里白孢子体快繁关键技术研究
|
摘要
里白(Diplopterygium glaucum)里白科里白属多年生大型蕨类植物，具有重要的药用价值和观赏价值。本研究以蕨类植物里白的孢子为试验材料，利用组织培养的方法，研究了孢子萌发诱导过程中外植体灭菌、培养基选择、植物生长调节剂的种类、蔗糖浓度、温度、光照、溶液pH值等因素对孢子繁殖的影响，此研究对于蕨类植物的组培快繁及其配子体的发育将对蕨类植物的开发利用、种质保存及种的鉴定、科属的划分、蕨类植物的起源、演化、生态分布等具有重要作用。主要结果如下：
1、消毒灭菌过程中，2%NaClO和70%酒精组合灭菌效果最好，不同的灭菌时间影响到孢子繁殖的污染问题，总体上来看，2%NaClO灭菌30min，而70%酒精灭菌15s，成活率达到85.0%效果最好。
2、不同的培养基类型对里白孢子繁殖的效果不同，几个培养基中，1/2MS培养基的孢子萌发率最高，达到81.1%，萌发时间也最短，在第13d便开始萌发，是较好的培养基。
3、生长素和细胞分裂素对孢子的繁殖具有一定影响。总体上来说看，各种生长素均能促进里白孢子萌发，2,4-D的效果最佳，形成孢子繁殖的时间最短，为7d，并且孢子繁殖效率最高，达到70.0%。各种细胞分裂素中，TDZ的诱导效果最佳，孢子萌发时间也最短，在浓度为2.0mg·L-1诱导率最高，达到61.1%，仅需要8d左右孢子就开始萌发。所有培养基组合中，最佳的培养基组合为改良MS+2,4-D0.2mg·L-1+NAA0.2mg·L-1+TDZ3.0mg·L-1。
4、不同的蔗糖浓度对里白孢子诱导影响不同，孢子诱导率随着蔗糖浓度的不同呈现不同的变化，当蔗糖浓度为30.0g·L-1时，里白孢子诱导率达到最高的91.48%。
5、当光照强度为80μmol·m-2·s-1时，孢子萌发率最高，达到92.17%。孢子需要在光照条件下启动萌发，20-50μmol·m-2·s-1的低光强就能诱导其萌发。
6、温度的高低也会影响到里白孢子的繁殖情况。试验结果表明，25℃时孢子萌发所需时间最短，为7d左右。
7、里白孢子繁殖过程中，培养基溶液pH值5.6时的萌发率最高，为83.2%里白孢子萌发率没有显著性差异，但是当pH值达到7.6时显著降低。
This study using the fern spores of Diplopterygium glaucum as the experimental material,the spore germination induction explant sterilization, selection of culture medium, plant growthregulators, sucrose concentration, temperature, type of light, pH value of solution and other factorson the spore propagation were studied in this text, this text is good to the rapid propagation andutilization of gametophytes, it plays an important role in the genera, divided fern origin, evolution,ecological distribution of Diplopterygium glaucum. The results are showed as follows:
1．The results showed that2%NaClO and70%alcohol combination sterilizing are best.2%NaClO30min70%alcohol sterilization and sterilization,15s, the survival rate of85is best.
2． The results are different when using different medium,1/2MS medium sporegermination rate is highest, reaching81.1%, germination time is the shortest, at the13D began togermination, is a better culture medium.
3. Auxin and cytokinin on spore reproduction has certain effect. On the whole, all sorts ofauxin can promote the white spore germination,2,4-D were the best, formation of spore breedingtime is the shortest,7d, and spore propagation efficiency highest, up to70%. All kinds ofcytokinins, TDZ induction effect is best, spore germination time is the shortest, at a concentrationof2mg/L induce the highest rate, reached61.1%, only need about8D spores begin germination.All medium combinations, the best combination of medium was modified MS+2,4-D0.2mg/L+NAA0.2mg/L+TDZ3mg/L.
