黄瓜子叶扩张与细胞分裂素的关系

BA(benzyl adenine)专一性地促进离体黄瓜子叶的扩张。为了研究BA的作用机理,我们采用间接EIISA和HPLC的方法测定了子叶扩张过程中内源玉米素 (Z) 和玉米素核苷(ZR)含量的变化。离体黄瓜(Cucumis sativus,津研4号)子叶用100mg／1的BA培养,72小时之后,处理子叶鲜重的增加比对照高70％。Z+ZR在BA处理的子叶中有大量的积累。结果表明BA可能诱发了黄瓜子叶中的细胞分裂素生物合成和代谢的某些基因。     

细胞分裂素和钾离子对离体黄瓜子叶扩张和代谢变化的比较研究

10 ppm的BA(相当于0.044mM)在暗中强烈地促进离体黄瓜子叶扩张、类胡萝卜素积累及核酸合成,相同条件下,单用KCl(75mM)则没有作用或作用很小。但在光下,KCl明显地促进子叶扩张、核酸合成和叶绿素积累,BA在光下对前两种反应比KCl强得多。看来钾离子的单独作用与光有密切关系,而BA的作用在光下仅略大于暗中。KCl与BA引起子叶扩张的形态变化及对叶绿素合成的影响也是不同的。无论是在暗中或是光下,KCl与BA合用,对离体黄瓜子叶扩张、类胡萝素或叶绿素积累及核酸合成,均有明显的增效作用。实验表明:BA和钾离子在调节黄瓜子叶扩张及代谢变化中的关系十分密切,但它们的作用方式却不相同。     

1,4—二取代酰氨基硫脲类化合物(DATCDs)的植物生理效应与应用研究——Ⅴ.DATCD-A对离体黄瓜子叶扩张及核酸代谢的影响

本实验以离体黄瓜子叶为材料,研究了 DATCD—A 对子叶扩张及核酸代谢的影响。结果表明,DATCD—A 可显著地促进离体黄化子叶扩张,使其鲜重明显增加;并且在处理后期逐渐提高子叶干重。在离体子叶扩张变绿的过程中,DATCD—A 促进离体黄化子叶 RNA、DNA 含量和 DNA/RNA 比率明显上升,且 RNA 的增加发生在 DNA 合成增加之前。凝胶电泳证明,RNA 的增加主要是25s 和18s 的 rRNA。     

云南山楂中吲哚乙酸、脱落酸、玉米素和玉米素核苷的内源水平及其生根率

以云南山楂(Crataegus scabrifolia (Franch.)Rchd.)几个株系的成年树和实生苗的离体培养芽条为材料,用酶联免疫吸附测定法(ELISA)测定了它们的叶片中吲哚乙酸(IAA),脱落酸(ABA),玉米素和玉米素核苷(Z+ZR)的内源水平,探讨了这三种激素及其之间的平衡与生根率的相关性。结果发现:1.IAA内源水平之差异存在于株系间。2.内源(Z+ZR)在株系间无显著差异,而幼树发育至成年树的过程中,与IAA的平衡是多样的,同成年树的生根率似有以下相关性,当IAA水平降低,同时(Z+ZR)水平升高,有利于生根,反之则抑制生根。3.ABA内源水平无论成年或幼年树都呈低水平且无显著差异。     

细胞分裂素对黄瓜子叶张开及其下胚轴倒钩伸直的影响

6-苄基嘌呤(BA),玉米素和激动素引起黄瓜(cucumsis sativus,品种:津研4号)子叶的张开。其它的植物生长调节剂,如赤霉酸(GA_3),吲乙酸(IAA),乙烯,脱落酸(ABA),以及有关化合物,如 KCl 和腺嘌呤均无此作用。BA 的浓度在0.01ppm 时也能产生这一效应。因此这一作用能作细胞分裂素生物鉴定的定性方法。KCl 对细胞分裂素诱导的子叶张开效应有增效的作用。在去根下胚轴倒钩基部供给 BA(10ppm)能促进留子叶倒钩的伸直,光照能促进这一效应。然而,BA 却抑制不留子叶的倒钩的伸直,特别在光下更明显。子叶的存在对倒钩维持弯曲是有利的。     

