石榴果色合成相关基因CHS和CHI的表达特性分析

花色苷是类黄酮家族中重要的一类次生代谢产物,对果实呈色起重要作用。CHS (查尔酮合成酶)和CHI (查尔酮异构酶)为花色苷合成提供了前体物质,是花色苷合成所不可或缺的。利用RT-PCR和RACE方法,本研究从石榴果皮中克隆了与花色苷合成相关的CHS基因和CHI基因的cDNA全长,同时采用qRT-PCR研究了这两个基因在三个不同色泽石榴品种‘红宝石’、‘水晶甜’、‘墨石榴’发育期内的表达模式,并分析了果皮花色苷含量变化与基因转录水平的关系。结果表明,石榴中CHS和CHI基因cDNA全长分别为1 197 bp和693 bp,分别编码398和230个氨基酸,命名为PgCHS和PgCHI,在GenBank中的登录号分别为KF841615和KF841616。在氨基酸水平上,Pg CHS与荔枝、葡萄、山竹等果树的同源性达到90%以上。Pg CHI与果树中龙眼、梨、美洲葡萄、桑树等同源性达到70%以上。qRT-PCR结果显示,CHS和CHI基因的表达模式随色泽发育期和品种不同而有差异。在‘红宝石’石榴中,该两个基因都有前期和后期两个表达高峰期;而‘水晶甜’石榴中这两个基因的表达高峰期均出现在中后期;‘墨石榴’发育初期时CHS和CHI的表达量最高,以后的表达量都较低。同一品种内,CHS和CHI的表达具有协同性,两者的协同性表达有利于花色苷及其他类黄酮相关产物的合成。3个品种中CHS和CHI基因的表达与花色苷的积累并不一致。     

‘脆红李’果实发育过程中花色苷含量及相关基因表达分析

花色苷是一类重要的色素,对李红色的形成必不可少。本研究以‘脆红李’为试材,研究了果实发育过程中叶绿素含量、总花色苷含量及果皮主要花色苷组分和含量的变化规律,并分析了Ps PAL、Ps CHS、Ps CHI、Ps F3H、Ps DFR、Ps ANS和Ps UFGT基因在果实不同发育阶段的表达规律。结果表明,随着‘脆红李’果实的生长发育,果皮和果肉中总叶绿素含量呈逐渐下降的趋势;‘脆红李’果肉中不含花色苷,果皮中的花色苷在转色期才开始积累,成熟时达到最大值,为404.37μg/(g·FW),并以矢车菊素-3-O-葡萄糖苷和矢车菊素-3-O-芸香糖苷为主;花色苷合成相关基因在‘脆红李’不同生长发育时期的果皮和果肉中有着特异性的表达,但只有Ps PAL和Ps UFGT基因的转录水平与花色苷含量的正相关性达到极显著水平,表明这两个基因对‘脆红李’果实的着色有着异常重要的调控作用。     

不同红梨果皮类黄酮合成基因表达模式分析

采用半定量和荧光定量PCR方法,分析10个类黄酮合成基因在梨品种‘红星’和‘满天红’成熟果皮中的转录特性以及光照对基因表达的影响.结果表明:‘满天红’花色苷合成上游基因(CHS、CHI)的表达量高于‘红星',而下游基因(F3H、DFR、ANS)以及黄酮醇(FLS)和原花色素(LAR、ANR)合成相关基因的表达量却正好相反.套袋去除光照可使所有被检测基因的转录水平降低,F3GT和FLS最明显,表达量差异达20～30倍以上,且套袋‘红星’中PAL、F3H、DFR、ANS、LAR、ANR基因的表达量仍高于不套袋‘满天红’.研究认为,花色苷合成下游基因转录水平的差异是2个红色梨品种间着色不同的主要原因,而F3GT是光照调控‘红星’着色的关键基因.     

