‘宁糯芋1号’茎尖离体培养条件优化
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摘要
为丰富福建省优质芋种源,加快优良品种的推广应用,以‘宁糯芋1号’块茎顶芽和侧芽为外植体,运用茎尖分生组织培养获得了不定芽。在正交试验基础上,根据响应面分析法中心组合试验,以不定芽增殖系数为响应值,对TDZ、温度2个因素进行优化。结果发现,与6-BA相比,TDZ更适于芋茎尖诱导丛芽,在MS培养基中附加0.1～0.2 mg/L的TDZ能显著促进芋茎尖的萌动,成苗率达76.7%～80.0%。经芋花叶病毒(DMV)检测试剂盒的ELISA检测表明,诱导芽的脱毒率为90%;响应面优化结果表明,‘宁糯芋1号’不定芽继代增殖的最佳培养条件为MS+TDZ 0.31 mg/L,培养温度30.32℃,增殖系数预测值为9.08,验证得到的实际值为8.86。利用响应面方法可有效优化‘宁糯芋1号’不定芽继代增殖培养条件。
To enrich the taro germplasms in Fujian Province and accelerate the application of elite varieties,using the apical bud and lateral bud of‘1~(st) Ning waxy taro'tuber as explants, adventitious buds were induced from apical meristem culture. Based on the orthogonal experiment, the two factors of TDZ concentration and temperature were optimized using adventitious bud multiplication coefficient as a response value with central composite design of response surface method. The results showed that TDZ was more appropriate for the clustered buds induction from apical meristem, compared with 6-BA. The bud germination was significantly promoted on MS medium when supplemented with 0.1-0.2 mg/L TDZ, and the planting rates were 76.7%-80.0%. ELISA of taro mosaic virus(DMV) detection kits showed that the detoxification rate of induced bud was90%. The response surface method showed that the optimal culture condition of adventitious buds subculture multiplication was MS +TDZ 0.31 mg/L, and the culture temperature was 30.32℃. The predicted value of multiplication coefficient was 9.08, while the actual value was 8.86. Therefore, the response surface method can effectively optimize the subculture multiplication condition of adventitious buds from‘1~(st) Ning waxy taro'.
To enrich the taro germplasms in Fujian Province and accelerate the application of elite varieties,using the apical bud and lateral bud of‘1~(st) Ning waxy taro'tuber as explants, adventitious buds were induced from apical meristem culture. Based on the orthogonal experiment, the two factors of TDZ concentration and temperature were optimized using adventitious bud multiplication coefficient as a response value with central composite design of response surface method. The results showed that TDZ was more appropriate for the clustered buds induction from apical meristem, compared with 6-BA. The bud germination was significantly promoted on MS medium when supplemented with 0.1-0.2 mg/L TDZ, and the planting rates were 76.7%-80.0%. ELISA of taro mosaic virus(DMV) detection kits showed that the detoxification rate of induced bud was90%. The response surface method showed that the optimal culture condition of adventitious buds subculture multiplication was MS +TDZ 0.31 mg/L, and the culture temperature was 30.32℃. The predicted value of multiplication coefficient was 9.08, while the actual value was 8.86. Therefore, the response surface method can effectively optimize the subculture multiplication condition of adventitious buds from‘1~(st) Ning waxy taro'.
引文
[1]黄新芳,柯卫东,叶元英,等.中国芋种质资源研究进展[J].植物遗传资源学报,2005,6(1):119-123.
[2]李宗战.福鼎槟榔芋煮不烂原因与高优栽培措施[J].福建农业科技,2010(5):28-29.
[3]曹静,邝哲师,周丽侬.芋的组培快繁技术[J].植物杂志,1997(1):26.
[4]崔瑾,李式军;吴琴生.芋脱毒快繁体系的构建以及组培苗无糖培养的研究[D].南京:南京农业大学,2002.
[5]刘玉平,柯卫东,黄新芳,等.试管芋诱导的研究[J].园艺学报,2003(1):43-46.
[6]杨俊慧,孟庆军,李建东.等.芋的脱毒快繁及栽培研究[J].山东科学,2004,3:32-35.
[7]龙春林,程治英,王俐,等.李氏香芋的组织培养[J].植物生理学通讯,2004,40(3):339-339.
[8]姜秀芳,张改英,田炜,等.芋茎尖分生组织培养的研究[J].园艺学报,2004(4):466.