4．Different concentrations of sucrose on white spore inducing effects of different inductionrate, spore as the sucrose concentration had different changes, when the sucrose concentration is30.0g/L, white spore inducing rate reaches a maximum of91.48%.
5. When the light intensity of80μmol·m-2·s-1, spore germination rate is the highest,reaching92.17%. Spore is needed in light conditions the initiation ofgermination,20-50/μmol·m-2·s-1low intensity can induce the germination.
6. Temperature also affects the white spore reproduction. The test results show that, at25degrees C spore germination in the shortest time required, is about7d.
7. White spore propagation process, culture medium solution pH value5.6the highestgermination rate,83.2%white spore adorable The rate of no significant difference, but when pH value reaches7.6significantly reduced.
1、消毒灭菌过程中，2%NaClO和70%酒精组合灭菌效果最好，不同的灭菌时间影响到孢子繁殖的污染问题，总体上来看，2%NaClO灭菌30min，而70%酒精灭菌15s，成活率达到85.0%效果最好。
2、不同的培养基类型对里白孢子繁殖的效果不同，几个培养基中，1/2MS培养基的孢子萌发率最高，达到81.1%，萌发时间也最短，在第13d便开始萌发，是较好的培养基。
3、生长素和细胞分裂素对孢子的繁殖具有一定影响。总体上来说看，各种生长素均能促进里白孢子萌发，2,4-D的效果最佳，形成孢子繁殖的时间最短，为7d，并且孢子繁殖效率最高，达到70.0%。各种细胞分裂素中，TDZ的诱导效果最佳，孢子萌发时间也最短，在浓度为2.0mg·L-1诱导率最高，达到61.1%，仅需要8d左右孢子就开始萌发。所有培养基组合中，最佳的培养基组合为改良MS+2,4-D0.2mg·L-1+NAA0.2mg·L-1+TDZ3.0mg·L-1。
4、不同的蔗糖浓度对里白孢子诱导影响不同，孢子诱导率随着蔗糖浓度的不同呈现不同的变化，当蔗糖浓度为30.0g·L-1时，里白孢子诱导率达到最高的91.48%。
5、当光照强度为80μmol·m-2·s-1时，孢子萌发率最高，达到92.17%。孢子需要在光照条件下启动萌发，20-50μmol·m-2·s-1的低光强就能诱导其萌发。
6、温度的高低也会影响到里白孢子的繁殖情况。试验结果表明，25℃时孢子萌发所需时间最短，为7d左右。
7、里白孢子繁殖过程中，培养基溶液pH值5.6时的萌发率最高，为83.2%里白孢子萌发率没有显著性差异，但是当pH值达到7.6时显著降低。
This study using the fern spores of Diplopterygium glaucum as the experimental material,the spore germination induction explant sterilization, selection of culture medium, plant growthregulators, sucrose concentration, temperature, type of light, pH value of solution and other factorson the spore propagation were studied in this text, this text is good to the rapid propagation andutilization of gametophytes, it plays an important role in the genera, divided fern origin, evolution,ecological distribution of Diplopterygium glaucum. The results are showed as follows:
1．The results showed that2%NaClO and70%alcohol combination sterilizing are best.2%NaClO30min70%alcohol sterilization and sterilization,15s, the survival rate of85is best.
2． The results are different when using different medium,1/2MS medium sporegermination rate is highest, reaching81.1%, germination time is the shortest, at the13D began togermination, is a better culture medium.
3. Auxin and cytokinin on spore reproduction has certain effect. On the whole, all sorts ofauxin can promote the white spore germination,2,4-D were the best, formation of spore breedingtime is the shortest,7d, and spore propagation efficiency highest, up to70%. All kinds ofcytokinins, TDZ induction effect is best, spore germination time is the shortest, at a concentrationof2mg/L induce the highest rate, reached61.1%, only need about8D spores begin germination.All medium combinations, the best combination of medium was modified MS+2,4-D0.2mg/L+NAA0.2mg/L+TDZ3mg/L.
