澳大利亚特色花卉Sun Fan无性繁殖研究
|
摘要
Sun Fan,Scaevola属中的一种,拉丁文名(Scaevola aemula),多年生的草本植物,为澳大利亚特产,适应性强,花形特别、颜色鲜艳,自1989年进入花卉市场后,很快就成为澳大利亚花卉市场的重要种类,并已在欧洲和北美引种成功,深受消费者喜爱,被广泛用作吊篮、地栽花卉、盆栽花卉及鲜切花。我国尚无此类花卉,因此Sun Fan是极具开发价值的花卉。由于传统播种育苗、分株繁殖所获得的苗木数量有限,满足不了市场的需求,基于这种情况,本论文开展了Sun Fan组织培养和扦插繁殖技术研究,以期摸索出提高Sun Fan繁殖数量和质量的有效途径,为苗木产业化生产、优良种质保存提供理论和技术支撑。
采用组织培养技术手段,探讨了从无菌体系的建立、愈伤诱导、不定芽分化、增殖培养、生根培养到试管苗移栽的整个过程,经过研究分析,得出以下结论:
1.以Sun Fan嫩叶、老叶、茎段、茎节为外植体,消毒方式为:75%酒精浸泡30s,0.1%HgCl_2消毒3min,接种在MS基本培养基,附加0.1mg·L~(-1)NAA进行培养,试验结果表明,嫩叶愈伤组织诱导率最高,达87.5%;嫩叶为Sun Fan最佳组织培养外植体。
2.愈伤诱导以Sun Fan嫩叶为外植体,以MS为基本培养基,L_(16)(4~3)正交试验设计,统计分析结果表明,6-BA、2,4-D、NAA对Sun Fan嫩叶愈伤组织的诱导影响均达到极显著水平,最佳愈伤诱导培养基为:MS+6-BA0.50mg·L~(-1)+2,4-D0.20mg·L~(-1)+NAA0.10mg·L~(-1),诱导率高达100%。
3.不定芽分化以嫩叶愈伤组织为材料,以MS为基本培养基,L_(16)(4~2)正交试验设计,统计分析结果表明,6-BA、NAA对不定芽分化的影响达到极显著水平,最佳分化培养基培为:MS+6-BA0.50mg·L~(-1)+NAA0.10mg·L~(-1),分化率可达95%。
4.不定芽的增殖培养,采用L_(16)(4~2)正交试验设计,统计分析结果表明,培养基、NAA对继代增殖均达极显著水平,最佳增殖培养基为:1/2MS+NAA0.01mg·L~(-1),增殖系数达3.33。
5.生根培养阶段,大量元素、糖浓度、NAA均影响Sun Fan无菌苗生根,最适合的培养基为:1/2MS+NAA0.01mg·L~(-1)-0.02mg·L~(-1)+蔗糖30g·L~(-1)。
6.炼苗移栽需先闭瓶炼苗3-5d,然后开瓶炼苗3d,移栽到腐殖质:珍珠岩:细土=2:1:1的基质中,移栽成活率达75%以上。
采用嫩枝扦插法,研究了插条选择部位及长度、不同扦插基质、不同生长调节物质等对嫩枝插穗生根的促进效应。研究结果如下:
1.扦插基质对Sun Fan的扦插成活率具有极显著差异,细土:腐殖质:珍珠岩=1:2:1的混合基质,可以使扦插苗生根率达到80%以上。
2.插条部位对Sun Fan扦插生根率具有极显著差异,同一枝条但不同部位各段插条,从梢部、中部到基部,扦插苗的生根率依次降低。
3.插条长度对Sun Fan扦插的生根率具有极显著差异,插条长度为8cm时生根率最高。
4.植物生长调节剂对Sun Fan扦插的生根率具有极显著差异,NAA和IBA都能显著提高插条的生根率,达95%以上,但NAA处理扦插苗后期生长更好,用700mg·L~(-1)的NAA浸蘸插条基部10s是促进Sun Fan扦插生根的最佳激素条件。
Sun Fan(Scaevola albida),belonging to Scaevola,is one of perennial bulbous flowers,and famous with good adaptability and pealike colors.Since entering the flower market in 1989,the flower has become an important species in Australia market,and has been introduced to Europe and North America successfully.It has been widely used as a basket,and planted flowers,potted plants and cut flowers by consumers.However,there has no such flower in China,so it has high ornamental values.As the number of seedlings can not meet the needs of the market,this paper researched on the technology of Sun Fan tissue culture and cutting propagation to improve the quantity and quality of Sun Fan in an effective way.That can provide a theoretical and technical support for the industrialization of the production of nursery stock and excellent germplasm conservation.
