一种沙地苔藓的孢子萌发及愈伤组织诱导
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摘要
本研究以采集自川西北红原县瓦切乡沙化草地的紫色真藓(Bryum purpurascens)成熟孢蒴为材料,研究了其孢子萌发和原丝体发育、不同消毒剂种类、浓度以及不同植物激素处理对其愈伤组织诱导的影响。结果表明:紫色真藓孢蒴最适消毒程序为75%酒精10 s,2%NaClO 20 s (重复2个循环),所获的孢子无菌率为100%,成活率为98.0%。紫色真藓孢子萌发无休眠期,萌发时间为1～2 d,且为多级萌发。以生长20 d原丝体为原材料,采用Knop作为基础培养基,蔗糖浓度为2 g/L,添加0.1 mg/L 6-BA+3 mg/L NAA能够促进愈伤组织的诱导和分化。本研究的结果为紫色真藓快速繁殖体系的建立等提供了科学依据,也为川西北高寒沙化生物结皮等研究提供了参考。
The spore germination, protonema development and the effect of different disinfectants and different concentrations of plant hormone on callus induction were studied, using the mature capsules of Bryum purpurascens collected from the desertification grassland in Waqie, Hongyuan county, northwest Sichuan Province. The results showed that the best disinfectant procedures were 75% alcohol immersion for 10 s and 2% sodium hypochlorite for 20 s with two repeating cycles. The sterile rate of the obtained spores was 100% and the spore survival rate was98.4%. The dormancy stage was absent during spore development of Bryum purpurascens. The germination time was 1～2 d, and the germination was multistage. Using the protonema grown 20 d as the material, the conditions including the basic Knop medium with 2 g/L sucrose, adding 0.1 mg/L 6-BA+3 mg/L NAA could promote the induction and differentiation of callus. The results of the study would provide the scientific basis for the establishment of rapid propagation system of Bryum purpurascens, as well as the important reference for biological soil crust research of the desertification grassland in Northwest Sichuan Province.
The spore germination, protonema development and the effect of different disinfectants and different concentrations of plant hormone on callus induction were studied, using the mature capsules of Bryum purpurascens collected from the desertification grassland in Waqie, Hongyuan county, northwest Sichuan Province. The results showed that the best disinfectant procedures were 75% alcohol immersion for 10 s and 2% sodium hypochlorite for 20 s with two repeating cycles. The sterile rate of the obtained spores was 100% and the spore survival rate was98.4%. The dormancy stage was absent during spore development of Bryum purpurascens. The germination time was 1～2 d, and the germination was multistage. Using the protonema grown 20 d as the material, the conditions including the basic Knop medium with 2 g/L sucrose, adding 0.1 mg/L 6-BA+3 mg/L NAA could promote the induction and differentiation of callus. The results of the study would provide the scientific basis for the establishment of rapid propagation system of Bryum purpurascens, as well as the important reference for biological soil crust research of the desertification grassland in Northwest Sichuan Province.
引文
Belnap J.,and Gillette D.A.,1998,Vulnerability of desert biological soil crusts to wind erosion:the influences of crust development,soil texture,and disturbance,J.Arid Environ.,39(2):133-142
Deng D.Z.,Yan W.X.,Wu B.X.,Zhang W.,Huang X.,and Yu L.F.,2015,The analysis of necessity of consolidate the achievements of grassland des ertification control in Northwest Sichuan,Sichuan Linye Keji(Journal of Sichuan Forestry and Technology),36(1):69-72(邓东周,鄢武先,武碧先,张炜,黄茜,余凌帆,2015,川西北防沙治沙试点示范工程成果巩固必要性分析,四川林业科技,36(1):69-72)
