基于nrDNA ITS序列的18份宁夏枸杞资源的遗传多样性(英文)

[Objective] The study aimed to investigate the genetic polymorphism of eighteen Lycium barbarum resources via nrDNA ITS sequencing. [Method] The genomic DNAs from Lycium barbarum leaves were isolated by modified CTAB method for PCR amplification on the nrDNA ITS region using specifically synthesized primers; the amplified fragments were cloned and sequenced, then the sequencing results were clustered. [Result] nrDNA ITS sequences of the tested eighteen Lycium barbarum were firstly obtained in the present study. For all eighteen tested materials, the variation range of whole ITS region was 559-634 bp, with an average of 612 bp; alignment analyses showed that the whole length of internal transcribed spacer (ITS1+ITS2) was 480 bp, within which there are 194 variation sites (accounting for 40.4%) and 286 conserved sites (accounting for 59.6%). The cluster results showed that the eighteen tested materials could be grouped into three classes. [Conclusion] Analysis of nrDNA ITS sequence may avail to identify the Lycium barbarum germplasm resources.     

我国部分区域链格孢属rDNA ITS区序列分析(英文)

[Objective] The study aimed to identify Alternaria Nees. from some areas of China at molecular level by analyzing the rDNA ITS sequence.[Method] The DNA sequences coding for the 5.8S rDNA and the flanking internal transcribed spacers (ITS1 and ITS2) were amplified by PCR with universal primers ITS4 and ITS5 and subsequently sequenced for 34 Alternaria isolates from different areas of China.[Result] Sequences analysis showed that 5.8S rDNA was 159 bp and no variation in tested 34 isolates. There had variables sites in ITS. The isolates that had same sequences as A.tenuissima or A.alternata all put up eurytopicity to area and host. The variables sites of the isolates showed the diversity of Alternaria in the hosts of Oleaceae, Rosaceae and Solanaceae. At the same time that ITS could not clearly separated the isolates was indicated. The results indicated that the phylogenetic relationship were not closely related to the geographical origin and hosts of these isolates.[Conclusion] The sequence analysis of ITS region could provide theory basis for the identification of Alternaria Nees..     

枸杞nrDNA ITS测序鉴定的初步研究（英文）

[Objective] The study aimed to identify wolfberry(Lycium Linn.)germplasm resources at molecular level by analyzing the nrDNA ITS sequence.[Method] Genomic DNA from wolfberry leaves extracted by modified CTAB method were regarded as templates for PCR amplification by specific primer,clone and sequencing.[Result] The nrDNA ITS sequences were obtained and then differentiated among three tested materials.[Conclusion] PCR amplification and sequencing on nrDNA ITS is a feasible approach to identify different wolfberry germplasm resources.     

西川红景天nrDNA ITS序列初步研究

[目的]对西川红号天nrDNA ITS序列进行分析,并与cpDNA（叶绿体DNA）trnS-trnG序列和rpl20-rps12序列进行比较,从而初步比较2套植物基因组的进化速率。[方法]采用改良CTAB法从硅胶干燥的西川红景天叶片中提取总DNA,并对nrDNA ITS区进行扩增、纯化、测序,然后与cpDNA trnS-trnG和rp120-rps 12序列进行比较。[结果]序列比对后得到长度为701bp的ITS序列,其中变异位点13处,占总序列的1.85％。在13处变异位点中,8处为碱基置换,5处为插入／缺失。（A＋T）含量为46．9％,（G＋C）含量为53．1％。核苷酸多样性为0．00427。[结论]西川红景天nrDNA ITS区域较cpDNA trnS-trnG序列和rp120-rps12序列保守,进化速率较慢。     

一种鉴定菜用枸杞的新方法——nrDNA ITS测序法(英文)

[Objective] The study aimed to identify Lycium Linn. at molecular level.[Method] The nrDNA ITS sequence of 5 edible Lycium Linn. germplasm resources were investigated. [Result] The nrDNA ITS regions of five edible Lycium Linn. germplasm resources were sequenced. The whole sequences varied from 628 bp to 632 bp,with the average length of 630 bp. Total 79 variation sites were observed in the sequences,which accounts for 12.5%. [Conclusion] Sequence analysis based on nrDNA sequencing provides a new approach to identify edible Lycium Linn. germplasm resources.     

