百脉根愈伤组织诱导研究
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摘要
以MS为基本培养基,利用正交试验设计,研究了外植体类型、外源激素及其浓度配比对百脉根愈伤组织诱导的影响。结果表明:外植体类型和外源激素对百脉根愈伤组织诱导和褐化的影响均达到极显著水平(P<0.01),其中2,4-D浓度是影响百脉根愈伤组织诱导的主要因子,而NAA浓度是次要因子,以下胚轴为最佳外植体,添加1.0~2.0mg/L 2,4-D、0.5mg/L KT、0.5mg/L 6-BA进行愈伤组织培养效果最佳,诱导率可达100%。
Using MS as the basic culture medium and orthogonal test to study the influence of Lotus corniculatus L.callus induction of two factors that one is exogenous hormone concentration,the other is explants type,screening efficient Lotus corniculatus L.callus induction system.The results showed that:five factors(explants type,2,4-D,KT,NAA,6-BA)on Lotus corniculatus L.callus induction rate and browning rate effects were achieved significant level(P<0.01).The main factors affecting on Lotus corniculatus L.callus induction rate was the concentration of 2,4-D,secondly was the types of explants,the most helpful explants was hypocotyls;the best effect culture medium of callus induction was using hypocotyls as explants and adding 1-2.0mg/L 2,4-D,0.5mg/L KT,0.5mg/L 6-BA within MS,the induction rate was 100%.the most influential factor on the browning rate was the concentration of NAA,followed by explants types,KT,6-BA,with the least influence was the concentration of 2,4-D.
Using MS as the basic culture medium and orthogonal test to study the influence of Lotus corniculatus L.callus induction of two factors that one is exogenous hormone concentration,the other is explants type,screening efficient Lotus corniculatus L.callus induction system.The results showed that:five factors(explants type,2,4-D,KT,NAA,6-BA)on Lotus corniculatus L.callus induction rate and browning rate effects were achieved significant level(P<0.01).The main factors affecting on Lotus corniculatus L.callus induction rate was the concentration of 2,4-D,secondly was the types of explants,the most helpful explants was hypocotyls;the best effect culture medium of callus induction was using hypocotyls as explants and adding 1-2.0mg/L 2,4-D,0.5mg/L KT,0.5mg/L 6-BA within MS,the induction rate was 100%.the most influential factor on the browning rate was the concentration of NAA,followed by explants types,KT,6-BA,with the least influence was the concentration of 2,4-D.
引文
[1]杜桂娟,马凤江,刘晓宏,等.优良园林绿化地被植物百脉根[J].北方园艺,2006,(2):107.
[2]孙艳香,杨红梅,耿云红,等.根癌农杆菌介导的百脉根遗传转化体系的优化研究[J].南开大学学报(自然科学版),2006,(02):51-57.
[3]俞小秋,丁玉川,白增森,等.优良牧草——百脉根[J].北京农业,2000,(7):29-30.
[4]Atika C,Deepak P.Regeneration and genetic transformation of grain legumes:An overview[J].Current Science,2003,(84):3381-387.
[5]Susana de S A,Ana S R L A,DulceMMFS,et al.An efficient transformation method to regenerate a high number of transgenic plants using a new embryogenic line of Medicago truncatula cv.Jemalong[J].Plant Cell,Tissue and Organ Culture,2004,(78):123-131.
[6]Handberg K,and Stougaard.Lotus japonicus,an autogamous,diploid legume species for classical and molecular genetics[J].Plant J,1992,2(4):487-496.
[7]韩阳,卢福荣,崔智昕,等.百脉根植株高频再生体系的建立[J].辽宁大学学报(自然科学版),2014,(02):152-156.
[8]Aoki T,Kamizawa A,and Ayabe S.Efficient Agrobacterium mediated transformation of Lotus japonicus with reliable antibiotic selection[J].Plant Cell Rep,2002,21(3):238-243.
[9]贾永红,孙艳香,张江丽,等.百脉根高频再生体系建立和猪瘟病毒E2基因的遗传转化[J].湖北农业科学,2014,(10):2368-2372.
[10]宋莉,赵德刚.百脉根离体再生体系的优化[J].分子植物育种,2015,13(4):910-914.
[11]李春喜.生物统计学(第一版)[M].北京:科学出版社,1997.198-204.
[12]池慧芳.豆科植物组织培养研究进展[J].内蒙古农业科技,2014,(3):15-16.
[13]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.
[14]伊风艳,石凤翎,展春芳,姜圆圆.黄花苜蓿愈伤组织诱导及分化培养条件的研究[J].中国草地学报,2011,(06):14-20.
