多花相思离体培养与再生体系的建立
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摘要
多花相思(Acacia floribunda (Vent.) Willd.)自然分布于澳大利亚昆士兰、新南威士和维多利亚等州,适应性较强,有较高的观赏价值。本论文以自然状态下多花相思的种子为基本材料,从无菌体系的建立、继代培养、生根诱导、炼苗移栽等各个环节展开多花相思无菌体系的建立及快繁技术的研究;同时以多花相思种子苗下胚轴以上部分为材料,进行愈伤组织的诱导,并从愈伤组织分化出苗。主要研究结果如下:
1.多花相思无菌体系的建立。采用1.84g/mL浓硫酸处理种子10min效果最好,发芽率可达90%。然后用75%酒精浸泡10s,0.1%升汞浸泡15min,无菌水冲洗7遍,可以使污染率控制在3%,最后接种到MS培养基中,生长状况良好。
2.采用正交试验设计,考虑不同植物生长调节剂对多花相思小苗增殖的影响,筛选出最佳一代培养基为MS+KT0.4mg/L+IBA0.5mg/L +BA0.5mg/L +蔗糖30g/L,30d后增殖数可以达到2.914;最佳二次继代培养基为MS培养基+KT0.3mg/L+BA1.0mg/L+IBA1.5mg/L +AC1.2g/L +蔗糖30g/L;30d后增殖数可以达到4.179;最佳三次及多次继代培养基为MS+BA0.2mg/L+IBA0.3mg/L +蔗糖20g/L。30d后增殖数可以达到3.309。
3.多花相思小苗生根诱导。采用正交试验设计,考虑不同种类和浓度生长素对多花相思小苗生根的影响,筛选出最佳生根培养基为MS+IBA0.3mg/L +IAA0.1mg/L +蔗糖30g/L。
4.多花相思组培苗的炼苗和移栽。实验结果表明:黄心土:泥炭土=2:1的效果最好,成活率达到85.5%。
5.多花相思愈伤组织体系的建立。以多花相思种子苗下胚轴以上部分为愈伤组织诱导材料,筛选出愈伤组织的最佳诱导培养基为MS+ BA1.0mg/L+KT0.6mg/L +IBA0.6mg/L+蔗糖30g/L。愈伤组织的最佳继代培养基为MS+ BA1.0mg/L+KT1.0mg/L +IBA0.6mg/L+蔗糖30g/L。当光照强度为1200Lx时,愈伤组织长势最好。
6 .多花相思愈伤组织培养再生植株。以多次继代的愈伤组织为基本材料,在MS+ TDZ0.01mg/L+NAA0.2mg/L +蔗糖30g/L培养基中,可使愈伤组织分化率达到89%,将分化愈伤组织转接在MS+ TDZ0.01mg/L+NAA0.2mg/L +蔗糖30g/L培养基上,再生率为98%。在1/2MS +IBA0.5mg/L+NAA0.1mg/L+AC1.2g/L+20g/L蔗糖培养基上,30天后,小苗增殖数为1.41。
Acacia floribunda (Vent.) Willd. grows in Queensland Australia, New South , WalesVictoria ect. It has good adaptation, high appreciation value. This experiment chose Acacia floribunda (Vent.) Willd.’s seeds which grew in natural state as basic material, researching about every steps of seeds axenic bourgeon, proliferation, root induction, transplantion etc. At the same time, The parts which above the seed’s hypocotyl were used as the initial material for the inducement of calli, calli systems were established, and polarized seedling from calli. The main results were described as follows:
1. The establishment of Acacia floribunda (Vent.) Willd . axenic system.Seed capsule was rigid and compact, deep in oil of vitriol for 10 minutes, reached 90% burgeon rate. The result showed that the method which 75% alcohol disinfected 10 seconds first, then deeped in 0.1%HgCl2 for 15 minutes, and axenic water washed 7 times last was best. It can control the pollution rate below 3%. Next, the seeds were inocu ated to the medium of MS , kept vigorous growth of seeds.
