高等植物脱落酸生物合成途径及其酶调控

脱落酸(ABA)生物合成一般有两条途径:C15直接途径和C40间接途径,前者经C15法呢焦磷酸(FPP)直接形成ABA;后者经由类胡萝卜素的氧化裂解间接形成ABA,是高等植物ABA生物合成的主要途径.9-顺式环氧类胡萝卜素氧化裂解为黄质醛是植物ABA生物合成的关键步骤,然后黄质醛被氧化形成一种酮,该过程需NAD为辅因子,酮再转变形成ABA-醛,ABA-醛氧化最终形成ABA.在该途径中,玉米黄质环氧化酶(ZEP)、9-顺式环氧类胡萝卜素双加氧酶(NCED)和醛氧化酶(AO)可能起重要作用.     

高等植物脱落酸生物合成途径及其酶调控

脱落酸（ABA）生物合成一般有两条途径：C15直接途径和C40间接途径, 前者经C15法呢焦磷酸（FPP）直接形成ABA;后者经由类胡萝卜素的氧化裂解间接形成ABA, 是高等植物ABA生物合成的主要途径。9-顺式环氧类胡萝卜素氧化裂解为黄质醛是植物ABA生物合成的关键步骤, 然后黄质醛被氧化形成一种酮, 该过程需NAD为辅因子, 酮再转变形成ABA-醛, ABA-醛氧化最终形成ABA。在该途径中,玉米黄质环氧化酶（ZEP）、9-顺式环氧类胡萝卜素双加氧酶（NCED）和醛氧化酶（AO）可能起重要作用。     

大蜡螟成虫3个醛氧化酶基因的鉴定、序列特征与组织表达模式

醛氧化酶(AOXs)在昆虫的嗅觉生理代谢过程中起重要作用.本研究从大蜡螟Galleria mellonella成虫中鉴定了3个AOXs基因,命名为GmelAOX1、GmelAOX2和GmelAOX3.这3个基因均含有完整的开放阅读框,所编码的蛋白质均具有醛氧化酶的典型特征,如具有铁硫氧化还原中心、黄素腺嘌呤二核苷酸结合区域和钼辅因子结合区域.系统进化分析显示3个GmelAOXs被聚在不同的进化分支,且GmelAOXs与鳞翅目AOXs亲缘关系最近.GmelAOX1和GmelAOX3位于同一个基因组框架(scaffold69)上,而GmelAOX2位于scaffold96.GmelAOX1、GmelAOX2和GmelAOX3的外显子数量分别为17个、16个和21个.GmelAOX2高量表达于成虫触角,且在触角中的表达量显著高于其它组织,推测GmelAOX2可能编码一个气味降解酶并参与醛类气味分子的降解.GmelAOX1和GmelAOX3的表达没有组织特异性,二者编码的蛋白可能兼具气味降解和外源醛类代谢的功能.     

芒果生长素反应因子类蛋白的cDNA克隆和表达

通过SSH法获得了一个与不定根形成相关的差异表达的cDNA片段,其推导的氨基酸序列与拟南芥的生长素反应因子(ARF)类蛋白具有较大的同源性,因此将它命名为MiARF。用所设计的基因特异引物进行3′RACE扩增获得包含完整读码框架(ORF)的MiARF1（GenBank登录号为AY255705）和MiARF2（GenBank登录号为AY300808）。MiARF1全长为3272bp,其中,ORF含2523bp,5′非翻译区（5′UTR）含285bp, 3′非翻译区(3′UTR)含464bp。由该序列所推导的氨基酸序列与拟南芥ARF2（BAB10162）的ID值为64%, E值为0,在DNA结合区域（DBD）、III和IV区域的同源性更高,ID值均大于80%。MiARF2cDNA全长为1474bp,其中ORF含981bp,5′非翻译区含285bp, 3′非翻译区含208bp,由该序列所推导的氨基酸序列与拟南芥ARF2（BAB10162）的ID值为84%,E值为e-151。 MiARF2仅具有DBD保守区并与MiARF1的基本相同,但缺乏III和IV区域。Virtural Northern 杂交表明：MiARF2在生根的组织中表达水平高, 而在非生根的组织中未见表达;MiARF1在生根及非生根的组织中均有表达。     

