豌豆离体再生体系建立及遗传稳定性研究
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摘要
【目的】提高速豌豆诱伤组织和不定芽的诱导率,建立豌豆高效离体再生体系.【方法】以半无叶型豌豆品种‘陇豌1号’和蔓生型豌豆品种‘S3008’的茎段为外植体,研究不同基因型、培养基激素配比对愈伤组织诱导、分化的影响.【结果】‘陇豌1号’茎段愈伤组织诱导率和分化率较高,分别达到88.7%和76.7%,‘S3008’茎段最高愈伤组织诱导率和分化率分别为86.7%和74.7%.诱导愈伤组织最适培养基是MS+2mg/L TDZ+0.2mg/L 2,4-D;诱导不定芽形成的最适培养基为MS+2mg/L TDZ+1mg/L 6-BA,平均不定芽数为4.3;在不定芽诱导生根培养基(1/2MS+20g/L蔗糖+1.5mg/L ABT)上,生根率为74.0%,移栽后再生植株成活率达86.0%.两个豌豆品种都能得到再生植株且繁殖系数高.【结论】本培养体系适宜豌豆离体再生.通过染色体分析,再生植株染色体数和供体亲本材料一致,初步表明再生植株遗传稳定.
【Objective】To increase the rate of callus inductive and adventitious bud induction,and to set up new regeneration system.【Method】The stems of semi-leafless peas cultivars‘Longwan NO.1'and vining peas cultivar‘S3008'were used as explants,the effect of different genotypes,hormone types and the ratio of medium were studied on callus induction,callus differentiation and rooting.【Result】The higher frequency of callus initiation and differentiation from stems of‘Longwan NO.1'was 88.7% and 76.7%.The frequency of callus initiation and differentiation from stems of‘S3008'was 86.7% and 74.7%.The optimal medium for callus induction was MS+2mg/LTDZ+0.2mg/L 2,4-D.The appropriate medium for adventitious bud induction was MS+2 mg/L TDZ+1 mg/L 6-BA,the adventitious bud number per explants was 4.3.The adventitious bud on rooting induction medium were 1/2MS+20 g/L sucrose+1.5mg/L ABT,the rooting rates was 74% and the survival rates of regenerated plants were 86%.【Conclusion】The two cultivars tested are all able to produce regeneration plant and propagate coefficient is high,shows that this new regeneration system could be applicable to regeneration of peas in vitro.By chro-mosome analysis,chromosome number of the regenerated plants are same to the donor plants,which is suggested that the regenerated plants are genetic stable.
【Objective】To increase the rate of callus inductive and adventitious bud induction,and to set up new regeneration system.【Method】The stems of semi-leafless peas cultivars‘Longwan NO.1'and vining peas cultivar‘S3008'were used as explants,the effect of different genotypes,hormone types and the ratio of medium were studied on callus induction,callus differentiation and rooting.【Result】The higher frequency of callus initiation and differentiation from stems of‘Longwan NO.1'was 88.7% and 76.7%.The frequency of callus initiation and differentiation from stems of‘S3008'was 86.7% and 74.7%.The optimal medium for callus induction was MS+2mg/LTDZ+0.2mg/L 2,4-D.The appropriate medium for adventitious bud induction was MS+2 mg/L TDZ+1 mg/L 6-BA,the adventitious bud number per explants was 4.3.The adventitious bud on rooting induction medium were 1/2MS+20 g/L sucrose+1.5mg/L ABT,the rooting rates was 74% and the survival rates of regenerated plants were 86%.【Conclusion】The two cultivars tested are all able to produce regeneration plant and propagate coefficient is high,shows that this new regeneration system could be applicable to regeneration of peas in vitro.By chro-mosome analysis,chromosome number of the regenerated plants are same to the donor plants,which is suggested that the regenerated plants are genetic stable.
