流苏相思优良无性系离体培养与快速繁殖技术研究
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摘要
流苏相思(Acacia fimbriata),为含羞草科(Mimosaceae)金合欢属(Acacia)植物,它根瘤发达,固氮能力强,且生物量大,枯枝落叶多,对土壤有很好的保护和改良作用。随着我国园林事业的发展,这种具有生态价值又具有显著观赏价值的观赏相思树,必然会引起人们更大的开发和研究兴趣。本论文以流苏相思优良无性系茎段为基本材料,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进行研究,目的在于探索流苏相思优良无性系离体培养与快速繁殖的有效途径,以便建立快速高效的流苏相思组织培养技术体系,为其工厂化生产提供了一定的理论基础和技术支持,研究结果表明:
1、流苏相思外植体经洗衣粉水浸泡20 min,自来水冲洗25 min,以75%乙醇浸泡20s,0.1%升汞10min,无菌水冲洗5遍,污染率仅为26.7%。
2、流苏相思优良无性系离体培养适宜的外植体类型为新枝的上部带腋芽的茎段,其污染率可控制在30%以内、一周左右即可分化出芽;适宜流苏相思不定芽分化成苗的季节为秋天;适宜流苏相思不定芽分化的培养基MS+BA1.0mg/L +KT1.0mg/L +IBA1.0mg/L +蔗糖30 g /L,分化率达到79.2%。
3.流苏相思增殖培养。采用正交试验设计,考虑不同植物激素对流苏相思增殖的影响,筛选出一代培养基为:MS+BA1.0mg/L +KT1.0 mg/L +IBA1.0mg/L +蔗糖30g/L,30d增殖数达到3.36;流苏相思幼苗在二次继代中最好的培养基为:MS+KT0.4mg/L +NAA0.1mg /L +蔗糖20g/L,增殖数达到3.03。在流苏相思的三代及多代培养中,选择1/2MS+ KT0.2mg /L +IBA0.2 mg/L +活性碳1g/L +蔗糖15g/L的培养基,增殖效果比较理想,增殖数达到了3.03。
4.流苏相思的壮苗及生根。第三次继代培养基可以起到壮苗的效果。根据其生根情况,筛选出较佳的培养基配方为:1/2MS +IAA0.2mg/L +IBA0.3mg/L +NAA0.1mg/L +蔗糖15g /L,生根率达到了88%。
5.流苏相思的移栽。移栽基质选择珍珠岩:草炭土=2:1,移栽成活率可以达到80%,植株生长健壮。炼苗天数以7～9d为宜。
Acacia fimbriata belongs to Mimosaceae family and Acacia genera. It has well developed root nodule, strong ability of nitrogen fixation, high biomass and multiple litter, which has very good protection and modified effect on soil. With the development of our country park business, the ornamental acacia which has ecological value and significant value will necessarily attract more people’s developing and studying interest. This paper basic materials for Acacia fimbriata elite stem, sterile system from the establishment subculture, rooting induction hardened various links transplant research aimed at exploring Acacia fimbriata elite in vitro culture and rapid propagation of the effective ways to build fast, efficient Acacia fimbriata tissue culture technology system. Industrial production for providing a theoretical basis and technological support. the findings show that:
1. The explants of Acacia fimbriata immersed in detergent for 20 minutes ,then was washed with water for 25minutes. The next disinfection was soaking for twenty second with 75% alcohol, then sterilizing in 0.1% mercuric for ten minutes, finally washing with sterile water for five times. The infected rate was only 26.7%.
2. The suitable explant of tissue culture of Acacia fimbriata was the top of the new stem with axillary buds . It will differentiate in one week and its pollution rate not more than 30 percent.The season of autumn could promote the differentiation of adventitious buds.The culture medium MS+IBA1.0mg/L +KT1.0mg/L +IBA1.0mg/L +Sucrose30g/Lcould promote the differentiation of adventitious buds, Differentiation rate is 79.2percent.
