INTROGRESSING PHEROMONE QTL BETWEEN SPECIES: TOWARDS AN EVOLUTIONARY UNDERSTANDING OF DIFFERENTIATION IN SEXUAL COMMUNICATION

As a first step toward understanding how noctuid moths evolve species-specific pheromone communication systems, we hybridized and backcrossed two closely related moth species, Heliothis virescens (Hv) and H. subflexa (Hs), which differ qualitatively and quantitatively in their multi-component sex pheromone blends. We used amplified fragment length polymorphism (AFLP) marker-based mapping of backcross families to determine which of the 30 autosomes in these moths contained quantitative trait loci (QTL) controlling the percentages of specific chemical components in the pheromone blends. In two previous backcrosses to Hs, we found a strong depressive effect of Hv-chromosome 22 on the percentage of three acetate components in the pheromone gland. These acetates are present in Hs and absent in Hv. Here, we describe how we introgressed Hv-chromosome 22 into the genomic background of Hs. Selection for Hv-chromosome 22 started from backcross 3 (BC₃) females. All females that had Hv-chromosome 22 and a low percentage of acetates (< 3% of the total amount of pheromone components present) were backcrossed to Hs males. In BC₅ to BC₈, we determined whether Hv-chromosome 22 was present by a) running only the primer pairs that would yield the markers for that chromosome, and/or b) determining the relative percentages of acetates in the pheromone glands. Either or both genotype and phenotype were used as a criterion to continue to backcross these females to Hs males. In BC₉, we confirmed the isolation of Hv-chromosome 22 in the Hs genomic background, and backcrossed the males to Hs females to eliminate the Hv-sex chromosome as well as mitochondrial DNA. The pheromone composition was determined in BC₃, BC₅, and BC₁₁ females with and without Hv-chromosome 22. All backcross females with Hv-chromosome 22 contained significantly less acetates than females without this chromosome. In addition, BC₃ females with Hv-chromosome 22 contained significantly more Z11-16:OH than BC₃ females without Hv-chromosome 22. However, in BC₅ and BC₁₁ females, the correlation between Z11-16:OH and Hv-chromosome 22 was lost, suggesting that there are separate QTL for the acetates and for Z11-16:OH, and that the relative amount of the alcohol component is only affected in epistasis with other (minor) QTL. Now that we have succeeded in isolating the chromosome that has a major effect on acetate production, we can test in behavioral experiments whether the presence of acetates may have been a driving force for a shift in pheromone composition. Such tests are necessary to move towards an evolutionary understanding of the differentiation in sexual communication in Heliothis spp. moths.     

Application of molecular identification tools for Lactobacillus, with a focus on discrimination between closely related species: A review

Lactobacillus is among the most important GRAS food lactic acid bacteria, with nearly 140 species at present, mostly of industrial importance. Being part of the natural flora of a range of food products like raw milk, fermented dairy products, fruits, vegetables, meat products they also serve as starters for a number of fermented food products either to enhance the quality or to add health benefits. These groups of economically important species are often alike in phenotypic and physiological characteristics, probably due to their co-evolution in the same ecological niches; hence they are difficult to be differentiated. This demands advanced methods for their proper identification and characterization. With the advancement of molecular biology, a range of DNA-based molecular techniques has replaced the largely cumbersome phenotypic methods. This review summarizes the various molecular techniques available for detection and identification within the genus Lactobacillus, with special emphasis on the four groups of closely resembling species: L. casei group, L. acidophilus group, L. delbrueckii subspecies, and L. plantarum group. This review also provides insights into current trends for alternative molecular markers other than 16S rRNA to resolve the ambiguity within phylogenetically close species in the genus Lactobacillus.     

Genetic Differentiation between Natural and Hatchery Stocks of Japanese Scallop (Mizuhopecten yessoensis) as Revealed by AFLP Analysis

Japanese scallop (Mizuhopecten yessoensis) is a cold-tolerant bivalve that was introduced to China for aquaculture in 1982. In this study, amplified fragment length polymorphism (AFLP) markers were used to investigate levels of genetic diversity within M. yessoensis cultured stocks and compare them with wild populations. Six pairs of primer combinations generated 368 loci among 332 individuals, in four cultured and three wild populations. High polymorphism at AFLP markers was found within both cultured and wild M. yessoensis populations. The percentage of polymorphic loci ranged from 61.04% to 72.08%, while the mean heterozygosity ranged from 0.2116 to 0.2596. Compared with wild populations, the four hatchery populations showed significant genetic changes, such as lower expected heterozygosity and percentage of polymorphic loci, and smaller frequency of private alleles, all indicative of a reduction in genetic diversity. Some genetic structures were associated with the geographical distribution of samples; with all samples from Dalian and Japan being closely related, while the population from Russia fell into a distinct clade in the phylogenetic analysis. The genetic information derived from this study indicated that intentional or accidental release of selected Japanese scallops into natural sea areas might result in disturbance of local gene pools and loss of genetic variability. We recommend monitoring the genetic variability of selected hatchery populations to enhance conservation of natural Japanese scallop resources.     

