铁皮石斛2n花粉诱导及其形成的细胞机制研究
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摘要
以铁皮石斛花蕾为材料,研究不同秋水仙素处理对2n花粉诱导的影响,并探讨2n花粉形成的细胞机制。结果表明:用0.1%秋水仙素微量注射长5.53mm、宽2.3mm左右的花蕾,每天注射1次,共注射3次,诱导2n花粉效果最好,2n花粉诱导率达6.22%,2n花粉粒直径比n花粉粒直径增大49%。减数分裂中期Ⅱ发现纺锤体定位异常,表现为平行纺锤体、三级纺锤体,四分体时期观察到三分体和二分体,故纺锤体定位异常可能是铁皮石斛2n配子形成的细胞学机制之一,其2n花粉在遗传上等同于FDR型。
Effects of the various colchicine treatments on 2npollen induction in Dendrobium officinale were carried out in the current work.Cytological mechanism of 2npollen formation were also studied.The results showed that the optimum method for 2npollen induction was injecting 0.1% colchicines into the flower buds at about 5.53 mm long and 2.3mm wide for 3times,once a day,with the highest induction rate up to 6.22%.the diameter of 2npollen was 49%larger than n pollen.The abnormal phenomena like parallel spindles and tripolar spindles at meiotic metaphase Ⅱ,triads and dyads at the tetrad period were observed,so the abnormalities in spindle positioning may be one of the cytological mechanism of 2npollen formation in Den.officinale,and its 2npollens were genetically equivalent to FDR(first division restitution)type.
Effects of the various colchicine treatments on 2npollen induction in Dendrobium officinale were carried out in the current work.Cytological mechanism of 2npollen formation were also studied.The results showed that the optimum method for 2npollen induction was injecting 0.1% colchicines into the flower buds at about 5.53 mm long and 2.3mm wide for 3times,once a day,with the highest induction rate up to 6.22%.the diameter of 2npollen was 49%larger than n pollen.The abnormal phenomena like parallel spindles and tripolar spindles at meiotic metaphase Ⅱ,triads and dyads at the tetrad period were observed,so the abnormalities in spindle positioning may be one of the cytological mechanism of 2npollen formation in Den.officinale,and its 2npollens were genetically equivalent to FDR(first division restitution)type.
引文
[1]国家药典委员会.中华人民共和国药典[M].北京:中国医药科技出版社,2015.
[2]RAMSEY J,SCHCMSKE D W.Pathways,mechanisms,and rates of polyploidy formation in flowering plants[J].Annu.Rev.Ecol.Syst,1998,29:467-501.
[3]DEWITTE A,EECKHAUT T,VAN HUYLENBROECK J,et al.Induction of 2npollen formation in Begoniaby trifluralin and N2O treatments[J].Euphytica,2010,171:283-293.
[4]詹忠根,徐程.秋水仙素诱导铁皮石斛多倍体研究[J].浙江大学学报(理学版),2011,38(3):321-325.ZHAN Z G,XU C.Study on colchiploid of Dendrobium officinale induced by colchicines[J].Journal of Zhejiang University(Science Edition),2011,38(3):321-325.
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[7]YANG J,YAO P Q,LI Y,et al.Induction of 2npollen with colchicine during microsporogenesis in Eucalyptus[J].Euphytica,2016,210:69-78.
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[9]BARBA-GONZALEZ R,LOKKER A C,LIM K B,et al.Use of 2ngametes for the production of sexual polyploids from sterile Oriental×Asiatic hybrids of lilies(Lilium)[J].Theor Appl Genet,2004,109:1 125-1 132.
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[11]CAMADRO E L,SAFFARANO S K,ESPINILLO J C,et al.Cytological mechanisms of 2npollen formation in the wild potato Solanum okadae and pollen-pistil relations with the cultivated potato,Solanum tuberosum with the cultivated potato,Solanum tuberosum[J].Genet Resour Crop Evol,2008,55:471-477.
[12]唐仙英,罗正荣.甜柿2n花粉形成的细胞学机理研究[J].中国农业科学,2002,35(5):585-588.TANG X Y,LUO Z R.Cytology of 2npollen formation in Nonastringent persimmon[J].Scientia Agricultura Sinica,2002,35(5):585-588.
[13]吴红芝,石景峰,郑思乡,等.彩色马蹄莲2n花粉诱导及其三倍体植株的获得[J].农业生物技术学报,2011,19(4):662-668.WU H Z,SHI J F,ZHENG S X,et al.Induction of 2npollen in coloured zantedeschia hybrid and obtaining of its triploid[J].Journal of Agricultural Biotechnology,2011,19(4):662-668.
[14]廖玮玮,邢虎成,揭雨成.植物2n配子诱导及其应用研究进展[J].作物研究,2011,25(6):599-603.LIAO W W,XING H C,JIE Y C.Research progress on plant 2n gamete and its application[J].Crop Research,2011,25(6):599-603.
