粉葛（Pueraria thomsonii Benth.）的组织培养及辐照对粉葛试管苗的作用的研究
|
摘要
粉葛是甘葛藤(Pueraria thomsonii Benth.)的干燥根,是一味常用中药。近年来,粉葛综合利用价值日益提高,已成为重要的药用植物和工业生产原料。本实验对粉葛快速繁殖体系及试管块根的诱导条件进行了研究,并探索了~(60)Coγ射线辐照对粉葛组培苗生长发育的影响。取得了以下结果:
1、粉葛试管快繁技术的研究
粉葛种子的休眠主要是种皮的不透性引起的,通过机械破坏种皮的方式能使种子的萌发率达到80%。采用75%酒精和0.1%升汞作为种子表面灭菌的灭菌剂,采用两次消毒的方法可将污染率降低到15%。试管苗的增殖培养基为:MS+6-BA2.0mg/L+NAA0.1mg/L ,增殖系数为18% ;试管苗壮苗培养基为:MS+6-BA0.1mg/L+NAA0.1mg/L;试管苗的生根培养基为:1/2MS+IBA1.0mg/L,生根率为90%。
2、粉葛试管块根诱导技术的研究
明确了粉葛试管块根诱导的最适培养基是:MS+6-BA1.5mg/L+NAA0.1mg/L。本研究发现较高的蔗糖含量和较高的温度能促进粉葛试管块根的增粗,最适宜的含糖量和温度分别为5%和25℃,试管块根直径可达0.6cm;添加外源GA3不利于试管块根形成。
3、~(60)Coγ射线辐照对粉葛试管苗生长的影响
随着辐照剂量的增大试管苗的生根受到抑制,在剂量率为1Gy/min的处理中,当剂量达到40Gy以上时,试管苗不再具备生根能力;当辐照剂量不超过10Gy时γ射线对试管苗的增高和增殖均有一定的促进作用,当剂量超过10Gy时,随着剂量的增大,试管苗的生长受到抑制,当剂量达到60Gy时,试管苗呈生长停滞状态;粉葛试管苗最适剂量是剂量率为1Gy/min条件下辐照计量为20Gy,致死剂量是剂量率为1Gy/min条件下辐照剂量为100Gy。
Radix Puerariae, the dry root of Pueraria thomsonii Benth., is one of traditional Chinese medicines. In recent years, Radix Puerariae powder, whose comprehensive value in use has been improved gradually became significant officinal and raw material for industry production. In this study, we focused our attention on the establishment of rapid propagation system and the induction condition of the tube earthnut, and also the effect of 60Coγray irradiation on the growth of cultured seedlings of Radix Puerariae. The results are listed as follow:
1. Study on the establishment of rapid propagation system of Radix Puerariae.
The dormancy of Radix Puerariae.seeds is caused by the imperviousness of seed coat, which can be smashed by machinery break and reached 80% of the germination rate . Sterilized by 75% ethanol and 0.1% HgCl2 for 2 times can reduce the pollution rate to 15%. Propagation culture medium of tube seedling is MS+6-BA2.0mg/L+NAA0.1mg/L, by which the grpgation coefficient was 18%; sound seedling culture medium is MS+6-BA0.1mg/L+NAA0.1mg/L; root inducing culture medium is 1/2MS+IBA1.0mg/L, and the root gemination rate was 90%.
2. Study on induction condition of the tube earthnut of Radix Puerariae.
The optimum tube earthnut induction culture medium is MS+6-BA1.5mg/L+NAA0.1mg/L; high sucrose content and higher temperature were both beneficial for the increase of the tube earthnut diameter., and the most suitable sucrose content and the optimum temperature for the tube earthnut induction were 5% and 25℃respectively; exogenous GA3 went against the formation of tube earthnut.