[15]王涌鑫,关宁,李聪.高效的苜蓿组织培养再生体系的建立[J].东北师大学报(自然科学版),2008,40(3):112-117.
[16]李芳,赵金梅,石凤翎,等.影响中苜2号苜蓿愈伤组织诱导的主要因素分析[J].中国草地学报,2009,(04):47-52.
[17]史毅,马晖玲.三个苜蓿品种植株高频再生体系的研究[J].甘肃农业大学学报,2014,(02):125-132.
[18]李娟,张万军,王涛.黄花苜蓿高频植株再生体系建立的研究[J].草地学报,2014,(04):834-839.
[19]张万军,王涛.紫花苜蓿愈伤成苗高频再生体系的建立及其影响因子的研究[J].中国农业科学,2002,35(12):1579-1583.
[20]马晖玲,卢欣石,曹致中,等.紫花苜蓿不同栽培品种植株再生的研究[J].草业学报,2004,13(6):99-105.
[21]唐广立.百脉根高频再生体系的建立及兔出血症病毒衣壳蛋白VP60基因的转化[D].西北农林科技大学,2007.
[22]曾镭,刘燕.植物组织培养中褐化问题的研究进展[J].安徽农学通报,2007,13(14):49-50.
[2]孙艳香,杨红梅,耿云红,等.根癌农杆菌介导的百脉根遗传转化体系的优化研究[J].南开大学学报(自然科学版),2006,(02):51-57.
[3]俞小秋,丁玉川,白增森,等.优良牧草——百脉根[J].北京农业,2000,(7):29-30.
[4]Atika C,Deepak P.Regeneration and genetic transformation of grain legumes:An overview[J].Current Science,2003,(84):3381-387.
[5]Susana de S A,Ana S R L A,DulceMMFS,et al.An efficient transformation method to regenerate a high number of transgenic plants using a new embryogenic line of Medicago truncatula cv.Jemalong[J].Plant Cell,Tissue and Organ Culture,2004,(78):123-131.
[6]Handberg K,and Stougaard.Lotus japonicus,an autogamous,diploid legume species for classical and molecular genetics[J].Plant J,1992,2(4):487-496.
[7]韩阳,卢福荣,崔智昕,等.百脉根植株高频再生体系的建立[J].辽宁大学学报(自然科学版),2014,(02):152-156.
[8]Aoki T,Kamizawa A,and Ayabe S.Efficient Agrobacterium mediated transformation of Lotus japonicus with reliable antibiotic selection[J].Plant Cell Rep,2002,21(3):238-243.
[9]贾永红,孙艳香,张江丽,等.百脉根高频再生体系建立和猪瘟病毒E2基因的遗传转化[J].湖北农业科学,2014,(10):2368-2372.
[10]宋莉,赵德刚.百脉根离体再生体系的优化[J].分子植物育种,2015,13(4):910-914.
[11]李春喜.生物统计学(第一版)[M].北京:科学出版社,1997.198-204.
[12]池慧芳.豆科植物组织培养研究进展[J].内蒙古农业科技,2014,(3):15-16.
[13]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.
[14]伊风艳,石凤翎,展春芳,姜圆圆.黄花苜蓿愈伤组织诱导及分化培养条件的研究[J].中国草地学报,2011,(06):14-20.
[15]王涌鑫,关宁,李聪.高效的苜蓿组织培养再生体系的建立[J].东北师大学报(自然科学版),2008,40(3):112-117.
[16]李芳,赵金梅,石凤翎,等.影响中苜2号苜蓿愈伤组织诱导的主要因素分析[J].中国草地学报,2009,(04):47-52.
[17]史毅,马晖玲.三个苜蓿品种植株高频再生体系的研究[J].甘肃农业大学学报,2014,(02):125-132.
[18]李娟,张万军,王涛.黄花苜蓿高频植株再生体系建立的研究[J].草地学报,2014,(04):834-839.
[19]张万军,王涛.紫花苜蓿愈伤成苗高频再生体系的建立及其影响因子的研究[J].中国农业科学,2002,35(12):1579-1583.
[20]马晖玲,卢欣石,曹致中,等.紫花苜蓿不同栽培品种植株再生的研究[J].草业学报,2004,13(6):99-105.
[21]唐广立.百脉根高频再生体系的建立及兔出血症病毒衣壳蛋白VP60基因的转化[D].西北农林科技大学,2007.
[22]曾镭,刘燕.植物组织培养中褐化问题的研究进展[J].安徽农学通报,2007,13(14):49-50.