玄参组织培养及炼苗技术
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摘要
目的:以玄参茎段、叶柄、带腋茎段和叶片为外植体,筛选出适宜的植株再生体系以及最佳的炼苗条件。方法:通过L_(16)(4~5)正交试验研究玄参不同外植体、不同激素和不同浓度对丛生芽的诱导和增殖;采用单因素方差分析不同浓度激素对不定芽生根的诱导;同时选择不同基质、不同浇水周期和不同过渡方式对玄参生根苗炼苗关键技术的优选。结果:带腋茎段为最佳外植体,其次依次为茎段,叶片和叶柄;其在MS+6-苄氨基腺嘌呤(6-BA) 0. 5 mg·L~(-1)+萘乙酸(NAA) 0. 2 mg·L~(-1)中不定芽诱导率和芽增殖倍数分别为100%和9. 84;适宜不定芽生根的培养基为1/2 MS+吲哚-3-丁酸(IBA) 0. 2 mg·L~(-1),生根率100%,根数为39. 45条;就基质而言,以营养土-蛭石-珍珠岩(5∶2∶1)为移栽基质,可保持基质中肥力、透气性与保水性适当搭配;浇水周期选择每2 d浇1次水,过渡方式为室内炼苗20 d,室外遮阴大棚炼苗10 d后移栽大田,成活率可达到95%以上。结论:建立玄参无性快速繁殖体系,为提供玄参高效、快速、稳定的植株再生,优质的种苗繁育以及适宜的炼苗条件提供了有效途径。
Objective: Stems, petioles, stem sections with axillary and leaves of Scrophularia ningpoensis were taken as the material in vitro to screen out the suitable plantlet regeneration system and optimal exercising seedling conditions. Method: Different explants,hormones and concentrations on the induction and proliferation of cluster bud were studied by L_(16)(4~5) orthogonal test. One factor analysis of variance(ANOVA)was made on the induction of adventitious buds rooted with different concentrations of hormones. At the same time,different substrates,watering cycles and transition modes were selected to optimize key technologies of exercising seedlings of S. ningpoensis. Result: Stem sections with axillary was the best explant,which was followed by stems,leaves and petioles. The suitable media for the induction of adventitious buds was MS + 6-BA0. 5 mg·L~(-1)+ NAA 0. 2 mg·L~(-1),with the induction rate of 100. 0% and the proliferation multiple of 9. 84. The suitable media for root induction was 1/2 MS + IBA 0. 2 mg·L~(-1),with the rooting rate of 100. 0% and the number of roots of 39. 45. For matrix,they were transplanted with nutrient soil,vermiculite and perlite(5 ∶ 2 ∶ 1) as the media,to keep proper matching of fertility,permeability and water retention. The container seedlings can grow well,and the survival rate was more than 95% when they were watered every 2 days, the acclimatization of plantlets took 20 days indoor and 10 days in shaded greenhouses. Conclusion: The clonal propagation system of S. ningpoensis was established to provide an effective way for the efficient,rapid and steady plantlet regeneration,the breeding of high-quality seedlings and the suitable exercising seedling conditions of S. ningpoensis.
Objective: Stems, petioles, stem sections with axillary and leaves of Scrophularia ningpoensis were taken as the material in vitro to screen out the suitable plantlet regeneration system and optimal exercising seedling conditions. Method: Different explants,hormones and concentrations on the induction and proliferation of cluster bud were studied by L_(16)(4~5) orthogonal test. One factor analysis of variance(ANOVA)was made on the induction of adventitious buds rooted with different concentrations of hormones. At the same time,different substrates,watering cycles and transition modes were selected to optimize key technologies of exercising seedlings of S. ningpoensis. Result: Stem sections with axillary was the best explant,which was followed by stems,leaves and petioles. The suitable media for the induction of adventitious buds was MS + 6-BA0. 5 mg·L~(-1)+ NAA 0. 2 mg·L~(-1),with the induction rate of 100. 0% and the proliferation multiple of 9. 84. The suitable media for root induction was 1/2 MS + IBA 0. 2 mg·L~(-1),with the rooting rate of 100. 0% and the number of roots of 39. 45. For matrix,they were transplanted with nutrient soil,vermiculite and perlite(5 ∶ 2 ∶ 1) as the media,to keep proper matching of fertility,permeability and water retention. The container seedlings can grow well,and the survival rate was more than 95% when they were watered every 2 days, the acclimatization of plantlets took 20 days indoor and 10 days in shaded greenhouses. Conclusion: The clonal propagation system of S. ningpoensis was established to provide an effective way for the efficient,rapid and steady plantlet regeneration,the breeding of high-quality seedlings and the suitable exercising seedling conditions of S. ningpoensis.