3. The impact of 60Coγray irradiation on the growth of Radix Puerariae tube seedlings.
The rooting ability was inhibited as the dose ofγray increased, when the dose was up to 40Gy and with the treatment of 1Gy/min the rooting ability of tube seedlings lose; and the dose lower than 10Gyγray can increase the height and propagation of tube seedling;when the irradiation dose surpass 10Gy the growth can be inhibited, and the dose upper to 60Gy can absolutely stop the growth of tube seedling; the optimum dose for the Radix Puerariae tube seedlings is 20Gy with the treatment of 1Gy/min, the fatal dose was 100Gy with the treatment of 1Gy/min.
1、粉葛试管快繁技术的研究
粉葛种子的休眠主要是种皮的不透性引起的,通过机械破坏种皮的方式能使种子的萌发率达到80%。采用75%酒精和0.1%升汞作为种子表面灭菌的灭菌剂,采用两次消毒的方法可将污染率降低到15%。试管苗的增殖培养基为:MS+6-BA2.0mg/L+NAA0.1mg/L ,增殖系数为18% ;试管苗壮苗培养基为:MS+6-BA0.1mg/L+NAA0.1mg/L;试管苗的生根培养基为:1/2MS+IBA1.0mg/L,生根率为90%。
2、粉葛试管块根诱导技术的研究
明确了粉葛试管块根诱导的最适培养基是:MS+6-BA1.5mg/L+NAA0.1mg/L。本研究发现较高的蔗糖含量和较高的温度能促进粉葛试管块根的增粗,最适宜的含糖量和温度分别为5%和25℃,试管块根直径可达0.6cm;添加外源GA3不利于试管块根形成。
3、~(60)Coγ射线辐照对粉葛试管苗生长的影响
随着辐照剂量的增大试管苗的生根受到抑制,在剂量率为1Gy/min的处理中,当剂量达到40Gy以上时,试管苗不再具备生根能力;当辐照剂量不超过10Gy时γ射线对试管苗的增高和增殖均有一定的促进作用,当剂量超过10Gy时,随着剂量的增大,试管苗的生长受到抑制,当剂量达到60Gy时,试管苗呈生长停滞状态;粉葛试管苗最适剂量是剂量率为1Gy/min条件下辐照计量为20Gy,致死剂量是剂量率为1Gy/min条件下辐照剂量为100Gy。
Radix Puerariae, the dry root of Pueraria thomsonii Benth., is one of traditional Chinese medicines. In recent years, Radix Puerariae powder, whose comprehensive value in use has been improved gradually became significant officinal and raw material for industry production. In this study, we focused our attention on the establishment of rapid propagation system and the induction condition of the tube earthnut, and also the effect of 60Coγray irradiation on the growth of cultured seedlings of Radix Puerariae. The results are listed as follow:
1. Study on the establishment of rapid propagation system of Radix Puerariae.
The dormancy of Radix Puerariae.seeds is caused by the imperviousness of seed coat, which can be smashed by machinery break and reached 80% of the germination rate . Sterilized by 75% ethanol and 0.1% HgCl2 for 2 times can reduce the pollution rate to 15%. Propagation culture medium of tube seedling is MS+6-BA2.0mg/L+NAA0.1mg/L, by which the grpgation coefficient was 18%; sound seedling culture medium is MS+6-BA0.1mg/L+NAA0.1mg/L; root inducing culture medium is 1/2MS+IBA1.0mg/L, and the root gemination rate was 90%.
2. Study on induction condition of the tube earthnut of Radix Puerariae.
The optimum tube earthnut induction culture medium is MS+6-BA1.5mg/L+NAA0.1mg/L; high sucrose content and higher temperature were both beneficial for the increase of the tube earthnut diameter., and the most suitable sucrose content and the optimum temperature for the tube earthnut induction were 5% and 25℃respectively; exogenous GA3 went against the formation of tube earthnut.
