云芝全基因组SSR位点分布及比较分析

本研究采用生物信息学方法统计分析了云芝基因组中SSR位点的数量、分布及频率等信息,对云芝基因组中含SSR的基因进行了功能注释,与其他5种伞菌纲真菌(双孢蘑菇、黑管孔菌、红缘拟层孔菌、金针菇和鲜红密孔菌)基因组进行了比较。结果发现云芝基因组中共有1 224个SSR位点,相对丰度为27个/Mb,三核苷酸重复类型分布最频繁。其中编码基因序列中包含299个SSR,仅次于基因间区,但是在非编译区、基因间区和内含子中SSR分布更加频繁。通过与nr数据库比对,485个含SSR的基因获得注释,其中115个基因通过GO注释到分子功能、生物进程及细胞组分3类中,108个基因通过KEGG注释到新陈代谢、遗传信息处理、细胞过程和环境信息处理4类中。设计58对引物主要用于物种鉴定,另外23对引物主要用于活性物质代谢研究和辅助育种。与其他伞菌纲真菌相比,云芝所含SSR的数量及相对丰度较低,并证明SSR数量与基因组大小无关,而某些特定的重复类型与GC含量有关。本研究为云芝的种群遗传学及进化研究奠定了理论基础。     

牦牛和水牛全基因组微卫星分布规律及其比较分析

微卫星(simple sequence repeats,SSRs)广泛分布于原核生物和真核生物基因组中,包括编码区和非编码区,是最常用的分子标记。本文利用生物信息学方法搜索和统计了牦牛和水牛全基因组中完整型SSRs序列,并对其生物信息学特征进行比较分析。牦牛和水牛全基因组中SSRs总数量分别为968 134个和1 052 443个,占其全基因组长度的比例分别为5.80‰和5.69‰。牦牛和水牛全基因组SSRs总丰度(366.01 vs 371.07个/Mb)和总密度(5 686.00 vs 5 799.34 bp/Mb)基本接近。牦牛和水牛全基因组SSRs丰富度分布模式如下:单核苷酸SSRs二核苷酸SSRs三核苷酸SSRs五核苷酸SSRs四核苷酸SSRs六核苷酸SSRs,这6种重复类型SSRs特征相互比较有显著差异,而相同重复类型SSRs特征基本一致。水牛第1条染色体上SSRs数量最多(72 934个),其次依次是第2、3、4条染色体,而较少的是第23、24条染色体,其所有染色体上SSRs丰度不存在显著差异(p0.05)。牦牛和水牛SSRs序列随着重复单元中核苷酸数量的增加,而其重复拷贝数逐渐下降。牦牛全基因组和水牛各染色体上各重复类型优势SSRs序列基本一致,并与普通牛、绵羊全基因组中不同重复类型SSRs优势序列相一致。     

马铃薯甲虫基因组SSR位点分析及引物效率的验证

SSR作为一种微卫星分子标记方法,被广泛用于遗传多样性相关的研究,而SSR引物是否高效是影响整个实验结果的重要因素。本研究利用生物信息学手段,对马铃薯甲虫基因组数据进行分析,搜索基因组SSR位点,并设计引物和验证SSR引物的效率。最终在马铃薯甲虫基因组中共检测出SSR位点81 937个,且这些位点中以单核苷酸、二核苷酸、三核苷酸重复为主;从SSR位点的平均分布距离来看,单、三、四核苷酸重复在马铃薯甲虫基因组中平均分布距离较小,分别为2.21 kb、8.39 kb、36.21 kb。马铃薯甲虫基因组SSR位点中,优势基序总数为68 388个,比例高达83.46%,其中,单核苷酸以A/T重复基序占绝对优势,二核苷酸重复的SSR位点中,以AG/CT重复基序为主,三核苷酸重复的位点中,以AAT/ATT重复基序为主;在ClassⅠ长度的SSR位点中,以二核苷酸、三核苷酸重复的数量最多,比例高达82.32%;最后设计19对SSR引物测试效率,其中14对成功扩增条带,11对为多态性引物,其平均多态性条带6.27条。多态性引物的PIC值在0.375-0.794,平均值为0.600,大于平均值的引物为6对,占有效扩增引物的54.5%。研究表明,利用生物信息发掘SSR位点的方法简便高效,这为后续进一步利用SSR引物研究马铃薯甲虫的遗传多样性,及在其它昆虫中开发SSR引物开发奠定研究基础。     