苦参碱对植物生长的刺激作用

应用植物激素生物试法测定了苦参碱（matrine)的某些植物生长调节活性,实验结果表明：苦参碱明显促进了离体黄瓜子叶鲜重和干重的增加;苦参碱降低了光下（5000lx,26℃）培养的离体黄瓜子叶的总叶绿素含量,但促进了离体黄瓜子叶的光合作用;苦参碱还显著促进了在暗中（25℃）培养的离体黄瓜子叶的生根作用和根的生长,这些现象说明苦参碱具有某些明显的植物生长刺激活性。     

多胺对离体黄瓜子叶花芽构建的影响

研究了外源多胺对离体黄瓜(CucumissativusL.)子叶内在多胺含量和花芽构建的影响。在MS培养基中添加5×10-2mmol·L-1亚精胺(Spd)、精胺(Spm)能明显提高子叶内Spd含量和SpdPut值,子叶成花率也明显高于对照组,而添加5×10-2mmol·L-1Put、5mmol·L-1环己胺硫酸盐(CHAS)培养的结果则相反。表明子叶内Spd含量和SpdPut与成花率呈正相关关系。在添加5mmol·L-1D精氨酸(DArg)处理后,子叶内SpdPut高于Spd处理,但子叶成花率低于对照组,表明Put、Spd含量也影响离体子叶花芽的构建。在实验中添加CHAS处理后子叶内Spm含量高出对照组10倍,子叶成花率却为0,其它各处理与对照组的内源Spm变化基本一致,成花率却差别明显,表明子叶内Spm含量与黄瓜子叶花芽构建之间无明显相关性。     

苦参碱对植物生长的刺激作用

应用植物激素生物试法测定了苦参碱（matrine)的某些植物生长调节活性,实验结果表明：苦参碱明显促进了离体黄瓜子叶鲜重和干重的增加;苦参碱降低了光下（5000lx,26℃）培养的离体黄瓜子叶的总叶绿素含量,但促进了离体黄瓜子叶的光合作用;苦参碱还显著促进了在暗中（25℃）培养的离体黄瓜子叶的生根作用和根的生长,这些现象说明苦参碱具有某些明显的植物生长刺激活性。     

金雀花碱的植物生长调节活性

应用植物激素生物试法测定了金雀花碱（cytisine)的某些植物生长调节活性,实验结果表明：金雀花碱抑制小麦芽鞘切段的伸长,但促进黑暗中（26℃）培养的离体黄瓜子叶鲜重的增加;能明显促进黑暗中（25℃）培养的离体黄瓜子叶的生根作用和根的生长。能显著提高离体黄瓜子叶中吲哚乙酸氧化酶的活性,这些现象说明,金雀花碱具有明显的植物生长调节活性。     

金雀花碱的植物生长调节活性

应用植物激素生物试法测定了金雀花碱（cytisine)的某些植物生长调节活性,实验结果表明：金雀花碱抑制小麦芽鞘切段的伸长,但促进黑暗中（26℃）培养的离体黄瓜子叶鲜重的增加;能明显促进黑暗中（25℃）培养的离体黄瓜子叶的生根作用和根的生长。能显著提高离体黄瓜子叶中吲哚乙酸氧化酶的活性,这些现象说明,金雀花碱具有明显的植物生长调节活性。     

Involvement of cytokinins in the germination of chick-pea seeds

Eight cytokinins detected in germinated chick-pea (Cicer arietinum L. var. Castellana) seeds were first present in the embryonic axes but appeared in the cotyledons after 12h of germination. The cytokinins detected in the cotyledons originate in the embryonic axes, but no passage of these substances from the cotyledons to the axes was detected, except when the seeds were treated with red light.It is concluded that the role played by the embryonic axis in mobilizating the main reserves of the cotyledons is mainly effected through these cytokinins. Both natural and synthetic cytokinins exert an important regulatory role in the hydrolysis of reserve proteins and calcium could be involved as an intermediate.Abbreviations BA benzyladenine - cot. cotyledon - (diH)Z dihydrozeatin - (diH)ZR dihydrozeatin riboside - GZR glycosyl zeatin riboside - 2iP 277-1 - iPA 277-2 riboside - Kin kinetin - Z zeatin - ZG zeatin glucoside - ZR zeatin riboside     

