小滨菊组培快繁体系的初步建立
|
摘要
为大量扩繁小滨菊幼苗,离体保存核心种质,进而创新菊花优异种质,培育耐涝菊花新优品种,解决菊花忌涝问题,以多年生沼生植物小滨菊(Leucanthemella linearis)的带芽茎段为外植体,开展小滨菊组培快繁体系建立的研究。结果表明,小滨菊茎段的最佳启动培养基为MS+0.5 mg/L 6-BA+0.5 mg/LNAA,启动率达到80%;最佳继代培养基为MS+0.5 mg/L 6-BA+0.2 mg/LNAA,增殖效果良好,出芽指数平均达到9.2;最佳生根培养基为1/2MS+0.3 mg/L NAA,生根率85.72%,根系生长状态良好。小滨菊茎段诱导愈伤组织的最佳培养基为MS+0.2 mg/L 6-BA+0.5 mg/L NAA,愈伤组织分化率最高的培养基为MS+1.0 mg/L 6-BA+0.2 mg/L NAA,分化率可达70.00%。
The paper aims to propagate and conserve the key germplasms and innovate the elite germplasm of Chrysanthemums used for breeding new excellent cultivars. Experiments were carried out for the standardization of micropropagation and regeneration of Leucanthemella linearis, a helophyte perennial plant.Nodal segments were used as explants and were cultured on MS medium supplemented with different concentrations of 6-BA and NAA for shoot inductions and the best result of primary cultural medium obtained was MS medium supplemented with 6-BA and NAA at the concentration of 0.5 mg/L, respectively. The best subcultural medium found was MS medium supplemented with 6-BA+NAA at the concentration of 0.5 mg/L and 0.2 mg/L, respectively, resulting about 9.2 multiple shoots from a single shoot tip explant. For root induction and elongation, 1/2 MS medium plus NAA at the concentration of 0.3 mg/Lshowed good effect, and the rooting percentage was 85.72%. Callus was induced from nodal segment on MS medium supplemented with0.2 mg/L 6-BA and 0.5 mg/L NAA, and plants regenerated best from these calluses on MS medium supplemented with 1.0 mg/L 6-BA and 0.2 mg/L NAA, the ratio of regeneration was 70.00%.
The paper aims to propagate and conserve the key germplasms and innovate the elite germplasm of Chrysanthemums used for breeding new excellent cultivars. Experiments were carried out for the standardization of micropropagation and regeneration of Leucanthemella linearis, a helophyte perennial plant.Nodal segments were used as explants and were cultured on MS medium supplemented with different concentrations of 6-BA and NAA for shoot inductions and the best result of primary cultural medium obtained was MS medium supplemented with 6-BA and NAA at the concentration of 0.5 mg/L, respectively. The best subcultural medium found was MS medium supplemented with 6-BA+NAA at the concentration of 0.5 mg/L and 0.2 mg/L, respectively, resulting about 9.2 multiple shoots from a single shoot tip explant. For root induction and elongation, 1/2 MS medium plus NAA at the concentration of 0.3 mg/Lshowed good effect, and the rooting percentage was 85.72%. Callus was induced from nodal segment on MS medium supplemented with0.2 mg/L 6-BA and 0.5 mg/L NAA, and plants regenerated best from these calluses on MS medium supplemented with 1.0 mg/L 6-BA and 0.2 mg/L NAA, the ratio of regeneration was 70.00%.
引文
[1]尹冬梅,管志勇,陈素梅,等.菊花及其近缘种属植物耐涝性评价体系建立及耐涝性鉴定[J].植物遗传资源学报,2009,10(3):399-404.
[2] Tanaka R, Shimotomai N. Cytogenetic studies on the F1hybrid ofChrysanthemum lineare(now Leucanthemella lineare)Ch.nipponicum(now Nipponanthemum nipponicum)[J]. Zeitschrift FürVererbungslehre, 1961,92:190-196.
[3] Zhao H B, Chen F D, Chen S M, et al. Molecular phylgeny ofChrysanthemum, Ajania and its allies(Anthemideae, Asteraceae)asinferred from nuclear ribosomal ITS and chloroplast trnL-F IGSsequences[J]. Plant Systematics and Evolution,2010,284(3):153-169.
[4] Hisakazu Ogura, Katsuhiko Kondo. Application of genomic in situhybridization to the chromosome complement of the intergenerichybrid between Leucanthemella linearis(Matsum.)Tzuvelev andNipponanthemum nipponicum(Franch. Et Maxim.)Kitamura[J].Chromosome Science,1998,2:91-93.
[5] Hong G, Wu X B, Liu Y C, et al. Intergeneric hybridizationbetween Hippolytia kaschgarica(Krascheninnikov)Poljakov andNipponanthemum nipponicum(Franch. Ex Maxim.)Kitam[J].Genetic Resourced&Crop Evolution,2015,62(2):255-263.
