半夏试管小块茎的发育及其相关基因差异表达的研究
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摘要
半夏(Pinellia ternata Thunb.Briet)根状茎中含有半夏蛋白、鞣质和生物碱等药用成分,是我国一种特有的药用植物。半夏因病毒而引起的品种退化常导致种源不足、产量与品质严重下降,这已成为半夏生产上的主要障碍之一,通过茎尖分生组织脱除病毒培养的半夏苗和试管块茎等生物技术手段是生产上提供人工种子和提高产量品质的重要解决途径。半夏试管块茎是组织培养条件下诱导的变态茎,其遗传稳定性、生理生化特性上与常规块茎无异,半夏生产研究对此特别重视,但其试管小块茎的形成机理尚不清楚,生产效率不高,应用上尚存在着许多困难。本研究旨在添加不同内源激素抑制剂,研究半夏试管小块茎形成过程中内源激素的变化状况与小块茎形成相关基因的表达模式,探索块茎形成的调控机理,为半夏试管块茎生产提供理论和技术基础。主要研究结果如下:
通过MS基本培养基中不同浓度GA抑制剂(矮壮素,CCC)处理,对半夏试管块茎膨大和内源激素含量有一定影响。在半夏试管块茎形成过程中,CCC能有效的增加块茎重量,抑制GA_3,IAA,ZR的合成,并呈现出随着药剂浓度的增大而降低的趋势。CCC还能有效地影响ABA和JA含量,使之分别呈“V”型和“J”型。
通过MS基本培养基中不同浓度ABA抑制剂(哒草伏,NOR)处理,对半夏试管块茎膨大和内源激素含量有明显影响。在半夏试管块茎形成过程中,NOR能有效的减少块茎重量,抑制ABA,IAA,JA的合成,使JA含量变化呈“N”型。NOR还能有效地影响GA_3和ZR含量。
通过MS基本培养基中不同浓度JA抑制剂(水杨基异羟肟酸,SHAM)处理,对半夏试管块茎膨大和内源激素含量有明显影响。在半夏试管块茎形成过程中,SH AM能有效的推迟半夏块茎形成时间,促进在块茎形成后期GA_3,IAA,ZR的合成,但都低于对照。SHAM有效抑制JA的合成,同时还能有效地影响ABA,使之呈下降趋势。
提出了一种从半夏块茎中有效地提取高质量总RNA的方法。将常规的异硫氰酸胍提取植物总RNA的方法做了改进,建立起一种简单实用的从富含多酚类和多糖等物质的植物材料中提取总RNA的方法。该方法所提RNA的A_(260)/A_(230)均大于2.0,A_(260)/A_(280)的值为1.7～2.0,电泳条带清晰,RNA完整性好。利用该方法提取的半夏试管小块茎总RNA具有很高的质量和纯度,且得率很高,完全满足后续的DDRT-PCR分子生物学研究。
建立并优化了半夏块茎诱导前后银染mRNA差别显示条件。利用正交试验设计,从模板、Mg~(2+)、dNTPs、引物和DNA聚合酶5种因素4个水平,筛选并建立了半夏试管小块茎诱导的DDRT-PCR反应最佳体系。得到稳定且扩出最多条带的适合半夏试管小块茎诱导的DDRT-PCR最佳反应体系,即20μL的反应体系中含有dNTPs 150μmol/L,Taq酶0.6 U,锚定引物2μmol/L,随机引物1μmol/L,Mg~(2+)2.5mmol/L,2μL 10×buffer和2.5μg的cDNA模板。各因素影响程度依次为Taq酶浓度＞cDNA模板浓度＞dNTPs浓度＞Mg~(2+)浓度＞引物浓度。
利用mRNA差异显示技术分离和克隆了半夏试管小块茎发育的相关基因片段。应用mRNA差异显示(DDRT-PCR)方法比较分析了半夏试管小块茎发育过程中的基因表达差异。结果分离到了15个半夏试管小块茎发育过程中特异表达的cDNA片段,在GenBank中进行同源性比较发现后6个没有同源序列与之匹配;前3个片段推导的氨基酸序列分别与真核生物启动子,MADS-box蛋白和乙烯信号转导因子有高度同源性。半定量RT-PCR进一步证实它们存在,并在半夏生成不同天数高度特异表达。其中,这三个片段所推定的氨基酸都与块茎的发育有关,这为探讨半夏块茎发育分子机制提供了新资料。
Pinellia ternata(Thunb.) Briet.enriched with the medicinal ingredients of protein, tannim and alkaloid is an important steroid medicine resources plant that is endemic to China.The degeneration in quality of Pinellia ternata,caused by virus diseases usually results in a serious loss of both tuber yield and quality,which has been a main hinder in production of PinelIia ternata,for long time.Virus-elimination through meristem culture has been widely used in the seedling and microtubers of Pinellia ternata.and the development of biotechnology is important channels to resolve the production of artificial seeds and the improvment yield and quality.The microtuber of Pinellia ternata.,which is induced from the upper of leaf stalks ends,has the same genetic stability,morphological and physiological characteristics compared with conventional tubers.Perspectives of using microtubers of Pinellia ternata,in seed system has,therefore,drawn great interests in research.However,the productivity of microtubers is relatively low since there is little information about the tuberization mechanism.With aims to look into this aspect,the experiments were laid out under various tuber inducing conditions and investigations of variation in endogenous hormones and expression of tuber-forming-relative-genes (TFRG) were carried out.The main results were as follows:
Studied the effects of the inhibitor of gibberellins-biosynthesis on tuber enlargement and content of endogenous hormones of Pinellia ternata.Pinellia ternata microtubers were induced in the medium MS added with different concentration of CCC.The results showed that CCC could significantly add microtuber weight and inhibit the synthesis of GA_3,IAA,ZR and they were reduced as the content of CCC increased.Meanwhile,CCC could effectively influence the content of ABA and JA shaped as "V" and "J".
Studied the effects of the inhibitor of abscissas acid -biosynthesis on tuber enlargement and content of endogenous hormones of Pinellia ternata.Microtubers were induced in the medium MS added with different concentration of NOR.The results showed that NOR could significantly reduce microtuber weight and inhibit the synthesis of ABA,IAA,JA and the content of JA shaped as"N".Meanwhile,NOR could effectively influence the content of GA_3 and ZR.