3. The impact of 60Coγray irradiation on the growth of Radix Puerariae tube seedlings.
The rooting ability was inhibited as the dose ofγray increased, when the dose was up to 40Gy and with the treatment of 1Gy/min the rooting ability of tube seedlings lose; and the dose lower than 10Gyγray can increase the height and propagation of tube seedling;when the irradiation dose surpass 10Gy the growth can be inhibited, and the dose upper to 60Gy can absolutely stop the growth of tube seedling; the optimum dose for the Radix Puerariae tube seedlings is 20Gy with the treatment of 1Gy/min, the fatal dose was 100Gy with the treatment of 1Gy/min.
引文
[1] 郑筱萸,蒋作君,邵明立,等.中华人民共和国药典.北京中科印刷有限公司,2005.203
[2] 李时珍.本草纲目.北京:人民卫生出版社(王育杰整理),1999,1151~1154
[3] 肖培根,李大鹏,黄世林,等.新编中药志,第一卷[M].北京:中国医药科技出版杜,2002:962~975
[4] 吴德邻,陈忠毅,黄向旭,等.中国葛属(Pueraria DC)的研究.热带亚热带植物学报,1994,2(3):12~21
[5] 张仁岗,刘福明.粉葛的心血管药理作用研究进展[J].中药材,2001,24(7):535~537
[6] 陶娟,许慕农,路秋生,等.中国葛属植物资源和利用情况.中国野生植物资源,2007,26(3):38~40
[7] 薛建平,柳俊,蒋细旺,等.药用植物生物技术.合肥:中国科学技术大学出版社,2005.1~3
[8] 李文安,余叔文.植物生理与分子生物学.北京:科学技术出版社,1992,37
[9] 翟中和,王喜忠,丁明孝,等.细胞生物学.北京:高等教育出版社,2000
[10] 谷瑞升,蒋湘宁,郭仲琛,等.植物离体培养中器官发生调控机制研究进展.植物学通报.1999,16(3):238~244
[11] 李明军,薛建平,陈明霞,等.不同因子对山药愈伤组织诱导的影响.广西植物,2000,20(2):156~160
[12] 黄潞,卫志明.不同基因型玉米的再生能力和胚性与非胚性愈伤组织 DNA 的差异.植物生理学报.1999,25(4):332~338
[13] Piagnani C,Bassi D.In vivo and in vitro propagagation of Sorbus spp.from juvenile material.Italus Hortus.2000,7(5):3~7
[14] 黄学森,焦定量,那丽,等.西瓜子叶离体培养获得再生植株.中国西瓜甜瓜.1994,(3):15~16
[15] Preece JE . A century of progress with vegetative plant propagation.HortScience.2003,38:1015~1025
[16] 罗凤霞,徐桂华,金丽丽,等.新铁炮百合微繁的研究.沈阳农业大学学报,2000,31(3):254~257
[17] 王刚,杜捷,李桂英,等.看周百合和野百合组织培养及快速繁殖研究.西北师范大学学报(自然科学版),2002,38(1):69~71
[18] 刘敏.花卉组织培养与工业化生产.北京:地质出版社.2002:44
[19] Wagner J,Kafka I.Effect of medium composition on in vitro germination of embryos of Fraxinus excelsior at different stages of development.Plant Physiology.1995,146:566~568
[20] 兰素缺,李光威,权书月,等.碳源组分及浓度对小麦花药培养和游离小孢子培养的影响.华北农学报.2002,17(增刊):92~97
[21] 丁世萍,严菊强,季道藩,等.糖类在植物组织培养中的效应.植物学报.1998,15(6):42~46
[22] 王爱民,刘文,傅中滇,等.不同碳源对红边朱蕉组培苗生长的影响.徐州示范大学学报.2003,21(3):76~78
[23] Hadrami EL,Dauac J.Effcets of growth regulators on polyamine content and peroxidase activity in Hebea brasilienses callus.Ann,Bot,1992,69:323~325
[24] 詹亚光,样传平.白桦愈伤组织的高效诱导和不定芽分化.植物生理学通讯.2002,38(2):117~114