基于转录组数据的密花香薷SSR位点特征分析

利用MISA软件对密花香薷转录组42 362条Unigene进行SSR位点搜索,并对其SSR序列结构及分布特征进行了分析。结果表明:(1)密花香薷转录组Unigene序列中共检测到17 564个SSR重复序列,分布于11 903条Unigene上,出现频率为28.10%,平均每3 200 bp出现一个SSR位点。(2)单、二、三核苷酸重复类型为密花香薷转录组SSR位点的主导基序类型,占总SSR位点的97.27%,3种主导基序类型中,单核苷酸所形成基元类型数量最多,共检测到169个基元类型(51.22%),单核苷酸(A/T)n基元类型占明显优势,二核苷酸重复类型(AG/CT)n基元类型占优,分别占总SSR位点的50.60%和12.17%。(3)单核苷酸SSR位点所包含重复次数最多(49),重复次数介于10～66,同一基序类型不同重复次数所形成的SSR位点数量差异较大,随重复次数的增加,SSR位点数呈下降趋势。(4)密花香薷转录组二至六核苷酸基序SSR序列长度集中在12～30 bp区间,共包含有8 190个SSR位点,占所统计SSR位点的95.60%,1 589 (≥20 bp)个SSR序列具有极高的多态性,占所统计SSR位点的18.54%。综合出现频率、分布密度、基元重复次数和长度变异等多个研究结果发现,密花香薷转录组检索到的SSR序列表现出较高的多态性潜能,具有较大的开发价值。该研究为后续密花香薷SSR分子标记引物开发奠定了理论基础。     

中间锦鸡儿转录组EST-SSR标记系统性识别与引物筛选

旨在对中间锦鸡儿转录组数据库EST信息进行SSR系统性识别和初步验证,为进一步SSR分子标记开发提供依据。对Hi Seq2000测序技术获得的中间锦鸡儿转录组Unigenes进行SSR位点搜索,共获得45 706个SSR位点,出现频率为10.38%,平均4.30kb出现一个SSR位点。SSR重复类型以单核苷酸重复序列基元为主,所占比例为56.47%;二核苷酸、三核苷酸重复序列基元的数量所占比例分别是20.56%和21.04%;其他数量的基元所占比例仅为1.9%。多核苷酸重复类型中最多的为2核苷酸重复AG/CT;其次为3核苷酸重复AAG/CTT。针对EST-SSR位点随机挑选了150对引物,通过琼脂糖凝胶电泳进行PCR验证,其中有79对能获得扩增条带,21对引物扩增出单一条带,比例为14.0%。     

大蹄蝠全基因组微卫星分布特征分析

脊椎动物基因组含有丰富的微卫星信息。本研究对翼手目动物中的大蹄蝠全基因组及其基因的微卫星分布特征进行分析,并对含有微卫星编码序列的基因进行注释分析。结果表明,大蹄蝠全基因组大小为2.24 Gb,共含有497 883个微卫星,其中,数量和比例最多的是单碱基和二碱基重复类型,分别有173 953个(34.94%)和222 591个(44.71%),相对丰度分别为77.78 loci/Mb和99.52 loci/Mb。微卫星数量从单碱基重复到六碱基重复单元最多的类型分别为(A)n、(AC)n、(TAT)n、(TTTA)n、(AACAA)n和(TATCTA)n,比例分别为95.14%、55.25%、38.41%、22.17%、48.68%和20.30%。不同基因区和基因间区的数量及丰度不同,其中基因间区的微卫星数量及其丰度最大,分别为322 666个和2 541.57 loci/Mb,编码区的微卫星数量及其丰度最小,分别为1 461个和461.98 loci/Mb。基因间区和全基因组的微卫星的分布特征相似。编码区最多的微卫星类型为三碱基重复单元,外显子最多的微卫星类型为单碱基、二碱基和三碱基重...     

鲫鱼(Carassius auratus)表达序列标签资源的SSR构成与分布分析

分析鲫鱼EST资源的SSR信息,为开发EST-SSR标记奠定基础.从GenBank中获得鲫鱼EST序列,然后用Sequencher 4.8软件进行序列拼接得到Uni-EST序列,再通过SciRoKo 3.4软件扫描Uni-EST序列中的SSR,最后得出EST-SSR的分布特征、频率和重复基元类型等特征.通过搜索共获得9 230条鲫鱼EST原始序列,通过使用计算机软件进行预处理共得到全长为3.81×106 bp的无冗余Uni-EST 7 092条.在这些序列中共搜索出597个SSR位点,分布在545条Uni-EST序列中,发生频率为8.13％,EST-SSR的平均长度为(19.34±6.23) bp,平均每Mb含156.55个SSR位点.单核苷酸重复在鲫鱼EST-SSR中占主导地位,发生频率为39.53％,其次为二核苷酸重复,发生频率为36.68％以及三核苷酸重复的15.41％.在所有非单核苷酸重复基元中,AC基元出现频率最高,其次为AG.设计出引物404对.最后得出结论鲫鱼EST中SSR出现的频率较高,并且类型较为丰富,为进行遗传多样性分析和重要经济性状筛选等方面的研究提供了基础和指导.     

