植物生长调节剂优化不同品种冬小麦成熟胚再生体系
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摘要
小麦愈伤组织再生能力是遗传转化的重要基础。为优化小麦成熟胚再生体系,以科农199、轮选987和济麦22三个冬小麦品种的成熟胚为外植体,在愈伤组织分化阶段添加不同浓度的植物生长调节剂TDZ(N-苯基-N′-1, 2, 3-噻二唑-5-脲,thidiazuron),统计和分析分化率。结果表明,TDZ最适响应浓度存在品种间差异,科农199、轮选987和济麦22成熟胚愈伤组织分化的最适TDZ浓度分别为1.5、1.0、0.5 mg/L,愈伤组织分化率分别为84.61%、71.88%和71.77%,均极显著高于对照。本研究结果可为优化冬小麦成熟胚愈伤组织再生体系提供参考。
Wheat callus regeneration ability is an important basis of genetic transformation. To optimize wheat mature embryo regeneration system, the mature embryos of three winter wheat varieties, Kenong 199, Lunxuan 987 and Jimai 22, were used as explants. Different concentrations of plant growth regulator TDZ(thidiazuron) were added into the culture medium in callus differentiation stage, and the regeneration efficiency of wheat mature embryo was calculated and analyzed. The results showed that the optimal TDZ concentration for callus differentiation of Kenong 199, Lunxuan 987 and Jimai 22 was 1.5, 1.0 mg/L and 0.5 mg/L respectively, and the callus regeneration efficiency was 84.61%, 71.88% and 71.77%, respectively, which were significantly higher than those of the control. This results provided a reference for improving callus regeneration ability of winter wheat mature embryos.
Wheat callus regeneration ability is an important basis of genetic transformation. To optimize wheat mature embryo regeneration system, the mature embryos of three winter wheat varieties, Kenong 199, Lunxuan 987 and Jimai 22, were used as explants. Different concentrations of plant growth regulator TDZ(thidiazuron) were added into the culture medium in callus differentiation stage, and the regeneration efficiency of wheat mature embryo was calculated and analyzed. The results showed that the optimal TDZ concentration for callus differentiation of Kenong 199, Lunxuan 987 and Jimai 22 was 1.5, 1.0 mg/L and 0.5 mg/L respectively, and the callus regeneration efficiency was 84.61%, 71.88% and 71.77%, respectively, which were significantly higher than those of the control. This results provided a reference for improving callus regeneration ability of winter wheat mature embryos.
引文
[1] 叶兴国,徐惠君,赵乐莲,等.组织培养途径改良定型小麦品种的研究[J].作物学报,1998,24(3):310-314.
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[16] Yu H,Wang W,Wang Y,et al.High frequency wheat regeneration from leaf tissue explants of regenerated plantlets[J].Advances in Bioscience and Biotechnology,2012,3(1):46-50.
[17] 王丽,陈耀锋,张月琴,等.有机添加物对小麦成熟胚愈伤组织分化特性的影响[J].西北农业学报,2014,23(3):45-49.
[18] Thomas J,Katterman F.Cytokinin activity induced by thidiazuron[J].Plant Physiology,1986,81(2):681-683.
[19] Murthy B,Saxena P.Somatic embryogenesis and plant regeneration of neem (Azadirachta indica A.Juss.)[J].Plant Cell Reports,1998,17(6/7):469-475.
[20] Murch S,Saxena P.The role of proline in thidiazuron induced somatic embryogenesis of peanut[J].In Vitro Cellular & Developmental Biology-Plant,1999,35(1):102-105.
[21] Mok M,Mok D,Armsstrong D.Cytokin activity of N-phenye-N′-1,2,3-thidiazol-5-ylurea (thidiazuron)[J].Phytochemistry,1982,21(7):1509-1511.
[22] Hutchinson M,Saxena P.Role of purine metabolism in thidiazuron-induced somatic embryogenesis of geranium (Pelargonium× hortorum) hypocotyl cultures[J].Physiologia Plantarum,1996,98(3):517-522.
[23] 徐华松,徐九龙.TDZ在植物组织培养中的作用[J].广西植物,1996,16(1):77-80.
[24] 周俊彦,郭扶兴.苯基脲衍生物的细胞分裂素活性[J].植物生理学通讯,1990(4):7-13.
