山姜属中药草豆蔻和益智nrDNA ITS区序列的测定

中药草豆蔻、益智的原植物分别为山姜属草豆蔻（Ａｌｐｉｎｉａ ｈａｉｎａｎｅｎｓｉｓ Ｋ．Ｓｃｈｕｍ．）与益智（Ａ．ｏｘｙｐｈｙｌａ Ｍｉｑ．）。本文采用ＰＣＲ直接测序法,首次测定了它们的核糖体ＤＮＡ ＩＴＳ区序列。结果显示,两者序列长度（ＩＴＳ１＋ＩＴＳ２）分别为４０３ｂｐ与４０４ｂｐ,序列间具有２７个变异位点（包括５．８Ｓ编码区）。本研究为山姜属中药材的ＤＮＡ分子鉴定提供了必要的序列资料。     

ITS（nrDNA）片段在冷杉属植物中的长度多态性及其在松科的系统与 …

由ＰＣＲ反应扩增了２８种冷杉属（ＡｂｉｅｓＭｉｌｌ．）植物的ｎｒＤＮＡ的ＩＴＳ（包括ＩＴＳ１、ＩＴＳ２和５．８ｒＤＮＡ）片段,经过与１００ｂｐ和１ｋｂ的标准分子量ＤＮＡ进行比较和某些类群ＩＴＳ的全序列和部分序列测定,发现分布于墨西哥和２种分布于北美的冷杉属植物的ＩＴＳ片段长度约为２５００ｂｐ,而分布于欧亚大陆和其他分布于北美的冷杉植物的ＩＴＳ长度约为１７００ｂｐ,二者相差约８００ｂｐ。Ａ．ｂｒａｃ     

山羊草属二倍体物种核rDNA ITS区序列及其系统发育关系分析

测定了山羊草属（Ａｅｇｉｌｏｐｓ）二倍物种核ｒＤＮＡ ＩＴＳ区序列,发现其碱基粉介于６０１－６０７之间,比报道的小ｒｉｕｃｅａｅ）其他属的ＩＴＳ区我略长,Ｇ＋Ｃ含量达６１．１％～６２．９％,序列间的分化距离为０．００５０～０．０４６８。用ＰＨＹＬＩＰ３．５ｅ软件包对所测得的ＤＮＡ数据进行聚类分析,结果显示：１．Ａｅ．ｓｐｅｌｔｏｉｄｅｓ与该组其他种相距很远,支持将其从Ｓｉｔｏｐｓｉｓ组中独立出来,     

一个新的水稻MADS—box基因的克隆及表达分析

根据ＭＡＤＳ－ｂｏｘ基因保守区结构,设计简并性引物,利用３＇ＲＡＣＥ从水稻（Ｏｒｙｚａ ｓａｔｉｖａ Ｌ．）中克隆了１个新的水稻ＭＡＤＳ－ｂｏｘ基因的ｃＤＮＡ片段,同时利用５＆ＲＡＣＥ获得了全长ｃＤｎＡ命名为ＦＤＲＭＡＤＳＳ。序列分析表明,该ｃＤＮＡ全长１４０６ｂｐ,开放阅读框共编码２３３个到,具有典型的植物ＭＡＤＳ－ｂｏｘ基因的结构。推测的氨基酸序列与拟南芥的ＭＡＤＳ－ｂｏｘ基因,ＡＧＬ１４同     

采用PCR—RFLP和RAPD对球壳孢目真菌系统学的研究

对球壳孢目真菌首次采用ＰＣＲ－ＲＦＬＰ和ＲＡＰＤ进行了系统发育研究,以ＩＴＳ１和ＩＴＳ４为引物对４属１２种２４个菌株的核糖体ＤＮＡ转录间区进行了ＰＣＲ扩增,４种内切酶酶切,结果表明：主要属的ＩＴＳ区长度不同,同属不同种相同。Ｃｏｎｉｏｔｈｙｒｉｕｎ,Ｐｈｙｌｌｏｓｔｉｃｔａ,Ａｓｃｏｃｈｙｔａ,ＳｅｔｏｒｉａＩＴＳ区长度分别为６３０,５６０,５５０,５４０ｂｐ;酶切图各间差别明显,属内种间基本一致     

