植物体内的芳环细胞分裂素

芳环细胞分裂素包括6-苄基氨基嘌呤（BA）,6-（3-羟苄基氨基）-嘌呤（mT）,6-（2-羟苄基氨基）-嘌呤(oT)和它们的衍生物。它的代谢具有相互转换、羟基化作用、结合作用和氧化降解四个特征。本文从信号的感受,转导和应答阐明细胞分裂素在细胞和分子水平上的 功能。     

细胞分裂素促进小麦硝酸还原酶的诱导

细胞分裂素促进硝酸还原酶的诱导,脱落酸强烈抑制该酶的诱导,并抵消细胞分裂素的作用。6-苄基腺瞟呤(6-BA)的促进效应与 NO_3~-的诱导作用有关。无 NO_3~-存在时,6-BA 无直接诱导作用,适宜浓度的 NO_3~-可诱导较高的酶活性,这时6-BA 的促进作用也较强。无光照时,NO_3~-只能诱导黄化叶片产生很低的酶活性,这时6-BA 的促进作用也很弱。在1—2小时的诱导期内,环己酰亚胺抑制酶的诱导。结果表明细胞分裂素对酶诱导的促进作用不仅与 NO_3~-的存在有关,还与 NO_3~-诱导硝酸还原酶的必要条件有关,即依赖于酶的诱导过程。     

苦参组培快繁技术体系的初步研究

采用正交实验设计(L934)用MS、White和改良MS培养基,分别加入不同质量浓度的玉米素(ZT)、6-苄基氨基腺嘌呤(6-BA)、异戊烯基腺嘌呤(2-ip)进行苦参组培快繁技术体系研究。结果表明,以苦参种子萌发的幼苗为外植体效果较好。3种基本培养基对诱导芽增殖效果差异不显著;在基本培养基中分别加入6-BA、2-ip和ZT诱导芽增殖,但3种细胞分裂素之间差异不显著。培养基中含2 mg/L 6-BA(或ZT或2ip)增殖效果最好,其次是3mg/L的细胞分裂素;以改良MS 2 mg/L 6-BA是苦参较好的增殖培养基;组培苗在1/2 MS 1.5 mg/L IBA培养基上根诱导效果最好(98%),移栽到珍珠岩 蛭石基质(1∶1)中成活率最高(96%)。     

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。     

伤流液中玉米素的高效液相色谱测定

在植物体内,天然存在的细胞分裂素(Cytokinin)主要是玉米素和与玉米素化学结构相似的化合物6-(r.r-二甲基烯丙基氨基)—嘌呤。尤以玉米素活性最强,含量最多。其活性大约是细胞分裂     

细胞分裂素及其应用

细胞分裂素是目前人们已知道的五大类植物激素(即生长素、赤霉素、细胞分裂素、脱落酸和乙烯)之一,它具有腺嘌呤环结构。这类物质的共同特点是在腺嘌呤环的第六位置氨基上有特定的取代物,对细胞分裂及分化等有重要作用。细胞分裂素在植物体内多分布在细胞分裂旺盛的组织和器官中,如根尖、茎尖、未成熟的种子、萌发的种子以及正在发育的果实等。实验证明,根尖是合成细胞分裂素的场所。在高等植物中已发现的细胞分裂素有16种,其中最常见的是玉米素、玉米素核苷、二氢玉米素和异戊烯基腺苷     

天麻的化学研究(Ⅳ)几种国产天麻属植物的化学成分

我们曾报道从云南昭通产药用天麻(Gastrodia elata Bl.)及其鲜品中分离到九种酚性成分,即:天麻素(1),对羟基苯甲醇(2),对羟基苯甲醛(3),3.4-二羟基苯甲醛(4),4,4′-二羟基二苯基甲烷(5),4,4′-二羟基二苄醚(6),对羟苄基乙基醚(7),4-乙氧甲苯基-4′-羟苄基醚(8),以及三〔4-(β-D-葡萄吡喃糖基)-氧-苄基〕-柠檬酸酯(即Parishin)(9)。并对其中镇静催眠的有效成分(1)进行了合成。本文进一步比较不同产地药用天麻及各种国产天麻属植物的化学成分。     

