光质对铁皮石斛体细胞胚胎发生的影响
|
摘要
光是植物生长发育最重要的环境因子之一,现已广泛应用于植物组织培养过程中的形态发生、生长控制、光形态建成及其各种反应机理的研究中,大量研究结果表明,各种光质对植物体细胞胚胎发育中的形态发生、器官分化、生化指标等都有一定程度的影响。本实验以铁皮石斛原球茎诱导得到的胚性愈伤组织为原始材料,经过红光NO.1、红光NO.2、绿光、蓝光、白光、黑暗6种光质处理,在形态学水平上观察体胚发育过程中一些形态特征的变化,并对过氧化物酶(POD)、酯酶(EST)、细胞色素氧化酶(COD)的同工酶谱进行了分析。
对各种光质下铁皮石斛体胚的形态学研究发现,在42d的培养期中,光质对铁皮石斛体胚生长量有不同程度的促进作用:蓝光＞白光＞红光NO.1＞红光NO.2＞绿光＞黑暗;而不同光质下每个生长时期内每克鲜重的增长率顺序为:白光＞蓝光＞红光NO.1＞绿光＞红光NO.2＞黑暗。对铁皮石斛体胚颜色变化的观察发现,绿光和白光能够使组织块更快地变绿,蓝光次之。
本实验对6种光质条件下POD、EST、COD的同工酶谱的酶带数、酶带强度、酶带Rf值进行了分析,并且找到了每种光质都共有的酶带:POD同工酶带Rf_(P16)、Rf_(P20);EST同工酶带Rf_(E7)、Rf_(E11);COD同工酶带Rf_(C13)、Rf_(C15)。此外也发现了某种光质特有的酶带14条:红光NO.1下3条;绿光和蓝光下各1条:白光下3条;黑暗下6条。将酶带的特征与形态学特征结合,我们可以通过同工酶的指示初步的了解铁皮石斛体胚在不同光质下的发育过程。
Light is one of the most important environmental factors for plant growth and development.As a physical factor,it has been widely used in plant tissue cultures for morphogenesis,growth control,photomorphogenesis and its reaction mechanism.Results of many researches have shown that the light has a great effect on morphogenesis,organ differentiation and biochemical indicators in plant callus growth.In this study,the embryonic calli was used as the original material,which were induced by protocorm-like bodies(PLBs)of Dendrobium candidum Wall ex Lind.6 kinds of light quality:red No.1,red No.2,green,blue, white and dark were applied in the culture.During somatic embryogenesis, morphogenesis and isozymes of peroxidase(POD),esterase(EST)and cytochrome oxidase(COD)were analyzed.
The somatic embryos of Dendrobium candidum were observed in morphology and it was found that different lights could take different extents effects on somatic embryos during 42 days of culture:the Blue＞White＞red No.1＞Red No.2＞Green＞dark;and in each growth period the order of the growth rate of fresh weight per gram was:White＞Blue＞red No.1＞Green＞red No.2＞Dark.The color of somatic embryos in Dendrobium candidum changed from yellow to green.Under white light and green light,the speed of greening was faster than under blue.
In this study,through analysis of POD,EST,COD isozyme zymogram,it was found that the light was related with gene expression of the isozymes.And the common isozyme band was found for each kind of the light:POD isozyme with Rf_(P1)Rf_(P2),Rf_(P3),Rf_(P1)and Rf_(P2)were in each of the eight periods;EST isozyme with Rf_(E1),Rf_(E2),and both of them were in each of the eight periods;COD isozyme with Rf_(C1),Rf_(C2),Rf_(C3),Rf_(C4),Rf_(C5),they were in great difference in each period.In addition,we also found 14 specific bands:red No.1 light with three; green light and blue light each with one;white light with three;darkness with six. To combine with the morphological characteristics and the isozyme zymogram characteristics,we could preliminary understand the development process of the somatic embryos in Dendrobium candidum through the indication of isozyme zymogram.
对各种光质下铁皮石斛体胚的形态学研究发现,在42d的培养期中,光质对铁皮石斛体胚生长量有不同程度的促进作用:蓝光＞白光＞红光NO.1＞红光NO.2＞绿光＞黑暗;而不同光质下每个生长时期内每克鲜重的增长率顺序为:白光＞蓝光＞红光NO.1＞绿光＞红光NO.2＞黑暗。对铁皮石斛体胚颜色变化的观察发现,绿光和白光能够使组织块更快地变绿,蓝光次之。
本实验对6种光质条件下POD、EST、COD的同工酶谱的酶带数、酶带强度、酶带Rf值进行了分析,并且找到了每种光质都共有的酶带:POD同工酶带Rf_(P16)、Rf_(P20);EST同工酶带Rf_(E7)、Rf_(E11);COD同工酶带Rf_(C13)、Rf_(C15)。此外也发现了某种光质特有的酶带14条:红光NO.1下3条;绿光和蓝光下各1条:白光下3条;黑暗下6条。将酶带的特征与形态学特征结合,我们可以通过同工酶的指示初步的了解铁皮石斛体胚在不同光质下的发育过程。
Light is one of the most important environmental factors for plant growth and development.As a physical factor,it has been widely used in plant tissue cultures for morphogenesis,growth control,photomorphogenesis and its reaction mechanism.Results of many researches have shown that the light has a great effect on morphogenesis,organ differentiation and biochemical indicators in plant callus growth.In this study,the embryonic calli was used as the original material,which were induced by protocorm-like bodies(PLBs)of Dendrobium candidum Wall ex Lind.6 kinds of light quality:red No.1,red No.2,green,blue, white and dark were applied in the culture.During somatic embryogenesis, morphogenesis and isozymes of peroxidase(POD),esterase(EST)and cytochrome oxidase(COD)were analyzed.