普通小麦基因组最可能的4个供体的ITS1和ITS2序列及其亲缘关系

对普通小麦（ＴｒｉｔｉｃｕｍａｅｓｔｉｖｕｍＬ．）基因组（ＡＡＢＢＤＤ）最可能的供体－Ｔ．ｕｒａｔｒｔｕＴｈｕｍ．（ＡＡ）、Ｔ．ｍｏｎｏｃｃｕｍｖａｒ．ｂｏｅｏｔｉｃｕｍ（Ｂｏｉｓｓ．）ＭＫ（ＡＡ）、ＡｅｇｉｌｏｐｓｓｐｅｌｔｏｉｄｅｓＴａｕｓｃｈ．和Ａｅ．ｔａｕｓｃｈｉｉ（Ｃｏｓｓ．（ＤＤ）的核糖体ＲＮＡ基因ＩＴＳ区进行了ＰＣＲ扩增和克隆,并测定了ＩＴＳ１和ＩＴＳ２的ＤＮＡ序列,讨论和纠正了前人     

谷子核酮糖1,5—二磷酸羧化加氧酶大亚基基因的核苷酸序列

谷子（Ｓｅｔａｒｉａ ｉｔａｌｉｃａ）叶绿体ＤＮＡ 含有ｒｂｃＬ基因的３．０ ｋｂ Ｂｇｌ Ⅱ＋ ＸｂａⅠ酶切片段被克隆到ｐＢｌｕｅｓｃｒｉｐｔ ＳＫ （－ ）质粒载体上,并构建了该基因的限制性酶切图谱,测定了该基因的１９９０ ｂｐ 全序列。其中基因的编码区长１４３１ ｂｐ,共编码４７６ 个氨基酸,推测其分子量为５２６７９ Ｄ。其３８９ ｂｐ 的５′上游区含有类似于原核生物的－ １０ ｂｏｘ、－ ３５ ｂｏｘ 和ＳＤ序列。其１７０ ｂｐ ３′下游区含有３ 个茎环结构。对Ｃ４ 植物和Ｃ３ 植物中的ｒｂｃＬ基因序列进行比较,表明在编码区、启动区和３′下游区没有差别。由此认为Ｃ４ 植物中ｒｂｃＬ基因的细胞特异性表达与其序列无关     

采用PCR-RFLP和RAPD对球壳孢目真菌系统学的研究

对球壳孢目真菌首次采用ＰＣＲ－ＲＦＬＰ和ＲＡＰＤ进行了系统发育研究。以ＩＴＳ１和ＩＴＳ４为引物对４属１２种２４个菌株的核糖体ＤＮＡ转录间区（ＩＴＳ）进行了ＰＣＲ扩增,４种内切酶（ＡｌｕＩ,Ｈｈａ,ＭｓｐＩ,ＴａｑＩ）酶切,结果表明：主要属的ＩＴＳ区长度不同,同属不同种相同。Ｃｏｎｉｏｔｈｙｒｉｕｍ,Ｐｈｙｌｏｓｔｉｃｔａ,Ａｓｃｏｃｈｙｔａ,ＳｅｐｔｏｒｉａＩＴＳ区长度分别为６３０,５６０,５５０,５４０ｂｐ;酶切图谱属间差别明显,属内种间基本一致,暗示传统的分种可能过细,某些种的成立还有商榷的余地,ＰＣＲ－ＲＦＬＰ对确定疑难种属的地位有重要的意义。３属８种１６个菌株的ＲＡＰＤ结果表明,种内图谱相同或图谱类型相同,而种间明显不同,揭示出过去所划分的种的确存在本质的差别,但是否达到种级的差别还有待于进一步研究     

从ITS序列探讨青藏高原特有植物华福花属的亲缘关系

对青藏高原特有的濒危植物华福花（Ｓｉｎａｄｏｘａ ｃｏｒｙｄａｌｉｆｏｌｉａ Ｃ．Ｙ．Ｗｕ，Ｚ．Ｌ．Ｗｕ ｅｔ Ｒ．Ｆ．Ｈｕａｎｇ）的核糖体ＤＮＡ中的内转录间隔区（ＩＴＳ０序列及５．８ＳｒＲＮＡ基因的序列进行了测定。同川续断目和五加目有关类比较及分支分析表明华福花属与五福花属（Ａｄｏｘａ Ｌ．）近缘，不支持它可能与五加目或败酱科有亲缘关系的假设。尽管形态上它与五福花属分化十分明显，便ＩＴＳ碱基分     

