大蒜微繁的一种新技术体系的构建
|
摘要
大蒜是一种栽培历史悠久的世界性蔬菜，主要靠鳞茎进行无性繁殖。大蒜的无性繁殖方式给大蒜的生产带来两个问题：一无性繁殖使大蒜病毒病积累，且广泛传播；二繁殖系数低。应用茎尖分生组织培养获得脱毒苗，是脱除大蒜病毒的行之有效的方法。大蒜脱毒苗培育技术现已基本成熟，目前许多品种都已获得脱毒苗。但大蒜离体培养繁殖系数过低，脱毒种苗生产成本相当昂贵，成为大蒜脱毒苗进入生产实用化的瓶颈。
一个蒜瓣可发出几十条根，数量多，取材方便，使笔者受到启发，以大蒜蒜瓣发出的根的根尖为外植体，对影响大蒜根尖直接分化不定芽的各种因素进行了详细的探讨，并进一步研究了根尖分化的不定芽的生长与增殖情况，旨在建立一套取材方便、操作简单、繁殖系数高的大蒜微繁新技术体系，以突破上述瓶颈，推动大蒜脱毒苗生产进入实用化、产业化。并对蔗糖及乙烯、PP_(333)两种生长抑制剂对鳞茎的形成与膨大效果进行了探讨，旨在增加试管鳞茎重量，促使试管鳞茎分瓣，进一步提高大蒜微繁系数。同时对试管鳞茎移栽后零代与子一代的情况作了调查，以验证根尖微繁体系的实用性。
1．大蒜根尖培养诱导不定芽
通过研究不同大蒜品种、蒜瓣生根培养时间、根尖长度及不同培养条件(温度，光照时间，培养基，糖的种类与浓度，pH值，激素配比等)对根尖直接分化不定芽的影响，表明本论文所研究的四个大蒜品种(徐州白蒜，太仓白蒜，二水早，金乡蒜)根尖分化不定芽的频率及不定芽数目存在品种间差异；在蒜瓣生根培养时间为4～8天时，切取长度为2～3mm的蒜瓣根尖进行培养，根尖分化不定芽的频率最高。由不同外植体诱导生成的试管苗再诱导生出的根的根尖均能直接分化不定芽，说明大蒜可通过根尖进行循环增殖，但蒜瓣发出的根的根尖分化不定芽的频率较高。大蒜根尖培养分化不定芽需要较高的温度，在15℃、20℃下培养时，根尖全部生长成为白色的根组织，没有不定芽的发生；在28℃、32℃下培养，根尖不定芽分化频率及不定芽数显著提高。长的光照时间能促进根尖分化不定芽，大幅度提高不定芽数目。不同基本培养基、糖的种类与浓度、pH值、激素配比等对根尖分化不定芽的影响的研究表明，根尖培养最适宜的基本培养基为MS盐+B+5有机和MS培养基，蔗糖20～50g／L或白糖30g／L，BA2.0～5.0mg／L，NAA1.0mg／L，pH5.8～6.2。
2．大蒜根尖分化的不定芽的继代培养
以大蒜根尖培养诱导分化的不定芽为试验材料进行继代培养，以调查研究根尖在
不同的根尖启动培养基上分化的不定芽的生长与增殖情况，不同基掷养基、不同激
素浓度组合对大蒜根尖分化的不定芽的生长与增殖的影响，结果表明，根尖在含
BAI omg／L，NAAlmg／L的起动培养基上分化的不定芽在继代培养时生成的试管苗最多，
根尖在含M吧／L，NAAO．ling／L起动培养基上分化的不定芽在继代培养时的长势最强；
不定芽生长的最适培养基为MS盐巾；有机或B；刊．oms／LBA叩．05mg儿NAA，不定芽增殖
的最适培养基为MS盐＊；有机或B；＋0．sing／LBA。
3．大蒜试管鳞茎的培育与田间表现
以 B；为基本培养基，研究蔗糖浓度、乙烯、PP；；；对大蒜试管鳞茎形成与膨大的影
响，表明蔗糖浓度提高到9 og／L能显著提高鳞茎形成率与重量；加入乙烯、PP；；；对鳞
茎的形成与生长均有显著促进作用；在BS叫0g／L蔗糖八．0 mg／LPP；；；的培养基上，鳞
茎生长效果最佳，可达31hg；在BS叫0；／L蔗糖d．0 m；／LPP；；；的培养基上，大蒜试
管鳞茎能分瓣，分瓣率达33．1％。试管鳞茎的大小与幼苗的萌发、生长及鳞茎形咸的
大小相关，3 0 omg以上的鳞茎萌发率高，生长健壮，能形成较重的鳞茎。试管鳞茎移
栽后，零代生长正常，但植株矮小，形成的鳞茎小，多数为独头蒜，少数分瓣，但瓣
数少；子一代生长正常，植株的叶数、株高、最大叶宽与零代相比，均显著增大，形
成的鳞茎也能正常分瓣；鳞茎瓣数、重量与零代相比，显著增大，但与对照相比要小，
可望在子二代达到与对照无差异或超过对照。
Garlic (Allium sativum L.) is a worldwide vegetable and had been cultivated since ancient times. Due to its sexual sterility, garlic is propagated vegetatively through cloves or bulbs. Two main problems associated with the vegetative propagation of garlic are (1) perpetuation and wide spread of viral infection and (2) low propagation rate.
Shoot-tip culture is proved to be an effective method for producing virus-free garlic seedlings. However, because of low micropropagation rate, virus-free plantlets are still very expensive which limits their practical use.
This study focuses on direct shoot regeneration from garlic root tips and shoot proliferation for speeding up multipropagation of virus-free garlic.
1. Induction of the adventitious buds from root tips in garlic
Using MS as basic medium, the effect of different varieties, rooting time, root-tip length and culture condition (temperature, photoperiod, basic media, sugar sources and concentration, pH value and different hormone concentration and combinations) on direct adventitious shoot formation from garlic root tips was studied. The results showed that 2~3mm root tips from the cloves rooting for 4-8 days have a high frequency bud initiation. The induction rate and the number of adventitious buds varied with varieties. Root tips from different rooted shoots induced from different
explants all can directly regenerate adventitious buds and virus-free garlic can be cycly multiplicated through root-tip culture. The differentiation rate of buds of root tips inducted from plantlets from stem tips and root tips and adventitious shoot number was low than that from garlic cloves. Root tips had no bud regeneration when they were cultured at 15C or 20 C and the induction rate and the number of buds increased with the rise of temperature when they were cultured above 28 C. Continuous light had a positive effect on adventitious bud formation and number. The optimal medium for root tip culture was MS salt+B5 organic medium or MS containing sucrose 20~50g/L, BA 2.0~5.0mg/L, NAA1.0mg/L,at pH5.8~6.2.
2. Subculture of adventitious buds from root tips in garlic
The growth and proliferation of adventitious buds from different initiation media were investigated. The results indicted that there were more shoots per explant on the initiation medium containing lOmg/LBA, Img/LNAA and adventitious buds from the initiation medium containing 2.0mg/LBA, 0.Img/LNAA grow vigorously. The optimal medium for adventitious bud growth was MS salt+B, organic (or 65) +1.0mg/LBA+0.05mg/LNAA and The optimal medium for adventitious bud proliferation was and MS salt+Bs organic (or 85) +0.5mg/LBA.