不同花色菊花品种花色素结构基因的表达特性

对红色、黄色、粉紫色和白色菊花品种不同开放度的花序舌状花中CHS、CHI、DFR、F3H、F3′H和3GT基因的表达量进行了相对定量分析。结果表显示:6个基因的表达因不同花色、不同发育阶段而异。‘钟山红鹰’(红色)中各基因的表达量均较高,且均在Ⅱ(松蕾期)或Ⅲ(半开期)期达到峰值,其中DFR、3GT基因的表达量远高于其他花色品种。‘金陵娇黄’(黄色)中CHS、CHI基因表达量较高,且Ⅰ(紧蕾期)、Ⅱ期表达量高于Ⅲ、Ⅳ(盛开期)期;3GT、DFR基因表达量分别高或低于‘金陵笑靥’(粉紫色)品种中相应基因的表达量,但均比红色品种低;F3H在4个品种中表达量最低,F3′H表达量接近或略低于红色或粉紫色品种,且各阶段表达水平较稳定。‘金陵笑靥’中DFR表达量仅次于‘钟山红鹰’,3GT和CHS表达量低于红色与黄色品种。‘钟山雪桂’(白色)中各基因仅有微量表达,除F3H外各基因的表达量明显低于其他花色品种。研究表明,花色素结构基因DFR、3GT是菊花花色素合成的关键基因,DFR很可能是限速关键基因,一定表达水平的CHS、CHI也是菊花花色素合成所必须的,F3H基因与花色素合成不存在直接相关。     

葡萄果实成熟期花色苷与白藜芦醇合成相关基因的表达

以酿酒葡萄‘赤霞珠’为试材,采用pH示差法和高效液相色谱(HPLC)法分别测定葡萄成熟期果皮花色苷和白藜芦醇含量,用实时荧光定量PCR检测两者合成途径中相关基因的表达量,分析花色苷含量和白藜芦醇含量与其相关基因表达的关系,以揭示结构基因与调控基因的调控机制,为筛选富含花色苷和白藜芦醇的酿酒葡萄提供理论依据。结果显示:(1)葡萄果皮花色苷含量在花后112d达到最高值(0.77mg/g),反式白藜芦醇含量在花后126d达到最高值(30.87μg/g)。(2)花色苷和白藜芦醇合成途径中,CHSs、CHI、STS、UFGT、MybA1、MybA2基因的表达量除花后98d下调外,其余时间均呈上调表达,而Myb5a则始终呈上调表达。(3)相关分析表明,STS基因表达量与CHS1、CHS2基因表达量呈极显著和显著正相关关系,MybA1、MybA2基因表达量与CHSs、CHI、STS、UFGT基因的表达量呈极显著正相关关系;Myb5a基因表达量与CHS3基因表达量呈极显著正相关关系。研究表明,部分结构基因的表达与花色苷和白藜芦醇的变化不同步,MybA1和MybA2可能调控花色苷合成途径中多个结构基因的表达,花色苷与白藜芦醇的关系并不固定,而是处在动态变化中。     

‘津田芜菁’花色素苷生物合成相关基因的表达

利用黑暗、日光、人工恒定光处理花色素苷合成依光型的‘津田芜菁’试材。通过紫外．可见分光光度计测定恒定光处理下‘津田芜菁’块根皮中花色素苷的含量,结果表明,花色素苷含量与光处理时间成正相关。用从消减文库中筛选的花色素苷生物合成基因片段制备探针,Northern杂交结果显示,在所处理的48h之内,光可以诱导‘津田芜菁’中PAL、DFR、CHS、F3H和ANS基因的表达,这些基因的表达量随着光处理时间的延长而增加,而MYB基因的表达量在黑暗与光下基本相同。     

花青素生物合成关键酶的研究进展

花青素是植物花呈现不同色彩的物质基础,其生物合成途径主要受到查尔酮合成酶(CHS)、查尔酮异构酶(CHI)、黄烷酮3-羟化酶(F3H)、类黄酮3'-羟化酶(F3'H)、类黄酮3’,5’-羟化酶(F3'5'H)、二羟基黄酮醇还原酶(DFR)、花色素苷合成酶(ANS)以及类黄酮3-O-糖基转移酶(UFGT)等关键酶的控制.主要介绍花青苷生物合成途径、关键酶晶体结构及利用基因工程改造花色的研究进展,讨论目前花色改造存在的问题,并对今后的研究前景进行展望.     