2. The proliferation of Acacia floribunda (Vent.) Willd. axenic seedlings. Considered the influence of different plant growth plant growth regulators on the proliferation, which orthogonal design, screened out the medium was MS supplemented with KT0.4mg/L, IBA0.5mg/L, BA0.5mg/L, sugar 30g/L, and the proliferation rate could reach 2.914 by 30d later. In the second repetitious cultivation, the medium was MS supplemented KT0.3mg/L,BA1.0mg/L ,IBA1.0mg/L, AC1.2g/L ,sugar 30g/L and the proliferation rate could reach 4.179 by 30d later. In the third and more times proliferation, the medium was MS+BA0.2mg/L +IBA0.3mg/L +sugar20mg/L and the proliferation rate could reach3.309 by 30d later.
3. Rootage of Acacia floribunda (Vent.) Willd. Thinked about the influence of different kind and concontraction plant growth regulators on the axenic seedlings root rate, screened the medium was MS supplemented with IBA0.3mg/L, IAA0.1mg/L and sugar 30g/L.
4. The transplantion of Acacia floribunda (Vent.) Willd. Analysis the results of the survival rate of the different Transplanted stroma of Acacia. f loribunda (Vent.) Willd., it showed that ocher:turf=2:1was best and the survival rate can reach 85.5%.
5. The establishment of Acacia floribunda (Vent.) Willd. Calli system. Chose the parts which above seed’s hypocotyl as basic material for the inducement of calli, under the light, consider the influence by different kind and concentration of plant growth regulators and accretion, the medium was MS supplemented with BA1.0mg/L, KT 0.6mg/L, IBA0.6mg/L and sugar 30g/L,and the proliferation medium was MS supplemented with BA1.0mg/L,KT1.0mg/L, IBA0.6mg/L and sugar 30g/L .When the light was 1200lx, the calli grew well.
6. The generation of calli. Under the light of 1200lx, considering the influence of plant growthplant growth regulators and accretion. MS medium supplemented with TDZ0.01mg/L,NAA0.2mg/L, and sugar 30g/L, the calli provenance could reach 89%.MS supplemented with TDZ 0.01 mg/L, NAA0.2mg/L, and sugar 30g/L. the regeneration rate of calli can reach 98%. MS supplemented withTDZ0.01mg/L,NAA0.2mg/L, and sugar 30g/L. the regeneration rate of calli can reach 98%. 1/2MS supplemented with IBA 0.5 mg/ L ,NAA0.1mg/L, AC1.2g/L and sugar 20g/L. Recorded 30d later, the proliferation rate could reach 1.41 .
1.多花相思无菌体系的建立。采用1.84g/mL浓硫酸处理种子10min效果最好,发芽率可达90%。然后用75%酒精浸泡10s,0.1%升汞浸泡15min,无菌水冲洗7遍,可以使污染率控制在3%,最后接种到MS培养基中,生长状况良好。
2.采用正交试验设计,考虑不同植物生长调节剂对多花相思小苗增殖的影响,筛选出最佳一代培养基为MS+KT0.4mg/L+IBA0.5mg/L +BA0.5mg/L +蔗糖30g/L,30d后增殖数可以达到2.914;最佳二次继代培养基为MS培养基+KT0.3mg/L+BA1.0mg/L+IBA1.5mg/L +AC1.2g/L +蔗糖30g/L;30d后增殖数可以达到4.179;最佳三次及多次继代培养基为MS+BA0.2mg/L+IBA0.3mg/L +蔗糖20g/L。30d后增殖数可以达到3.309。
3.多花相思小苗生根诱导。采用正交试验设计,考虑不同种类和浓度生长素对多花相思小苗生根的影响,筛选出最佳生根培养基为MS+IBA0.3mg/L +IAA0.1mg/L +蔗糖30g/L。
4.多花相思组培苗的炼苗和移栽。实验结果表明:黄心土:泥炭土=2:1的效果最好,成活率达到85.5%。
5.多花相思愈伤组织体系的建立。以多花相思种子苗下胚轴以上部分为愈伤组织诱导材料,筛选出愈伤组织的最佳诱导培养基为MS+ BA1.0mg/L+KT0.6mg/L +IBA0.6mg/L+蔗糖30g/L。愈伤组织的最佳继代培养基为MS+ BA1.0mg/L+KT1.0mg/L +IBA0.6mg/L+蔗糖30g/L。当光照强度为1200Lx时,愈伤组织长势最好。
6 .多花相思愈伤组织培养再生植株。以多次继代的愈伤组织为基本材料,在MS+ TDZ0.01mg/L+NAA0.2mg/L +蔗糖30g/L培养基中,可使愈伤组织分化率达到89%,将分化愈伤组织转接在MS+ TDZ0.01mg/L+NAA0.2mg/L +蔗糖30g/L培养基上,再生率为98%。在1/2MS +IBA0.5mg/L+NAA0.1mg/L+AC1.2g/L+20g/L蔗糖培养基上,30天后,小苗增殖数为1.41。
