with increasing of 6-BA within a certain range when the low concentration of NAA was constant. With the same concentration of exogenous hormones, DKW medium was significantly better than the conventional 1/2MS medium, and the rooting rate is significantly improved, reaching 92.86%. Adding 1 mg PVP (polyvinylpyrrolidone) and increasing the number of transfers can significantly reduce browning. This study effectively solved the problem of optimizing the explant position for the stable of in vitro propagation of cultivated yam germplasm resources, laying a good foundation for the large-scale production of their high-quality virus-free seedlings.

[an error occurred while processing this directive]

参考文献

[1] Ammirato PV ( Crop Species. 1984). Yams. In: Ammirato PV, Evans DA, Sharp WR, Yamada Y, eds. Handbook of Plant Cell Culture, Vol. 3. New York: Macmillan. pp. 327- 354.
[2] Anike FN, Konan K, Olivier K, Dodo H (2012). Efficient shoot organogenesis in petioles of yam (Dioscorea spp). Plant Cell Tissue Organ Cult 111, 303-313.
[3] Baker R, Phillips DJ (1962). Obtaining pathogen-free stock by shoot tip culture. Phytopathology 52, 1242-1244.
[4] Blunden G, Hardman R, Trease GE (1966). Some observations on the propagation of Dioscorea belizensis Lundell and other steroid-yielding yams. Planta Med 14, 84- 89.
[5] Cabanillas E, Martin FW (1978). The propagation of edible yams from cuttings. J Agric Univ Puerto Rico 62, 249- 254.
[6] Chen YQ, Fan JY, Yi F, Luo ZX, Fu YS (2003). Rapid clonal propagation of Dioscorea zingiberensis. Plant Cell Tissue Organ Cult 73, 75-80.
[7] Chen ZH, Hua SM, Li LH, Luo XF, He PZ (2018). Optimization of technical conditions for tissue culture and rapid propagation of Chinese yam. Fujian J Agric Sci 33, 164- 170. (in Chinese)
  陈芝华, 华树妹, 李丽红, 罗晓锋, 贺佩珍 (2018). 山药组培快繁技术条件的优化. 福建农业学报 33, 164-170.
[8] Hang YY, Qin HZ, Ding ZZ (1992). A survey and quality research on the new resources of Shan Yao (rhizoma dioscoreae). J Plant Resour Environ 1(2), 10-15. (in Chinese)
  杭悦宇, 秦慧贞, 丁志遵 (1992). 山药新药源的调查和质量研究. 植物资源与环境 1(2), 10-15.
[9] He Z, Huang T, Qin SL, Cui CY, Jiang JX (2016). Pathogen identification of Ruichang yam virus disease. In: Chinese Society for Plant Pathology (CSPP), ed. Proceedings of the 2016 Academic Annual Meeting of CSPP. Beijing: China Agricultural Science and Technology Press. pp. 305. (in Chinese)
  贺哲, 黄婷, 秦双林, 崔朝宇, 蒋军喜 (2016). 瑞昌山药病毒病病原鉴定. 见:中国植物病理学会. 中国植物病理学会2016年学术年会论文集. 北京: 中国农业科学技术出版社. pp. 305.
[10] Hu WB, Zhang SF, Sun N (2018). Stem tissue culture of red grape in Longnan area. Shaanxi Forest Sci Technol 46, 6-9. (in Chinese)
  胡文斌, 张少飞, 孙娜 (2018). 陇南红提葡萄茎段组织培养研究. 陕西林业科技 46, 6-9.
[11] Ishizaki T (2021). A tissue culture system for callus formation and plant regeneration using tuber discs of Dioscorea tokoro Makino, a pharmaceutical yam species. Biologia 76, 1141-1145.