3. Bulblet formation and swelling of Garlic in vitro and performance in field
85 as basic medium, the study on effect of sucrose concentration ,ethephon and PPsss on bulblet induction and swelling of Garlic in vitro demonstrated that bulblet formation and
swelling were significantly enhanced by increasing sucrose to 90g/L, Ethephon and had positive effects on bulblet fomation and swelling, on the 85 medium supplemented with sucrose 90g/L, PP333 1.0mg/L, bulblet swelled best, fresh weight can reach 312mg, and on the medium containing 90g/L sucrose and
2.0mg/LPPs33, 33.1% bulblets tillered. Bulblet fresh weight was related with seedlings germination, growth, and bulb formation. Bulblet above 300mg had higher germination rate, seedlings grew strong and produced heavier bulb. Bulblets were transplanted into pots, and zero generation grew normally, but
plants were short and produced small bulbs with few clovs. First generation grew normally, and plant leaf number, height, leaf width significantly increased compared with zero generation. Ffirst generation can form much heavier bulb than zero generation with more clovs, but still lighter compared with
control. It is possible that second generation has no difference with control or surpass control.
一个蒜瓣可发出几十条根，数量多，取材方便，使笔者受到启发，以大蒜蒜瓣发出的根的根尖为外植体，对影响大蒜根尖直接分化不定芽的各种因素进行了详细的探讨，并进一步研究了根尖分化的不定芽的生长与增殖情况，旨在建立一套取材方便、操作简单、繁殖系数高的大蒜微繁新技术体系，以突破上述瓶颈，推动大蒜脱毒苗生产进入实用化、产业化。并对蔗糖及乙烯、PP_(333)两种生长抑制剂对鳞茎的形成与膨大效果进行了探讨，旨在增加试管鳞茎重量，促使试管鳞茎分瓣，进一步提高大蒜微繁系数。同时对试管鳞茎移栽后零代与子一代的情况作了调查，以验证根尖微繁体系的实用性。
1．大蒜根尖培养诱导不定芽
通过研究不同大蒜品种、蒜瓣生根培养时间、根尖长度及不同培养条件(温度，光照时间，培养基，糖的种类与浓度，pH值，激素配比等)对根尖直接分化不定芽的影响，表明本论文所研究的四个大蒜品种(徐州白蒜，太仓白蒜，二水早，金乡蒜)根尖分化不定芽的频率及不定芽数目存在品种间差异；在蒜瓣生根培养时间为4～8天时，切取长度为2～3mm的蒜瓣根尖进行培养，根尖分化不定芽的频率最高。由不同外植体诱导生成的试管苗再诱导生出的根的根尖均能直接分化不定芽，说明大蒜可通过根尖进行循环增殖，但蒜瓣发出的根的根尖分化不定芽的频率较高。大蒜根尖培养分化不定芽需要较高的温度，在15℃、20℃下培养时，根尖全部生长成为白色的根组织，没有不定芽的发生；在28℃、32℃下培养，根尖不定芽分化频率及不定芽数显著提高。长的光照时间能促进根尖分化不定芽，大幅度提高不定芽数目。不同基本培养基、糖的种类与浓度、pH值、激素配比等对根尖分化不定芽的影响的研究表明，根尖培养最适宜的基本培养基为MS盐+B+5有机和MS培养基，蔗糖20～50g／L或白糖30g／L，BA2.0～5.0mg／L，NAA1.0mg／L，pH5.8～6.2。
2．大蒜根尖分化的不定芽的继代培养
以大蒜根尖培养诱导分化的不定芽为试验材料进行继代培养，以调查研究根尖在
不同的根尖启动培养基上分化的不定芽的生长与增殖情况，不同基掷养基、不同激
素浓度组合对大蒜根尖分化的不定芽的生长与增殖的影响，结果表明，根尖在含
BAI omg／L，NAAlmg／L的起动培养基上分化的不定芽在继代培养时生成的试管苗最多，
根尖在含M吧／L，NAAO．ling／L起动培养基上分化的不定芽在继代培养时的长势最强；
不定芽生长的最适培养基为MS盐巾；有机或B；刊．oms／LBA叩．05mg儿NAA，不定芽增殖
的最适培养基为MS盐＊；有机或B；＋0．sing／LBA。
3．大蒜试管鳞茎的培育与田间表现
以 B；为基本培养基，研究蔗糖浓度、乙烯、PP；；；对大蒜试管鳞茎形成与膨大的影
响，表明蔗糖浓度提高到9 og／L能显著提高鳞茎形成率与重量；加入乙烯、PP；；；对鳞
茎的形成与生长均有显著促进作用；在BS叫0g／L蔗糖八．0 mg／LPP；；；的培养基上，鳞
茎生长效果最佳，可达31hg；在BS叫0；／L蔗糖d．0 m；／LPP；；；的培养基上，大蒜试
管鳞茎能分瓣，分瓣率达33．1％。试管鳞茎的大小与幼苗的萌发、生长及鳞茎形咸的
大小相关，3 0 omg以上的鳞茎萌发率高，生长健壮，能形成较重的鳞茎。试管鳞茎移
栽后，零代生长正常，但植株矮小，形成的鳞茎小，多数为独头蒜，少数分瓣，但瓣
数少；子一代生长正常，植株的叶数、株高、最大叶宽与零代相比，均显著增大，形
成的鳞茎也能正常分瓣；鳞茎瓣数、重量与零代相比，显著增大，但与对照相比要小，
可望在子二代达到与对照无差异或超过对照。
Garlic (Allium sativum L.) is a worldwide vegetable and had been cultivated since ancient times. Due to its sexual sterility, garlic is propagated vegetatively through cloves or bulbs. Two main problems associated with the vegetative propagation of garlic are (1) perpetuation and wide spread of viral infection and (2) low propagation rate.
Shoot-tip culture is proved to be an effective method for producing virus-free garlic seedlings. However, because of low micropropagation rate, virus-free plantlets are still very expensive which limits their practical use.
This study focuses on direct shoot regeneration from garlic root tips and shoot proliferation for speeding up multipropagation of virus-free garlic.
1. Induction of the adventitious buds from root tips in garlic
Using MS as basic medium, the effect of different varieties, rooting time, root-tip length and culture condition (temperature, photoperiod, basic media, sugar sources and concentration, pH value and different hormone concentration and combinations) on direct adventitious shoot formation from garlic root tips was studied. The results showed that 2~3mm root tips from the cloves rooting for 4-8 days have a high frequency bud initiation. The induction rate and the number of adventitious buds varied with varieties. Root tips from different rooted shoots induced from different
explants all can directly regenerate adventitious buds and virus-free garlic can be cycly multiplicated through root-tip culture. The differentiation rate of buds of root tips inducted from plantlets from stem tips and root tips and adventitious shoot number was low than that from garlic cloves. Root tips had no bud regeneration when they were cultured at 15C or 20 C and the induction rate and the number of buds increased with the rise of temperature when they were cultured above 28 C. Continuous light had a positive effect on adventitious bud formation and number. The optimal medium for root tip culture was MS salt+B5 organic medium or MS containing sucrose 20~50g/L, BA 2.0~5.0mg/L, NAA1.0mg/L,at pH5.8~6.2.
2. Subculture of adventitious buds from root tips in garlic
The growth and proliferation of adventitious buds from different initiation media were investigated. The results indicted that there were more shoots per explant on the initiation medium containing lOmg/LBA, Img/LNAA and adventitious buds from the initiation medium containing 2.0mg/LBA, 0.Img/LNAA grow vigorously. The optimal medium for adventitious bud growth was MS salt+B, organic (or 65) +1.0mg/LBA+0.05mg/LNAA and The optimal medium for adventitious bud proliferation was and MS salt+Bs organic (or 85) +0.5mg/LBA.