葡萄‘霞多丽’果实中黄烷-3-醇合成酶活性及其相关基因表达北大核心CSCD

为探究酿酒葡萄果实中黄烷-3-醇积累规律与合成相关酶、关键基因表达的关系,该研究以贺兰山东麓主栽酿酒白葡萄品种‘霞多丽’为材料,采用比色法、高效液相色谱(HPLC)和荧光定量PCR技术,分析葡萄果实发育过程中黄烷-3-醇单体物质的积累规律及其生物合成过程中相关酶活性、相关基因表达模式的变化。结果表明:(1)在‘霞多丽’果实整个发育期,黄烷-3-醇主要单体物质没食子儿茶素(GC)、表儿茶素没食子酸酯(ECG)含量远高于儿茶素(CAT)、表儿茶素(EC)含量,且ECG、EC含量均随果实发育期进程呈先升后降变化趋势,并于花后50 d(7月13日)达最大值,分别为47.69和42.73 mg/g,而GC、CAT的含量则呈先降低后升高,后随果实成熟逐渐降低的变化趋势。(2)葡萄果实中黄烷-3-醇生物合成相关酶PAL和ANS活性随果实发育期进程呈先升高后降低的变化趋势,F3H、F3′H、CHI活性均在花后20 d(6月13)出现峰值,而后活性迅速降低,而DFR活性呈先升高后降低,并于花后50 d(7月13)再次升高,后随果实成熟逐渐降低。(3)黄烷-3-醇生物合成相关结构基因PAL、ANS的表达水平随果实发育期进程呈先上调后下调的趋势,与ECG、EC含量变化趋势一致,F3′H、F3H、CHI的表达水平却呈先下调后上调的趋势,DFR的表达水平先上调而后下调,并随果实成熟再次上调。(4)相关性分析表明,F3H、F3′H和ANS酶活性与GC含量呈显著正相关关系;F3H、F3′H基因表达与GC、CAT的含量呈正相关关系,ANS、PAL基因表达与ECG、GC的含量呈正相关关系。研究发现,黄烷-3-醇生物合成相关酶和相关基因表达影响了葡萄果实中黄烷-3-醇类化合物含量的积累,从而进一步影响葡萄酒的感官品质。     

石榴PgUFGT基因克隆及表达分析

利用RT-PCR和RACE方法,从石榴(Punica granatum L.)果皮中克隆到一个类黄酮糖基转移酶(UFGT)基因(PgUFGT)全长cDNA序列(GenBank登录号为KF841620)。PgUFGT基因编码区1 476bp,编码491个氨基酸。PgUFGT蛋白具有保守PSPG基序、UDP-糖基转移酶家族结构域和UDP-葡萄糖醛酸基/葡萄糖基转移酶保守域(UDPGT),与其他植物UFGT蛋白一致性较高;系统进化树分析结果表明,PgUFGT属于类黄酮3-O-糖基转移酶类。荧光定量qRT-PCR结果表明,PgUFGT基因在‘红宝石’和‘水晶甜’2个石榴品种的发育期内具有不同的表达模式,PgUFGT在‘红宝石’石榴中有2个转录表达高峰,而在‘水晶甜’石榴中仅有1个表达高峰,表明PgUFGT可能在2个石榴品种中具有不同的催化作用。该研究结果为进一步研究石榴果实色泽形成的分子机制奠定了基础。     