3. The method of orthogonal design was adopted to arrange experiment. Considering the influence of plant hormone on the proliferation culture of Acacia fimbriata, selected MS+BA1.0mg /L +KT1.0 mg/L +IBA1.0mg/L + Sucrose 30g/L as the first culture medium, the proliferation coefficient was 3.36. The highest proliferation coefficient of the Acacia fimbriata in the second multiplication culture, was 3.03 with the medium combination of MS+KT0.4mg/L +NAA0.1mg /L + Sucrose 20g/L,. During the third and multi-generation multiplication culture, taking the 1/2MS+KT0.2mg/L +IBA0.2mg/L +AC1g/L + Sucrose 15g/L as the medium had a better multiplication effect with the proliferation coefficient 3.03.
4.The strong seedling and root culture of Acacia fimbriata.The strong seedling medium was the medium of the third multiplication culture. According to the rooting situation, the better medium was 1/2MS +IAA0.2mg/L +IBA0.3mg/L +NAA0.1mg/L + Sucrose 15g/L,with the rooting rate 88%.
5.The transplanting of Acacia fimbriata.Combination of and peat soil with a ratio of 2:1 was used for transplanting, and the survival rate was 80%. The plant was strong.The suitable acclimatization time were 7～9 days.
1、流苏相思外植体经洗衣粉水浸泡20 min,自来水冲洗25 min,以75%乙醇浸泡20s,0.1%升汞10min,无菌水冲洗5遍,污染率仅为26.7%。
2、流苏相思优良无性系离体培养适宜的外植体类型为新枝的上部带腋芽的茎段,其污染率可控制在30%以内、一周左右即可分化出芽;适宜流苏相思不定芽分化成苗的季节为秋天;适宜流苏相思不定芽分化的培养基MS+BA1.0mg/L +KT1.0mg/L +IBA1.0mg/L +蔗糖30 g /L,分化率达到79.2%。
3.流苏相思增殖培养。采用正交试验设计,考虑不同植物激素对流苏相思增殖的影响,筛选出一代培养基为:MS+BA1.0mg/L +KT1.0 mg/L +IBA1.0mg/L +蔗糖30g/L,30d增殖数达到3.36;流苏相思幼苗在二次继代中最好的培养基为:MS+KT0.4mg/L +NAA0.1mg /L +蔗糖20g/L,增殖数达到3.03。在流苏相思的三代及多代培养中,选择1/2MS+ KT0.2mg /L +IBA0.2 mg/L +活性碳1g/L +蔗糖15g/L的培养基,增殖效果比较理想,增殖数达到了3.03。
4.流苏相思的壮苗及生根。第三次继代培养基可以起到壮苗的效果。根据其生根情况,筛选出较佳的培养基配方为:1/2MS +IAA0.2mg/L +IBA0.3mg/L +NAA0.1mg/L +蔗糖15g /L,生根率达到了88%。
5.流苏相思的移栽。移栽基质选择珍珠岩:草炭土=2:1,移栽成活率可以达到80%,植株生长健壮。炼苗天数以7～9d为宜。
Acacia fimbriata belongs to Mimosaceae family and Acacia genera. It has well developed root nodule, strong ability of nitrogen fixation, high biomass and multiple litter, which has very good protection and modified effect on soil. With the development of our country park business, the ornamental acacia which has ecological value and significant value will necessarily attract more people’s developing and studying interest. This paper basic materials for Acacia fimbriata elite stem, sterile system from the establishment subculture, rooting induction hardened various links transplant research aimed at exploring Acacia fimbriata elite in vitro culture and rapid propagation of the effective ways to build fast, efficient Acacia fimbriata tissue culture technology system. Industrial production for providing a theoretical basis and technological support. the findings show that:
1. The explants of Acacia fimbriata immersed in detergent for 20 minutes ,then was washed with water for 25minutes. The next disinfection was soaking for twenty second with 75% alcohol, then sterilizing in 0.1% mercuric for ten minutes, finally washing with sterile water for five times. The infected rate was only 26.7%.
2. The suitable explant of tissue culture of Acacia fimbriata was the top of the new stem with axillary buds . It will differentiate in one week and its pollution rate not more than 30 percent.The season of autumn could promote the differentiation of adventitious buds.The culture medium MS+IBA1.0mg/L +KT1.0mg/L +IBA1.0mg/L +Sucrose30g/Lcould promote the differentiation of adventitious buds, Differentiation rate is 79.2percent.