RACE法克隆甜菜NR基因及生物信息学分析

硝酸还原酶是氮素代谢的关键酶和限速酶,研究硝酸还原酶的功能对提高甜菜的产量具有重要作用。运用RACE法克隆出甜菜的硝酸还原酶基因,并利用生物信息学对甜菜NR进行主要结构分析和功能预测,并使其在拟南芥中表达,观察根对向重性应答过程中的弯曲情况。结果表〖JP2〗明,利用RACE法克隆得到甜菜NR cDNA全长为2 760 bp;甜菜NR为易溶、亲水性强的蛋白;二级结构预测结果显示,甜菜NR为混合型蛋白;甜菜NR含有钼辅因子、细胞色素b5、FAD及NADH结合域,具有跨膜区域,但不含有信号肽;利用拟南芥突变体观察到野生型比突变体的弯曲较快,暗示甜菜的NR基因可能通过调节NO积〖JP〗累参与植物向重性应答。     

西番莲PeERG基因克隆及其表达模式分析

ERA(Eecherichia coli Ras-like protein)蛋白是与已知异三聚体G蛋白和小分子G蛋白不同的一种新的GTP结合蛋白。为了在木本植物中开展其同源基因ERG(ERA-like GTPase)克隆和功能验证的相关研究,该文首次在西番莲新品种‘平塘1号’中采用cDNA末端快速克隆(RACE)技术克隆鉴定1个ERG基因。结果表明:西番莲PeERG基因cDNA全长为1 518 bp,包括1 260 bp的开放阅读框、38 bp的5'-端非翻译区和220 bp的3'-端非翻译区,该基因编码蛋白由420个氨基酸残基组成,其二级结构含有丰富的α-螺旋和延伸链。PeERG蛋白不含跨膜区域,也不存在信号肽酶切位点,既在其N端有典型的GTPase保守结构域(GTPase domain)又在其C端有独特的RNA结合结构域(KH domain)。系统进化树分析表明,西番莲PeERG蛋白和水稻OsERG1、拟南芥AtERG1、大肠杆菌ERA位于同一进化分枝。实时定量PCR检测揭示PeERG基因在西番莲根、茎、叶、花、果中均有表达,叶中表达最高;同时该基因响应低温胁迫信号,其表达呈动态变化模式。该研究首次鉴定和描述了木本植物西番莲的ERG基因,为深入挖掘西番莲特异基因资源提供参考,也有助于进一步探究ERG基因在植物中的生物学功能及其作用机制。     

家蚕醛氧化酶基因（BmAOXs）的鉴定与表达分析

醛氧化酶（aldehyde oxidases, AO, EC 1.2.3.1）是属于钼-黄素酶（molybdo-flavoenzymes, MFEs）家族的一类蛋白酶。为了探讨该酶在家蚕Bombyx mori中的功能, 本研究对家蚕醛氧化酶基因（BmAOXs）家族进行了鉴定和分析。以其他物种AO基因序列检索家蚕全基因组数据库, 获得了8个BmAOX候选基因, 均具有醛氧化酶保守的功能域。进化分析表明, BmAOX与其他昆虫AO聚为一簇。RT-PCR分析结果显示: BmAOX1, BmAOX2, BmAOX3, BmAOX5具有很强的组织特异性; 而BmAOX4, BmAOX6, BmAOX7, BmAOX8则在蛹和成虫的多个组织中均有表达, 提示它们在家蚕生理代谢活动中起重要作用。利用Native PAGE和活性染色方法, 对BmAOX编码的蛋白进行检测, 结果表明: 家蚕蛹中有5种有活性的醛氧化酶, 而成虫中有6种, 各组织中均有有活性的BmAOX, 只是种类和活性水平有所不同。本研究结果为深入探讨BmAOX家族的功能奠定了基础。     

高等植物脱落酸生物合成的酶调控

高等植物ABA的生物合成开始于细胞质内的甲瓦龙酸 (MVA)或位于叶绿体内的丙酮酸_硫胺素焦磷酸 (TPP) ,经一系列反应最后在质体或胞质中形成的。除胁迫或植物发育中生理变化引起的诱导外 ,ABA的合成还受到一系列酶的调控 ,其中 ,玉米黄质环氧化酶 (ZE) ,9_顺环氧类胡萝卜素双加氧酶(NCED)和醛氧化酶 (AO)可能起到重要的调节作用。本文介绍近年来ABA生物合成酶调控的研究进展。     