Acacia floribunda (Vent.) Willd. grows in Queensland Australia, New South , WalesVictoria ect. It has good adaptation, high appreciation value. This experiment chose Acacia floribunda (Vent.) Willd.’s seeds which grew in natural state as basic material, researching about every steps of seeds axenic bourgeon, proliferation, root induction, transplantion etc. At the same time, The parts which above the seed’s hypocotyl were used as the initial material for the inducement of calli, calli systems were established, and polarized seedling from calli. The main results were described as follows:
1. The establishment of Acacia floribunda (Vent.) Willd . axenic system.Seed capsule was rigid and compact, deep in oil of vitriol for 10 minutes, reached 90% burgeon rate. The result showed that the method which 75% alcohol disinfected 10 seconds first, then deeped in 0.1%HgCl2 for 15 minutes, and axenic water washed 7 times last was best. It can control the pollution rate below 3%. Next, the seeds were inocu ated to the medium of MS , kept vigorous growth of seeds.
2. The proliferation of Acacia floribunda (Vent.) Willd. axenic seedlings. Considered the influence of different plant growth plant growth regulators on the proliferation, which orthogonal design, screened out the medium was MS supplemented with KT0.4mg/L, IBA0.5mg/L, BA0.5mg/L, sugar 30g/L, and the proliferation rate could reach 2.914 by 30d later. In the second repetitious cultivation, the medium was MS supplemented KT0.3mg/L,BA1.0mg/L ,IBA1.0mg/L, AC1.2g/L ,sugar 30g/L and the proliferation rate could reach 4.179 by 30d later. In the third and more times proliferation, the medium was MS+BA0.2mg/L +IBA0.3mg/L +sugar20mg/L and the proliferation rate could reach3.309 by 30d later.
3. Rootage of Acacia floribunda (Vent.) Willd. Thinked about the influence of different kind and concontraction plant growth regulators on the axenic seedlings root rate, screened the medium was MS supplemented with IBA0.3mg/L, IAA0.1mg/L and sugar 30g/L.
4. The transplantion of Acacia floribunda (Vent.) Willd. Analysis the results of the survival rate of the different Transplanted stroma of Acacia. f loribunda (Vent.) Willd., it showed that ocher:turf=2:1was best and the survival rate can reach 85.5%.
5. The establishment of Acacia floribunda (Vent.) Willd. Calli system. Chose the parts which above seed’s hypocotyl as basic material for the inducement of calli, under the light, consider the influence by different kind and concentration of plant growth regulators and accretion, the medium was MS supplemented with BA1.0mg/L, KT 0.6mg/L, IBA0.6mg/L and sugar 30g/L,and the proliferation medium was MS supplemented with BA1.0mg/L,KT1.0mg/L, IBA0.6mg/L and sugar 30g/L .When the light was 1200lx, the calli grew well.
6. The generation of calli. Under the light of 1200lx, considering the influence of plant growthplant growth regulators and accretion. MS medium supplemented with TDZ0.01mg/L,NAA0.2mg/L, and sugar 30g/L, the calli provenance could reach 89%.MS supplemented with TDZ 0.01 mg/L, NAA0.2mg/L, and sugar 30g/L. the regeneration rate of calli can reach 98%. MS supplemented withTDZ0.01mg/L,NAA0.2mg/L, and sugar 30g/L. the regeneration rate of calli can reach 98%. 1/2MS supplemented with IBA 0.5 mg/ L ,NAA0.1mg/L, AC1.2g/L and sugar 20g/L. Recorded 30d later, the proliferation rate could reach 1.41 .
引文
[1]叶永昌,朱剑云,李果惠,等.3种观赏相思的引种栽培试验[J].广东林业科技,2001,17(4):16-20.
