何首乌离体快繁技术体系的建立
|
摘要
【目的】建立何首乌离体快繁技术体系,为其种苗生产提供技术支持。【方法】以从广西陆川县采集的何首乌幼嫩单芽茎段为外植体,分析不同灭菌时间、不同植物生长调节剂种类和浓度及不同培养方式对其组织培养快繁技术的影响。【结果】适宜何首乌外植体消毒灭菌的方法为0.1%HgCl2浸泡处理8 min;适宜不定芽诱导的初代培养基为MS+1.0 mg/L 6-氨基腺嘌呤(6-BA)+0.05 mg/L a-萘乙酸(NAA),腋芽萌发率达96.7%;适宜的继代增殖培养基为MS+2.0 mg/L苯基噻二唑脲(TDZ)+0.05 mg/L NAA,增殖系数为5.68,植株健壮;无菌苗的单芽茎段以浸没间歇式培养(TIBs)增殖系统效果最佳;适宜的生根培养基为1/2MS+0.01 mg/L NAA,生根率达91.33%。【结论】建立了以带腋芽茎段为外植体的何首乌离体快繁技术体系,可为其种苗生产提供新途径。
【Objective】The present study was conducted to develop rapid propagation technology of Polygonum multiforum in vitro and provide reference for seeding-use seedling production.【Method】The single-bud stems of P.multiforum fromLuchuan,Guangxi were as the explants.The effects of different sterilization times,types and concentrations of plant growth regulators and different culture methods on the tissue culture rapid propagation technology of P.multiforum were explored.【Result】The results showed that,the best explant sterilization effects for explants was achieved by immersing the explants in 0.1% HgCl2 solution for 8 min.The suitable primary medium for adventitious bud induction was MS+1.0mg/L 6-aminadenine(6-BA)+0.05 mg/L a-naphthylacetic acid(NAA),and the axillary bud germination rate was 96.7%under this condition.The suitable subculture medium was MS+2.0 mg/L phenothiadiazolylurea(TDZ)+0.05 mg/L NAA,and the multiplication coefficient was 5.68.The plantlets were strong.Temporary immersion system(TIBs)culture was suitable for single bud stem segment of P.multiforum aseptic seedlings.The suitable medium for rooting was 1/2MS+0.01mg/L NAA,and the rooting rate was 91.33%.【Conclusion】The in vitro rapid propagation system for P.multiforum explants with axillary bud is established,it can provide new method for producing seed-using seedlings.
【Objective】The present study was conducted to develop rapid propagation technology of Polygonum multiforum in vitro and provide reference for seeding-use seedling production.【Method】The single-bud stems of P.multiforum fromLuchuan,Guangxi were as the explants.The effects of different sterilization times,types and concentrations of plant growth regulators and different culture methods on the tissue culture rapid propagation technology of P.multiforum were explored.【Result】The results showed that,the best explant sterilization effects for explants was achieved by immersing the explants in 0.1% HgCl2 solution for 8 min.The suitable primary medium for adventitious bud induction was MS+1.0mg/L 6-aminadenine(6-BA)+0.05 mg/L a-naphthylacetic acid(NAA),and the axillary bud germination rate was 96.7%under this condition.The suitable subculture medium was MS+2.0 mg/L phenothiadiazolylurea(TDZ)+0.05 mg/L NAA,and the multiplication coefficient was 5.68.The plantlets were strong.Temporary immersion system(TIBs)culture was suitable for single bud stem segment of P.multiforum aseptic seedlings.The suitable medium for rooting was 1/2MS+0.01mg/L NAA,and the rooting rate was 91.33%.【Conclusion】The in vitro rapid propagation system for P.multiforum explants with axillary bud is established,it can provide new method for producing seed-using seedlings.
引文
陈菊,陈国惠.2006a.何首乌生根培养因子的优选研究[J].西南农业大学学报(自然科学版),28(5):756-758.[Chen J,Chen G H.2006a.Optimization of the culture conditions for the rooting of Polygonum multiforum plantlets[J].Journal of Southwest Agricultural University(Natural Science),28(5):756-758.]
陈菊,陈国惠.2006b.6-BA与NAA浓度配比对何首乌不定芽增殖的影响[J].中国农学通报,22(11):173-175.[Chen J,Chen G H.2006b.Study of the concentration of 6-BAand NAA in the rapid propagation of Polygonum multiflorum Thunb[J].Chinese Agricultural Science Bulletin,22(11):173-175.]
