杨属(Populus L.)种质资源遗传学评价研究进展

杨属(Populus L.)种质资源极其丰富, 为了有效保存、合理利用杨属种质资源, 国内外开展了大量的种质资源遗传学评价研究。该文在介绍杨树系统分类的基础上, 概述了白杨派、青杨派和黑杨派等在生物学特性、抗性(耐盐、抗旱、抗冻及抗病虫)、适应性及DNA遗传多态性等方面的遗传学评价研究进展, 重点讨论了杨树种质资源评价研究中存在的问题和不足, 并对其研究前景进行了展望。     

杨属（PopulusL.）种质资源遗传学评价研究进展

杨属（PopulusL.）种质资源极其丰富,为了有效保存、合理利用杨属种质资源,国内外开展了大量的种质资源遗传学评价研究。该文在介绍杨树系统分类的基础上,概述了白杨派、青杨派和黑杨派等在生物学特性、抗性（耐盐、抗旱、抗冻及抗病虫）、适应性及DNA遗传多态性等方面的遗传学评价研究进展,重点讨论了杨树种质资源评价研究中存在的问题和不足,并对其研究前景进行了展望。     

应用SRAP标记分析新疆地区主要杨属树种的遗传多样性

用30对SRAP引物对15个采集于新疆不同地区的杨属树种进行了遗传多样性分析,共扩增出881条具多态性的条带,平均多态性信息量为0．958,平均多态性比率为96．8％．通过聚类分析将15个杨树材料分为4个群．这一聚类结果与传统的分类基本上一致。这表明SRAP标记适用于研究杨属树种遗传多样性。     

西南地区古杨树遗传多样性的SSR分析

采用SSR分子标记技术,对采集于中国西南地区的7种15个群体共226株杨树古树的遗传多样性进行分析,以揭示杨树古树的遗传变异水平及其遗传关系,为该区域杨树古树资源的合理保护与利用提供依据。结果显示:(1)7对SSR引物组合共扩增出80个多态性位点;以有效等位基因数为标准,7种杨树之间的遗传多样性水平依次为:西南杨昌都杨康定杨藏川杨乡城杨川杨德钦杨;杨树种内群体间的遗传多样性依次表现为:乡城杨的稻城群体乡城群体雅江群体,西南杨的理塘群体乡城群体稻城群体,德钦杨的香格里拉群体德钦群体,昌都杨的芒康群体贡觉群体昌都群体,藏川杨的德钦群体芒康群体。(2)乡城杨和西南杨群体内近交系数(F_(is))分别为-0.024 6和-0.253 5,藏川杨、德钦杨和昌都杨的Fis分别为0.205 4、0.240 1和0.029 2;乡城杨、西南杨、藏川杨、德钦杨和昌都杨群体间的遗传分化系数(F_(st))分别为0.156 1、0.253 5、0.128 8、0.182 0和0.177 3;除西南杨群体间基因流N_m小于1外,乡城杨、藏川杨、德钦杨和昌都杨群体间的基因流N_m均大于1。(3)7种杨树古树的遗传相似系数介于0.089 0~0.691 0,平均为0.402 0;UPGMA及Bayesian聚类结果均显示,7个树种可以分为3个大组,即川杨与康定杨聚为一组,乡城杨与西南杨聚为一组,藏川杨、德钦杨和昌都杨聚为第3组;5种杨树古树的群体UPGMA聚类结果显示,除西南杨的理塘群体与乡城杨聚为一组外,各杨树古树种与群体间的聚类关系较为紧密。研究表明,西南地区杨树古树具有较为丰富的遗传多样性,且其遗传变异主要均存在于群体内不同个体之间,任何个体的丧失均会导致杨树古树遗传多样性的降低或丧失,故需要加强对该区域现存古树资源的保护。     

基于cpDNA和rDNA ITS片段分析西南地区青杨派杨树的系统发育与进化关系

西南地区青杨派杨树种质资源丰富,可为杨树遗传改良提供珍贵的基因资源,但树种之间形态学差异细微,该研究以山杨作为外类群,测定了西南地区及其他地区杨属青杨派17个种(杂种)共36份样本的3个叶绿体片段(atpF-atpH、trnL-F和matK)和核糖体ITS片段,并对其进行系统发育分析,以探讨西南地区青杨派树种的系统进化关系。结果表明:(1)在所有样本中,3个叶绿体片段atpF-atpH、trnL-F、matK的长度分别为605~634bp、957~1 010bp、819bp,3个片段拼接后的联合序列包含29个变异位点和15个信息位点;ITS片段对齐后的长度为646bp,变异位点19个,信息位点17个。(2)基于叶绿体联合序列和ITS片段的杨属青杨派树种的平均遗传距离分别为0.001 3和0.003 6。叶绿体联合片段的MP和Bayes系统树树型基本一致,青杨派树种可以划分为2组,第1组由青杨、三脉青杨、大青杨和辽杨构成;第2组中的小叶杨、小青杨、川杨、德钦杨、昌都杨、乡城杨、康定杨、西南杨、滇杨和藏川杨不能有效区分,且均与缘毛杨的遗传关系较近。(3)基于ITS的系统树与叶绿体联合片段构建的系统树差异不大,仅在于第2组中的小青杨与小叶杨、川杨等差异明显,与第1组中的树种紧密聚拢。     