Gao Y.Q.,Ren D.M.,Zhao X.D.,Zhao D.P.,and Sui Y.H.,2016,Tissue culture of Didymodon tectorum,Xibei Zhiwu Xuebao(Acta Botanica Boreali-Occidentalia Sinica),36(9):1900-1904(高叶青,任冬梅,赵小丹,赵东平,隋嫣鸿,2016,短叶对齿藓组织培养的研究,西北植物学报,36(9):1900-1904)
Gonzalez M.L.,Mallon R.,Reinoso J.,and Rodriguez-Oubina J.,2006,In vitro micropropagation and long-term conservation of the endangered moss Splachnum ampullaceum,Biol.Plantarum,50(50):339-345
Li Z.L.,2015,Research on tissue culture and regeneration of Sphagnum,Thesis for M.S.,East China Normal University,Supervisor:Sun Y.,pp.56-57(李泽玲,2015,泥炭藓组织培养及再生现象的研究,硕士学位论文,华东师范大学,导师:孙越,pp.56-57)
Pan Y.T.,Shi D.J.,Yang M.L.,Wu P.C.,and Du G.S.,2005,Callus induction and culture of Physcomitrella patens,Zhiwu Shenglixue Tongxun(Plant Physiology Communication),41(3):293-296(潘一廷,施定基,杨明丽,吴鹏程,杜桂森,2005,小立碗藓愈伤组织诱导和培养,植物生理学通讯,41(3):293-296)
Wang M.,Sha W.,Zhang M.J.,Zhu W.W.,and Xin Y.M.,2015,Effects of high temperature stress on the physiological and biochemical characteristics in Grimmia pilifera,Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),34(6):1290-1295(王曼,沙伟,张梅娟,朱巍巍,信雨萌,2015,高温胁迫对毛尖紫萼藓生理生化特性的影响,基因组学与应用生物学,34(6):1290-1295)
Wei Z.Y.,Sha W.,and Zhang M.J.,2014,Study on tissue culture conditions of Tortula ruralis,Beifang Yuanyi(Northern Horticulture),18(6):114-117(魏志颖,沙伟,张梅娟,2014,山墙藓组织培养条件研究,北方园艺,18(6):114-117)
Wu X.F.,Hu R.Y.,and Li X.D.,2013,Spore germination and callus induction of Atrichum undulatum,Zhiwu Xuebao(Chinese Bulletin of Botany),48(6):651-657(吴小凤,胡若洋,李学东,2013,仙鹤藓的孢子萌发及愈伤组织诱导,植物学报,48(6):651-657)
Xiao B.X.,Chen Q.,Zheng Q.Y.,Yang M.Y.,and Sun H.Y.,2012,The current situation,problem and suggestion on grassland desertification control in Northwest Sichuan,Caoye Yu Xumu(Prataculture&Animal Husbandry),(12):21-23(肖冰雪,陈琴,郑群英,杨满业,孙海燕,2012,川西北草原沙化治理现状、存在的问题及建议,草业与畜牧,(12):21-23)
Xu J.,Bai L.X.,Yang C.,and Zhang P.,2003,Study on diversity and binding-sand effect of moss on biotic crusts of fixed dunes,Zhiwu Shengtai Xuebao(Acta Phytoecologica Sinica),27(4):545-551(徐杰,白良学,杨持,张萍,2003,固定沙丘结皮层藓类植物多样性及固沙作用研究,植物生态学报,27(4):545-551)
Yang Y.S.,Feng W.,Yuan F.,Zhang P.,Ye J.,and Bu C.F.,2015,Key influential factors of rapid cultivation of moss crusts on Loess Plateau,Shuitu Baochi Xuebao(Journal of Soil and Water Conservation),29(4):289-294(杨永胜,冯伟,袁方,张朋,叶菁,卜崇峰,2015,快速培育黄土高原苔藓结皮的关键影响因子,水土保持学报,29(4):289-294)
Zhang N.,Du B.M.,and Ji M.C.,2011,Research progress on the bryophytes tissue culture,Zhejiang Nonglin Daxue Xuebao(Journal of Zhejiang A&F University),28(2):305-313(张楠,杜宝明,季梦成,2011,苔藓植物组织培养研究进展,浙江农林大学学报,28(2):305-313)
Zhang Y.M.,Cao T.,and Pan B.R.,2002,A study on bryophyte associated with formation of soil crust in south fringe of Gurbantunggut Desert in Xinjiang,Xibei Zhiwu Xuebao(Acta Botanica Boreali-Occidentalia Sinica),22(1):18-23(张元明,曹同,潘伯荣,2002,新疆古尔班通古特沙漠南缘土壤结皮中苔藓植物的研究,西北植物学报,22(1):18-23)
Zheng Y.P.,Zhao J.C.,Zhang B.C.,Li L.,and Zhang Y.M.,2009,Advances on ecological studies of algae and mosses in biological soil crust,Zhiwu Xuebao(Chinese Bulletin of Botany),44(3):371-378(郑云普,赵建成,张丙昌,李琳,张元明,2009,荒漠生物结皮中的藻类和苔藓植物研究进展,植物学报,44(3):371-378)