Genetic Transformation of Watermelon with Pumpkin DNA by Low Energy Ion Beam-mediated Introduction

The No.601 watermelon (citrullus lanatus) seeds were treated with 25 keV N+ implantation at the dosage of 7.8*10 ions/cm2 . After treatment, watermelon seeds were incubated with 380 *g/*l pumpkin (Cucubita, maxima Duch) DNA solution at 35* for 5 hours. By two-generations of selection and resistance screening at seedling stage, one transformed material was selected out, whose rind color is similar to that of the donor pumpkin and whose size of seeds is between that of the donor and the receptor. Using AFLP (amplified fragment length polymorphism) technique, two polymorphic DNA fragments were amplified. This primarily testified that the donor DNA fragments/gene were introduced into the receptor cell and integrated into the genomic DNA of the receptor.     

应用AFLP建立玉米基因组DNA指纹图谱的研究

分别采用PstI单酶切AFLP和EcoRI/MseI双酶切AFLP建立6个玉米品系的AFLPDNA指纹图谱,并进行亲缘关系鉴定。相比之下,PstI单酶切AFLP法较简单、经济,而EcoRI/MseI双酶切AFLP法扩增出的条带数量多、背景低;二者均适用于亲缘关系鉴定。     

黑龙江省大豆灰斑病菌遗传多样性

从100对AFLP引物组合中筛选出多态性高且稳定性好的10对引物组合,对2006-2009年采自黑龙江省各大豆主要产区的121个大豆灰斑病菌菌株进行AFLP分析,得到148个多态性条带。用UPGMA聚类分析,121个菌株被归类于111个不同单元型（带型）,以彼此间的遗传距离小于0.81为界,将111个单元型划分为6个遗传系谱。其中L03、L05和L06为黑龙江大豆灰斑病的优势系谱。L02分布地域比较狭窄,为黑龙江大豆灰斑病的稀有遗传系谱。     

长江中下游地区6个克氏原螯虾群体遗传多样性分析

为探讨长江中下游地区克氏原螯虾群体遗传多样性,使用AFLP标记,用5个EcoRI—MseI引物从6个群体139个个体中共扩增出129个位点,其中多态性位点为89,占68．99％,通过NTSYSpc软件对统计的I／O矩阵进行分析。结果表明：在群体水平上,洪泽湖显示了最大的遗传多样性,多态位点比例为75．74％,有效等位基因数为1．4181,Nei’S遗传多样性指数为0．5829,Shannon多样性信息指数为0．441;群体问发生了较高程度的遗传分化,6个群体问的遗传分化系数为0．4186,群体的遗传变异91．20％来源于群体内,8．80％来源于群体间,遗传变异主要存在于群体内,群体内的遗传分化大于群体间的分化;6个克氏原螯虾群体可以分为3组,而鄱阳湖群体在这3组中均有分布。     

Metabolic footprinting as a tool for discriminating between brewing yeasts

The characterization of industrial yeast strains by examining their metabolic footprints (exometabolomes) was investigated and compared to genome-based discriminatory methods. A group of nine industrial brewing yeasts was studied by comparing their metabolic footprints, genetic fingerprints and comparative genomic hybridization profiles. Metabolic footprinting was carried out by both direct injection mass spectrometry (DIMS) and gas chromatography time-of-flight mass spectrometry (GC-TOF-MS), with data analysed by principal components analysis (PCA) and canonical variates analysis (CVA). The genomic profiles of the nine yeasts were compared by PCR-restriction fragment length polymorphism (PCR-RFLP) analysis, genetic fingerprinting using amplified fragment length polymorphism (AFLP) analysis and microarray comparative genome hybridizations (CGH). Metabolomic and genomic analysis comparison of the nine brewing yeasts identified metabolomics as a powerful tool in separating genotypically and phenotypically similar strains. For some strains discrimination not achieved genomically was observed metabolomically.     

利用AFLP技术研究海湾扇贝不同养殖群体的遗传结构及其分化

采用AFIP分子标记技术对我国海湾扇贝的4个不同地理群体共80个个体利用7对引物组合进行了遗传多样性分析。4群体的多态位点比例分别为：加拿大养殖群体48．8235％,墨西哥湾养殖群体49．2914％,秦皇岛养殖群体38．2353％,浙江养殖群体41．1765％。平均杂和度分别为0．1878,0．1886,0．1265和0．1618。遗传距离介于0．1188～0．0941之间。4个群体的Fst为0．1883。根据遗传距离绘制了UPGMA聚类图。4个群体间的遗传关系如下：加拿大群体和浙江群体聚为一支,墨西哥湾群体和秦皇岛群体聚成一支,最后这两支聚在一起。试验结果表明,经过长时间累代养殖的浙江、加拿大、墨西哥湾群体仍保持较高的杂合度,与引种时间最短的(2年)、最为接近美国自然群体的秦皇岛群体比较,以上3个群体没有出现多态位点比例和杂合度显著下降的现象。