水稻重复序列RRD3在转基因植物中的启动子功能

来源于水稻（Ｏｒｙｚａ ｓａｔｉｖａ Ｌ．）的一个８２０ｂｐ多拷贝重复序列ＲＲＤ３,含有植物启动子ＴＡＴＡ－ｂｏｘ、ＣＡＡＴ－ｂｏｘ等特征保守基元。用ＲＲＤ３取代Ｔｉ载体ｐＢ１１２１中的ＣａｍＶ ３５Ｓ启动子,通过植物转化鉴定ＲＲＤ３的启动子功能。组织化学分析表明,根癌土壤杆菌（Ａｇｒｏｂａｃｔｅｒｉｕｍ ｔｕｍｅｆａｃｉｅｎｓ（Ｓｍｉｔｈ ｅｔ Ｔｏｗｎｓｅｎｄ） Ｃｏｎｎ）ＬＢＡ４４０４转化后     

Sequence Analysis of the ITS Region of Nuclear Ribosomal DNA (nrDNA) in Chinese Amaranthus and Its Systematic Utility

The sequences of the ITS region (including ITS-1, 5.8S rDNA and ITS-2) of nuclear ribosomal DNA from 15 species of the genus Amaranthus L. and outgroup Celosia cristata L. were determined. The result shows that the size of the ITS region of Amaranthus is from 629 to 632 bp, and the length variation is only found in ITS-1 (250-253 bp). On the basis of phylogenetic analysis of nucleotide sequences, the Amaranthus species in China may be devided into three sections, viz., sect. Spinosus, sect. Amaranthus and sect. Paucestamen; the cultivated species, viz., Amaranthus paniculatus L., A. cruentus L., A. caudatus L. and A. hypochondriacus L. can be treated as the subsp. of A. hybridus L.; A. taishanensis F.Z.Li and A. tenuifolius Willd are both closely related to species in the sect. Paucestamen. The study also indicated that the number of stamens has more phylogenetic information than other characters, such as the number of sepals and dehiscence/indehiscence of fruits in Amaranthus.     

用ITS序列研究杨属各组之间的系统发育关系

杨树是重要的工业用材树种。我国杨树遗传资源丰富 ,分布范围广泛 ,不少种为我国特有。开展杨属系统发育和分子进化研究 ,对丰富的杨树遗传资源保存和利用有着重大意义。杨属 (Ｐｏｐｕｌｕｓ)全世界约 10 0余种 ,属下通常分 5个组[1] 。胡志昂等[2 ] 对杨属不同组间的过氧化物酶同工酶进行了研究 ;李宽钰等[3] 利用ＲＡＰＤ标记技术对白杨组、青杨组、黑杨组 2 0个种作了遗传分析。但是在杨属系统分类上还存在着许多混乱 ,同物异名、同名异物现象相当普遍。本文以杨属 5个派主要代表种为材料 ,用ＰＣＲ产物直接测序法测定杨树ＩＴＳ序列 ,…     

帘蛤科贝类rDNA内转录间隔区序列的研究

根据18SrDNA、5．8SrDNA和28SrDNA保守序列设计引物,应用聚合酶链式反应（PCR）扩增了文蛤（Meretrix meretrix L．）、青蛤（Cyclina sinensis G）、硬壳蛤（Mercenaria mercenaria L．）和江户布目蛤（Protothaca jedoensis L．）4种帘蛤科贝类的第一内转录间隔区（ITS1）和第二内转录间隔区（ITS2）序列,并进行了测序。结果表明,文蛤、青蛤、硬壳蛤和江户布目蛤的ITS1扩增产物大小分别为978bp、663bp、757bp和942bp,GC含量分别为61．55％、60．78％、62．48％和64．86％～64．97％,其中ITS1序列长度分别为900bp、585bp、679bp和864bp,是迄今已报道双壳贝类中变化范围最大的,GC含量分别为61．67％、61．03％、63．03％和65．51％～65．62％,江户布目蛤种内ITS1序列有个体差异;ITS2扩增产物大小分别为644bp、618～620bp、593bp和513～514bp,GC含量分别为61．18％、61．29％～61．81％、62．73％和61．48％61．60％,其中ITS2序列长度分别为412bp、386～388bp、361bp和281～282bp,GC含量分别为65．29％、65．21％～66．06％、67．87％和67．38％～67．62％,青蛤和江户布目蛤种内ITS2序列有个体差异。4种蛤ITS1和ITS2序列种间差异很大,有明显的长度多态性,ITS2种间序列相似度73．0％～89．1％,与ITS1的种间序列相似度48．7％～81．5％相比略高。此外,在4种蛤ITS1和ITS2序列中各发现2个与rRNA加工有关的保守区。通过对ITS1和ITS2序列的组装获得了4种蛤5．8SrRNA基因完整序列,序列长度都是157bp,GC含量57．96％～58．60％,4种蛤5．8SrRNA基因相对保守,种间序列差异度0-6．0％,共有10个变异位点,其中转换4处,颠换6处,硬壳蛤和江户布目蛤5．8SrRNA基因序列完全相同。以ITS2序列（包含5．8SrRNA和28SrRNA基因部分序列）为标记,调用北极蛤科的Arctica islandica相应序列数据作外群,构建了帘蛤科贝类的系统发育树,其拓扑结构显示江户布目蛤与硬壳蛤亲缘关系最近,青蛤与其他3物种的亲缘关系最远。     