引文
[1]郑卓杰,王述民,宗绪晓.中国食用豆类学[M].北京:中国农业出版社,1997
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[4]Schroeder H E,Schotz A H,Wardley-Richardson T,et al.Transformation and regeneration of two cultivars of pea(Pisum sativum L.)[J].Plant Physiology,1993,101(3):751-757
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[6]苏承刚,吴学科,郑占伟,等.食荚型豌豆组织培养和植株再生研究[J].西南师范大学学报:自然科学版,2007,32(4):30-32
[7]杨起简,周禾,孙彦,等.豌豆染色体制片技术的比较研究[J].北京农学院学报,2003,18(3):172-173
[8]高金辉,王玲,张厚良,等.山葡萄组织培养方法[J].东北林业大学学报,2007,35(7):37-39
[9]张晨,王巍,赵溦,等.山野豌豆组织培养及无性系建立的研究[J].黑龙江科学,2012,3(3):8-11
[10]Tzitzikas E N,Bergervoet M,Raemakers K,et al.Regeneration of pea(Pisum sativum L.)by a cyclic organogenic system[J].Plant cell Reports,2004,23(7):453-460
[11]那光宇,张姝媛,郭娜,等.什锦丁香花芽分化过程中植物内源激素的变化[J].内蒙古农业大学学报:自然科学版,2012,Z1:58-61
[12]Radke S E,Turner J C,Facciotti D.Transformation and regeneration of Brassica rapausing Agrobacterium tumefaciens[J].Plant Cell Reports,1992,11(10):499-505
[13]马光,周波,李玉花.芜菁高频率再生体系的建立及优化[J].园艺学报,2008,35(6):833-840
[14]Khalafalla M M,Hattori K.Differential in vitro direct shoot regeneration responses in embryo axis and shoot tip explants of faba bean[J].Breeding Science,2000,50(2):117-122
[15]吴琼.油莎豆再生体系的建立及多倍体诱导[D].兰州:兰州大学,2009
[2]杨晓明,任瑞玉.国内外豌豆生产和育种研究进展[J].甘肃农业科技,2005(8):3-5
[3]罗志娜,赵桂琴,刘欢.燕麦成熟胚的组织培养及植株再生[J].甘肃农业大学学报,2012,47:60-68
[4]Schroeder H E,Schotz A H,Wardley-Richardson T,et al.Transformation and regeneration of two cultivars of pea(Pisum sativum L.)[J].Plant Physiology,1993,101(3):751-757
[5]Bean S J,Gooding P S,Mullincaux P M,et al.A simple system for pea transformation[J].Plant Cell Reports,1997,16(8):513-519
[6]苏承刚,吴学科,郑占伟,等.食荚型豌豆组织培养和植株再生研究[J].西南师范大学学报:自然科学版,2007,32(4):30-32
[7]杨起简,周禾,孙彦,等.豌豆染色体制片技术的比较研究[J].北京农学院学报,2003,18(3):172-173
[8]高金辉,王玲,张厚良,等.山葡萄组织培养方法[J].东北林业大学学报,2007,35(7):37-39
[9]张晨,王巍,赵溦,等.山野豌豆组织培养及无性系建立的研究[J].黑龙江科学,2012,3(3):8-11
[10]Tzitzikas E N,Bergervoet M,Raemakers K,et al.Regeneration of pea(Pisum sativum L.)by a cyclic organogenic system[J].Plant cell Reports,2004,23(7):453-460
[11]那光宇,张姝媛,郭娜,等.什锦丁香花芽分化过程中植物内源激素的变化[J].内蒙古农业大学学报:自然科学版,2012,Z1:58-61
[12]Radke S E,Turner J C,Facciotti D.Transformation and regeneration of Brassica rapausing Agrobacterium tumefaciens[J].Plant Cell Reports,1992,11(10):499-505
[13]马光,周波,李玉花.芜菁高频率再生体系的建立及优化[J].园艺学报,2008,35(6):833-840
[14]Khalafalla M M,Hattori K.Differential in vitro direct shoot regeneration responses in embryo axis and shoot tip explants of faba bean[J].Breeding Science,2000,50(2):117-122
[15]吴琼.油莎豆再生体系的建立及多倍体诱导[D].兰州:兰州大学,2009