不同栽培区域和年份的红皮砂梨果皮着色特性

该研究以红皮砂梨品种‘八月红’和‘红香酥’为试材,比较了不同栽培地区和年份的红皮砂梨着色规律、花色素苷组分及含量,以及相关代谢酶的变化。结果显示:（1）‘八月红’和‘红香酥’果皮花色素苷含量变化规律由其品种特性决定,同一品种的果皮花色素苷绝对含量受不同栽培区域和年份的影响,其组成和含量变化趋势不受影响;但不同品种的花色素苷含量变化以及着色规律有所不同。（2）红色砂梨果皮着色主要与矢车菊-3-半乳糖苷含量变化一致,且随着梨果实的发育和着色加深,果实中UDP-葡萄糖:类黄酮-3-O葡萄糖苷转移酶（UFGT）、二氢黄酮醇-4-还原酶（DFR）活性呈上升趋势,而苯丙氨酸解氨酶(PAL)活性随着果实发育而下降。（3）相关性分析表明,两品种梨果实花色苷含量与其UFGT、DFR活性呈显著或极显著正相关关系,而与PAL活性相关性不一致。研究表明,梨果皮中的UFGT、DFR是影响红皮砂梨着色的重要酶。     

六种藓类植物茎的比较解剖学研究

通过对6种藓类植物,即褶叶青藓(Brachythecium salebrosum(Web.et Mohr.)B.S.G.)、湿地匐灯藓(Plagiomnium acutum(Lindb.)Kop.)、侧枝匐灯藓(Plagiomnium maximoviczii(Lindb.)Kop.)、大凤尾藓(Fissidensnobilis Griff.)、大羽藓(Thuidium cymbifolium(Doz.et Molk.)B.S.G.)和大灰藓(Hypnum plumaeforme Wils.)嫩茎和老茎的石蜡切片和显微观察发现,同一藓类植株的嫩茎和老茎,茎结构稳定,不同种藓类植物茎横切面具有不同特征.植物体茎横切面形状、表层细胞的层数、细胞大小和细胞壁厚薄、皮层细胞大小和形状、中轴的有无以及比例等特征可以作为藓类植物的分科分类依据之一.     

Cause of Senescence of Nine Sorts of Flowers

The levels of endogenous phytohormones and respiratory rate in nine sorts of flowers such as Cymbidium faberi Rolfe, Nopalxochia ackermannii Kunth and others were investigated both at full bloom and senescence and meanwhile the effect of exogenous phytohormones on prolonging the blossoms and promoting ethylene production were tested. There is a high content of endogenous ethylene in all the long-lived flowere, about 3–16 folds higer than the short-lived ones. There is a high level of ABA at full blooming flowers of short-lived flowers, in which there is no or only some cytokinins in it, but the ratio of CTK (6BA+zeatin)/ABA is smaller(l.7). The endogenous ABA reached a much higher level at senescence in all nine sorts of flowers, so it is reasonable to consider that it is ABA which plays an important role of regulation in controlling flower's senescence. There is a much higher level of GA3 and zeatin in the long-lived flowers which is not demonstrated in the shortlived ones. The respiratory rate is one of the factors controtling the longevity of flowers, but it does not play a decided role. Application of 6BA and zeatin prolongs distinctly orchid’s longevity, however exogenous IAA through the promotive action on ethylene production, evidently extends the longevity of the flowers of the Nopalxochia ackermannii Kunth.     