拟诺卡氏放线菌YIM90087的代谢产物研究

对拟诺卡氏菌YIM90087的固体发酵提取物进行了化学成分研究,从中分离得到10个化合物。通过波谱数据分析,鉴定其结构分别为:(3S)-3-苄基-2,5-哌嗪二酮(1)、(3S,6S)-3-(2-丁基)-6-(1-羟乙基)-2,5-哌嗪二酮(2)、(3S,6S)-3-(2-丁基)-6-羟甲基-2,5-哌嗪二酮(3)、(4S)-4-苄基-1,3-恶唑-2,5-二酮(4)、6'-羟基-4,2',3',4'-四甲氧基-对三联苯(5)、(3S,6S)-3-(4-羟苄基)-6-异丁基-2,5-哌嗪二酮(6)、(3R,8aS)-3-异丙基-六氢吡咯并[1,2-a]吡嗪-1,4-二酮(7)、(3S,6S)-3-异丁基-6-甲基-2,5-哌嗪二酮(8)、(3S,6S)-3-苄基-6-(1-羟乙基)-2,5-哌嗪二酮(9)和(3S,6S)-3-苄基-6-(1-羟丙基)-2,5-哌嗪二酮(10)。化合物4目前只见化学合成报道,为新天然产物。     

拟诺卡氏放线菌YIM90087的代谢产物研究北大核心CSCD

对拟诺卡氏菌YIM90087的固体发酵提取物进行了化学成分研究,从中分离得到10个化合物。通过波谱数据分析,鉴定其结构分别为:(3S)-3-苄基-2,5-哌嗪二酮(1)、(3S,6S)-3-(2-丁基)-6-(1-羟乙基)-2,5-哌嗪二酮(2)、(3S,6S)-3-(2-丁基)-6-羟甲基-2,5-哌嗪二酮(3)、(4S)-4-苄基-1,3-恶唑-2,5-二酮(4)、6'-羟基-4,2',3',4'-四甲氧基-对三联苯(5)、(3S,6S)-3-(4-羟苄基)-6-异丁基-2,5-哌嗪二酮(6)、(3R,8aS)-3-异丙基-六氢吡咯并[1,2-a]吡嗪-1,4-二酮(7)、(3S,6S)-3-异丁基-6-甲基-2,5-哌嗪二酮(8)、(3S,6S)-3-苄基-6-(1-羟乙基)-2,5-哌嗪二酮(9)和(3S,6S)-3-苄基-6-(1-羟丙基)-2,5-哌嗪二酮(10)。化合物4目前只见化学合成报道,为新天然产物。     

棉花子房中存在细胞分裂素的专一结合蛋白

棉花子房细胞提取物1000r/min沉淀组分(P_1)和10 000r/min沉淀组分(P_(10))均能与6-BA专一结合,但不能与ABA专一结合。非标记的激动素、玉米素和6-BA等细胞分裂素类物质均可将与P_1结合的[~3H]6-BA取代下来,其它激素IAA和ABA、细胞分裂素的结构类似物cAMP和腺嘌呤均无取代作用。胰蛋白酶处理可明显降低P_1和6-BA的专一结合,说明在棉花子房中存在细胞分裂素的专一结合蛋白。二硫苏糖醇(DTT)、半胱氨酸和还原型谷胱甘肽强烈抑制P_1的专一结合,提示某种可还原基团,很可能是二硫键,定位于结合蛋白,且与其结合活性有关。在子房细胞里存在能钝化细胞分裂素结合蛋白活性的物质,其钝化能力是热不稳定的。     