Studied the effects of the inhibitor of jasmines acid-biosynthesis on tuber enlargement and content of endogenous hormones of Pinellia ternata.Microtubers of Pinellia ternata were induced in the medium MS added with different concentration of SHAM.The results showed that SHAM could significantly postponed the formation of microtuber time and promote the synthesis of GA_3,IAA,ZR,but the content of them were lower than collation.SHAM could effectively inhibited the synthesis of JA and decreased the content of ABA.
A highly efficient method of isolating total RNA from microtubers of Pinellia ternata in vitro was developed in this study.By modifying the method recommended by Guanidinium for extracting total RNA from plant tissues rich in phenolic and polysaccharidic compounds,a simple and convenient method for extraction of total RNA from plant materials containing abundant polyphenols and polysaccharides was established.The value of RNA extracted with improved method A260/ A230 were all over 2.0 and the values of A260/ A280 were between 1.7 and 2.0.The electrophoresis bands cleared on agarosegel and integrity of RNA was good.The results showed that RNA obtained from rapeseed with this method had high purity and quality and could be used in molecular biological research,as DDRT-PCR and analysis directly.
Establishment and optimization of sliver staining Differential Display of Pinellia ternate microtubers in vitro.The effects of five kinds of factors were studied by orthogonal design method including emplate cDNA,Mg~(2+),dNTPs,primers and Taq DNA polymerase,and in order to establish the optimum DDRT-PCR system of P.ternata. microtubers in vitro.A satisfactory DDRT-PCR technique system for P.ternata. Microtubers in vitro with desirable repeatability and polymorphic bands was established. In a total volume of 20μL DDRT-PCR system,it contained 10×buffer,150μmol/L dNTPs,2μmol/L anchor primer,1μmol/L arbitrary primer,2.5 mmol/L Mg~(2+),0.6 U Taq DNA polymerase and 2.5μg template cDNA.The effect of the five factors was in sequence of Taq DNA polymerase＞template cDNA＞dNTPs＞Mg~(2+)＞Primers.
Identification of expressed gene fragments related to the microtubers develop- ment of Pinellia ternat in vitro,by DDRT-PCR.In this study,we identified and characterized eight cDNA fragments differentially expressed in the induction of microtubers of Pinellia ternata in vitro,using mRNA differential display DDRT-PCR.Deduced amino -acid sequences of five of the eight cDNA fragments showed no significant homology with ESTs and genes in the Genbank databases.However,the remaining three showed significant homologies with sequences encoding components of eukaryotic translation initiation factor 3 subunit H,MADS-box protein and ethylene signal transcription factor, respectively.Their differential expression patterns were confirmed by semi-quantitative RT-PCR analysis.And the expression level of the induction of microtubers of Pinellia ternata in vitro.were different.The three cDNA fragments deduced like protein were most likely involved in the microtubers development of Pinellia ternata,and the study gave us novel insights into the molecular mechanism of the microtubers development of Pinellia ternata.
通过MS基本培养基中不同浓度GA抑制剂(矮壮素,CCC)处理,对半夏试管块茎膨大和内源激素含量有一定影响。在半夏试管块茎形成过程中,CCC能有效的增加块茎重量,抑制GA_3,IAA,ZR的合成,并呈现出随着药剂浓度的增大而降低的趋势。CCC还能有效地影响ABA和JA含量,使之分别呈“V”型和“J”型。
通过MS基本培养基中不同浓度ABA抑制剂(哒草伏,NOR)处理,对半夏试管块茎膨大和内源激素含量有明显影响。在半夏试管块茎形成过程中,NOR能有效的减少块茎重量,抑制ABA,IAA,JA的合成,使JA含量变化呈“N”型。NOR还能有效地影响GA_3和ZR含量。
通过MS基本培养基中不同浓度JA抑制剂(水杨基异羟肟酸,SHAM)处理,对半夏试管块茎膨大和内源激素含量有明显影响。在半夏试管块茎形成过程中,SH AM能有效的推迟半夏块茎形成时间,促进在块茎形成后期GA_3,IAA,ZR的合成,但都低于对照。SHAM有效抑制JA的合成,同时还能有效地影响ABA,使之呈下降趋势。
提出了一种从半夏块茎中有效地提取高质量总RNA的方法。将常规的异硫氰酸胍提取植物总RNA的方法做了改进,建立起一种简单实用的从富含多酚类和多糖等物质的植物材料中提取总RNA的方法。该方法所提RNA的A_(260)/A_(230)均大于2.0,A_(260)/A_(280)的值为1.7～2.0,电泳条带清晰,RNA完整性好。利用该方法提取的半夏试管小块茎总RNA具有很高的质量和纯度,且得率很高,完全满足后续的DDRT-PCR分子生物学研究。
建立并优化了半夏块茎诱导前后银染mRNA差别显示条件。利用正交试验设计,从模板、Mg~(2+)、dNTPs、引物和DNA聚合酶5种因素4个水平,筛选并建立了半夏试管小块茎诱导的DDRT-PCR反应最佳体系。得到稳定且扩出最多条带的适合半夏试管小块茎诱导的DDRT-PCR最佳反应体系,即20μL的反应体系中含有dNTPs 150μmol/L,Taq酶0.6 U,锚定引物2μmol/L,随机引物1μmol/L,Mg~(2+)2.5mmol/L,2μL 10×buffer和2.5μg的cDNA模板。各因素影响程度依次为Taq酶浓度＞cDNA模板浓度＞dNTPs浓度＞Mg~(2+)浓度＞引物浓度。
利用mRNA差异显示技术分离和克隆了半夏试管小块茎发育的相关基因片段。应用mRNA差异显示(DDRT-PCR)方法比较分析了半夏试管小块茎发育过程中的基因表达差异。结果分离到了15个半夏试管小块茎发育过程中特异表达的cDNA片段,在GenBank中进行同源性比较发现后6个没有同源序列与之匹配;前3个片段推导的氨基酸序列分别与真核生物启动子,MADS-box蛋白和乙烯信号转导因子有高度同源性。半定量RT-PCR进一步证实它们存在,并在半夏生成不同天数高度特异表达。其中,这三个片段所推定的氨基酸都与块茎的发育有关,这为探讨半夏块茎发育分子机制提供了新资料。
Pinellia ternata(Thunb.) Briet.enriched with the medicinal ingredients of protein, tannim and alkaloid is an important steroid medicine resources plant that is endemic to China.The degeneration in quality of Pinellia ternata,caused by virus diseases usually results in a serious loss of both tuber yield and quality,which has been a main hinder in production of PinelIia ternata,for long time.Virus-elimination through meristem culture has been widely used in the seedling and microtubers of Pinellia ternata.and the development of biotechnology is important channels to resolve the production of artificial seeds and the improvment yield and quality.The microtuber of Pinellia ternata.,which is induced from the upper of leaf stalks ends,has the same genetic stability,morphological and physiological characteristics compared with conventional tubers.Perspectives of using microtubers of Pinellia ternata,in seed system has,therefore,drawn great interests in research.However,the productivity of microtubers is relatively low since there is little information about the tuberization mechanism.With aims to look into this aspect,the experiments were laid out under various tuber inducing conditions and investigations of variation in endogenous hormones and expression of tuber-forming-relative-genes (TFRG) were carried out.The main results were as follows:
Studied the effects of the inhibitor of gibberellins-biosynthesis on tuber enlargement and content of endogenous hormones of Pinellia ternata.Pinellia ternata microtubers were induced in the medium MS added with different concentration of CCC.The results showed that CCC could significantly add microtuber weight and inhibit the synthesis of GA_3,IAA,ZR and they were reduced as the content of CCC increased.Meanwhile,CCC could effectively influence the content of ABA and JA shaped as "V" and "J".