3. Bulblet formation and swelling of Garlic in vitro and performance in field
85 as basic medium, the study on effect of sucrose concentration ,ethephon and PPsss on bulblet induction and swelling of Garlic in vitro demonstrated that bulblet formation and
swelling were significantly enhanced by increasing sucrose to 90g/L, Ethephon and had positive effects on bulblet fomation and swelling, on the 85 medium supplemented with sucrose 90g/L, PP333 1.0mg/L, bulblet swelled best, fresh weight can reach 312mg, and on the medium containing 90g/L sucrose and
2.0mg/LPPs33, 33.1% bulblets tillered. Bulblet fresh weight was related with seedlings germination, growth, and bulb formation. Bulblet above 300mg had higher germination rate, seedlings grew strong and produced heavier bulb. Bulblets were transplanted into pots, and zero generation grew normally, but
plants were short and produced small bulbs with few clovs. First generation grew normally, and plant leaf number, height, leaf width significantly increased compared with zero generation. Ffirst generation can form much heavier bulb than zero generation with more clovs, but still lighter compared with
control. It is possible that second generation has no difference with control or surpass control.
引文
1 Ploaie PG. Use of immune electron microscopy o detect viruses in plants regenerated from meristem tip cultures. Buletinul de Protectia Plantlor, 1984,
2 徐培文，孙慧生，孙瑞杰．脱毒大蒜速繁途径探讨和良繁体系的建立．中国蔬菜，1993，5：9～13
3 李昌华，李小川，赵美华．大蒜茎尖脱毒技术及组织培养研究．华北农学报，1995，10(3)：20～25
4 Choi SL, Paek KY, Kwun KC. Effect of explant source and shoot trimming on shoot multiplication and bulb formation of garlic in vitro. Journal of the Korean Society for Horticultural Science, 1985, 26(4):304～312
5 赵顺庆，李鹏飞，范怀忠．花叶病大蒜的茎端组织脱毒培养．华南农业大学学报，1987，8(4)：1～8
6 Hernandez PR, Prado O, Gil DV. Production of virus-free garlic (Allium sativum L.) by meristem culture. Centro Agricola. 1994, 21(1):76～84
7 Camara FAA, Pasqual M, Pinto JEBP. Obtaining Plants from garlic (Allium sativum L.)cultivars Chinese and Chonan using in vitro cultured meristems. Ciencia e Pratica, 1989, 13(1):97～102
8 陈世儒，黄菊辉．大蒜离体快繁及脱毒．园艺学报，1991，3(3)：133～137
9 陆关成，李方．余姚白蒜组织培养脱毒的研究．浙江农业，学报，1991，3(3)：133～137
10 栾雨时，栾非时，崔喜波．大蒜茎尖培养脱毒效果．北方园艺，1996(4)：37
11 张明厚，魏培文，朱俊华．大蒜茎尖组培苗的检测．东北农业大学学报，1999，30(2)：105～110
12 Verbeek M, van Dijk P. van Well PMA. Efficiency of eradication of four viruses from garlic (Allium sativum L.) by meristem tip culture. European Journal of Plant Pathology 1995, 101:3, 231～239
13 丁辛顺．脱毒大蒜苗的生产检测技术．上海农业学报，1998，4(1)：23～28
14 Walkey DGA, Webb MJW, Bolland CJ, Miller A. Production of virus-free garlic (Allium sativum L.) and shallot (A. ascalonicum L.) by meristem-tip culture. J Hortic Science, 1987, 62(2):211～220
15 Luciani GF, Cafrune EE, Curvetto NR. Obtaintion of virus-free plant of red garlic((Allium sativum L.). Fitopatologia. 1998, 33(3): 165～169
16 Conci VC, Nome SF. Virus free garlic (Allium sativum L.) plant obtained by thermotherapy and meristem tip culture. Joural of Phytopathology. 1991, 132(3): 186～192
17 Tsuneyoshi T, Sumi S. Differentiation among garlic viruses in mixed infections based on RT-PCR procedures and direct tissue blotting immunoassays. Phytopathology, 1996, 86(3):253～259
18 Vcman R, Zel J, Ravnikar M. Thermotherapy in virus elimination from garlic: influences on shoot multiplication from meristems and bulb formation in vitro. Scientia Horticulturae, 1998,
73(4) : 193-202
19 高山林,金雍安,蔡朝晖。大蒜分生组织培养脱病毒和快速繁殖技术。植物资源与环境学报,2000, 9(3) : 15-18
20 Conci VC, moriconi DN, Nome SF. In vitro plantlet regeneration from callus in garlic(Allium sativum L). Plant Physiology, 1987, 83:4
21 陈典。大蒜脱毒种苗培养技术研究。北方园艺. 1997(2) :29-30
22 Yun Jong Sun, Hwang SeGu, Song InGyu. Mass multiplication of shoot though shoot-tip culture of garlic.
23 Ma Y, Wang HL, Zhang CJ. High rate of virus-free plantlet regeneration via garlic scape-tip culture. Plant Cell Reports, 1994, 14(1) :65-68
24 ## 19(3) :145-148
25 Yang SG, Lee HS, Jeong WJ, Min SR. Production of virus-free microbulbs of garlic (Allium sativum L.)by in vitro culture of vegetative and floral buds in immature involucre. Journal of the Korean Society for Horticulture Science, 1993, 34(3) : 179-183
26 M Ebi, N Kasai, K Masuda. Small inflorescence bulbils are best for micropropagation and virus elimination in garlic. Hort. Science, 2000, 35(4) :735-737
27 Laplitan VPC, Patena LF. Bulblet formation in vitro, a new approach to garlic (Allium sativum L.) "basic seed" production. Philippine Journal of Crop Science, 1992, 17(2) :89-94
28 Kopnova SI, Kosova AI. Proliferation of axillary and adventious shoots of garlic in vitro. Izvestiya Akademij Nauk Mddavskoi SSR, Biologicheskikh i khimicheskikh Nauk, 1986(6) :29-33
29 Nagakubo T, Nagasawa A, Ohkawa H. Micropropagation of garlic through in vitro bulblet fonuation. Plant Cell Tissue and Organ Culture, 1993, 32(2) :175-183
30 Conci VC, Moriconi DN, Nome SF. Apical meristem culture of six garlic (Allium sativum L.) clones. Phyton, Argentina, 1986, 46(2) : 187-194
31 Bovo OA, Mroginski LA. Obtaining garlic (Allium sativum L.) plants by in vitro meristem culture. Phyton, Argentia, 1985, 45(2) 159-163
32 Masuda K, Hatakeyama E, Ito A. Micropropagation of garlic (Allium sativum L.). Bulletin of the Akita Prefectural College of Agriculture, 1994, No.20,43-48
33 Suh SK, Park HG. Plant regeneration from the culture of garlic root explants. Jourmal of the Korean
Society for Horticultural Science, 1995,36(1) :31-37
34 Mohamed Yasseen Y, Splittstoesser WE, Litz RE. In vitro shoot proliferation and production of sets from garlic and shallot. Plant Cell Tissue and Organ Culture, 1994,36(2) :243-247
35 Haque MS, Wada T, Hattori K. Novel method of rapid micropropagation using cyclic bulblet formation from root tip explants in garlic. Breeding Science, 1998,48(3) 293-299
36 Choi SY, Oh JY, Kim JS. Meristem culture of galic (Allium sativum L.) 1． Effects of growth regulators through in vitro multi-propagation of garlic (Allium sativum L.). Research Reports of the Rural Development Administration, Biotechnology, 1992,34(2) :24-29
37 Matsubara S, Chen D, Masuda M. The production and proliferation of bulbs from receptacles of virus-free garlic plants. Scientific Reports of the Faculty of Agriculture, 1990, No 75, 9-13
38 Havel L, Novak FJ. Morphogenic ability of in vitro cultured flower heads in Allium. Acta Universitatis Agriculturae Brno, 1985, 33(3) :615-617
39 Rauber M, Grunewaldt J. In vitro regeneration in Allium species. Plant Cell Reports, 1988, 7(6) :426-429
40 Barringer SA, Mohamed Yasseen Y; Schloupt RM. Regeneration of Allium Spp, in vitro by slicing the basal plate. Journal of Vegetable Crop Production, 1996,2(1) :27-33
41 Ayabe M, Sumi S. Establishment of a novel tissue culture method, stem-disc cuture, and its practical application to micropropagation of garlic (Allium sativum L.). Plant Cell Reports, 1998, 17(10) : 773-779
42 Kudou R, Fujime Y, Amimoto K. Effects of plant growth regulators and sampling positions on organ formation of garlic.Technical Bulletin of the Faculty of Agriculture, Kagawa University, 1995, 47(1) : 15-22
43 薛万新,陆帼一。大蒜蒜瓣离体繁殖研究。西北农业学报,1994,3(3) :62-66
44 Haque M S, Wada T, Hattorik. Garlic roots for micropropagation through in vitro bulblet formation. Acta Horticulturae, 2000, No.520,45-52
45 Lassocinski W, Gorecka K, Gorecki R. Preliminary report on onion (Allium cepa L.) and garlic (Allium sativum L.) propagation in vitro. Bulletin of the Polish Academy of Sciences, Biological Sciences, 1985,33(1/6) :23-26
