白花柽柳nrDNA的ITS序列片段研究

运用PCR直接测序法 ,对白花柽柳 (TamarixalbiflonumM .T .Liu .)的nrDNA的ITS区 (包括ITS -l,5 .8SrDNA和ITS - 2 )进行序列测定。结果表明 ,该植物的ITS序列和长穗柽柳 (Tamarixelongata)的序列差别不大 :ITS - 1片段的长度都是 2 5 9bp ;ITS - 2片段的长度同为 2 44bp ;G C百分含量仅相差 1% ;碱基差异共发生 18处。由ITS及 5 .8SrDNA序列分子证据看 ,白花柽柳更可能是长穗柽柳的变异。     

Development of specific PCR primers for identification and detection of <Emphasis Type="Italic">Phytophthora capsici</Emphasis> Leon

A PCR-based method was developed for the identification and detection of Phytophthora capsici in pepper plants. Three PCR primers (CAPFW, CAPRV1 and CAPRV2) specific for P. capsiciwere designed based on the sequence of its internal transcribed spacer regions. CAPFW/CAPRV1 amplify a 452 bp product from P. capsici DNA whereas CAPFW/CAPRV2 a 595 bp fragment; neither set amplifies DNA from pepper or several fungi pathogenic to pepper. In conventional (single-round) PCR, the limit of detection was 5 pg DNA for both primer sets, whereas in nested PCR the detection limit for both was of 0.5 fg. However, when the dilution series of target DNA were spiked with plant DNA, amplification declined two-fold in both conventional and nested PCR. The CAPFW/CAPRV2 set in conventional PCR was used to detect P. capsici DNA in inoculated plants. Detection occurred as soon as 8h post-inoculation in stem samples from infected but still symptomless plants. The method was also tested to detect fungal DNA in infected soils.     

西藏自治区首次发现螟黄赤眼蜂

为发掘西藏地区本土的赤眼蜂资源,本研究利用灭活米蛾卵卡在西藏林芝嘎玛地区的梨园采集到一个赤眼蜂种群。采用ITS2序列比对鉴定所采集的赤眼蜂种类,并测定该蜂的生物学指标。结果表明：西藏林芝嘎玛地区所诱集的赤眼蜂鉴定为螟黄赤眼蜂Trichogramma chilonis Ishii,羽化率为80%,雌性比为57.14%,耐饥饿时间为1.08 d,后足径节长度为157.6μm。本研究将为发掘西藏地区赤眼蜂资源提供参考,并为利用赤眼蜂在西藏自治区开展生物防治实践奠定基础。     

杂草稻nrDNA ITS片段的PCR扩增条件优化及测定

对杂草稻的nrDNA ITS片段的PCR扩增条件进行了优化并测定,建立的PCR最优体系为：20μL反应体系含1μL模板DNA,0．125mmoL／L dNTPs,0．5μmol／L正反向引物,1U Taq酶,2．0μL 10&#215;Taq PCR Bufter;退火温度为57℃。这样的条件下充分保证了ITS PCR产物的质量和纯度要求,直接测序结果为600bp左右,与网上结果十分类似,表明结果准确可靠。这些ITS片段的系统学信息将为杂草稻的起源进化提供有力的分子水平证据。     

基于ITS和形态学系统分析新疆桦木属种间系统发育关系

以新疆桦木属的6个种和其他10种桦木属植物为材料,利用Clustal Omega和MEGA 7.0软件对ITS基因序列进行遗传分析,同时利用SPSS 25.0软件对7个形态性状进行主成分和聚类分析,探讨新疆桦木属植物的系统发育关系。结果表明：在ITS遗传演化分析中,新疆桦木属中的桦木亚组和柴桦亚组遗传距离较近,为0.0047,桦木亚组内部绝对遗传距离为0.0060;在形态性状主成分分析中,特征值大于1的累计贡献率为84.5%,桦木属植物分类性状主要取决于生活型、叶片形状、果苞背部被毛、树皮剥裂及纹路;新疆桦木属植物ITS序列和形态学聚类结果基本一致,盐桦与甸生桦聚为一组,与《中国植物志》中盐桦归属于桦木亚组,而甸生桦属于柴桦亚组的分类存在一定差异,但与《新疆植物志》中两者被划分在柴桦亚组的结论有相似之处。     