[12] Kohmura H, Araki H, Imoto M (1995). Micropropagation of ‘Yamatoimo’ Chinese yam (Dioscorea opposita) from immature leaves. Plant Cell Tissue Organ Cult 40, 271-276.
[13] Li MJ, Guo J, Li X, Li JQ, Wang YP, Zhang XL, Liu YK (2012). A regeneration system and ploidy identification of plantlets for endosperm of Dioscorea zingiberensis. Chin Bull Bot 47, 654-660. (in Chinese)
  李明军, 郭婧, 李翔, 李纪强, 王医鹏, 张晓丽, 刘永康 (2012). 盾叶薯蓣胚乳再生体系的建立及其染色体倍性鉴定. 植物学报 47, 654-660.
[14] Li MJ, Zhang JB, Zhang HB (2000). Study on the callus induction and plantlet regeneration from the bulbil of Dioscorea opposita. Acta Bot Boreal-Occident Sin 20, 772- 777. (in Chinese)
  李明军, 张嘉宝, 张海波 (2000). 怀山药零余子愈伤组织诱导及植株再生的研究. 西北植物学报 20, 772-777.
[15] Li Q, Yang B, Li HL, Fu ZH (2004). Research on tissue rapid propagation of a specified cultivar “Foshou” yam from Hubei province. In: Chinese Society for Agricultural Biotechnology, Fujian Agriculture and Forestry University eds. Proceedings of the 2nd National Symposium on Plant Tissue Culture, Detoxification and Rapid Propagation, and Industrial Production. Beijing: China Agricultural Science and Technology Press. pp. 96-101. (in Chinese)
  李琼, 杨波, 李洪林, 付志惠 (2004). 一种特有经济作物“佛手山药”的组织快繁研究. 见:中国农业生物技术学会, 福建农林大学. 第二届全国植物组织培养、脱毒快繁及工厂化生产学术研讨会论文集. 北京: 中国农业科学技术出版社. pp. 96-101.
[16] Li RZ (2005). Method for tissue culture of Huaishan yam. Chinese patent, ZL1653888A. 2005-08-17. (in Chinese)
  李锐忠 (2005). 淮山(山药)组织培养方法. 中国专利, ZL1653888A. 2005-08-17.
[17] Li Y, Ji SY, Xu T, Zhong YH, Xu MH, Liu YQ, Li MJ, Fan B, Wang FZ, Xiao JB, Lu BY (2023). Chinese yam (Dioscorea): nutritional value, beneficial effects, and food and pharmaceutical applications. Trends Food Sci Technol 134, 29-40.
[18] Liang FG, Ma KL, Li MJ, Liu AX (2001). Effect of different explant of Dioscrea opposita on callus induction and plantlets regeneration. J Henan Vocation-Tech Teachers College 29, 24-27. (in Chinese)
  梁方刚, 马克莉, 李明军, 刘爱霞 (2001). 怀山药不同外植体对愈伤组织形成和植株再生的影响. 河南职技师院学报 29, 24-27.
[19] Liu JY, Xu YM, Li SL, Guo CF, Guo ZC (2006). Tissue culture of Dioscorea opposita “foshou”. Bull Bot Res 26, 323-328. (in Chinese)
  刘金英, 徐有明, 李双来, 郭春凤, 郭治成 (2006). 佛手山药组织培养的研究. 植物研究 26, 323-328.
[20] Liu M, Jiang FL, Kong XY, Tian J, Wu ZX, Wu Z (2017). Effects of multiple factors on hyperhydricity of Allium sativum L. Sci Hortic 217, 285-296.
[21] Liu YN, Chen YL, He XB, Hong X, Mi M (2022). Tissue culture technology of Chinese yam. Bot Res 11, 560-563. (in Chinese)
  刘也楠, 陈银龙, 何贤彪, 洪霞, 米敏 (2022). 山药的组织培养技术. 植物学研究 11, 560-563.
[22] Liu ZH, Dai XH, Li J, Liu N, Liu XZ, Li S, Xiang FN (2020). The type-B cytokinin response regulator ARR1 inhibits shoot regeneration in an ARR12-dependent manner in Arabidopsis. Plant Cell 32, 2271-2291.
[23] Long Y, Yang Y, Pan GT, Shen YO (2022). New insights into tissue culture plant-regeneration mechanisms. Front Plant Sci 13, 926752.
[24] Matosevich R, Cohen I, Gil-Yarom N, Modrego A, Friedlander-Shani L, Verna C, Scarpella E, Efroni I (2020). Local auxin biosynthesis is required for root regeneration after wounding. Nat Plants 6, 1020-1030.