ISSR Marker and ITS Sequence Study of Melampsora Larici-populina

To compare the differences in intertranslation space of ribosomal DNA （ITS） of Melampsora larici-populina, between the isolates from China and isolates from other countries, this study investigated ITS sequences and ITS polygenetic tree based on 11 isolates that were collected from 5 races in different parts of China. The results indicated that there was no difference among the ITS sequences of 11 isolates from China. The ITS sequence of isolates from China was more homogeneous with that of isolates from Britain compared with France, Germany, and Canada. Intersimple sequence repeat （ISSR） markers were also used to study the genetic division of Melampsora larici-populina, and the results showed that the 11 tested isolates could be divided into Western population and Northern population. Genetic diversity index of race C2 was significantly different from that of races C4, C3, and C1, and no significant differences were observed among the other races. Pathogenicity division of races must not harmonize with their genetic division, except race C2. The ITS region is conservative, and ITS sequence is not fit for studying the differences that existed among the races. ISSR marker can be used for intraspecies population study, and Melampsora larici-populina in China can be divided into two populations.     

Identification of Powdery Mildew Pathogen and Ribosomal DNA-ITS Sequence Analysis on Melon

Identification of powdery mildew pathogens on melon（Cucumis melo） is important for melon breeding and diseaseresistant germplasm selection. In this study, a powdery mildew pathogen that infected melon plants in Heilongjiang Province, China, was investigated in terms of host identification, morphological characteristics and phylogenetic relationships. The morphological characteristics of the pathogen were observed at five phases in the life cycle： germinating conidia, primary germ tube, hyphae, conidiophores, and colonization. The conidia were elliptical, colorless, catenulate, and the average length was 29.07 μm and average width was 17.82 μm. One ascus and eight ascospores were produced. DNA was extracted from 0.01 g conidiophores from a strain of powdery mildew pathogen that infected melon. ITS ribosomal DNA region（524 bp） was amplified with the universal ITS1 and ITS4 primers. The nucleotide sequence showed 100% similarity with ITS sequences for three Podosphaera fusca strains obtained from the GenBank database. The identity of the pathogen was confirmed as Sphaerotheca fuliginea. International standard differential hosts were used to identify S. fuliginea strain as 2F race. These results supported the notion that Podosphaera fusca was a synonym of S. fuliginea.     

小球藻的分离及其DNA提取方法的研究(英文)

[Objective] The aim of this study is to isolate Chlorella vulgaris(chlorella)and extract its genomic DNA.[Method] Both the dilution method and drip method were employed to isolate chlorella from lake water samples;the conditions for culturing chlorella were optimized and its genomic DNA was extracted by improved CTAB method and SDS method.[Result] The proper conditions for chlorella culture were as following:temperature 20-25 ℃,illumination 4.39-5.86 W/m2 and rotational speed 100-150r/min;improved CTAB method was suitable for extracting genomic DNA from chlorella.[Conclusion] The study is helpful to study the chlorella at molecular level and promote the exploitation and utilization of chlorella resources.     

小球藻的分离及其DNA提取方法的研究(英文)

[Objective] The aim of this study is to isolate Chlorella vulgaris(chlorella)and extract its genomic DNA.[Method] Both the dilution method and drip method were employed to isolate chlorella from lake water samples;the conditions for culturing chlorella were optimized and its genomic DNA was extracted by improved CTAB method and SDS method.[Result] The proper conditions for chlorella culture were as following:temperature 20-25 ℃,illumination 4.39-5.86 W/m2 and rotational speed 100-150r/min;improved CTAB method was suitable for extracting genomic DNA from chlorella.[Conclusion] The study is helpful to study the chlorella at molecular level and promote the exploitation and utilization of chlorella resources.     

Isolation of Chlorella vulgaris and Its DNA Extraction Methods

[Objective] The aim of this study is to isolate Chlorella vulgaris(chlorella)and extract its genomic DNA.[Method] Both the dilution method and drip method were employed to isolate chlorella from lake water samples;the conditions for culturing chlorella were optimized and its genomic DNA was extracted by improved CTAB method and SDS method.[Result] The proper conditions for chlorella culture were as following:temperature 20-25 ℃,illumination 4.39-5.86 W/m2 and rotational speed 100-150r/min;improved CTAB method was suitable for extracting genomic DNA from chlorella.[Conclusion] The study is helpful to study the chlorella at molecular level and promote the exploitation and utilization of chlorella resources.     