3. The method of orthogonal design was adopted to arrange experiment. Considering the influence of plant hormone on the proliferation culture of Acacia fimbriata, selected MS+BA1.0mg /L +KT1.0 mg/L +IBA1.0mg/L + Sucrose 30g/L as the first culture medium, the proliferation coefficient was 3.36. The highest proliferation coefficient of the Acacia fimbriata in the second multiplication culture, was 3.03 with the medium combination of MS+KT0.4mg/L +NAA0.1mg /L + Sucrose 20g/L,. During the third and multi-generation multiplication culture, taking the 1/2MS+KT0.2mg/L +IBA0.2mg/L +AC1g/L + Sucrose 15g/L as the medium had a better multiplication effect with the proliferation coefficient 3.03.
4.The strong seedling and root culture of Acacia fimbriata.The strong seedling medium was the medium of the third multiplication culture. According to the rooting situation, the better medium was 1/2MS +IAA0.2mg/L +IBA0.3mg/L +NAA0.1mg/L + Sucrose 15g/L,with the rooting rate 88%.
5.The transplanting of Acacia fimbriata.Combination of and peat soil with a ratio of 2:1 was used for transplanting, and the survival rate was 80%. The plant was strong.The suitable acclimatization time were 7～9 days.
引文
[1]周鸿彬,高本旺,高登梅,等.相思类树种引种初报[J ].湖北林业科技, 2001 ,4) : 19-22.
[2]李纪元.金合欢属植物资源在我国亚热带的引种潜力[J ].福建林学院学报,2002,22(3) : 283 - 288.
[3]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 (4) : 16 - 20.
[4]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(l):67-69.
[5]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.
[6]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.
[7]胡含,王恒力.植物细胞工程与育种[M].北京:北京工业大学出版社,1990,171-185.
[8]王纪方,金波,高秀云,等[M].上海:上海科学技术出版社,1983.
[9]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,(3):10-13
[10]李雪,韩烈保,等.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
[11]祁永斌,李和平,等.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
[12]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
[13]沙红,唐芳,高燕,张瑞麟.大红红景天愈伤组织的诱导及植株再生体系的建立[J].新疆农业大学学报,2008,31(3):58-61
[14]马艳,闵义,陈小文,董学会.罗布麻愈伤组织诱导及植株再生[J].西北植物学报,2008 ,28 (8) :1580 - 1585
[15]施波,孙小琴,任甜甜,石开明,丁莉,周毅峰,周光来.青蒿茎段愈伤组织的诱导及植株再生[J].湖北民族学院学报,2008 ,26 (2) :217- 219
[16]范国强,翟晓巧,李松林.泡桐愈伤组织再生植株的诱导与培养[J].植物学通报,2002,19(1):92-97
[17]康冰,王关平,陈彦生.速生欧美黑杨愈伤组织诱导及植株再生[J].植物生理学通讯, 2004,40(5):582-584
[18]曾建军,肖宜安,孙敏.长柄双花木愈伤组织诱导及植株再生[J].广西植物,2006,26 ( 6) : 628 - 630
[19]Attree.S.M,Fowke.L.C.Embryogencyofgymnosperms.Plant Cell Tiss Organ Cult,1993,35: 1-35
[20]Jain.S. M,Gupta .P. K,Newton .R. J.Somatic embryogenesis in woody plants. Vol1-3, Kluwer Academic Publishers,Dordrecht,The Netherlands.1995:335-341
[21]Tautorus .J .E,Fowke .L .C,Dunstand .I.Somatic embryogenesis in conifers.Candian Joural of Bocany,1991,69:1873-1899
[22]沈海龙.植物组织培养[M].中国农业出版社,2005.
[23]陈金慧,施季森,诸葛强,等.植物体细胞胚胎发生机理的研究进展.南京林业大学学报,2003,27(1):75_80.