Using tissue culture technique to explore the whole process from the aseptic system,the establishment of callus induction,adventitious bud differentiation and proliferation of culture, rooting culture to vitro culture.the conclusions as follows:
1.Twiges,leaves,and stems of Sun Fan are used to be explants,which are cultured by preculture and initial culture.Sterilization methods as follows:75%alcohol soaked 30s,0.1%HgCl_2 disinfection 3min which are cultured in MS medium supplemented with NAA(0.1mg·L~(-1)).The results showed that the highest rate of callus induction of leaves reaches 87.5%.Sun Fan leaves is the best tissue culture explants.
2.This paper researched on using twigs of Sunfan as explants which are cultured in MS medium,applying L_(16)(4~3)orthogonal experimental design to the experiment.Different hormones such as 6-BA、NAA、2,4-D are prominent to callus and the best combination is MS+6-BA0.50mg·L~(-1)+2,4-D0.20mg·L~(-1)+ NAA0.10mg·L~(-1) which the rate of induction is 100%.
3.Differentiation of adventitious buds to leaves callus material to the basic MS medium,L_(16) (4~2) orthogonal experimental design,it showed that different hormones of 6-BA,NAA are prominent to adventitious bud differentiation and the best combination is MS+6-BA0.50mg·L~(-1)+NAA0.10mg·L~(-1).
4.L_(16)(4~2) orthogonal experimental design is applied to culture adventitious buds.The result showed that the medium and NAA were prominent to proliferation,and the best medium for proliferation is:1/2MS+NAA0.01 mg·~L(-1).
5.During the step of rooting,a large number of elements,sugar concentration,NAA are all affect no vaccine of Sun Fan to take root,while,the most suitable medium for:1/2MS+ NAA0.01mg·L~(-1)-0.02mg·L~(-1) + Saccharose30g·L~(-1)。
6.3-5 hours are to be needed to cultivate twigs in bottles,then 3 hours are need to cultive twigs in air.The mixture of mixed fine soil:humus:perlite=1:2:1 could makes the rate of radication 75%.
The method of twigs cuttings is applied to study the suitable body,length of cuttings and growth regulators on rooting of twigs cuttings.The results are as follows:
1.The survival rate of Sun Fan cuttings is prominent for cutting medium.The mixture of mixed fine soil:humus:perlite=1:2:1 makes the rate of radication 80%.
2.The position of quickset is prominent to the rate of radication.The rooting rate is descending ordinals from top,middle to underside.
3.The length of quickset is prominent to the rate of radication,and the rate is the highest when the length is 8cm.
4.Plant regulator is prominent to the rate of radication.The effect of NAA is better than which of IBA in increasing the rate of redication.It is indicated that 700 mg·L~(-1) of NAA baptist cuttings dip the base of 10s for the plus hormone conditions is the best condition to increase the rate of redication.
采用组织培养技术手段,探讨了从无菌体系的建立、愈伤诱导、不定芽分化、增殖培养、生根培养到试管苗移栽的整个过程,经过研究分析,得出以下结论:
1.以Sun Fan嫩叶、老叶、茎段、茎节为外植体,消毒方式为:75%酒精浸泡30s,0.1%HgCl_2消毒3min,接种在MS基本培养基,附加0.1mg·L~(-1)NAA进行培养,试验结果表明,嫩叶愈伤组织诱导率最高,达87.5%;嫩叶为Sun Fan最佳组织培养外植体。
2.愈伤诱导以Sun Fan嫩叶为外植体,以MS为基本培养基,L_(16)(4~3)正交试验设计,统计分析结果表明,6-BA、2,4-D、NAA对Sun Fan嫩叶愈伤组织的诱导影响均达到极显著水平,最佳愈伤诱导培养基为:MS+6-BA0.50mg·L~(-1)+2,4-D0.20mg·L~(-1)+NAA0.10mg·L~(-1),诱导率高达100%。
3.不定芽分化以嫩叶愈伤组织为材料,以MS为基本培养基,L_(16)(4~2)正交试验设计,统计分析结果表明,6-BA、NAA对不定芽分化的影响达到极显著水平,最佳分化培养基培为:MS+6-BA0.50mg·L~(-1)+NAA0.10mg·L~(-1),分化率可达95%。
4.不定芽的增殖培养,采用L_(16)(4~2)正交试验设计,统计分析结果表明,培养基、NAA对继代增殖均达极显著水平,最佳增殖培养基为:1/2MS+NAA0.01mg·L~(-1),增殖系数达3.33。
5.生根培养阶段,大量元素、糖浓度、NAA均影响Sun Fan无菌苗生根,最适合的培养基为:1/2MS+NAA0.01mg·L~(-1)-0.02mg·L~(-1)+蔗糖30g·L~(-1)。
6.炼苗移栽需先闭瓶炼苗3-5d,然后开瓶炼苗3d,移栽到腐殖质:珍珠岩:细土=2:1:1的基质中,移栽成活率达75%以上。
采用嫩枝扦插法,研究了插条选择部位及长度、不同扦插基质、不同生长调节物质等对嫩枝插穗生根的促进效应。研究结果如下:
1.扦插基质对Sun Fan的扦插成活率具有极显著差异,细土:腐殖质:珍珠岩=1:2:1的混合基质,可以使扦插苗生根率达到80%以上。
2.插条部位对Sun Fan扦插生根率具有极显著差异,同一枝条但不同部位各段插条,从梢部、中部到基部,扦插苗的生根率依次降低。
3.插条长度对Sun Fan扦插的生根率具有极显著差异,插条长度为8cm时生根率最高。
