紫、红黄肉甘薯种质遗传多样性的ISSR分析

采用ISSR分子标记,分析了21份紫肉、28份红黄肉甘薯种质遗传多样性。结果表明：17对引物共扩增出154条谱带,其中多态性谱带138条,占89.6%,平均每个引物扩增出8.12条多态性谱带,表现出丰富的多态性。聚类分析和主成分分析将49份甘薯种质聚为4大类,类型间遗传差异较大,将红黄薯单独聚为1类,说明紫薯和红黄薯分别具有明显不同的来源和系统演化关系。种质间的遗传相似系数变幅为0.58~0.93,其中,0.61~0.70之间的种质占51.4%,0.71~0.80之间的占44.0%。而邻近地域育种单位或同一育种单位的品种亲缘关系较近。文章对如何在育种中利用这些优异种质进行了讨论。     

A molecular tool for genetic surveys in the red coral (Corallium rubrum): An Inter-Simple Sequence Repeats (ISSRs) perspective

Corallium rubrum is a colonial cnidarian, distributed in Mediterranean and Atlantic rocky bottoms. The species has been largely exploited for use in jewellery since 17th century, raising increasing concerns for the sustainability of the present rate of harvesting. The aim of this study was to standardise a simple and not expensive molecular technique to perform genetic analysis on populations of C. rubrum, in order to provide a useful tool for future surveys. Twelve samples collected from one shallow-water and two deep sites located at the Capo Caccia–Isola Piana MPA (North-western Sardinia, Italy) were surveyed by means of 10 ISSR (Inter-Simple Sequence Repeat) primers. Our results confirmed the ease of use and the reliability of ISSR markers, and their usefulness in detecting genetic variability even among individuals.     

38份晾晒烟种质资源遗传关系的SSR分析

利用SSR标记技术对38份晾晒烟种质资源的遗传关系进行了分析。从自己开发的近3000对烟草SSR引物中随机选出30对引物,在38份供试材料中共检测出173个等位基因,每对SSR引物可检测的等位基因数为2~11个,平均为5.77个。38份材料间遗传相似系数（GS）的变化范围为0.165-0.928,平均GS 为0.546。表明38份晾晒烟的遗传多样性丰富,遗传差异较大,亲缘关系较远。聚类分析表明,在L1（GS-0.165）处可将38个品种分为2大类,即晾晒烟类群和美国从烟草起源地收集的烟草（TI：Tobacco Instruction）类群;晾晒烟类群又可进一步分为4组,其聚类结果与所期望的结果基本一致。表明SSR是一种有效、稳定和可靠的分子标记,能较好地从分子水平上揭示烟草(尤其是晾晒烟)种质资源的遗传背景和亲缘关系。     

Genetic structure of a wide-spectrum chicken gene pool

The genetic structure of 65 chicken populations was studied using 29 simple sequence repeat loci. Six main clusters which corresponded to geographical origins and histories were identified: Brown Egg Layers; predominantly Broilers; native Chinese breeds or breeds with recent Asian origin; predominantly breeds of European derivation; a small cluster containing populations with no common history and populations that had breeding history with White Leghorn. Another group of populations that shared their genome with several clusters was defined as 'Multi-clusters'. Gallus gallus gallus (Multi-clusters), one of the subspecies of the Red Jungle Fowl, which was previously suggested to be one of the ancestors of the domesticated chicken, has almost no shared loci with European and White Egg layer populations. In a further sub-clustering of the populations, discrimination between all the 65 populations was possible, and relationships between each were suggested. The genetic variation between populations was found to account for about 34% of the total genetic variation, 11% of the variation being between clusters and 23% being between populations within clusters. The suggested clusters may assist in future studies of genetic aspects of the chicken gene pool.     