应用微卫星标记分析中国地方鸡种的遗传变异

利用 8个微卫星位点对中国 9个地方鸡种和 1个引进品种进行了遗传检测。计算出了各品种的平均杂合度、平均多态信息含量 (PIC)及品种间的遗传距离 ,并进行了系统聚类。结果表明 :8个微卫星位点上共检测到了5 4个等位基因 ,每个位点上平均为 6 .75个。各位点平均多态信息含量为 0 .5 0 71～ 0 .74 34,均表现出了高度多态性。各群体平均杂合度较高 ,为 0 .5 5 6 4～ 0 .7135 ,说明我国地方鸡种有着较丰富的遗传多样性。地方鸡种间的遗传距离相对较远 ,10个鸡种共分为三大类。研究结果对我国鸡种资源的评估、保存和预测杂种优势具有一定的指导意义     

杨属植物化学成分的研究概况

杨树是我国重要的绿化造林用树。在温带地区,蜂胶的植物来源主要是杨属及其杂交属的芽孢分泌物。有不法商人以杨树芽提取物冒充蜂胶,严重阻碍了蜂胶行业的健康发展。本文对杨属植物中已分离鉴定出的化学成分进行了归类整理,为蜂胶和杨属化学成分及药理活性的研究提供依据,同时,也为杨树在医药保健等方面的综合利用提供理论基础。     

我国野生稻酒昔酶同工酶的初步研究

我国野生稻资源丰富。丁颖等认为我国野 生稻有三个种,即：普通野生稻(Oryza sativa L. f. spontanea),药用野生稻(Oryza of ficinalis Wall),沈粒野生稻(Oryza meyeriana Baill^[1,2] 近年来野生稻资源的利用已被国内外的遗传育 种工作者所重视,对于野生稻资源的收集和研 究不仅在理论上,而且在生产实践上都具有重 要意义。本工作利用聚丙烯酸胺凝胶电泳对 1980年收集的我国野生稻的三个种进行醋酶 同工酶分析研究,获得了初步的结果。     

杨树抗冻性的研究现状

杨树是世界上广泛栽培的重要造林树种之一,我国是世界上杨树资源丰富的国家。杨树具有速生丰产、实用性强、无性繁殖能力强,且基因组较小等特点,现已成为研究林木生理和基因工程研究的模式植物。本文概述了国内外有关杨树抗冻性的生理生化性质及其指标、天然杂种的选择以及人工杂交选育种等方面研究现状,并展望了杨树基因工程在杨树抗冻性遗传改良中的应用前景。此外,对今后如何开展杨树抗冻改良育种的研究提出了一些设想。     

杨树抗冻性的研究现状

杨树是世界上广泛栽培的重要造林树种之一,我国是世界上杨树资源丰富的国家,杨树具有速生丰产、实用性强、无性繁殖能力强,且基因组较小等特点,现已成为研究林木生理和基因工程研究的模式植物。本文概述了国内外有关杨树抗冻性的生理生化性质及其指标、天然杂种的选择以及人工杂交选育种等方面研究现状,并展望了杨树基因工程在杨树抗冻性遗传改良中的应用前景,此外,对信后如何开展杨树抗冻改良育种的研究提出了一些设想。     

用ITS序列研究杨属各组之间的系统发育关系

ITS sequences of 15 representative species of five sections inthe genus Populus L. were determined. By using direct sequencing of PCR product, it was found that the fragments of internal transcribed spacers (ITS) are about 594 bp in length. The length of ITS-1 and ITS-2 is about 220 bp and 210 bp, respectively, while that of 5.8s is 164 bp. Its G+C content is about 69.0%. The number of phylogenetically informative loci is higher in ITS-2 than in ITS-1. Transversion and transition are two main factors that drive the ITS evolution, and more insertions and deletions occurred in ITS-2. Taking Salix matsudana Koidz. and Salix suchowensis Cheng as outgroups, phylogenetic analysis of ITS sequences using PAUP 4.0 software indicated that Populus is monophyletic group and can be divided into two main clades. One is the section Leuce, and the other is the remaining sections.     

