直干桉超级苗组培快繁技术体系研究
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摘要
以直干桉超级苗茎段为外植体,分别采用正交试验、两因素全因子试验和单因素试验逐一探讨直干桉外植体消毒、腋芽诱导、不定芽增殖、生根培养、移栽等组培快繁主要环节中各自的最佳处理,构建直干桉超级苗组织培养的技术体系。结果表明,外植体联合消毒的最佳处理为75%酒精消毒20 s,1%洁尔灭消毒2 min和0.1%升汞消毒5 min,在控制污染率的同时,成活率极显著高于其他处理;暗培养在降低外植体褐化率、提高成活率方面具有极显著效应;腋芽诱导的最佳体系为改良的MS培养基、0.6 mg/L 6-BA和0.5 mg/L NAA,在提高腋芽萌发率的同时,可使芽长、芽壮且结果稳定;不定芽增殖的最佳处理为改良的H培养基、1.0 mg/L 6-BA和0.1 mg/L IBA,不但增大增殖系数,同时还缩短了增殖培养时间;生根培养的最佳处理为改良的White培养基、0.3 mg/L IBA和0.1 mg/L NAA,可极显著提高生根率、缩短生根时间且结果稳定;移栽培育的适宜基质组成为珍珠岩∶腐殖土∶草炭=1∶1∶1,成活率显著高于其他基质组成。
The stem segments of Eucalyptus maideni superior seedlings were used as explants,orthogonal experiment,two-factor full factor experiment and single-factor experiment were adopted,respectively,to explore the best treatment in the main procedures of tissue culture,including disinfection,axillary bud induction,adventitious bud proliferation,rooting culture and transplantation,to establish the technical system for tissue culture of E.maideni superior seedlings.The results showed that the optimal treatments for combined disinfection of explants were 75% alcohol disinfection for 20 s,1% bromo-geramine disinfection for 2 min and 0.1% mercuric chloride disinfection for 5 min.The contamination rate was controlled and the survival rate was significantly higher than those of other treatments.The browning rate was significantly reduced and the survival rate increased markedly through dark culture of explants.The best system for axillary bud induction was the improved MS medium,0.6 mg/L 6-BA and 0.5 mg/L NAA,which could make the buds longer and stronger,and could increase the germination rate of axillary buds.The optimum treatment of adventitious bud proliferation was the improved H medium,1.0 mg/L 6-BA and 0.1 mg/L IBA,which not only remarkably increased the proliferation coefficient but also significantly shortened the breeding time.The optimal treatment of rooting culture was the improved White medium,0.3 mg/L IBA and 0.1 mg/L NAA,which significantly improved rooting rate,shortened rooting time and the test results were very stable.The suitable substrate composition for transplanting cultivation was perlite∶humus soil∶turf=1∶1∶1,the survival rate was significantly higher than that of other substrates.
The stem segments of Eucalyptus maideni superior seedlings were used as explants,orthogonal experiment,two-factor full factor experiment and single-factor experiment were adopted,respectively,to explore the best treatment in the main procedures of tissue culture,including disinfection,axillary bud induction,adventitious bud proliferation,rooting culture and transplantation,to establish the technical system for tissue culture of E.maideni superior seedlings.The results showed that the optimal treatments for combined disinfection of explants were 75% alcohol disinfection for 20 s,1% bromo-geramine disinfection for 2 min and 0.1% mercuric chloride disinfection for 5 min.The contamination rate was controlled and the survival rate was significantly higher than those of other treatments.The browning rate was significantly reduced and the survival rate increased markedly through dark culture of explants.The best system for axillary bud induction was the improved MS medium,0.6 mg/L 6-BA and 0.5 mg/L NAA,which could make the buds longer and stronger,and could increase the germination rate of axillary buds.The optimum treatment of adventitious bud proliferation was the improved H medium,1.0 mg/L 6-BA and 0.1 mg/L IBA,which not only remarkably increased the proliferation coefficient but also significantly shortened the breeding time.The optimal treatment of rooting culture was the improved White medium,0.3 mg/L IBA and 0.1 mg/L NAA,which significantly improved rooting rate,shortened rooting time and the test results were very stable.The suitable substrate composition for transplanting cultivation was perlite∶humus soil∶turf=1∶1∶1,the survival rate was significantly higher than that of other substrates.
