我国主要花生品种的AFLP分析

对32个来源于中国不同产地的花生品种进行了AFLP指纹图谱及相似性聚类分析．结果表明：所有供试花生品种的遗传相似性为35％，在45％的相似性水平处分为3个群，表明我国的花生品种存在遗传多态性．AFLP指纹图谱分析技术能很好地反映出花生品种之间存在的遗传差异．图2表1参26     

AFLP法分析染料对黄孢原毛平革菌的损伤效应

利用AFLP对DNA碱基序列分辨率高的特性,研究纯培养条件下染料对黄孢原毛平革菌(P. chrysosporium)菌株DNA的损伤效应.在对黄孢原毛平革菌(P. chrysosporium)AFLP最佳引物筛选的基础上,通过分析最佳引物扩增结果中DNA相似性、染料浓度与DNA相似性关系以及构建UPGMA相似性树状图,评价染料对P. chrysosporium DNA损伤效应.结果表明, AFLP法可较好反映染料对DNA损伤效应,在优选的3种引物组合中, E-AAC/M-CAA引物组合AFLP扩增效果最好,共获得35条DNA扩增条带,其中有12条具有特异性,多态性比率34.3%;在进行DNA相似性分析和UPGMA法聚类分析后发现, AFLP技术较好地表达了DNA相似性与染料浓度的正相关关系,并且显示P. chrysosporium对染料损伤效应敏感,染料浓度不宜高于50mg/L,当染料浓度达到300mg/L即可对P. chrysosporium产生较大损伤作用,使之基本丧失原有的处理功能.     

长柄石杉居群遗传多样性和遗传结构的AFLP分析

石杉科植物因所含石杉碱甲(Huperzine A)对中老年痴呆等具有良好疗效,近年来倍受关注.利用AFLP分子标记对武夷山脉广布种长柄石杉[Huperzia serrata(Thunb.ex Murray)Trev.var.longipetiolata(Spring)H.M.Chang]7个居群112株个体进行遗传多样性和居群遗传结构分析.选用多态性高、分辨力强的8对选择性扩增引物组合共获得675个位点,其中多态位点比例为69.38%.居群内观测等位基因数(Na)为1.633,有效等位基因数(Ne)1.493;Nei’s基因多样性指数(He)与Shannon多态性信息指数(I)的平均值分别为0.272和0.392,多样性最高为地处武夷山脉中段的泰宁居群和建宁居群,最低为山脉北段的光泽居群.居群总基因多样性(Ht)为0.327 3,居群内基因多样性(Hs)为0.272 2,居群间的遗传分化系数(Gst)为0.168 1,表明居群内变异是长柄石杉遗传多样性的主要来源.由Gst估计,武夷山脉长柄石杉自然居群的基因流(Nm)为2.474 0.邻接树分析表明居群间遗传亲缘关系与地理位置相关.武夷山脉长柄石杉较高的遗传多样性和基因流水平表明其仍然具有相当的适应(生存)能力和进化潜力,这可能与其生物学(异交水平)、生态学特性及武夷山脉相对良好的生境条件有关.     

松毛虫遗传多样性研究中AFLP反应体系的建立

以油松毛虫为材料,对T4 DNA连接酶不同用量、预扩增中的Mg~(2+)浓度、dNTP浓度、引物浓度以及选择性扩增巾的预扩增产物稀释倍数、Mg2~(2+) 浓度、dNTP浓度、引物浓度和Taq酶浓度进行了比较分析.最终建立了适合松毛虫的AFLP反应体系.用优化的AFLP反应体系,以油松毛虫、赤松毛虫和马尾松毛虫为材料筛选引物,从81对引物组合中筛选出10对多态性高的引物组合.图9参17     

Genetic analysis of parentage within experimental populations of a male dimorphic beetle,<Emphasis Type="Italic"> Onthophagus taurus</Emphasis>, using amplified fragment length polymorphism