4.植物生长调节剂对Sun Fan扦插的生根率具有极显著差异,NAA和IBA都能显著提高插条的生根率,达95%以上,但NAA处理扦插苗后期生长更好,用700mg·L~(-1)的NAA浸蘸插条基部10s是促进Sun Fan扦插生根的最佳激素条件。
Sun Fan(Scaevola albida),belonging to Scaevola,is one of perennial bulbous flowers,and famous with good adaptability and pealike colors.Since entering the flower market in 1989,the flower has become an important species in Australia market,and has been introduced to Europe and North America successfully.It has been widely used as a basket,and planted flowers,potted plants and cut flowers by consumers.However,there has no such flower in China,so it has high ornamental values.As the number of seedlings can not meet the needs of the market,this paper researched on the technology of Sun Fan tissue culture and cutting propagation to improve the quantity and quality of Sun Fan in an effective way.That can provide a theoretical and technical support for the industrialization of the production of nursery stock and excellent germplasm conservation.
Using tissue culture technique to explore the whole process from the aseptic system,the establishment of callus induction,adventitious bud differentiation and proliferation of culture, rooting culture to vitro culture.the conclusions as follows:
1.Twiges,leaves,and stems of Sun Fan are used to be explants,which are cultured by preculture and initial culture.Sterilization methods as follows:75%alcohol soaked 30s,0.1%HgCl_2 disinfection 3min which are cultured in MS medium supplemented with NAA(0.1mg·L~(-1)).The results showed that the highest rate of callus induction of leaves reaches 87.5%.Sun Fan leaves is the best tissue culture explants.
2.This paper researched on using twigs of Sunfan as explants which are cultured in MS medium,applying L_(16)(4~3)orthogonal experimental design to the experiment.Different hormones such as 6-BA、NAA、2,4-D are prominent to callus and the best combination is MS+6-BA0.50mg·L~(-1)+2,4-D0.20mg·L~(-1)+ NAA0.10mg·L~(-1) which the rate of induction is 100%.
3.Differentiation of adventitious buds to leaves callus material to the basic MS medium,L_(16) (4~2) orthogonal experimental design,it showed that different hormones of 6-BA,NAA are prominent to adventitious bud differentiation and the best combination is MS+6-BA0.50mg·L~(-1)+NAA0.10mg·L~(-1).
4.L_(16)(4~2) orthogonal experimental design is applied to culture adventitious buds.The result showed that the medium and NAA were prominent to proliferation,and the best medium for proliferation is:1/2MS+NAA0.01 mg·~L(-1).
5.During the step of rooting,a large number of elements,sugar concentration,NAA are all affect no vaccine of Sun Fan to take root,while,the most suitable medium for:1/2MS+ NAA0.01mg·L~(-1)-0.02mg·L~(-1) + Saccharose30g·L~(-1)。
6.3-5 hours are to be needed to cultivate twigs in bottles,then 3 hours are need to cultive twigs in air.The mixture of mixed fine soil:humus:perlite=1:2:1 could makes the rate of radication 75%.
The method of twigs cuttings is applied to study the suitable body,length of cuttings and growth regulators on rooting of twigs cuttings.The results are as follows:
1.The survival rate of Sun Fan cuttings is prominent for cutting medium.The mixture of mixed fine soil:humus:perlite=1:2:1 makes the rate of radication 80%.
2.The position of quickset is prominent to the rate of radication.The rooting rate is descending ordinals from top,middle to underside.
3.The length of quickset is prominent to the rate of radication,and the rate is the highest when the length is 8cm.
4.Plant regulator is prominent to the rate of radication.The effect of NAA is better than which of IBA in increasing the rate of redication.It is indicated that 700 mg·L~(-1) of NAA baptist cuttings dip the base of 10s for the plus hormone conditions is the best condition to increase the rate of redication.
引文
[1]Habedandt G.Kulturversuche Mit Isolierten Math-Natur-wiss Klasse Kais Akad Wiss Wien.1902,Pflanzenzellen[J].Sitzungsber,111:69-92.