The somatic embryos of Dendrobium candidum were observed in morphology and it was found that different lights could take different extents effects on somatic embryos during 42 days of culture:the Blue＞White＞red No.1＞Red No.2＞Green＞dark;and in each growth period the order of the growth rate of fresh weight per gram was:White＞Blue＞red No.1＞Green＞red No.2＞Dark.The color of somatic embryos in Dendrobium candidum changed from yellow to green.Under white light and green light,the speed of greening was faster than under blue.
In this study,through analysis of POD,EST,COD isozyme zymogram,it was found that the light was related with gene expression of the isozymes.And the common isozyme band was found for each kind of the light:POD isozyme with Rf_(P1)Rf_(P2),Rf_(P3),Rf_(P1)and Rf_(P2)were in each of the eight periods;EST isozyme with Rf_(E1),Rf_(E2),and both of them were in each of the eight periods;COD isozyme with Rf_(C1),Rf_(C2),Rf_(C3),Rf_(C4),Rf_(C5),they were in great difference in each period.In addition,we also found 14 specific bands:red No.1 light with three; green light and blue light each with one;white light with three;darkness with six. To combine with the morphological characteristics and the isozyme zymogram characteristics,we could preliminary understand the development process of the somatic embryos in Dendrobium candidum through the indication of isozyme zymogram.
引文
[1]赵鹏,冯佛生,王万军.铁皮石斛愈伤组织的诱导及再生植株.四川省遗传学会第七届代表大会暨学术讨论会.2004
[2]李淑兰.高等植物的体细胞胚胎发生研究.山东省农业管理干部学院学报.2001 150-151
[3]Gupta PK,Pullman GS.Method for reproducing coniferous plants by somatic embryogenesis using adscisic acid and somatic potential varition[J].U S Patent,1991;5:36.
[4]汤浩茹,王永清,任正隆.果树的体细胞胚发生[J].四川农业大学学报,1999:17(1):69-79
[5]杨金玲,桂耀林,杨映根等.白杆体细胞胚胎发生及其植株再生[J].植物学报,1999:39:315-321.
[6]Biahoua A,Bonneau L.Control of in vitro somatic embryogenesis of the spindle tree(Eu- nymus europaeus L.)by the sugar type and the osmotic potential of the culture medium[J].Plant Cell Reports,1999;185-190.
[7]严菊强.糖源对植物组织培养的影响.中国植物生理学会第二届青年植物生理工作者会议论文摘要汇编,1994;24-26.
[8]达克东,张松,李雅芝等.苹果离体叶片培养直接体细胞胚胎发生研究[J].园艺学报,1996;23(3):241-245.
[9]Harsh Pal Bais,Ravishankar GA.Role of polyamines in the ontogeny of plants and their bioteclmological applications[J].Plant Cell Tissue and Organ Culture,2002;69:1-34.
[10]邢登辉,赵云云,黄承芬.皇冠草体细胞胚发生及其体胚发生过程中内源激素的变化[J].生物工程学报,1999,15(1):98-103.
[11]肖关丽,杨清辉,李富生,等.甘蔗愈伤组织分化绿苗内源激素变化规律研究 [J].西南农业大学学报,2002,24(4):337-338.
[12]郭子彪,盖钧镒.内源激素IAA、ABA对大豆萌发子叶胚性愈伤组织诱导及其分化的调控[J].大豆科学,1997,16(3):194-198
[13]周俊彦,郭扶兴.细胞分裂素类物质在植物体细胞胚发生中作用[J].植物生理学通讯,1996,32(4):247-253.
[14]崔凯荣,邢更生,周公克,等.植物激素对体细胞胚胎发生的诱导与调节[J].遗传,2000,22(5):349-354.
[15]崔凯荣,邢更生,刘新民,等.细胞信号转导与植物体细胞胚发生[J].生命科学,2002,14(3):171-175
[16]刘华英,萧浪涛,何长征.植物体细胞胚发生与内源激素的关系研究进展[J].湖南农业大学学报,2002,28(4):350-354.
[17]郏艳红,熊庆娥.植物体细胞胚发生的研究进展[J].四川农业大学学报,2003,21(1):59-63.
[18]韩碧文,刘淑兰.植物离体体细胞胚胎发生[J].植物生理学通讯,1988,(10):9-15.
[19]周俊彦.植物体细胞在组织培养中产生的胚状体Ⅱ.影响植物胚状体发生和发育的因素[J].植物生理学报,1982,8(1):91-99
[20]汤浩茹,王永清,任正隆.核桃体细胞胚发生与转基冈研究进展[J].林业科学,2000,36(3):102-110.
[21]佟曦然,张琪,高微微,等.影响人参种胚体细胞胚胎发生的几个因素[J].植物学通报,1991,8(增刊):43-47.
[22]贺红,潘瑞炽,韩美丽,等.红江橙体胚发生及影响因素的研究[J].广西植物,1998,18(4):343-346.
[23]杨金玲,赵德修,桂耀林,等.水母雪莲体细胞胚胎发生及其植株再生[J].西北植物学报,2001,21(2):252-256.