Studied the effects of the inhibitor of abscissas acid -biosynthesis on tuber enlargement and content of endogenous hormones of Pinellia ternata.Microtubers were induced in the medium MS added with different concentration of NOR.The results showed that NOR could significantly reduce microtuber weight and inhibit the synthesis of ABA,IAA,JA and the content of JA shaped as"N".Meanwhile,NOR could effectively influence the content of GA_3 and ZR.
Studied the effects of the inhibitor of jasmines acid-biosynthesis on tuber enlargement and content of endogenous hormones of Pinellia ternata.Microtubers of Pinellia ternata were induced in the medium MS added with different concentration of SHAM.The results showed that SHAM could significantly postponed the formation of microtuber time and promote the synthesis of GA_3,IAA,ZR,but the content of them were lower than collation.SHAM could effectively inhibited the synthesis of JA and decreased the content of ABA.
A highly efficient method of isolating total RNA from microtubers of Pinellia ternata in vitro was developed in this study.By modifying the method recommended by Guanidinium for extracting total RNA from plant tissues rich in phenolic and polysaccharidic compounds,a simple and convenient method for extraction of total RNA from plant materials containing abundant polyphenols and polysaccharides was established.The value of RNA extracted with improved method A260/ A230 were all over 2.0 and the values of A260/ A280 were between 1.7 and 2.0.The electrophoresis bands cleared on agarosegel and integrity of RNA was good.The results showed that RNA obtained from rapeseed with this method had high purity and quality and could be used in molecular biological research,as DDRT-PCR and analysis directly.
Establishment and optimization of sliver staining Differential Display of Pinellia ternate microtubers in vitro.The effects of five kinds of factors were studied by orthogonal design method including emplate cDNA,Mg~(2+),dNTPs,primers and Taq DNA polymerase,and in order to establish the optimum DDRT-PCR system of P.ternata. microtubers in vitro.A satisfactory DDRT-PCR technique system for P.ternata. Microtubers in vitro with desirable repeatability and polymorphic bands was established. In a total volume of 20μL DDRT-PCR system,it contained 10×buffer,150μmol/L dNTPs,2μmol/L anchor primer,1μmol/L arbitrary primer,2.5 mmol/L Mg~(2+),0.6 U Taq DNA polymerase and 2.5μg template cDNA.The effect of the five factors was in sequence of Taq DNA polymerase＞template cDNA＞dNTPs＞Mg~(2+)＞Primers.
Identification of expressed gene fragments related to the microtubers develop- ment of Pinellia ternat in vitro,by DDRT-PCR.In this study,we identified and characterized eight cDNA fragments differentially expressed in the induction of microtubers of Pinellia ternata in vitro,using mRNA differential display DDRT-PCR.Deduced amino -acid sequences of five of the eight cDNA fragments showed no significant homology with ESTs and genes in the Genbank databases.However,the remaining three showed significant homologies with sequences encoding components of eukaryotic translation initiation factor 3 subunit H,MADS-box protein and ethylene signal transcription factor, respectively.Their differential expression patterns were confirmed by semi-quantitative RT-PCR analysis.And the expression level of the induction of microtubers of Pinellia ternata in vitro.were different.The three cDNA fragments deduced like protein were most likely involved in the microtubers development of Pinellia ternata,and the study gave us novel insights into the molecular mechanism of the microtubers development of Pinellia ternata.
引文
1.安丰英,郭红卫.乙烯信号转导的分子机制.植物学通报,2006,23(5):531-542
2.蔡小钿,王金发.植物MADS盒基因的功能和调节机理.植物生理学通讯,2000,36:277-281
3.曹锫生,蔡汗.羊葬球茎离体诱导技术的研究.园艺学报,1999,26(5):335-336
4.常莉.半夏珠芽的位置效应及其影响因素[硕士学位论文].武汉:华中农业大学图书馆,2007
5.常莉,徐有明,薛建平.离体培养条件下半夏叶柄形成珠芽过程中内源激素的变化.华中农业大学学报,2007,26(5):612-615
5.陈涛,张劲松.乙烯的生物合成与信号传递.植物学通报,2006,23(5):519-530
6.崔瑾,李式军.水杨酸对芋试管球茎发育的影响.南京农业大学学报,2003,26(1):97-99
7.崔凯荣,邢更生,秦琳,王亚馥.利用mRNA差别显示技术分析枸杞体细胞胚发生早期基因的差别表达.遗传,1998,20(5):16-19
8.丁勇,刘英,杨鸯鸯,甘莉.油菜种子高质量总RNA提取的一种有效方法.华中农业大学学报,2006,25(5):465-468
9.杜长玉,李东明,张志龙.不同生长素在马铃薯上应用效果的研究.中国马铃薯,2000,14(3):137-140.
10.方孝东,林栖风,李冠一.对代表性差异分析方法的改进.海南大学学报(自然科学版),2003,21(2):135-138.
11.甘立军,曾晓春,周燮.茉莉酸类与植物地下贮藏器官的形成.植物学通报,2001,18(5):546-553
12.郭宾会.小麦苗期水分胁迫诱导差异表达cDNA的研究.生物技术通报,2003,2:40-44
13.郭永兵,张友德.盾叶薯裁根茎中薯孩皂昔元的分布.湖北农业科学,2002,(5):106-107
14.韩德俊,陈耀锋,李春莲.水杨酸对马铃薯试管微薯形成的影响研究.西北植物学报,1999,19(3):428-433.