We used the multilocus DNA fingerprinting technique, amplified fragment length polymorphism, to examine parentage of 902 offspring from eight experimental populations of the dung beetle, Onthophagus taurus. The males of this species exhibit a suit of morphological and behavioural traits that characterise alternative mating tactics. Hornless minor males sneak copulations with females that are guarded by horned major males. Our aims were to provide a prospective assessment of the potential role of frequency dependence in the maintenance of alternative mating tactics, to assess the levels of polyandry, and to determine the patterns of sperm usage by multiply mated females. The average proportion of offspring sired by major and minor males did not co-vary with the relative frequency of each morph present in experimental populations. However, there was some indication that the effective mating frequency (number of females producing offspring sired by a given male) of major and minor males may exhibit frequency dependence. Mating success of both male types declined with increasing numbers of major males. Paternity was positively associated with effective mating frequency. Females produced offspring sired from between one and eight males and, on average, paternity was distributed equally amongst a females mates, regardless of the number of males mated. Differences in fertilisation success among males were not associated with alternative male phenotypes. Neither did a males fertilisation success depend on his genetic dissimilarity with the female. These results are discussed in the context of the evolution of alternative mating strategies, and mechanisms of postcopulatory sexual selection.Communicated by N. Wedell     

The origin of the chemical profiles of fungal symbionts and their significance for nestmate recognition in <Emphasis Type="Italic">Acromyrmex</Emphasis> leaf-cutting ants

Cuticular hydrocarbon profiles are essential for nestmate recognition in insect societies, and quantitative variation in these recognition cues is both environmentally and genetically determined. Environmental cues are normally derived from food or nest material, but an exceptional situation may exist in the fungus-growing ants where the symbiotic fungus garden may be an independent source of recognition compounds. To investigate this hypothesis, we quantified the chemical profiles of the fungal symbionts of 18 sympatric colonies of Acromyrmex echinatior and Acromyrmex octospinosus and evaluated the quantitative variation of the 47 compounds in a multivariate analysis. Colony-specific chemical profiles of fungal symbionts were highly distinct and significantly different between the two ant species. We also estimated the relative genetic distances between the fungal symbionts using amplified fragment length polymorphism (AFLP) and correlated these with the overall (Mahalanobis) chemical distances between the colony-specific profiles. Despite the standardized laboratory conditions, the correlations were generally weak, but a statistically significant portion of the total variation in chemical profiles could be explained by genetic differences between the fungal symbionts. However, there was no significant effect of ant species in partial analyses because genetic differences between symbionts tend to coincide with being reared by different ant species. However, compound groups differed significantly with amides, aldehydes, and methyl esters contributing to the correlations, but acetates, alkanes, and formates being unrelated to genetic variation among symbionts. We show experimentally that workers that are previously exposed to and fed with the fungal symbiont of another colony are met with less aggression when they are later introduced into that colony. It appears, therefore, that fungus gardens are an independent and significant source of chemical compounds, potentially contributing a richer and more abundant blend of recognition cues to the colony “gestalt” than the innate chemical profile of the ants alone. Freddie-Jeanne Richard and Michael Poulsen contributed equally to this work.     

The attractiveness fragment—AFLP analysis of local adaptation and sexual selection in a caeliferan grasshopper, <Emphasis Type="Italic">Chorthippus biguttulus</Emphasis>

Genetic variability among males is a necessary precondition for the evolution of female choice based on indirect genetic benefits. In addition to mutations and host–parasite cycles, migration of locally adapted individuals offers an explanation for the maintenance of genetic variability. In a previous study, conducting a reciprocal transplant experiment on a grasshopper, Chorthippus biguttulus, we found that environmental conditions significantly influenced not only body condition but also an important trait of male calling song, the amplitude of song. Although not significant, all other analysed physical and courtship song traits and attractiveness were superior in native than in transferred males. Thus, we concluded that local adaptation has a slight but consistent influence on a range of traits in our study populations, including male acoustic attractiveness. In our present study, we scanned male grasshoppers from the same two populations for amplification fragment length polymorphism (AFLP) loci connected with acoustic attractiveness to conspecific females. We found greater differences in allele frequencies between the two populations, for some loci, than are expected from a balance between drift and gene flow. These loci are potentially connected with locally adapted traits. We examined whether these alleles show the proposed genotype environment interaction by having different associations with attractiveness in the two populations. One locus was significantly related to sexual attractiveness; however, this was independent of the males’ population affiliation. Future research on the evolution of female choice will benefit from knowledge of the underlying genetic architecture of male traits under intraspecific sexual selection, and the ‘population genomics’ approach can be a powerful tool for revealing this structure.     