[2]王忠.植物生理学[M].北京:中国农业出版,2003:323.
[3] Niu G,Kozai T,Kitaya Y.Simulation of the time course of CO2 concentration inthe culture vessel and net pHotosynthetic rate of Cymbidium planlets[J].Transactions of the ASAE,1996,39(4):1567-1573.
[4]Kozai T,Fujiwara K,Kitaya Y.Modeling measurement and control in plant tissue culture [J].Acta Hort,1993,343:68-78.
[5]Afreen Supporting material affects the growth and development of in vitrosweet potato plantlets cultured pHotoautop Hical1y [J].In Vitro Cell Biol Plant,l999,35:470-476.
[6]王蒂,李友勇.应用生物技术[M].中国农业出版社,1997,23-45.
[7]吴丽君.木木植物组织培养技术在林业科研与生产中的应用与局限[J],福建林业科技,2003, 30(1):67-69.
[8]施寄森.迎接21世纪现代林木生物技术育种的挑战[J].南京林业大学学报,2000,22(1):1-6.
[9]韩一凡.德国林木生物技术研究现状[J].世界林业研究,1993,2: 23-26.
[10]杜克久,郑均宝,徐振华,等.741杨器官、体细胞胚胎发生的细胞组织学及生理学的研究[J].林业科学,1998,34(6):99-104.
[11]FAN Guo-giang,ZHAI Xiao-giao,ZHAI Cui-uan,BI Hui-tao,Callus induction from leaves of different paulownia species and its plantlet regeneration [J].Journal of Forestry Research,2001,12(4):209-214.
[12]范国强,翟晓巧,毕会涛,等.金丝橄的离体培养和植株再生[J].植物生理学通讯,2002,38(4):359.
[13]JAIN S M,GUPTA P K,NEWTON R J.Somatic embryogenesis in woody plants.Vol 1-3,Kluwer Academic Publishers, Dordrecht,The Netherlands.1995:335-341.
[14]赵沛基,彭丽萍,甘烦远,等.刺果番荔枝的组织培养[J].植物生理学通讯,2000,36(2):137-138.
[15]唐巍,欧阳藩,郭仲深.针叶树体细胞无性系研究和应用进展[J].生物工程进展,1997,17(4):2-9.
[16]王仲英,王艺,童德中.生物技术在果树上的应用[J].世界农业,1997,12(24):20-23.
[17]孙清荣等.巴城桃子叶不定植株再生[J].山东农业大学学报,1997,30(4):395-397.
[18]Roche T D,Long R D,and Sayegh A J. Commercial-scale pHotoautotropHic micropropagations in Irish agriculture, horticulture and forestry[J].Acta Hort,1996,440:515-520.
[19]Avila A DE L,Pereyra S M,Arguello J A.Nitrogen Concentration and Proportion of NH4+-N Afect Potato Cultivar Response Media[J].HortScience,1998,33(2):336-338.
[20]李世承,李晶,伶凤琴,等.山杨、大青杨、毛白杨组织培养的研究[J].辽宁大学学报(自然科学版),1995,22(2):59-62.
[21]Attree SM,FowkeL C.Embryogeny of gymnosperms:advances in synthetic see technology of conifers[J].Plant Cell Tiss.Org.Cult., 1993,35:1-35.
[22]王傲雪,李景富.植物体细胞胚状体的诱导研究及应用[J].黑龙江农业科学,1999,2:40-41.
[23]王水,贾勇炯,魏峰,等.云南豆杉的组织培养及植株再生[J].云南植物研究,1997,19(4):407-410.
[24]Michio K,Masakatu O,Michiko A,et a1.The effect of carbon dioxide enrichment,and light in tensity on growth, photosynthesis,and transpiration of carliflower plantlets cultured in vitro pHotoautotropHically and photomixotropHically[J].J Amer Soc HortSci,1998,123(2):176-181.
[25]王秀松,胡东坡等.杜仲愈伤组织的诱导及植株再生的研究[J].西北林学院学报,1994,9(4):32-35.
[26]郑晴霞,王凤翱.绒毛皂英离体茎段培养及再生植株的细胞组织学研究[J].湖南农学院学报,1994,20(3):244-248.
[27]李玲,韩一凡.杨树耐盐突变体筛选的研究[J].林业科学,1990,26(4):359-362.