The aromatic cytokinins

After the discovery of kinetin (Miller et al. 1956, J. Am. Chem. Soc. 78: 1345–1350) there was a flurry of syntheses that led to the finding of 6-benzylaminopurine (BA), an active and easily obtainable cytokinin. Much research into cytokinin physiology was subsequently done with this substance. Further, the isolation and unequivocal identification of natural BA and the high biological activity of its meta -hydroxylated analogues stimulated the search for other natural aromatic cytokinins. Screening was accomplished by ELISA of HPLC fractions using antisera against ortho - and meta -hydroxybenzyladenosine. Subsequent isolation and decisive identification by mass spectrometry led to discovery of a broad spectrum of endogenous plant growth substances structurally similar to a highly active compound, meta -topolin (6-[3-hydroxybenzyl-amino]purine), and to its less active analogue, ortho -topolin (6-[2-hydroxybenzyl-amino]purine). The structures of such aromatic cytokinins suggest considerably different biosynthetic pathways from that of zeatin and related isoprenoid cytokinins. From a physiological viewpoint, aromatic cytokinin metabolism can be classified under four main headings analogous to isoprenoid cytokinins: interconversion, hydroxylation, conjugation, and oxidative degradation. This review attempts to put into context what is known about 9-alkyl-BAs and compares their metabolism in regard to the practical use of cytokinins in agriculture and biotechnology. The recently discovered unusual specificity of additionally C2,N9-disubstituted aromatic cytokinins toward cell cycle kinases, suggests that these cytokinin-derived growth regulators may selectively inhibit certain steps of the cell cycle. The functional overlap of the aromatic cytokinins with those of their isoprenoid counterparts and cytokinin inhibitors, in relation to growth and developmental processes in plants, has yet to be determined.     

Immunodetection and Identification of N-(o-Hydroxybenzylamino)Purine as a Naturally Occurring Cytokinin in Populus x canadensis Moench cv Robusta Leaves

A highly specific and sensitive enzyme-linked immunosorbent assay (ELISA) was developed for 9-β-d-ribofuranosyl-N⁶-(o-hydroxybenzylamino)purine [(oOH)[9R]BAP] and structurally related cytokinins. As little as 3 femtomoles of the compound could be detected by this method. Cross-reactivity studies demonstrated the specificity of four polyclonal antibodies for (oOH)[9R] BAP and its free base in preference to a range of natural cytokinins and other purines. After evaluating the method by internal standardization employing [2-³H](oOH)[9R]BAP of high specific radioactivity as recovery marker by dilution analyses and by immunohistograms, it was possible to apply ELISA to quantify (oOH)[9R]BAP in plant extracts. In addition to (oOH)[9R]BAP, an unknown cytokinin reacting with the same antibody was detected in high performance liquid chromatography-fractionated extracts of mature Populus × canadensis Moench cv Robusta. The structure of the new compound was determined by gas chromatography-mass spectrometry and finally confirmed by synthesis as N⁶-(o-hydroxybenzylamino)purine.     

Cytokinins from terminal buds of Euphoria longana during different growth stages

The presence of endogenous cytokinins were detected in the terminal buds of longan ( Euphoria longana Lam.) after purification by ion exchange and Sephadex LH-20 chromatography, and bioassay, enzymic degradation, high-performance liquid chromatography and gas chromatography-mass spectrometry. Permethylated derivatives of two highly active cytokinin glucoside compounds from dormant buds were: 6-(4-O-β-D-glucosyl-3-methyl-but-2-enylamino) purine (zeatin-O-glucoside) and 9-β-D-ribofuranosyl-6-(4-hydroxy-3-methyl-but-2-enylamino) purine (zeatin riboside-O-glucoside). Simultaneously, four active cytokinins from buds at the stages of leaf flush and flower bud initiation were identified as 6-(4-hydroxy-3-methyl-but- trans -2-enylamino) purine (zeatin), zeatin-9-β-D-ribofuranosylpurine (zeatin riboside), 6-(3-methyl-2-butenyl) aminopurine (isopentenyladenosine, 2iPA) and N-(3-methyl-2-butenyl) adenine (isopentenyladenine, 2iP). The total cytokinin levels were low at leaf flush, with the zeatin and zeatin riboside in the buds about 70% of the total. In the transition of the terminal bud from leaf flush to dormancy, the principal cytokinins were zeatin-O-glucoside and zeatin riboside-O-glucoside. However, significant decreases in the levels of zeatin-O-glucoside and zeatin riboside-O-glucoside and increases in those of zeatin, zeatin riboside, 2iPA and 2iP were observed at flower bud initiation. It is suggested that in longan, the cytokinins that are translocated to the shoots are accumulated in the buds at the dormant stage, and that later there is a considerable increase in free cytokinins during flower bud initiation, leading to the promotion of flower bud development.     