Study on Sequences of Ribosomal DNA Internal Transcribed Spacers of Clams Belonging to the Veneridae Family (Mollusca: Bivalvia)

The first and second internal transcribed spacer (ITS1 and ITS2) regions of the ribosomal DNA from four species, Meretrix meretrix L., Cyclina sinensis G., Mercenaria mercenaria L., and Protothaca jedoensis L., belonging to the family Veneridae were amplified by PCR and sequenced. The size of the ITS1 PCR amplification product ranged from 663 bp to 978 bp, with GC contents ranging from 60.78% to 64.97%. The size of the ITS1 sequence ranged from 585 bp to 900 bp, which is the largest range reported thus far in bivalve species, with GC contents ranging from 61.03% to 65.62%. The size of the ITS2 PCR amplification product ranged from 513 bp to 644 bp, with GC contents ranging from 61.29% to 62.73%. The size of the ITS2 sequence ranged from 281 bp to 412 bp, with GC contents ranging from 65.21% to 67.87%. Extensive sequence variation and obvious length polymorphisms were noted for both regions in these species, and sequence similarity of ITS2 was higher than that of ITS1 across species. The complete sequences of 5.8S ribosomal RNA gene were obtained by assembling ITS1 and ITS2 sequences, and the sequence length in all species was 157 bp. The phylogenetic tree of Veneridae clams was reconstructed using ITS2-containing partial sequences of both 5.8S and 28S ribosomal DNA as markers and the corresponding sequence information in Arctica islandica as the outgroup. Tree topologies indicated that P. jedoensis shared a close relationship with M. mercenaria and C. sinensis, a distant relationship with other species.     

Evolution of Internal Transcribed Spacer Region of Nuclear Ribosomal DNA in Allopolyploids of Aegilops

Hybridization with subsequent polyploidy is a prominent process in evolution of higher plants, but few data address the evolution of homeologous sequences after polyploidy. The internal transcribed spacer (ITS) of nuclear ribosomal DNA (nrDNA) from eleven allopolyploid species in Aegilops was investigated by PCR amplification and direct sequencing. The sequences obtained were used to study the evolution of ITS region in allopolyploid species. The length of ITS region varied from 599 to 606 bp and the number of variable sites was 93, i.e. 51 and 42 for ITS1 and ITS2 re spectively. Some polymorphic sites were observed in polyploid species, and this indicated that the ancestral sequences had not been homogenized completely by concerted evolution. Distance matrix analysis of diploid and polyploid species by neighbor-joining method, using Triticum monococcum as outgroup, resulted in well-resolved neighbor-joining tree indicating that the ITS regions of UUMM and UUSS genome ( sect. Vertebrata) were homogenizing toward those of UU ancestal genome. This result is in agreement with the results of ctyogenetics of Aegilops. On the other hand, the neighbor joining tree including the D-genome group species (sect. Cylindropyrum and sect. Polyeides ) com prised three clades (CC-DDCC, UU-DDMM-DDMMSS-DDMMUU and MM-DDMvMv), which sug gested that concerted evolution was homogenizing the ITS region of the polyploid derivatives to either of their ancestors.     

A phylogenetic study ofPolygonum sect.Tovara (Polygonaceae) based on ITS sequences of nuclear ribosomal DNA

Polygonum sect.Tovara comprises three morphologically very similar species;P. virginianum,P. filiforme, andP. neofiliforme. Sequences of internal transcribed spacers (ITSs) of nuclear ribosomal DNA of these were determined to examine phylogenetic relationships and the levels of differentiation among them. The size of ITS 1 was 241 bp inP. filiforme andP. neofiliforme, and 242 bp inP. virginianum. The size of ITS 2 was 243 bp, and that of the 5.8S rRNA coding region was 163 bp. The ITS sequences clearly separate North AmericanP. virginianum from the eastern Asian species. Nucleotide divergence between them ranges from 3.3% to 3.8% for ITS 1 and from 9.3% to 10.7% for ITS 2. The molecular data also revealed that two eastern Asian species are closely related but should be treated as distinct species.     