[24]张存旭,姚增玉,等.栓皮栎体胚诱导关键影响因素研究.林业科学,2005,41(2):174-177
[25]陈颖,曹福亮.银杏组培快繁和体胚发生技术研究进展.林业科技开发,2006,20(6):10-12
[26]莫小路,曾庆钱,邱蔚芬,陈瑜珍.檀香体细胞胚胎的发生及植株再生的研究[J].食品与药品, 2008,10 ( 01) : 35 -37
[27]闫红舟.植物体细胞胚胎发生的研究进展[J].河北农业科学, 2008, 12 (3) : 4 - 5
[28]赵泊,程井辰.石刁柏体细胞胚发生的组织学研究[J].植物学报,1994,34 C7): 491-495.
[29]欧阳杰,武彦文,王晓东,赵兵,王玉春.肉苁蓉细胞悬浮培养体系的建立[J].中草药, 2008, 39 (03) :426 - 428
[30]周晓鹿,李思良.皂质芦荟细胞悬浮培养初探[J].北方园艺,2008,(2) :188-192
[31]丁婷,马庆,项艳.红景天愈伤组织的诱导及细胞悬浮培养[J].中国农学通报,2008, 24(5) : 224-227
[32]雷泽勇,周风艳,周莉.关于离体胚组织培养在植物育种中的应用进展[J].防护林科技,2001(3):57-59.
[33]Afreen Supporting material affects the growth and development of in vitrosweet potato plantlets cultured photoautophical1y[J].In Vitro Cell Biol Plant,l999,35:470-474.
[34]赖钟雄,陈振光.龙眼胚性细胞悬浮培养再生植株.应用与环境生物学报,2002(5):27-29.
[35]唐立俊.我国果树原生质体培养研究[J].遵义师范学院学报, 2007, 9(2) :66-69
[36]彭爱红,何永謇,邹修早,等.观赏植物组织培养与基因工程研究进展(综述)[J].亚热带植物科学,2002,31(2):58-63.
[37]丁燕芬,王岩,龙春林.中国兰花组织培养研究进展[J].安徽农业科学, 2007, (8):47-48.
[38]王笑春,李峰,蒋湘宁.桉树组织培养与再生系统建立研究[J].河北建筑科技学院学报, 2006,(2):71-74.
[39]刘欣,薛金芝,赵丹,裴崎君.植物组织培养技术在林木育种中的应用[J].吉林林业科技, 2007,(3):8-10.
[40]杨斌,张超,兰天维,等.羽叶蔓绿绒的组织培养和快速繁殖[J].植物生理学通讯,2008,(1).
[41]陈春满,蒋雄辉,郑贵朝,等.白叶瓶子草的组织培养和快速繁殖[J].植物生理学通讯,2008, (1):27.
[42]吴幼媚,陈晓明,王以红,等.大叶栎的组织培养和快速繁殖[J].植物生理学通讯,2008, (01):28.
[43]王方正,蒋雄辉,郑健临.也门铁的组织培养与快速繁殖[J].植物生理学通讯,2006,(3):481 .
[44]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.
[45]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.
[46]周丽华,吕华飞.茎尖分生组织培养获得无病毒康乃馨植株的研究[J].云南农业大学学报,1994, (4):254-256.
[47]孟志卿.草莓花药组织培养脱毒快繁技术研究[J].安徽农业科学,2008,(2):42 .
[48]赵庆芳,马平霞.百合脱毒及病毒鉴定的研究进展[J].北方园艺,2007,(3):77-79.
[49]闫承璞.桃离体培养及叶片再生体系的建立[D].中国农业科学院, 2006.
[50]王鸿.杏的再生体系建立研究[D].西北农林科技大学,2005.
[51]胡军荣.菊花的脱毒与快繁技术[J].安徽农学通报, 2008, 14(5) :142-143
[52]李明军,张晓丽,杜琳,陈建军,赵喜亭,刘杰,周娜,涂荣涛,郭丹.怀地黄试管苗脱毒技术研究[J].河南师范大学学报, 2008, (3) :103-106
[53]孙必贤,朴成学,赵海锋,建德锋.大蒜的脱毒与快繁技术[J].吉林蔬菜, 2008, 9(2) :17-18
[54]江洪如,余发新,刘腾云,朱祺,高柱,王碧琴,周华,王小玲.金边瑞香茎尖脱毒及快繁技[J].南昌大学学报, 2008, 32(1) :59-61
[55]陈志林,李开绵.植物种质资源的超低温保存[J].广西农业科学,2007(3):14-16
[56]谢响明,杨明嘉,尚宇,等.相思树种的外植体繁殖及基因工程研究进展[J].北京林业大学学报,2006,28(4):148～154.