高等植物脱落酸生物合成的酶调控

高等植物ABA 的生物合成开始于细胞质内的甲瓦龙酸（MVA）或位于叶绿体内的丙酮酸_硫胺素焦磷酸（TPP）,经一系列反应最后在质体或胞质中形成的。除胁迫或植物发育中生理变化引起的诱导外,ABA的合成还受到一系列酶的调控,其中,玉米黄质环氧化酶（ZE）,9_顺环氧类胡萝卜素双加氧酶（NCED）和醛氧化酶（AO）可能起到重要的调节作用。本文介绍近年来ABA生物合成酶调控的研究进展。     

拟南芥干旱诱导型启动子RD29A驱动花生AhNCED1基因双元表达载体的构建

从拟南芥基因组中克隆RD29A基因5'-侧翼520bp启动子区域序列,生物信息学分析表明,该启动子片段中存在脱水胁迫响应元件(DRE)、ABA响应元件(ABRE)、TATA-box、CAAT-box等顺式作用元件。构建了干旱诱导型启动子AtRD29Ap驱动花生AhNCED1基因的植物双元表达载体pAtRD29Ap::AhNCED1。     

葡萄果实中脱落酸结合蛋白的存在及其性质

葡萄果实微粒体上存在高亲和力的脱落酸（ＡＢＡ）结合位点,这些位点与ＡＢＡ的结合具有饱和性,高亲和力及低容量,胰蛋白酶或ＤＴＴ处理可以使该位点的特异结合活性下降约９０％,表明此结合位点是一种蛋白质,故称为ＡＢＡ结合蛋白,它含有维系蛋白质特定构象的二硫键,该蛋白与ＡＢＡ反应的最适ｐＨ为６．０,说明与配基结合部位可能存在带有正电荷的氨基酸残基,结合活性在２５℃高于０℃,结合反应达到动态平衡需要３０ｍｉｎ,３０ｍｉｎ以后结合活性随时间延长而下降。该蛋白与ＡＢＡ结合反应的平衡解离常数为１７．５ｎｍｏｌ／Ｌ,最大结合容量（Ｂｍａｘ）为９８．４ｆｍｏｌ／ｍｇｐｒｏｔｅｉｎ。     

大豆花荚败育期间的植物激素变化

大豆开花结荚期,不同发育阶段的幼蕾与花荚的脱落率不同,其中以花后５ｄ内的幼荚脱落最严重。与败育花荚相比,正常花荚中的干物质积累量均较高。细胞分裂素（ＤＨＺＲｓ,ＺＲｓ,ｉＰＡ）含量也较高,花后３～５ｄ的幼荚中表现更明显。脱落酸（ＡＢＡ）则是以败育幼蕾及花后３～５ｄ的幼荚中含量较高。不同发育阶段的大豆生殖器官中,正常开放花中的玉米赤霉烯酮（ＺＥＮ）含量最高     

Synthesis and biological activity of abscisic acid esters

16 ABA esters including 11 new compounds were prepared by two different esterification routes. All the structures of ABA esters were confirmed by ¹H NMR, ¹³C NMR and HRMS. Their biological activity and hydrolysis stability were investigated. Fortunately, there were 15 and 9 compounds which displayed much better or nearly the same inhibition activity for rice seedling growth and Arabidopsis thaliana seed germination compared to ABA, respectively. Especially, compounds 2d and 2g showed better biological activities than ABA in the three tests. Moreover, we found that chemical hydrolysis ability of the esters in vitro had little relationship to their biological activity.     