苞叶姜的nrDNA ITS2和cpDNA psbB-H序列 分子进化特点的分析

采用改良的CTAB法从硅胶干燥的苞叶姜(Pyrgophyllum yunnanense)叶片中提取总DNA,对nr DNA ITS2和cp DNA psb B-H区域进行PCR扩增、测序和序列分析,初步研究2套植物基因组的变异速率。nr DNA ITS2序列长224 bp,有变异位点3处,变异位点百分率为1.339%,(G+C)含量为60.7%。cp DNA psb B-H序列长623~625 bp,有2个碱基插入缺失,变异位点8处,变异位点百分率1.284%,(G+C)含量为33.9%,cp DNA核苷酸多态性(0.00551)比nr DNA(0.00295)高。苞叶姜的这2个片段,变异速率相近,遗传分化指数相同(Fst=1.000),居群间高度分化。ITS2序列和psb B-H序列单倍型中性检验结果一致,表明苞叶姜居群处于长期稳定状态。psb B-H错配分布(Mismatch-distribution)分析,表示苞叶姜的现有分布范围近期可能经历了居群扩张。因此,苞叶姜的谱系地理学研究可结合nr DNA ITS2序列和cp DNA psb B-H序列来分析。     

中药秦艽基原植物的DNA分子鉴定

[目的]分析秦艽基原植物间不同DNA序列的差异,为秦艽药材DNA条形码的筛选和基原鉴定提供分子证据。[方法]采用PCR扩增纯化后直接测序的方法,测定大叶秦艽G.macrophylla pall.、麻花秦艽G.straminea Maxim.、粗茎秦艽G.crassicaulis Duth-ieex Burk.、小秦艽G.dahurica Fisch、黄管秦艽G.officinalis H.Smith5种植物的核糖体DNAITS、叶绿体DNA psbA-trnH核苷酸序列,并作序列同源性分析。[结果]cpDNA psbA-trnH序列长度变异范围为316-318bp,有7种不同的单倍型,单倍型间有7个变异位点,序列的GC含量为21.2%。最大简约树的聚类结果与单倍型反映的结果一致。nrDNA ITS序列长度变异范围为624～625bp。有5种不同的单倍型、单倍型间有12个变异位点,序列的GC含量为59.3%。最大简约树的聚类结果表明,小秦艽与麻花艽聚为一支,大叶秦艽与黄管秦艽聚为一支,粗茎秦艽位于聚类图的最基部。[结论]nrDNAITS序列较适合作秦艽基原植物的DNA分子鉴定。     

枸杞nrDNA ITS测序鉴定的初步研究

ITS区序列指DNA基因内的转录间隔区序列,包括ITSl、ITS2及5．8S3个区段。ITS区序列测定分析能够鉴别同属植物相似种以及药用植物混淆种。笔者以枸杞属内不同种间ITS序列为切入点,对枸杞nrDNA ITS序列进行分析研究,为从分子水平鉴定枸杞品种提供参考。     

一种鉴定菜用枸杞的新方法——nrDNA ITS测序法

笔者以5种菜用枸杞为试验材料,开展nrDNA ITS序列分析研究,探索适宜于枸杞nrDNA ITS序列分析的优化技术体系,从而为建立分子水平的菜用枸杞鉴定标准提供参考。     

利用ITS序列探讨锦鸡儿属(Caragana Fabr.)植物系统关系(英文)

[目的]利用ITS序列探讨锦鸡儿属(Caragana Fabr.)植物系统关系。[方法]以锦鸡儿属11个系29种为代表材料,选择性扩增nrITS序列并双向测序,结合黄耆亚族Astralinae(Adens)Benth其他6属7个代表种的nrITS序列进行最大简约性(MP)和最小进化(ME)的系统发育分析。[结果]锦鸡儿植物ITS序列长度在611-614bp之间,与外类群排序后长度为655bp,共有170个可变位点,其中107个简约信息位点,简约信息位点在总排序序列中达16.3%,可以为属内及属间系统关系提供有力的分子证据;锦鸡儿属在系统发育上不是一个单系类群,与丽豆属(Calophaca Fish.exDC.)植物具有极为相近的亲缘关系;Sect.tragacanthoides的种在MP和ME进化树中位置分散,其组的分类有待进一步研究。卷叶锦鸡儿(C.ordosica,新种)虽然形态上与垫状锦鸡儿(C.tibetica)相似,但它与荒漠锦鸡儿(C.roborovskyi)遗传学关系紧密;C.davazamcii是一个独立的种。[结论]ITS序列在锦鸡儿属内及属间的系统学研究中具有重要的参考价值。     

枸杞nrDNA ITS测序鉴定的初步研究

[目的]对枸杞nrDNA ITS(核糖体DNA基因内转录间隔区)序列进行分析,从而在分子水平对枸杞做出鉴定。[方法]采用改良CTAB法提取枸杞叶片DNA,利用合成的特异引物对其DNA中nrDNA ITS区进行扩增、克隆,对目的片段测序分析。[结果]克隆到枸杞nrDNA ITS片段并获得其碱基序列,成功找到3个供试枸杞种质材料的nrDNA ITS序列差异。[结论]枸杞nrDNAITS区的扩增和测序可以在分子水平对枸杞不同种质进行鉴别。