[9]汤青林,王小佳,宋明.等.组培芋苗不同世代的差异研究[J].西南大学学报:自然科学版,2007(2):66-69.
[10]赵建萍,蒋小满,柏新富,等.影响芋茎尖培养中脱毒和快速繁殖的几个生理因素[J].植物生理学通讯,2009,45(2):133-136.
[11]王陈,陆忠恒,王建国.等.芋脱毒组培苗培养技术研究[J].现代农业科技,2011(12):109-110.
[12]周明强,周正邦,欧珍贵.等.芭蕉芋良种组培及块茎快繁技术研究[J].贵州农业科学,2011(10):47-50.
[13]王莹,姜永平,杨凯.香沙芋新品种“泰芋一号”脱毒组培技术研究[J].现代园艺,2015(24):7-8.
[14]韩晓勇,宋婷婷,王立,等.靖江香沙芋组织培养快繁技术[J].江苏农业科学,2015,43(1):50-52.
[15]郭克婷,何金明.张溪香芋茎尖组培快繁技术研究[J].广东农业科学,2015,42(6):25-29.
[16]奥斯伯,布伦特,金斯顿,等,著,颜子颖,王海林译.精编分子生物学实验指南[M].北京:科学出版社,1998:414-417.
[17]施世明,洪霓.芋病毒病研究进展[J].长江蔬菜,2010(14):3-5.
[18]陈集双,李德葆.我国13种天南星科作物上的芋花叶病毒[J].微生物学报,1996,36(2):126-131,160.
[19]刘文洪,陈集双,李永伟.侵染天南星科植物病毒的分子鉴定及其生态学研究[J].应用生态学报,2004,15(4):566-570.
[20]Zettler F W,Foxe M J,Hartman R D,et al.Filamentous viruses infecting dasheen and other araceous plants[J].Phytopathology,1970,60:983-987.
[21]韦传宝,贾玉成,杜宇.芋花叶病毒检测试剂盒的研制[J].安徽大学学报:自然科学版,2009,33(4):85-89,94.
[22]周俊彦,郭扶兴.苯基脲衍生物的细胞分裂素活性[J].植物生理学通讯,1990,26(4):7-12.
[23]Mok M C,Mok D W S,Armsstrong D J,et al.Cytokin activity of N-phenyl-N'-1,2,3-thidiazol-5-ylurea(thidiazuron)[J].phytochemistry,1982,21:1509-1511.
[24]Thomas J C,Katterman F R.Cytokinin activity induced by thidiaxuron[J].Plant Physiol,1986,81:681-683.
[25]Capella S C,Mok D W,Kirchner S C,et al.Effect of thidiaxuron on cytokinin autonomy and metabolism of N6-(?2-Isopentenyl)8-14C adenosine in callus tissues of Phaseohus lunatus L.[J].Plant Physiol,1983,73:796-802.
[26]张志宏,景士西,王关林.TDZ对苹果叶片离体再生不定芽的效应[J].植物生理学通讯,1997,33(6):420-423.
[27]崔瑾,朱月林,李式军.苯基噻二唑(TDZ)在不同基因型芋组织培养中的生理效应[J].南京农业大学学报,2002,25(2):31-34.
[28]Chad H,Pearson M N,Lovell P H.Rapid vegetative multiplication in Colocasia esculenta(L).schott(taro)[J].Plant Cell,Tissue,and Organ Culture,1998-1999,55(3):223-226.
[29]田文忠,Iaan Rance,Elunialai Sivamani,等.提高籼稻愈伤组织再生频率的研究[J].遗传学报,1994,21(3):215-222,254.
[30]Chalupa V.Large scale micropropogation of Quercus robur L.using adenine-type cytokinins and thidiazuron to stimulate shoot proliferation[J].Biol Plant(Praha),1988,30:414-421.
[31]潘雷,谷良贤,阎代维.改进响应面法及其近似性能研究[J].宇航学报,2009,30(2):806-810.
[2]李宗战.福鼎槟榔芋煮不烂原因与高优栽培措施[J].福建农业科技,2010(5):28-29.
[3]曹静,邝哲师,周丽侬.芋的组培快繁技术[J].植物杂志,1997(1):26.
[4]崔瑾,李式军;吴琴生.芋脱毒快繁体系的构建以及组培苗无糖培养的研究[D].南京:南京农业大学,2002.
[5]刘玉平,柯卫东,黄新芳,等.试管芋诱导的研究[J].园艺学报,2003(1):43-46.