4．Different concentrations of sucrose on white spore inducing effects of different inductionrate, spore as the sucrose concentration had different changes, when the sucrose concentration is30.0g/L, white spore inducing rate reaches a maximum of91.48%.
5. When the light intensity of80μmol·m-2·s-1, spore germination rate is the highest,reaching92.17%. Spore is needed in light conditions the initiation ofgermination,20-50/μmol·m-2·s-1low intensity can induce the germination.
6. Temperature also affects the white spore reproduction. The test results show that, at25degrees C spore germination in the shortest time required, is about7d.
7. White spore propagation process, culture medium solution pH value5.6the highestgermination rate,83.2%white spore adorable The rate of no significant difference, but when pH value reaches7.6significantly reduced.
引文
[1]王丽,田京伟,杨建雄等.植物组织培养的研究进展[J].运城高等专科学报,1999,17(6):8-11.
[2]Cutter E G. Experimental induction of buds from fern leaf primordia. Nature,1954,173(4401):440-441.
[3]蒋中海.蕨类植物组织培养研究进展[J].江苏农业科学,2005(5):100-103.
[4]姜述君,冯延江,黎玉梅,等.薇菜孢子人工繁殖的环境条件[J].中国蔬菜,2001,5:11-13.
[5]郝丽珍,邓俊玲.蕨菜孢子萌发及生育进程与诸因素关系研究[J].内蒙古农牧学院学报,1998,19(2):75-80.
[6]曾汉元,丁炳扬.蕨类植物孢子萌发及原叶体发育的观察[J].武汉植物学研究,2004,22(4):368-371.
[7]Wang H M,et al.Spore germination,Prothallus growth and gametophyte development ofAdiantum capillus under invitro and soilless culture conditions[J].Chinahorticulture,1999,(45):353-360.
[8]何义发,易咏梅,赵小兰等.紫萁孢子的无菌培养[J].西南农业大学学报(自然科学版),2004,26(5):613-675.
[9]Fernandez H,Bertrand AM,Ferto I,et al Gametophyte culture in vitro and and theridiogenactivity in Blechum spicant[J].Plant Cell,Tissue and Organ Culture.1997,50(1):71-74.
[10]Higuchi H,et al.Invitro propagation of Nephrolepsis cordifoli Prself[J].ScientiaHortclture,1987）（Higuchi et al，1989；Higuchi et al，1987,32:105-113
[11]翟国燕,珂贾克,朱立新.GA3及不同比例MS培养基对蕨菜孢子萌发的影响[J].中国蔬菜,2007(8):21-23.
[12]程治英,张风雷,兰芹英等.桫椤的快速繁殖与种质保存技术的研究[J].云南植物研究,1991,13(2):181-188.
[13]徐艳,石雷,刘燕等.大叶黑桫椤孢子的无菌培养[J].植物生理学通讯,2004,40(1):72.
[14]程治英，张风雷，兰芹英等.桫椤的快速繁殖与种质保存技术的研究.云南植物研究,1991,13(2):181-188.
[15]Kwa S H,Wee Y C,Loh C S.Production of asporous gametophytes and calli from Pteris vittataL.Pinnae strips cultured invitro.Plant Cell Reports,1991,10:392-393.
[16郭庆勋,沈云霞,宋晓宏,等.猴腿蹄盖蕨孢子萌发和成苗影响因素初探[J].中国农学通报,2007,23(2):343-345.
[17Khoo,SI,Thomas,MB.Studies on the germination of fern spores.Plant Propag.,1980,26:11-15
[18]徐艳,石雷,刘燕等.砷超富集植物—蜈蚣草孢子无菌培养[J].植物生理学通讯,2004,40(6):713.