[57]赖家业,周传明,叶春生,等.厚荚相思组织培养与快速繁殖[J].四川大学学报(自然科学版),2003,40(5):983-985.
[58]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(3): 258-261.
[59]苏秀城.黑木相思组织培养的初步研究[J].福建林业科技,1999,26(4):78-81.
[60]王金祥,万小荣,等.台湾相思的组织培养[J].华南师范大学学报,2001(3):45-48.
[61]蔡玲,王以红,等.五种相思树组织培养研究[J].广西林业科学,2003,32(1):24-26.
[62]高洁,玉玲.大叶相思茎段腋芽组织培养技术初探[J].西南林学报,2003,23(2):16-18.
[63]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(自然科学版),2004,26(3):291-293.
[64]张月娇.厚荚相思组培育苗试验[J].福建林业科技,2003,30(4):72-74.
[65]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(自然科学版),2004,26(3):291-293.
[66]裘珍飞,曾炳山,李湘阳,等.马占相思优树组培苗扦插技术研究[J].广东林业科技,2005, 21(2):22-25.
[67]马钟丽.厚荚相思离体快繁技术的研究[D].福建农林大学硕士论文,2005:1-57.
[68]姬明.黑木相思离体培养与再生体系的建立[D].福建农林大学硕士论文,2006:1-44.
[69]李宋香,赖钟雄,林珊珊,陈义挺,等.薄荚相思种子离体萌发与快速繁殖[J].亚热带农业研究,2006,2(4):313-317.
[70]张祖荣.贝利氏相思的组织培养和快速繁殖[J].西南大学学报,2007,29(3):86-89.
[71]黄骐.卷荚相思优树无性系快速繁殖技术[J].林业科技开发,2007,21(5):66-68.
[72]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,2008:1-66.
[73]张祖荣.多花相思的组织培养和快速繁殖[J].福建林业科技,2007,34(2):143-145.
[74]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,2008:1-41.
[75]欧阳彤.姆那格葡萄组培快速繁育[J].科技论坛,1998,27-28.
[76]周飞雪,周俊勤.葡萄组织培养[J].湖南林业,2001(3):19.
[77]袁丽钗.核桃品种试管苗生根及不定根发生过程中特异蛋白表达的研究[D].保定:河北农业大学园艺学院,2000.
[78]丁亚军,代汉萍,李宝江.植物激素和生长调节剂在果树组织培养中的应用[J].北方园艺,2002(6):68-70.
[79]李永红,李艳平.浅谈植物组织培养技术在苗木繁殖中的应用[J].山西水土保持科技,2000(2):25-26.
[80]Leifer C Morris C and Waites WM.Ecology of microbial saprophytes and pathogens in tissueculture and Geld-grown plants:reasons for contamination problems in vitro[J].CRC Crit Rev,Plant Science, 1999(13):139-183.
[81]王玉国.枣树的组织培养与快速繁殖[J].山西农业大学学报,1994,14(4):371-372.
[82]Yan R.L. et al. Effect of exogenous plant hormones on rapid multiplication of ziziphus jujube mill test-tube plants[J].Journal of Fruit Science.1990,7(4):231-233.
[83]金建平.观赏获的组织培养[J].植物生理学通讯.1991,27(1):71-73.
[84]王冬梅.细胞分裂素类物质在植物组织培养中的应用机制[J].植物生理学通讯,1996,32 (5):373-377.
[85]王忠.植物生理学[M].中国农业出版社.2000,323-330.
[86]段新玲,任东岁.毛刺槐的组织培养和植株再生[J].植物生理学通讯,2000, (6): 434-435.
[87]Wong WC. In Vtro propagation of bananas (Musaspp.):Initiation,proliferation and development of shoot tip culture on defined media. Plant Cell[J].Tissue and Organ Culture.1986,6(2):159.