Phylogenetic Relationship ofPopulusSections by ITS Sequence Analysis

ITS sequences of 15 representative species of five sections in the genus Populus L. were determined. By using direct sequencing of PCR product, it was found that the fragments of internal transcribed spacers (ITS) are about 594 bp in length. The length of ITS-1 and ITS-2 is about 220 bp and 210 bp, respectively, while that of 5.8s is 164 bp. Its G+C content is about 69.0%. The number of phylogenetically informative loci is higher in ITS-2 than in ITS-1. Transversion and transition are two main factors that drive the ITS evolution, and more insertions and deletions occurred in ITS-2. Taking Salix matsudana Koidz. and Salix suchowensis Cheng as outgroups, phylogenetic analysis of ITS sequences using PAUP 4.0 software indicated that Populus is monophyletic group and can be divided into two main clades. One is the section Leuce , and the other is the remaining sections.     

中国苋属nrDNA的ITS序列分析及其系统学意义

运用ＰＣＲ直接测序法,对苋属（Ａｍａｒａｎｒｈｕｓ Ｌ．）１５个种及外类群鸡冠花（Ｃｅｌｏｓｉａ ｃｒｉｓｔａｔａ Ｌ．）ｎｒＤＮＡ的ＩＴＳ区（包括ＩＴＳ－１,５．８５ｒＤＮＡ和ＩＴＳ－２）进行序列测定。结果表明苋属植物的ＩＴＳ序列总长度为６２９－６３２ｂｐ,长度变异仅发生在ＩＴＳ－１区（２５０－２５３ｂｐ）。采用ＰＡＵＰ软件进行系统发育分析表明：分布于中国的苋属植物可分为３组,即刺苋组（ｓｅｃｇ     

From Genus to Phylum: Large-Subunit and Internal Transcribed Spacer rRNA Operon Regions Show Similar Classification Accuracies Influenced by Database Composition

We compared the classification accuracy of two sections of the fungal internal transcribed spacer (ITS) region, individually and combined, and the 5′ section (about 600 bp) of the large-subunit rRNA (LSU), using a naive Bayesian classifier and BLASTN. A hand-curated ITS-LSU training set of 1,091 sequences and a larger training set of 8,967 ITS region sequences were used. Of the factors evaluated, database composition and quality had the largest effect on classification accuracy, followed by fragment size and use of a bootstrap cutoff to improve classification confidence. The naive Bayesian classifier and BLASTN gave similar results at higher taxonomic levels, but the classifier was faster and more accurate at the genus level when a bootstrap cutoff was used. All of the ITS and LSU sections performed well (>97.7% accuracy) at higher taxonomic ranks from kingdom to family, and differences between them were small at the genus level (within 0.66 to 1.23%). When full-length sequence sections were used, the LSU outperformed the ITS1 and ITS2 fragments at the genus level, but the ITS1 and ITS2 showed higher accuracy when smaller fragment sizes of the same length and a 50% bootstrap cutoff were used. In a comparison using the larger ITS training set, ITS1 and ITS2 had very similar accuracy classification for fragments between 100 and 200 bp. Collectively, the results show that any of the ITS or LSU sections we tested provided comparable classification accuracy to the genus level and underscore the need for larger and more diverse classification training sets.     

大白口蘑分离菌株的DNA鉴定

以松口蘑Tricholoma matsutake子实体为外类群,对大白口蘑T. giganteum 野生子实体及其组织分离菌丝进行ITS序列测序,通过DNAStar软件进行比较分析。结果表明大白口蘑ITS序列长度为589bp,松口蘑ITS序列长度为601bp,ITS1和ITS2呈现不同程度的种间多态性;ITS序列测定证实了大白口蘑野生子实体及其组织分离菌丝的同质性,并且ITS区序列在大白口蘑种内不同菌株间的变异程度很小,表明使用通用引物ITS4和ITS5,通过PCR扩增测序即可用于大白口蘑的种质鉴定。     

Length Variation of the Nuclear Ribosomal DNA Internal Transcribed Spacer in the Genus Abies,with Reference to Its Systematic Utility in Pinaceae