[19]Dykeman B W,et al.Invitro propagation of the ostrich fern(Matteuacia struthiopteris)[J].Can JPlant Sci.1985,65:1025-1032.
[20]蒋盛军，宋希强，王胜培，刘庆昌.蕨类植物组织培养研究进展,园艺学报,2002,29(增刊):651-656.
[21]徐艳,石雷,刘燕,等.白玉凤尾蕨孢子繁殖技术的研究[J].园艺学报,2005,32(4):658-662.
[22]Douglas D E,et al.A new technique for the culture of fern gametophytes[J].Plant CellReports,1990,(8):632-634.
[23]Cooke R C.Homogenization as an aid in tissue culture propagation of Platycerium andDadallia[J],2001,(7):133-135.
[24]陈星球,叶银根,贾建华,虢华珊.几种陆生薄囊蕨配子体发育过程的初步研究[J].华中师范大学学报(自然科学版),1984,18(1):82-89.
[25]Loescher W H,et al.Development invitro of Nepbrolepis exaltala cv.Bostoniensis runnertissues[J].Physiol Plant,1979,47:250-254.
[26]程治英,张风雷,兰芹英等.桫椤的快速繁殖与种质保存技术的研究[J].云南植物研究,1991,13(2):181-188.
[27]袁艺,田胜尼,叶爱华,等.紫萁快速繁殖技术的研究[J].园艺学报,2002,9(3):247-250.）
[28]Loescher WH,et al.Development in vitro of Neprolepis exaltala cv.Bostoniensis runnertissues[J].Physid Plant,1979,47:250-254
[29]Chatterjee A，porterfield D M，Smith E.Gravity directed calcium in germinating spores ofCeratopteris rhichardii.Planta，2000，205(4):553一560
[30]Guiragossian K.Induction of spore germination in Schizaea pusilia (Schizaeaceae)[J].Amer.J.Bot.,1986,73(11):1588-1594.
[31]Sarah L,Nondorf,Melissa A,et al..The effects of pH,temperature,light intensity,lightquality,and moisture levels on spore germination in cheilanthes feei of SoutheastMissouri[J].American Fern journal,2003,93(2):56-69.
[32]Miller J H，Miller P M. Blue light in the development of the gametophytes and its interaetionwithf ar-red and red light.Amer Jour Bot，1964，51:329一334.
[33]牛俊义,李胜,秦舒浩,等.光照条件及外源物质对蕨菜孢子萌发成苗的影响[J].园艺学报,2002,29(6):584-586.
[34]姜述君,冯延江,黎玉梅,等.薇菜孢子人工繁殖的环境条件[J].中国蔬菜,2001(5):11-13.
[35]Nondorf S L，Dooley M A，Palmieri M，Swatzell L J.The effects of PH，temperature，lightintensity，light quality，and moisture levels on spore germination in Cheilanthes feei of southeastMissouri.Ameriean Fem Jomal,2003,93,56一69.
[36]邵莉楣.观赏蕨类的栽培与用途[M].北京:金盾出版社,1994,30-45.
[37]Savita G,et al.Effects of CTK on spore germination in Adiatum capillus-venevisL.[J].Phytomophyology,1991,141(1):115-120
[38]徐艳,石雷,刘燕,等.白玉凤尾蕨孢子繁殖技术的研究[J].园艺学报,2005,32(4):658-662.
[39]袁艺,田胜尼,叶爱华等.紫萁快速繁殖技术研究[J].园艺学报,2002,9(3):247-250.
[40]罗顺元,王任翔.假鞭叶铁线蕨孢子的组织培养[J].植物生理学通讯,2007,43(1):131-132.）
[41]徐艳,石雷,刘燕等.白玉凤蕨孢子繁殖技术的研究[J].园艺学报,2005,32(4):658-662.）[42]程治英,张风雷,兰芹英等.桫椤的快速繁殖与种质保存技术的研究[J].云南植物研究,1991,13(2):181-188.
