欧洲卫矛叶片愈伤组织诱导及培养
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摘要
以欧洲卫矛(Euonymus europaea)叶片为外植体,研究不同消毒时间对叶片消毒的影响,不同激素种类及质量浓度对愈伤组织诱导、增殖及分化的影响。结果表明:75%酒精30 s+3%次氯酸钠18 min消毒效果最好,污染率最低,为18.74%。6-BA(6-苄氨基嘌呤)对欧洲卫矛叶片愈伤组织诱导的效果好于TDZ(噻苯隆),愈伤组织质量更优。适宜作欧洲卫矛诱导培养基的为MS+1.0 mg·L~(-1)6-BA+0.1 mg·L~(-1)NAA,出愈率达60.7%。适宜作欧洲卫矛增殖培养基的为MS+2.0 mg·L~(-1)6-BA+0.3 mg·L~(-1)NAA+0.1 mg·L~(-1)2,4-D(2,4-二氯苯氧乙酸),增殖率达98.1%,增殖系数达1.953。适宜作欧洲卫矛分化培养基的为MS+1.0 mg·L~(-1)6-BA+0.1 mg·L~(-1)NAA,分化率达47.1%,分化的芽点多。
With the leaves of Euonymus europaea as explants, we studied the effects of different disinfection time on leaf disinfection, and the effects of different hormone types and concentration on callus induction, proliferation and differentiation. The results showed that 75% C_2H_5OH 30 s+3% NaClO for 18 min had the best disinfection effect and the lowest pollution rate was 18.74%. 6-BA on the callus induction of E. europaea leaves better than TDZ. The quality of the callus tissue was better. The suitable medium for callus induction of E. europaea was MS+1.0 mg·L~(-1) 6-BA+0.1 mg·L~(-1) NAA. The recovery rate was 60.7%. The suitable medium for callus proliferation of E. europaea was MS+2.0 mg·L~(-1) 6-BA+0.3 mg·L~(-1) NAA+0.1 mg·L~(-1) 2,4-D. The proliferation rate reached 98.1%, and the proliferation coefficient reached 1.953. The suitable medium foe callus differentiation of E. europaea was MS+1.0 mg·L~(-1) 6-BA+0.1 mg·L~(-1) NAA. The differentiation rate was 47.1% with more differentiated bud points.
With the leaves of Euonymus europaea as explants, we studied the effects of different disinfection time on leaf disinfection, and the effects of different hormone types and concentration on callus induction, proliferation and differentiation. The results showed that 75% C_2H_5OH 30 s+3% NaClO for 18 min had the best disinfection effect and the lowest pollution rate was 18.74%. 6-BA on the callus induction of E. europaea leaves better than TDZ. The quality of the callus tissue was better. The suitable medium for callus induction of E. europaea was MS+1.0 mg·L~(-1) 6-BA+0.1 mg·L~(-1) NAA. The recovery rate was 60.7%. The suitable medium for callus proliferation of E. europaea was MS+2.0 mg·L~(-1) 6-BA+0.3 mg·L~(-1) NAA+0.1 mg·L~(-1) 2,4-D. The proliferation rate reached 98.1%, and the proliferation coefficient reached 1.953. The suitable medium foe callus differentiation of E. europaea was MS+1.0 mg·L~(-1) 6-BA+0.1 mg·L~(-1) NAA. The differentiation rate was 47.1% with more differentiated bud points.
引文
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[20] 孙姿.夏蜡梅组织培养技术研究[D].南京:南京林业大学,2015.
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[22] 聂王星,於丙军.TDZ和6-BA对大豆子叶节再生体系中丛生芽诱导的效应[J].南京农业大学学报,2012,35(4):130-134.
[23] 姜蕾,兰天维,黎扬辉,等.影响红掌愈伤组织诱导、增殖和芽分化的因素[J].种子,2006,25(11):26-30.
[24] 李千,孙丹,李宏博,等.北五味子愈伤组织增殖及胚性愈伤组织诱导的研究[J].种子,2014,33(6):5-9.
[25] 王新,成仿云,钟原,等.凤丹牡丹鳞芽离体培养与快繁技术[J].林业科学,2016,52(5):101-110.
[26] DAS J,MAO A A,HANDIQUE P J.Callus-mediated organogenesis and effect of growth regulators on production of different valepotriates in Indian valerian (Valeriana jatamansi Jones.)[J].Acta Physiologiae Plantarum,2013,35(1):55-63.
