4个鲜食葡萄品种组培快繁体系的建立
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摘要
为加快鲜食葡萄苗木的繁殖速度,提高苗木品质,以鲜食葡萄品种夏黑、红地球、巨峰和玫瑰香为试验材料,通过对无菌外植体的消毒、继代和驯化等环节技术的探究,建立适用于不同鲜食葡萄品种的组培快繁体系。结果表明,以葡萄茎段作为外植体在低浓度(0.3%～0.5%)次氯酸钠溶液中消毒15 min后,接种在MS+0.2 mg·L~(-1)IBA+1.0 mg·L~(-1)6-BA+0.5 mg·L~(-1)KT+4.0 mg·L~(-1)腺嘌呤+30 g·L~(-1)蔗糖+8.2 g·L~(-1)琼脂的培养基上,成活率达到87.5%;以培养基WPM添加0.2 mg·L~(-1 )IBA增殖培养兼生根诱导,组培苗生长健壮,生根效果好。采用水培方法炼苗3周后4个品种组培苗移栽成活率均达到100%。综上,完整的基础组培快繁体系可应用于科学研究及工业化大规模生产。本研究为加快鲜食葡萄的繁殖速度、提高苗木质量提供了理论依据。
In order to speed up the propagation and improve the quality of nursery plants of table grapes, in vitro mircropropagation was established. In this study, cultivars of Summer Black, Red Globe, Kyoho and Muscat Hamburg were used as materials to explore the methods of disinfection of aseptic explants, the composition of subculture medium and the techniques of domestication. The results showed that 87.6% of explants of nodal stems survived when sterilized by the low concentration of 0.3%～0.5% sodium hypochlorite for 15 min and cultured on the MS medium supplementing with 0.2 mg·L~(-1) IBA, 1.0 mg·L~(-1) 6-BA and 0.5 mg·L~(-1) KT. The propagules grew robustly on the WPM medium supplementing with IBA 0.2 mg·L~(-1) that was suitable for both shoot growth and rooting, and the proliferation rate of shoots were enhanced. All plantlets of four table grape cultivars survived when transplanted in the green house after hydroponic acclimation for 3 to 4 weeks. These results suggested that this in-vitro rapid propagating system was basically suitable for four table grape cultivars and could be applied to scientific research and industrialized mass production of nursery plants. This study provides theoretical basis for accelerating the propagation speed of fresh grape and improving seedling quality.
In order to speed up the propagation and improve the quality of nursery plants of table grapes, in vitro mircropropagation was established. In this study, cultivars of Summer Black, Red Globe, Kyoho and Muscat Hamburg were used as materials to explore the methods of disinfection of aseptic explants, the composition of subculture medium and the techniques of domestication. The results showed that 87.6% of explants of nodal stems survived when sterilized by the low concentration of 0.3%～0.5% sodium hypochlorite for 15 min and cultured on the MS medium supplementing with 0.2 mg·L~(-1) IBA, 1.0 mg·L~(-1) 6-BA and 0.5 mg·L~(-1) KT. The propagules grew robustly on the WPM medium supplementing with IBA 0.2 mg·L~(-1) that was suitable for both shoot growth and rooting, and the proliferation rate of shoots were enhanced. All plantlets of four table grape cultivars survived when transplanted in the green house after hydroponic acclimation for 3 to 4 weeks. These results suggested that this in-vitro rapid propagating system was basically suitable for four table grape cultivars and could be applied to scientific research and industrialized mass production of nursery plants. This study provides theoretical basis for accelerating the propagation speed of fresh grape and improving seedling quality.