[6]杨俊慧,孟庆军,李建东.等.芋的脱毒快繁及栽培研究[J].山东科学,2004,3:32-35.
[7]龙春林,程治英,王俐,等.李氏香芋的组织培养[J].植物生理学通讯,2004,40(3):339-339.
[8]姜秀芳,张改英,田炜,等.芋茎尖分生组织培养的研究[J].园艺学报,2004(4):466.
[9]汤青林,王小佳,宋明.等.组培芋苗不同世代的差异研究[J].西南大学学报:自然科学版,2007(2):66-69.
[10]赵建萍,蒋小满,柏新富,等.影响芋茎尖培养中脱毒和快速繁殖的几个生理因素[J].植物生理学通讯,2009,45(2):133-136.
[11]王陈,陆忠恒,王建国.等.芋脱毒组培苗培养技术研究[J].现代农业科技,2011(12):109-110.
[12]周明强,周正邦,欧珍贵.等.芭蕉芋良种组培及块茎快繁技术研究[J].贵州农业科学,2011(10):47-50.
[13]王莹,姜永平,杨凯.香沙芋新品种“泰芋一号”脱毒组培技术研究[J].现代园艺,2015(24):7-8.
[14]韩晓勇,宋婷婷,王立,等.靖江香沙芋组织培养快繁技术[J].江苏农业科学,2015,43(1):50-52.
[15]郭克婷,何金明.张溪香芋茎尖组培快繁技术研究[J].广东农业科学,2015,42(6):25-29.
[16]奥斯伯,布伦特,金斯顿,等,著,颜子颖,王海林译.精编分子生物学实验指南[M].北京:科学出版社,1998:414-417.
[17]施世明,洪霓.芋病毒病研究进展[J].长江蔬菜,2010(14):3-5.
[18]陈集双,李德葆.我国13种天南星科作物上的芋花叶病毒[J].微生物学报,1996,36(2):126-131,160.
[19]刘文洪,陈集双,李永伟.侵染天南星科植物病毒的分子鉴定及其生态学研究[J].应用生态学报,2004,15(4):566-570.
[20]Zettler F W,Foxe M J,Hartman R D,et al.Filamentous viruses infecting dasheen and other araceous plants[J].Phytopathology,1970,60:983-987.
[21]韦传宝,贾玉成,杜宇.芋花叶病毒检测试剂盒的研制[J].安徽大学学报:自然科学版,2009,33(4):85-89,94.
[22]周俊彦,郭扶兴.苯基脲衍生物的细胞分裂素活性[J].植物生理学通讯,1990,26(4):7-12.
[23]Mok M C,Mok D W S,Armsstrong D J,et al.Cytokin activity of N-phenyl-N'-1,2,3-thidiazol-5-ylurea(thidiazuron)[J].phytochemistry,1982,21:1509-1511.
[24]Thomas J C,Katterman F R.Cytokinin activity induced by thidiaxuron[J].Plant Physiol,1986,81:681-683.
[25]Capella S C,Mok D W,Kirchner S C,et al.Effect of thidiaxuron on cytokinin autonomy and metabolism of N6-(?2-Isopentenyl)8-14C adenosine in callus tissues of Phaseohus lunatus L.[J].Plant Physiol,1983,73:796-802.
[26]张志宏,景士西,王关林.TDZ对苹果叶片离体再生不定芽的效应[J].植物生理学通讯,1997,33(6):420-423.
[27]崔瑾,朱月林,李式军.苯基噻二唑(TDZ)在不同基因型芋组织培养中的生理效应[J].南京农业大学学报,2002,25(2):31-34.
[28]Chad H,Pearson M N,Lovell P H.Rapid vegetative multiplication in Colocasia esculenta(L).schott(taro)[J].Plant Cell,Tissue,and Organ Culture,1998-1999,55(3):223-226.
[29]田文忠,Iaan Rance,Elunialai Sivamani,等.提高籼稻愈伤组织再生频率的研究[J].遗传学报,1994,21(3):215-222,254.
[30]Chalupa V.Large scale micropropogation of Quercus robur L.using adenine-type cytokinins and thidiazuron to stimulate shoot proliferation[J].Biol Plant(Praha),1988,30:414-421.
[31]潘雷,谷良贤,阎代维.改进响应面法及其近似性能研究[J].宇航学报,2009,30(2):806-810.