[43]鲍敏,吴学明,丁莉.蔗糖和生长辅助物质对蕨孢子人工繁殖的影响[J].青海师范大学学报(自然科学版),2000(3):39-43.
[44]程治英,张风雷,兰芹英等.桫椤的快速繁殖与种质保存技术的研究[J].云南植物研究,1991,13(2):181-188.
[45]蒋胜军,曾霞,王胜培等.海南白桫椤孢子组织培养的研究[J].热带农业科学，2002,22(6):9-12.
[46]吴艳芳,尤敏,王新胜,等.槲蕨配子体发育影响因素的初步研究[J].武汉植物学研究,2005,23(5):461-463.
[47]郭庆勋,沈云霞,宋晓宏,等.猴腿蹄盖蕨孢子萌发和成苗影响因素初探[J].中国农学通报,2007,23(2):343-345.
[48]黄笛,冯玉兰,董丽.银粉背蕨的配子体发育及孢子繁殖技术的研究[J].园艺学报2009,36(9):1345-1352
[49]鲍敏,吴学明,丁莉.蔗糖和生长辅助物质对蕨孢子人工繁殖的影响[J].青海师范大学学报(自然科学版),2000(3):39-43.
[50]Korpelainen H.Labile sex expression in plants[J]． Biol.Rev.Cambridge PhilosophicSociety,1998,73(2):157-180.
[51]牛俊义,李胜,秦舒浩,等.光照条件及外源物质对蕨菜孢子萌发成苗的影响[J].园艺学报,2002,29(6):584-586.
[52]包文美,林孝辉,王全喜,等.东北蕨类植物配子体发育的研究Ⅹ岩蕨科[J].植物研究,1998,18(4):407-413.
[53]刘鲁江,李景富,赵恒田.猴腿蕨孢子囊中孢子不同预处理影响萌发的研究[J].中国林副特产，2007年1期.
[54]徐艳,石雷,刘燕等.大叶黑桫椤孢子的无菌培养[J].植物生理学通讯,2004,40(1):72.
[55]蒋胜军,曾霞,王胜培等.海南白桫椤孢子组织培养的研究[J].热带农业科学，2002,22(6):9-12.
[56]何义发,易咏梅,赵小兰等.紫萁孢子的无菌培养[J].西南农业大学学报(自然科学版),2004,26(5):613-675.
[57]Camloh.M,Ravni kar M,Zel J.Jas monic acid promotes division of ferm protoplasts,eloagation of rhizoids and eady development of gametophytes.Physiologicalplantum,1996,97(4):649-659.
[58]Janss J,Sepple M.In vito anultiplication of Blechnum spp.And pellaeaRotudifolia(forest.)Hook by homoge nization.Scientia Horticalture,1989,38:141-161.
[59]秦延豪,邹宗兰.鸟巢蕨的组织培养[J].植物生理学通讯,2004,40(3):349.
[60]邵莉梅.观赏蕨类的栽培与用途[M].北京:金盾出版社,1994.122-123.
[61]蒋胜军,曾霞,王胜培等.海南白桫椤孢子组织培养的研究[J].热带农业科学，2002,22(6):9-12.
[62]马鑫鑫,杜滨,赵静.荚果蕨叶柄的组织培养及无性系建立[J].安徽农学通报,2007,13(15):21-23.
[63]韦景枫,匡世秀,程友忠,等.贵州六种野生观赏蕨类的组织培养初报[J].贵州林业科技,2007,35(1):55-57.
[64]郭文杰,蔡宣梅,方少忠.华南鳞盖蕨组织培养与假植技术研究[J].中国农学通报,2007,23(1):50-52.
[65]王宏航,李朝森,刘慧琴.观赏蕨类植物组培快繁及其移栽技术[J].江西农业学报,2006,18(5):125-126.