高美萍,林志城,张驰,江文,董伟清,欧昆鹏,桂杰,闭志强,何芳练,陈丽娟.2016.荸荠组培苗在间歇浸没式生物反应器(TIBs)中的高效增殖技术[J].南方农业学报,47(10):1653-1657.[Gao M P,Lin Z C,Zhang C,Jiang W,Dong W Q,Ou K P,Gui J,Bi Z Q,He F L,Chen LJ.2016.Efficient propagation of Eleocharis dulcis plantlets in temporary immersion bioreactors(TIBs)[J].Journal of Southern Agriculture,47(10):1653-1657.]
贾梦雪,徐瑾,叶香娟,刘芊,王喆,刘燕.2013.春石斛优良品种‘森禾2006’组培快繁体系的建立[J].植物生理学报,49(12):1363-1367.[Jia M X,Xu J,Ye X J,Liu Q,Wang Z,Liu Y.2013.Establishment of rapid propagation system for elite variety of Dendrobium nobile Lindl.‘Senhe 2006’by tissue culture[J].Plant Physiology Journal,49(12):1363-1367.]
李晶,罗玉洁,张青林,罗正荣,刘继红.2016.君迁子休眠芽及叶片离体培养体系优化及植株再生[J].华中农业大学学报,35(4):14-19.[Li J,Luo Y J,Zhang Q L,Luo Z R,Liu J H.2016.In vitro culture system optimization and regeneration of date plum(Diospyros lotus Linn.)dormant buds and leaves[J].Journal of Huazhong Agricultural University,35(4):14-19.]
李娟玲,刘国民,邱文华,徐立新.2003.何首乌茎段离体培养的研究[J].贵州科学,21(3):86-91.[Li J L,Liu G M,Qiu W H,Xu L X.2003.Study on the stem segments culture in vitro of Polygonum multiflorum Thunb[J].Guizhou Science,21(3):86-91.]
刘义存,黄天启,林顺权.2017.枇杷属植物野生种的茎段愈伤组织诱导植株再生研究[J].果树学报,34(6):762-768.[Liu Y C,Huang T Q,Lin S Q.2017.Study on plant regeneration of callus from the stem of wild loquat[J].Journal of Fruit Science,34(6):762-768.]
龙滢,杨鑫,余春香.2005.何首乌的组织培养研究[J].中医药导报,11(11):63-64.[Long Y,Yang X,Yu C X.2005.Tissue culture research on fleece flower root[J].Guiding Journal of TCM,11(11):63-64.]
孙桂波,纪凤兰,徐惠波,孙晓波.2007.何首乌的化学成分与药理作用研究进展[J].长春中医药大学学报,23(4):105.[Sun G B,Ji F L,Xu H B,Sun X B.2007.Advances in studies on the chemical components and pharmacological activities of Radix polygoni multiflora[J].Journal of Changchun University of Traditional Chinese Medicine,23(4):105.]
孙英坤,胡绍庆,庞基良,高凯,刘华红,陈焕伟,姚涛,陈林敬,沈柏春.2017.珍稀濒危物种堇叶紫金牛高效快繁体系的建立[J].植物学报,52(6):764-773.[Sun Y K,Hu S Q,Pang J L,Gao K,Liu H H,Chen H W,Yao T,Chen L J,Shen B C.2017.Establishment of a tissue culture and propagation system for Ardisia violacea,a rare and endangered species[J].Chinese Bulletin of Botany,52(6):764-773.]
魏岳荣,杨护,黄秉智,黄霞,黄学林,邱继水,许林兵.2007.Picloram、ABA和TDZ对香蕉体细胞胚胎发生的影响[J].园艺学报,34(1):81-86.[Wei Y R,Yang H,Huang B Z,Huang X,Huang X L,Qiu J S,Xu L B.2007.Effects of Picloram,ABA and TDZ on somatic embryogenesis of banana[J].Acta Horticulturae Sinica,34(1):81-86.]