[2]王家福.花卉组织培养与快繁技术[M].中国林业出版社,2006,172-176.
[3]李俊明.植物组织培养教程[M].中国农业大学出版社.2002,6:4-9.
[4]曹孜义,刘国民.实用植物组织培养技术教程[M].兰州:甘肃科学技术出版社,1996:1-8.
[5]潘瑞帜,懂愚得.植物生理学[M].北京高等教育出版社,1995:249-251.
[6]刘庆昌,吴国良.植物细胞组织培养[M].中国农业大学出版社,2005:35-37.
[7]DeberghP,AiykenChristieJ,CohenDetal.Reconsideration of the term viteification as used in micropropagation[J].Plant Cell Tiss Org Cult,1992,(30):135-140.
[8]王熊.地生兰个体发生途径研究[J].植物生理学报,1990.16(3):264.
[9]BrukhinVB,BatyginaTB.Embryo culture and somatic embryogenesis in culture of Paeonia anomala[J].Phytomorphology.1994,44(3-4) 151-157.
[10]韩碧文.植物组织培养研究的进展[M].北京:中国植物生理学史料汇编,1993:61.
[11]夏镇澳.植物组织培养与农业[J].植物生理学通讯,1995,(1):62.
[12]ChevreA M.In vitro vegetative multiplication of chestnut[J].J of Hort Sic.1983,58(1):23-29.
[13]周淘,张启翔.观赏花卉组织培养中外植体材料的选取[J].山东林业科技,2003,(1):43-44.
[14]王清连.植物组织培养[M].北京:中国农业出版社,2001.22-23.
[15]Wang Q C,Tang H R,QuanY et al.Phenolinduce browning and establishment of shoot-tip Explants of"Fuji" apple and "jinhua pear"culture in vitrol[J].J of Hort Sci.1994,69(5):833-839.
[16]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,2002.
[17]李合生等.植物生理生化原理和技术[M].北京:高等教育出版社,2000.
[18]詹亚光,陶静,杨传平,刘玉喜.白桦组培再生体系统的研究(Ⅱ)-影响因素及培养程序[J].东北林业大学学报,1998,26(6):1-5.
[19]李浚明.植物组织培养教程[M].北京:中国农业大学出版社,2002:65-67.
[20]刘青林,马炜,郑玉梅.花卉组织培养[M].北京:中国农业出版社.2003:41-45.
[21]张思文.关于我国古籍中插枝技术的初步研究[M].中国古代农业科技农业出版社,1980:54.
[22]王景章等.日本落叶松、杂种落叶松嫩枝全光喷雾扦插的研究[J].东北林业大学学报,1990,18(3):9-17.
[23]韩德元.植物生长调节剂--原理与应用[M].北京:科学技术出版社,1997.
[24]刘云龙.组织培养中植物生长调节剂的调控[M].农业与技术,1996.
[25]裎广有名优花卉组织培养技术[M].北京:学技术文献出版,2001:35.
[26]童朝阳.菊花组织培养和快速繁殖[J].芜湖职业技术学院学报,2001,3(1):28-29.
[27]张晓军,任如意,赵金良月季组织离体培养及快速繁殖研究[J].牡丹江师范学院学报,2001(2):14-15.
[28]李艳敏,罗晓芳.牡丹离体培养与快速繁殖研究进展[J].西南林学院学报,2004,24(1):70-73.
[29]赵慧.云南山茶花快速繁殖技术[J].中国土特产,1997,1:16-17.
[30]刘燕.园林花卉学[M].北京:中国林业出版社,2003,3.
[31]王晶英主编.植物生理生化实验技术与原理[M].东北林业大学出版社,2003.
[32]刘鹏,刘金,等.菊花的组织培养、脱毒与快繁技术研究[J].内蒙古民族大学学报,2005,20(4):410-413.
[33]吴延军,周慧民.香石竹茎尖脱毒技术[J].西南园艺,2003,31(1):20.
[34]袁学军,郑家焕.水仙脱毒快繁[J].植物杂志,1999,2:28.
[35]满贵武,孙冬荣,等.大丽花脱毒快繁技术[J].北方园艺,2002,4:63.
[36]朱根发,王怀宇.文心兰脱毒快繁技术研究[J].广东农业科学.1997,4:27-28.
[37]程治英等.植物遗传多样性离体保存的研究[M].四川大学出版社,1996:55-57.
[38]王洁琼,张启翔.百合产业化发展[J].中国观赏园艺研究进展,2005.8:565-568.
[39]杨文钮等.植物化控[M].四川科学技术出版社,1997.
[40]季孔庶.杂交鹅掌揪的无性繁殖[J].南京林业大学学报(白然科学版),2005,29(1):83-87.
[41]刘继梅,鄢波等.不同花色矮牵牛细胞色素b5蛋白的cDNA克隆及序列分析[J].云南植物研究,2002,24(2):215-221.
