Diversity of the ectomycorrhiza community at a uranium mining heap

Ectomycorrhiza (EM) community structure was analyzed at one bare heap site (BHS), one heap site with organic cover (HS-OH) and one reference site (RS) in the former uranium mining area near Ronneburg (Thuringia, Germany). Twenty-three EM morphotypes were distinguished, and 14 of them were additionally characterized by polymerase chain reaction (PCR), restriction fragment length polymorphism (RFLP) and internal transcribed spacer (ITS) sequence analysis. Colonization of birch by the different morphotypes was quantified, and the EM diversity at the different sites was investigated. Compared to RS, total EM colonization was reduced by 6% (P=0.851) at HS-OH and by 58% (P<0.001) at BHS. Likewise, EM diversity was reduced by 16% (P=0.229) at HS-OH and 52% (P<0.001) at BHS. The Sørensen similarity between EM samples from RS was nearly independent from the sampling date, whereas at HS-OH and especially BHS, the Sørensen similarity decreased with increasing time between the samplings. All EM fungal species dominating at the two heap sites were also present at RS. Thus, fungi with high tolerance against uranium and other stress factors at the heap sites (e.g. heavy metals, nutrient limitation, drought) were selected among all EM fungi of the area. Highly adapted fungi with a distribution restricted to the contaminated soils were not detected.Electronic Supplementary Material Supplementary material is available for this article at .     

Phylogenetic relationships in the North African genus <Emphasis Type="Italic">Hedysarum</Emphasis> as inferred from ITS sequences of nuclear ribosomal DNA

Sequences of the internal transcribed spacers (ITS) of nuclear ribosomal DNA were analysed to precise their length (637–643 bp) and resolve phylogenetic relationships among eight Mediterranean species of the genus Hedysarum (Fabaceae). The infra-specific variability levels of the ITS sequences of spontaneous population of H. coronarium proved a lack of polymorphism both in the length and in the sequences examined in this species. Hence, a consensus ITS sequence characterising each Hedysarum species has been investigated for analysis of inter-specific polymorphisms. The level of variation of ITS sequence was high enough to make the ITS1 and ITS2 a useful tool for phylogenetic reconstruction. However, ITS2 seems to be relatively more polymorphic and more informative than ITS1 regarding length or GC percent. The phylogenic relationships in the genus Hedysarum based on ITS1 and ITS2 sequences taken independently or together, are discussed in the context of current work in molecular biosystematics. Results exhibited the distinctiveness of the two H. spinosissimum subspecies (i.e. H. spinosissimum ssp. capitatum and H. spinosissimum ssp. spinosissimum). In addition, the great similarity of the ITS sequences between H. coronarium (the only cultivated species of the genus) and H. carnosum suggests the usefulness of the latter in selection programmes to improve pastoral production in semi-arid areas.     

黄杨属植物ITS序列分子进化特点分析

黄杨属植物种类多,分类较为混乱,特别是中国特有种珍珠黄杨的分类地位和系统进化存有争议,鉴于此,本文选取黄杨属5种植物和珍稀濒危植物珍珠黄杨的5个居群,对其rDNA ITS序列分析,结合GenBank中报道的黄杨属其它9个种,以板凳果属顶花板凳果的ITS序列为参照,用成对比较的方法对黄杨属18种植物的ITS序列比较发现,...     

有齿食道口线虫ITS及5.8S DNA片段的PCR扩增、克隆及序列分析

 运用PCR方法以保守引物NC5及NC2扩增了从广东阳江地区猪体分离的食道口线虫rDNA 的内转录间隔区（ITS）及5.8S序列。将PCR扩增出的片段纯化后克隆至pGEM-T Easy载体，用PCR技术及酶切鉴定阳性菌落，对阳性菌落质粒DNA进行测序。结果表明，扩增的片段大小为828 bp，包含部分的18S、28S及全部的ITS-1（362 bp）、5.8S(153 bp)及ITS-2（217 bp）序列。序列比较表明，该食道口线虫为有齿食道口线虫。本研究在国际上首次报道了中国猪有齿食道口线虫的ITS及5.8S序列，为食道口线虫的分子生物学的进一步研究奠定了基础。     

桑属ITS,TrnL-F,rps16序列分析及12个特殊桑资源评鉴

 【目的】分析桑属ITS,TrnL-F,rps16序列,探讨系统学价值,建立系统进化树,并对12个特殊桑资源进行评鉴,为开发利用提供理论依据。【方法】用67份桑资源,经DNA提取、PCR扩增、测序,测序结果用软件拼接、比对、除去非序列碱基,计算长度、G+C%含量、变异位点、信息位点、以构树、柘树为外类群,MP分析,根据分析结果,结合桑资源研究积累,对12个特殊桑资源进行评鉴。【结果】桑ITS(包括5.8S)基本序列576 bp ,变异范围576-590 bp,G+C% 59.55-62.25,40个信息位点;TrnL-F（包括tRNA-Leu内含子）基本序列923 bp , 变异范围920-924 bp, G+C%33.69-34.13,23个信息位点;rps16内含子基本序列929 bp ,变异范围923-929 bp,G+C%32.51-32.72,17个信息位点。最适碱基进化模型分别为(GTR+G),(GTR),(GTR+G),可能的分支概率,混合卡方概率值分别为0.000001 ,0.027175,0.000222, 三片段合并最适进化模型(GTR+G),基于模型,MP分析, 2538个位点,80个信息位点。分支图首先将黑桑M. nigra分出,其它桑种分成两支,第一支桑(mulberry),包括长穗桑、瑞穗桑、蒙桑、鬼桑、鸡桑、山桑、白桑、广东桑,第二支乔木桑(arbor),包括华桑、奶桑、川桑。12个特殊桑资源,神农华桑♀（优质木材桑）分在第二支,乔木特性,其它11个特殊桑资源分在第一支,具桑(mulberry)特性。【结论】桑属TrnL-F;rps16序列信息位点有限,单独研究桑属系统学,价值不大,与ITS序列合并,能增加分支图的信息位点,使分支图更具客观性,更近于似然。基于ITS,TrnL-F;rps16序列MP分支图,新疆黑桑为最原始类型,乌克兰等栽培种是进化类型。桑属可根据分支的亲缘关系,结合桑资源研究积累,评鉴特殊桑资源。     

