水稻线粒体atpA基因的克隆及其与细胞质雄性不育的关系

本研究以水稻BT型细胞质雄性不育系秋光和相应的保持系秋光为材料,提取线粒体DNA,用限制性内切酶完全酶解,以玉米线粒体atpA基因和波菜叶绿体atpA基因作为探针,进行分子杂交,将保持系线粒体atpA基因定位在3.5kb的Bam HI酶切片段上,并且以pBR322为载体,克隆了这一片段,另外,在Bam HI完全酶解普带的杂交结果中,不育系线粒体基因组中有两条阳性杂交带,分别是3.5kb和2.9kb,而保持系线粒体基因组中只有3.5kb一条阳性杂交带,因而认为水稻不育系线粒体基因组中可能有两个atpA基因拷贝,而相应的保持系线粒体基因组中只有一个atpA基因拷贝。     

水稻线粒体atpA基因的克隆及其与细胞质雄性不育的关系

本研究以水稻BT型细胞质雄性不育系秋光和相应的保持系秋光为材料，提取线粒体DNA，用限制性内切酶完全酶解，以玉米线粒体atpA基因和波菜叶绿体atpA基因作为探针，进行分子杂交，将保持系线粒体atpA基因定位在3.5kb的Bam HI酶切片段上，并且以pBR322为载体，克隆了这一片段，另外，在Bam HI完全酶解普带的杂交结果中，不育系线粒体基因组中有两条阳性杂交带，分别是3.5kb和2.9kb，而保持系线粒体基因组中只有3.5kb一条阳性杂交带，因而认为水稻不育系线粒体基因组中可能有两个atpA基因拷贝，而相应的保持系线粒体基因组中只有一个atpA基因拷贝。     

应用长PCR扩增蝗虫线粒体全基因组

介绍了用两对长PCR引物扩增蝗虫(Acridoidea)线粒体全基因组的方法。从NCBI的核酸数据库下载得到36种已测昆虫线粒体全基因组,选取cytochromeb(Cytb)c、ytochrome oxidase subunitⅡ(COⅡ)、cytochrome oxidase subunitⅠ(COⅠ)基因的保守区域设计两对引物。其中引物LP03和LP04从COⅠ向Cytb扩增;引物LPCytb和LPCOⅡ从Cytb向COⅡ扩增,两对引物扩增的片段之间有大约1 kb的重叠。应用这两对引物成功扩增出10种蝗虫的线粒体基因组。考虑到在设计引物过程中所选序列在其他昆虫中的保守性,它们应能在大部分昆虫线粒体基因组扩增中发挥作用。     

氧化葡萄糖酸杆菌SCB329基因组的大小与结构的研究

收集维生素C产生菌氧化葡萄糖酸杆菌GluconobacteroxydansSCB32 9的纯培养对数期的菌体 ,采用凝胶包埋法制备完整染色体 ,用稀有酶切位点的限制性内切酶和脉冲场电泳技术对SCB32 9的基因组进行了分析 ,SpeⅠ (5′ ACTAGT)酶切有 2 4个片段 ,其大小从 1 0kb到32 0kb,用XbaⅠ (5′ TCTAGA)酶切产生 40个片段 ,其大小从 4kb到 2 0 0kb,综合两种限制酶酶切片段长度的总和结果 ,SCB32 9基因组大小为 2 70 0kb,SCB32 9基因组由一条 2 50 0kb的染色体和一个 2 4 5kb的质粒组成。通过用脱氧核糖核酸酶Ⅰ和S1核酸酶处理其基因组后电泳证实SCB32 9的染色体和质粒的拓扑学结构均为环状     

棉铃虫核型多角体病毒p40基因序列分析

克隆了棉铃虫(Helicoverpaarmigera)单粒包埋型核型多角体病毒(HaSNPV)基因组HindⅢ-H、J片段,其长度分别为10kb和6.7kb.通过对克隆片段末端测序,得到HaSNPVp40基因全序列.p40基因编码区全长966bp,预计可编码36.kD的多肽.HaSNPVp40基因核苷酸序列与HzSNPVp40基因有98%的相同性.这两种基因与BmNPVp40(GenBank-L33180)和AcMNPVgp41(GenBank-L22858)的氨基酸序列相似性43%左右,但四种蛋白对应于HzSNPVp40、HaSNPVp40的28～277氨基酸区域,同源性高达62%,并且存在一个保守的亲水性结构域.进一步将在基因组中相邻的HindⅢ-H、J两片段用BamHⅠ、EcoRⅠ、HindⅢ、PstⅠ四种限制性内切酶进行了分析.     