[42]陈慕英,于喜水.甜菊甙类的研究[J].中医药信息,2001,18(3):23-24.
[43]祖元刚,罗猛,等.长春花生物碱成分及其药理作用研究进展[J].天然产物研究与开发,2006,18(2):325-329.
[44]李俊,牙启康等.人工栽培南方红豆杉紫杉烷成分的研究[J].广西师范大学学报(自然科学版),2006,24(1):72-74.
[45]Hartman H T.Effect of season of collecting idolebutyric acid and pre-planting storage treatment on rooting of Marianna plum.peach and Quinse hardwood cutting pro Amor Soc Hort ser.1958,(71):57-66.
[46]毕洪昱,张学科.花木扦插培育的研究[J].东北林业大学学报,1989,17(4):97-102.
[47]戴台金,施新程,金申燕.不同基质对美国侧柏扦插生根的影响[J].林业科技,2004,29(1):53-54.
[48]王关林,苏冬霞.代谢调节剂对嫩枝扦插繁殖成括率的影响及其机理[J],园艺学报,2006,33(2):395-398.
[49]王军辉,张建国,张守攻,等.青海云杉硬枝扦插的激素、年龄和位置研究效应研究[J].西北农林科技大学学报(自然科学版),2006,34(7):65-71.
[50]郑均宝,刘玉军,等.几种木本植物插穗生根与内源IAA,ABA的关系[J].植物生理学报,1991,17(3):313-316.
[51]段新玲,任东岁,段黄金.影响蓝花大叶醉鱼草嫩枝扦插成活因素的研究[J].西北林学院学报,1999,14(4):85-88.
[52]王小明,马耀祖,李健.几种处理方法对四翅滨藜嫩枝扦插成活率影响的研究[J].西部林业科学,2006,35(4):115-117.
[53]Nyirenda H E.Vegetative Propagation of tea using immature(Apical shoots)and overmature (Brownstem)cuttings[J].TRF QNL,1995,(119):12-13.
[54]吕文.难生根树种嫩枝扦插技术及生理机理的研究[J].防护林科技,1993,16(3):14-20.
[55]余发新,刘腾云,朱祺,高柱,江洪如,SE碧琴,周华.杂种马褂木扦插繁殖技术研究-Ⅱ插穗粗细及环境条件与生根的关系[J].江西科学,2006,(01).
[56]杨秋,唐岱,苏腾伟,等.基质及生长调节剂对切花小菊扦插生根的影响[J].安徽农业科学,2007,35(2):416-418.
[57]顾永华,杨军,何云,等.秤锤树扦插繁殖技术[J].林业科技开发,2007,21(1):34-36.
[58]韩国勇.南方红豆杉嫩枝扦插育苗技术[J].林业科技开发,2006,20(5):87-88.
[59]程水源,罗晓.果树扦插繁殖研究进展[J].湖北农学院学报,1992,12(2):57-62.
[60]孙时轩.林木育苗技术[M].北京:金盾出版社,2002,3.
[61]Haissig B E.Influence of auxthin and auxin synergists on adventious root primordium in initiation and development[J].New Zealand Journal Forestry Science,1974,4(2):311-323.
[62]李继华.扦插的原理与应用[M].上海:上海科学技术出版社,1987.
[63]武维华.植物生理学[M].北京:科学出版社,2003.
[64]齐秀东,郭守华,于凤鸣,等.NAA和IBA对金露梅扦插生根的作用[J].江苏农业科学,2007,(1):103-105.
[65]王关林.NAA、IBA对樱桃砧木插条的生理、生化代谢和生根的影响[J].园艺学报,2005,32(4):691-694.
[66]H.T.哈特曼,D.E.凯斯特著,郑开文译.植物繁殖原理和技术[M].中国林业出版,1981.
[67]Schumarm I.Control of flowering in Scaevola[J].Gaertnerboerse und Gartenwelt.1991(91):1709-1714.
[68]Steffen,K.A hanging basket plant with future-Scaevola aemula[J].Gaertnermeister,1989(92):944-945.
[69]Soo-Hyung Kim,Paul R.Fisher,J.Heinrich Lieth,Analysis and modeling of gas exchange processes in Scaevola aemula[J].Scientia Horticulturae,2007.(114):170-176.
[70]Zhang,D.,Moran,R.E.,Stack,L.B.Effect of phosphorus fertilization on growth and flowering of Seaevola aemula R.Br.[J].'NewWonder'HortScience,2004(39):1728-1731.
[71]Mandal AKA,Gupta SD,Chatterji AK.Factors affecting somatic embryogenesis from cotyledonary explants of sunflower[J].Biol Plant,2001(44):503-507.
