Molecular Markers For Genetic Analysis Research Articles

Molecular markers for genetic analysis of Indian tropical tasar ecoraces and hybrid populations (Antheraea mylitta.d)

The tropical tasar silkworm Antheraea mylitta D., a wild sericigenous and polyphagous insect is cultivated in the dense, humid, tropical forests of eastern, central and southern India. Its primary food plants are Terminalia arjuna, Terminalia tomentosa and Shorea robusta. The improved varieties of these silkworms can be evolved by hybridization through backcross method, to improve the traits in the upcoming generation by selective parental selection. The present studies of genetic relations between Andhra local and Daba TV ecoraces is identified by DNA markers viz., simple sequence repeats (SSRs), most of which have produced polymorphic bands and based on phylogeny of tasar ecoraces using SSR markers, which further provides molecular evidence of the fact that climatic factors, the changes at DNA level and its wide range of distribution in varied geographic conditions would lead to genetic divergence ultimately leading to the formation of new ecorace.. KEYWORDS :Antheraea mylitta, tasar ecorace, microsatellites, phylogeny, genetic diversity, Breeding, Backcross

Population genetic analysis based on the polymorphisms mediated by transposons in the genomes of pig.

Transposable elements (TEs) mobility is capable of generating a large number of structural variants (SVs), which can have considerable potential as molecular markers for genetic analysis and molecular breeding in livestock. Our results showed that the pig genome contains mainly TE-SVs generated by short interspersed nuclear elements (51,873/76.49%), followed by long interspersed nuclear elements (11,131/16.41%), and more than 84% of the common TE-SVs (Minor allele frequency, MAF > 0.10) were validated to be polymorphic. Subsequently, we utilized the identified TE-SVs to gain insights into the population structure, resulting in clear differentiation among the three pig groups and facilitating the identification of relationships within Chinese local pig breeds. In addition, we investigated the frequencies of TEs in the gene coding regions of different pig groups and annotated the respective TE types, related genes, and functional pathways. Through genome-wide comparisons of Large White pigs and Chinese local pigs utilizing the Beijing Black pigs, we identified TE-mediated SVs associated with quantitative trait loci and observed that they were mainly involved in carcass traits and meat quality traits. Lastly, we present the first documented evidence of TE transduction in the pig genome.

Two new SINE insertion polymorphisms in pig Vertnin (VRTN) gene revealed by comparative genomic alignment

Despite one SINE retrotransposon insertion polymorphism (sRTIP) in the vertebrae development-associated (VRTN) gene was identified in pigs, the structural variations (SVs) in VRTN gene and its proximal flank regions were largely unknown. VRTN genic and flanking sequences from 14 breeds were assembled or downloaded from whole-genome shotgun contings (WGS) database, and aligned to identify the SVs with Clustalx, and retrotransposons in VRTN gene were annotated by RepeatMasker, the splicing patterns of VRTN gene were predicted by Genescan, and large SVs were evaluated by PCR. A total of 12 small SVs and three large SVs in intron of VRTN, derived from SINE insertion polymorphisms, were identified, and two of them (VRTN-sRTIP2 and VRTN-sRTIP3) were not reported before. These VRTN-sRTIPs may affect the splicing patterns of VRTN. They displayed polymorphisms in most detected eight breeds. VRTN-sRTIP2 and VRTN-sRTIP3 showed Hardy-Weinberg equilibrium distributions in most populations except the Chinese local Erhualian pigs, while VRTN-sRTIP1 showed genetic equilibrium in Erhualian pigs. Three VRTN-sRTIPs were identified, and displayed polymorphisms in pigs, and two of them were not reported before. These SVs provide a useful molecular markers for genetic analysis in pigs, and offer new information to facilitate the understanding the SVs of VRTN gene and their putative roles in the variation of vertebral number.