[24]Jain SM,Gupta PK,Newton RJ.Somatic embryogenesis in woody plants[M]. Dordrecht:Kluwer Acdemic Publishers,1995;17-143.
[25]Carimi F,De Pasquale F,Crescimanno FG.Somatic embryogenesis and plant regeneration from pistil thin cell layers of Citrus[J].Plant Cell Reports,1999;18:935-940.
[26]唐巍,欧阳藩,郭仲琛。火炬松成熟合子胚培养直接体细胞胚胎发生和植株再生[J],应用与环境生物学报,1998,4(2):103-106
[27]Hofer M,Touraev A.Heberle-Bors E.Induction of embryogenesis from isolated apple microspores[J].Plant Cell Report,1999;18:1012-1017.
[28]周丽侬,邝哲师,马雪筠,等.荔枝幼胚培养及体细胞胚胎发生研究初报[J].广东农业科学,1993,(5):14-15.
[29]周丽侬,曹静,邝哲师,等.园艺植物体胚发生及植株再生技术研究[J].热带作物学报,1998,19(2):15-19.
[30]邝哲师,周丽侬,马雪筠,等.香蕉和荔枝体胚发生两种不同途径的研究[J].农业生物技术学报,1998,6(3):210-222.
[31]曹静,周丽侬,邝哲师,等.白鹤芋花序体细胞胚胎发生及植株再生的研究[J].农业生物技术学报,1995,3(3):81-85.
[32]黄学林,李筱菊.高等植物组织离体培养的形态建成及其调控[M].北京:科学出版社,1995.
[33]Liao YK,Amerson HV.Slash pine(pinus elliottii Engelm)somatic embryogenesis.I.Inittiation of embryogenic cultures from immature zygotic embryos[J].New For,1995;10:145-163.
[34]杨利平,程井辰。植物体细胞胚胎发生中的生理生化研究进展[J]。植物学通报,1991,8(2):1-8
[35]郭祖宝,王成增,孙培良。植物体细胞胚胎发生中的生理生化变化[J]。安徽农学通报,2007,13(14):52-53
[36]周燕,高述民等。胡萝卜体细胞胚胎发生中的细胞组织化学和蛋白质组分变 化[J]。植物生理学通讯,2004,40(2):181-183
[37]高述民,陆帼一,杜慧芳等。大蒜体细胞胚发育分化中特异蛋白质和某些生理生化变化[J]。植物生理学通讯,2001,37(3):207-23
[38]贺晓,慈忠玲等。紫花苜蓿体细胞胚胎发生及其细胞组织学观察[J]。内蒙古林学院学报(自然科学版),1999,21(2):44-46
[39]徐竹绮,王彤宁,何美瑛,等.水稻离体体细胞胚与合子胚的某此生化特性的比较[J].植物生理学报,1995,21(1):95-102.
[40]Kamada H.Harada H.Changes in endogenous amino acid compositions during somatic embryogenesis in Daucus carota L[J].Plant & Cell Physiol,1984,25:27-28
[41]杨和平,程井辰.马唐胚性和非胚性愈伤组织生理差异的初步研究[J]。植物生理学通讯,1991,27:337-34
[42]王亚馥,崔凯荣等。小麦体细胞胚发生的蛋白质组分和过氧化物酶同工酶的变化[J]。兰州大学学报(自然科学版),1993,29(3):189-193
[43]郑晓峰,黄百渠,几种植物体细胞胚胎发生标记蛋白的研究[J]。植物学报,1994,36(3):175-180
[44]Stirn S Jacobsen H.J.Marker proteins for embryogenic differentiation patterns in pea callus.Plant Cell,1987(6):50-54
[45]Chen L.J.Lathe D.S.Analysis of proteins from embryogenic and non-embryogenic rice(Oryza satire L·)calli·Plant Sci·1987,48:181-188
[46]崔凯荣,邢更生,周功克,等.体细胞胚发生的生化基础[J].生命科学,2001,13(1):28-33.
[47]戴若兰,张玮,邢更生,等.枸杞体细胞胚发生中DNA代谢动态的立体计量[J].植物学通报,2000,17(1):80-83
[48]Fujimura T,Komamine A,Matsumoto H.Aspects of DNA,RNA and protein synthesis during somatic embryogenesis in a carrot cell suspension culture. Physiol Plant,1980(49):255-260
[49]Ashibara H Z.Pflartzen et al.,1981,physil.,104:129-137
[50]Seugupta,Cand V.RagLavan,K.Exp.Bot.,1980(31):217-258
[51]Sevgupts,Cand V.Raghavan,K.Exp.Bot.,1980(31):259-268
[52]徐竹绮.2,4-D浓度对胡萝卜体细胞胚胎发生及其同工酶的影响[J].植物生理学报,1984,10(4):373-379
[53]Chibbar R.N et al.[J].Plant Physicl.,1988,133:367-370
[54]余沛涛,薛应龙.植物苯丙氨酸解氨酶(PAL)在细胞分化中的作用[J].植物生理学报,1987,13(1):14-19
[55]李国珍,秦明波,孙成华,等.青蒿体细胞胚胎发生及遗传转化的研究[J].植物学通报 1992,9(增刊)97-98
[56]崔凯荣,刑更生,王亚馥,等.枸杞组织培养中抗氧化酶活性与体细胞胚发生相关性的研究[J].兰州大学学报,1998,34(3):93-99.