[66]Tshomas.Templeman,Augustus E.Demaggio,Biochemistry of Fern Spore Germination:Globulin Storage Proteins in Matteuccia struthiopteris L.Plant Physiol.1987,85:343-349
[68]鲁雪华,郭文杰,刘润东等.华南鳞盖蕨的组织培养[J].植物生理学通讯,2005,41(4):495.
[69]张开梅,石雷.傅氏凤尾蕨配子体发育的研究[J].植物学通报,2005,22(增刊):50-56.
[70]苏建宇,王俊.蕨的组织培养[J].植物生理学通讯,1996,32(5):16.）
[2]Cutter E G. Experimental induction of buds from fern leaf primordia. Nature,1954,173(4401):440-441.
[3]蒋中海.蕨类植物组织培养研究进展[J].江苏农业科学,2005(5):100-103.
[4]姜述君,冯延江,黎玉梅,等.薇菜孢子人工繁殖的环境条件[J].中国蔬菜,2001,5:11-13.
[5]郝丽珍,邓俊玲.蕨菜孢子萌发及生育进程与诸因素关系研究[J].内蒙古农牧学院学报,1998,19(2):75-80.
[6]曾汉元,丁炳扬.蕨类植物孢子萌发及原叶体发育的观察[J].武汉植物学研究,2004,22(4):368-371.
[7]Wang H M,et al.Spore germination,Prothallus growth and gametophyte development ofAdiantum capillus under invitro and soilless culture conditions[J].Chinahorticulture,1999,(45):353-360.
[8]何义发,易咏梅,赵小兰等.紫萁孢子的无菌培养[J].西南农业大学学报(自然科学版),2004,26(5):613-675.
[9]Fernandez H,Bertrand AM,Ferto I,et al Gametophyte culture in vitro and and theridiogenactivity in Blechum spicant[J].Plant Cell,Tissue and Organ Culture.1997,50(1):71-74.
[10]Higuchi H,et al.Invitro propagation of Nephrolepsis cordifoli Prself[J].ScientiaHortclture,1987）（Higuchi et al，1989；Higuchi et al，1987,32:105-113
[11]翟国燕,珂贾克,朱立新.GA3及不同比例MS培养基对蕨菜孢子萌发的影响[J].中国蔬菜,2007(8):21-23.
[12]程治英,张风雷,兰芹英等.桫椤的快速繁殖与种质保存技术的研究[J].云南植物研究,1991,13(2):181-188.
[13]徐艳,石雷,刘燕等.大叶黑桫椤孢子的无菌培养[J].植物生理学通讯,2004,40(1):72.
[14]程治英，张风雷，兰芹英等.桫椤的快速繁殖与种质保存技术的研究.云南植物研究,1991,13(2):181-188.
[15]Kwa S H,Wee Y C,Loh C S.Production of asporous gametophytes and calli from Pteris vittataL.Pinnae strips cultured invitro.Plant Cell Reports,1991,10:392-393.
[16郭庆勋,沈云霞,宋晓宏,等.猴腿蹄盖蕨孢子萌发和成苗影响因素初探[J].中国农学通报,2007,23(2):343-345.
[17Khoo,SI,Thomas,MB.Studies on the germination of fern spores.Plant Propag.,1980,26:11-15
[18]徐艳,石雷,刘燕等.砷超富集植物—蜈蚣草孢子无菌培养[J].植物生理学通讯,2004,40(6):713.
[19]Dykeman B W,et al.Invitro propagation of the ostrich fern(Matteuacia struthiopteris)[J].Can JPlant Sci.1985,65:1025-1032.
[20]蒋盛军，宋希强，王胜培，刘庆昌.蕨类植物组织培养研究进展,园艺学报,2002,29(增刊):651-656.
[21]徐艳,石雷,刘燕,等.白玉凤尾蕨孢子繁殖技术的研究[J].园艺学报,2005,32(4):658-662.
[22]Douglas D E,et al.A new technique for the culture of fern gametophytes[J].Plant CellReports,1990,(8):632-634.