Investigation of the phylogenetic relationships between <Emphasis Type="Italic">Diuris fragrantissima</Emphasis> and its closest relatives using AFLPs

This study investigated the genetic relationships between the rare terrestrial orchid Diuris fragrantissima and three closely related purple flowered species using Amplified Fragment Length Polymorphisms. The purple-flowered species were found to form individual phenetic clusters confirming recognition of D. fragrantissima at the rank of species. Levels of genetic variation were lower within D. fragrantissima than within its closest relatives but were higher than expected for severely bottlenecked species. Gene flow was detected among all species (Nm = 2.34) and populations (Nm = 0.65) however low among population gene flow indicates that populations may be at risk of divergence. This study has implications for the conservation of D. fragrantissima.     

链格孢菌菌株致病性及其遗传差异性

对分离自紫茎泽兰的19个链格孢菌菌株的致病性进行了比较研究，并对这19个菌株和其它来源的3个菌株的5．8S rDNA及其两侧的ITS1区和ITS2区进行了序列分析．结果显示，链格孢菌菌株的致病性差异明显；21株链格孢菌菌株在5．8SrDNA及其两侧的转录问区（ITS区）的序列无差异，而与百日草链格孢菌（Alternaria zinnine）的序列差异显著，表明21株链格孢菌菌株属种内变异．采用AFLP分子标记技术对21株链格孢菌的DNA扩增片段长度多态性进行了分析，结果表明，链格孢菌自然变异群体内存在很高的遗传多样性．链格孢菌菌株的相似性与菌株的地理来源有一定的相关性，但基于AFLP标记划分的AFLP类群与基于寄主病情指数划分的致病力类群问关系不相一致；起始菌株与变异菌株相似性显著，但二者和致病性的关系不显著．图2表1参18     

Genetic Identification of Spotted Owls, Barred Owls, and Their Hybrids: Legal Implications of Hybrid Identity

Abstract:  Recent population expansion of Barred Owls (  Strix varia ) into western North America has led to concern that they may compete with and further harm the Northern Spotted Owl (  S. occidentalis caurina ), which is already listed as threatened under the U.S. Endangered Species Act (ESA). Because they hybridize, there is a legal need under the ESA for forensic identification of both species and their hybrids. We used mitochondrial control-region DNA and amplified fragment-length polymorphism (AFLP) analyses to assess maternal and biparental gene flow in this hybridization process. Mitochondrial DNA sequences (524 base pairs) indicated large divergence between Barred and Spotted Owls (13.9%). Further, the species formed two distinct clades with no signs of previous introgression. Fourteen diagnostic AFLP bands also indicated extensive divergence between the species, including markers differentiating them. Principal coordinate analyses and assignment tests clearly supported this differentiation. We found that hybrids had unique genetic combinations, including AFLP markers from both parental species, and identified known hybrids as well as potential hybrids with unclear taxonomic status. Our analyses corroborated the findings of extensive field studies that most hybrids genetically sampled resulted from crosses between female Barred Owls and male Spotted Owls. These genetic markers make it possible to clearly identify these species as well as hybrids and can now be used for research, conservation, and law enforcement. Several legal avenues may facilitate future conservation of Spotted Owls and other ESA-listed species that hybridize, including the ESA similarity-of-appearance clause (section 4[e]) and the Migratory Bird Treaty Act. The Migratory Bird Treaty Act appears to be the most useful route at this time.