通过核基因ITS2片段研究四种鲇形目鱼类进化

采用鲇形目鱼鱼类通用引物扩增了鲇形目鱼类4种鱼,长吻(Leiocassi longirostris)、南方南方大口鲇(Si-lurus meridionalis)、黄颡鱼(Pseudobagrus fulvidraco)、斑点叉尾(Ictalurus punctatus)核DNA的ITS2片段,并构建UPMGA,NJ,ME和MP系统树。结果显示:根据遗传距离,斑点叉尾和黄颡鱼的亲缘关系最近(D=0.0661),接下来是黄颡鱼和南方大口鲇(D=0.1100),南方大口鲇和长吻(D=0.1184),斑点叉尾和南方大口鲇(D=0.1266),黄颡鱼和长吻(D=0.1490),斑点叉尾和长吻的亲缘关系是最远的(D=0.1503)。结果表明:长吻最早分化,其次是南方南方大口鲇和黄颡鱼,而斑点叉尾分化最晚。     

江苏省小麦禾谷孢囊线虫群体核糖体DNA-ITS区序列分析

对采集自江苏省徐州、宿迁、连云港和盐城4个地区的15个小麦禾谷孢囊线虫群体进行核糖体DNA-ITS区的RFLP分析。结果表明:测定的所有群体均具有相同的酶切图谱,既具有与国外B型群体(印度群体)相同的AluⅠ和RsaⅠ酶切图谱,同时也具有中国群体独特的HinfⅠ和Tru9Ⅰ酶切图谱(C型)。采用Neighbor-Joining法(MEGA4.0)构建的ITS序列系统进化树显示:所有15个江苏群体均聚在小麦禾谷孢囊线虫组(Heterodera avenae group)下的小麦禾谷孢囊线虫复合种群(H.avenae complex)分支内,且多数江苏群体与草地孢囊线虫(H.pratensis)德国和俄罗斯群体的遗传距离相对较近。通过江苏群体与其他近源群体的ITS序列分析比较,河南郑州群体与澳州孢囊线虫(H.australis)的遗传距离较近,青海群体和河南郑州群体在分子遗传水平上具有明显的地域性,而江苏省和我国其他省份的小麦孢囊线虫群体具有丰富的遗传多样性。     

DNA条形码基因ITS·ITS2及rbcL在苍耳属可采用相同的PCR条件（英文）

[目的]为植物DNA条形码标准基因筛选研究提供参考,减少植物DNA条形码研究中的工作量。[方法]对7种苍耳属植物ITS、ITS2及rbcL基因采用相同的扩增条件（95℃4 min;[35 cycles：94℃30 s;52℃45 s;72℃45 s];72℃10 min;4℃保存）。[结果]3种DNA条形码基因同时成功扩增。[结论]这说明植物DNA条形码基因中ITS、ITS2及rbcL的PCR条件存在合并可能性。     

ITS—RFLP分析进行昆虫病原线虫斯氏属和异小杆属的分类鉴定

本研究通过PCR扩增和六种限制性内切酶（AluⅠ，HinfⅠ，MboⅠ，RsaⅠ，HaeⅢ和PvuⅡ）酶切，对国内害虫防治上常用的几种昆虫病原线虫，包括斯氏属S.car-pocapsae,S.feltiae和S.glaseri以及异小杆属H.bacteriophora,H.zealandica,H.indicus和H.megidis等8个品系rDNA-ITS进行分析。建立起可以区分各线虫种的标准RFLP图谱。该方法快速简便，稳定可靠，需要的样品量少。可以用于新鲜的，或冻存的样品，甚至分析单条的线虫，不仅可进行昆虫病原线虫的快速分类鉴定。而且进一步可以应用于线虫田间释放的辅助监测。实际田间感染率的测定和线虫毒力的比较。     

Influence of repeated prescribed burning on the soil fungal community in an eastern Australian wet sclerophyll forest

A long-term prescribed burning experiment, incorporating replicated plots that receive burning biennially (2 yr burn) or quadrennially (4 yr burn) and unburned controls, has been maintained in a wet sclerophyll forest at Peachester, Queensland, Australia since 1972. In 2003 we extracted DNA from soil collected from the experimental plots and investigated the influence of the burning on the soil fungal community by comparing denaturing gradient gel electrophoresis (DGGE) profiles of PCR-amplified partial rDNA internal transcribed spacer regions (ITS1). Canonical analysis of principal coordinates (CAP) of the DGGE profiles of the upper 10 cm of the soil profile grouped the data strongly according to treatment, indicating that both burning regimes significantly altered fungal community structure compared to the unburned controls. In contrast, no obvious trend was observed for soil from a depth of 10-20 cm of the profile. Sequencing of selected DGGE bands found no obvious patterns of presence/absence of taxonomic groups between the treatments. Analysis of soil nitrogen and carbon by mass spectrometry indicated that total soil C and N, along with both gross and net N mineralisation, were significantly lower in 2 yr plots compared to control and 4 yr plots.