引文
[1] 谢耀坚.中国桉树人工林可持续经营战略初探[J].世界林业研究,2003,16(5):58-61.XIE Y J.Primary studies on sustainable management strategy of Eucalyptus plantation in China[J].World Forestry Research,2003,16(5):58-61.(in Chinese)
[2] 陈余美,林彬远.直干桉引种及培育技术研究[J].四川林业科技,2006,27(3):10-14.CHEN Y M,LIN B Y.A study of the intrdocution and cultural techniques of Eucalyetus maidenii[J].Journal of Sichuan Forestry Science and Technology,2006,27(3):10-14.(in Chinese)
[3] 张荣贵,李思广,蒋云东.云南的桉树引种及对其发展状况的剖析[J].西部林业科学,2007,36(3):98-102.ZHANG R G,LI S G,JIANG Y D.Introduction of Eucalyptus in Yunnan and analysis on development status[J].Journal of West China Forestry Science,2007,36(3):98-102.(in Chinese)
[4] 曾德贤,吴子欢,朱仁刚.云南省林业产业发展的良种问题及对策[J].林业调查规划,2010,35(5):57-61.ZENG D X,WU Z H,ZHU R G.Problem of improved seeds and countermeasures of forest industry development inYunnan[J].Forest Inventory and Planning,2010,35(5):57-61.(in Chinese)
[5] 黄海平,郑祥.浅谈弥勒县直干桉栽培现状及发展对策[J].林业建设,2010(3):22-24.
[6] 张永桥,王自全,萧增新.陆良县桉树经营方略初探[J].绿色科技,2014(9):56-57.
[7] 王晓丽,杨再国,曹梦涵,等.蓝桉及直干桉超级苗初步选择研究[J].西南林业大学学报,2018,38(4),89-93.WANG X L,YANG Z G,CAO M H,et al.Preliminary selection of super seedlings of Eucalyptus globules and Eucalyptus maideni[J].Journal of Southwest Forestry University,2018,38(4):89-93.(in Chinese)
[8] 马常耕.世界林木遗传改良研究水平与趋势[J].世界林业研究,1991(1):85-87.
[9] 任辉丽.植物组培技术常见问题及其预防措施[J].南方农业,2015,9(36):20-21.
[10] 王纪忠,蒋婷婷,朱丽丽.植物组培技术存在的问题及解决方法[J].现代农业科技,2012(20):166-167.
[11] 谢耀坚.桉树组织培养研究进展[J].世界林业研究,2000,13(6):14-19.XIE Y J.Advances in tissue culture of Eucalyptus[J].World Forestry Research,2000,13(6):14-19.(in Chinese)
[12] 江海涛.桉树组培快繁研究及其应用进展[J].现代建设,2012,11(7):64-67.
[13] ADAM S.Obtention de racines transformees chez Eucalyptus gunnii H par Agrobacterium rhizogenes[J].Annales de Recherches Silvicoles,Association Foret-Cellulose,1987:7-21.
[14] ARYA H C,SHEKHAWAT N S.Clonal multeplication of tree species in the Thar desert through tissue cultrue[J].Forest Ecol.Manag,1986,16:201-208.
[15] CATHLEEN M,RAJ D,ALEXANDER A M,et al.Development of Eucalyptus tissue culture conditions for improved in vitro plant health and transformability[J].BMC Proceedings 2011,5(7):153.
[16] 沙月娥,吴志华,欧阳乐军,等.粗皮桉的组织培养与植株再生研究[J].南方农业学报,2013,44(9):1511-1516.SHA Y E,WU ZHI H,OUYANG L J,et al.Tissue culture and regeneration of Eucalyptus pellita[J].Journal of Southern Agriculture,2013,44(9):1511-1516.(in Chinese)
[17] 燕丽萍,夏阳,毛秀红,等.邓恩桉的组织培养[J].林业科学,2011,47(5):157-161.YAN L P,XIA Y,MAO X H,et al.Tissue culture of Eucalyptus dunnii[J].Scientia Silvae Sinicae,2011,47(5):157-161.(in Chinese)
[18] 刘均利,刘海鹰,龙汉利,等.柳桉组培快繁技术体系研究[J].四川林业科技,2016,37(4):74-78.LIU J L,LIU H Y,LONG H L,et al.A study of tissue culture of Eucalyptus saligna[J].Journal of Sichuan Forestry Science and Technology,2016,37(4):74-78.(in Chinese)
[19] 刘均利,郭洪英,杨晓蓉,等.柠檬桉的组培快繁技术研究[J].桉树科技,2013,30(4):19-24.LIU J L,GUO H Y,YANG X R,et al.Studies on tissue culture of Eucalyptus citriodora[J].Eucalypt Science & Technology,2013,30(4):19-24.(in Chinese)
[20] 刘云彩,陈芳,吴丽圆,等.直干桉组织培养[J].云南林业科技,1996(3):12-18.LIU Y C,CHEN F,WU L Y,et al.Study on tissue culture of Eucalyptus maidenii[J].Journal of Yunnan Forestry Science and Technology,1996(3):12-18.(in Chinese)
[21] 佘小涵.直杆蓝桉组织培养繁殖育苗试验研究[J].福建林业科技,2002,29(3):26-29.SHE X H.Experimental studies on the seedling raising of tissue culture propagation of Eucalyptus labill[J].Journal of Fujian Forestry Science and Technology,2002,29(3):26-29.(in Chinese)
[22] 谢志亮,吴振旺.木本植物组培褐化研究进展[J].中国南方果树,2013,42(5):42-46.