引文
[1] 刘凤之. 中国葡萄栽培现状与发展趋势[J]. 落叶果树, 2017,49 (1): 1-4
[2] 晁无疾. 中国葡萄品牌建设现状及发展展望[J]. 中外葡萄与葡萄酒, 2016(5):135-140
[3] 王秀芬, 刘俊, 李敬川, 于祎飞, 刘寅喆. 转型升级是中国葡萄产业可持续发展的必由之路[J]. 中外葡萄与葡萄酒, 2017(4):117-123
[4] Heloir M C, Fournioux J C, Oziol L, Bessis R. An improved procedure for the propagation in vitro of grapevine (Vitis vinifera cv. Pinot noir) using axillary bud microcuttings[J]. Plant Cell Tissue and Organ Culture, 1997, 49: 223-225
[5] 闵首军, 李爱玲. 葡萄组织快繁技术研究[J]. 农学学报, 2012, 2(10):55-57
[6] 陈斌. 葡萄茎尖培养脱毒与快繁技术研究[D]. 长沙: 湖南农业大学, 2015
[7] Chuong P Ⅴ, Beversdorf W D. High frequency embry ogenesis through isolated microspore culture in Brassic a napus L. and B. carinata braun[J]. Plant Science, 1985, 39(3): 219-226
[8] Gray D J, Benton C M. Micropropagation and plant establishment of muscadine grape[J]. Proceedings of the Florida State Horticultural Society, 1990, 103: 300-302
[9] Gray D J, Benton C M. In vitro micropropagation and plant establishment of muscadine grape cultivars (Vitis rotundifolia)[J]. Plant Cell Tissue and Organ Culture, 1991, 27: 7-14
[10] 沈传进, 王利民, 张平, 闫云花, 安旭军, 李振德. 鲜食葡萄组织培养快速繁育系统的建立[J]. 河北农业科学, 2011,15(7):16-21
[11] 曾建国, 吴雪梅. 巨峰葡萄组培快繁过程中初代培养基的筛选[J]. 现代园艺, 2010(7):4-7
[12] 刘娜, 许轲, 张文, 王琢, 朱元娣. 四个酿酒葡萄品种组培快繁体系的初建[J]. 植物生理学报, 2013, 49(10): 1071-1076
[13] Chee R, Pool R M. The effects of growth substances and photoperiod on the development of shoot apices of Vitis cultured in vitro[J]. Scientia Horticultureae, 1982, 16: 17-27
[14] 黄丽燕, 张雅丽, 卢江. 葡萄砧木品种离体快繁研究初报[J]. 中外葡萄与葡萄酒, 2010(5):14-17
[15] Diab A A, Khalil S M, Ismail R M. Regeneration and micropropagation of grapevine (Vitis vinifera) through shoot tips and axillary buds[J]. International Journal of Advanced Biotechnology Research, 2011, 2(4): 484-491
[16] Jaskani M J, Haider A, Sultana R, Khan M M, Qasim M, Iqrar AK. Effect of growth hormones on micropropagation of Vitis vinifera L. cv. Perlette[J]. Pakistan Journal of Botany, 2008, 40 (1): 105-109
[17] 潘学军, 张文娥, 唐冬梅, 骆强伟. 无核葡萄离体快繁技术研究[J]. 中国南方果树, 2007,36(2): 44-46
[18] 王鹏飞, 杜俊杰. 果树试管苗的炼苗移栽技术[J]. 山西果树, 2004,3:20-22
[19] Arnon D Ⅰ, Hoagland D R. The investigation of plant nutrition by artificial culture methods[J]. Biological Reviews, 1994, 19 (2):55-67
[20] 谢璇, 许轲, 谢闽新, 朱元娣. 苹果茎尖培养快繁体系的优化[J]. 植物生理学报, 2015, 51(12): 2152-2156
[21] 冯文华, 代红军.‘赤霞珠’葡萄组培快繁体系研究[J]. 北方园艺, 2016(13): 104-106
[22] 李国树, 范树国, 邱璐, 徐成东, 王波, 李天星,李军, 李亚东. 水晶葡萄茎尖组织研究[J]. 中国南方果树, 2010, 39(2): 66-67
[23] 周恒, 万召娣, 罗静. 紫秋刺葡萄离体繁殖研究初报[J]. 贵州农业科学, 2010, 38(9): 22-25
[24] 王冬梅, 黄学林, 黄上志. 细胞分裂素类物质在植物组织培养中的作用机制[J]. 植物生理学通讯, 1996, 32(5):373-377
[25] 洪森荣, 尹明华. 香果树带芽茎段不定芽高频增殖的优化[J]. 核农学报, 2010, 24(3): 532-536
[26] 邓年方, 王义强. 植物组织培养快速繁殖类型综述[J]. 江苏林业科技, 2006, 33(1): 41-44
[27] 冯发文, 田群芳. 巨峰葡萄组织培养快繁技术研究[J]. 落叶果树, 2011(2): 6-8
[28] 曹孜义, 杨德龙, 梁庆丰, 于作文. 内39号葡萄株系的离体快繁技术研究[J]. 果树学报, 2002, 19(6): 427-429
[29] 韩艳婷. 四种葡萄组织培养体系的构建及卷叶病毒脱除与检测研究[D]. 长沙: 湖南农业大学,2008