Cytokinins in maturing and germinatingLupinus luteus L. seeds

³H-labelled zeatin riboside (ZR) was applied to pod walls of intactLupinus luteus L. plants. Metabolites present in mature, dry seeds were zeatin nucleotide (ZNT), zeatin riboside (ZR) and zeatin (Z), zeatin O-glucosides and lupinic acid (LA), and the corresponding dihydro-derivatives of the cytokinins listed. Endogenous cytokinins were rapidly metabolised in germinating seeds. In seeds labelled with [³H]ZR for 90 min following a 2 h period of imbibition in water, ZR was actively converted to ZNT and dihydro-ZNT but the prevailing CTK was Z in cotyledons and ZR in embryo axes (EA); later LA and dihydro-LA, and O-glucoside metabolites accumulated. When [³H] zeatin was introduced into imbibing seeds, it was converted to dihydro-ZNT, ZNT, dihydro-ZR, ZR and dihydro-Z; in EA of the Z-labelled seeds, dihydro-ZR and ZR were the main cytokinins. After incubation of the Z-labelled seeds for 6 h in water, the ratios of dihydro-ZNT: ZNT and dihydro-ZR: ZR were, respectively, 20: 1 and 3.4: 1 in EA, and 3.5: 1 and 1.4: 1 in cotyledons.     

Induction of chitinase activity by exogenous cytokinins in excised dark-grown cucumber cotyledons: involvement of Ca and staurosporine-sensitive protein kinase(s) in cytokinin signaling

Cotyledons were excised from 7-day-old dark-grown cucumber seedlings and treated with water, benzyladenine (BA), kinetin (K), zeatin (Z), or zeatin riboside (ZR) in dark after endogenous cytokinin depletion. We have compared changes in chitinase (EC. 3.2.1.14) activity induced by these cytokinins. We find that the activities of chitinase and its isoforms increase by approximately 3- to 6-fold following BA, Z, and ZR treatments. Among these treatments, Z was more effective. K was totally ineffective in inducing chitinase activity. Immunoblot analysis suggests that the cytokinin Z-induction of enzyme activity is due to the induction of higher chitinase protein levels and not the activation of existing enzyme. Furthermore, the Z-induced chitinase activity and its protein accumulation were completely inhibited by the protein kinase inhibitor staurosporine, whereas the protein phosphatase inhibitor sodium fluoride was not effective in such inhibitions. Treatment of cotyledons with extemal CaCl₂ and calcium ionophore increased the basal chitinase activity by 6- and 5-fold, respectively. Moreover, the effects of staurosporine, sodium fluoride, and Ca²⁺ on Z-induced chitinase activity correlate with their effects on chitinase protein levels. Taken together, our data suggests Ca²⁺ and staurosporine-sensitive protein kinase(s) as components of the cytokinin transduction machinery involving induction of chitinase in cucumber.     

Further studies on the effects of different auxins and cytokinins on flower formation in thin cell layers of Nicotiana tabacum: The effects of zeatin riboside, dihydrozeatin and dihydrozeatin riboside

Organogenesis in thin cell layers of Nicotiana tabacum L. was studied in relation to the effects of natural and synthetic auxins in combination with various cytokinins. All cytokinins tested, benzyladenine (BA), kinetin, zeatin (Z), zeatin riboside (ZR), N⁶-Δ²-isopentenyl) adenine (IPA), dihydrozeatin [(diH)Z] and dihydrozeatin riboside [(diH)ZR], seem to be active in flower bud formation. In addition to the initiation of flower buds, vegetative buds or roots were also formed on the explants in the presence of BA, Z or IPA as exogenous cytokinins. Only dihydrozeatin and its riboside stimulated the initation of flower buds alone (as is known for kinetin), especially if supplemented with indole-3-acetic acid (IAA) as exogenous auxin. A high number of explants with flower buds was also found with high cytokinin/2,4-D ratios. In these conditions the presence of (diH)Z yielded the higest number of flower buds per explant.     