ABA 对黑黄檀种子萌发的抑制作用以及其他植物激素对ABA 的拮抗作用

以云南特有濒危树种黑黄檀( Dalbergia fusca) 的种子为材料, 研究了脱落酸(ABA) 对种子萌发的抑制作用, 以及种子萌发过程中吲哚乙酸( IAA) 、赤霉酸(GA3 )、6-苄基腺嘌呤(6-BA) 和乙烯利对ABA的拮抗作用。黑黄檀种子萌发的适宜温度为30℃。交替光照(14 h 光照和10 h 黑暗) 以及黑暗对种子萌发没有明显的影响。0 . 001～0 . 1 mmol/L ABA 不影响种子的萌发率, 但降低种子的萌发进程; 1 mmol􊄯L 和2 . 5mmo􊄯l L ABA 显著地抑制种子的萌发率和萌发进程。种子的萌发率不被0 . 0001 ～ 1 mmo􊄯l L IAA 和GA3 、0 . 0001～0 . 1 mmol/L 6-BA、以及0 . 001～10 mmol/L 乙烯利( 乙烯供体) 的影响,但被1 mmol􊄯L 6-BA 抑制。1mmol/L ABA 对种子萌发的抑制作用能被0 . 01～1 mmol/L IAA、0 . 01～1 mmol/L GA3 、0 . 001～0 .1 mmol/L 6-BA 和0 . 1～10 mmol􊄯L 乙烯利所拮抗, 而且这种拮抗作用与植物激素的类型和浓度有关。0. 01 mmol/L 6-BA 和0 . 1 mmol/L 乙烯利对1mmol/L ABA 抑制作用的拮抗不能被添加0 . 001 mmol/L IAA 或者0 .001 mmol/L GA3 加成。但0 . 1mmol/L 乙烯利对1 mmol/L ABA 抑制作用的拮抗能够被添加0 . 01 mmol/L 6-BA 或者0 .1 mmol/L 6-BA 加成, 导致更高的萌发率和幼苗生长。     

脱落酸受体及相关信号传导的研究进展

对脱落酸信号传导的研究主要集中在种子成熟和休眠和气孔的运动上。研究者对脱落酸受体上作了大量的工作,但很长时间来仍没有发现脱落酸受体基因。最近,脱落酸受体的研究有了重大突破。研究者在拟南芥中发现FCA和CHLH两个脱落酸受体基因。     

Radioimmunoassays for the differential and direct analysis of free and conjugated abscisic acid in plant extracts

Two radioimmunoassays have been developed which allow the parallel quantitation of free as well as conjugated natural (+)-abscisic acid (ABA) directly and separately, in unpurified plant extracts. The differential specificity of antisera has been achieved by coupling ABA through C₁ (for total ABA determination) or C₄ (for free ABA determination), respectively, to proteins to obtain the immunogenic conjugates. Compounds structurally related to ABA, such as, dihydrophaseic acid or phaseic acid, do not interfere with either of the assays, even when present in more than ten-fold excess. Other related compounds, such as, violaxanthin or xanthoxin, do not cross react at all. Both antisera respond to (+)-ABA but show very low immunoreactivity with (-)-ABA. As little as 27 pg of ABA (serum for free ABA) or 47 pg (serum for total ABA) may be detected and the measuring ranges are from 0.2–8 and 0.2–30 pmol, respectively. Average recoveries are greater than 99%. Using these assays, more than 100 samples can be assayed for free and conjugated ABA per day. Levels of free ABA, as determined by radioimmunoassay (RIA), correlated well with those reported in the literature. Levels of conjugated ABA were found to be generally higher than previously reported for ABA after alkaline hydrolysis of the extracts. Conjugated ABA accumulates during aging of leaves and levels of conjugated ABA up to 17-fold higher than those of free ABA have been detected in senescent leaves of Hyoscyamus niger L. Evidence was obtained for the presence of ABA conjugates other than the glucose ester in some plants.Abbreviations ABA abscisic acid - BHT 2,6-di-t-4-methyl phenol - BSA bovine serum albumin - HSA human serum albumin - RIA radioimmunoassay - TLC thin-layer chromatography - EDC 1-ethyl-3(3-dimethylaminopropyl)-carbodiimide · HCl Part 11 in the series: Use of Immunoassay in Plant Science     