[25] Martinelli L,Gianazza E.Biochemical changes during regeneration of sunflower (Helianthus annuus L.).Electrophoresis.1996,17(1):191~197
[26] Michalczuk L,Ribnicky DM,Cooke TJ,et al.Regulation of indole-3-aceticacid biosynthetic pathway in carrot cell cultures.Plant Physiol.1992,100:1346~1353
[27] Leshem B,Ronen R,Soudry E,et al.Cytokinin at a large range of concentration determines rates of polypeptide metabolism and regeneratiom in cultere melon cotyledons.Plant Physiol.1994,143:330~336
[28] Kozai T,Tanaka K,Jeong BR,et al.Effect of relative humidity in the culture vessel on the growth and shoot elongation of potato plantlets in vitro.Journal of the Japanese Society for Horticultural Science.1993,62(2):413~417
[29] Reed BM,Mentzer J,Tanpraser P,et al.Internal Bacterial contamination of micropropagated hazelnut:indentification and antibiotic treatment.Plant Cell.Tissue and organ Culture.1998,(52):53~55
[30] Kritzinger EM,Vanbvuuren RJ,Woodward B,et al.Elimination of external and internal contamination in rhizomes of Zantedeschia aethiopica with commercial fungicides and antibiotics.Plant Cell.Tissue and Organ Culture.1998,(52):61~65
[31] 高国训.植物组织培养中的褐变问题.植物生理学通讯.1999,35(6):501~506
[32] 刘兰芳."薄壳香"核桃组织培养中的褐化及防止措施研究.园艺学报.2002,29(2):171~172
[33] 姚洪军,罗晓芳.植物组织培养外植体河边的研究进展.北京林业大学学报.1999,21(3):78~84
[34] Dan H,Meredith CP.The influence of explant origin on tissue browning and shoot production in shoot tip cultures of grapevine.Journal of the American Society for Horicultural Science.1986,25:456~466
[35] 程家胜,史永中,张志云,等.苹果组织培养中的玻璃苗问题.植物生理学通讯.1991,(1):33~35
[36] 郝瑞庆,杨广东.大白菜试管苗玻璃化发生机制初探.中国农学通报.2002,18(3):45~47
[37] 丁运华,王鸿博.3 种应诉对梅菜试管苗玻璃化的影响.热带农业科学.2003,23(23):5~9
[38] 于树宏,李玲.野葛的组织培养和植株再生.植物资源与环境.1998,8(1):63~64
[39] 施和平.三裂叶野葛茎段和叶柄培养再生植株.中草药.2000,31(7):550~552
[40] 周堂英,李惠波,向素琼,等.粉葛组织培养及同源四倍体诱导.2005,36(8):1230~1233
[41] 吴丽芳,张素芳,蒋亚莲,等.粉葛的离体培养和无糖生根.2005,41(5):646~647
[42] 夏英武.作物诱变育种.北京:中国农业出版社.1997.84~91
[43] 刘军,核科学技术在农业领域的应用.核科学百年讲座.2001
[44] 徐冠仁.核农学导论.北京:原子能出版社.1995
[45] Charbaji TN.Effect of low doses of gamma irradiation on in vitro growth ofgrapevine.Plant Cell,Tissue and Organ Culture,1999,57:129~132
[46] 吴关庭等.小麦辐射育种研究进展.核农学通报.1993.14:189~191
[47] 韩月澎,陈秀兰,何震天,等.大麦辐射育种研究进展.大麦科学.2000.3:11~13
[48] 杨爱珠.玉米辐射诱变的研究和应用.核农学通报.1995,16:158~159
[49] 万贤国.辐射诱变在创造种质资源上的应用作物研究.1994.8:1~4
[50] Ahloowalia BS,Maluszynski M:Induced mutations-A new paradigm in plant breeding Euphytica,2001,118:167~173