15.韩献忠,张治国.半夏组织培养一次性成苗的研究.中草药,1989,20(11):45
16.郝纯毅.降低mRNA差异显示技术假阳性率的一种方法.中国生物化学与分子生学报,2002,18(1):110-114
17.何奕昆,刘刚,路铁刚,孙敬三.半夏茎尖培养及块茎的品质改良.植物学报,1994,3(1):39-44
18.何祖华,吴玉良,董继新,董海涛,李德葆.分子生物学新方法.生命科学,1999,11(增刊):129-131
19.胡丽芳,金志强,徐碧玉.MADS-box基因在果实发育、成熟过程中的作用.分子植物育种,2005,3(3):415-420
20.黄旭.mRNA差异显示法分离水稻抗稻瘟病相关cDNA克隆.江苏农业学报,2000,16(2):124-126
21.江树业,陈启锋,方宣钧.水稻光(温)敏核不育基因的定位分离、研究进展及对策.高技术通讯,1998,2:55-58
22.江树业,吕蓓,陈启锋,方宣钧.籼型光敏核不育水稻Hs-1可育和不育幼穗mRNA的差异达分析.农业生物技术学报,2001,9(1):69-71
23.李春香,杨军,王树才,夏凯,李式军,周燮.水杨酸在大蒜鳞茎膨大中的作用.园艺学报,2000,27(3):220-222
24.李春香,周燮.MeJA对大蒜鳞茎膨大及内源激素含量的影响.生命科学研究,2002,6(2):183-183.
25.李德红,潘瑞炽.水杨酸在植物体内的作用.植物生理学通讯,1995,31(2):144-149
26.李亮亮,朱宝成,成亚利,吴爱民,李庆余.掌叶半夏组织培养过程中的染色体变异.农业生物技术学报,1998,6(1):23-27
27.李玉京,李子银,李振声.真核生物mRNA差显技术(Differ-ential Display)的研究进展.生物技术通报,1998,(5):23-30
28.李宗霆,周燮.植物激素及其免疫检测技术.南京:江苏科技出版社,1996:225-226
29.连勇,邹颖,东惠茹,金黎平,林桓.马铃薯试管薯形成过程中几种内源激素的变化.园艺学报,2002,29(6):537-541
30.刘梦芸,蒙美莲,门福义,毛雪飞.GA_3、IAA、CTK和ABA对马铃薯块茎形成调控作用的研究.内蒙古农牧学院学报,1997,18(2):16-20
31.柳俊,谢从华.马铃薯试管块茎发育机制及相关基因表达.植物学通报,2001,18(5):531-539
32.柳俊.马铃薯试管块茎的形成机理及块茎形成调控[博士学位论文].武汉:华中农大图书馆,2001
33.罗光明.半夏的组织培养和植株再生.中药研究与信息,2000,2(4):13-14
34.马崇坚.马铃薯试管块茎形成与内源生长物质的关系[博士学位论文].武汉:华中农业大学图书馆,2003
35.毛子成,彭正松.半夏快速繁殖体系的研究进展.中国中药杂志,2003,28(3):193-195
36.蒙美莲,刘梦芸,门福义,陈有君.马铃薯块茎生长过程中内源赤霉素和脱落酸含量的变化.马铃薯杂志,1996,10(1):8-12
37.蒙美莲,刘梦芸,门福义,张宏志.赤霉素和脱落酸对马铃薯块茎形成的影响.铃薯杂志,1994,8(3):134-137
38.倪中福,孙其信,吴利民.普通小麦不同优势杂交种及其亲本之间基因表达差异比较研究.中国农业大学学报,2000,5(1):1-8
39.齐小红.用DDRT-PCR技术筛选仔猪腹泻候选基因.中国农业大学博士学位论文,2003
40.全峰,张爱霞,曹先维.植物激素在马铃薯块茎发育过程中的作用.中国马铃薯,2002,16 (1):29-31
41.沈淞海,郭奇珍.茉莉酮酸类物质与块茎形成.植物生理学通讯,1996,32(4):292-294
42.孙红梅,李天来,李云飞.低温解除休眠过程中兰州百合中部鳞片物质变化的生理机制.中国农业科学,2005,38(2):376-382
43.田振东,柳俊,谢从华,宋波涛.DDRT-PCR与反向Northern杂交技术结合分离马铃薯mRNA 差异表达片段.农业生物技术学报,2003,11(5):545-546
44.王芳,张斌,单雷,毕玉平,许泽永.一种从花生种子中提取RNA的改进方法.花生学报,2002,31(4):24-26
45.王光清,胡建广,赵相山,程宁辉,钱晓茵,杨金水.水稻愈伤组织形态发生中的MADS盒基因的差异表达.植物学报,1997,39(1):1035-1041
46.王庆美,张立明,王振林.甘薯内源激素变化与块根形成膨大的关系.中国农业科学,2005,38(12):414-420
47.王三根.细胞分裂素在植物抗逆和延衰中的作用.植物学通报,2000,17(2):121-126
48.夏英,杨梅英.用PCR-SSCP法鉴定差异显示中的正确的阳性克隆cDNA片段.中国辐射卫生,2002,11(1):40
49.肖华胜,黄文晋.降低差异显示PCR假阳性和提高重复性的几种策略.生物化学展,1999,26(5):437-438
50.熊正琴,李式军,周燮.茉莉酸甲酷和水杨酸促进大蒜试管鳞茎的形成.园艺学报,1999,26(6):408-409
51.薛建平,张爱民,盛玮,翟志军.钾盐对半夏试管块茎诱导的影响.中国中药杂志,2006,31(7):546-548
52.薛建平,朱艳芳,张爱民,柳俊.半夏试管块茎直接再生技术的研究.作物学报,2004,30(10):1060-1064
53.杨亮,付丽娅,刘仲齐,林伟利,张秀明,乔明强,白艳玲.富含多糖番茄果实组织中总RNA的有效提取方法.南开大学学报(自然科学版),2005,38(5):36-39
54.曾令波,周吉源,赵洁,熊丽.半夏组织培养中不同植物生长调节物质对形态发生的效应.安徽师范大学学报(自然科学版),1995,18(3):42-46
55.曾英,胡金勇,李志坚.植物MADS盒基因与花器官的进化发育.植物生理学通讯,2001,37:281-287
56.张驰,陈受宜.利用DDRT-PCR技术分析在盐胁迫下水稻耐盐突变体中特异表达的基因.中国科学(B辑),1995,25(8):840-847
57.张洪涛,张春燕,杨秋格,杨学惠.银染mRNA差异显示方法的建立和二次扩增参数优化.癌症,2001,20(3):330-331
58.张景凤,赵慧,桂晋刚,刘坤凡,王道文.利用RT-PCR、DNA序列分析及原核基因表达对我国大豆花叶病毒进行分子鉴定.植物学报,1999,41(9):932-935
59.张丽梅,陈替瑛,陈瑞云.半夏组织培养一次性成苗技术研究初报.福建农业科技,2003,1:20
60.张苏锋,谢素霞.半夏组织培养快速繁殖的研究.信阳师范学院报,1998,11(1):86-88
61.张则婷,李学宝.MADS-box基因在植物发育中的功能.植物生理学通讯,2007,43(2):218-222
62.赵大中.运用差异显示法分离冬小麦春化作用相关cDNA克隆.科学通报,1998,43(9):965-968.