[23]Cooke R C.Homogenization as an aid in tissue culture propagation of Platycerium andDadallia[J],2001,(7):133-135.
[24]陈星球,叶银根,贾建华,虢华珊.几种陆生薄囊蕨配子体发育过程的初步研究[J].华中师范大学学报(自然科学版),1984,18(1):82-89.
[25]Loescher W H,et al.Development invitro of Nepbrolepis exaltala cv.Bostoniensis runnertissues[J].Physiol Plant,1979,47:250-254.
[26]程治英,张风雷,兰芹英等.桫椤的快速繁殖与种质保存技术的研究[J].云南植物研究,1991,13(2):181-188.
[27]袁艺,田胜尼,叶爱华,等.紫萁快速繁殖技术的研究[J].园艺学报,2002,9(3):247-250.）
[28]Loescher WH,et al.Development in vitro of Neprolepis exaltala cv.Bostoniensis runnertissues[J].Physid Plant,1979,47:250-254
[29]Chatterjee A，porterfield D M，Smith E.Gravity directed calcium in germinating spores ofCeratopteris rhichardii.Planta，2000，205(4):553一560
[30]Guiragossian K.Induction of spore germination in Schizaea pusilia (Schizaeaceae)[J].Amer.J.Bot.,1986,73(11):1588-1594.
[31]Sarah L,Nondorf,Melissa A,et al..The effects of pH,temperature,light intensity,lightquality,and moisture levels on spore germination in cheilanthes feei of SoutheastMissouri[J].American Fern journal,2003,93(2):56-69.
[32]Miller J H，Miller P M. Blue light in the development of the gametophytes and its interaetionwithf ar-red and red light.Amer Jour Bot，1964，51:329一334.
[33]牛俊义,李胜,秦舒浩,等.光照条件及外源物质对蕨菜孢子萌发成苗的影响[J].园艺学报,2002,29(6):584-586.
[34]姜述君,冯延江,黎玉梅,等.薇菜孢子人工繁殖的环境条件[J].中国蔬菜,2001(5):11-13.
[35]Nondorf S L，Dooley M A，Palmieri M，Swatzell L J.The effects of PH，temperature，lightintensity，light quality，and moisture levels on spore germination in Cheilanthes feei of southeastMissouri.Ameriean Fem Jomal,2003,93,56一69.
[36]邵莉楣.观赏蕨类的栽培与用途[M].北京:金盾出版社,1994,30-45.
[37]Savita G,et al.Effects of CTK on spore germination in Adiatum capillus-venevisL.[J].Phytomophyology,1991,141(1):115-120
[38]徐艳,石雷,刘燕,等.白玉凤尾蕨孢子繁殖技术的研究[J].园艺学报,2005,32(4):658-662.
[39]袁艺,田胜尼,叶爱华等.紫萁快速繁殖技术研究[J].园艺学报,2002,9(3):247-250.
[40]罗顺元,王任翔.假鞭叶铁线蕨孢子的组织培养[J].植物生理学通讯,2007,43(1):131-132.）
[41]徐艳,石雷,刘燕等.白玉凤蕨孢子繁殖技术的研究[J].园艺学报,2005,32(4):658-662.）[42]程治英,张风雷,兰芹英等.桫椤的快速繁殖与种质保存技术的研究[J].云南植物研究,1991,13(2):181-188.
[43]鲍敏,吴学明,丁莉.蔗糖和生长辅助物质对蕨孢子人工繁殖的影响[J].青海师范大学学报(自然科学版),2000(3):39-43.
[44]程治英,张风雷,兰芹英等.桫椤的快速繁殖与种质保存技术的研究[J].云南植物研究,1991,13(2):181-188.
[45]蒋胜军,曾霞,王胜培等.海南白桫椤孢子组织培养的研究[J].热带农业科学，2002,22(6):9-12.