吴光洪,孙英坤,陈林敬,沈柏春,邓梦楚,钱飞.2016.兔眼蓝莓‘粉蓝’叶片直接诱导丛生芽再生技术体系的建立[J].植物生理学报,52(3):372-380.[Wu G H,Sun Y C,Chen L J,Shen B C,Deng M C,Qian F.2016.The technology system establishment for direct induction of Vaccinium ashei‘Powderblue’leaves to regenerate multiple shoots[J].Plant Physiology Journal,52(3):372-380.]
徐秋云,王华磊,赵致,刘洪昌,李金玲,罗春丽,罗夫来,黄明进,熊显榕.2012.何首乌组培快繁培养基的优化[J].贵州农业科学,40(10):7-10.[Xu Q Y,Wang H L,Zhao Z,Liu H C,Li J L,Luo C L,Luo F L,Huang M J,Xiong X R.2012.Optimization on medium for tissue culture and rapid propagation of Polygonum multiforum[J].Guizhou Agricultural Scicences,40(10):7-10.]
许亚良,张家明.2013.一种高效大规模组培方法--间歇浸没培养法[J].植物生理学报,49(4):392-399.[Xu Y L,Zhang J M.2013.An efficient method for mass tissue culture-Temporary immersion culture principle[J].Plant Physiology Journal,49(4):392-399.]
杨柳,秦钢,杨丽涛,罗瑞鸿,游建华,Ariel D Arencibia,魏源文,李杨瑞.2010.利用间歇浸没式生物反应器进行果蔗组培快繁[J].热带作物学报,31(4):614-620.[Yang L,Qin G,Yang L T,Luo R H,You J H,Arencibia A D,Wei Y W,Li Y R.2010.Optimization of saccharum sinensis roxb rapid multiplication in temporary immersion bioreactors system[J].Chinese Journal of Tropical Crops,31(4):614-620.]
姚焱,汪珍春,王小兰,张平,田长恩.2010.广东道地中药何首乌的组织培养[J].北方园艺,(7):175-177.[Yao Y,Wang Z C,Wang X L,Zhang P,Tian C E.2010.Tissue culture of Polygonum multiflorum Thunb origined from Guangdong Deqing[J].Northern Horticulture,(7):175-177.]
袁红霞,王金虎,陈德燕.2005.何首乌块根愈伤组织培养[J].苏州大学学报,21(3):86-88.[Yuan H X,Wang JH,Chen D Y.2005.Callus culture of Polygonum multiforum root tuber[J].Journal of Suzhou University,21(3):86-88.]
曾文丹,陆柳英,谢向誉,严华兵.2014.优良木薯品种新选048离体快繁技术[J].南方农业学报,45(6):950-954.[Zeng W D,Lu L Y,Xie X Y,Yan H B.2014.Rapid propagation in vitro technology for fine cassava variety Xinxuan 048[J].Journal of Southern Agriculture,45(6):950-954.]
曾文丹,陆柳英,谢向誉,严华兵.2016.何首乌繁殖技术研究进展[J].中国热带农业,(3):66-68.[Zeng W D,Lu LY,Xie X Y,Yan H B.2016.Research progress of reproduction of Polygonum multiforum[J].China Tropical Agriculture,(3):66-68.]
朱桥,丁俊伟,杨学文,梁廉,邵玲.2014.血叶兰的组织培养与快速繁殖[J].植物生理学报,50(6):805-809.[Zhu Q,Ding J W,Yang X W,Liang L,Shao L.2014.Tissue culture and rapid propagation of Ludisia discolor[J].Plant Physiology Journal,50(6):805-809.]
Rachel W L,David G L,Stephen P M,Leach D N.2003.Antiinflammatory activity of Chinese medicinal vine plants[J].Journal of Ethnopharmacol,85(1):61.
Yan H B,Liang C X,Li Y R.2010.Improved growth and quality of Siraitia grosvenorii plantlets using a temporary immersion system[J].Plant Cell,Tissue and Organ Culture,103(1):131-135.
Yang P Y,Almofti M R,Lu L,Kang H,Zhang J,Li T J,Rui Y C,Sun L N,Chen W S.2005.Reduction of atherosclerosis in cholesterol fed rabbits and decrease of expression of intracellular adhesion molecule and vascular endothelial growth factor in foam celles by a water-soluble fraction of Polygonum multiflorum Thunb[J].Journal of Pharmacal Science,99(3):294.