基于ITS序列探讨珍珠菜属过路黄组的系统关系

测定了紫脉过路黄、临时救和过路黄的nrDNA ITS序列,并分析了珍珠菜属过路黄组17个物种的遗传距离及亲缘关系。结果表明,过路黄组植物的ITS序列长度在620～628间,一致性高达90.59%,种间遗传距离为0.002～0.199。系统发育树表明:(1)紫脉过路黄、临时救和小茄亲缘关系较近;(2)大叶过路黄、落地梅和过路黄亲缘关系较近;(3)山萝过路黄、贯叶过路黄、管茎过路黄、叶头过路黄、峨眉过路黄及三角叶过路黄亲缘关系较近。ITS序列分析结果为组内植物的鉴定、分类及系统进化提供了新的参考。     

Circumscription and phylogeny of Saxifraga sect. Ciliatae: Evidence from nrDNA ITS sequences

The largest section of the genus Saxifraga (Saxifragaceae), sect. Ciliatae, consists of 175 morphologically diverse species. This section is mainly distributed in the Qinghai-Tibetan Plateau and adjacent regions of southwest China and more than 80% of the total number of species are endemic to this region. It remains unknown whether this section is monophyletic and up to now no study has been conducted on the infra-sectional phylogeny. In this study, ITS sequences of the nuclear ribosomal DNA were firstly determined for 33 species mainly from this section and related sections. We further downloaded the corresponding sequences of the same DNA region for the other 22 species of Saxifraga and Mitella from GenBank. All sequences were together used to construct the phy-logenetic trees. The main implications of the phylogenetic analyses include: (1) sect. Ciliatae, as traditionally defined, constitutes as a monophyletic clade and its sister group is a well supported clade that includes species from 8 sections such as sect. Porphyrion, sect. Saxifraga and sect. Mesogyne; (2) three morphological subsections, i.e., subsect. Gemmiparae, subsect. Hirculoideae and subsect. Rosulares were tentatively recovered despite the relatively low statistic bootstrap support for the last one; however, subsect. Flagellares and subsect. Hemi-sphaericae were not recognized as separate entities, and nested within subsect. Gemmiparae; (3) subsect. Hircu-loideae and subsect. Rosulares clustered together as sister subclades while subsect. Gemmiparae diverged early. In addition, our results suggest that the paired variation of ITS sequences in sect. Ciliatae is relatively low between the sampled species in spite of their diverse morphology. It is suggested that such a scenario may mirror rapid speciation in this section that probably trigged by the uplifts of the Qinghai-Tibetan Plateau and the extensive selection pressure under the alpine environments.     

报春花属藏报春组、毛茛叶报春组的界定和系统发育关系:核糖体DNA ITS序列证据

报春花属(Primula L.)藏报春组(sect. Auganthus Pax ex Balf. f.)和毛茛叶报春组(sect. Ranunculoides Chen et C. M. Hu)的界定一直是没有解决的问题.应用核糖体DNA ITS序列数据探讨其系统发育关系.取样包括藏报春组和毛茛叶报春组的全部5个种以及其他一些相关组的代表种.ITS系统树表明,陕西羽叶报春(P.filchnerae Knuth)应与藏报春(P. sinensis Sabine ex Lindl.)和野藏报春(P. rupestris Balf. f. et Farrer)一起置于藏报春组;毛茛叶报春组只含两种:毛茛叶报春(P. cicutariifolia Pax)和安徽羽叶报春(P. merrilliana Schltr.).这两组并不具密切亲缘关系,它们在报春花属中与其他组的关系还需进一步研究.研究也表明ITS序列可以为报春花属的系统发育重建提供大量可靠资料.     

用ITS序列确定小麦B基因组的可能供体间的关系

对小麦B基因组的可能供体山羊草属Aegilops sect．Sitopsis的5个种的核糖体DNA的内部转 录区(ITS)进行了PCR扩增和克隆,井测定ITSl和ITS2的序列,用ITSl+ITS2的序列重建了 Aegilops sect．Sitopsis中5个种的系统发育关系。结果表明,斯卑尔脱山羊草Ae．speltoides是sect． Sitopsis中特殊的一个种,它与该组其余4种间的平均遗传距离是后者彼此间平均遗传距离的3倍,Ae． speltoides与同组其余4个种的分离要比后者相互间的分离早得多;在拟斯卑尔脱组Sect．Sitopsis的5 个种中,长柱山羊草Ae．longissima与沙融山羊草Ae．sharonensis的关系最近。ITS序列可以进一步用来作为确定B基因组起源的分子标记。