[6] Magdy Hussein, Abd El-Twab, Katsuhiko Kondo. Identification ofparental chromosomes and changes of artificial, intergeneric F1hybrid between Dendranthema horaimontana and Nipponanthemumnipponicum by fluorescence genomic in situ hybridization(GISH)and fluorescence in situ hybridization(FISH)[J]. ChromosomeScience,2004,8(3):71-79.
[7] Jaime A, Teixeira da Silva, Justyna Lema-Ruminska, et al.Regeneration from chrusanthemum flowers:a review[J]. ActaPhysiologiae Plantarum,2015,37(2):1-16.
[8]程越.菊花脑再生体系建立的研究[D].南京:南京农业大学,2014.
[9]谢菲,浦娅,赵惠恩.北极菊组培快繁体系的建立[A].中国观赏园艺研究进展[M].2015:260-263
[10]刘彬.太行菊组织培养及其快速繁育技术研究[D].郑州:郑州大学,2016.
[11]杨德艳,赵惠恩.戈壁短舌菊的组织培养与快速繁殖[J].植物生理学通讯,2010,46(3):255-256.
[12]符运柳,王永壮,郭庆辉,等.菊花脑的组织培养与快速繁殖[J].安徽农业科学,2010,38(24):13367-13368.
[13]董春艳.神农香菊(Dendranthema indicum var. aromaticum)组织培养及细胞悬浮培养体系的建立[D].哈尔滨:东北林业大学,2011.
[14] Segio Echeverrigaray, Simone Biaso, Fernando Fracaro, et al.Clonal Micropropagation of Roman Chamomile(Anthemis nobilisL.)[J]. Journal of Herbs, Spices&Medicinal Plants,2000,7(2):35-41.
[15]郑燕,沈景,韩倩,等.紫花亚菊的组织培养与快速繁殖[J].植物生理学报,2011,47(10):983-986.
[16] Jaime A, Teixeira da Silva. Anthemideae:advances in tissue culture,genetics and transgenic biotechnology[J]. African Journal ofBiotechnology,2003,2(12):547-556.
[17] Chang Hee Lee, Ki Sun Kim. Micropropagation of Dendranthemazawadaskii Tzvelev Complex Native to Korea[J]. HorticultureEnvironment Biotechnol,2010,51(2):115-122.
[18]郑成淑,孙宪芝,王文莉,等.光周期对菊花花芽分化及其叶片和芽内源多胺含量的影响[J].西北植物学报,2008(7):1349-1353.
[19]姚洪军,罗晓芳,田砚亭.植物组织培养外植体褐变的研究进展[J].北京林业大学学报,1999(3):81-87.
[20]徐振彪,傅作申,原亚萍,等.植物组织培养过程中的褐化现象[J].国外农学:杂粮作物,1997(1):56-57.
[21]金顺福,姜成模,崔哲官,等.培育健壮马铃薯试管苗试验[J].马铃薯杂志,1995,9(3):139-143.
[22]郎贤波,廉美兰,朴炫春,等.无机盐浓度对马铃薯脱毒苗微繁的影响[J].延边大学农学学报,2007,29(1):1-4
[23]常立国,范惠玲,刘建超,等.马铃薯试管苗壮苗和生根培养影响因素的研究[J].作物杂志,2016(2):129-132
[24]赵光磊,吴凌娟,张雅奎,等.马铃薯脱毒试管苗壮苗培养体系的优化[J].中国马铃薯,2012,26(4):200-205
[25]李露华,潘超美,苏家贤,等.贡菊组培苗壮苗生根的研究[J].广州中医药大学学报,2015,32(3):494-498.
[26]梁芳,蒋素华,王洁琼,等.濒危植物太行菊组织培养及快繁技术研究[J].中国农学通报,2015,31(16):115-120.
[27]姚连芳,董美华,毛玉收.太行菊组织培养研究[J].中国农学通报,2004,20(6):29-31.
[28]王建博,徐思明,董鹏,等.太行菊顶芽离体高效再生系的建立[J].首都师范大学学报:自然科学版,2008,29(5):45-50.
[2] Tanaka R, Shimotomai N. Cytogenetic studies on the F1hybrid ofChrysanthemum lineare(now Leucanthemella lineare)Ch.nipponicum(now Nipponanthemum nipponicum)[J]. Zeitschrift FürVererbungslehre, 1961,92:190-196.
[3] Zhao H B, Chen F D, Chen S M, et al. Molecular phylgeny ofChrysanthemum, Ajania and its allies(Anthemideae, Asteraceae)asinferred from nuclear ribosomal ITS and chloroplast trnL-F IGSsequences[J]. Plant Systematics and Evolution,2010,284(3):153-169.