[72]Swoboda I,Bhalla P.L.RAPD analysis of genetic variation in the Australian fan flower,Scaevola[J].Genome,1997(40):600-606.
[73]Bhalla P.L.,Xu H.Plant regeneration from callus of Australian fan flower,Scaevola[J].Plant Physiol,1999(154):374-378.
[74]Prem L.Bhalla,Katherine Sweeney.Direct in vitro regeneration of the Australian fan flower.Scaevola aemula R.Br[J].Scientia Horticulturae,1999(79):6-74.
[75]Wang Y.H.,Bhalla P.L.Somatic embryogenesis from leaf explants of Australian fan flower.Scaevola aemula R.Br[J].Plant Cell,2004(22):408-414.
[76]张兴翠,梁国鲁.互叶白干层组织快繁及多倍体诱导[J].西南农业大学学报,2000,22(6):507-509.
[77]刘权等.果树试验设计及统计[M].中国农业出版社.1997,148-153.
[78]康冰,于福科,张广军,等.应用正交试验筛选玫瑰茎段增殖培养基[J].西北植物学报,2003,23(4):653-655.
[79]王兴仁,张录达,王华方.正交试验设置重复的必要性和统计分析方法[J].土壤通报,2000,31(3):135-139.
[80]潘远智.一品红组织培养技术体系研究[J].四川农业大学学报,2001,6.
[81]张施君,王凤兰,周厚高等.火百合的组织培养及快速繁殖[J].江苏农业科学,2004,4:72-73.
[82]刘庆昌,吴国良.植物细胞组织培养[M].中国农业大学出版社,2005:35-37.
[83]Masaru Nakano,Shigefumi Tanaka,Miki Oota,Eisuke Ookawa,Sakae Suzuki&Hiroyuki Saito.Regeneration of diploid and tetraploid plants from callus-derived protoplasts of Agapanthus praecox spp.orientalis(Leighton) Leighton.Plant Cell,Tissue and Organ Culture,2003:63-69.
[84]Zenk,M.H.EL-Shasi,H.Arense-de.Plant tissue culture and its Biotechnological Application.Sprinser-Vedag.Bedin,1977,P.27-43.
[85]Shimada T.Plant regeneration from callus induced from wheat embryo Jap.J.Genet,1978,53:374-381.
[86]段安安.河北杨优树根萌条诱导及嫩枝扦插的研究[J].西北林学院学报,1988,3(1):9-16.
[2]王家福.花卉组织培养与快繁技术[M].中国林业出版社,2006,172-176.
[3]李俊明.植物组织培养教程[M].中国农业大学出版社.2002,6:4-9.
[4]曹孜义,刘国民.实用植物组织培养技术教程[M].兰州:甘肃科学技术出版社,1996:1-8.
[5]潘瑞帜,懂愚得.植物生理学[M].北京高等教育出版社,1995:249-251.
[6]刘庆昌,吴国良.植物细胞组织培养[M].中国农业大学出版社,2005:35-37.
[7]DeberghP,AiykenChristieJ,CohenDetal.Reconsideration of the term viteification as used in micropropagation[J].Plant Cell Tiss Org Cult,1992,(30):135-140.
[8]王熊.地生兰个体发生途径研究[J].植物生理学报,1990.16(3):264.
[9]BrukhinVB,BatyginaTB.Embryo culture and somatic embryogenesis in culture of Paeonia anomala[J].Phytomorphology.1994,44(3-4) 151-157.
[10]韩碧文.植物组织培养研究的进展[M].北京:中国植物生理学史料汇编,1993:61.
[11]夏镇澳.植物组织培养与农业[J].植物生理学通讯,1995,(1):62.
[12]ChevreA M.In vitro vegetative multiplication of chestnut[J].J of Hort Sic.1983,58(1):23-29.
[13]周淘,张启翔.观赏花卉组织培养中外植体材料的选取[J].山东林业科技,2003,(1):43-44.
[14]王清连.植物组织培养[M].北京:中国农业出版社,2001.22-23.
[15]Wang Q C,Tang H R,QuanY et al.Phenolinduce browning and establishment of shoot-tip Explants of"Fuji" apple and "jinhua pear"culture in vitrol[J].J of Hort Sci.1994,69(5):833-839.
[16]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,2002.
[17]李合生等.植物生理生化原理和技术[M].北京:高等教育出版社,2000.
[18]詹亚光,陶静,杨传平,刘玉喜.白桦组培再生体系统的研究(Ⅱ)-影响因素及培养程序[J].东北林业大学学报,1998,26(6):1-5.
[19]李浚明.植物组织培养教程[M].北京:中国农业大学出版社,2002:65-67.
[20]刘青林,马炜,郑玉梅.花卉组织培养[M].北京:中国农业出版社.2003:41-45.