The internal transcribed spacer (1TS) region (1TS1, ITS2 and 5.8S rDNA) of the nuclear ribosomal DNA (nrDNA) was amplified via PCR in 28 taxa of Abies Mill. The amplified fragments showed length polymorphism among species, with species from Central America and two species from North America having a length of approximately 2 500 base pairs (bp) and the remaining taxa having a length of approximately 1 700 bp based on 100 bp and 1 kb ladder standard markers. The complete sequencing of ITS of Abies bracteata showed that the shorter type is 1 697 bp (1TS1 is 1 296 bp, 5.8S + 1TS2 is 401 bp). For the longer one, the partial rrs1 and complete 5.8S + ITS2 sequencing revealed that thelength of 5.8S + ITS2 is the same as that of the shorter type. The length difference of ITS in Abies is mainly due to the length difference in the ITS1 region, a result similar to the previous findings in other genera of Pinaceae. Variation in ITS length seems well correlated with morphological and geographic characters in Abies, suggesting that the length variation may be a phylogenetically informative character within the genus, long ITS was also found in other genera of Pinaceae in the previous studies. The long length of ITS in the family makes the sequencing of the region and subsequent alignment of sequences among species or genera more difficult than in taxa with short ITS, such as angiosperms. Although the length variation of ITS in the genus Abies is significant, the homogenous of ITS sequence between the longer one and the shorter one is obvious if the insertion in the longer ITS is ignored.     

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。     

Evolution of the secondary structure of the rRNA internal transcribed spacer 2 (ITS2) in hard ticks (Ixodidae, Arthropoda)

ITS2 sequences are used extensively in molecular taxonomy and population genetics of arthropods and other animals yet little is known about the molecular evolution of ITS2. We studied the secondary structure of ITS2 in species from each of the six main lineages of hard ticks (family Ixodidae). The ITS2 of these ticks varied in length from 679 bp in Ixodes scapularis to 1547 bp in Aponomma concolor. Nucleotide content varied also: the ITS2 of ticks from the Prostriata lineage (Ixodes spp.) had 46-49% GC whereas ITS2 sequences of ticks from the Metastriata lineage (all other hard ticks) had 61-62% GC. Despite variation in nucleotide sequence, the secondary structure of the ITS2 of all of these ticks apparently has five domains. Stems 1, 3, 4 and 5 of this secondary structure were obvious in all of the species studied. However, stem 2 was not always obvious despite the fact that it is flanked by highly conserved sequence motifs in the adjacent stems, stems 1 and 3. The ITS2 of hard ticks has apparently evolved mostly by increases and decreases in length of the nucleotide sequences, which caused increases, and decreases in the length of stems of the secondary structure. This is most obvious when stems of the secondary structures of the Prostriata (Ixodes spp.) are compared to those of the Metastriata (all other hard ticks). Increases in the size of the ITS2 may have been caused by replication slippage which generated large repeats, like those seen in Haemaphysalis humerosa and species from the Rhipicepalinae lineage, and the small repeats found in species from the other lineages of ticks.     

基于核rDNA的ITS序列在种子植物系统发育研究中的应用

种子植物核rDNA是高度重复的串联序列,由于同步进化的力量．大多数物种中这些重复单位间已发生纯合或接近纯合。5．8S rDNA把核rDNA的内转录间隔区分为ITS1和ITS2两部分．在被子植物中ITS1的长度为165～298bp,ITS2的长度为177～266bp,而在裸子植物中ITS片段较长。且其长度变化主要由ITS1的长度变异所致。可对这两个片段PCR产物进行直接测序或克隆测序。由于ITS序列变异较快．能够提供较丰富的变异位点和信息位点,已成为被子植物较低分类阶元的系统发育和分类研究中的重要分子标记,为探讨多倍体复合体网状进化关系,异源多倍体的起源提供了重要的系统学信息．但它一般不适合科以上水平的系统学研究。裸子植物中ITS片段较长,重复序列间的纯合程度不同,测序比较困难．因此对探讨裸子植物系统发育和分类受到了一定的限制,但近年来有所发展。     

Molecular evidence for the synonymy of two species of Apatemon Szidat, 1928, A. gracilis (Rudolphi, 1819) and A. annuligerum (von Nordmann, 1832) (Digenea: Strigeidae) parasitic as metacercariae in British fishes

The current study examined rDNA (internal transcribed spacer regions, ITS1 and ITS2) and cytochrome c oxidase subunit 1 (CO1) sequence data of Apatemon annuligerum (originating from two geographical locations) and A. gracilis metacercariae (originating from four natural piscine hosts) to determine the systematic status of these two strigeid digeneans. With the exception of short repeat motifs, the ITS1 regions sequenced demonstrated no intra- or inter-specific sequence variation. ITS2 sequences were 292 bp and CO1 sequences 366 bp in length and identical for both nominal Apatemon species. These sequence data provide strong evidence that the two species are con-specific and that A. annuligerum should be regarded as a junior synonym of A. gracilis.