Direct and efficient plant regeneration from leaf explants of Solanum tuberosum l. cv. Bintje

Summary A two-step procedure was used for plant regeneration from in vitro grown leaf strips (2–3 mm wide) of cv. Bintje. Step I medium was designed with 2,4-dichlorophenoxycetic acid (2,4-D) at 0.0 or 9.0 M, in combination with 2.28 M kinetin (K), benzyl adenine (BA), zeatin (Z) or zeatin riboside (ZR). Step II media were 2,4-D-free media containing 5.78 M gibberellic acid (GA3) and growth regulators similar to those of step I media. Leaf explants cultured in medium I containing zeatin riboside or zeatin for 6 days and then subcultured in medium II containing zeatin riboside produced numerous shoots without callus formation. Zeatin riboside containing step I and II media caused shoot regeneration in a high number (97.5±2.2) of explants. Approximately, 33.7±8.4 shoots were regenerated from each leaf explant.Abbreviations BA benzyladenine - Z zeatin - ZR zeatin riboside (trans isomer) - 2,4-D 2,4-dichlorophenoxyacetic acid     

The stability and biological activity of cytokinin metabolites in soybean callus tissue

The activity, uptake and metabolism of cytokinin metabolites was determined in soybean (Glycine max (L.) Merr.) callus tissue. The following activity sequence was established: zeatin riboside (ZR)>zeatin (Z)>O-glucosides of Z, ZR and their dihydro derivatives>lupinic acid (an alanine conjugate of Z)>7- and 9-glucosides of Z which were almost inactive. The 7- and 9-glucosides and lupinic acid were taken up very slowly by the callus tissue and showed great metabolic stability, but some degradation to 7-glucosyladenine, 9-glucosyladenine and the 9-alanine conjugate of adenine occurred. Compared with its aglycone, O-glucosyl-ZR exhibited slow uptake and greatly enhanced stability but gas chromatographic-mass spectrometric analysis showed that appreciable amounts were hydrolyzed to ZR in the tissue. Both ZR and O-glucosyl-ZR were metabolised extensively, with adenine, adenosine, and adenine nucleotide(s) as the major metabolites. A diversity of minor metabolites of ZR were identified, including O-glucosides, lupinic acid and dihydrolupinic acid. The metabolism of ZR was suppressed by 3-isobutyl-1-methylxanthine. When compared with the soybean callus line normally used for cytokinin bioassays (cv. Acme, cotyledonary callus), related callus lines exhibited greatly differing growth responses to cytokinin: however, these were not reflected in marked differences in metabolism.Abbreviations GC-MS gas chromatography-mass spectrometry - HPLC high-performance liquid chromatography - LA lupinic acid - OGZR O--D-glucopyranosylzeatin riboside - TLC thin-layer chromatography - IMX 3-isobutyl-1-methylxanthine - Z zeatin - ZR zeatin riboside     

Effect of applied and internal hormones on vitrification and apical necrosis of different plants cultured in vitro