[25] Nagasawa A, Finer JJ (1989). Plant regeneration from embryogenic suspension cultures of Chinese yam (Dioscorea opposita Thunb.). Plant Sci 60, 263-271.
[26] Nan HL, Liu JP, Wang YQ (2005). Research on the propagation technology of yam stem tip. Crops (6), 34-36. (in Chinese)
  南怀林, 刘建平, 王耀琴 (2005). 山药茎尖繁殖技术的研究. 作物杂志 (6), 34-36.
[27] Ng SYC (1988). In vitro tuberization in white yam (Dioscorea rotundata Poir). Plant Cell Tissue Organ Cult 14, 121- 128.
[28] Ng SYC, Hahn SK (1985). Application of tissue culture to tuber crops at IITA. In: Proceedings of the Inter-Center Seminar on International Agricultural Research Centers (IARCs) and Biotechnology. Los Baños: IRRI. pp. 29-40.
[29] Pan M, Huang S, Wang JF, Lü DR, Qi HS, Jiang DQ (2014). Study on in-vitro cultivation conditions and cultivating techniques of Dioscorea opposite. Guizhou Agric Sci 42(3), 125-129. (in Chinese)
  潘梅, 黄赛, 王景飞, 吕德任, 戚华莎, 姜殿强 (2014). 山药茎段的离体培养与育苗基质筛选. 贵州农业科学 42(3), 125-129.
[30] Ping AM, Hou LP, Xing GM, Li ML (2014). Study on tissue culture from shoot tip and stem segment of Dioscorea opposite Thunb. cv. ‘Tiegun’. Heilongjiang Agric Sci (9), 6-10. (in Chinese)
  平阿敏, 侯雷平, 邢国明, 李梅兰 (2016). 铁棍山药茎尖组培及茎段侧芽成苗研究. 黑龙江农业科学 (9), 6-10.
[31] Skoog F, Miller CO (1957). Chemical regulation of growth and organ formation in plant tissues cultured in vitro. Symp Soc Exp Biol 11, 118-130.
[32] Terry ER, Hahn SK (1980). The effect of cassava mosaic disease on growth and yield of a local and an improved variety of cassava. Trop Pest Manage 26, 34-37.
[33] Thresh JM, Fargette D, Otim-Nape GW (1994). Effects of African cassava mosaic geminivirus on the yield of cassava. Trop Sci 34, 26-42.
[34] Twyford CT, Mantell SH (1996). Production of somatic embryos and plantlets from root cells of the Greater Yam. Plant Cell Tissue Organ Cult 46, 17-26.
[35] Vander Zaag P, Fox RL (1981). Field production of yams (Dioscorea alata) from stem cuttings. Trop Agric 58, 143- 145.
[36] Viana AM, Mantell SH (1989). Callus induction and plant regeneration from excised zygotic embryos of the seed-propagated yams Dioscorea composita Hemsl. and D. cayenensis Lam. Plant Cell Tissue Organ Cult 16, 113- 122.
[37] Wang HJ, Wang TL, Bai ZW, Xu YX, Li MJ (2006). The effect of hormone ratio on inducing adventitious buds from different explants of Chinese yam. Henan Agric Sci 35 (12), 73-74. (in Chinese)
  王红娟, 王天亮, 白自伟, 徐艳霞, 李明军 (2006). 激素配比对怀山药不同外植体诱导不定芽的影响. 河南农业科学 35(12), 73-74.
[38] Zhu YF, Yin HQ, Liu HM, Wang YL, Xiang JQ, Yang YK, Ran Q, Ye ZY, Cheng Q, Xu Y, Chen QL, Tian TF (2015). A rapid propagation method for tissue culture of Lichuan yam. Chinese patent, ZL105145373A. 2015-12- 16. (in Chinese)
  朱云芬, 殷红清, 刘河明, 王应玲, 向极钎, 杨永康, 冉强, 叶紫云, 程群, 徐怡, 陈巧玲, 田腾飞 (2015). 一种利川山药组培快繁方法. 中国专利, ZL105145373A. 2015-12-16.
文章导航

/

[an error occurred while processing this directive]