46 Munozc, Escaff M. Multiplication of garlic (Allium sativum L.) from bulbils obtained in vitro.
Simiente, 1987, 57(3): 106
47 吕启愚，周维燕，董雅一．从大蒜嫩叶诱导愈伤组织及植株再生．园艺学报，1982，9(2)：67～69
48 薛万新，陆帼一．大蒜贮藏叶再生鳞茎．植物生理学通讯，1992，28(1)：52
49 Seabrook J E A. In vitro propagation and bulb formation of garlic. Canadian Journal of Plant Science, 1994, 74(1):155～158
50 Mohamed Yasseen Y, Barringer S A, Splittstoesser W E.In vitro bulb production from Allium ssp. In Vitro Celluar and Developmental Biology Plant, 1995, 31 (1):51～52
51 Bertaccini A, Marani F, Borgia M. Shoot-tip culture of different garlic lines for virus elimination,Rivista della Crtofrutticoltura Italiana, 1986, 70(2):96～105
52 薛万新，陆帼一，杨竹莹．大蒜蒜瓣离体繁殖过程中的影响因素．西北农业大学学报，1996，24(3)：114～117
53 Lee E M, Ra S W, Lee J Y. Min S R.The effect of growth regulators and the conditions of stock bulb storage on callus growth of garlic in vitro. Research Reports of the Rural Development Administration, Horticulture, 1988, 30(3):90～95
55 刘高琼，李式军，张学平．大蒜试管鳞茎微繁技术研究．南京农业大学学报，1996，19(3)：31～36
56 刘廷明，刘文英，曹鸣庆，周桂珍．脱毒仓山大蒜原种繁育体系的建立及配套栽培技术，山东农业科学，1991(6)：3～5
57 曾淑冰．大蒜组织培养的研究．中国蔬菜，1982(3)：26～27
58 栾非时，陈典，陈友．脱毒大蒜花原始体培养增殖技术的研究．中国蔬菜，1995(3)：4～6
59 栾非时，陈典，崔喜波．大蒜茎尖愈伤组织的诱导、植株分化及变异的研究．北方园艺，1993(4)：23～25
60 杨乃博．大蒜全展叶愈伤组织的诱导和植株再生．植物生理学通讯，1981(6)：47～48
61 Akitsu Nagasawa, John J Finer. Induction of morphogenic callus cultures from leaf tissue of garlic. Hort Science, 1988, 23(6): 1068～1070
62 郭洪云，樊治成，李红蓉，傅连海．山东农业大学学报，1999，30(2)：183～185
63 王洪隆，康玉庆，张存金．大蒜花梗组织培养再生植株．华北农学报，1992，7(3)：66～70
64 张忠新，李仁敬．大蒜组织培养中胚状体的发生．实验生物学报，1983，16(3)：242～243
65 Xu PW, Sun RJ, Yang YJ.Strategy for the use of virus-free seed garlic in field production. Acta
Hortic, 1994, 358:307～311
66 陈典，栾非时，陈友．大蒜脱毒苗扩繁途径的探讨．北方园艺，1994(5)：16～17
67 Koch M, Tanami Z, Salomon R. Improved regeneration of shoots from garlic callus. HortScience, 1995, 30(2):378
68 徐培文，孙慧生，孙瑞杰，张培歧．大蒜茎尖培养脱毒及增产效果的研究．山东农业科学，1991，6：6～10
69 李志勇，郭勇，罗焕亮．大蒜愈伤组织诱导条件初报．广西植物，1999，19(3)：251～254
70 郑海柔．γ射线对大蒜(Allium sativum L.)叶片愈伤组织生长和幼苗分化的影响．上海农业学报，1997，13(2)：23～26
71 Kudou R, Fujime Y, Komatsu Y, Fukada N. Effects of plant growth regulators and cold pre-storage of bulb on callus formation of garlic. Technical Bulletin of the Faculty of Agriculture, Kagawa University, 1995, 47(2):96～106
72 Suh SangKi, Park HyoGwen, Suh SK, Park HG. Effects of temperature pretreatment, growth regulators and antibiotic treatments on anther cultures of various cultivars of garlic (Allium sativum L.). Journal of the Korean Society for Horticultural Science, 1994, 35(4):337～344
73 王洪隆，康玉庆，张存金．大蒜发芽叶培养体系胞胚发生．华北农学报，1994，9(1)：92～94
74 Sushma Koul, Manju Sambyal, Grewal S, Koul S. Embryogenesis and plantlet formation in garlic Allium sativum L. Journal of Spices and Aromatic Crops, 1994, 3(1):43～47
75 Suh SK, Park HG. Rapid multiplication through immature bulbil culture of garlic. Journal of the Korean Society for Horticultural Science, 1993, 34(3): 173～178
76 Xue HM, Araki H, Shi L, Yakuwa T Somatic embryogenesis and plant regeneration in basbal plate and receptacle derived-callus cultures of garlic (Allium sativum L.) Journal of the Japanese Society for Horticutural Science, 1991, 60(3):627～634
77 Shuto H, Abe T, Sasahara T. In vitro propagation of plants from root apet-derived calli in Chinese Chive (Allium tuberosum Rottler) and garlic (Allium sativum L.). Japan. J Breed, 1993,43:349～354
78 Choi SY, Pack KY, Jo JT Plantlet production through callus culture in Allium sativum L. Journal of the Korean Society for Horticultural Science, 1993, 34(1): 16～28
79 Maggioni L, Cordi C, Fogher C. Callus induction, ploidy level and plant regeneration in in vitro garlic (Allium sativum L.) culcures. Journal of Genetics and Breeding, 1989, 43(4):251～254
80 Moriconi DN, Conci VC, Nome SF. In vitro plantlet regeneration from callus in garlic (Allium sativum L.). Phyton Buenos Aires, 1989, 49(1-2) :97-103
81 Nagasawa A, Finer IJ. Induction of morphogenic callus cultures from leaf tissue of garlic. HortScience, 1988, 23:6, I ,1068-10709
82 Suh SK, Park HG. Studies on the anther culture of garlic (Allium sativum L.) I Callus formation and plant regeneration. Journal of the Korean Society for Horticultural Science, 1986, 27(2) :89-95
83 Tapia MI. Meristematic root tumours: a new explant obtained by in vitro micropropagation of garlic (Allium sativum L.).Simiente, 1987, 57(3) : 104
84 Marchesi G, Corti C. An assessment of the effect of type of sugar on callus formation in garlic explains. Semeti Elette, 1986, 32(6) :40
85 Myers M, Simon P. Callus induction in garlic and its implications for transformation. Hort Science, 1993, 28:498
86 Marchesi G, Maggioni L. In vitro induction of callus from different explants of garlic bubils (Allium sativum L.). Sementi Elette. 1985, 31(5) :9-12
87 Tapia MI. Organogenesis and in vitro regeneration of garlic (Allium sativum L.). Plant , Simiente, 1987, 57(3) : 103-104