[15]张正海,康向阳.植物2n配子发生及其遗传标记研究进展[J].遗传,2006,28(1):105-109.ZHANG Z H,KANG X Y.Advances in researches on genetic markers of 2ngametes[J].Hereditas,2006,28(1):105-109.
[16]PELOQUINS S J,BOITEUX L S,CARPUTO D.Meiotic mutants in potato:valuable variants[J].Genentics,1999,153:1 493-1 499.
[17]徐正尧,杨貌仙.黑节草小孢子发生及雄配子体形成的细胞形态学研究[J].云南大学学报,1986,8(3):310-318.XU Z Y,YANG M X.Cyto-morphological studys on the microsporogenesis and the formation of male gametophytes in Dendrobium candidum Wall.ex Lindl[J].Journal of Yunnan University,1986,8(3):310-318.
[2]RAMSEY J,SCHCMSKE D W.Pathways,mechanisms,and rates of polyploidy formation in flowering plants[J].Annu.Rev.Ecol.Syst,1998,29:467-501.
[3]DEWITTE A,EECKHAUT T,VAN HUYLENBROECK J,et al.Induction of 2npollen formation in Begoniaby trifluralin and N2O treatments[J].Euphytica,2010,171:283-293.
[4]詹忠根,徐程.秋水仙素诱导铁皮石斛多倍体研究[J].浙江大学学报(理学版),2011,38(3):321-325.ZHAN Z G,XU C.Study on colchiploid of Dendrobium officinale induced by colchicines[J].Journal of Zhejiang University(Science Edition),2011,38(3):321-325.
[5]VICHIATO M R D,VICHIATO M,PASQUAL M,et al.Tetraploidy induction and identification in Dendrobium nobile L.(Orchidaceae)[J].Revista Ciencia Agronomica,2007,38(4):385-390.
[6]SARATHUM S,HEGELE M,TANTIVIWAT S,et al.Effect of concentration and duration of colchicine treatment on polyploidy induction in Dendrobium scabrilingue L[J].Europ.J.Hort.Sci.,2010,75(3).S:123-127.
[7]YANG J,YAO P Q,LI Y,et al.Induction of 2npollen with colchicine during microsporogenesis in Eucalyptus[J].Euphytica,2016,210:69-78.
[8]范彬.春剑2n花粉的人工诱导[D].成都:四川农业大学,2012.
[9]BARBA-GONZALEZ R,LOKKER A C,LIM K B,et al.Use of 2ngametes for the production of sexual polyploids from sterile Oriental×Asiatic hybrids of lilies(Lilium)[J].Theor Appl Genet,2004,109:1 125-1 132.
[10]XUE Z H,LIU P,LIU M J.Cytological mechanism of 2n pollen formation in Chinese jujube(Ziziphus jujuba Mill.‘Linglingzao’)[J].Euphytica,2011,182(2):231-238.
[11]CAMADRO E L,SAFFARANO S K,ESPINILLO J C,et al.Cytological mechanisms of 2npollen formation in the wild potato Solanum okadae and pollen-pistil relations with the cultivated potato,Solanum tuberosum with the cultivated potato,Solanum tuberosum[J].Genet Resour Crop Evol,2008,55:471-477.
[12]唐仙英,罗正荣.甜柿2n花粉形成的细胞学机理研究[J].中国农业科学,2002,35(5):585-588.TANG X Y,LUO Z R.Cytology of 2npollen formation in Nonastringent persimmon[J].Scientia Agricultura Sinica,2002,35(5):585-588.
[13]吴红芝,石景峰,郑思乡,等.彩色马蹄莲2n花粉诱导及其三倍体植株的获得[J].农业生物技术学报,2011,19(4):662-668.WU H Z,SHI J F,ZHENG S X,et al.Induction of 2npollen in coloured zantedeschia hybrid and obtaining of its triploid[J].Journal of Agricultural Biotechnology,2011,19(4):662-668.
[14]廖玮玮,邢虎成,揭雨成.植物2n配子诱导及其应用研究进展[J].作物研究,2011,25(6):599-603.LIAO W W,XING H C,JIE Y C.Research progress on plant 2n gamete and its application[J].Crop Research,2011,25(6):599-603.
[15]张正海,康向阳.植物2n配子发生及其遗传标记研究进展[J].遗传,2006,28(1):105-109.ZHANG Z H,KANG X Y.Advances in researches on genetic markers of 2ngametes[J].Hereditas,2006,28(1):105-109.
[16]PELOQUINS S J,BOITEUX L S,CARPUTO D.Meiotic mutants in potato:valuable variants[J].Genentics,1999,153:1 493-1 499.
[17]徐正尧,杨貌仙.黑节草小孢子发生及雄配子体形成的细胞形态学研究[J].云南大学学报,1986,8(3):310-318.XU Z Y,YANG M X.Cyto-morphological studys on the microsporogenesis and the formation of male gametophytes in Dendrobium candidum Wall.ex Lindl[J].Journal of Yunnan University,1986,8(3):310-318.