Development of vitrification and apical necrosis was followed in Camellia sinensis, Gerbera jamesonii, Malus domestica and hybrid Populus tremula x P. alba shoots cultured in vitro on Murashige & Skoog (MS) medium with different concentrations of growth regulators. High humidity in the culture vessels and excess of BA in the medium were found to be the major factors influencing vitrification. Lack of exogenous cytokinin in the medium during successive subcultures induced apical necrosis in poor-rooting species (Malus domestica, Camellia sinensis). The level of internal phytohormones (ABA, IAA, IPA, 2iP, Z, ZR) was determined in the apple shoots by means of ELISA. The content of internal cytokinins in the vitrified apple shoots was several times greater than in normal ones, which supports the hypothesis that excess of cytokinins, inducing rapid divisions of cells in meristems in the atmosphere with high humidity, is responsible for vitrification. Apical necrosis of the plantlets that appeared after cultivation on cytokinin-free medium is the result of deficiency in endogenous hormones in apple shoots and this being confirmed by analysis of endogenous hormones in apple shoots.Abbreviations BA benzyladenine - BHT butylated hydroxy-toluene - ABA abscisic acid - IAA indole-3-acetic acid - ELISA enzyme-linked immunosorbent assay - IPA isopentenyladenosine - 2iP isopentenyladenine - NAA naphthyl-3-acetic acid - TBS trishydroxymethylaminomethane buffered saline - TLC thin layer chromatography - Z zeatin - ZR zeatin riboside     

小麦与玉米及鸭茅状摩擦禾杂交后胚发育过程中内源激素的动态变化

小麦（Ｔｒｉｔｉｃｕｍ ａｅｓｔｉｖｕｍ Ｌ．）与玉米（Ｚｅａ ｍ ａｙｓＬ．）及鸭茅状摩擦禾（Ｔｒｉｐｓａｃｕｍ ｄａｃｔｙｌｏｉｄｅｓ Ｌ．）杂交后,由于胚乳组织不能正常形成,从而导致胚很难在母本小麦植株上发育至成熟。通过对杂交后的胚不同发育时期的子房内源ＩＡＡ、玉米素（Ｚ）及玉米素核苷（ＺＲ）和ＡＢＡ 的ＥＬＩＳＡ 分析,发现远缘杂交后的胚发育过程中子房内源ＩＡＡ 的浓度明显低于正常发育的子房;而且Ｚ＋ ＺＲ 和ＡＢＡ 的含量的动态变化也和正常自交发育的子房不同,然而ＩＡＡ／（Ｚ＋ ＺＲ）和ＩＡＡ／ＡＢＡ 的比率变化在远缘杂交与正常自交的子房之间有某些相似趋势。ＩＡＡ、Ｚ＋ ＺＲ和ＡＢＡ 与这３ 种激素之和的比率变化在上述两类子房中也有相似之处。可以推断,植物胚胎的正常发育需要多种激素的动态平衡及某一时期某一激素的主导性作用。远缘杂交后的胚之所以不能够顺利发育至成熟,除了缺乏胚乳组织的营养保护之外,或许也和其内源激素的水平及多激素平衡体系偏离常规动态变化有关联     

Benzyladenine and derivatives-their significance and interconversion in plants

Recently benzyladenine has been isolated as a natural cytokinin from a number of plants. The natural occurrence of this cytokinin will change the attitude with which physiologists view this hormone. This review attempts to put into context what is known about this cytokinin and its derivatives and to compare and contrast its metabolism and the function and physiological action of its various metabolites. Nothing is known about the biosynthesis of benzyladenine. Its structure would suggest that its biosynthetic pathway may differ considerably from that of zeatin and iso-pentenyladenine.Abbreviations Ade adenine - Ado adenosine - BA benzyladenine - [9R]BA BA ribonucleoside - [9R-MP]BA BA nucleotide - [9R-DP]BA BA dinucleotide - [9R-TP]BA BA trinucleotide - [3G]BA BA 3 glucoside - [7G]BA BA 7 glucoside - [9G]BA BA 9 glucoside - [9R-G]BA BA 9-ribosylglucoside - [9Ala]BA BA alanine-conjugate - (2OH)BA BA ortho-OH - (2OH)[9R]BA BA ortho-Oh-riboside - KN kinetin - [9R]KN KN ribonucleoside - DHZ dihydrozeatin - Z trans-zeatin - [9R]Z zeatin ribonucleoside - [7G]Z zeatin-7-glucoside - [9G]Z zeatin-9-glucoside - [9Ala]Z zeatin alanine-conjugate - (OG)[9R]Z O-glucoside of zeatin ribonucleoside - [9R-MP]Z zeatin nucleotide - iP iso-pentenyladenine - [9R]iP iP ribonucleoside