63.赵月玲,夏海武,万永霞.活性炭在半夏组织培养中的使用.昌潍师专学报,1999,18(2):15-17
64.朱忠荣,下永树,杨业正,雷耀森,雷元林.三叶半夏组织培养快速繁殖研究.山地农业生物学报,1991,10(1):40-45
65.J萨姆布鲁克,D W拉塞尔著,黄培堂译.分子克隆实验指南.北京:科学出版社,2003,1688
66.App leyard V C L,U nk les S E.Secondary metabo lite production in filamenfous fungi disp layed.Mo 1 GenGenet,1995,247:338-342.
67.Arnold A M,Anderson G W,McIver B et al.A novel dynamin Ⅱ Iisoform is up-regulated in the cent ral nervous system in hypothyroidism.Int J Deve Neuroscience,2003,21:267-275
68.Bachem C W B,Hoevena V A N,Ded R S.Visualization of differential gene expression using a novel method of RNA fingerprinting based on AFLP:analysis of gene expression during potato tuber developing.Plant Jour,1996,9(5):745-754
69.BAUER D,MULLER H,REICH J.Identification of Differentially Expressed mRNA Species by an Imp roved Display Technique(DDRT-PCR).Nucleic Acid Res,1993,21(18):4 272-4280.
70.Benito E P,Prins T,Kan J A L.Application of differential display RT-PCR to the ananly-sis of gene expression in a plant-fungus interaction.Plant Mol Biol,1996,32:947-957.
71.Bleecker,A.B.and Kende,H..Ethylene:agaseous signal molecule in plants.Annu.Rev.Cell Dev.Biol.2000,16,1-18.
72.Carrera E,Bou J,Garcia-Martinez J L.Changes in GA 20-oxidase gene expression strongly affect stem length,tuber induction and tuber yield of potato plant.Plant J,2000,22:247-256.
73.Chen X F,W ang B Y,W u R.The level of a mRNA with sequence similarity to the old yellow enzyme-NADPH dehydro2genases increases in chenopodium rubrum cells in response to cytok inin.Jour Plant Physiol,1996,149(12):233-236.
74.DIACHENKO L B,LEDESMA J,CHENCHIKA A.Combing the Techniques of RNA Fingerp rinting and Differential Disp lay to Obtain Differentially Expressed mRNA.Biochem Biophys Res Commun,1996,219:824-828
75.Doss R P.Differential display without radioactivity-a modified procedure.BioTechniques,1996, 21 :410-412
76. Esther V D K, Kende H. Identification of a gibberellin induced gene in deepwater rice using differential display of mRNA. Plant Mol, 1995, 28: 589-592
77. Fang G, Hammar S, Grumet R. A quick and in expensive method for removing polysaccharides from plant genomic DNA. Bio Techniques, 1992, 13:52-55
78. FischerA, Saedler H, Theissen G. Restriction FragmentLength Polymorphism Coupled Domain Directed DifferentialDisplay: A Highly Efficient Technique for Expression Analysisof Multigene Families.Proceedings of the National Academy of Sciences of the United States of America, 1995, 92:5331-5335
79. Fujino K, Koda Y, Kikuta Y. Reorientation of cortical microtubules in the subapical region during tuberization in single-node stem segments of potato in culture. Plant Cell Physiol. 1995, 36: 891-895
80. Gang An, GuiZhen Luo, RobertW. Sensitive, Nonradioactive Differential Display Method Using Chemiluminescent Detection.Biology Techniques, 1996,20: 342 - 346
81. Hedden P. Regulation of gibberellin biosynthesis. Hooykaas P J J, Hall M A, Libbenga K R eds. Biochemistry and molecular biology of plant hormones. Elsevier Science: Amsterdam, 1999, 161-188
82. HUBANKM, SCHATZD G Identifying Difference in mRNA Expression by Reresentational Difference Analysis of cDNA. Nucl acids Res, 1994, 22 (25 ): 5 640 -5 648.
83. Jackson S D. Multiple signaling pathways control tuber induction in potato. Plant Physiol, 1999, 119:1-8
84. Jacobs A. Mo lecular analysis of cold resistance in ch ilean po tato species. Aminor field study. Work ing paper international rural development centre, Swedish U niversity of A griculture. Science, 1996, 305: 15-19
85. Kammer H V D , C Albrecht ,C Albrecht. Identification of genes regulated by muscarinic acetylcholine receptors: pplication of an improved and statistically comprehensive mRNA differential display technique. Nucleic Acids Research ,1999 , 27 (10) :2211-2218
86. Knaap E van der, Knende H. Identification of a gibberellin-induced gene in deepwater rice using differential-display of mRNA. PlantMo .Bio ., 1995, 28: 589- 592.