[28]Y.Desjardins.PHotosynthesis in Vitro on the Factors Regulating CO2 Assimilation in Micropropagation Systems[J].Acta Hortculture,1995(393):45-61.
[29]吴清,闰勇,梁国鲁.杨桃胚乳愈伤组织的诱导和三倍体植株再生[J].热带作物学报,2002,23(2):54-57.
[30]何业华,胡芳名,谢碧霞.经济林木离体培养研究进展[J].中南林学院学报,2000,20(1):31-39.
[31]杜雪梅,李登科.落叶果树生物技术育种研究进展[J].果树学报,2002,(5):286-291.
[32]Seko Y, Nishimura M. Effects of CO2 and light on survival and growth of rice regenerates in vitro on sugar-free medium [J]. Plant cell,tissue and organ culture,1997,46 (3): 257-264.
[33]容世清.桉树无性系湛江6号的工厂化微繁研究初报[J].桉树科技,1995 (2):58-59.
[34]王丽娟,田京伟,杨建雄,等.植物组织培养的研究进展[J].运城高等师范专科学校学报,1999,17(3):8-11.
[35]黄维南.固氮树种组织和细胞培养的种类名录[J].亚热带植物通讯,1998. 27(1):21-25.
[36]施寄森.迎接21世纪现代林木生物技术育种的挑战[J].南京林业大学学报, 2000. 24 (1):1-6.
[37]梁一池,杨华.植物组织培养技术的研究进展[J].福建林学院学报,2002, 22 (1): 93-96.
[38]贾利敏,张红梅,贾利欣.玉米组织培养和植株再生的研究现状[J].内蒙古农业科技, 2002,12(3):10-13.
[39]祁永斌,李和平.不同小麦品种愈伤组织诱导和再生体系建立[J].武汉植物学研究,2005,23(3):227-232.
[40]李雪,韩烈保.多年生黑麦草愈伤组织诱导与芽分化的研究[J].草业科学,2005,22(5):45-47.
[41]吴桂胜,胡鸢雷等.剪股颖愈伤组织诱导与植株再生[J].生物技术通报,2006,(4):101-102.
[42]谢启鑫.君迁子叶片培养再生植株的研究[J].中国农业科学, 2008,41(2):607-612.
[43]苗永美.菘蓝两种外植体愈伤组织诱导[J].生物科技通报, 2008, 4(2):45-48.
[44]苏明申,林顺权.荔枝愈伤组织诱导及其体细胞胚胎发生研究初报[J].华南热带农业大学学报,2004,15(9):26-28.
[45]范国强,黎明.悬铃木体细胞胚胎发生及植株再生[J].林业科学,2004,40(3):28-29.
[46]于志水,张志毅.杨树体细胞胚胎发生研究概况[J].东北林业大学学报,2005,33(1):21-23.
[47]姬飞腾,李凤兰.香果树体细胞胚胎发生[J].植物生理学通讯,2005,41(5):19-21.
[48]Attree.S.M,Fowke.L.C. Embryogency of gymnosperms.Plant Cell Tiss Organ Cult,1993,35:1-35.
[49]Jain.S. M, Gupta .P. K,Newton .R. J.Somatic embryogenesis in woody plants. Vol 1-3, Kluwer Academic Publishers, Dordrecht,The Netherlands.1995:335-341.
[50]Tautorus .J .E, Fowke .L .C,Dunstand .I.Somatic embryogenesis in conifers.Candian Joural of Bocany,1991,69:1873-1899
[51]张存旭.栓皮栎体细胞发生的细胞组织学观察[J].植物生理和分子生物学学报, 2007, 33(1) :33-38.
[52]萧洪东.红花银桦组织培养与快速繁殖[J].植物生理学通报, 2008,44(01):34-41.
[53]张宏伟,黄学林,傅家瑞,等.大叶相思、马占相思腋芽培养和植株再生[J].热带亚热带植物学报,1995,3(3):62-68.
[54]蔡玲,王以红,吴幼媚,等.马尖相思离体培养再生植株的研究[J].广西林业科学,1999,03: 258-261.
[55]高洁,玉玲.大叶相思茎段腋芽组织培养技术初探[J].西南林学院学报,2003,23(2):16-18.