[2] 晁无疾. 中国葡萄品牌建设现状及发展展望[J]. 中外葡萄与葡萄酒, 2016(5):135-140
[3] 王秀芬, 刘俊, 李敬川, 于祎飞, 刘寅喆. 转型升级是中国葡萄产业可持续发展的必由之路[J]. 中外葡萄与葡萄酒, 2017(4):117-123
[4] Heloir M C, Fournioux J C, Oziol L, Bessis R. An improved procedure for the propagation in vitro of grapevine (Vitis vinifera cv. Pinot noir) using axillary bud microcuttings[J]. Plant Cell Tissue and Organ Culture, 1997, 49: 223-225
[5] 闵首军, 李爱玲. 葡萄组织快繁技术研究[J]. 农学学报, 2012, 2(10):55-57
[6] 陈斌. 葡萄茎尖培养脱毒与快繁技术研究[D]. 长沙: 湖南农业大学, 2015
[7] Chuong P Ⅴ, Beversdorf W D. High frequency embry ogenesis through isolated microspore culture in Brassic a napus L. and B. carinata braun[J]. Plant Science, 1985, 39(3): 219-226
[8] Gray D J, Benton C M. Micropropagation and plant establishment of muscadine grape[J]. Proceedings of the Florida State Horticultural Society, 1990, 103: 300-302
[9] Gray D J, Benton C M. In vitro micropropagation and plant establishment of muscadine grape cultivars (Vitis rotundifolia)[J]. Plant Cell Tissue and Organ Culture, 1991, 27: 7-14
[10] 沈传进, 王利民, 张平, 闫云花, 安旭军, 李振德. 鲜食葡萄组织培养快速繁育系统的建立[J]. 河北农业科学, 2011,15(7):16-21
[11] 曾建国, 吴雪梅. 巨峰葡萄组培快繁过程中初代培养基的筛选[J]. 现代园艺, 2010(7):4-7
[12] 刘娜, 许轲, 张文, 王琢, 朱元娣. 四个酿酒葡萄品种组培快繁体系的初建[J]. 植物生理学报, 2013, 49(10): 1071-1076
[13] Chee R, Pool R M. The effects of growth substances and photoperiod on the development of shoot apices of Vitis cultured in vitro[J]. Scientia Horticultureae, 1982, 16: 17-27
[14] 黄丽燕, 张雅丽, 卢江. 葡萄砧木品种离体快繁研究初报[J]. 中外葡萄与葡萄酒, 2010(5):14-17
[15] Diab A A, Khalil S M, Ismail R M. Regeneration and micropropagation of grapevine (Vitis vinifera) through shoot tips and axillary buds[J]. International Journal of Advanced Biotechnology Research, 2011, 2(4): 484-491
[16] Jaskani M J, Haider A, Sultana R, Khan M M, Qasim M, Iqrar AK. Effect of growth hormones on micropropagation of Vitis vinifera L. cv. Perlette[J]. Pakistan Journal of Botany, 2008, 40 (1): 105-109
[17] 潘学军, 张文娥, 唐冬梅, 骆强伟. 无核葡萄离体快繁技术研究[J]. 中国南方果树, 2007,36(2): 44-46
[18] 王鹏飞, 杜俊杰. 果树试管苗的炼苗移栽技术[J]. 山西果树, 2004,3:20-22
[19] Arnon D Ⅰ, Hoagland D R. The investigation of plant nutrition by artificial culture methods[J]. Biological Reviews, 1994, 19 (2):55-67
[20] 谢璇, 许轲, 谢闽新, 朱元娣. 苹果茎尖培养快繁体系的优化[J]. 植物生理学报, 2015, 51(12): 2152-2156
[21] 冯文华, 代红军.‘赤霞珠’葡萄组培快繁体系研究[J]. 北方园艺, 2016(13): 104-106
[22] 李国树, 范树国, 邱璐, 徐成东, 王波, 李天星,李军, 李亚东. 水晶葡萄茎尖组织研究[J]. 中国南方果树, 2010, 39(2): 66-67
[23] 周恒, 万召娣, 罗静. 紫秋刺葡萄离体繁殖研究初报[J]. 贵州农业科学, 2010, 38(9): 22-25
[24] 王冬梅, 黄学林, 黄上志. 细胞分裂素类物质在植物组织培养中的作用机制[J]. 植物生理学通讯, 1996, 32(5):373-377
[25] 洪森荣, 尹明华. 香果树带芽茎段不定芽高频增殖的优化[J]. 核农学报, 2010, 24(3): 532-536
[26] 邓年方, 王义强. 植物组织培养快速繁殖类型综述[J]. 江苏林业科技, 2006, 33(1): 41-44
[27] 冯发文, 田群芳. 巨峰葡萄组织培养快繁技术研究[J]. 落叶果树, 2011(2): 6-8
[28] 曹孜义, 杨德龙, 梁庆丰, 于作文. 内39号葡萄株系的离体快繁技术研究[J]. 果树学报, 2002, 19(6): 427-429
[29] 韩艳婷. 四种葡萄组织培养体系的构建及卷叶病毒脱除与检测研究[D]. 长沙: 湖南农业大学,2008