Stereoselectivity in the biosynthetic conversion of xanthoxin into abscisic acid

All stereoisomers of xanthoxin (XAN) and abscisic aldehyde (ABA-aldehyde) were prepared from (R) and (S)-4-hydroxy--cyclogeraniol via asymmetric epoxidation. Their stomatal closure activities were measured on epidermal strips of Commelina communis L. Natural (S)-ABA-aldehyde showed strong activity comparable to that of (S)-abscisic acid (ABA). Natural (1S, 2R, 4S)XAN and (1S, 2R, 4R)-epi-XAN also induced stomatal closure at high concentrations. On the other hand, unnatural (1R)-enantiomers of XAN, epi-XAN, and ABA-aldehyde were not effective. To further examine the Stereoselectivity on the biosynthetic pathway to ABA, deuterium-labeled substrates were prepared and fed to Lycopersicon esculentum Mill, under non-stressed or water-stressed conditions. Substantial incorporations into ABA were observed in the cases of natural (1S, 2R, 4S)-XAN, (1S, 2R, 4R)-epi-XAN and both enantiomers of ABA-aldehyde, leading to the following conclusions. The negligible effect of unnatural (1R)-enantiomers of XAN, epi-XAN and ABA-aldehyde can be explained by their own biological inactivity and/or their conversion to inactive (R)-ABA. Even in the isolated epidermal strips, putative aldehyde oxidase activity is apparently sufficient to convert ABA-aldehyde to ABA while the activity of XAN dehydrogenase seems very weak. The stereochemistry of the 1, 2-epoxide is very important for the XAN-dehydrogenase while this enzyme is less selective regarding the 4-hydrdxyl group of XAN and converts both (1S, 2R, 4S)-XAN and (1S, 2R, 4R)-epi-XAN to (S)-ABA-aldehyde. Abscisic aldehyde oxidase can nonstereoselectively convert both (S) and (R)-ABA-aldehyde to biologically active (S) and inactive (R)-ABA, respectively.Abbreviations ABA abscisic acid - ABA-aldehyde abscisic aldehyde - DET diethyl tartrate - epi-XAN xanthoxin epimer - FCC flash column chromatography - GC-EI-MS gas chromatography-electron impact-mass spectrometry - MeABA abscisic acid methyl ester - IR infrared - NMR nuclear magnetic resonance - PCC pyridinium chlorochromate - THF tetrahydrofuran - XAN xanthoxin The authors are very grateful to Mr J.K. Heald (Department of Biological Sciences, University of Wales, Aberystwyth, UK) and Dr. R. Horgan for carrying out GC-EI-MS analyses and advice, respectively.This work was supported by the Japan Society for the Promotion of Science (Fellowship for Young Japanese Researcher No. 0040672).     

Problems and Possibilities in the Teaching of Ecology

The treatment of technical vocabulary largely deter-mines its effectiveness as a medium of communication. The present paper describes how six O-level1 Biology texts were analysed, to discover how they dealt with terminology. Two categories of vocabulary were recognised: the terms peculiar to individual books and those common to all books. Individual vocabu-laries were of different sizes because of divergences in style and the use of various synonyms to express the same idea. (Synonyms also increase vocabulary burden and make it more difficult for a pupil to pass from one book to another.) The common vocabulary was thought to contain the most important terms, and the authors' treatment of them seemed to bear this out: a large majority were both explained and emphasised. Technical vocabulary would be used even more effectively if synonyms were eliminated and all the remaining terms were explained and emphasised at their first appearance.     

Stomatal responses to rapidly imposed water stress and light/dark transition in norflurazon-treated wheat leaves

Hoglund, H. O. and Klockare, R. 1987. Stomatal responses to rapidly imposed water stress and light/dark transition in norflurazon-treated wheat leaves.
Stomatal responses to rapidly imposed water stress and to light/dark transition were studied in leaves of wheat ( Triticum aestivum L. cv. Starke II) treated with nor-flurazon (NF) which is known to inhibit abscisic acid (ABA) accumulation. The stomatal response was studied in an open air flow system. It was shown that these plants have the ability to respond to externally added ABA. When the water potential in the nutrient solution was rapidly reduced, stomata in green plants responded with a transient opening followed by a strongly decreased aperture. NF-treated plants responded with a similar rapid opening of stomata, but the following closure was strongly reduced. Transfer from light to darkness induced a rapid closure of stomata in green plants but the closing response was strongly delayed in NF-treated plants. These results indicate that NF affects one or more regulators involved in the closure of stomata under rapidly imposed water stress and in the light/dark transition. The possibility that this regulator is ABA is discussed.