[21]张思文.关于我国古籍中插枝技术的初步研究[M].中国古代农业科技农业出版社,1980:54.
[22]王景章等.日本落叶松、杂种落叶松嫩枝全光喷雾扦插的研究[J].东北林业大学学报,1990,18(3):9-17.
[23]韩德元.植物生长调节剂--原理与应用[M].北京:科学技术出版社,1997.
[24]刘云龙.组织培养中植物生长调节剂的调控[M].农业与技术,1996.
[25]裎广有名优花卉组织培养技术[M].北京:学技术文献出版,2001:35.
[26]童朝阳.菊花组织培养和快速繁殖[J].芜湖职业技术学院学报,2001,3(1):28-29.
[27]张晓军,任如意,赵金良月季组织离体培养及快速繁殖研究[J].牡丹江师范学院学报,2001(2):14-15.
[28]李艳敏,罗晓芳.牡丹离体培养与快速繁殖研究进展[J].西南林学院学报,2004,24(1):70-73.
[29]赵慧.云南山茶花快速繁殖技术[J].中国土特产,1997,1:16-17.
[30]刘燕.园林花卉学[M].北京:中国林业出版社,2003,3.
[31]王晶英主编.植物生理生化实验技术与原理[M].东北林业大学出版社,2003.
[32]刘鹏,刘金,等.菊花的组织培养、脱毒与快繁技术研究[J].内蒙古民族大学学报,2005,20(4):410-413.
[33]吴延军,周慧民.香石竹茎尖脱毒技术[J].西南园艺,2003,31(1):20.
[34]袁学军,郑家焕.水仙脱毒快繁[J].植物杂志,1999,2:28.
[35]满贵武,孙冬荣,等.大丽花脱毒快繁技术[J].北方园艺,2002,4:63.
[36]朱根发,王怀宇.文心兰脱毒快繁技术研究[J].广东农业科学.1997,4:27-28.
[37]程治英等.植物遗传多样性离体保存的研究[M].四川大学出版社,1996:55-57.
[38]王洁琼,张启翔.百合产业化发展[J].中国观赏园艺研究进展,2005.8:565-568.
[39]杨文钮等.植物化控[M].四川科学技术出版社,1997.
[40]季孔庶.杂交鹅掌揪的无性繁殖[J].南京林业大学学报(白然科学版),2005,29(1):83-87.
[41]刘继梅,鄢波等.不同花色矮牵牛细胞色素b5蛋白的cDNA克隆及序列分析[J].云南植物研究,2002,24(2):215-221.
[42]陈慕英,于喜水.甜菊甙类的研究[J].中医药信息,2001,18(3):23-24.
[43]祖元刚,罗猛,等.长春花生物碱成分及其药理作用研究进展[J].天然产物研究与开发,2006,18(2):325-329.
[44]李俊,牙启康等.人工栽培南方红豆杉紫杉烷成分的研究[J].广西师范大学学报(自然科学版),2006,24(1):72-74.
[45]Hartman H T.Effect of season of collecting idolebutyric acid and pre-planting storage treatment on rooting of Marianna plum.peach and Quinse hardwood cutting pro Amor Soc Hort ser.1958,(71):57-66.
[46]毕洪昱,张学科.花木扦插培育的研究[J].东北林业大学学报,1989,17(4):97-102.
[47]戴台金,施新程,金申燕.不同基质对美国侧柏扦插生根的影响[J].林业科技,2004,29(1):53-54.
[48]王关林,苏冬霞.代谢调节剂对嫩枝扦插繁殖成括率的影响及其机理[J],园艺学报,2006,33(2):395-398.
[49]王军辉,张建国,张守攻,等.青海云杉硬枝扦插的激素、年龄和位置研究效应研究[J].西北农林科技大学学报(自然科学版),2006,34(7):65-71.
[50]郑均宝,刘玉军,等.几种木本植物插穗生根与内源IAA,ABA的关系[J].植物生理学报,1991,17(3):313-316.
[51]段新玲,任东岁,段黄金.影响蓝花大叶醉鱼草嫩枝扦插成活因素的研究[J].西北林学院学报,1999,14(4):85-88.
[52]王小明,马耀祖,李健.几种处理方法对四翅滨藜嫩枝扦插成活率影响的研究[J].西部林业科学,2006,35(4):115-117.
[53]Nyirenda H E.Vegetative Propagation of tea using immature(Apical shoots)and overmature (Brownstem)cuttings[J].TRF QNL,1995,(119):12-13.
[54]吕文.难生根树种嫩枝扦插技术及生理机理的研究[J].防护林科技,1993,16(3):14-20.
[55]余发新,刘腾云,朱祺,高柱,江洪如,SE碧琴,周华.杂种马褂木扦插繁殖技术研究-Ⅱ插穗粗细及环境条件与生根的关系[J].江西科学,2006,(01).