[88] Loescher WH. et al[J].Physiologia Plantanum.1979,47:250.
[89]熊丽,吴丽芳.观赏花卉的组织培养与大规模生产[M].北京:化学工业出版社,2003.
[90]叶叙丰.拟石莲花离体叶片的不定芽分化及其和内源激素状况的关系[J].植物生理学通讯,1989, (4):26-30.
[91]叶秀遴.石斛兰组织培养和细胞学观察[J].园艺学报,1995,22 (1):83-87.
[92]邹翠霞,肖容俊.雪柳的组织培养与植株再生[J].植物生理学通讯,2000,36 (2):136-137.
[93]郭春华,徐玉霞.沙棘优良品系茎尖组织培养技术初探[J].沙棘,2000,13 (1):26-27.
[2]李纪元.金合欢属植物资源在我国亚热带的引种潜力[J ].福建林学院学报,2002,22(3) : 283 - 288.
[3]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 (4) : 16 - 20.
[4]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(l):67-69.
[5]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.
[6]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.
[7]胡含,王恒力.植物细胞工程与育种[M].北京:北京工业大学出版社,1990,171-185.
[8]王纪方,金波,高秀云,等[M].上海:上海科学技术出版社,1983.
[9]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,(3):10-13
[10]李雪,韩烈保,等.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
[11]祁永斌,李和平,等.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
[12]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
[13]沙红,唐芳,高燕,张瑞麟.大红红景天愈伤组织的诱导及植株再生体系的建立[J].新疆农业大学学报,2008,31(3):58-61
[14]马艳,闵义,陈小文,董学会.罗布麻愈伤组织诱导及植株再生[J].西北植物学报,2008 ,28 (8) :1580 - 1585
[15]施波,孙小琴,任甜甜,石开明,丁莉,周毅峰,周光来.青蒿茎段愈伤组织的诱导及植株再生[J].湖北民族学院学报,2008 ,26 (2) :217- 219
[16]范国强,翟晓巧,李松林.泡桐愈伤组织再生植株的诱导与培养[J].植物学通报,2002,19(1):92-97
[17]康冰,王关平,陈彦生.速生欧美黑杨愈伤组织诱导及植株再生[J].植物生理学通讯, 2004,40(5):582-584
[18]曾建军,肖宜安,孙敏.长柄双花木愈伤组织诱导及植株再生[J].广西植物,2006,26 ( 6) : 628 - 630
[19]Attree.S.M,Fowke.L.C.Embryogencyofgymnosperms.Plant Cell Tiss Organ Cult,1993,35: 1-35
[20]Jain.S. M,Gupta .P. K,Newton .R. J.Somatic embryogenesis in woody plants. Vol1-3, Kluwer Academic Publishers,Dordrecht,The Netherlands.1995:335-341
[21]Tautorus .J .E,Fowke .L .C,Dunstand .I.Somatic embryogenesis in conifers.Candian Joural of Bocany,1991,69:1873-1899
[22]沈海龙.植物组织培养[M].中国农业出版社,2005.
[23]陈金慧,施季森,诸葛强,等.植物体细胞胚胎发生机理的研究进展.南京林业大学学报,2003,27(1):75_80.
[24]张存旭,姚增玉,等.栓皮栎体胚诱导关键影响因素研究.林业科学,2005,41(2):174-177
[25]陈颖,曹福亮.银杏组培快繁和体胚发生技术研究进展.林业科技开发,2006,20(6):10-12
[26]莫小路,曾庆钱,邱蔚芬,陈瑜珍.檀香体细胞胚胎的发生及植株再生的研究[J].食品与药品, 2008,10 ( 01) : 35 -37
[27]闫红舟.植物体细胞胚胎发生的研究进展[J].河北农业科学, 2008, 12 (3) : 4 - 5
[28]赵泊,程井辰.石刁柏体细胞胚发生的组织学研究[J].植物学报,1994,34 C7): 491-495.