[56]周传明,赖家业,贾勇炯等.厚荚相思离体培养的初步研究[J].四川大学学报(自然科学版),2002,39(3):561-564.
[57]张祖荣,刘兴良.灰木相思茎段腋芽的组织培养及植株再生[J].西南农业大学学报,2004,26(3):291-293.
[58]张祖荣.贝利氏相思的组织培养和快速繁殖[J].西南农业大学学报,2007,29(3):87-89
[59]林来水.黑木相思优良无性系离体培养与快速繁殖技术研究[D].福建农林大学硕士论文,2008,1-73.
[60]苏锦强,张方秋.黑木相思根蘖促萌及组培繁育技术研究[J].广东林业科技, 2008,(03).
[61]杨敏.流苏相思组织培养与再生体系的建立[D] .福建农林大学硕士论文,2008.
[62]李小媛.薄荚相思和流苏相思离体培养与人工种子制作[D] .福建农林大学硕士论文,2008.
[63]王金祥,万小荣.台湾相思的组织培养.华南师范大学学报,2001,03:45-48.
[64]刘娟旭,刘玲,王静,余义勋.大叶相思下胚轴离体培养再生植株的研究[J].林业科学研究,2008,(03):23-34.
[65]潘辉.相思人工林生物量分配格局与林下植被多样性分析[J].福建林业科技, 2008, 1:36-42.
[66]秦武明,何斌,覃世赢.厚荚相思人工林生物量和生产力的研究[J].西北林学院学报, 2008,2,56-58.
[67]陈瑞炳.3种相思类树种生长对比试验[J].山西林业科技, 2008,1,45-56.
[68]覃永华.三个相思树种的造林对比试验[J].广西科技, 2008,1
[69]郭清明.滨海沙地三种相思生长防护效果及改土能力研究[J].海峡科学, 2008, 03.
[70]郑小贤.尾叶桉与马占相思人工复层林生物量及生产力研究叶绍明[J].北京林业大学学报, 2008, 03.
[71]秦武明,唐继新,苏有文,覃永华.厚荚相思人工林综合效益评价[J].安徽农业科学,2008, 14.
[2]王忠.植物生理学[M].北京:中国农业出版,2003:323.
[3] Niu G,Kozai T,Kitaya Y.Simulation of the time course of CO2 concentration inthe culture vessel and net pHotosynthetic rate of Cymbidium planlets[J].Transactions of the ASAE,1996,39(4):1567-1573.
[4]Kozai T,Fujiwara K,Kitaya Y.Modeling measurement and control in plant tissue culture [J].Acta Hort,1993,343:68-78.
[5]Afreen Supporting material affects the growth and development of in vitrosweet potato plantlets cultured pHotoautop Hical1y [J].In Vitro Cell Biol Plant,l999,35:470-476.
[6]王蒂,李友勇.应用生物技术[M].中国农业出版社,1997,23-45.
[7]吴丽君.木木植物组织培养技术在林业科研与生产中的应用与局限[J],福建林业科技,2003, 30(1):67-69.
[8]施寄森.迎接21世纪现代林木生物技术育种的挑战[J].南京林业大学学报,2000,22(1):1-6.
[9]韩一凡.德国林木生物技术研究现状[J].世界林业研究,1993,2: 23-26.
[10]杜克久,郑均宝,徐振华,等.741杨器官、体细胞胚胎发生的细胞组织学及生理学的研究[J].林业科学,1998,34(6):99-104.
[11]FAN Guo-giang,ZHAI Xiao-giao,ZHAI Cui-uan,BI Hui-tao,Callus induction from leaves of different paulownia species and its plantlet regeneration [J].Journal of Forestry Research,2001,12(4):209-214.
[12]范国强,翟晓巧,毕会涛,等.金丝橄的离体培养和植株再生[J].植物生理学通讯,2002,38(4):359.
[13]JAIN S M,GUPTA P K,NEWTON R J.Somatic embryogenesis in woody plants.Vol 1-3,Kluwer Academic Publishers, Dordrecht,The Netherlands.1995:335-341.
[14]赵沛基,彭丽萍,甘烦远,等.刺果番荔枝的组织培养[J].植物生理学通讯,2000,36(2):137-138.