[23] 张宏平,姬爱国,和林涛.植物组培快繁褐化现象研究进展[J].农业工程,2013,3(5):128-132.ZHANG H P,JI A G,HE L T.Research progress of browning phenomenon about plant tissue culture[J].Agricultural Engineering,2013,3(5):128-132.(in Chinese)
[24] 雷亚灵,李周岐.八仙花茎段组织培养技术研究[J].西北林学院学报,2008,23(4):101-103.LEI Y L.A study on tissue culture for Hydrangea Macropphylla[J].Journal of Northwest Forestry University,2008,23(4):101-103.(in Chinese)
[25] 唐豆豆,李厚华,张延龙,等.‘凤丹’牡丹组织培养研究[J].西北林学院学报,2016,31(2):160-166.TANG D D,LI H H,ZHANG Y L et al.Studies on tissue culture of Paeonia ostii‘Feng Dan’[J].Journal of Northwest Forestry University,2016,31(2):160-166.(in Chinese)
[26] 覃林海,韦素婕,王芳,等.不同培养基对红心杉组培苗增殖及其生理的影响[J].西北林学院学报,2017,32(3):122-127.QIN L H,WEI S J,WANG F.Effects of different media on the seedlings growth and physiology in Cunninghamia Lanceolata[J].Journal of Northwest Forestry University,2017,32(3):122-127.(in Chinese)
[2] 陈余美,林彬远.直干桉引种及培育技术研究[J].四川林业科技,2006,27(3):10-14.CHEN Y M,LIN B Y.A study of the intrdocution and cultural techniques of Eucalyetus maidenii[J].Journal of Sichuan Forestry Science and Technology,2006,27(3):10-14.(in Chinese)
[3] 张荣贵,李思广,蒋云东.云南的桉树引种及对其发展状况的剖析[J].西部林业科学,2007,36(3):98-102.ZHANG R G,LI S G,JIANG Y D.Introduction of Eucalyptus in Yunnan and analysis on development status[J].Journal of West China Forestry Science,2007,36(3):98-102.(in Chinese)
[4] 曾德贤,吴子欢,朱仁刚.云南省林业产业发展的良种问题及对策[J].林业调查规划,2010,35(5):57-61.ZENG D X,WU Z H,ZHU R G.Problem of improved seeds and countermeasures of forest industry development inYunnan[J].Forest Inventory and Planning,2010,35(5):57-61.(in Chinese)
[5] 黄海平,郑祥.浅谈弥勒县直干桉栽培现状及发展对策[J].林业建设,2010(3):22-24.
[6] 张永桥,王自全,萧增新.陆良县桉树经营方略初探[J].绿色科技,2014(9):56-57.
[7] 王晓丽,杨再国,曹梦涵,等.蓝桉及直干桉超级苗初步选择研究[J].西南林业大学学报,2018,38(4),89-93.WANG X L,YANG Z G,CAO M H,et al.Preliminary selection of super seedlings of Eucalyptus globules and Eucalyptus maideni[J].Journal of Southwest Forestry University,2018,38(4):89-93.(in Chinese)
[8] 马常耕.世界林木遗传改良研究水平与趋势[J].世界林业研究,1991(1):85-87.
[9] 任辉丽.植物组培技术常见问题及其预防措施[J].南方农业,2015,9(36):20-21.
[10] 王纪忠,蒋婷婷,朱丽丽.植物组培技术存在的问题及解决方法[J].现代农业科技,2012(20):166-167.