[29]欧阳杰,武彦文,王晓东,赵兵,王玉春.肉苁蓉细胞悬浮培养体系的建立[J].中草药, 2008, 39 (03) :426 - 428
[30]周晓鹿,李思良.皂质芦荟细胞悬浮培养初探[J].北方园艺,2008,(2) :188-192
[31]丁婷,马庆,项艳.红景天愈伤组织的诱导及细胞悬浮培养[J].中国农学通报,2008, 24(5) : 224-227
[32]雷泽勇,周风艳,周莉.关于离体胚组织培养在植物育种中的应用进展[J].防护林科技,2001(3):57-59.
[33]Afreen Supporting material affects the growth and development of in vitrosweet potato plantlets cultured photoautophical1y[J].In Vitro Cell Biol Plant,l999,35:470-474.
[34]赖钟雄,陈振光.龙眼胚性细胞悬浮培养再生植株.应用与环境生物学报,2002(5):27-29.
[35]唐立俊.我国果树原生质体培养研究[J].遵义师范学院学报, 2007, 9(2) :66-69
[36]彭爱红,何永謇,邹修早,等.观赏植物组织培养与基因工程研究进展(综述)[J].亚热带植物科学,2002,31(2):58-63.
[37]丁燕芬,王岩,龙春林.中国兰花组织培养研究进展[J].安徽农业科学, 2007, (8):47-48.
[38]王笑春,李峰,蒋湘宁.桉树组织培养与再生系统建立研究[J].河北建筑科技学院学报, 2006,(2):71-74.
[39]刘欣,薛金芝,赵丹,裴崎君.植物组织培养技术在林木育种中的应用[J].吉林林业科技, 2007,(3):8-10.
[40]杨斌,张超,兰天维,等.羽叶蔓绿绒的组织培养和快速繁殖[J].植物生理学通讯,2008,(1).
[41]陈春满,蒋雄辉,郑贵朝,等.白叶瓶子草的组织培养和快速繁殖[J].植物生理学通讯,2008, (1):27.
[42]吴幼媚,陈晓明,王以红,等.大叶栎的组织培养和快速繁殖[J].植物生理学通讯,2008, (01):28.
[43]王方正,蒋雄辉,郑健临.也门铁的组织培养与快速繁殖[J].植物生理学通讯,2006,(3):481 .
[44]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.
[45]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.
[46]周丽华,吕华飞.茎尖分生组织培养获得无病毒康乃馨植株的研究[J].云南农业大学学报,1994, (4):254-256.
[47]孟志卿.草莓花药组织培养脱毒快繁技术研究[J].安徽农业科学,2008,(2):42 .
[48]赵庆芳,马平霞.百合脱毒及病毒鉴定的研究进展[J].北方园艺,2007,(3):77-79.
[49]闫承璞.桃离体培养及叶片再生体系的建立[D].中国农业科学院, 2006.
[50]王鸿.杏的再生体系建立研究[D].西北农林科技大学,2005.
[51]胡军荣.菊花的脱毒与快繁技术[J].安徽农学通报, 2008, 14(5) :142-143
[52]李明军,张晓丽,杜琳,陈建军,赵喜亭,刘杰,周娜,涂荣涛,郭丹.怀地黄试管苗脱毒技术研究[J].河南师范大学学报, 2008, (3) :103-106
[53]孙必贤,朴成学,赵海锋,建德锋.大蒜的脱毒与快繁技术[J].吉林蔬菜, 2008, 9(2) :17-18
[54]江洪如,余发新,刘腾云,朱祺,高柱,王碧琴,周华,王小玲.金边瑞香茎尖脱毒及快繁技[J].南昌大学学报, 2008, 32(1) :59-61
[55]陈志林,李开绵.植物种质资源的超低温保存[J].广西农业科学,2007(3):14-16
[56]谢响明,杨明嘉,尚宇,等.相思树种的外植体繁殖及基因工程研究进展[J].北京林业大学学报,2006,28(4):148～154.
[57]赖家业,周传明,叶春生,等.厚荚相思组织培养与快速繁殖[J].四川大学学报(自然科学版),2003,40(5):983-985.
[58]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(3): 258-261.
[59]苏秀城.黑木相思组织培养的初步研究[J].福建林业科技,1999,26(4):78-81.
[60]王金祥,万小荣,等.台湾相思的组织培养[J].华南师范大学学报,2001(3):45-48.
