阳春砂愈伤组织的诱导
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摘要
目的:通过阳春砂组织培养获得愈伤组织,建立阳春砂愈伤组织的诱导体系。方法:将阳春砂根状茎芽及阳春砂试管苗的茎段、根尖作为愈伤组织诱导的外植体材料,接种到以MS为基本培养基,分别添加不同浓度的6-苄基腺嘌呤(6-BA),ɑ-萘乙酸(NAA)及2,4-二氯苯氧乙酸(2,4-D)的培养基中(各培养基的p H约为5. 8),探讨不同外植体材料及不同培养基对阳春砂愈伤组织诱导的影响。结果:研究结果显示,阳春砂根状茎芽及试管苗的茎段、根尖3种外植体材料均能有效地诱导产生愈伤组织;其中阳春砂根状茎芽、试管苗茎段2种外植体材料诱导产生愈伤组织的最适培养基为MS+6-BA(1. 5 mg·L-1)+2,4-D(1. 0 mg·L-1)+NAA(0. 5 mg·L-1),最高诱导率分别为15%和60%;试管苗根尖诱导产生愈伤组织的最适培养基为MS+6-BA(2. 0 mg·L-1)+2,4-D(1. 0 mg·L-1)+NAA(1. 0 mg·L-1),最高诱导率76%,为阳春砂愈伤组织诱导的最佳外植体材料。结论:该研究初步建立了阳春砂的根状茎芽及试管苗的茎段、根尖3种外植体的愈伤组织诱导体系,试管苗根尖为愈伤组织诱导的最佳外植体。
Objective: To set up a callus induction system for Amomum villosum by tissue culture.Method: The rhizome buds of A. villosum and stem segments,root tip segments of sterile A. villosum plantles were used as explants and cultured in MS media with different concentrations of 6-BA,NAA and 2,4-D( the p H of each medi is about 5. 8). A callus induction system was established to explore the effect of different explants and different medium on callus induction for A. villosum. Result: The findings showed that the rhizome buds and sterile plantlet stems and root tip segments of three different explants can be successfully induced into calli. The most suitable medium for callus induction from rhizome buds and sterile plantlet stems was MS with 6-BA( 1. 5 mg·L-1),2,4-D( 1. 0 mg·L-1) and NAA( 0. 5 mg·L-1) with the highest induction rates of 15% and 60%respectively. MS medium combined with 6-BA( 2. 0 mg·L-1),2,4-D( 1. 0 mg·L-1) and NAA( 1. 0 mg·L-1)was the most suitable proposal for inducing the callus from sterile root tip segments with the highest induction rate of76%. Conclusion: Under certain culture conditions,rhizome buds,stem or root tip segments of sterile plantlet can be effectively induced into callus. The callus induction system of A. villosum is preliminarily established,and root tip segments of sterile plantlet are the optimal explant.
Objective: To set up a callus induction system for Amomum villosum by tissue culture.Method: The rhizome buds of A. villosum and stem segments,root tip segments of sterile A. villosum plantles were used as explants and cultured in MS media with different concentrations of 6-BA,NAA and 2,4-D( the p H of each medi is about 5. 8). A callus induction system was established to explore the effect of different explants and different medium on callus induction for A. villosum. Result: The findings showed that the rhizome buds and sterile plantlet stems and root tip segments of three different explants can be successfully induced into calli. The most suitable medium for callus induction from rhizome buds and sterile plantlet stems was MS with 6-BA( 1. 5 mg·L-1),2,4-D( 1. 0 mg·L-1) and NAA( 0. 5 mg·L-1) with the highest induction rates of 15% and 60%respectively. MS medium combined with 6-BA( 2. 0 mg·L-1),2,4-D( 1. 0 mg·L-1) and NAA( 1. 0 mg·L-1)was the most suitable proposal for inducing the callus from sterile root tip segments with the highest induction rate of76%. Conclusion: Under certain culture conditions,rhizome buds,stem or root tip segments of sterile plantlet can be effectively induced into callus. The callus induction system of A. villosum is preliminarily established,and root tip segments of sterile plantlet are the optimal explant.
引文
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[7]徐杰,李明晓,苏景,等.阳春砂-龙眼生态立体种植模式的研究[J].中国中药杂志,2018,43(2):288-298.