[51] 毛炎麟.辐射的生物学效应.植物诱变育种学北京:中国农业出版社.1996.79~100
[52] 杨期和,尹小娟,叶万辉,等.硬实种子休眠的机制和解除方法.植物学通报.2006.23(1):108~118
[53] 孙仲序,王玉军,刘静,等.植物组织快繁滤纸纸桥生根新技术[J].山东农业大学学报(自然科学版),2002,33(3):257~263
[54] Edwin BH.Non-axenic plant tissue culture:possibilities and opportunities [J].Acta Horticulture,1990,280:1990
[55] Flowler MW,Wastson R.In proc.5th Ind.Cong [J].Plant Tissue and Cell Culture.1982,Tissue Culture pp 225
[56] Ewing EE.The role of hormones in potato(Solanum tuberosum L.)tuberization.In:Davies P J ed.Plant hormones physiology,biochemistry and molecular biology.(2nd ed).Dordrecht:Kluwer Academic Pub,1995,698~724
[57] Khuri S,Moorby J.Investigations into the role of sucrose in potato cv.Estima microtuber production in vitro.Ann of Bot,1995,75:295~303
[58] 连勇,邹颖,杨宏福,等.马铃薯试管薯发育机理研究-温度对试管薯形成的影响.马铃薯杂志,1996a,10:133~137
[59] 沈清景,叶贻勋,凌永胜,等.马铃薯试管薯诱导因素研究,福建农业学报,2001,16(1):54~56
[60] 柳俊,谢从华.马铃薯块茎发育机理及其基因表达.植物学通报,2001,18(5):531~539
[61] 蒙美莲,刘梦芸,门福义,等.赤霉素和脱落酸对马铃薯块茎形成的影响.马铃薯杂志,1994,8 (3):134~137
[62] Stefano P.Mutation induction and tissue culture in improving fruits.Plant Cell,Tissue and Organ Culture,2001,64:185~2101
[63] 徐华军,贺源辉,陈秀芳,等.60Coγ 射线对双低甘蓝型油菜的辐射效应.核农学报.1992.6:199~206
[64] 宋恒,王长泉,巩向忠,等.γ 射线辐照杜鹃试管苗诱发突变体的研究.核农学报.2003.17:347~349
[65] 李林光,孙清荣,贺桂香,等.苹果离体叶片 60Coγ 辐射对不定稍再生及芽生长的影响.山东农业科学.2004:36~38
[66] 翁伯琦,徐国忠,郑向丽,等.60Coγ 射线辐照圆叶决明子对其生物学特性的影响.核农学报.2004.15:197~200
[67] 宋淑敏,田玉杰,姬研茹,等. γ 射线辐射亚麻花药的研究初报.中国麻业.2004.14:162~176
[68] Rama N,Swamy UT,Praveen M.Induction of streptomycin-resistant plantlets insolanum surattense through in vitro mutagenesis.Plant cell,Tissue and Organ Culture,2005,80:201~207
[69] Amer IM , ElRassas HN . Utilization of gamma irradiation in barley breeding.Annals of Agricultural Science (Cairo),1994,39:257~268
[70] Datta SK.Chrysanthemum cultivars evolved by induced mutation at National Botanical Research Institute,Lucknow.The Chrysanthemum,44:72~75
[71] Forster BP,Pakniyat H,Macaulay M,et al.Variation in the leaf sodium content of Hordeum vulgare (barley) cultivar Maythorpe and its derived mutant cv . Golden Promise.Heredity,73:249~253
[72] Ahloowalia BS.In-vitro techniques and mutagenesis for the improvement of vegetatively propagated plant.In:Jain M S,Brar D S,Ahloowalia B S(Eds.).Somaclonal Variation and Induced Mutations in Crop Improvement.293~309.Kluwer Academic Publishers,Dordrecht,NL.1998
[73] 王文恩,张俊卫,包满珠,等.60Coγ 对野牛草干种子的刺激生长效应.核农学报.2005.19:191~194.