Molecular dating of phylogenetic divergence between Urochloa species based on complete chloroplast genomes

BackgroundForage species of Urochloa are planted in millions of hectares of tropical and subtropical pastures in South America. Most of the planted area is covered with four species (U. ruziziensis, U. brizantha, U. decumbens and U. humidicola). Breeding programs rely on interspecific hybridizations to increase genetic diversity and introgress traits of agronomic importance. Knowledge of phylogenetic relationships is important to optimize compatible hybridizations in Urochloa, where phylogeny has been subject of some controversy. We used next-generation sequencing to assemble the chloroplast genomes of four Urochloa species to investigate their phylogenetic relationships, compute their times of divergence and identify chloroplast DNA markers (microsatellites, SNPs and InDels).ResultsWhole plastid genome sizes were 138,765 bp in U. ruziziensis, 138,945 bp in U. decumbens, 138,946 bp in U. brizantha and 138,976 bp in U. humidicola. Each Urochloa chloroplast genome contained 130 predicted coding regions and structural features that are typical of Panicoid grasses. U. brizantha and U. decumbens chloroplast sequences are highly similar and show reduced SNP, InDel and SSR polymorphism as compared to U. ruziziensis and U. humidicola. Most of the structural and sequence polymorphisms were located in intergenic regions, and reflected phylogenetic distances between species. Divergence of U. humidicola from a common ancestor with the three other Urochloa species was estimated at 9.46 mya. U. ruziziensis, U. decumbens, and U. brizantha formed a clade where the U. ruziziensis lineage would have diverged by 5.67 mya, followed by a recent divergence event between U. decumbens and U. brizantha around 1.6 mya.ConclusionLow-coverage Illumina sequencing allowed the successful sequence analysis of plastid genomes in four species of Urochloa used as forages in the tropics. Pairwise sequence comparisons detected multiple microsatellite, SNP and InDel sites prone to be used as molecular markers in genetic analysis of Urochloa. Our results placed the origin of U. humidicola and U. ruziziensis divergence in the Miocene-Pliocene boundary, and the split between U. brizantha and U. decumbens in the Pleistocene.

SNP discovery from next-generation transcriptome sequencing data and their validation using KASP assay in wheat (Triticum aestivum L.)

Single nucleotide polymorphisms (SNPs) are becoming the most amenable form of DNA-based molecular markers for genetic analysis. In hexaploid bread wheat (Triticum aestivum L.), it is difficult to discern true polymorphic SNPs due to homoeologous and paralogous genes. Two serial analysis of gene expression (SAGE) libraries were developed utilizing leaves from resistant plants carrying leaf rust resistance gene Lr28; one library was derived from leaves that were mock inoculated and the other was derived from leaves inoculated with the urediniospores of the leaf rust pathogen Puccinia triticina. Next-generation sequencing reads, after quality trimming and removal of fungal sequences, were mapped to wheat reference sequences at Ensembl Plants. CLC Genomics Workbench and Freebayes softwares were employed for SNP calling. A total of 611 SNPs were predicted to be common by both softwares, of which 207 varietal SNPs were identified by ConservedPrimer software. A subset of 100 SNPs was used for validation across 47 wheat genotypes using Kompetitive Allele Specific PCR (KASP) assay; 83 SNPs could be successfully validated. These SNPs were positioned on wheat subgenomes and chromosome arms. When functionally annotated, many sequences harboring SNPs showed homology to resistance and resistance-like genes listed in Plant Resistance Gene database (PRGdb) as well as pathogenesis-related (PR) and stress-responsive genes. The results of the present study involving discovery of SNPs associated with resistance to leaf rust, a major threat to wheat production worldwide, will be valuable for molecular breeding for rust resistance.

A REVIEW OF THE INVASIVE EURASIAN COLLARED DOVE AND POSSIBLE RESEARCH METHODS IN THE FUTURE

The Eurasian Collared Dove Streptopelia decaocto is one of the most successfuly expanding vertebrate species in the world; therefore, it has attracted special scientific focus. This review summarises the species’ achievements related to its rapid expansion and the ecological consequences with regard to the population genetic assessments. It also highlights the expansion dynamics of the species and the ecosystems to which it is most adapted. Finally, the effects of this species on the newly colonised ecosystems are illustrated. The study of the genetic structure between populations to make inferences about expansion dynamics has been neglected so far. Thus, we survey the opportunities of using molecular markers for genetic analysis in populations of Eurasian Collared Dove as a possible means for future studies. This work is important given the lack of information on the mentioned fields, and it calls attention to the importance of further interdisciplinary studies from different scientific fields using this species. .

Establishment and Optimization of Genomic Selection to Accelerate the Domestication and Improvement of Intermediate Wheatgrass.