[46]吴艳芳,尤敏,王新胜,等.槲蕨配子体发育影响因素的初步研究[J].武汉植物学研究,2005,23(5):461-463.
[47]郭庆勋,沈云霞,宋晓宏,等.猴腿蹄盖蕨孢子萌发和成苗影响因素初探[J].中国农学通报,2007,23(2):343-345.
[48]黄笛,冯玉兰,董丽.银粉背蕨的配子体发育及孢子繁殖技术的研究[J].园艺学报2009,36(9):1345-1352
[49]鲍敏,吴学明,丁莉.蔗糖和生长辅助物质对蕨孢子人工繁殖的影响[J].青海师范大学学报(自然科学版),2000(3):39-43.
[50]Korpelainen H.Labile sex expression in plants[J]． Biol.Rev.Cambridge PhilosophicSociety,1998,73(2):157-180.
[51]牛俊义,李胜,秦舒浩,等.光照条件及外源物质对蕨菜孢子萌发成苗的影响[J].园艺学报,2002,29(6):584-586.
[52]包文美,林孝辉,王全喜,等.东北蕨类植物配子体发育的研究Ⅹ岩蕨科[J].植物研究,1998,18(4):407-413.
[53]刘鲁江,李景富,赵恒田.猴腿蕨孢子囊中孢子不同预处理影响萌发的研究[J].中国林副特产，2007年1期.
[54]徐艳,石雷,刘燕等.大叶黑桫椤孢子的无菌培养[J].植物生理学通讯,2004,40(1):72.
[55]蒋胜军,曾霞,王胜培等.海南白桫椤孢子组织培养的研究[J].热带农业科学，2002,22(6):9-12.
[56]何义发,易咏梅,赵小兰等.紫萁孢子的无菌培养[J].西南农业大学学报(自然科学版),2004,26(5):613-675.
[57]Camloh.M,Ravni kar M,Zel J.Jas monic acid promotes division of ferm protoplasts,eloagation of rhizoids and eady development of gametophytes.Physiologicalplantum,1996,97(4):649-659.
[58]Janss J,Sepple M.In vito anultiplication of Blechnum spp.And pellaeaRotudifolia(forest.)Hook by homoge nization.Scientia Horticalture,1989,38:141-161.
[59]秦延豪,邹宗兰.鸟巢蕨的组织培养[J].植物生理学通讯,2004,40(3):349.
[60]邵莉梅.观赏蕨类的栽培与用途[M].北京:金盾出版社,1994.122-123.
[61]蒋胜军,曾霞,王胜培等.海南白桫椤孢子组织培养的研究[J].热带农业科学，2002,22(6):9-12.
[62]马鑫鑫,杜滨,赵静.荚果蕨叶柄的组织培养及无性系建立[J].安徽农学通报,2007,13(15):21-23.
[63]韦景枫,匡世秀,程友忠,等.贵州六种野生观赏蕨类的组织培养初报[J].贵州林业科技,2007,35(1):55-57.
[64]郭文杰,蔡宣梅,方少忠.华南鳞盖蕨组织培养与假植技术研究[J].中国农学通报,2007,23(1):50-52.
[65]王宏航,李朝森,刘慧琴.观赏蕨类植物组培快繁及其移栽技术[J].江西农业学报,2006,18(5):125-126.
[66]Tshomas.Templeman,Augustus E.Demaggio,Biochemistry of Fern Spore Germination:Globulin Storage Proteins in Matteuccia struthiopteris L.Plant Physiol.1987,85:343-349
[68]鲁雪华,郭文杰,刘润东等.华南鳞盖蕨的组织培养[J].植物生理学通讯,2005,41(4):495.
[69]张开梅,石雷.傅氏凤尾蕨配子体发育的研究[J].植物学通报,2005,22(增刊):50-56.
[70]苏建宇,王俊.蕨的组织培养[J].植物生理学通讯,1996,32(5):16.）