引文
[1]刘树民,张宁,于卉,等.玄参性味的本草考证[J].中华中医药杂志,2017,32(2):429-431.
[2]国家药典委员会.中华人民共和国中国药典一部.[M].北京:中国医药科技出版社,2015:117.
[3]许福泉,许旭东,陈士林.玄参化学成分及药理活性研究进展[J].中国现代中药,2013,15(9):752-759.
[4]俞静静,陈素红,吕圭源.玄参“凉血滋阴”药效相关研究概况[J].中国实验方剂学杂志,2007,13(9):63-66.
[5]YANG S T,CHEN C,ZHAO Y P,et al.Association between chemical and genetic variation of wild and cultivated populations of Scrophularia ningpoensis Hemsl[J].Planta Med,2011,77(8):865-871.
[6]王泽均.玄参组织培养植株再生[J].植物生理学通讯,1983(6):42.
[7]李浚明.植物组织培养教程[M].北京:中国农业大学出版社,1996:345.
[8]陈敏艳,梁宗锁,王喆之,等.地黄组织培养及植株再生的研究[J].西北植物学报,2004,24(6):1083-1087.
[9]任如意,薛巨坤,李艳萍,等.药用植物北玄参高频再生体系建立[J].植物生理学报,2014,50(12):1816-1820.
[10]任继文,雷颖,李晓玲.款冬叶柄愈伤组织培养与再生体系建立[J].中国中药杂志,2017,42(20):3895-3900.
[11]WANG C L,DONG X N,DING M Q,et al.Plantlet regeneration of tartary buckwheat(Fagopyrum tataricum Gaertn.)in vitro tissue cultures[J].Protein Pept Lett,2016,23(5):468-477.
[12]任如意,薛巨坤,国会艳,等.北玄参毛状根诱导及其植株再生[J].植物学报,2017,52(6):783-787.
[13]吴秀华,张艳玲,周月,等.‘海沃德’猕猴桃叶片高频直接再生体系的建立[J].植物生理学报,2013,49(8):759-763.
[14]朱桥,丁俊伟,杨学文,等.血叶兰的组织培养与快速繁殖[J].植物生理学报,2014,50(6):805-809.
[15]Thaniarasu R,Senthil Kumar T,RAO M V,et al.Mass propagation of Plectranthus bourneae Gamble through indirect organogenesis from leaf and internode explants[J].Physiol Mol Biol Plants,2016,22(1):143-151.
[2]国家药典委员会.中华人民共和国中国药典一部.[M].北京:中国医药科技出版社,2015:117.
[3]许福泉,许旭东,陈士林.玄参化学成分及药理活性研究进展[J].中国现代中药,2013,15(9):752-759.
[4]俞静静,陈素红,吕圭源.玄参“凉血滋阴”药效相关研究概况[J].中国实验方剂学杂志,2007,13(9):63-66.
[5]YANG S T,CHEN C,ZHAO Y P,et al.Association between chemical and genetic variation of wild and cultivated populations of Scrophularia ningpoensis Hemsl[J].Planta Med,2011,77(8):865-871.
[6]王泽均.玄参组织培养植株再生[J].植物生理学通讯,1983(6):42.
[7]李浚明.植物组织培养教程[M].北京:中国农业大学出版社,1996:345.
[8]陈敏艳,梁宗锁,王喆之,等.地黄组织培养及植株再生的研究[J].西北植物学报,2004,24(6):1083-1087.
[9]任如意,薛巨坤,李艳萍,等.药用植物北玄参高频再生体系建立[J].植物生理学报,2014,50(12):1816-1820.
[10]任继文,雷颖,李晓玲.款冬叶柄愈伤组织培养与再生体系建立[J].中国中药杂志,2017,42(20):3895-3900.
[11]WANG C L,DONG X N,DING M Q,et al.Plantlet regeneration of tartary buckwheat(Fagopyrum tataricum Gaertn.)in vitro tissue cultures[J].Protein Pept Lett,2016,23(5):468-477.
[12]任如意,薛巨坤,国会艳,等.北玄参毛状根诱导及其植株再生[J].植物学报,2017,52(6):783-787.
[13]吴秀华,张艳玲,周月,等.‘海沃德’猕猴桃叶片高频直接再生体系的建立[J].植物生理学报,2013,49(8):759-763.
[14]朱桥,丁俊伟,杨学文,等.血叶兰的组织培养与快速繁殖[J].植物生理学报,2014,50(6):805-809.
[15]Thaniarasu R,Senthil Kumar T,RAO M V,et al.Mass propagation of Plectranthus bourneae Gamble through indirect organogenesis from leaf and internode explants[J].Physiol Mol Biol Plants,2016,22(1):143-151.