Anticytokinin activity of 4-substituted triazolo[4,5-d]pyrimidines and 4-substituted pyrazolo[3,4-d]pyrimidines

The synthesis and physiological activity of some novel 4-substituted triazolo[4,5-d]pyrimidines and 4-substituted pyrazolo[3,4-d]pyrimidines are described. Most of the compounds possessed high anticytokinin activity towards purine (benzyladenine) and phenylurea (4-PU-30) type cytokinins. 1-Benzyl-4-ethoxycarbonylpiperazinyl-1H-1,2,3-triazolo[4,5-d]pyrimidine almost completely removed cytokinin stimulated effects—betacyanin synthesis in Amaranthus caudatus cotyledons; growth of radish cotyledons and retention of chlorophyll in leaf explants. Some chemical structurephysiological activity relationships have been established.     

Cytokinins induce direc somatic embryogenesis of <Emphasis Type="Italic">Dendrobium</Emphasis> chiengmai pink and subsequent plant regeneration

Summary Four auxins (2,4-dichlorophenoxyacetic acid [2,4-D], indole-3-acetic acid [IAA], indole-3-butyric acid [IBA], and naphthaleneacetic acid [NAA]), and five cytokinins (N ⁶-[2-isopentenyl]-adenine [2iP], N ⁶-benzyladenine [BA], 6-furfurylaminopurine [kinetin], 1-phenyl-3-(1,2,3-thiadiazol-5-yl)-urea [TDZ], and 6-[4-hydroxy-3-methylbut-2-enylamino]purine [zeatin]) were examined for their effects on direct embryo induction from leaf explants of Dendrobium cv. Chiengmai Pink cultured on 1/2 Murashige and Skoog (MS) medium. Whether in light or darkness, explants easily became necrotic and no embryos were obtained on growth regulator-free or auxin-containing media after 60 d of culture. By contrast, five cytokinins tested induced direct embryo formation from leaf explants, and explants cultured in light had a higher embryogenic response compared with those cultured in darkness. The best condition for direct embryo induction was at 18.16 μM TDZ cultured in light for 60 d, where 33% of explants formed a mean number of 33.6 embryos per explant. During subculture on growth regulator-free 1/2 MS medium, embryos gradually developed into plantlets. Secondary embryogenesis was occasionally found on sheath leaves of embryos. Regenerated plantlets were successfully transplanted and grown in a greenhouse environment.     

Cytokinin-induced differentiation of human myeloid leukemia HL-60 cells is associated with the formation of nucleotides, but not with incorporation into DNA or RNA

Cytokinins are important purine derivatives that act as hormones to control many processes in plants. Cytokinins such as isopentenyladenine (IPA), kinetin and benzyladenine were very effective at inducing the granulocytic differentiation of human myeloid leukemia HL-60 cells. The metabolism of cytokinins to their nucleotides was closely associated with cytokinin-induced differentiation and growth inhibition. When the cells were incubated with [14C]-benzyladenine, radioactivity was significantly incorporated into RNA and DNA. However, the radioactive nucleotides in RNA or DNA were adenine nucleotides, not benzyladenine nucleotides, suggesting that cytokinins were not incorporated into RNA and DNA. The benzyladenine nucleotides were not stably released into the medium in intact form. Cytokinins effectively induced a phosphorylated (active) form of mitogen-activated protein kinase (MAPK). MAPK activation was necessary for cytokinin-induced differentiation, because PD98059, an inhibitor of MAPK kinase, suppressed the differentiation induced by cytokinins. These results suggest that cytokinin nucleotides themselves play an important role in inducing the differentiation of HL-60 cells.     