[57]潘有福,王星,孟玉玲,等.秦艽体细胞胚不同发育阶段几种同工酶的特性[J].西北植物学报,1994,15(5):6-10
[58]陈耀锋,贺普超,廖祥和,等.葡萄脯氨酸累积变异系CAT和SOD活性研究[J].西北农业大学学报.1998,26(1):36-40.
[59]陈善春,张进仁,吴安仁,等.柳橙胚性愈伤组织耐盐性研究[J].西南农业大学学报,1987,9(4):463-468.
[60]林定波,颜秋生,沈德绪.柑橘抗寒细胞变异的获得及其抗性遗传稳定性的研究[J].植物学报,1999,41(2):136-141.
[61]胡含,王恒立.植物细胞工程与育种[M].北京:北京工业大学出版社,1990.
[62]郭梅.甘薯胚性恳浮细胞培养中的体细胞无性系变异分析[D].北京:中国农业大学,2005
[63]De capite L,Action of light and temperature on growth of plant tissue culture in vitro[J].Am.J.Bot.,1955,42:869-873.
[64]Seibert M,Wetherbee P J,Job D D,The effects of light intensity and spectral quality on growth and shoot initiation in tobacco callus[J].Plant Physiol.,1975,56:130-139.
[65]宋云,唐崇钦,匡廷云,等.不同光质下生长的满江红叶绿体和共生者满江红鱼腥藻的吸收光谱和荧光光谱的研究[J].植物生理学报,1983,9(1):69-76
[66]陶汉之,王新长.茶树光合作用与光质的关系[J].植物生理学通讯,1989,25(1):19-23
[67]范慈惠.光质对植物愈伤组织的诱导和植株形成的影响[J].植物生理学通讯,1981,17(1):25-33
[68]倪德祥,易莹,张丕方,等.光质对凹叶景天愈伤组织和不定芽形成的效应[J].上海农业学报,1988,4(1):43-48
[69]王维荣,王咏冬,欧阳光察,等.光质对黄瓜及番茄愈伤组织培养中分化和有关酶的影响[J].植物生理学报,1991,17(2):118-124.
[70]许桂芳,董诚明,周吉源,等.不同光质对华黄芪愈伤组织诱导、增殖及器官分化的效应[J].华中师范大学学报(自然科学版),1994,28(4):533-537.
[71]熊丽,周吉源,殷荣华.光质对石刁柏愈伤组织培养中生长和过氧化物酶的影响[J].武汉植物学研究,1995,13(3):253-257.
[72]阎秀峰,王洋,尚辛亥,等.光强和光质对野外栽培高山红景天生物量和红景天甙含量的影响[J].生态学报,2004,24(4):674-679
[73]赵德修,李茂寅,邢建民,等.光质、光强和光期对水母雪莲愈伤组织生长和黄酮生物合成的影响[J].植物生理学报,1999,25(2):127-13
[74]沈红香,沈漫,程继鸿,等.不同光质补光处理对郁金香生长和开花的影响[J].北京农学院学报,2007,22(1):16-18
[75]何小弟,裴建文,黄春华.菊花组织培养中的光质效应研究[J].江苏林业科技,2002,29(6):26-28
[76]周波,李玉花.植物的光敏色素与光信号转导[J].植物生理学通讯,2006, 42(1):134-140
[77]徐根娣,李瑶.光质和碳素对旱芹体细胞胚发生的影响[J].浙江师大学报,1995,18(2):57-60
[78]洪森荣,尹明华.光质对野葛叶片愈伤组织的诱导及其可溶性蛋白含量的影响[J].安徽农业科学,2007,35(9):2560-2563
[79]毛学文,陈荃.不同光质对毛地黄愈伤组织诱导和增值的效应[J].植物学通报,1997,14(1):55-56
[80]张君诚.银杏愈伤组织诱导与增殖的效应研究[J].重庆师范学院学报(自然科学版),1999,16(3):41-48
[81]梁钾贤,陈彪.光质对甘蔗愈伤组织组织分化出苗的影响[J].中国糖料,2006(3):9-11.
[82]Rey M,Diaz S,Rodriguez R.Exogenous polyamines imp rove rooting of hazelmicroshoots[J].Plant Cell Tissue Ore.Culture,1994,36(3):303-308
[83]倪德祥,陈刚,张丕方,等.锦葵愈伤组织培养中光质对其过氧化物同工酶和发根的效应[J].复旦学报(自然科学版),1986,25(2):157-162.