87. Koda Y. Possible involvement of jasmonates in various morphogenic events. Phsiol Plant, 1999, 100:639-646
88. Kolomiets M V, Hannapel D J, Chen H, Tymeson M, Gladon R J. Lipoxygenase Is Involved in the Control of Potato Tuber Development. The Plant Cell, 2001,13: 613-626
89. Lee S W, Tomasetto C , Sager R. Positive selection of candidate tumorsuppressor Genes by subtractive hybridization. Proc Natl Acad Sci USA, 1991 , 88 :2825 -2829
90. Leonard S. Jefferson Saot R. Kimball. Amino Acid Regulation of Gene Expression. Journal of Nutrition.2001,31:2460-2466
91. Liang P, Pardee A B. Differential Display of eukaryotic messenger RNA by means of the polymerase chain reaction. Science, 1992 , 257 (14): 961-971
92. LISITSYN N, W IGKER M. Cloning the Differences Betweentwo Complex Genomes.Science, 1993, 259 (5097) :946 - 951
93. Liu J, Xie C H, Liao Y, Huang D E. Effects of endogenous gibberellic acid and abscisic acid on the induction of potato microtubers in vitro. In: Liu Q and Kokubu J ads.Proceedings of 1st Chinese-Japanese Symposium on Sweetpotato and Potato. Beijing agricultural University, Beijing, 1995,267-275
94. Lohmann J, Schickle H, Bo sch TCG. REN display, a rapid and efficient method for nonradioactive differential display and mRNA isolation. Bio Tech, 1995; 18:200-202
95. Matz M and Lukyanov S A .Different strategies of differential display: areas of application. Nucleic Acids Res. 1998, 26(24):5537-5543
96. Mikhail Matz, Natalia Usman, Dmitryi Shagin.Ordered differential display: a simple method for systematic comparison of gene exp ression profiles.Nucleic Acids Research, 1997, 25: 2541 -2542.
97. Opsahl F H C , Dewnff E L, Dumas C. A novel endosperm-specific gene express -ed in a restricted region around the maize embryo. Plant Jour, 1997,12(1): 235 -246
98. Peng Liang, ArthurB Pardee.Differential Display of Eukaryotic Messenger RNA by Means of the Ploymerase Chain Reaction.Science, 1992, 257: 967 - 971
99. Radmacher W. Growth retardants: effects on gibberellin biosynthesis and other metabolic pathways. Annusl Review of Plant Physiology and Plant Molecular and biology,2000, 51:501-503
100. Robert P Doss.Differential Display Without Radioactivity A Modified Procedur.Biology Techniques, 1996, 21:408-412
101. Rossi Jaume A, Corral M, Tiai) R. Possible involvement of jasmonic acid in bulb formation of garlic (Allium satiuanl L). Acta Hort, 1997, 433:381-388
102. Sharma Y K, Davis K R. Iso lation of a novel A rabidop sis ozone2induced cDNA by differential display. Plant Mo.Bio., 1995, 29: 91- 98.
103. Simon HG, Oppenheimer S. A dvanced mRNA differential disp lay:isolation of a new differential regulated myosin heavy chain-ncoding gene in amphibian limb regeneration. Gene, 1996, 72: 175-181
2.蔡小钿,王金发.植物MADS盒基因的功能和调节机理.植物生理学通讯,2000,36:277-281
3.曹锫生,蔡汗.羊葬球茎离体诱导技术的研究.园艺学报,1999,26(5):335-336
4.常莉.半夏珠芽的位置效应及其影响因素[硕士学位论文].武汉:华中农业大学图书馆,2007
5.常莉,徐有明,薛建平.离体培养条件下半夏叶柄形成珠芽过程中内源激素的变化.华中农业大学学报,2007,26(5):612-615
5.陈涛,张劲松.乙烯的生物合成与信号传递.植物学通报,2006,23(5):519-530
6.崔瑾,李式军.水杨酸对芋试管球茎发育的影响.南京农业大学学报,2003,26(1):97-99
7.崔凯荣,邢更生,秦琳,王亚馥.利用mRNA差别显示技术分析枸杞体细胞胚发生早期基因的差别表达.遗传,1998,20(5):16-19
8.丁勇,刘英,杨鸯鸯,甘莉.油菜种子高质量总RNA提取的一种有效方法.华中农业大学学报,2006,25(5):465-468
9.杜长玉,李东明,张志龙.不同生长素在马铃薯上应用效果的研究.中国马铃薯,2000,14(3):137-140.
10.方孝东,林栖风,李冠一.对代表性差异分析方法的改进.海南大学学报(自然科学版),2003,21(2):135-138.
11.甘立军,曾晓春,周燮.茉莉酸类与植物地下贮藏器官的形成.植物学通报,2001,18(5):546-553
12.郭宾会.小麦苗期水分胁迫诱导差异表达cDNA的研究.生物技术通报,2003,2:40-44
13.郭永兵,张友德.盾叶薯裁根茎中薯孩皂昔元的分布.湖北农业科学,2002,(5):106-107
14.韩德俊,陈耀锋,李春莲.水杨酸对马铃薯试管微薯形成的影响研究.西北植物学报,1999,19(3):428-433.
15.韩献忠,张治国.半夏组织培养一次性成苗的研究.中草药,1989,20(11):45
16.郝纯毅.降低mRNA差异显示技术假阳性率的一种方法.中国生物化学与分子生学报,2002,18(1):110-114
17.何奕昆,刘刚,路铁刚,孙敬三.半夏茎尖培养及块茎的品质改良.植物学报,1994,3(1):39-44
18.何祖华,吴玉良,董继新,董海涛,李德葆.分子生物学新方法.生命科学,1999,11(增刊):129-131
19.胡丽芳,金志强,徐碧玉.MADS-box基因在果实发育、成熟过程中的作用.分子植物育种,2005,3(3):415-420
20.黄旭.mRNA差异显示法分离水稻抗稻瘟病相关cDNA克隆.江苏农业学报,2000,16(2):124-126
21.江树业,陈启锋,方宣钧.水稻光(温)敏核不育基因的定位分离、研究进展及对策.高技术通讯,1998,2:55-58
22.江树业,吕蓓,陈启锋,方宣钧.籼型光敏核不育水稻Hs-1可育和不育幼穗mRNA的差异达分析.农业生物技术学报,2001,9(1):69-71
23.李春香,杨军,王树才,夏凯,李式军,周燮.水杨酸在大蒜鳞茎膨大中的作用.园艺学报,2000,27(3):220-222
24.李春香,周燮.MeJA对大蒜鳞茎膨大及内源激素含量的影响.生命科学研究,2002,6(2):183-183.