陈菊,陈国惠.2006b.6-BA与NAA浓度配比对何首乌不定芽增殖的影响[J].中国农学通报,22(11):173-175.[Chen J,Chen G H.2006b.Study of the concentration of 6-BAand NAA in the rapid propagation of Polygonum multiflorum Thunb[J].Chinese Agricultural Science Bulletin,22(11):173-175.]
高美萍,林志城,张驰,江文,董伟清,欧昆鹏,桂杰,闭志强,何芳练,陈丽娟.2016.荸荠组培苗在间歇浸没式生物反应器(TIBs)中的高效增殖技术[J].南方农业学报,47(10):1653-1657.[Gao M P,Lin Z C,Zhang C,Jiang W,Dong W Q,Ou K P,Gui J,Bi Z Q,He F L,Chen LJ.2016.Efficient propagation of Eleocharis dulcis plantlets in temporary immersion bioreactors(TIBs)[J].Journal of Southern Agriculture,47(10):1653-1657.]
贾梦雪,徐瑾,叶香娟,刘芊,王喆,刘燕.2013.春石斛优良品种‘森禾2006’组培快繁体系的建立[J].植物生理学报,49(12):1363-1367.[Jia M X,Xu J,Ye X J,Liu Q,Wang Z,Liu Y.2013.Establishment of rapid propagation system for elite variety of Dendrobium nobile Lindl.‘Senhe 2006’by tissue culture[J].Plant Physiology Journal,49(12):1363-1367.]
李晶,罗玉洁,张青林,罗正荣,刘继红.2016.君迁子休眠芽及叶片离体培养体系优化及植株再生[J].华中农业大学学报,35(4):14-19.[Li J,Luo Y J,Zhang Q L,Luo Z R,Liu J H.2016.In vitro culture system optimization and regeneration of date plum(Diospyros lotus Linn.)dormant buds and leaves[J].Journal of Huazhong Agricultural University,35(4):14-19.]
李娟玲,刘国民,邱文华,徐立新.2003.何首乌茎段离体培养的研究[J].贵州科学,21(3):86-91.[Li J L,Liu G M,Qiu W H,Xu L X.2003.Study on the stem segments culture in vitro of Polygonum multiflorum Thunb[J].Guizhou Science,21(3):86-91.]
刘义存,黄天启,林顺权.2017.枇杷属植物野生种的茎段愈伤组织诱导植株再生研究[J].果树学报,34(6):762-768.[Liu Y C,Huang T Q,Lin S Q.2017.Study on plant regeneration of callus from the stem of wild loquat[J].Journal of Fruit Science,34(6):762-768.]
龙滢,杨鑫,余春香.2005.何首乌的组织培养研究[J].中医药导报,11(11):63-64.[Long Y,Yang X,Yu C X.2005.Tissue culture research on fleece flower root[J].Guiding Journal of TCM,11(11):63-64.]
孙桂波,纪凤兰,徐惠波,孙晓波.2007.何首乌的化学成分与药理作用研究进展[J].长春中医药大学学报,23(4):105.[Sun G B,Ji F L,Xu H B,Sun X B.2007.Advances in studies on the chemical components and pharmacological activities of Radix polygoni multiflora[J].Journal of Changchun University of Traditional Chinese Medicine,23(4):105.]
孙英坤,胡绍庆,庞基良,高凯,刘华红,陈焕伟,姚涛,陈林敬,沈柏春.2017.珍稀濒危物种堇叶紫金牛高效快繁体系的建立[J].植物学报,52(6):764-773.[Sun Y K,Hu S Q,Pang J L,Gao K,Liu H H,Chen H W,Yao T,Chen L J,Shen B C.2017.Establishment of a tissue culture and propagation system for Ardisia violacea,a rare and endangered species[J].Chinese Bulletin of Botany,52(6):764-773.]
魏岳荣,杨护,黄秉智,黄霞,黄学林,邱继水,许林兵.2007.Picloram、ABA和TDZ对香蕉体细胞胚胎发生的影响[J].园艺学报,34(1):81-86.[Wei Y R,Yang H,Huang B Z,Huang X,Huang X L,Qiu J S,Xu L B.2007.Effects of Picloram,ABA and TDZ on somatic embryogenesis of banana[J].Acta Horticulturae Sinica,34(1):81-86.]