3种烟草基因组SSR位点信息分析和标记开发

利用生物信息学方法对绒毛状烟草、林烟草和本塞姆氏烟草3份烟草野生种基因组数据中的SSR位点信息进行了分析。结果表明,在全长分别为2.14Gb、2.44Gb和2.59Gb的绒毛状烟草、林烟草和本塞姆氏烟草基因组中分别获得153 357个、196 493个和278 784个SSR位点,平均相隔13.94kb、12.42kb和9.31kb出现一个SSR。在SSR位点分布区域上,绝大部分的SSR位点分布在内含子和UTR(尤其是5′-UTR)区域;在SSR基序类型上,主要集中在二、三碱基基序且二者占总SSR位点数目的80%以上,并以二碱基基序类型丰度最高;在SSR基序结构上,基因组中出现频率及数量最高的是含有A(T)n的基序结构;在SSR基序的重复次数上,除单碱基基序类型外,重复次数多在3~10次之间。利用分别属于5个不同烟草组的8份烟草材料验证所合成的300对引物,所有合成的引物均能扩增获得目标片段,其中有80对引物存在扩增多态性。表明来源于绒毛状烟草、林烟草和本塞姆氏烟草基因组的SSR标记在亲缘关系相对较近的烟草种间具有高度保守性和通用性,基于此3份烟草野生种基因组数据开发SSR引物用于后续的相关遗传研究具有可行性。     

濒危植物羊踯躅全基因组SSR标记开发与鉴定研究（英文）

该研究利用基于全基因组限制性酶切位点简化基因组测序技术(RAD-seq技术),开发濒危植物羊踯躅(Rhododendron molle G.Don)全基因组SSR标记,并对3个群体共63份羊踯躅材料进行验证鉴定,为进一步研究羊踯躅的遗传多样性和群体遗传结构以及保护利用提供技术支持。结果显示:(1)羊踯躅基因组测序获得原始数据7.653Gbp,过滤后为7.513Gbp;经组装发现,羊踯躅171.534Mbp的基因组分布在498 252contigs中。(2)通过SSR检测,在11 961SSR位点中获得了11 687对SSR分子标记,并且二核苷酸为基序的重复类型最丰富,达51.76%。(3)随机选取128对SSR标记在6个羊踯躅株系中进行PCR扩增,获得20对高多态性的SSR标记。(4)用所选的20对多态性SSR标记对3个群体共63份羊踯躅材料进行验证分析发现,这些多态性SSR标记位点的等位基因数为4~16个,期望杂合度(He)为0.489~0.908。研究表明,羊踯躅的SSR丰度适中,且二核苷酸为羊踯躅中最丰富的重复序列,该实验进一步证明RAD-seq技术是一种经济有效的基因测序方法,实验中开发的SSR引物将有助于进一步研究羊踯躅和其他近缘种的群体结构和多样性。     

禾本科植物微卫星序列的特征分析和比较

微卫星或简单序列重复(simple sequence repeats,SSRs)广泛存在于真核生物的基因组中,是目前最有用的分子标记之一.本研究首次利用全基因序列对4个禾本科植物种,包括二穗短柄草(Brachypodium distachyon)、水稻(Oryza sativa)、高粱(Sorghum bicolor)和玉米(Zea mays)的SSR的类型、丰度(数量)、相对丰度、频率、长度和偏好性等进行了分析和比较.结果表明,禾本科植物中SSR的丰度与基因组大小成正比,而相对丰度与基因组大小相关不显著;三核苷酸和六核苷酸基序类型比其它类型更为丰富;不同物种皆呈现出明显的基序偏好性.总的来讲,除个别例外,禾本科植物的基因组中更倾向于富含A/T的基序.本研究结果为研究禾本科植物的基因组进化和SSR标记的开发提供了有价值的信息.     

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.     

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.     

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.     

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.     

The mechanism of hatching of eggs of Haemonchus contortus

Fluid collected from hatching eggs of Haemonchus contortus contained a lipase which hydrolysed 2-naphthyl laurate (about 0·7 μmol naphthol freed /h/10⁶ eggs). The fluid also hydrolysed l-leucinamide (about 2·3 μmol leucine freed/h/10⁶ eggs). The fluid when added to normal or heated eggs caused ‘hatching’. ‘Hatching’ also occurred in exsheathing fluid from infective juveniles and in a preparation of pancreatic lipase containing leucine aminopeptidase. A purified mammalian leucine aminopeptidase in combination with several different lipases did not attack egg shells.The ‘spontaneous’ hatching of eggs of H. contortus was strongly inhibited by 1,10-phenanthroline, 10^?3M, and this inhibition was reversed by Zn²⁺. However, the inhibition of ‘hatching’ of eggs in externally applied hatching fluid, or the hydrolysis of leucinamide in hatching fluid was generally less marked.