88 Koch M, Salomon R.Improment of garlic via somaclonal variation and virus elimination. Acta Hortic, 1994,358:211-214
89 Choi SY, Paek KL, Jo JT. Plantlet production through callus culture in (Allium sativum L.).
90 Tapia MI. Callus induction,organogensis and in vitro plant regeneration of garlic (Allium sativum L.). Agro Ciencia, 1996, 12(2) : 149-154
91 saker MM, Sawahel WA. Origin of garlic explant govern their competence for plant regeneration. Bulletin of the National Research Centre Cairo, 1998, 23(2) :203-211
92 曾淑冰。从蒜瓣再生完整植株。植物生理学通讯, 1981,4:45
93 郑海柔。大蒜试管苗中鳞茎的诱导。上海农业科技, 1991(2) :56
94 Dolezel J, Novak FJ. Cytogenetic effect of plant tissue culture medium with certain growth substances on Allium sativum L. Meristem root tip cells.
95 Dolezel J, Novak FJ. Karyological and cytophotometric study of callus induction in Allium sativum L. Journal of Plant Physiology, 1985, 118(5) :421-429
96 Resende FV, Souza RJ, Pasqual M. Field performation of garlic clones obtained by meristemtip culture. Horticultra Brasileira, 1995, 13(1) :44-46
97 Cihalikova J, Dolezel J. Application of the method of differential staining of sister chromatids in the cytogenetics of explant cultures. Sbornik UVTIZ, Genetika a Slechteni, 1986, 22:3,245
98 Dolezel J, Novak FJ. Sister chromatid exchanges in garlic(Allium sativum L.) callus cells. Plant Cell Reports, 1986, 5(4) :280-283
99 Dolezel J, Novak FJ. Karyological changes in the course of dedifferatiation of garlic(Allium sativum L.)cells. Kul tura kletok rastenii i biotekhnologiya, 1986,20-25
100 Novak FJ. Somaclinal variation in garlic tissue culture as a new breeding system, 3rd Eucarpia Allium Symposium, Wageningen, 4-6, September 1984, 39-43
101 Fogher C. Corti C, Savazzini F, Delledonne M. Obtaining virus-free material in garlic somaclones. Sementi Elette, 1993,39(1) : 13-16
102 Kwon YS, Lee HS, Yoon JI, Kim CB. Effects of planting density on reduction of secondary growth of virus-free garlic derived from apical meristem culture. J. Korea Soc Hort Sci, 1995, 36(4) :473-480
103 Park CH, Lee MS. Choi DC. Lim HC.Effect of naphthaleneacetic acid [NAA] and sucrose on bulblet formation in floral and vegetativebud culture of garlic(Allium sativum L.). J. Korea Soc Hort Sic, 1993, 34(4) :248-256
104 Lammerink. Quality Improvments in Printanor garlic. New Zealand Commercial Grower, 1989, 44(4) : 18-19
105 Walkey DGA, Antill DN. Agronomic evalution of virus-free and virus-infected garlic(Allium sativum L.). Journal of Horticutural Science, 1989, 64(1) :53-60
106 Takagi H, Qu Y, Kozai T. Zimmerman RH. Effect of light quality, photoperiod and cold treatment on in vitro bulbing of garlic shoot tip. Acta Horticulturae, 1995,No.393,181-188
107 Matsubara S, Chen D. In vitro production of garlic plants and field acclimatization. Hort Science, 1989,24(4) :677-679
108 Fogher C, Maggioni L, Corti C. Effects of inoculation with Glomus mosseae on rooting of plantlets of garlic(Allium sativum L.) regenerated from callus. Annali della Facoltadi Agraria, Universita Cattolica del Sacro Cuore, Molano,1986, 26(1) :9-14
109 Seabrovk JEA. In vitro propagation and bulb formation of garlic. Candian Journal of Plant Science, 1994, 74(1):155～158