[74] 杨期和,尹小娟,叶万辉,等.硬实种子休眠的机制和解除方法.植物学通报.2006.23(1):108~118
[75] 孙仲序,王玉军,刘静,等.植物组织快繁滤纸纸桥生根新技术[J].山东农业大学学报(自然科学版),2002,33(3):257~263
[2] 李时珍.本草纲目.北京:人民卫生出版社(王育杰整理),1999,1151~1154
[3] 肖培根,李大鹏,黄世林,等.新编中药志,第一卷[M].北京:中国医药科技出版杜,2002:962~975
[4] 吴德邻,陈忠毅,黄向旭,等.中国葛属(Pueraria DC)的研究.热带亚热带植物学报,1994,2(3):12~21
[5] 张仁岗,刘福明.粉葛的心血管药理作用研究进展[J].中药材,2001,24(7):535~537
[6] 陶娟,许慕农,路秋生,等.中国葛属植物资源和利用情况.中国野生植物资源,2007,26(3):38~40
[7] 薛建平,柳俊,蒋细旺,等.药用植物生物技术.合肥:中国科学技术大学出版社,2005.1~3
[8] 李文安,余叔文.植物生理与分子生物学.北京:科学技术出版社,1992,37
[9] 翟中和,王喜忠,丁明孝,等.细胞生物学.北京:高等教育出版社,2000
[10] 谷瑞升,蒋湘宁,郭仲琛,等.植物离体培养中器官发生调控机制研究进展.植物学通报.1999,16(3):238~244
[11] 李明军,薛建平,陈明霞,等.不同因子对山药愈伤组织诱导的影响.广西植物,2000,20(2):156~160
[12] 黄潞,卫志明.不同基因型玉米的再生能力和胚性与非胚性愈伤组织 DNA 的差异.植物生理学报.1999,25(4):332~338
[13] Piagnani C,Bassi D.In vivo and in vitro propagagation of Sorbus spp.from juvenile material.Italus Hortus.2000,7(5):3~7
[14] 黄学森,焦定量,那丽,等.西瓜子叶离体培养获得再生植株.中国西瓜甜瓜.1994,(3):15~16
[15] Preece JE . A century of progress with vegetative plant propagation.HortScience.2003,38:1015~1025
[16] 罗凤霞,徐桂华,金丽丽,等.新铁炮百合微繁的研究.沈阳农业大学学报,2000,31(3):254~257
[17] 王刚,杜捷,李桂英,等.看周百合和野百合组织培养及快速繁殖研究.西北师范大学学报(自然科学版),2002,38(1):69~71
[18] 刘敏.花卉组织培养与工业化生产.北京:地质出版社.2002:44
[19] Wagner J,Kafka I.Effect of medium composition on in vitro germination of embryos of Fraxinus excelsior at different stages of development.Plant Physiology.1995,146:566~568
[20] 兰素缺,李光威,权书月,等.碳源组分及浓度对小麦花药培养和游离小孢子培养的影响.华北农学报.2002,17(增刊):92~97
[21] 丁世萍,严菊强,季道藩,等.糖类在植物组织培养中的效应.植物学报.1998,15(6):42~46
[22] 王爱民,刘文,傅中滇,等.不同碳源对红边朱蕉组培苗生长的影响.徐州示范大学学报.2003,21(3):76~78
[23] Hadrami EL,Dauac J.Effcets of growth regulators on polyamine content and peroxidase activity in Hebea brasilienses callus.Ann,Bot,1992,69:323~325
[24] 詹亚光,样传平.白桦愈伤组织的高效诱导和不定芽分化.植物生理学通讯.2002,38(2):117~114
[25] Martinelli L,Gianazza E.Biochemical changes during regeneration of sunflower (Helianthus annuus L.).Electrophoresis.1996,17(1):191~197
[26] Michalczuk L,Ribnicky DM,Cooke TJ,et al.Regulation of indole-3-aceticacid biosynthetic pathway in carrot cell cultures.Plant Physiol.1992,100:1346~1353
[27] Leshem B,Ronen R,Soudry E,et al.Cytokinin at a large range of concentration determines rates of polypeptide metabolism and regeneratiom in cultere melon cotyledons.Plant Physiol.1994,143:330~336