吉林省天然黄檗种群遗传多样性ISSR分析

黄檗(Phellodendron amurense Rupr.)是吉林省长白山林区珍贵用材树种和主要建群树种,由于过度采伐和利用,其资源数量和质量明显下降。依据其资源自然分布现状,选择了10个具有代表性的天然黄檗分布种群,应用ISSR标记技术对其进行了遗传多样性的分析,以期为黄檗种群资源的收集、保存和保护提供依据和支持。研究结果表明:从60条ISSR引物中筛选出扩增99条带,多态性条带数为54条,多态性比率为54.5%。10个种群的多态位点比率分布在18.52%—37.96%范围内,其中珲春种群的多态位点比率最高,为37.96%,吉林省露水河种群的多态位点比率最低为18.52%,种群的平均多态位点百分比为26.02%。利用Shannon指数与Nei指数可较好的估算黄檗种群间的遗传变异,Shannon指数的变化范围在0.1103—0.1949之间,Shannon指数总体平均值为0.1522。Nei指数的变化范围在0.0759—0.1327之间,平均为0.1043。根据Nei法计算黄檗10个种群遗传多样性是Dst=0.1586,分化指数Gst=0.6183,基因流系数Nm为0.3086,总的遗传变异中有61.83%的变异存在于群体间,群体内的变异只占38.17%,种群间存在明显分化。黄檗的10个种群可分为两个大群,即:1松江河、露水河、湾沟、集安、辉南2白石山、汪清、安图、延吉、珲春。根据黄檗的遗传结构提出了保护措施:适度引导营造药用或用材林;开展本地黄檗资源的本底调查并进行资源汇总(包括林班、小班,每株的树龄、树高、胸径、枝下高和冠幅等数据),筛选本地的优势群体进行原地保存;迁地保护策略中要增加样本的数量,白山地区迁地保护的种源应选择松江河、露水河种源,通化地区迁地保护的种源应选择集安种源,而延边地区应选择白石山和汪清种源;人工促进黄檗的天然林更新改造,逐步恢复黄檗种群规模,并且进行人工更新的资源登记。     

The long and the short of gene flow and reproductive isolation: Inter-Simple Sequence Repeat (ISSR) markers support the recognition of two floral forms in Pelargonium reniforme (Geraniaceae)

The current taxonomy of Pelargonium reniforme recognises two subspecies on the basis of habit and vegetative characters, but excludes floral characters. However, populations of P. reniforme in the wild tend to belong to easily discernable floral groups based on floral colour and hypanthium length. The aim of this study was to determine the genetic diversity within and between populations of both subspecies using the Inter-Simple Sequence Repeat (ISSR) method of DNA fingerprinting. Ninety individuals from eight populations were sampled. Both phenetic and Neighbor-Net analyses reveal that populations are genetically discernable, but that there is no genetic evidence to support the recognition of the two currently defined subspecies of P. reniforme. Instead, the analyses resolved all individuals that possess long hypanthia into a single group, and the role of different pollinators in driving reproductive isolation of this long-tubed form is suggested.     

人腺病毒B、D亚种基因组中简单重复序列分析

本文以人腺病毒B亚种31条基因组序列及D亚种39条基因组序列为研究材料,利用ImperfectMicrosatelliteExtractor和DNAMAN软件对这些基因组序列中简单重复序列(SSR)的分布情况进行了系统性分析和比较。分析结果显示:人腺病毒B、D亚种基因组中简单重复序列的平均相对密度是十分接近的,但在不同类型SSR中分布情况又有所不同。D亚种中二型SSR明显高于B亚种,在两亚种一型SSR中(A)n、(T)n都是比较多的,而在两亚种二型SSR中的(CG/GC)n表现出了较高的偏好性。在同亚种多序列比对分析中,D亚种表现出了更高的稳定性。B、D亚种中SSR的这种特异性分布可能与它们的进化机制和致病性有关。     