[8]蒋烨,苏景,汤丽云,等.一种阳春砂新栽培类型的鉴定[J].广西植物,2017,37(5):554-564.
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[11]刁玲武,董燕,周联,等.阳春砂愈伤组织诱导的多因子正交试验研究[J].广州中医药大学学报,2011,28(4):434-438,460.
[12]付航,黄涵签,王妍,等.防风愈伤组织诱导及植株再生体系的建立[J].中草药,2018,49(13):3127-3133.
[13]徐瑞超,马云桐,陈新,等.三角叶黄连愈伤组织诱导和培养条件的优化[J].中国实验方剂学杂志,2012,18(1):113-116.
[14] Wong K F,Taha R M. The effect of 2,4-dichlorophenoxyacetic acid and 6-benzylaminopurine on callus induction and plant regeneration of Allamanda cathartica-A valuable medicinal plant[J]. Res J Biotechnol,2012,7(3):75-80.
[15]游建军,彭建明,李荣英,等.植物生长调节物质对白豆蔻花粉萌发和花粉管生长的影响[J].中国实验方剂学杂志,2012,18(22):172-175.
[16]周宜君,周生闯,刘玉,等.植物生长调节剂对植物愈伤组织的诱导与分化的影响[J].中央民族大学学报,2007,16(1):23-28.
[17]李俊强,林丽华,张帆,等.早开堇菜组织培养及植株再生体系的建立[J].草业学报,2015,24(11):163-173.
[18]张林甦,赵德刚.玄参愈伤、不定根和内生菌产哈巴俄苷的比较[J].中国实验方剂学杂志,2012,18(20):136-139.
[2]国家药典委员会.中华人民共和国药典.一部[M].北京:中国医药科技出版社,2015:253.
[3]段立胜,张丽霞,彭建明,等.西双版纳阳春砂仁种质资源调查初报[J].时珍国医国药,2009,20(3):627-628.
[4]高伟.药用植物阳春砂生殖生物学特性初步研究[D].广州:广州中医药大学,2011.
[5]陆善旦,王健.春砂仁销势连年看好[J].中国现代中药,2008,10(2):48-49.
[6]汤丽云,何国振,苏景,等.道地春砂仁产业发展的策略研究[J].中国农学通报,2012,28(8):94-99.
[7]徐杰,李明晓,苏景,等.阳春砂-龙眼生态立体种植模式的研究[J].中国中药杂志,2018,43(2):288-298.
[8]蒋烨,苏景,汤丽云,等.一种阳春砂新栽培类型的鉴定[J].广西植物,2017,37(5):554-564.
[9]肖尊安.植物生物技术[M].北京:化学工业出版社,2005:31.
[10]张雅明,董燕,周联,等.阳春砂愈伤组织诱导与植株再生[J].广州中医药大学学报,2007,24(1):66-68.
[11]刁玲武,董燕,周联,等.阳春砂愈伤组织诱导的多因子正交试验研究[J].广州中医药大学学报,2011,28(4):434-438,460.
[12]付航,黄涵签,王妍,等.防风愈伤组织诱导及植株再生体系的建立[J].中草药,2018,49(13):3127-3133.
[13]徐瑞超,马云桐,陈新,等.三角叶黄连愈伤组织诱导和培养条件的优化[J].中国实验方剂学杂志,2012,18(1):113-116.
[14] Wong K F,Taha R M. The effect of 2,4-dichlorophenoxyacetic acid and 6-benzylaminopurine on callus induction and plant regeneration of Allamanda cathartica-A valuable medicinal plant[J]. Res J Biotechnol,2012,7(3):75-80.
[15]游建军,彭建明,李荣英,等.植物生长调节物质对白豆蔻花粉萌发和花粉管生长的影响[J].中国实验方剂学杂志,2012,18(22):172-175.
[16]周宜君,周生闯,刘玉,等.植物生长调节剂对植物愈伤组织的诱导与分化的影响[J].中央民族大学学报,2007,16(1):23-28.
[17]李俊强,林丽华,张帆,等.早开堇菜组织培养及植株再生体系的建立[J].草业学报,2015,24(11):163-173.
[18]张林甦,赵德刚.玄参愈伤、不定根和内生菌产哈巴俄苷的比较[J].中国实验方剂学杂志,2012,18(20):136-139.
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