水稻白叶枯病菌GX1329基因组文库的构建及含编码TAL效应物基因的克隆的分离

由革兰氏阴性细菌水稻白叶枯病菌引起的水稻白叶枯病是亚洲、北美以及非洲部分地区最严重的水稻病害之一,水稻白叶枯病可使水稻减产高达50%以上.研究表明水稻白叶枯病菌的毒力主要依靠三型分泌系统所分泌的效应物.为了解水稻白叶枯病菌广西菌株GX1329中含有avrBs3/pthA家族基因的情况,本研究应用AluⅠ部分酶切其基因组DNA,构建了含有736个克隆的菌株GX1329的基因组文库.BamHⅠ酶切分析随机挑取的15个文库克隆表明,克隆的外源DNA随机性良好,克隆的最小片段为27.7 kb,最大为58.5 kb,平均大小为39.9 kb,文库克隆容量约为2.8×103 Mb,该文库中包含基因组中任一个基因的概率为99.4%.利用来自水稻白叶枯病菌菲律宾菌株PXO86的无毒基因avrXa10的第252位~第486位核苷酸序列作为探针,通过菌落原位杂交从GX1329基因组文库中筛选到37个含avrBs3/pthA家族基因的克隆.再通过Southern杂交分析,得到了17个独立克隆.这17个克隆中至少含有13个不同的avrBs3/pthA家族基因.这些基因在GX1329基因组中有的单独存在,有的两个或两个以上串联存在.本工作基本上明确了菌株GX1329基因组中avrBs3/pthA家族基因的数量,为进一步研究菌株GX1329中avrBs3/pthA家族基因的功能奠定了基础.     

水稻细胞质雄性不育系和保持系的线粒体COⅠ、COⅡ基因组织结构差异的分析

我们研究了水稻珍汕97不育系和保持系的线粒体DNA的PstⅠ、HindⅢ、BamHⅠ酶切电泳带型,发现了不育系和保持系的线粒体DNA分子结构的显著不同,而不育系和保持系的叶绿体DNA的HindⅢ酶切片段没有差异。以线粒体的细胞色素C氧化酶亚基Ⅰ、Ⅱ(COⅠ、COⅡ)基因为探针,与珍汕97不育系、保持系的叶绿体、线粒体DNA酶切片段进行分子杂交,证明叶绿体DNA没有COⅠ、COⅡ基因的同源顺序,发现不育系和保持系COⅠ、COⅡ基因有组织结构上的差异,但不能确证这些差异是否与细胞质雄性不育有关。     

水稻组蛋白H_3基因的分离和组织结构分析

用噬菌斑原位杂交法,从一个水稻新品系的EMBL,基因文库中筛选到了两个组蛋白H,基因克隆,并制作了它们的限制性内切酶图谱,分析了它们的组织结构。其中λRH_3-1克隆包含2个组蛋白H_3基因区(分别为0.5kb和0.8kb的BamHI-EcoRI片段,两基因区距离5.8kb)。而λRH_3-2克隆只带有1个结构特异的H_3基因区(5.2kb的EcoRI-HindⅢ片段)。本文还就水稻组蛋白H_3基因在整个基因组中的组织结构特点进行了讨论。     

细胞质雄性不育高粱叶绿体 ndh D 基因的序列变异

片段SAAU－02 700特异地扩增自7种具可育细胞质的高粱材料的总DNA,含有叶绿体psa C(88bp)和ndh D(192bp)基因的部分序列。该片段与Eco Ri HindⅢ酶切的总DNA,线粒体DNA和叶绿体DNA杂交,在总DNA中获得了0.74kb的杂交带,而在叶绿体中获得0.74kb和0．45kb两条杂交带。与线粒体DNA无杂交;与经Hae Ⅲ酶切的总DNA杂交,在不育系中获得4．9kb的杂交带,而保持系的杂交带为4．45kb。参考GenBank中高粱的近缘物种玉米叶绿体基因组的序列,构建了ndh D基因区的酶切位点图谱,借此分析得出高粱不育系的叶绿体ndh D基因序列已发生改变。这种变异与高粱细胞质雄性不育反生的关系正在探讨中。     

棉铃虫核型多角体病毒p40基因序列分析

克隆了棉铃虫（Helicoverpa armigera）单粒包埋型核型多角体病毒（HaSNPV）基因组HindⅢ-H,J片段,其长度分别为10kb和6．7kb。通过对克隆片段末端测序,得到HaSNPV p40基因全序列。p40基因编码区全长966bp,预计可编码36.kD的多肽。HaSNPVp40基因核苷酸序列与HzSNPVp40基因有98％的相同性,这两种基因与BmNPVp40（GenBank-L33180）和AcMNPVgp41(GenBank-L22858)的在酸序列相似性43％左右,但四种蛋白对应于HzSNPVp40,HaSNPVp40的28－277氨基酸区域,同源性高达62％,并且存在一个保守的亲水性结构域,进一步将在基因组中相邻的Hind Ⅲ-H、J两片段用BamHⅠ、EcoRⅠ、HinⅢ、PstⅠ四种限制性切酶进行了分析。     

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.