[56]杨秋,唐岱,苏腾伟,等.基质及生长调节剂对切花小菊扦插生根的影响[J].安徽农业科学,2007,35(2):416-418.
[57]顾永华,杨军,何云,等.秤锤树扦插繁殖技术[J].林业科技开发,2007,21(1):34-36.
[58]韩国勇.南方红豆杉嫩枝扦插育苗技术[J].林业科技开发,2006,20(5):87-88.
[59]程水源,罗晓.果树扦插繁殖研究进展[J].湖北农学院学报,1992,12(2):57-62.
[60]孙时轩.林木育苗技术[M].北京:金盾出版社,2002,3.
[61]Haissig B E.Influence of auxthin and auxin synergists on adventious root primordium in initiation and development[J].New Zealand Journal Forestry Science,1974,4(2):311-323.
[62]李继华.扦插的原理与应用[M].上海:上海科学技术出版社,1987.
[63]武维华.植物生理学[M].北京:科学出版社,2003.
[64]齐秀东,郭守华,于凤鸣,等.NAA和IBA对金露梅扦插生根的作用[J].江苏农业科学,2007,(1):103-105.
[65]王关林.NAA、IBA对樱桃砧木插条的生理、生化代谢和生根的影响[J].园艺学报,2005,32(4):691-694.
[66]H.T.哈特曼,D.E.凯斯特著,郑开文译.植物繁殖原理和技术[M].中国林业出版,1981.
[67]Schumarm I.Control of flowering in Scaevola[J].Gaertnerboerse und Gartenwelt.1991(91):1709-1714.
[68]Steffen,K.A hanging basket plant with future-Scaevola aemula[J].Gaertnermeister,1989(92):944-945.
[69]Soo-Hyung Kim,Paul R.Fisher,J.Heinrich Lieth,Analysis and modeling of gas exchange processes in Scaevola aemula[J].Scientia Horticulturae,2007.(114):170-176.
[70]Zhang,D.,Moran,R.E.,Stack,L.B.Effect of phosphorus fertilization on growth and flowering of Seaevola aemula R.Br.[J].'NewWonder'HortScience,2004(39):1728-1731.
[71]Mandal AKA,Gupta SD,Chatterji AK.Factors affecting somatic embryogenesis from cotyledonary explants of sunflower[J].Biol Plant,2001(44):503-507.
[72]Swoboda I,Bhalla P.L.RAPD analysis of genetic variation in the Australian fan flower,Scaevola[J].Genome,1997(40):600-606.
[73]Bhalla P.L.,Xu H.Plant regeneration from callus of Australian fan flower,Scaevola[J].Plant Physiol,1999(154):374-378.
[74]Prem L.Bhalla,Katherine Sweeney.Direct in vitro regeneration of the Australian fan flower.Scaevola aemula R.Br[J].Scientia Horticulturae,1999(79):6-74.
[75]Wang Y.H.,Bhalla P.L.Somatic embryogenesis from leaf explants of Australian fan flower.Scaevola aemula R.Br[J].Plant Cell,2004(22):408-414.
[76]张兴翠,梁国鲁.互叶白干层组织快繁及多倍体诱导[J].西南农业大学学报,2000,22(6):507-509.
[77]刘权等.果树试验设计及统计[M].中国农业出版社.1997,148-153.
[78]康冰,于福科,张广军,等.应用正交试验筛选玫瑰茎段增殖培养基[J].西北植物学报,2003,23(4):653-655.
[79]王兴仁,张录达,王华方.正交试验设置重复的必要性和统计分析方法[J].土壤通报,2000,31(3):135-139.
[80]潘远智.一品红组织培养技术体系研究[J].四川农业大学学报,2001,6.
[81]张施君,王凤兰,周厚高等.火百合的组织培养及快速繁殖[J].江苏农业科学,2004,4:72-73.
[82]刘庆昌,吴国良.植物细胞组织培养[M].中国农业大学出版社,2005:35-37.
[83]Masaru Nakano,Shigefumi Tanaka,Miki Oota,Eisuke Ookawa,Sakae Suzuki&Hiroyuki Saito.Regeneration of diploid and tetraploid plants from callus-derived protoplasts of Agapanthus praecox spp.orientalis(Leighton) Leighton.Plant Cell,Tissue and Organ Culture,2003:63-69.
[84]Zenk,M.H.EL-Shasi,H.Arense-de.Plant tissue culture and its Biotechnological Application.Sprinser-Vedag.Bedin,1977,P.27-43.
[85]Shimada T.Plant regeneration from callus induced from wheat embryo Jap.J.Genet,1978,53:374-381.
[86]段安安.河北杨优树根萌条诱导及嫩枝扦插的研究[J].西北林学院学报,1988,3(1):9-16.