[27] 何朋,刘思妤,王宏宇,等.北细辛叶柄愈伤组织的增殖培养及器官分化研究[J].中草药,2016,47(13):2341-2345.
[28] 赵金梅,李芳,周禾,等.紫花苜蓿组织培养体系的建立[J].核农学报,2010,24(3):507-512.
[2] BONNEAU L,BERANGER NOVAT N,MONIN J.Somatic embryogenesis and plant regeneration in a woody species:the european spindle tree (Euonymus europaeus L.)[J].Plant Cell Reports,1994,13(3/4):135-138.
[3] HILL M O,PRESTON C D,ROY D B.Plantatt atributes of british and irish plants:status,size,life history,geography and habitats[J].Centre for Ecology & Hydrology,2004,581(41):5-6.
[4] 赵宇新.欧洲卫矛中新的倍半萜类成分[J].国际中医中药杂志,2003,25(2):100.
[5] 方振峰,华会明.卫矛属植物化学成分及药理活性研究进展[J].现代药物与临床,2007,22(1):6-11.
[6] 蒋金炜,黄翠虹,薛堃,等.秋家蝇对两种卫矛植物的趋性[J].昆虫学报,2008,51(12):1309-1312.
[7] 邢歆,丁彦芬,余义亮.不同品种欧洲卫矛光合特性[J].东北林业大学学报,2018,46(1):12-16.
[8] 王晓花,滕瑶,王德斌,等.提高卫矛种子发芽率的方法[J].林业科技,2003,28(3):4-5.
[9] 赵天鹏,丁彦芬,陈舒博.欧洲卫矛嫁接技术[J].东北林业大学学报,2016,44(7):1-3.
[10] 胡彦,赵艳.植物组织培养技术的应用以及在培养过程中存在的问题[J].陕西师范大学学报(自然科学版),2004(S1):130-134.
[11] 李浚明.植物组织培养教程[M].北京农业大学出版社,2002.
[12] 朱广廉.植物组织培养中的外植体灭菌[J].植物生理学通讯,1996,32(6):444-449.
[13] 胡凯,张立军,白雪梅,等.植物组织培养污染原因分析及外植体的消毒[J].安徽农业科学,2007,35(3):680-681.
[14] 陈正华.木本植物组织培养及其应用[M].北京:高等教育出版社,1986:134-135.
[15] 林妃,李敬阳,常胜合,等.花梨木组织培养外植体消毒方法初步探讨[J].基因组学与应用生物学,2013,32(4):522-525.
[16] 崔美,焦其庆,陈学森,等.苹果叶片愈伤组织的诱导培养[J].山东农业科学,2012,44(3):17-20.
[17] 李琰,姜在民,唐锐.杜仲叶片愈伤组织诱导的激素优化研究[J].植物研究,2006,26(2):182-186.
[18] 徐晓峰,黄学林.TDZ:一种有效的植物生长调节剂[J].植物学报,2003,20(2):227-237.
[19] 白芝兰,李果果,吴晓冉,等.野生毛桃叶片愈伤组织诱导及不定芽再生[J].果树学报,2012,29(5):814-819.
[20] 孙姿.夏蜡梅组织培养技术研究[D].南京:南京林业大学,2015.
[21] 詹亚光,杨传平.白桦愈伤组织的高效诱导和不定芽分化[J].植物生理学报,2002,38(2):111-114.
[22] 聂王星,於丙军.TDZ和6-BA对大豆子叶节再生体系中丛生芽诱导的效应[J].南京农业大学学报,2012,35(4):130-134.
[23] 姜蕾,兰天维,黎扬辉,等.影响红掌愈伤组织诱导、增殖和芽分化的因素[J].种子,2006,25(11):26-30.
[24] 李千,孙丹,李宏博,等.北五味子愈伤组织增殖及胚性愈伤组织诱导的研究[J].种子,2014,33(6):5-9.
[25] 王新,成仿云,钟原,等.凤丹牡丹鳞芽离体培养与快繁技术[J].林业科学,2016,52(5):101-110.
[26] DAS J,MAO A A,HANDIQUE P J.Callus-mediated organogenesis and effect of growth regulators on production of different valepotriates in Indian valerian (Valeriana jatamansi Jones.)[J].Acta Physiologiae Plantarum,2013,35(1):55-63.
[27] 何朋,刘思妤,王宏宇,等.北细辛叶柄愈伤组织的增殖培养及器官分化研究[J].中草药,2016,47(13):2341-2345.
[28] 赵金梅,李芳,周禾,等.紫花苜蓿组织培养体系的建立[J].核农学报,2010,24(3):507-512.