[61]蔡玲,王以红,等.五种相思树组织培养研究[J].广西林业科学,2003,32(1):24-26.
[62]高洁,玉玲.大叶相思茎段腋芽组织培养技术初探[J].西南林学报,2003,23(2):16-18.
[63]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(自然科学版),2004,26(3):291-293.
[64]张月娇.厚荚相思组培育苗试验[J].福建林业科技,2003,30(4):72-74.
[65]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(自然科学版),2004,26(3):291-293.
[66]裘珍飞,曾炳山,李湘阳,等.马占相思优树组培苗扦插技术研究[J].广东林业科技,2005, 21(2):22-25.
[67]马钟丽.厚荚相思离体快繁技术的研究[D].福建农林大学硕士论文,2005:1-57.
[68]姬明.黑木相思离体培养与再生体系的建立[D].福建农林大学硕士论文,2006:1-44.
[69]李宋香,赖钟雄,林珊珊,陈义挺,等.薄荚相思种子离体萌发与快速繁殖[J].亚热带农业研究,2006,2(4):313-317.
[70]张祖荣.贝利氏相思的组织培养和快速繁殖[J].西南大学学报,2007,29(3):86-89.
[71]黄骐.卷荚相思优树无性系快速繁殖技术[J].林业科技开发,2007,21(5):66-68.
[72]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,2008:1-66.
[73]张祖荣.多花相思的组织培养和快速繁殖[J].福建林业科技,2007,34(2):143-145.
[74]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,2008:1-41.
[75]欧阳彤.姆那格葡萄组培快速繁育[J].科技论坛,1998,27-28.
[76]周飞雪,周俊勤.葡萄组织培养[J].湖南林业,2001(3):19.
[77]袁丽钗.核桃品种试管苗生根及不定根发生过程中特异蛋白表达的研究[D].保定:河北农业大学园艺学院,2000.
[78]丁亚军,代汉萍,李宝江.植物激素和生长调节剂在果树组织培养中的应用[J].北方园艺,2002(6):68-70.
[79]李永红,李艳平.浅谈植物组织培养技术在苗木繁殖中的应用[J].山西水土保持科技,2000(2):25-26.
[80]Leifer C Morris C and Waites WM.Ecology of microbial saprophytes and pathogens in tissueculture and Geld-grown plants:reasons for contamination problems in vitro[J].CRC Crit Rev,Plant Science, 1999(13):139-183.
[81]王玉国.枣树的组织培养与快速繁殖[J].山西农业大学学报,1994,14(4):371-372.
[82]Yan R.L. et al. Effect of exogenous plant hormones on rapid multiplication of ziziphus jujube mill test-tube plants[J].Journal of Fruit Science.1990,7(4):231-233.
[83]金建平.观赏获的组织培养[J].植物生理学通讯.1991,27(1):71-73.
[84]王冬梅.细胞分裂素类物质在植物组织培养中的应用机制[J].植物生理学通讯,1996,32 (5):373-377.
[85]王忠.植物生理学[M].中国农业出版社.2000,323-330.
[86]段新玲,任东岁.毛刺槐的组织培养和植株再生[J].植物生理学通讯,2000, (6): 434-435.
[87]Wong WC. In Vtro propagation of bananas (Musaspp.):Initiation,proliferation and development of shoot tip culture on defined media. Plant Cell[J].Tissue and Organ Culture.1986,6(2):159.
[88] Loescher WH. et al[J].Physiologia Plantanum.1979,47:250.
[89]熊丽,吴丽芳.观赏花卉的组织培养与大规模生产[M].北京:化学工业出版社,2003.
[90]叶叙丰.拟石莲花离体叶片的不定芽分化及其和内源激素状况的关系[J].植物生理学通讯,1989, (4):26-30.
[91]叶秀遴.石斛兰组织培养和细胞学观察[J].园艺学报,1995,22 (1):83-87.
[92]邹翠霞,肖容俊.雪柳的组织培养与植株再生[J].植物生理学通讯,2000,36 (2):136-137.
[93]郭春华,徐玉霞.沙棘优良品系茎尖组织培养技术初探[J].沙棘,2000,13 (1):26-27.