[84]倪德祥,王维荣,张丕方,等.菊花器Ⅱ光质和组合激素对菊花茎段培养中生长发育的影响[J].西北植物学报,1988,8(1):16-27
[85]李韶山,潘瑞炽.植物的蓝光效应[J].植物生理学通讯,1993,29(4):248-252
[86]王学奎,李合生,刘武定.光对小麦叶片谷氨酰胺合成酶调节机理初探[J].华中农业大学学报,2000,19(2):102-105
[87]王莉,史玲玲,刘玉军.不同光质对长鞭红景天悬浮细胞生长及苯丙氦酸解氨酶活性的影响[J].林业科学,2007,43(6):52-56
[88]Anna B,Alicjia K.Effect of light quality on somatic embryogenesis in Hyacinthus orientalis L.'Delft's blue'[J].Biological Bulletin of Poznan,2001,38(1):103-107
[89]梁学芬,蚁伟南,颜梓兴,等.不同光质的辅助光对香蕉组培苗的波长的影响[J].中国南方果树,2001,30(4):34
[90]Kasemir H.,Obdorfer U.,Mohr H.A twofold action of phytochrome in controlling chlorophyll a accumulation[J].Photochem,Photobiol,1973,18:481-486
[91]倪德祥,王蓓,王维荣,等.不同波长的光对佛甲草离体茎段培养过程中生长发育的影响[J].云南植物研究,1988,10(1):113-116
[92]Singla N K,Jain V,Jain S,et al.Activities of glycolytic enzymes in leaves and roots of contrasting cultivars of sorghum during flooding[J].Biologia Plantarum,2003,47(4):555-560
[93]Ferriol M,Pico B,Nuez F.Genetic diversity of a germplasm collection of cucurbita pepo using SRAP and AFLP markers[J].Heor Appl Genet,2003,107(2):271-282.
[94]倪德祥,冯文煦,张丕方,等.光质对一品红叶离体培养中形态发生及生化变化的影响[J].广西植物,1987,7(4):339-345
[95]倪德祥,易莹,张丕方,等.光质对凹叶景天愈伤组织和不定芽形成的效应[J].上海农业学报,1988,4(1):43-48
[96]Chen W,Cun SX.In vitro rapid propagation of stems of Dendrobium candidum.Plant Physiol.Commun.2002,(38):145.
[97]Shiau YJ,Nalawade SM,Hsia CN,Mulabacal V,Tsay HS.In Vitro propagation of the Chinese medicinal plant,Dendrobium Candidum Wall.Ex Lindl.,from axenic nodal segments.In Vitro Cell.Dev.Biol-Plant.2005,(41):666-670.
[98]He JB,Zheng CZ,Wang SL.Multiplication of protocorm of Dendrobium candidum.Acta.Botanica Yunnanica.1982,(4):211-212.
[99]Ye XL,Cheng SJ,Wang F,Qian NF.Morphology of immature seeds and development in vitro of Dendrobium candidum.Acta.Botanica Yunnanica.1988,(10):285-290.
[100]Zhang QX,Fang YM.Tissue culture and in vitro seedling and protocorm-like body examination of Dendrobium candidum.Acta.Botanica Boreali-Occidentalia Sinica.2005,(25):1761-1765.
[101]ZhaoP,Wang W,Feng FS,Wu F,Yang ZQ,Wang WJ.High-frequency shoot regeneration through transverse thin cell layer culture in Dendrobium Candidum Wall Ex Lind,Plant Cell,Tissue and Organ Culture,2007(90):131-139
[102]李阳春,谢可军.早熟禾属10种植物酯酶同工酶分析[J].草地学报,2003,11(3):214-219
[103]Hazard L,Ghesouiere M.Evidence from the use of isozyme markers of competition in swards between short leaved and long-leaved perennial ryegrass[J].Grass and Forage Science,1995,50:241-248
[104]Liu TL,Jiang XW,Wang LS.The analysis of special proteins and isoenzymes in globular embryogenesis of alfalfa in tissue culture.Acta.Botanica Boreali-Occidentalia Sinica.2002,(3):625-628.
[105]Everett NP,Wach MJ,Ashworth DJ.Biological marders of embryogenesis in tissue cultures of the mazie inbred in B73.Plant Sci.1985,(41):133-140
[106]Pan YF,Wang X,Meng YL,Wang LS.Characterization of several isoenzymes from embryogenic calli and somatic embryos at different developmental stages Gentiana Crassicaulis.Acta.Botanica Boreali-Occidentalia Sinica.1994,(15):6-10.
[2]李淑兰.高等植物的体细胞胚胎发生研究.山东省农业管理干部学院学报.2001 150-151
[3]Gupta PK,Pullman GS.Method for reproducing coniferous plants by somatic embryogenesis using adscisic acid and somatic potential varition[J].U S Patent,1991;5:36.
[4]汤浩茹,王永清,任正隆.果树的体细胞胚发生[J].四川农业大学学报,1999:17(1):69-79
[5]杨金玲,桂耀林,杨映根等.白杆体细胞胚胎发生及其植株再生[J].植物学报,1999:39:315-321.
[6]Biahoua A,Bonneau L.Control of in vitro somatic embryogenesis of the spindle tree(Eu- nymus europaeus L.)by the sugar type and the osmotic potential of the culture medium[J].Plant Cell Reports,1999;185-190.
[7]严菊强.糖源对植物组织培养的影响.中国植物生理学会第二届青年植物生理工作者会议论文摘要汇编,1994;24-26.
[8]达克东,张松,李雅芝等.苹果离体叶片培养直接体细胞胚胎发生研究[J].园艺学报,1996;23(3):241-245.
[9]Harsh Pal Bais,Ravishankar GA.Role of polyamines in the ontogeny of plants and their bioteclmological applications[J].Plant Cell Tissue and Organ Culture,2002;69:1-34.
[10]邢登辉,赵云云,黄承芬.皇冠草体细胞胚发生及其体胚发生过程中内源激素的变化[J].生物工程学报,1999,15(1):98-103.
[11]肖关丽,杨清辉,李富生,等.甘蔗愈伤组织分化绿苗内源激素变化规律研究 [J].西南农业大学学报,2002,24(4):337-338.