[28] Kozai T,Tanaka K,Jeong BR,et al.Effect of relative humidity in the culture vessel on the growth and shoot elongation of potato plantlets in vitro.Journal of the Japanese Society for Horticultural Science.1993,62(2):413~417
[29] Reed BM,Mentzer J,Tanpraser P,et al.Internal Bacterial contamination of micropropagated hazelnut:indentification and antibiotic treatment.Plant Cell.Tissue and organ Culture.1998,(52):53~55
[30] Kritzinger EM,Vanbvuuren RJ,Woodward B,et al.Elimination of external and internal contamination in rhizomes of Zantedeschia aethiopica with commercial fungicides and antibiotics.Plant Cell.Tissue and Organ Culture.1998,(52):61~65
[31] 高国训.植物组织培养中的褐变问题.植物生理学通讯.1999,35(6):501~506
[32] 刘兰芳."薄壳香"核桃组织培养中的褐化及防止措施研究.园艺学报.2002,29(2):171~172
[33] 姚洪军,罗晓芳.植物组织培养外植体河边的研究进展.北京林业大学学报.1999,21(3):78~84
[34] Dan H,Meredith CP.The influence of explant origin on tissue browning and shoot production in shoot tip cultures of grapevine.Journal of the American Society for Horicultural Science.1986,25:456~466
[35] 程家胜,史永中,张志云,等.苹果组织培养中的玻璃苗问题.植物生理学通讯.1991,(1):33~35
[36] 郝瑞庆,杨广东.大白菜试管苗玻璃化发生机制初探.中国农学通报.2002,18(3):45~47
[37] 丁运华,王鸿博.3 种应诉对梅菜试管苗玻璃化的影响.热带农业科学.2003,23(23):5~9
[38] 于树宏,李玲.野葛的组织培养和植株再生.植物资源与环境.1998,8(1):63~64
[39] 施和平.三裂叶野葛茎段和叶柄培养再生植株.中草药.2000,31(7):550~552
[40] 周堂英,李惠波,向素琼,等.粉葛组织培养及同源四倍体诱导.2005,36(8):1230~1233
[41] 吴丽芳,张素芳,蒋亚莲,等.粉葛的离体培养和无糖生根.2005,41(5):646~647
[42] 夏英武.作物诱变育种.北京:中国农业出版社.1997.84~91
[43] 刘军,核科学技术在农业领域的应用.核科学百年讲座.2001
[44] 徐冠仁.核农学导论.北京:原子能出版社.1995
[45] Charbaji TN.Effect of low doses of gamma irradiation on in vitro growth ofgrapevine.Plant Cell,Tissue and Organ Culture,1999,57:129~132
[46] 吴关庭等.小麦辐射育种研究进展.核农学通报.1993.14:189~191
[47] 韩月澎,陈秀兰,何震天,等.大麦辐射育种研究进展.大麦科学.2000.3:11~13
[48] 杨爱珠.玉米辐射诱变的研究和应用.核农学通报.1995,16:158~159
[49] 万贤国.辐射诱变在创造种质资源上的应用作物研究.1994.8:1~4
[50] Ahloowalia BS,Maluszynski M:Induced mutations-A new paradigm in plant breeding Euphytica,2001,118:167~173
[51] 毛炎麟.辐射的生物学效应.植物诱变育种学北京:中国农业出版社.1996.79~100
[52] 杨期和,尹小娟,叶万辉,等.硬实种子休眠的机制和解除方法.植物学通报.2006.23(1):108~118
[53] 孙仲序,王玉军,刘静,等.植物组织快繁滤纸纸桥生根新技术[J].山东农业大学学报(自然科学版),2002,33(3):257~263
[54] Edwin BH.Non-axenic plant tissue culture:possibilities and opportunities [J].Acta Horticulture,1990,280:1990