若尔盖湿地青海沙蜥红原亚种线粒体ND4-tRNAleu基因的序列变异和遗传多样性

研究了青海沙蜥红原亚种(Phrynocephalus vlangalii hongyuanensis)分布于四川若尔盖辖曼(XM)、红原(HY)和甘肃玛曲(MQ)这3个地理单元7个居群的ND4-tRNAleu基因的序列变异和遗传多样性。72个序列比对得到785bp的片断,含变异位点7个,定义了9种单倍型。结果显示7个居群总的核苷酸多样性较低(0.00231±0.00016),单倍型多样性较高(0.806±0.024);就单个居群而言,MQa、MQb、XMb的遗传多样性低,XMc相对较高。各居群间的Kimura双参数遗传距离小(0.001—0.005),最大遗传距离出现在MQa和XMa之间,XM单元各居群之间遗传距离最小。但分子变异分析(AMOVA)显示3个单元间出现了显著差异(P<0.01),遗传变异主要存在于地理单元间,占62.61%。除MQ单元形成一支,单倍型网络图没有显示出单倍型和地理位置的对应关系,XM各居群以及HY混杂在一起。XM单元单倍型的不配对分布(Mismatch distribution)为明显左移的单峰,同时Fu’sFstest得到负值(Fs=-2.21937),这都暗示XM单元可能经历了近期种群扩张,这是其单倍型多样性较高和核苷酸多样性较低的原因。MQ单元遗传多样性低而与其他单元显著分化,推测与黄河在若尔盖玛曲之间贯通有关。近期沼泽的形成对XMb的隔离时间短,使得其遗传多样性低但还不足以形成大的遗传差异。     

High levels of diversity in Fusarium oxysporum from non-cultivated ecosystems in Australia

The Fusarium oxysporum species complex (FOSC) is a ubiquitous ascomycetous group that includes both pathogenic and non-pathogenic strains, the former being responsible for disease in over 100 cultivated plant species. Previous phylogenetic studies have uncovered at least four major clades within the FOSC, with Clade 1 hypothesised as being ancestral. However, the origin of these clades and pathogenic strains is poorly understood. Due to an emphasis on agricultural isolates in previous studies, the underlying diversity of this species complex in non-cultivated soils is largely unknown. To address this imbalance an extensive survey of isolates associated with native vegetation geographically isolated from cultivation throughout the Australian continent was conducted. A multi-gene phylogenetic analysis of the translation elongation factor (EF-1α) and the mitochondrial small subunit (mtSSU) rDNA loci did not recover any novel clades. However, the Australian isolates had high levels of intra-Clade diversity based on EF-1α sequence type (ST) comparison with a global dataset. The ST diversity was not equally distributed across the four clades, with the majority of novel STs recovered from Clade 1. Implications on the origin of the FOSC are discussed.     

AFLP and breeding system studies indicate vicariance origin for scattered populations and enigmatic low fecundity in the Moroccan endemic Hypochaeris angustifolia (Asteraceae), sister taxon to all of the South American Hypochaeris species

We used Amplified Fragment Length Polymorphism markers (AFLP) and breeding system studies to investigate the population structure and reproductive biology of Hypochaeris angustifolia (Asteraceae: Cichorieae). This species is endemic to altiplanos of the Atlas Mountains (Morocco) where it occurs in scattered populations, and it is the sister species to c. 40 species of this genus in South America. PCoA, NJ, and Bayesian clustering, revealed that the populations are very isolated whilst AFLP parameters show that almost all populations have marked genetic divergence. We contend that these features are more in accord with a vicariance origin for the scattered populations of H. angustifolia, rather than establishment by long-distance dispersal. The breeding system studies revealed that H. angustifolia is a self-incompatible species, with low fecundity in natural and in experimental crosses, probably due to a low frequency of compatible phenotypes within and between the populations.     