Intermediate wheatgrass (IWG) is a perennial species and has edible and nutritious grain and desirable agronomic traits, including large seed size, high grain yield, and biomass. It also has the potential to provide ecosystem services and an economic return to farmers. However, because of its allohexaploidy and self-incompatibility, developing molecular markers for genetic analysis and molecular breeding has been challenging. In the present study, using genotyping-by-sequencing (GBS) technology, 3436 genome-wide markers discovered in a biparental population with 178 genets, were mapped to 21 linkage groups (LG) corresponding to 21 chromosomes of IWG. Genomic prediction models were developed using 3883 markers discovered in a breeding population containing 1126 representative genets from 58 half-sib families. High predictive ability was observed for seven agronomic traits using cross-validation, ranging from 0.46 for biomass to 0.67 for seed weight. Optimization results indicated that 8 to 10 genets from each half-sib family can form a good training population to predict the breeding value of their siblings, and 1600 genome-wide markers are adequate to capture the genetic variation in the current breeding population for genomic selection. Thus, with the advances in sequencing-based marker technologies, it was practical to perform molecular genetic analysis and molecular breeding on a new and challenging species like IWG, and genomic selection could increase the efficiency of recurrent selection and accelerate the domestication and improvement of IWG.

CandiSSR: An Efficient Pipeline used for Identifying Candidate Polymorphic SSRs Based on Multiple Assembled Sequences.

Simple sequence repeats (SSRs), also known as microsatellites, are ubiquitous short tandem duplications commonly found in genomes and/or transcriptomes of diverse organisms. They represent one of the most powerful molecular markers for genetic analysis and breeding programs because of their high mutation rate and neutral evolution. However, traditionally experimental screening of the SSR polymorphic status and their subsequent applicability to genetic studies are extremely labor-intensive and time-consuming. Thankfully, the recently decreased costs of next generation sequencing and increasing availability of large genome and/or transcriptome sequences have provided an excellent opportunity and sources for large-scale mining this type of molecular markers. However, current tools are limited. Thus we here developed a new pipeline, CandiSSR, to identify candidate polymorphic SSRs (PolySSRs) based on the multiple assembled sequences. The pipeline allows users to identify putative PolySSRs not only from the transcriptome datasets but also from multiple assembled genome sequences. In addition, two confidence metrics including standard deviation and missing rate of the SSR repetitions are provided to systematically assess the feasibility of the detected PolySSRs for subsequent application to genetic characterization. Meanwhile, primer pairs for each identified PolySSR are also automatically designed and further evaluated by the global sequence similarities of the primer-binding region, ensuring the successful rate of the marker development. Screening rice genomes with CandiSSR and subsequent experimental validation showed an accuracy rate of over 90%. Besides, the application of CandiSSR has successfully identified a large number of PolySSRs in the Arabidopsis genomes and Camellia transcriptomes. CandiSSR and the PolySSR marker sources are publicly available at: http://www.plantkingdomgdb.com/CandiSSR/index.html.

DKRE1—The first full-length Ty1-copia-like retrotransposon in persimmon: Isolation, characteristic and potential involvement in occurrence of bud mutations

The focus of this study revolved around persimmon (Diospyros kaki Thunb.) retrotransposable element 1 (DKRE1), a complete long terminal repeat (LTR) retrotransposon (RTN) that was isolated from persimmons. Its structure, phylogenetic classification, copy number, active characteristics and potential roles involved in the occurrence of bud mutations in persimmons were investigated. Having a length of 6, 235bp, DKRE1 possessed all of the sequence features associated with Ty1-copia-like retrotransposons. Phylogenetic analysis showed significant, but not strong, identity in amino acid sequences with BARE1 (barley retrotransposable element-1), RIRE1 (rice retrotransposable element-1) and Ppcrt (Pyrus pyrifolia copia retrotransposon) groups. The estimated copy number of 45 copies per haploid genome suggested that DKRE1 possesses a low constitution in the persimmon genome. Furthermore, the transcription of DKRE1 could be detected in leaf tissue treated with salicylic acid (SA), methyl jasmonate (MJA), and abscisic acid (ABA), indicating its active characteristics when subjected to exogenous phytohormones. Polymorphisms were revealed between several pairs of bud mutations by way of either inter-retrotransposon amplified polymorphism (IRAP) or sequence-specific amplified polymorphism (SSAP) retrotransposon-based molecular markers, showing a potential involvement in their occurrence. To our knowledge, this is the first report of the isolation of a complete LTR retrotransposon with transcriptional activity in persimmons. This study is valuable for future research on the mechanisms of bud mutation and development of retrotransposon-based molecular markers for genetic analysis.