Photoregulation of seed germination of wild-type and of an aurea-mutant of tomato

Seed germination of an aurea mutant of tomato ( Lycopersicon esculentum Mill.) is promoted by continuous irradiation with red, far-red or long-wavelength far-red (758 nm) light as well as by cyclic irradiations (5 min red or 5 min far-red/25 min darkness). Far-red light applied immediately after each red does not change the germination behaviour. Seed germination of the isogenic wild-type, cv. UC-105, is promoted by continuous and cyclic red light while it is inhibited by continuous and cyclic far-red light and by continious 758 nm irradiation. Far-red irradiation reverses almost completely the promoting effect of red light. The promoting effect (in the aurea mutant) and the inhibitory effect (in the wild-type) of continuous far-red light do not show photon fluence rate dependency above 20 nmol m⁻² s⁻¹. It is concluded that phytochrome controls tomato seed germination throgh low energy responses in both the wild type and the au mutant. The promoting effect of continuous and cyclic far-red light in the au mutant can be attributed to a greater sensitivity to P_fr.     

龙胆科药用植物化学成分的研究现状

龙胆科植物在我国的分布范围很广,且多数为药用植物,其多数种属的药用植物,至今其化学成分尚未被系统研究。综述了目前龙胆科药用植物的化学成分的研究现状及一般提取方法,对近年来发现的环烯醚萜及裂环烯醚萜类化合物进行了总结,为本科药用植物的更深入研究提供了参考。     

The effects of glutamine of the maintenance of embryogenic cultures of Cryptomeria japonica

Summary Embryogenic tissues of sugi (Cryptomeria japonica) were induced on a modified Campbell and Durzan (CD) medium containing 1 μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 600 mg l⁻¹ glutamine, and subcultured in the medium of the same composition for over 1 yr. This resulted in a mixed culture of embryogenic and non-embryogenic cells. When embryogenic cells were isolated and cultured independently, their capacity to form embryogenic aggregates was lost. Thus, the non-embryogenic cells present within a mixed culture system were essential to the formation of embryogenic aggregates. When embryogenic tissues were isolated and cultured independently on a high glutamine-containing (2400 mg l⁻¹) medium, dry weights and endogenous levels of glutamine increased, and the tissue could generate a large number of embryogenic aggregates. Amino acid analysis of embryogenic and non-embryogenic cells from the maintenance culture indicated a higher level of glutamine was present in the latter. The high endogenous level of glutamine in the non-embryogenic portion of mixed cell masses may be the supplier of glutamine for maintaining the embryogenic property of the tissues.     

Specificity of action of piperidylcholine derivatives as substrates of cholinesterases of various origin

The review deals with study of enzymologic properties of a novel highly specific acetylcholinesterase substrate, N-(β-acetoxyethyl) piperidinium iodomethylate (“piperidylcholine”), and its 30 derivatives that were tested as effectors of cholinesterases of mammals and various species of Pacific squids. It was proven for the first time that responsible for specificity of action was structure of cyclic ammonium grouping of the alcohol part of molecule of the ester substrate. Analysis of specificity is performed based on enzymatic hydrolysis parameters—activity of catalytic center of cholinesterases and bimolecular constant of the reaction rate that are determined at optimal and low substrate concentrations. Among the specially synthesized group of thioester compounds there is revealed one more highly specific acetylcholinesterase substrate—N-(β-acetoxyethyl) piperidinium.     

Interconnection of parameters of regulation system of lipid peroxidation and of morphophysiological parameters of mouse liver

A complex analysis of seasonal fluctuations of the mean group parameters of the system of regulation of lipid peroxidation has been performed in liver of Balb/c mice. Association of lipid characteristics and morphophysiological parameters is studied in the Balb/c mouse liver. An inter-connection is revealed between the liver index and the amount of lysoforms of phospholipids, the scale and character of the interconnection differing essentially depending on proportion of phos-phatidylcholine in mouse liver phospholipids.     

真菌类遗传学分析的知识结构教学

本文以认知结构理论为指导,讨论了真菌类遗传分析与高等动植物遗传分析的内在联系,认为利用这种内在联系进行教学可收到好的效果并说明了作者的具体教学过程。 Abstract:In the paper, the relationship between genetic analysis of Fungi and genetic analysis of high animal and plant was discussed.A good results were obtained when we adopted this method in the teaching.