110 徐培文，马伟青．大蒜组织培养材料染色体倍性变化研究．山东农业科学，1998，5：30～31
111 Maggioni L, Marchesi G. Plantlets of garlic(Allium sativum L.) obtained in vitro, Sementi Elette, 1984, 30(5):29～31
112 Pierik RLM. In vitro culture of higher plant. 1987. Martinus Nijhoff Publishers, Dordrecht.
113 曹静，周丽侬，邝哲师．影响香芋不定芽增殖因素探讨．广东农业科学，1998，2：16～17
114 刘选明，周朴华，屈妹妹．百合鳞片叶离体诱导形成不定芽和体细胞胚．园艺学报，1997，24(4)：353～358
115 师校欣，杜国强，高仪．苹果离体叶片高效再生不定芽技术研究．果树科学，1999，16(4)：255～258
116 孔令汶，谭智达，卞元生，郑淑湘．影响桑树叶片培养不定芽形态分化主要因素的研究．蚕业科学，1995，21(2)：67～71
117 周音，张智齐，钟维瑾．3种生长调节物质对生菜不定芽直接再生的影响．吉林农业大学学报，1998，20(3)：31～34
118 张初贤，余建明，蔡小宁，朱卫民．2，4-D对小白菜和大白菜子叶诱导不定芽的影响．江苏农业研究，1999，20(4)：75～76
119 于小林，李乃坚，黄自然，李颖．辣椒子叶离体培养和植株再生体系的建立．园艺学报，2000，27(1)：42～46
120 赵泓，刘凡，姚磊．简单快捷建立高频黄瓜子叶离体再生体系．生物技术，2000，10(2)：9～11
121 张智齐，周音，张玉华，钟维瑾．小白菜子叶诱导不定芽再生植株．上海农业学报，1998，14(2)：25～28
122 石淑稳，周光明，王新发．影响油菜子叶外植体不定芽高频率再生的因素．西北植物学报，1998，18(4)：477～482
123 张一凡．香蕉组织培养获取试管苗的简易方法及影响不定芽发生因素的研究．云南教育学院学报，1996，12(2)：70～73
124 石淑稳，周永明．甘蓝型油菜下胚轴培养和高频率芽再生技术的研究．中国油料作物学报，1998，20(2)：1～6
125 Radke SE, Turner JC. Transformation and regeneration of Brassica rapa using Agrobacterium
tumefaciens. Plant Cell Reports, 1992, 11: 499-505
126 Evans D A, Sharp W R. Single gene mutation in tomato plants regenerated from tissue culture. Science, 1983,221,947-951
127 Islam R, Hossein M, Reza MA, Mamun ANK, Joarder 01． Adventitious shoot regeneration from root tips of intact seedings of Aegle marmelos. Journal of Horticultural Science, 1996 71(6) :995-1000
128 Lee EM, Kim HS, Moon CS. Microprogation of high quality garlic (Allium sativum L.) through involucre culture. Abst, Symp, X XIV th Intern. Hort. Conf., 176
2 徐培文，孙慧生，孙瑞杰．脱毒大蒜速繁途径探讨和良繁体系的建立．中国蔬菜，1993，5：9～13
3 李昌华，李小川，赵美华．大蒜茎尖脱毒技术及组织培养研究．华北农学报，1995，10(3)：20～25
4 Choi SL, Paek KY, Kwun KC. Effect of explant source and shoot trimming on shoot multiplication and bulb formation of garlic in vitro. Journal of the Korean Society for Horticultural Science, 1985, 26(4):304～312
5 赵顺庆，李鹏飞，范怀忠．花叶病大蒜的茎端组织脱毒培养．华南农业大学学报，1987，8(4)：1～8
6 Hernandez PR, Prado O, Gil DV. Production of virus-free garlic (Allium sativum L.) by meristem culture. Centro Agricola. 1994, 21(1):76～84
7 Camara FAA, Pasqual M, Pinto JEBP. Obtaining Plants from garlic (Allium sativum L.)cultivars Chinese and Chonan using in vitro cultured meristems. Ciencia e Pratica, 1989, 13(1):97～102
8 陈世儒，黄菊辉．大蒜离体快繁及脱毒．园艺学报，1991，3(3)：133～137
9 陆关成，李方．余姚白蒜组织培养脱毒的研究．浙江农业，学报，1991，3(3)：133～137
10 栾雨时，栾非时，崔喜波．大蒜茎尖培养脱毒效果．北方园艺，1996(4)：37
11 张明厚，魏培文，朱俊华．大蒜茎尖组培苗的检测．东北农业大学学报，1999，30(2)：105～110
12 Verbeek M, van Dijk P. van Well PMA. Efficiency of eradication of four viruses from garlic (Allium sativum L.) by meristem tip culture. European Journal of Plant Pathology 1995, 101:3, 231～239
13 丁辛顺．脱毒大蒜苗的生产检测技术．上海农业学报，1998，4(1)：23～28
14 Walkey DGA, Webb MJW, Bolland CJ, Miller A. Production of virus-free garlic (Allium sativum L.) and shallot (A. ascalonicum L.) by meristem-tip culture. J Hortic Science, 1987, 62(2):211～220
15 Luciani GF, Cafrune EE, Curvetto NR. Obtaintion of virus-free plant of red garlic((Allium sativum L.). Fitopatologia. 1998, 33(3): 165～169
16 Conci VC, Nome SF. Virus free garlic (Allium sativum L.) plant obtained by thermotherapy and meristem tip culture. Joural of Phytopathology. 1991, 132(3): 186～192
17 Tsuneyoshi T, Sumi S. Differentiation among garlic viruses in mixed infections based on RT-PCR procedures and direct tissue blotting immunoassays. Phytopathology, 1996, 86(3):253～259
18 Vcman R, Zel J, Ravnikar M. Thermotherapy in virus elimination from garlic: influences on shoot multiplication from meristems and bulb formation in vitro. Scientia Horticulturae, 1998,
73(4) : 193-202
19 高山林,金雍安,蔡朝晖。大蒜分生组织培养脱病毒和快速繁殖技术。植物资源与环境学报,2000, 9(3) : 15-18
20 Conci VC, moriconi DN, Nome SF. In vitro plantlet regeneration from callus in garlic(Allium sativum L). Plant Physiology, 1987, 83:4
21 陈典。大蒜脱毒种苗培养技术研究。北方园艺. 1997(2) :29-30
22 Yun Jong Sun, Hwang SeGu, Song InGyu. Mass multiplication of shoot though shoot-tip culture of garlic.
23 Ma Y, Wang HL, Zhang CJ. High rate of virus-free plantlet regeneration via garlic scape-tip culture. Plant Cell Reports, 1994, 14(1) :65-68
24 ## 19(3) :145-148
25 Yang SG, Lee HS, Jeong WJ, Min SR. Production of virus-free microbulbs of garlic (Allium sativum L.)by in vitro culture of vegetative and floral buds in immature involucre. Journal of the Korean Society for Horticulture Science, 1993, 34(3) : 179-183
26 M Ebi, N Kasai, K Masuda. Small inflorescence bulbils are best for micropropagation and virus elimination in garlic. Hort. Science, 2000, 35(4) :735-737
27 Laplitan VPC, Patena LF. Bulblet formation in vitro, a new approach to garlic (Allium sativum L.) "basic seed" production. Philippine Journal of Crop Science, 1992, 17(2) :89-94
28 Kopnova SI, Kosova AI. Proliferation of axillary and adventious shoots of garlic in vitro. Izvestiya Akademij Nauk Mddavskoi SSR, Biologicheskikh i khimicheskikh Nauk, 1986(6) :29-33
29 Nagakubo T, Nagasawa A, Ohkawa H. Micropropagation of garlic through in vitro bulblet fonuation. Plant Cell Tissue and Organ Culture, 1993, 32(2) :175-183
30 Conci VC, Moriconi DN, Nome SF. Apical meristem culture of six garlic (Allium sativum L.) clones. Phyton, Argentina, 1986, 46(2) : 187-194
31 Bovo OA, Mroginski LA. Obtaining garlic (Allium sativum L.) plants by in vitro meristem culture. Phyton, Argentia, 1985, 45(2) 159-163
32 Masuda K, Hatakeyama E, Ito A. Micropropagation of garlic (Allium sativum L.). Bulletin of the Akita Prefectural College of Agriculture, 1994, No.20,43-48
33 Suh SK, Park HG. Plant regeneration from the culture of garlic root explants. Jourmal of the Korean
Society for Horticultural Science, 1995,36(1) :31-37
34 Mohamed Yasseen Y, Splittstoesser WE, Litz RE. In vitro shoot proliferation and production of sets from garlic and shallot. Plant Cell Tissue and Organ Culture, 1994,36(2) :243-247
35 Haque MS, Wada T, Hattori K. Novel method of rapid micropropagation using cyclic bulblet formation from root tip explants in garlic. Breeding Science, 1998,48(3) 293-299
36 Choi SY, Oh JY, Kim JS. Meristem culture of galic (Allium sativum L.) 1． Effects of growth regulators through in vitro multi-propagation of garlic (Allium sativum L.). Research Reports of the Rural Development Administration, Biotechnology, 1992,34(2) :24-29
37 Matsubara S, Chen D, Masuda M. The production and proliferation of bulbs from receptacles of virus-free garlic plants. Scientific Reports of the Faculty of Agriculture, 1990, No 75, 9-13
38 Havel L, Novak FJ. Morphogenic ability of in vitro cultured flower heads in Allium. Acta Universitatis Agriculturae Brno, 1985, 33(3) :615-617
39 Rauber M, Grunewaldt J. In vitro regeneration in Allium species. Plant Cell Reports, 1988, 7(6) :426-429
40 Barringer SA, Mohamed Yasseen Y; Schloupt RM. Regeneration of Allium Spp, in vitro by slicing the basal plate. Journal of Vegetable Crop Production, 1996,2(1) :27-33
41 Ayabe M, Sumi S. Establishment of a novel tissue culture method, stem-disc cuture, and its practical application to micropropagation of garlic (Allium sativum L.). Plant Cell Reports, 1998, 17(10) : 773-779
42 Kudou R, Fujime Y, Amimoto K. Effects of plant growth regulators and sampling positions on organ formation of garlic.Technical Bulletin of the Faculty of Agriculture, Kagawa University, 1995, 47(1) : 15-22
43 薛万新,陆帼一。大蒜蒜瓣离体繁殖研究。西北农业学报,1994,3(3) :62-66
44 Haque M S, Wada T, Hattorik. Garlic roots for micropropagation through in vitro bulblet formation. Acta Horticulturae, 2000, No.520,45-52
45 Lassocinski W, Gorecka K, Gorecki R. Preliminary report on onion (Allium cepa L.) and garlic (Allium sativum L.) propagation in vitro. Bulletin of the Polish Academy of Sciences, Biological Sciences, 1985,33(1/6) :23-26