Mass Spectroscopic Identification of Cytokinins: Glucosyl Zeatin and Glucosyl Ribosylzeatin from Vinca rosea Crown Gall

Mass spectrographic and chemical studies of the permethyl and trimethylsilyl ethers of two new cytokinins isolated from Vinca rosea crown gall callus cultures by Peterson and Miller (Plant Physiol 59: 1026-1028) indicate that they are 6-(4-0-beta-d-glucopyranosyl-3-methyl-trans-but-2-enylamino) purine (glucosyl zeatin) and 9-beta-d-ribofuranosyl-6 (4-0-beta-d-glucopyranosyl-3-methyl-trans-but-2-enylamino) purine (glucosyl ribosylzeatin). The nature of the mass spectra of the permethylated cytokinins suggests that these derivatives may have considerable utility in the detection of low levels of cytokinins in plant material.     

Synthesis of 2-amino-6-(4-[11C]methoxyphenylthio)-9-[2-(phosphonomethoxy)ethyl]purine bis(2,2,2-trifluoroethyl) ester as a novel potential PET gene reporter probe for HBV and HSV-tk in cancers

Acyclic nucleoside 2-amino-6-(4-methoxyphenylthio)-9-[2-(phosphonomethoxy)ethyl]purine bis(2,2,2-trifluoroethyl) ester (ABE, 1) is a new hepatitis B virus (HBV) specific antiviral reagent and shows high anti-HBV activity. Carbon-11 labeled ABE may serve as a novel reporter probe for positron emission tomography (PET) to image HBV and herpes simplex virus thymidine kinase (HSV-tk) in cancers. The radiolabeling precursors 2-amino-6-(4-hydroxyphenylthio)-9-[2-(phosphonomethoxy)ethyl]purine bis(2,2,2-trifluoroethyl) ester (10) and 2-N-Boc protected analogue 2-N-bis(Boc)amino-6-(4-hydroxyphenylthio)-9-[2-(phosphonomethoxy)ethyl]purine bis(2,2,2-trifluoroethyl) ester (12), and the reference standard ABE were synthesized from bis(trifluoroethyl) (2-iodoethoxy)methylphosphonate (5), guanine (6), and 2-amino-6-chloropurine (8). The target radiotracer 2-amino-6-(4-[11C]methoxyphenylthio)-9-[2-(phosphonomethoxy)ethyl]purine bis(2,2,2-trifluoroethyl) ester ([11C]ABE, [11C]1) was prepared by O-[11C]methylation of the unprotected HO-precursor 10, or 2-N-Boc protected HO-precursor 12 with [11C]methyl triflate followed by a quick deprotection reaction, and isolated by solid-phase extraction (SPE) purification in 40-55% radiochemical yields.     

Antagonistic Effects of Triazolo[4,5-d]pyrimidine and Pyridylurea Derivatives on Cytokinin-Induced Cytokinin Oxidase/Dehydrogenase Activity in Young Pea Plants

The effect of strong and weak cytokinin antagonists, belonging to the groups of triazolo[4,5-d]pyrimidines (TP), and pyridyl-phenylurea derivatives (PU), on cytokinin oxidase/dehydrogenase activity (CKX) in the tissues of young pea plants was studied. Tested anticytokinins, with the exception of the most efficient one – PU-1, were able to promote increased CKX activity in roots, when applied alone, but they had no significant influence on the enzymatic activity in leaves. N⁶-benzyladenine (BA) and 1-(2-chloropyridin-4-yl)-3-phenylurea (CPPU) provoked strong increase in CKX activity in roots, while in leaves considerable inhibition of enzymatic activity was observed. Different types of anticytokinins exhibited diverse preference towards taking off the action of purine and phenylurea cytokinins over CKX activity.