25.李德红,潘瑞炽.水杨酸在植物体内的作用.植物生理学通讯,1995,31(2):144-149
26.李亮亮,朱宝成,成亚利,吴爱民,李庆余.掌叶半夏组织培养过程中的染色体变异.农业生物技术学报,1998,6(1):23-27
27.李玉京,李子银,李振声.真核生物mRNA差显技术(Differ-ential Display)的研究进展.生物技术通报,1998,(5):23-30
28.李宗霆,周燮.植物激素及其免疫检测技术.南京:江苏科技出版社,1996:225-226
29.连勇,邹颖,东惠茹,金黎平,林桓.马铃薯试管薯形成过程中几种内源激素的变化.园艺学报,2002,29(6):537-541
30.刘梦芸,蒙美莲,门福义,毛雪飞.GA_3、IAA、CTK和ABA对马铃薯块茎形成调控作用的研究.内蒙古农牧学院学报,1997,18(2):16-20
31.柳俊,谢从华.马铃薯试管块茎发育机制及相关基因表达.植物学通报,2001,18(5):531-539
32.柳俊.马铃薯试管块茎的形成机理及块茎形成调控[博士学位论文].武汉:华中农大图书馆,2001
33.罗光明.半夏的组织培养和植株再生.中药研究与信息,2000,2(4):13-14
34.马崇坚.马铃薯试管块茎形成与内源生长物质的关系[博士学位论文].武汉:华中农业大学图书馆,2003
35.毛子成,彭正松.半夏快速繁殖体系的研究进展.中国中药杂志,2003,28(3):193-195
36.蒙美莲,刘梦芸,门福义,陈有君.马铃薯块茎生长过程中内源赤霉素和脱落酸含量的变化.马铃薯杂志,1996,10(1):8-12
37.蒙美莲,刘梦芸,门福义,张宏志.赤霉素和脱落酸对马铃薯块茎形成的影响.铃薯杂志,1994,8(3):134-137
38.倪中福,孙其信,吴利民.普通小麦不同优势杂交种及其亲本之间基因表达差异比较研究.中国农业大学学报,2000,5(1):1-8
39.齐小红.用DDRT-PCR技术筛选仔猪腹泻候选基因.中国农业大学博士学位论文,2003
40.全峰,张爱霞,曹先维.植物激素在马铃薯块茎发育过程中的作用.中国马铃薯,2002,16 (1):29-31
41.沈淞海,郭奇珍.茉莉酮酸类物质与块茎形成.植物生理学通讯,1996,32(4):292-294
42.孙红梅,李天来,李云飞.低温解除休眠过程中兰州百合中部鳞片物质变化的生理机制.中国农业科学,2005,38(2):376-382
43.田振东,柳俊,谢从华,宋波涛.DDRT-PCR与反向Northern杂交技术结合分离马铃薯mRNA 差异表达片段.农业生物技术学报,2003,11(5):545-546
44.王芳,张斌,单雷,毕玉平,许泽永.一种从花生种子中提取RNA的改进方法.花生学报,2002,31(4):24-26
45.王光清,胡建广,赵相山,程宁辉,钱晓茵,杨金水.水稻愈伤组织形态发生中的MADS盒基因的差异表达.植物学报,1997,39(1):1035-1041
46.王庆美,张立明,王振林.甘薯内源激素变化与块根形成膨大的关系.中国农业科学,2005,38(12):414-420
47.王三根.细胞分裂素在植物抗逆和延衰中的作用.植物学通报,2000,17(2):121-126
48.夏英,杨梅英.用PCR-SSCP法鉴定差异显示中的正确的阳性克隆cDNA片段.中国辐射卫生,2002,11(1):40
49.肖华胜,黄文晋.降低差异显示PCR假阳性和提高重复性的几种策略.生物化学展,1999,26(5):437-438
50.熊正琴,李式军,周燮.茉莉酸甲酷和水杨酸促进大蒜试管鳞茎的形成.园艺学报,1999,26(6):408-409
51.薛建平,张爱民,盛玮,翟志军.钾盐对半夏试管块茎诱导的影响.中国中药杂志,2006,31(7):546-548
52.薛建平,朱艳芳,张爱民,柳俊.半夏试管块茎直接再生技术的研究.作物学报,2004,30(10):1060-1064
53.杨亮,付丽娅,刘仲齐,林伟利,张秀明,乔明强,白艳玲.富含多糖番茄果实组织中总RNA的有效提取方法.南开大学学报(自然科学版),2005,38(5):36-39
54.曾令波,周吉源,赵洁,熊丽.半夏组织培养中不同植物生长调节物质对形态发生的效应.安徽师范大学学报(自然科学版),1995,18(3):42-46
55.曾英,胡金勇,李志坚.植物MADS盒基因与花器官的进化发育.植物生理学通讯,2001,37:281-287
56.张驰,陈受宜.利用DDRT-PCR技术分析在盐胁迫下水稻耐盐突变体中特异表达的基因.中国科学(B辑),1995,25(8):840-847
57.张洪涛,张春燕,杨秋格,杨学惠.银染mRNA差异显示方法的建立和二次扩增参数优化.癌症,2001,20(3):330-331
58.张景凤,赵慧,桂晋刚,刘坤凡,王道文.利用RT-PCR、DNA序列分析及原核基因表达对我国大豆花叶病毒进行分子鉴定.植物学报,1999,41(9):932-935
59.张丽梅,陈替瑛,陈瑞云.半夏组织培养一次性成苗技术研究初报.福建农业科技,2003,1:20
60.张苏锋,谢素霞.半夏组织培养快速繁殖的研究.信阳师范学院报,1998,11(1):86-88
61.张则婷,李学宝.MADS-box基因在植物发育中的功能.植物生理学通讯,2007,43(2):218-222
62.赵大中.运用差异显示法分离冬小麦春化作用相关cDNA克隆.科学通报,1998,43(9):965-968.
63.赵月玲,夏海武,万永霞.活性炭在半夏组织培养中的使用.昌潍师专学报,1999,18(2):15-17
64.朱忠荣,下永树,杨业正,雷耀森,雷元林.三叶半夏组织培养快速繁殖研究.山地农业生物学报,1991,10(1):40-45
65.J萨姆布鲁克,D W拉塞尔著,黄培堂译.分子克隆实验指南.北京:科学出版社,2003,1688
66.App leyard V C L,U nk les S E.Secondary metabo lite production in filamenfous fungi disp layed.Mo 1 GenGenet,1995,247:338-342.
67.Arnold A M,Anderson G W,McIver B et al.A novel dynamin Ⅱ Iisoform is up-regulated in the cent ral nervous system in hypothyroidism.Int J Deve Neuroscience,2003,21:267-275
68.Bachem C W B,Hoevena V A N,Ded R S.Visualization of differential gene expression using a novel method of RNA fingerprinting based on AFLP:analysis of gene expression during potato tuber developing.Plant Jour,1996,9(5):745-754
69.BAUER D,MULLER H,REICH J.Identification of Differentially Expressed mRNA Species by an Imp roved Display Technique(DDRT-PCR).Nucleic Acid Res,1993,21(18):4 272-4280.
70.Benito E P,Prins T,Kan J A L.Application of differential display RT-PCR to the ananly-sis of gene expression in a plant-fungus interaction.Plant Mol Biol,1996,32:947-957.