[15]唐巍,欧阳藩,郭仲深.针叶树体细胞无性系研究和应用进展[J].生物工程进展,1997,17(4):2-9.
[16]王仲英,王艺,童德中.生物技术在果树上的应用[J].世界农业,1997,12(24):20-23.
[17]孙清荣等.巴城桃子叶不定植株再生[J].山东农业大学学报,1997,30(4):395-397.
[18]Roche T D,Long R D,and Sayegh A J. Commercial-scale pHotoautotropHic micropropagations in Irish agriculture, horticulture and forestry[J].Acta Hort,1996,440:515-520.
[19]Avila A DE L,Pereyra S M,Arguello J A.Nitrogen Concentration and Proportion of NH4+-N Afect Potato Cultivar Response Media[J].HortScience,1998,33(2):336-338.
[20]李世承,李晶,伶凤琴,等.山杨、大青杨、毛白杨组织培养的研究[J].辽宁大学学报(自然科学版),1995,22(2):59-62.
[21]Attree SM,FowkeL C.Embryogeny of gymnosperms:advances in synthetic see technology of conifers[J].Plant Cell Tiss.Org.Cult., 1993,35:1-35.
[22]王傲雪,李景富.植物体细胞胚状体的诱导研究及应用[J].黑龙江农业科学,1999,2:40-41.
[23]王水,贾勇炯,魏峰,等.云南豆杉的组织培养及植株再生[J].云南植物研究,1997,19(4):407-410.
[24]Michio K,Masakatu O,Michiko A,et a1.The effect of carbon dioxide enrichment,and light in tensity on growth, photosynthesis,and transpiration of carliflower plantlets cultured in vitro pHotoautotropHically and photomixotropHically[J].J Amer Soc HortSci,1998,123(2):176-181.
[25]王秀松,胡东坡等.杜仲愈伤组织的诱导及植株再生的研究[J].西北林学院学报,1994,9(4):32-35.
[26]郑晴霞,王凤翱.绒毛皂英离体茎段培养及再生植株的细胞组织学研究[J].湖南农学院学报,1994,20(3):244-248.
[27]李玲,韩一凡.杨树耐盐突变体筛选的研究[J].林业科学,1990,26(4):359-362.
[28]Y.Desjardins.PHotosynthesis in Vitro on the Factors Regulating CO2 Assimilation in Micropropagation Systems[J].Acta Hortculture,1995(393):45-61.
[29]吴清,闰勇,梁国鲁.杨桃胚乳愈伤组织的诱导和三倍体植株再生[J].热带作物学报,2002,23(2):54-57.
[30]何业华,胡芳名,谢碧霞.经济林木离体培养研究进展[J].中南林学院学报,2000,20(1):31-39.
[31]杜雪梅,李登科.落叶果树生物技术育种研究进展[J].果树学报,2002,(5):286-291.
[32]Seko Y, Nishimura M. Effects of CO2 and light on survival and growth of rice regenerates in vitro on sugar-free medium [J]. Plant cell,tissue and organ culture,1997,46 (3): 257-264.
[33]容世清.桉树无性系湛江6号的工厂化微繁研究初报[J].桉树科技,1995 (2):58-59.
[34]王丽娟,田京伟,杨建雄,等.植物组织培养的研究进展[J].运城高等师范专科学校学报,1999,17(3):8-11.
[35]黄维南.固氮树种组织和细胞培养的种类名录[J].亚热带植物通讯,1998. 27(1):21-25.
[36]施寄森.迎接21世纪现代林木生物技术育种的挑战[J].南京林业大学学报, 2000. 24 (1):1-6.
[37]梁一池,杨华.植物组织培养技术的研究进展[J].福建林学院学报,2002, 22 (1): 93-96.
[38]贾利敏,张红梅,贾利欣.玉米组织培养和植株再生的研究现状[J].内蒙古农业科技, 2002,12(3):10-13.
[39]祁永斌,李和平.不同小麦品种愈伤组织诱导和再生体系建立[J].武汉植物学研究,2005,23(3):227-232.
[40]李雪,韩烈保.多年生黑麦草愈伤组织诱导与芽分化的研究[J].草业科学,2005,22(5):45-47.
[41]吴桂胜,胡鸢雷等.剪股颖愈伤组织诱导与植株再生[J].生物技术通报,2006,(4):101-102.