腐烂茎线虫种内群体特异性检测研究

对腐烂茎线虫(Ditylenchus destructor Thorne)7个不同地理群体rDNA 中的ITS区进行PCR-RFLP和序列测定, 发现种内群体间无论在序列长度还是酶切图谱均存在一定的变异.根据其序列特征设计出种特异性引物, 对所供试的腐烂茎线虫群体及属间不同种群体样品的rDNA ITS区相应区段扩增以验证其特异性, 同时对分别由1～5条腐烂茎线虫成虫和幼虫提取的DNA进行特异性扩增以验证其灵敏度. 结果表明: 所有供试的腐烂茎线虫群体均可以特异性扩增一个346 bp的片段, 而所有供试的属外种均没有扩增片段;由1～5条腐烂茎线虫成虫和4～5条幼虫进行的特异性扩增也获得稳定的目的片段. 显示该特异性引物具有较高的稳定性、特异性和灵敏度, 能快速、准确地检测出腐烂茎线虫.     

3种鳢属鱼类ITS序列及系统进化分析

[目的]阐明乌鳢、斑鳢、月鳢核基因序列(ITS)的遗传变异情况,并用ITS序列研究鳢属系统进化关系.[方法]利用PCR扩增3种鳢的ITS,经克隆、拼接得到ITS全长后进行分析.[结果]乌鳢、斑鳢和月鳢ITS全序列长度为902、927、902或903bp.3种鳢ITS的G+C含量较高,约占72％.3种鳢之间核苷酸差异明显大于种内,明显的差异性ITS片段可以鉴别3种鳢.采用NJ和ML方法建立的进化树都表明乌鳢和斑鳢的遗传距离最近,而乌鳢和月鳢遗传距离最远.[结论]该研究为鳢属的分类鉴定、系统进化关系以及种间杂交提供基础资料.     

基于Smad 7基因为靶序列的RNA干涉作用研究

为了探讨小干涉RNA(siRNA)对靶基因Smad7mRNA表达的阻断作用,采用体外转录方法制备Smad7基因的小干涉RNAs(siRNAs),用脂质体介导瞬时转染BERP35T 2肺癌细胞系,并采用Northernblot杂交检测靶基因mRNA的表达丰度。结果表明,根据Smad7编码区序列在体外成功地制备了针对2个不同靶序列的siRNAs;Northernblot杂交显示,在转染siRNA的BERP35T 2细胞中,不管是内源性的还是外源性的,Smad7mRNA的表达丰度均明显下降。说明Smad7基因编码区中542563bp及701722bp2个区域均是siRNA作用的有效靶序列,本研究设计并制备的siRNAs能有效抑制Smad7基因的表达。     

Two- and three-domain bacterial laccase-like genes are present in drained peat soils

Laccases of fungal origin have been intensively studied due to their importance in various biotechnological applications. There is a constant demand for new laccases with improved properties such as stability at higher temperatures or at an alkaline pH. Growing molecular evidence suggests that laccases may also be widespread in bacteria. While only a handful of bacterial laccases have been purified and characterized, several novel traits have already been discovered (e.g. pH-stability and 2-domain organization of the enzyme as opposed to the usual 3-domain structure of fungal laccases). The aim of this study was to examine the diversity of bacterial laccase-like genes in two types of high-organic peat soil using a cloning and sequencing approach. Gene libraries prepared of small fragments (150 base pairs) revealed an amazing diversity of bacterial laccases. The fragments clustered in 11 major lineages, and one third of the 241 sequences resembled laccase-like genes of Acidobacteria. Additionally, a new primer was used to retrieve several larger fragments of the putative bacterial laccase genes that spanned all four copper-binding sites. Both “conventional” 3-domain laccases and the recently described 2-domain small laccases have been obtained using this approach, demonstrating the potential of the primer. The present study thus contributes to the understanding of the diversity of bacterial laccases and provides a new tool for finding laccase-like sequences in bacterial strains and soil samples.     

利用ITS序列和trnL-F序列探讨赖草属6个物种的系统发育关系

本研究克隆了赖草属6个物种Leymus karelinii,L.angustus,L.racemosus,L.arenarius,L.triticoides和L.ambiguus的ITS序列和trnL-F序列,并选用3个新麦属物种的ITS序列和27个小麦族二倍体物种的trnL-F序列,均以Bromus catharticus为外类群,用最大简约法构建系统发育树。结果发现:不同组的赖草属物种分别聚在一起;赖草属植物与新麦草属植物亲缘关系较近;新麦草属植物是这6个赖草属物种的母本来源。