马鹿四个亚种的起源和遗传分化研究

利用线粒体ＤＮＡ的细胞色ｂ基因片段序列差异来分析探讨马鹿４个亚种的起源和遗传分化问题。结果表明：马鹿各亚种的ＤＮＡ差异都达到了亚种分化水平,并指出天山马鹿东西两个种群可能是两个不同的亚种;马鹿各亚种在５４－７６万年前开始分化,其中天山马鹿最先分化出来;     

Sequence variations at a complex microsatellite locus in rice and its conservation in cereals

In an attempt to study changes associated with microsatellites in rice, the DNAs of cultivated rice, including indica and japonica varieties, and wild rice genotypes were amplified by the polymerase chain reaction with primers flanking the (GATA) _n and (AC) _n repeats at a microsatellite-containing locus OS1E6 (Genebank accession number AFO16647) previously reported from a PstI rice (var. Malkolam) genomic library in pUC18. Eight alleles of varying sizes were obtained which were cloned and sequenced. Sequencing data indicated that the size variations of the different alleles were due to differences in the repeat number as well as to sequence variations in the region flanking the microsatellite motifs. In order to study the presence of this complex microsatellite-containing locus of rice in different cereals, their DNAs were amplified using primers flanking the OS1E6 locus. It was found that this locus was present in the various cereal genotypes analyzed, indicating its conservation across different cereal members. Received: 10 March 2000 / Accepted: 14 April 2000     

缢蛏种群遗传多样性的周年变异分析

利用线粒体COI标记分析了福建省宁德市漳湾镇横屿村滩涂5个月份缢蛏群体(S3,S5,S7,S9,S11)的遗传结构,以期评估不同月份时期种群的遗传多样性差异。基于线粒体COI基因的结果表明,5个缢蛏群体的平均核苷酸差异数位于2.7836-3.6894之间,核苷酸多样性指数位于0.0050-0.0066之间,遗传多样性水平大致表现为S3和S5群体较高于S7和S9群体,明显高于S11群体。AMOVA分析结果显示,群体间遗传变异量占总变异的7.18%,而群体内变异达到了92.82%,说明遗传差异主要来自于群体内部。由此可见,从3月份到11月份,缢蛏群体的遗传多样性水平呈现出下降趋势,尤其是在11月份,差异最为明显。     

Genetic diversity and population structure of food-borne Staphylococcus carnosus strains

The species Staphylococcus carnosus is a non-pathogenic representative of the coagulase negative staphylococci. Specific strains are applied as meat starter cultures. The species consists of two subspecies, S. carnosus ssp. carnosus and S. carnosus ssp. utilis. In order to place S. carnosus strains, characterized in former studies, in a genetic background that allows a typing of candidates for starter cultures, a Multilocus Sequence Typing (MLST) scheme was developed. Internal fragments of the genes tpiA, xprT, dat, gmk, glpK, narG, cstA, encoding triosephosphate isomerase, xanthine phosphoribosyltransferase, d-amino acid aminotransferase, guanylate kinase, glycerol kinase, the α-chain of the respiratory nitrate reductase, and a carbon starvation protein where chosen. Genes were selected based on their equal distribution in the genome, taxonomic value in subspecies differentiation and metabolic function. This MLST was applied to 44 S. carnosus strains, most of them previously analyzed for their suitability as starter cultures.The number of alleles varied between zero (tpiA) and five (cstA) and allowed the definition of nine sequence types (ST). ST1 was most abundant (18 strains), followed by ST2 (8) and ST4 (6). The nine STs confirmed a close relationship of all strains despite their isolation source and year, but lacked correlation with physiological activities relevant for starter cultures. The low amount of STs in the strain set lets us suggest that recombination between strains is rare. Thus, it is hypothesized that evolutionary events seem to be due to single point mutations rather than intrachromosomal recombination, and that the species possesses a clonal population structure.     