[12]郭子彪,盖钧镒.内源激素IAA、ABA对大豆萌发子叶胚性愈伤组织诱导及其分化的调控[J].大豆科学,1997,16(3):194-198
[13]周俊彦,郭扶兴.细胞分裂素类物质在植物体细胞胚发生中作用[J].植物生理学通讯,1996,32(4):247-253.
[14]崔凯荣,邢更生,周公克,等.植物激素对体细胞胚胎发生的诱导与调节[J].遗传,2000,22(5):349-354.
[15]崔凯荣,邢更生,刘新民,等.细胞信号转导与植物体细胞胚发生[J].生命科学,2002,14(3):171-175
[16]刘华英,萧浪涛,何长征.植物体细胞胚发生与内源激素的关系研究进展[J].湖南农业大学学报,2002,28(4):350-354.
[17]郏艳红,熊庆娥.植物体细胞胚发生的研究进展[J].四川农业大学学报,2003,21(1):59-63.
[18]韩碧文,刘淑兰.植物离体体细胞胚胎发生[J].植物生理学通讯,1988,(10):9-15.
[19]周俊彦.植物体细胞在组织培养中产生的胚状体Ⅱ.影响植物胚状体发生和发育的因素[J].植物生理学报,1982,8(1):91-99
[20]汤浩茹,王永清,任正隆.核桃体细胞胚发生与转基冈研究进展[J].林业科学,2000,36(3):102-110.
[21]佟曦然,张琪,高微微,等.影响人参种胚体细胞胚胎发生的几个因素[J].植物学通报,1991,8(增刊):43-47.
[22]贺红,潘瑞炽,韩美丽,等.红江橙体胚发生及影响因素的研究[J].广西植物,1998,18(4):343-346.
[23]杨金玲,赵德修,桂耀林,等.水母雪莲体细胞胚胎发生及其植株再生[J].西北植物学报,2001,21(2):252-256.
[24]Jain SM,Gupta PK,Newton RJ.Somatic embryogenesis in woody plants[M]. Dordrecht:Kluwer Acdemic Publishers,1995;17-143.
[25]Carimi F,De Pasquale F,Crescimanno FG.Somatic embryogenesis and plant regeneration from pistil thin cell layers of Citrus[J].Plant Cell Reports,1999;18:935-940.
[26]唐巍,欧阳藩,郭仲琛。火炬松成熟合子胚培养直接体细胞胚胎发生和植株再生[J],应用与环境生物学报,1998,4(2):103-106
[27]Hofer M,Touraev A.Heberle-Bors E.Induction of embryogenesis from isolated apple microspores[J].Plant Cell Report,1999;18:1012-1017.
[28]周丽侬,邝哲师,马雪筠,等.荔枝幼胚培养及体细胞胚胎发生研究初报[J].广东农业科学,1993,(5):14-15.
[29]周丽侬,曹静,邝哲师,等.园艺植物体胚发生及植株再生技术研究[J].热带作物学报,1998,19(2):15-19.
[30]邝哲师,周丽侬,马雪筠,等.香蕉和荔枝体胚发生两种不同途径的研究[J].农业生物技术学报,1998,6(3):210-222.
[31]曹静,周丽侬,邝哲师,等.白鹤芋花序体细胞胚胎发生及植株再生的研究[J].农业生物技术学报,1995,3(3):81-85.
[32]黄学林,李筱菊.高等植物组织离体培养的形态建成及其调控[M].北京:科学出版社,1995.
[33]Liao YK,Amerson HV.Slash pine(pinus elliottii Engelm)somatic embryogenesis.I.Inittiation of embryogenic cultures from immature zygotic embryos[J].New For,1995;10:145-163.
[34]杨利平,程井辰。植物体细胞胚胎发生中的生理生化研究进展[J]。植物学通报,1991,8(2):1-8
[35]郭祖宝,王成增,孙培良。植物体细胞胚胎发生中的生理生化变化[J]。安徽农学通报,2007,13(14):52-53
[36]周燕,高述民等。胡萝卜体细胞胚胎发生中的细胞组织化学和蛋白质组分变 化[J]。植物生理学通讯,2004,40(2):181-183
[37]高述民,陆帼一,杜慧芳等。大蒜体细胞胚发育分化中特异蛋白质和某些生理生化变化[J]。植物生理学通讯,2001,37(3):207-23
[38]贺晓,慈忠玲等。紫花苜蓿体细胞胚胎发生及其细胞组织学观察[J]。内蒙古林学院学报(自然科学版),1999,21(2):44-46
[39]徐竹绮,王彤宁,何美瑛,等.水稻离体体细胞胚与合子胚的某此生化特性的比较[J].植物生理学报,1995,21(1):95-102.
[40]Kamada H.Harada H.Changes in endogenous amino acid compositions during somatic embryogenesis in Daucus carota L[J].Plant & Cell Physiol,1984,25:27-28
[41]杨和平,程井辰.马唐胚性和非胚性愈伤组织生理差异的初步研究[J]。植物生理学通讯,1991,27:337-34
[42]王亚馥,崔凯荣等。小麦体细胞胚发生的蛋白质组分和过氧化物酶同工酶的变化[J]。兰州大学学报(自然科学版),1993,29(3):189-193
[43]郑晓峰,黄百渠,几种植物体细胞胚胎发生标记蛋白的研究[J]。植物学报,1994,36(3):175-180
[44]Stirn S Jacobsen H.J.Marker proteins for embryogenic differentiation patterns in pea callus.Plant Cell,1987(6):50-54
[45]Chen L.J.Lathe D.S.Analysis of proteins from embryogenic and non-embryogenic rice(Oryza satire L·)calli·Plant Sci·1987,48:181-188
[46]崔凯荣,邢更生,周功克,等.体细胞胚发生的生化基础[J].生命科学,2001,13(1):28-33.