[42]谢启鑫.君迁子叶片培养再生植株的研究[J].中国农业科学, 2008,41(2):607-612.
[43]苗永美.菘蓝两种外植体愈伤组织诱导[J].生物科技通报, 2008, 4(2):45-48.
[44]苏明申,林顺权.荔枝愈伤组织诱导及其体细胞胚胎发生研究初报[J].华南热带农业大学学报,2004,15(9):26-28.
[45]范国强,黎明.悬铃木体细胞胚胎发生及植株再生[J].林业科学,2004,40(3):28-29.
[46]于志水,张志毅.杨树体细胞胚胎发生研究概况[J].东北林业大学学报,2005,33(1):21-23.
[47]姬飞腾,李凤兰.香果树体细胞胚胎发生[J].植物生理学通讯,2005,41(5):19-21.
[48]Attree.S.M,Fowke.L.C. Embryogency of gymnosperms.Plant Cell Tiss Organ Cult,1993,35:1-35.
[49]Jain.S. M, Gupta .P. K,Newton .R. J.Somatic embryogenesis in woody plants. Vol 1-3, Kluwer Academic Publishers, Dordrecht,The Netherlands.1995:335-341.
[50]Tautorus .J .E, Fowke .L .C,Dunstand .I.Somatic embryogenesis in conifers.Candian Joural of Bocany,1991,69:1873-1899
[51]张存旭.栓皮栎体细胞发生的细胞组织学观察[J].植物生理和分子生物学学报, 2007, 33(1) :33-38.
[52]萧洪东.红花银桦组织培养与快速繁殖[J].植物生理学通报, 2008,44(01):34-41.
[53]张宏伟,黄学林,傅家瑞,等.大叶相思、马占相思腋芽培养和植株再生[J].热带亚热带植物学报,1995,3(3):62-68.
[54]蔡玲,王以红,吴幼媚,等.马尖相思离体培养再生植株的研究[J].广西林业科学,1999,03: 258-261.
[55]高洁,玉玲.大叶相思茎段腋芽组织培养技术初探[J].西南林学院学报,2003,23(2):16-18.
[56]周传明,赖家业,贾勇炯等.厚荚相思离体培养的初步研究[J].四川大学学报(自然科学版),2002,39(3):561-564.
[57]张祖荣,刘兴良.灰木相思茎段腋芽的组织培养及植株再生[J].西南农业大学学报,2004,26(3):291-293.
[58]张祖荣.贝利氏相思的组织培养和快速繁殖[J].西南农业大学学报,2007,29(3):87-89
[59]林来水.黑木相思优良无性系离体培养与快速繁殖技术研究[D].福建农林大学硕士论文,2008,1-73.
[60]苏锦强,张方秋.黑木相思根蘖促萌及组培繁育技术研究[J].广东林业科技, 2008,(03).
[61]杨敏.流苏相思组织培养与再生体系的建立[D] .福建农林大学硕士论文,2008.
[62]李小媛.薄荚相思和流苏相思离体培养与人工种子制作[D] .福建农林大学硕士论文,2008.
[63]王金祥,万小荣.台湾相思的组织培养.华南师范大学学报,2001,03:45-48.
[64]刘娟旭,刘玲,王静,余义勋.大叶相思下胚轴离体培养再生植株的研究[J].林业科学研究,2008,(03):23-34.
[65]潘辉.相思人工林生物量分配格局与林下植被多样性分析[J].福建林业科技, 2008, 1:36-42.
[66]秦武明,何斌,覃世赢.厚荚相思人工林生物量和生产力的研究[J].西北林学院学报, 2008,2,56-58.
[67]陈瑞炳.3种相思类树种生长对比试验[J].山西林业科技, 2008,1,45-56.
[68]覃永华.三个相思树种的造林对比试验[J].广西科技, 2008,1
[69]郭清明.滨海沙地三种相思生长防护效果及改土能力研究[J].海峡科学, 2008, 03.
[70]郑小贤.尾叶桉与马占相思人工复层林生物量及生产力研究叶绍明[J].北京林业大学学报, 2008, 03.
[71]秦武明,唐继新,苏有文,覃永华.厚荚相思人工林综合效益评价[J].安徽农业科学,2008, 14.