[55] Flowler MW,Wastson R.In proc.5th Ind.Cong [J].Plant Tissue and Cell Culture.1982,Tissue Culture pp 225
[56] Ewing EE.The role of hormones in potato(Solanum tuberosum L.)tuberization.In:Davies P J ed.Plant hormones physiology,biochemistry and molecular biology.(2nd ed).Dordrecht:Kluwer Academic Pub,1995,698~724
[57] Khuri S,Moorby J.Investigations into the role of sucrose in potato cv.Estima microtuber production in vitro.Ann of Bot,1995,75:295~303
[58] 连勇,邹颖,杨宏福,等.马铃薯试管薯发育机理研究-温度对试管薯形成的影响.马铃薯杂志,1996a,10:133~137
[59] 沈清景,叶贻勋,凌永胜,等.马铃薯试管薯诱导因素研究,福建农业学报,2001,16(1):54~56
[60] 柳俊,谢从华.马铃薯块茎发育机理及其基因表达.植物学通报,2001,18(5):531~539
[61] 蒙美莲,刘梦芸,门福义,等.赤霉素和脱落酸对马铃薯块茎形成的影响.马铃薯杂志,1994,8 (3):134~137
[62] Stefano P.Mutation induction and tissue culture in improving fruits.Plant Cell,Tissue and Organ Culture,2001,64:185~2101
[63] 徐华军,贺源辉,陈秀芳,等.60Coγ 射线对双低甘蓝型油菜的辐射效应.核农学报.1992.6:199~206
[64] 宋恒,王长泉,巩向忠,等.γ 射线辐照杜鹃试管苗诱发突变体的研究.核农学报.2003.17:347~349
[65] 李林光,孙清荣,贺桂香,等.苹果离体叶片 60Coγ 辐射对不定稍再生及芽生长的影响.山东农业科学.2004:36~38
[66] 翁伯琦,徐国忠,郑向丽,等.60Coγ 射线辐照圆叶决明子对其生物学特性的影响.核农学报.2004.15:197~200
[67] 宋淑敏,田玉杰,姬研茹,等. γ 射线辐射亚麻花药的研究初报.中国麻业.2004.14:162~176
[68] Rama N,Swamy UT,Praveen M.Induction of streptomycin-resistant plantlets insolanum surattense through in vitro mutagenesis.Plant cell,Tissue and Organ Culture,2005,80:201~207
[69] Amer IM , ElRassas HN . Utilization of gamma irradiation in barley breeding.Annals of Agricultural Science (Cairo),1994,39:257~268
[70] Datta SK.Chrysanthemum cultivars evolved by induced mutation at National Botanical Research Institute,Lucknow.The Chrysanthemum,44:72~75
[71] Forster BP,Pakniyat H,Macaulay M,et al.Variation in the leaf sodium content of Hordeum vulgare (barley) cultivar Maythorpe and its derived mutant cv . Golden Promise.Heredity,73:249~253
[72] Ahloowalia BS.In-vitro techniques and mutagenesis for the improvement of vegetatively propagated plant.In:Jain M S,Brar D S,Ahloowalia B S(Eds.).Somaclonal Variation and Induced Mutations in Crop Improvement.293~309.Kluwer Academic Publishers,Dordrecht,NL.1998
[73] 王文恩,张俊卫,包满珠,等.60Coγ 对野牛草干种子的刺激生长效应.核农学报.2005.19:191~194.
[74] 杨期和,尹小娟,叶万辉,等.硬实种子休眠的机制和解除方法.植物学通报.2006.23(1):108~118
[75] 孙仲序,王玉军,刘静,等.植物组织快繁滤纸纸桥生根新技术[J].山东农业大学学报(自然科学版),2002,33(3):257~263