71.Bleecker,A.B.and Kende,H..Ethylene:agaseous signal molecule in plants.Annu.Rev.Cell Dev.Biol.2000,16,1-18.
72.Carrera E,Bou J,Garcia-Martinez J L.Changes in GA 20-oxidase gene expression strongly affect stem length,tuber induction and tuber yield of potato plant.Plant J,2000,22:247-256.
73.Chen X F,W ang B Y,W u R.The level of a mRNA with sequence similarity to the old yellow enzyme-NADPH dehydro2genases increases in chenopodium rubrum cells in response to cytok inin.Jour Plant Physiol,1996,149(12):233-236.
74.DIACHENKO L B,LEDESMA J,CHENCHIKA A.Combing the Techniques of RNA Fingerp rinting and Differential Disp lay to Obtain Differentially Expressed mRNA.Biochem Biophys Res Commun,1996,219:824-828
75.Doss R P.Differential display without radioactivity-a modified procedure.BioTechniques,1996, 21 :410-412
76. Esther V D K, Kende H. Identification of a gibberellin induced gene in deepwater rice using differential display of mRNA. Plant Mol, 1995, 28: 589-592
77. Fang G, Hammar S, Grumet R. A quick and in expensive method for removing polysaccharides from plant genomic DNA. Bio Techniques, 1992, 13:52-55
78. FischerA, Saedler H, Theissen G. Restriction FragmentLength Polymorphism Coupled Domain Directed DifferentialDisplay: A Highly Efficient Technique for Expression Analysisof Multigene Families.Proceedings of the National Academy of Sciences of the United States of America, 1995, 92:5331-5335
79. Fujino K, Koda Y, Kikuta Y. Reorientation of cortical microtubules in the subapical region during tuberization in single-node stem segments of potato in culture. Plant Cell Physiol. 1995, 36: 891-895
80. Gang An, GuiZhen Luo, RobertW. Sensitive, Nonradioactive Differential Display Method Using Chemiluminescent Detection.Biology Techniques, 1996,20: 342 - 346
81. Hedden P. Regulation of gibberellin biosynthesis. Hooykaas P J J, Hall M A, Libbenga K R eds. Biochemistry and molecular biology of plant hormones. Elsevier Science: Amsterdam, 1999, 161-188
82. HUBANKM, SCHATZD G Identifying Difference in mRNA Expression by Reresentational Difference Analysis of cDNA. Nucl acids Res, 1994, 22 (25 ): 5 640 -5 648.
83. Jackson S D. Multiple signaling pathways control tuber induction in potato. Plant Physiol, 1999, 119:1-8
84. Jacobs A. Mo lecular analysis of cold resistance in ch ilean po tato species. Aminor field study. Work ing paper international rural development centre, Swedish U niversity of A griculture. Science, 1996, 305: 15-19
85. Kammer H V D , C Albrecht ,C Albrecht. Identification of genes regulated by muscarinic acetylcholine receptors: pplication of an improved and statistically comprehensive mRNA differential display technique. Nucleic Acids Research ,1999 , 27 (10) :2211-2218
86. Knaap E van der, Knende H. Identification of a gibberellin-induced gene in deepwater rice using differential-display of mRNA. PlantMo .Bio ., 1995, 28: 589- 592.
87. Koda Y. Possible involvement of jasmonates in various morphogenic events. Phsiol Plant, 1999, 100:639-646
88. Kolomiets M V, Hannapel D J, Chen H, Tymeson M, Gladon R J. Lipoxygenase Is Involved in the Control of Potato Tuber Development. The Plant Cell, 2001,13: 613-626
89. Lee S W, Tomasetto C , Sager R. Positive selection of candidate tumorsuppressor Genes by subtractive hybridization. Proc Natl Acad Sci USA, 1991 , 88 :2825 -2829
90. Leonard S. Jefferson Saot R. Kimball. Amino Acid Regulation of Gene Expression. Journal of Nutrition.2001,31:2460-2466
91. Liang P, Pardee A B. Differential Display of eukaryotic messenger RNA by means of the polymerase chain reaction. Science, 1992 , 257 (14): 961-971
92. LISITSYN N, W IGKER M. Cloning the Differences Betweentwo Complex Genomes.Science, 1993, 259 (5097) :946 - 951
93. Liu J, Xie C H, Liao Y, Huang D E. Effects of endogenous gibberellic acid and abscisic acid on the induction of potato microtubers in vitro. In: Liu Q and Kokubu J ads.Proceedings of 1st Chinese-Japanese Symposium on Sweetpotato and Potato. Beijing agricultural University, Beijing, 1995,267-275
94. Lohmann J, Schickle H, Bo sch TCG. REN display, a rapid and efficient method for nonradioactive differential display and mRNA isolation. Bio Tech, 1995; 18:200-202
95. Matz M and Lukyanov S A .Different strategies of differential display: areas of application. Nucleic Acids Res. 1998, 26(24):5537-5543
96. Mikhail Matz, Natalia Usman, Dmitryi Shagin.Ordered differential display: a simple method for systematic comparison of gene exp ression profiles.Nucleic Acids Research, 1997, 25: 2541 -2542.
97. Opsahl F H C , Dewnff E L, Dumas C. A novel endosperm-specific gene express -ed in a restricted region around the maize embryo. Plant Jour, 1997,12(1): 235 -246
98. Peng Liang, ArthurB Pardee.Differential Display of Eukaryotic Messenger RNA by Means of the Ploymerase Chain Reaction.Science, 1992, 257: 967 - 971
99. Radmacher W. Growth retardants: effects on gibberellin biosynthesis and other metabolic pathways. Annusl Review of Plant Physiology and Plant Molecular and biology,2000, 51:501-503
100. Robert P Doss.Differential Display Without Radioactivity A Modified Procedur.Biology Techniques, 1996, 21:408-412
101. Rossi Jaume A, Corral M, Tiai) R. Possible involvement of jasmonic acid in bulb formation of garlic (Allium satiuanl L). Acta Hort, 1997, 433:381-388
102. Sharma Y K, Davis K R. Iso lation of a novel A rabidop sis ozone2induced cDNA by differential display. Plant Mo.Bio., 1995, 29: 91- 98.
103. Simon HG, Oppenheimer S. A dvanced mRNA differential disp lay:isolation of a new differential regulated myosin heavy chain-ncoding gene in amphibian limb regeneration. Gene, 1996, 72: 175-181