[2] 武丽敏,郑有良,魏育明,等.小麦转基因研究进展[J].四川农业大学学报,2003,21(2):176-181.
[3] Fennell S,Bohorova N,Ginkel M,et al.Plant regeneration from immature embryos of 48 elite CIMMYT bread wheats[J].Theoretical and Applid Genetics,1996,92(2):163-169.
[4] Mamrutha H,Kumar R,Venkatesh K,et al.Genetic transformation of wheat-present status and future potential[J].Journal of Wheat Research,2014,6(2):107-119.
[5] Carman J G,Jefferson N E,Campbell W F.Induction of embryogenic Triticum aestivum L.calli.I.Quantification of genotype and culture medium effects[J].Plant Cell,Tissue and Organ Culture,1987,10:101-103.
[6] Ramage C,Williams R.Mineral nutrition and plant morphogenesis[J].In Vitro Cellular & Developmental Biology-Plant,2002,38(2):116-124.
[7] 别晓敏,杜丽璞,徐惠君,等.培养基中CuSO4和Fe盐浓度对小麦胚培养再生效果的影响[J].植物遗传资源学报,2011,12(6):975-981.
[8] 陈云凤,张春荣,黄霞,等.TDZ对植物体细胞胚胎发生的作用[J] .植物生理学通讯,2006,42(1):127-133.
[9] Huetteman C,Preece J.Thidiazuron:a potent cytokinin for woody plant tissue culture[J].Plant Cell,Tissue and Organ Culture,1993,33(2):105-119.
[10] Loschiavo F,Pitto L,Giulilano G.DNA methylation of embryogenic carrot cell cultures and its variation as caused by mutation,differentiation,hormones and hypomethylating drugs[J].Theoretical and Applied Genetics,1989,77(3):325-331.
[11] 徐晓峰,黄学林.TDZ:一种有效的植物生长调节剂[J].植物学通报,2003,20(2):227-237.
[12] Hakam N,Udupa S,Rabha A,et al.Efficient callus induction and plantlets regeneration in bread wheat using immature and mature embryos[J].International Journal of Biotechnology Research,2015,3(1):1-9.
[13] 郭志江,张凤路,王金明,等.农杆菌介导小麦成熟胚体系的优化[J].中国农学通报,2008,24(4):181-185.
[14] 陶丽莉,殷桂香,叶兴国.小麦成熟胚组织培养及遗传转化研究进展[J].麦类作物学报,2008,28(4):713-718.
[15] Shariatpanahi M,Belogradova K,Hessamvaziri L,et al.Efficient embryogenesis and regeneration in freshly isolated and cultured wheat (Triticum aestivum L.) microspores without stress pretreatment[J].Plant Cell Reports,2006,25(12):1294-1299.
[16] Yu H,Wang W,Wang Y,et al.High frequency wheat regeneration from leaf tissue explants of regenerated plantlets[J].Advances in Bioscience and Biotechnology,2012,3(1):46-50.
[17] 王丽,陈耀锋,张月琴,等.有机添加物对小麦成熟胚愈伤组织分化特性的影响[J].西北农业学报,2014,23(3):45-49.
[18] Thomas J,Katterman F.Cytokinin activity induced by thidiazuron[J].Plant Physiology,1986,81(2):681-683.
[19] Murthy B,Saxena P.Somatic embryogenesis and plant regeneration of neem (Azadirachta indica A.Juss.)[J].Plant Cell Reports,1998,17(6/7):469-475.
[20] Murch S,Saxena P.The role of proline in thidiazuron induced somatic embryogenesis of peanut[J].In Vitro Cellular & Developmental Biology-Plant,1999,35(1):102-105.
[21] Mok M,Mok D,Armsstrong D.Cytokin activity of N-phenye-N′-1,2,3-thidiazol-5-ylurea (thidiazuron)[J].Phytochemistry,1982,21(7):1509-1511.
[22] Hutchinson M,Saxena P.Role of purine metabolism in thidiazuron-induced somatic embryogenesis of geranium (Pelargonium× hortorum) hypocotyl cultures[J].Physiologia Plantarum,1996,98(3):517-522.
[23] 徐华松,徐九龙.TDZ在植物组织培养中的作用[J].广西植物,1996,16(1):77-80.
[24] 周俊彦,郭扶兴.苯基脲衍生物的细胞分裂素活性[J].植物生理学通讯,1990(4):7-13.