A novel program to design siRNAs simultaneously effective to highly variable virus genomes

A major concern of antiviral therapy using small interfering RNAs (siRNAs) targeting RNA viral genome is high sequence diversity and mutation rate due to genetic instability. To overcome this problem, it is indispensable to design siRNAs targeting highly conserved regions. We thus designed CAPSID (Convenient Application Program for siRNA Design), a novel bioinformatics program to identify siRNAs targeting highly conserved regions within RNA viral genomes. From a set of input RNAs of diverse sequences, CAPSID rapidly searches conserved patterns and suggests highly potent siRNA candidates in a hierarchical manner. To validate the usefulness of this novel program, we investigated the antiviral potency of universal siRNA for various Human enterovirus B (HEB) serotypes. Assessment of antiviral efficacy using Hela cells, clearly demonstrates that HEB-specific siRNAs exhibit protective effects against all HEBs examined. These findings strongly indicate that CAPSID can be applied to select universal antiviral siRNAs against highly divergent viral genomes.     

黄山短尾猴mtDNA控制区序列变异及种群的遗传多样性

短尾猴属灵长目(Primates)猴科(Cercopithecidae)猕猴属(Macaca),是我国特有的国家二级保护动物。为了更有效地保护其野生种群,本文研究了黄山短尾猴种群内的遗传多样性,并对黄山短尾猴与四川短尾猴种群间的遗传差异进行了分析。共测定了黄山短尾猴7个群体中的30个样本的mtDNA控制区5′端493bp的序列,只发现了7个变异位点,定义了3种单倍型,单倍型序列之间缺乏变异,种群中的核苷酸多样性很低(0.006);3种单倍型相应地将黄山种群分为了3个亚群,不同亚群之间呈现出一定的片断化分布,从分子水平上初步揭示了短尾猴黄山种群的遗传多样性。与四川短尾猴的相应序列比较,黄山短尾猴控制区序列存在很大差异,共有59个变异位点,而且存在大片段的碱基插入/缺失,有78%的遗传变异发生在两个种群之间,两个种群间的核苷酸歧异度已达8.21%。进一步分析表明,黄山短尾猴与四川短尾猴之间存在着极显著的遗传分化(FST=0.399,P<0.001),基于最大似然法和邻接法构建的系统发生树均将两者聚为不同的类群,支持将它们归入各自的管理单元。     

Coprophilous fungi from Morocco

Summary

Fifty-seven species of coprophilous fungi are recorded from 14 dung samples collected from the Souss Valley area of southern Morocco that were incubated in moist chambers. Several new records for Morocco are reported. Evidence for reduced diversity due to the severely degraded nature of the habitats in which the samples were collected is discussed.     

Natural hybridization within seed sources of shortleaf pine (Pinus echinata Mill.) and loblolly pine (Pinus taeda L.)

Shortleaf and loblolly pine trees (n = 93 and 102, respectively) from 22 seed sources of the Southwide Southern Pine Seed Source Study plantings or equivalent origin were evaluated for amplified fragment length polymorphism (AFLP) variation. These sampled trees represent shortleaf pine and loblolly pine, as they existed across their native geographic ranges before intensive forest management. Using 17 primer pairs, a total of 96 AFLPs between shortleaf pine and loblolly pine were produced and scored on the sample trees and two control-pollinated F1 interspecies hybrids and their parents. In addition, the well known isocitrate dehydrogenase (IDH) isozyme marker was scored for all trees. IDH detected two putative hybrids among the loblolly pine samples and two among the shortleaf pine samples, while either 13 or 12 putative hybrids were detected using all AFLP markers and IDH and either NewHybrids or Structure software, respectively. Results of this study show that later generation hybrids can be reliably identified using AFLP markers and confirmed that IDH is not a definitive marker for detecting hybrids; that is, at least in some seed sources, the alternative species’ IDH allele resides in the source species. Based on all the markers, hybridization frequency varied geographically, ranging from 30% in an Arkansas seed source to 0% in several other seed sources. The hybridization level was higher in populations west of the Mississippi River than in populations east of the river; the shortleaf pine hybridization rates were 16.3% and 2.4% and the loblolly pine rates were 4.5% and 3.3%, west and east of the river, respectively. The results suggest that hybridization between these two species is significant but varies by seed source and species, and the potential for the unintended creation of hybrids should be considered in forest management decisions regarding both natural and artificial regeneration.