Deng D.Z.,Yan W.X.,Wu B.X.,Zhang W.,Huang X.,and Yu L.F.,2015,The analysis of necessity of consolidate the achievements of grassland des ertification control in Northwest Sichuan,Sichuan Linye Keji(Journal of Sichuan Forestry and Technology),36(1):69-72(邓东周,鄢武先,武碧先,张炜,黄茜,余凌帆,2015,川西北防沙治沙试点示范工程成果巩固必要性分析,四川林业科技,36(1):69-72)
Gao Y.Q.,Ren D.M.,Zhao X.D.,Zhao D.P.,and Sui Y.H.,2016,Tissue culture of Didymodon tectorum,Xibei Zhiwu Xuebao(Acta Botanica Boreali-Occidentalia Sinica),36(9):1900-1904(高叶青,任冬梅,赵小丹,赵东平,隋嫣鸿,2016,短叶对齿藓组织培养的研究,西北植物学报,36(9):1900-1904)
Gonzalez M.L.,Mallon R.,Reinoso J.,and Rodriguez-Oubina J.,2006,In vitro micropropagation and long-term conservation of the endangered moss Splachnum ampullaceum,Biol.Plantarum,50(50):339-345
Li Z.L.,2015,Research on tissue culture and regeneration of Sphagnum,Thesis for M.S.,East China Normal University,Supervisor:Sun Y.,pp.56-57(李泽玲,2015,泥炭藓组织培养及再生现象的研究,硕士学位论文,华东师范大学,导师:孙越,pp.56-57)
Pan Y.T.,Shi D.J.,Yang M.L.,Wu P.C.,and Du G.S.,2005,Callus induction and culture of Physcomitrella patens,Zhiwu Shenglixue Tongxun(Plant Physiology Communication),41(3):293-296(潘一廷,施定基,杨明丽,吴鹏程,杜桂森,2005,小立碗藓愈伤组织诱导和培养,植物生理学通讯,41(3):293-296)
Wang M.,Sha W.,Zhang M.J.,Zhu W.W.,and Xin Y.M.,2015,Effects of high temperature stress on the physiological and biochemical characteristics in Grimmia pilifera,Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),34(6):1290-1295(王曼,沙伟,张梅娟,朱巍巍,信雨萌,2015,高温胁迫对毛尖紫萼藓生理生化特性的影响,基因组学与应用生物学,34(6):1290-1295)
Wei Z.Y.,Sha W.,and Zhang M.J.,2014,Study on tissue culture conditions of Tortula ruralis,Beifang Yuanyi(Northern Horticulture),18(6):114-117(魏志颖,沙伟,张梅娟,2014,山墙藓组织培养条件研究,北方园艺,18(6):114-117)
Wu X.F.,Hu R.Y.,and Li X.D.,2013,Spore germination and callus induction of Atrichum undulatum,Zhiwu Xuebao(Chinese Bulletin of Botany),48(6):651-657(吴小凤,胡若洋,李学东,2013,仙鹤藓的孢子萌发及愈伤组织诱导,植物学报,48(6):651-657)
Xiao B.X.,Chen Q.,Zheng Q.Y.,Yang M.Y.,and Sun H.Y.,2012,The current situation,problem and suggestion on grassland desertification control in Northwest Sichuan,Caoye Yu Xumu(Prataculture&Animal Husbandry),(12):21-23(肖冰雪,陈琴,郑群英,杨满业,孙海燕,2012,川西北草原沙化治理现状、存在的问题及建议,草业与畜牧,(12):21-23)
Xu J.,Bai L.X.,Yang C.,and Zhang P.,2003,Study on diversity and binding-sand effect of moss on biotic crusts of fixed dunes,Zhiwu Shengtai Xuebao(Acta Phytoecologica Sinica),27(4):545-551(徐杰,白良学,杨持,张萍,2003,固定沙丘结皮层藓类植物多样性及固沙作用研究,植物生态学报,27(4):545-551)
Yang Y.S.,Feng W.,Yuan F.,Zhang P.,Ye J.,and Bu C.F.,2015,Key influential factors of rapid cultivation of moss crusts on Loess Plateau,Shuitu Baochi Xuebao(Journal of Soil and Water Conservation),29(4):289-294(杨永胜,冯伟,袁方,张朋,叶菁,卜崇峰,2015,快速培育黄土高原苔藓结皮的关键影响因子,水土保持学报,29(4):289-294)
Zhang N.,Du B.M.,and Ji M.C.,2011,Research progress on the bryophytes tissue culture,Zhejiang Nonglin Daxue Xuebao(Journal of Zhejiang A&F University),28(2):305-313(张楠,杜宝明,季梦成,2011,苔藓植物组织培养研究进展,浙江农林大学学报,28(2):305-313)
Zhang Y.M.,Cao T.,and Pan B.R.,2002,A study on bryophyte associated with formation of soil crust in south fringe of Gurbantunggut Desert in Xinjiang,Xibei Zhiwu Xuebao(Acta Botanica Boreali-Occidentalia Sinica),22(1):18-23(张元明,曹同,潘伯荣,2002,新疆古尔班通古特沙漠南缘土壤结皮中苔藓植物的研究,西北植物学报,22(1):18-23)
Zheng Y.P.,Zhao J.C.,Zhang B.C.,Li L.,and Zhang Y.M.,2009,Advances on ecological studies of algae and mosses in biological soil crust,Zhiwu Xuebao(Chinese Bulletin of Botany),44(3):371-378(郑云普,赵建成,张丙昌,李琳,张元明,2009,荒漠生物结皮中的藻类和苔藓植物研究进展,植物学报,44(3):371-378)