[11] 谢耀坚.桉树组织培养研究进展[J].世界林业研究,2000,13(6):14-19.XIE Y J.Advances in tissue culture of Eucalyptus[J].World Forestry Research,2000,13(6):14-19.(in Chinese)
[12] 江海涛.桉树组培快繁研究及其应用进展[J].现代建设,2012,11(7):64-67.
[13] ADAM S.Obtention de racines transformees chez Eucalyptus gunnii H par Agrobacterium rhizogenes[J].Annales de Recherches Silvicoles,Association Foret-Cellulose,1987:7-21.
[14] ARYA H C,SHEKHAWAT N S.Clonal multeplication of tree species in the Thar desert through tissue cultrue[J].Forest Ecol.Manag,1986,16:201-208.
[15] CATHLEEN M,RAJ D,ALEXANDER A M,et al.Development of Eucalyptus tissue culture conditions for improved in vitro plant health and transformability[J].BMC Proceedings 2011,5(7):153.
[16] 沙月娥,吴志华,欧阳乐军,等.粗皮桉的组织培养与植株再生研究[J].南方农业学报,2013,44(9):1511-1516.SHA Y E,WU ZHI H,OUYANG L J,et al.Tissue culture and regeneration of Eucalyptus pellita[J].Journal of Southern Agriculture,2013,44(9):1511-1516.(in Chinese)
[17] 燕丽萍,夏阳,毛秀红,等.邓恩桉的组织培养[J].林业科学,2011,47(5):157-161.YAN L P,XIA Y,MAO X H,et al.Tissue culture of Eucalyptus dunnii[J].Scientia Silvae Sinicae,2011,47(5):157-161.(in Chinese)
[18] 刘均利,刘海鹰,龙汉利,等.柳桉组培快繁技术体系研究[J].四川林业科技,2016,37(4):74-78.LIU J L,LIU H Y,LONG H L,et al.A study of tissue culture of Eucalyptus saligna[J].Journal of Sichuan Forestry Science and Technology,2016,37(4):74-78.(in Chinese)
[19] 刘均利,郭洪英,杨晓蓉,等.柠檬桉的组培快繁技术研究[J].桉树科技,2013,30(4):19-24.LIU J L,GUO H Y,YANG X R,et al.Studies on tissue culture of Eucalyptus citriodora[J].Eucalypt Science & Technology,2013,30(4):19-24.(in Chinese)
[20] 刘云彩,陈芳,吴丽圆,等.直干桉组织培养[J].云南林业科技,1996(3):12-18.LIU Y C,CHEN F,WU L Y,et al.Study on tissue culture of Eucalyptus maidenii[J].Journal of Yunnan Forestry Science and Technology,1996(3):12-18.(in Chinese)
[21] 佘小涵.直杆蓝桉组织培养繁殖育苗试验研究[J].福建林业科技,2002,29(3):26-29.SHE X H.Experimental studies on the seedling raising of tissue culture propagation of Eucalyptus labill[J].Journal of Fujian Forestry Science and Technology,2002,29(3):26-29.(in Chinese)
[22] 谢志亮,吴振旺.木本植物组培褐化研究进展[J].中国南方果树,2013,42(5):42-46.
[23] 张宏平,姬爱国,和林涛.植物组培快繁褐化现象研究进展[J].农业工程,2013,3(5):128-132.ZHANG H P,JI A G,HE L T.Research progress of browning phenomenon about plant tissue culture[J].Agricultural Engineering,2013,3(5):128-132.(in Chinese)
[24] 雷亚灵,李周岐.八仙花茎段组织培养技术研究[J].西北林学院学报,2008,23(4):101-103.LEI Y L.A study on tissue culture for Hydrangea Macropphylla[J].Journal of Northwest Forestry University,2008,23(4):101-103.(in Chinese)
[25] 唐豆豆,李厚华,张延龙,等.‘凤丹’牡丹组织培养研究[J].西北林学院学报,2016,31(2):160-166.TANG D D,LI H H,ZHANG Y L et al.Studies on tissue culture of Paeonia ostii‘Feng Dan’[J].Journal of Northwest Forestry University,2016,31(2):160-166.(in Chinese)
[26] 覃林海,韦素婕,王芳,等.不同培养基对红心杉组培苗增殖及其生理的影响[J].西北林学院学报,2017,32(3):122-127.QIN L H,WEI S J,WANG F.Effects of different media on the seedlings growth and physiology in Cunninghamia Lanceolata[J].Journal of Northwest Forestry University,2017,32(3):122-127.(in Chinese)
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