46 Munozc, Escaff M. Multiplication of garlic (Allium sativum L.) from bulbils obtained in vitro.
Simiente, 1987, 57(3): 106
47 吕启愚，周维燕，董雅一．从大蒜嫩叶诱导愈伤组织及植株再生．园艺学报，1982，9(2)：67～69
48 薛万新，陆帼一．大蒜贮藏叶再生鳞茎．植物生理学通讯，1992，28(1)：52
49 Seabrook J E A. In vitro propagation and bulb formation of garlic. Canadian Journal of Plant Science, 1994, 74(1):155～158
50 Mohamed Yasseen Y, Barringer S A, Splittstoesser W E.In vitro bulb production from Allium ssp. In Vitro Celluar and Developmental Biology Plant, 1995, 31 (1):51～52
51 Bertaccini A, Marani F, Borgia M. Shoot-tip culture of different garlic lines for virus elimination,Rivista della Crtofrutticoltura Italiana, 1986, 70(2):96～105
52 薛万新，陆帼一，杨竹莹．大蒜蒜瓣离体繁殖过程中的影响因素．西北农业大学学报，1996，24(3)：114～117
53 Lee E M, Ra S W, Lee J Y. Min S R.The effect of growth regulators and the conditions of stock bulb storage on callus growth of garlic in vitro. Research Reports of the Rural Development Administration, Horticulture, 1988, 30(3):90～95
55 刘高琼，李式军，张学平．大蒜试管鳞茎微繁技术研究．南京农业大学学报，1996，19(3)：31～36
56 刘廷明，刘文英，曹鸣庆，周桂珍．脱毒仓山大蒜原种繁育体系的建立及配套栽培技术，山东农业科学，1991(6)：3～5
57 曾淑冰．大蒜组织培养的研究．中国蔬菜，1982(3)：26～27
58 栾非时，陈典，陈友．脱毒大蒜花原始体培养增殖技术的研究．中国蔬菜，1995(3)：4～6
59 栾非时，陈典，崔喜波．大蒜茎尖愈伤组织的诱导、植株分化及变异的研究．北方园艺，1993(4)：23～25
60 杨乃博．大蒜全展叶愈伤组织的诱导和植株再生．植物生理学通讯，1981(6)：47～48
61 Akitsu Nagasawa, John J Finer. Induction of morphogenic callus cultures from leaf tissue of garlic. Hort Science, 1988, 23(6): 1068～1070
62 郭洪云，樊治成，李红蓉，傅连海．山东农业大学学报，1999，30(2)：183～185
63 王洪隆，康玉庆，张存金．大蒜花梗组织培养再生植株．华北农学报，1992，7(3)：66～70
64 张忠新，李仁敬．大蒜组织培养中胚状体的发生．实验生物学报，1983，16(3)：242～243
65 Xu PW, Sun RJ, Yang YJ.Strategy for the use of virus-free seed garlic in field production. Acta
Hortic, 1994, 358:307～311
66 陈典，栾非时，陈友．大蒜脱毒苗扩繁途径的探讨．北方园艺，1994(5)：16～17
67 Koch M, Tanami Z, Salomon R. Improved regeneration of shoots from garlic callus. HortScience, 1995, 30(2):378
68 徐培文，孙慧生，孙瑞杰，张培歧．大蒜茎尖培养脱毒及增产效果的研究．山东农业科学，1991，6：6～10
69 李志勇，郭勇，罗焕亮．大蒜愈伤组织诱导条件初报．广西植物，1999，19(3)：251～254
70 郑海柔．γ射线对大蒜(Allium sativum L.)叶片愈伤组织生长和幼苗分化的影响．上海农业学报，1997，13(2)：23～26
71 Kudou R, Fujime Y, Komatsu Y, Fukada N. Effects of plant growth regulators and cold pre-storage of bulb on callus formation of garlic. Technical Bulletin of the Faculty of Agriculture, Kagawa University, 1995, 47(2):96～106
72 Suh SangKi, Park HyoGwen, Suh SK, Park HG. Effects of temperature pretreatment, growth regulators and antibiotic treatments on anther cultures of various cultivars of garlic (Allium sativum L.). Journal of the Korean Society for Horticultural Science, 1994, 35(4):337～344
73 王洪隆，康玉庆，张存金．大蒜发芽叶培养体系胞胚发生．华北农学报，1994，9(1)：92～94
74 Sushma Koul, Manju Sambyal, Grewal S, Koul S. Embryogenesis and plantlet formation in garlic Allium sativum L. Journal of Spices and Aromatic Crops, 1994, 3(1):43～47
75 Suh SK, Park HG. Rapid multiplication through immature bulbil culture of garlic. Journal of the Korean Society for Horticultural Science, 1993, 34(3): 173～178
76 Xue HM, Araki H, Shi L, Yakuwa T Somatic embryogenesis and plant regeneration in basbal plate and receptacle derived-callus cultures of garlic (Allium sativum L.) Journal of the Japanese Society for Horticutural Science, 1991, 60(3):627～634
77 Shuto H, Abe T, Sasahara T. In vitro propagation of plants from root apet-derived calli in Chinese Chive (Allium tuberosum Rottler) and garlic (Allium sativum L.). Japan. J Breed, 1993,43:349～354
78 Choi SY, Pack KY, Jo JT Plantlet production through callus culture in Allium sativum L. Journal of the Korean Society for Horticultural Science, 1993, 34(1): 16～28
79 Maggioni L, Cordi C, Fogher C. Callus induction, ploidy level and plant regeneration in in vitro garlic (Allium sativum L.) culcures. Journal of Genetics and Breeding, 1989, 43(4):251～254
80 Moriconi DN, Conci VC, Nome SF. In vitro plantlet regeneration from callus in garlic (Allium sativum L.). Phyton Buenos Aires, 1989, 49(1-2) :97-103
81 Nagasawa A, Finer IJ. Induction of morphogenic callus cultures from leaf tissue of garlic. HortScience, 1988, 23:6, I ,1068-10709
82 Suh SK, Park HG. Studies on the anther culture of garlic (Allium sativum L.) I Callus formation and plant regeneration. Journal of the Korean Society for Horticultural Science, 1986, 27(2) :89-95
83 Tapia MI. Meristematic root tumours: a new explant obtained by in vitro micropropagation of garlic (Allium sativum L.).Simiente, 1987, 57(3) : 104
84 Marchesi G, Corti C. An assessment of the effect of type of sugar on callus formation in garlic explains. Semeti Elette, 1986, 32(6) :40
85 Myers M, Simon P. Callus induction in garlic and its implications for transformation. Hort Science, 1993, 28:498
86 Marchesi G, Maggioni L. In vitro induction of callus from different explants of garlic bubils (Allium sativum L.). Sementi Elette. 1985, 31(5) :9-12
87 Tapia MI. Organogenesis and in vitro regeneration of garlic (Allium sativum L.). Plant , Simiente, 1987, 57(3) : 103-104