[4] Hisakazu Ogura, Katsuhiko Kondo. Application of genomic in situhybridization to the chromosome complement of the intergenerichybrid between Leucanthemella linearis(Matsum.)Tzuvelev andNipponanthemum nipponicum(Franch. Et Maxim.)Kitamura[J].Chromosome Science,1998,2:91-93.
[5] Hong G, Wu X B, Liu Y C, et al. Intergeneric hybridizationbetween Hippolytia kaschgarica(Krascheninnikov)Poljakov andNipponanthemum nipponicum(Franch. Ex Maxim.)Kitam[J].Genetic Resourced&Crop Evolution,2015,62(2):255-263.
[6] Magdy Hussein, Abd El-Twab, Katsuhiko Kondo. Identification ofparental chromosomes and changes of artificial, intergeneric F1hybrid between Dendranthema horaimontana and Nipponanthemumnipponicum by fluorescence genomic in situ hybridization(GISH)and fluorescence in situ hybridization(FISH)[J]. ChromosomeScience,2004,8(3):71-79.
[7] Jaime A, Teixeira da Silva, Justyna Lema-Ruminska, et al.Regeneration from chrusanthemum flowers:a review[J]. ActaPhysiologiae Plantarum,2015,37(2):1-16.
[8]程越.菊花脑再生体系建立的研究[D].南京:南京农业大学,2014.
[9]谢菲,浦娅,赵惠恩.北极菊组培快繁体系的建立[A].中国观赏园艺研究进展[M].2015:260-263
[10]刘彬.太行菊组织培养及其快速繁育技术研究[D].郑州:郑州大学,2016.
[11]杨德艳,赵惠恩.戈壁短舌菊的组织培养与快速繁殖[J].植物生理学通讯,2010,46(3):255-256.
[12]符运柳,王永壮,郭庆辉,等.菊花脑的组织培养与快速繁殖[J].安徽农业科学,2010,38(24):13367-13368.
[13]董春艳.神农香菊(Dendranthema indicum var. aromaticum)组织培养及细胞悬浮培养体系的建立[D].哈尔滨:东北林业大学,2011.
[14] Segio Echeverrigaray, Simone Biaso, Fernando Fracaro, et al.Clonal Micropropagation of Roman Chamomile(Anthemis nobilisL.)[J]. Journal of Herbs, Spices&Medicinal Plants,2000,7(2):35-41.
[15]郑燕,沈景,韩倩,等.紫花亚菊的组织培养与快速繁殖[J].植物生理学报,2011,47(10):983-986.
[16] Jaime A, Teixeira da Silva. Anthemideae:advances in tissue culture,genetics and transgenic biotechnology[J]. African Journal ofBiotechnology,2003,2(12):547-556.
[17] Chang Hee Lee, Ki Sun Kim. Micropropagation of Dendranthemazawadaskii Tzvelev Complex Native to Korea[J]. HorticultureEnvironment Biotechnol,2010,51(2):115-122.
[18]郑成淑,孙宪芝,王文莉,等.光周期对菊花花芽分化及其叶片和芽内源多胺含量的影响[J].西北植物学报,2008(7):1349-1353.
[19]姚洪军,罗晓芳,田砚亭.植物组织培养外植体褐变的研究进展[J].北京林业大学学报,1999(3):81-87.
[20]徐振彪,傅作申,原亚萍,等.植物组织培养过程中的褐化现象[J].国外农学:杂粮作物,1997(1):56-57.
[21]金顺福,姜成模,崔哲官,等.培育健壮马铃薯试管苗试验[J].马铃薯杂志,1995,9(3):139-143.
[22]郎贤波,廉美兰,朴炫春,等.无机盐浓度对马铃薯脱毒苗微繁的影响[J].延边大学农学学报,2007,29(1):1-4
[23]常立国,范惠玲,刘建超,等.马铃薯试管苗壮苗和生根培养影响因素的研究[J].作物杂志,2016(2):129-132
[24]赵光磊,吴凌娟,张雅奎,等.马铃薯脱毒试管苗壮苗培养体系的优化[J].中国马铃薯,2012,26(4):200-205
[25]李露华,潘超美,苏家贤,等.贡菊组培苗壮苗生根的研究[J].广州中医药大学学报,2015,32(3):494-498.
[26]梁芳,蒋素华,王洁琼,等.濒危植物太行菊组织培养及快繁技术研究[J].中国农学通报,2015,31(16):115-120.
[27]姚连芳,董美华,毛玉收.太行菊组织培养研究[J].中国农学通报,2004,20(6):29-31.
[28]王建博,徐思明,董鹏,等.太行菊顶芽离体高效再生系的建立[J].首都师范大学学报:自然科学版,2008,29(5):45-50.