[47]戴若兰,张玮,邢更生,等.枸杞体细胞胚发生中DNA代谢动态的立体计量[J].植物学通报,2000,17(1):80-83
[48]Fujimura T,Komamine A,Matsumoto H.Aspects of DNA,RNA and protein synthesis during somatic embryogenesis in a carrot cell suspension culture. Physiol Plant,1980(49):255-260
[49]Ashibara H Z.Pflartzen et al.,1981,physil.,104:129-137
[50]Seugupta,Cand V.RagLavan,K.Exp.Bot.,1980(31):217-258
[51]Sevgupts,Cand V.Raghavan,K.Exp.Bot.,1980(31):259-268
[52]徐竹绮.2,4-D浓度对胡萝卜体细胞胚胎发生及其同工酶的影响[J].植物生理学报,1984,10(4):373-379
[53]Chibbar R.N et al.[J].Plant Physicl.,1988,133:367-370
[54]余沛涛,薛应龙.植物苯丙氨酸解氨酶(PAL)在细胞分化中的作用[J].植物生理学报,1987,13(1):14-19
[55]李国珍,秦明波,孙成华,等.青蒿体细胞胚胎发生及遗传转化的研究[J].植物学通报 1992,9(增刊)97-98
[56]崔凯荣,刑更生,王亚馥,等.枸杞组织培养中抗氧化酶活性与体细胞胚发生相关性的研究[J].兰州大学学报,1998,34(3):93-99.
[57]潘有福,王星,孟玉玲,等.秦艽体细胞胚不同发育阶段几种同工酶的特性[J].西北植物学报,1994,15(5):6-10
[58]陈耀锋,贺普超,廖祥和,等.葡萄脯氨酸累积变异系CAT和SOD活性研究[J].西北农业大学学报.1998,26(1):36-40.
[59]陈善春,张进仁,吴安仁,等.柳橙胚性愈伤组织耐盐性研究[J].西南农业大学学报,1987,9(4):463-468.
[60]林定波,颜秋生,沈德绪.柑橘抗寒细胞变异的获得及其抗性遗传稳定性的研究[J].植物学报,1999,41(2):136-141.
[61]胡含,王恒立.植物细胞工程与育种[M].北京:北京工业大学出版社,1990.
[62]郭梅.甘薯胚性恳浮细胞培养中的体细胞无性系变异分析[D].北京:中国农业大学,2005
[63]De capite L,Action of light and temperature on growth of plant tissue culture in vitro[J].Am.J.Bot.,1955,42:869-873.
[64]Seibert M,Wetherbee P J,Job D D,The effects of light intensity and spectral quality on growth and shoot initiation in tobacco callus[J].Plant Physiol.,1975,56:130-139.
[65]宋云,唐崇钦,匡廷云,等.不同光质下生长的满江红叶绿体和共生者满江红鱼腥藻的吸收光谱和荧光光谱的研究[J].植物生理学报,1983,9(1):69-76
[66]陶汉之,王新长.茶树光合作用与光质的关系[J].植物生理学通讯,1989,25(1):19-23
[67]范慈惠.光质对植物愈伤组织的诱导和植株形成的影响[J].植物生理学通讯,1981,17(1):25-33
[68]倪德祥,易莹,张丕方,等.光质对凹叶景天愈伤组织和不定芽形成的效应[J].上海农业学报,1988,4(1):43-48
[69]王维荣,王咏冬,欧阳光察,等.光质对黄瓜及番茄愈伤组织培养中分化和有关酶的影响[J].植物生理学报,1991,17(2):118-124.
[70]许桂芳,董诚明,周吉源,等.不同光质对华黄芪愈伤组织诱导、增殖及器官分化的效应[J].华中师范大学学报(自然科学版),1994,28(4):533-537.
[71]熊丽,周吉源,殷荣华.光质对石刁柏愈伤组织培养中生长和过氧化物酶的影响[J].武汉植物学研究,1995,13(3):253-257.
[72]阎秀峰,王洋,尚辛亥,等.光强和光质对野外栽培高山红景天生物量和红景天甙含量的影响[J].生态学报,2004,24(4):674-679
[73]赵德修,李茂寅,邢建民,等.光质、光强和光期对水母雪莲愈伤组织生长和黄酮生物合成的影响[J].植物生理学报,1999,25(2):127-13
[74]沈红香,沈漫,程继鸿,等.不同光质补光处理对郁金香生长和开花的影响[J].北京农学院学报,2007,22(1):16-18
[75]何小弟,裴建文,黄春华.菊花组织培养中的光质效应研究[J].江苏林业科技,2002,29(6):26-28
[76]周波,李玉花.植物的光敏色素与光信号转导[J].植物生理学通讯,2006, 42(1):134-140
[77]徐根娣,李瑶.光质和碳素对旱芹体细胞胚发生的影响[J].浙江师大学报,1995,18(2):57-60
[78]洪森荣,尹明华.光质对野葛叶片愈伤组织的诱导及其可溶性蛋白含量的影响[J].安徽农业科学,2007,35(9):2560-2563
[79]毛学文,陈荃.不同光质对毛地黄愈伤组织诱导和增值的效应[J].植物学通报,1997,14(1):55-56
[80]张君诚.银杏愈伤组织诱导与增殖的效应研究[J].重庆师范学院学报(自然科学版),1999,16(3):41-48
[81]梁钾贤,陈彪.光质对甘蔗愈伤组织组织分化出苗的影响[J].中国糖料,2006(3):9-11.