88 Koch M, Salomon R.Improment of garlic via somaclonal variation and virus elimination. Acta Hortic, 1994,358:211-214
89 Choi SY, Paek KL, Jo JT. Plantlet production through callus culture in (Allium sativum L.).
90 Tapia MI. Callus induction,organogensis and in vitro plant regeneration of garlic (Allium sativum L.). Agro Ciencia, 1996, 12(2) : 149-154
91 saker MM, Sawahel WA. Origin of garlic explant govern their competence for plant regeneration. Bulletin of the National Research Centre Cairo, 1998, 23(2) :203-211
92 曾淑冰。从蒜瓣再生完整植株。植物生理学通讯, 1981,4:45
93 郑海柔。大蒜试管苗中鳞茎的诱导。上海农业科技, 1991(2) :56
94 Dolezel J, Novak FJ. Cytogenetic effect of plant tissue culture medium with certain growth substances on Allium sativum L. Meristem root tip cells.
95 Dolezel J, Novak FJ. Karyological and cytophotometric study of callus induction in Allium sativum L. Journal of Plant Physiology, 1985, 118(5) :421-429
96 Resende FV, Souza RJ, Pasqual M. Field performation of garlic clones obtained by meristemtip culture. Horticultra Brasileira, 1995, 13(1) :44-46
97 Cihalikova J, Dolezel J. Application of the method of differential staining of sister chromatids in the cytogenetics of explant cultures. Sbornik UVTIZ, Genetika a Slechteni, 1986, 22:3,245
98 Dolezel J, Novak FJ. Sister chromatid exchanges in garlic(Allium sativum L.) callus cells. Plant Cell Reports, 1986, 5(4) :280-283
99 Dolezel J, Novak FJ. Karyological changes in the course of dedifferatiation of garlic(Allium sativum L.)cells. Kul tura kletok rastenii i biotekhnologiya, 1986,20-25
100 Novak FJ. Somaclinal variation in garlic tissue culture as a new breeding system, 3rd Eucarpia Allium Symposium, Wageningen, 4-6, September 1984, 39-43
101 Fogher C. Corti C, Savazzini F, Delledonne M. Obtaining virus-free material in garlic somaclones. Sementi Elette, 1993,39(1) : 13-16
102 Kwon YS, Lee HS, Yoon JI, Kim CB. Effects of planting density on reduction of secondary growth of virus-free garlic derived from apical meristem culture. J. Korea Soc Hort Sci, 1995, 36(4) :473-480
103 Park CH, Lee MS. Choi DC. Lim HC.Effect of naphthaleneacetic acid [NAA] and sucrose on bulblet formation in floral and vegetativebud culture of garlic(Allium sativum L.). J. Korea Soc Hort Sic, 1993, 34(4) :248-256
104 Lammerink. Quality Improvments in Printanor garlic. New Zealand Commercial Grower, 1989, 44(4) : 18-19
105 Walkey DGA, Antill DN. Agronomic evalution of virus-free and virus-infected garlic(Allium sativum L.). Journal of Horticutural Science, 1989, 64(1) :53-60
106 Takagi H, Qu Y, Kozai T. Zimmerman RH. Effect of light quality, photoperiod and cold treatment on in vitro bulbing of garlic shoot tip. Acta Horticulturae, 1995,No.393,181-188
107 Matsubara S, Chen D. In vitro production of garlic plants and field acclimatization. Hort Science, 1989,24(4) :677-679
108 Fogher C, Maggioni L, Corti C. Effects of inoculation with Glomus mosseae on rooting of plantlets of garlic(Allium sativum L.) regenerated from callus. Annali della Facoltadi Agraria, Universita Cattolica del Sacro Cuore, Molano,1986, 26(1) :9-14
109 Seabrovk JEA. In vitro propagation and bulb formation of garlic. Candian Journal of Plant Science, 1994, 74(1):155～158
110 徐培文，马伟青．大蒜组织培养材料染色体倍性变化研究．山东农业科学，1998，5：30～31
111 Maggioni L, Marchesi G. Plantlets of garlic(Allium sativum L.) obtained in vitro, Sementi Elette, 1984, 30(5):29～31
112 Pierik RLM. In vitro culture of higher plant. 1987. Martinus Nijhoff Publishers, Dordrecht.
113 曹静，周丽侬，邝哲师．影响香芋不定芽增殖因素探讨．广东农业科学，1998，2：16～17
114 刘选明，周朴华，屈妹妹．百合鳞片叶离体诱导形成不定芽和体细胞胚．园艺学报，1997，24(4)：353～358
115 师校欣，杜国强，高仪．苹果离体叶片高效再生不定芽技术研究．果树科学，1999，16(4)：255～258
116 孔令汶，谭智达，卞元生，郑淑湘．影响桑树叶片培养不定芽形态分化主要因素的研究．蚕业科学，1995，21(2)：67～71
117 周音，张智齐，钟维瑾．3种生长调节物质对生菜不定芽直接再生的影响．吉林农业大学学报，1998，20(3)：31～34
118 张初贤，余建明，蔡小宁，朱卫民．2，4-D对小白菜和大白菜子叶诱导不定芽的影响．江苏农业研究，1999，20(4)：75～76
119 于小林，李乃坚，黄自然，李颖．辣椒子叶离体培养和植株再生体系的建立．园艺学报，2000，27(1)：42～46
120 赵泓，刘凡，姚磊．简单快捷建立高频黄瓜子叶离体再生体系．生物技术，2000，10(2)：9～11
121 张智齐，周音，张玉华，钟维瑾．小白菜子叶诱导不定芽再生植株．上海农业学报，1998，14(2)：25～28
122 石淑稳，周光明，王新发．影响油菜子叶外植体不定芽高频率再生的因素．西北植物学报，1998，18(4)：477～482
123 张一凡．香蕉组织培养获取试管苗的简易方法及影响不定芽发生因素的研究．云南教育学院学报，1996，12(2)：70～73
124 石淑稳，周永明．甘蓝型油菜下胚轴培养和高频率芽再生技术的研究．中国油料作物学报，1998，20(2)：1～6
125 Radke SE, Turner JC. Transformation and regeneration of Brassica rapa using Agrobacterium
tumefaciens. Plant Cell Reports, 1992, 11: 499-505
126 Evans D A, Sharp W R. Single gene mutation in tomato plants regenerated from tissue culture. Science, 1983,221,947-951
127 Islam R, Hossein M, Reza MA, Mamun ANK, Joarder 01． Adventitious shoot regeneration from root tips of intact seedings of Aegle marmelos. Journal of Horticultural Science, 1996 71(6) :995-1000
128 Lee EM, Kim HS, Moon CS. Microprogation of high quality garlic (Allium sativum L.) through involucre culture. Abst, Symp, X XIV th Intern. Hort. Conf., 176