吴光洪,孙英坤,陈林敬,沈柏春,邓梦楚,钱飞.2016.兔眼蓝莓‘粉蓝’叶片直接诱导丛生芽再生技术体系的建立[J].植物生理学报,52(3):372-380.[Wu G H,Sun Y C,Chen L J,Shen B C,Deng M C,Qian F.2016.The technology system establishment for direct induction of Vaccinium ashei‘Powderblue’leaves to regenerate multiple shoots[J].Plant Physiology Journal,52(3):372-380.]
徐秋云,王华磊,赵致,刘洪昌,李金玲,罗春丽,罗夫来,黄明进,熊显榕.2012.何首乌组培快繁培养基的优化[J].贵州农业科学,40(10):7-10.[Xu Q Y,Wang H L,Zhao Z,Liu H C,Li J L,Luo C L,Luo F L,Huang M J,Xiong X R.2012.Optimization on medium for tissue culture and rapid propagation of Polygonum multiforum[J].Guizhou Agricultural Scicences,40(10):7-10.]
许亚良,张家明.2013.一种高效大规模组培方法--间歇浸没培养法[J].植物生理学报,49(4):392-399.[Xu Y L,Zhang J M.2013.An efficient method for mass tissue culture-Temporary immersion culture principle[J].Plant Physiology Journal,49(4):392-399.]
杨柳,秦钢,杨丽涛,罗瑞鸿,游建华,Ariel D Arencibia,魏源文,李杨瑞.2010.利用间歇浸没式生物反应器进行果蔗组培快繁[J].热带作物学报,31(4):614-620.[Yang L,Qin G,Yang L T,Luo R H,You J H,Arencibia A D,Wei Y W,Li Y R.2010.Optimization of saccharum sinensis roxb rapid multiplication in temporary immersion bioreactors system[J].Chinese Journal of Tropical Crops,31(4):614-620.]
姚焱,汪珍春,王小兰,张平,田长恩.2010.广东道地中药何首乌的组织培养[J].北方园艺,(7):175-177.[Yao Y,Wang Z C,Wang X L,Zhang P,Tian C E.2010.Tissue culture of Polygonum multiflorum Thunb origined from Guangdong Deqing[J].Northern Horticulture,(7):175-177.]
袁红霞,王金虎,陈德燕.2005.何首乌块根愈伤组织培养[J].苏州大学学报,21(3):86-88.[Yuan H X,Wang JH,Chen D Y.2005.Callus culture of Polygonum multiforum root tuber[J].Journal of Suzhou University,21(3):86-88.]
曾文丹,陆柳英,谢向誉,严华兵.2014.优良木薯品种新选048离体快繁技术[J].南方农业学报,45(6):950-954.[Zeng W D,Lu L Y,Xie X Y,Yan H B.2014.Rapid propagation in vitro technology for fine cassava variety Xinxuan 048[J].Journal of Southern Agriculture,45(6):950-954.]
曾文丹,陆柳英,谢向誉,严华兵.2016.何首乌繁殖技术研究进展[J].中国热带农业,(3):66-68.[Zeng W D,Lu LY,Xie X Y,Yan H B.2016.Research progress of reproduction of Polygonum multiforum[J].China Tropical Agriculture,(3):66-68.]
朱桥,丁俊伟,杨学文,梁廉,邵玲.2014.血叶兰的组织培养与快速繁殖[J].植物生理学报,50(6):805-809.[Zhu Q,Ding J W,Yang X W,Liang L,Shao L.2014.Tissue culture and rapid propagation of Ludisia discolor[J].Plant Physiology Journal,50(6):805-809.]
Rachel W L,David G L,Stephen P M,Leach D N.2003.Antiinflammatory activity of Chinese medicinal vine plants[J].Journal of Ethnopharmacol,85(1):61.
Yan H B,Liang C X,Li Y R.2010.Improved growth and quality of Siraitia grosvenorii plantlets using a temporary immersion system[J].Plant Cell,Tissue and Organ Culture,103(1):131-135.
Yang P Y,Almofti M R,Lu L,Kang H,Zhang J,Li T J,Rui Y C,Sun L N,Chen W S.2005.Reduction of atherosclerosis in cholesterol fed rabbits and decrease of expression of intracellular adhesion molecule and vascular endothelial growth factor in foam celles by a water-soluble fraction of Polygonum multiflorum Thunb[J].Journal of Pharmacal Science,99(3):294.