[82]Rey M,Diaz S,Rodriguez R.Exogenous polyamines imp rove rooting of hazelmicroshoots[J].Plant Cell Tissue Ore.Culture,1994,36(3):303-308
[83]倪德祥,陈刚,张丕方,等.锦葵愈伤组织培养中光质对其过氧化物同工酶和发根的效应[J].复旦学报(自然科学版),1986,25(2):157-162.
[84]倪德祥,王维荣,张丕方,等.菊花器Ⅱ光质和组合激素对菊花茎段培养中生长发育的影响[J].西北植物学报,1988,8(1):16-27
[85]李韶山,潘瑞炽.植物的蓝光效应[J].植物生理学通讯,1993,29(4):248-252
[86]王学奎,李合生,刘武定.光对小麦叶片谷氨酰胺合成酶调节机理初探[J].华中农业大学学报,2000,19(2):102-105
[87]王莉,史玲玲,刘玉军.不同光质对长鞭红景天悬浮细胞生长及苯丙氦酸解氨酶活性的影响[J].林业科学,2007,43(6):52-56
[88]Anna B,Alicjia K.Effect of light quality on somatic embryogenesis in Hyacinthus orientalis L.'Delft's blue'[J].Biological Bulletin of Poznan,2001,38(1):103-107
[89]梁学芬,蚁伟南,颜梓兴,等.不同光质的辅助光对香蕉组培苗的波长的影响[J].中国南方果树,2001,30(4):34
[90]Kasemir H.,Obdorfer U.,Mohr H.A twofold action of phytochrome in controlling chlorophyll a accumulation[J].Photochem,Photobiol,1973,18:481-486
[91]倪德祥,王蓓,王维荣,等.不同波长的光对佛甲草离体茎段培养过程中生长发育的影响[J].云南植物研究,1988,10(1):113-116
[92]Singla N K,Jain V,Jain S,et al.Activities of glycolytic enzymes in leaves and roots of contrasting cultivars of sorghum during flooding[J].Biologia Plantarum,2003,47(4):555-560
[93]Ferriol M,Pico B,Nuez F.Genetic diversity of a germplasm collection of cucurbita pepo using SRAP and AFLP markers[J].Heor Appl Genet,2003,107(2):271-282.
[94]倪德祥,冯文煦,张丕方,等.光质对一品红叶离体培养中形态发生及生化变化的影响[J].广西植物,1987,7(4):339-345
[95]倪德祥,易莹,张丕方,等.光质对凹叶景天愈伤组织和不定芽形成的效应[J].上海农业学报,1988,4(1):43-48
[96]Chen W,Cun SX.In vitro rapid propagation of stems of Dendrobium candidum.Plant Physiol.Commun.2002,(38):145.
[97]Shiau YJ,Nalawade SM,Hsia CN,Mulabacal V,Tsay HS.In Vitro propagation of the Chinese medicinal plant,Dendrobium Candidum Wall.Ex Lindl.,from axenic nodal segments.In Vitro Cell.Dev.Biol-Plant.2005,(41):666-670.
[98]He JB,Zheng CZ,Wang SL.Multiplication of protocorm of Dendrobium candidum.Acta.Botanica Yunnanica.1982,(4):211-212.
[99]Ye XL,Cheng SJ,Wang F,Qian NF.Morphology of immature seeds and development in vitro of Dendrobium candidum.Acta.Botanica Yunnanica.1988,(10):285-290.
[100]Zhang QX,Fang YM.Tissue culture and in vitro seedling and protocorm-like body examination of Dendrobium candidum.Acta.Botanica Boreali-Occidentalia Sinica.2005,(25):1761-1765.
[101]ZhaoP,Wang W,Feng FS,Wu F,Yang ZQ,Wang WJ.High-frequency shoot regeneration through transverse thin cell layer culture in Dendrobium Candidum Wall Ex Lind,Plant Cell,Tissue and Organ Culture,2007(90):131-139
[102]李阳春,谢可军.早熟禾属10种植物酯酶同工酶分析[J].草地学报,2003,11(3):214-219
[103]Hazard L,Ghesouiere M.Evidence from the use of isozyme markers of competition in swards between short leaved and long-leaved perennial ryegrass[J].Grass and Forage Science,1995,50:241-248
[104]Liu TL,Jiang XW,Wang LS.The analysis of special proteins and isoenzymes in globular embryogenesis of alfalfa in tissue culture.Acta.Botanica Boreali-Occidentalia Sinica.2002,(3):625-628.
[105]Everett NP,Wach MJ,Ashworth DJ.Biological marders of embryogenesis in tissue cultures of the mazie inbred in B73.Plant Sci.1985,(41):133-140
[106]Pan YF,Wang X,Meng YL,Wang LS.Characterization of several isoenzymes from embryogenic calli and somatic embryos at different developmental stages Gentiana Crassicaulis.Acta.Botanica Boreali-Occidentalia Sinica.1994,(15):6-10.