Use IRAP and IPBS molecular markers for genetic analysis of wheat varieties

Use IRAP and IPBS molecular markers for genetic analysis of wheat varieties

Brassica A genome의 최근 연구 동향

작물의 구조와 기능을 이해하려는 과학적 탐구심과 이를 작물 육종에 적용하려는 실험적 노력의 일환으로 다양한 작물에서 유전자 지도가 개발되었다. 특히, 배추과 작물의 경우 모델식물인 애기장대의 유전체 정보가 공개된 이후 다양한 정보 (염기서열 정보, 유전자 구조 및 기능정보 등)의 이용이 가능해져 유전자 지도 작성이 가속화 되었으며 이는 최근 <TEX>$B.$</TEX> <TEX>$rapa$</TEX> A genome (배추)유전체 해독이라는 결과를 가져왔다. 배추과 작물의 유전자 지도 작성에 있어서 초기에는 RFLP 마커들이 사용되었으나 이후 분자마커, 즉, RAPD, AFLP, SSR 등과 같이 비교적 사용이 간단하고 시간적 제약이 없는 PCR 마커의 형태로 점차 바뀌었다. 배추과 작물의 경제적, 학문적 가치가 고려되어 <TEX>$B.$</TEX> <TEX>$rapa$</TEX> (배추)를 표준재료로 A genome 유전체 염기서열 해독이라는 목표로 다국적 유전체 프로젝트가 결성되었고 2011년 국내연구진이 주도적으로 참여한 국제 컨소시엄 (BrGSPC, <TEX>$B.$</TEX> <TEX>$rap</TEX><TEX>a$</TEX> Genome Sequencing Project Consortium)에 의해 배추 (10개 염색체)의 유전자 영역(gene space), 약 98% (83.8 Mb)의 염기서열이 해독되어 발표되었다. 유전체 해독 과정에서 축적된 염기서열 정보는 대량의 SSR, SNP, IBP 마커의 개발을 가능하게 하였고 이들 마커는 <TEX>$B.$</TEX> <TEX>$rapa$</TEX> A genome 유전자 지도와 물리 지도 작성에 이용되어 이후 배추과 작물연구 전반에 널리 적용되고 있다. 대량의 분자마커 개발은 유전자 지도 작성을 가속화하여 더욱 정밀한 유전자 지도를 가능하게 하였고 공통의 분자마커 정보는 애기장대와 배추과 작물 간 비교유전체 연구를 통해 농업적 우수 형질의 클로닝, 마커도움선발 (MAS)등의 방법으로 분자육종의 기반을 제공하고 있다. 뿐만 아니라. 최근 등장한 NGS 유전체 해독 기술로 생산된 대량의 정보는 분자육종 실현 가능성을 높여 분자육종 실용화에 박차를 가하는 계기가 되고 있다. 본 논문에서는 <TEX>$B.$</TEX> <TEX>$rapa$</TEX>에서 분자마커를 이용한 유전자 지도 개발의 과정과 농업적 유용형질 탐색을 위한 양적 형질 유전자좌 (QTLs)의 연구 현황에 대하여 알아보고 유전체연구에서 유전자 지도의 중요성과 육종에의 응용에 대하여 서술하였다. 또한 다양한 유전체 정보와 오믹스 정보를 국내 배추과 분자육종에 효율적으로 활용하여 분자육종 실용화를 가능하게 하기 위해 사용자가 쉽게 사용할 수 있는 데이터베이스를 구축함으로서 연구자와 육종가 간의 간격을 좁히고 원활한 정보교환의 필요성을 제기하였다. As a scientific curiosity to understand the structure and the function of crops and experimental efforts to apply it to plant breeding, genetic maps have been constructed in various crops. Especially, in the case of Brassica crop, genetic mapping has been accelerated since genetic information of model plant <TEX>$Arabidopsis$</TEX> was available. As a result, the whole <TEX>$B.$</TEX> <TEX>$rapa$</TEX> genome (A genome) sequencing has recently been done. The genome sequences offer opportunities to develop molecular markers for genetic analysis in <TEX>$Brassica$</TEX> crops. RFLP markers are widely used as the basis for genetic map construction, but detection system is inefficiency. The technical efficiency and analysis speed of the PCR-based markers become more preferable for many form of <TEX>$Brassica$</TEX> genome study. The massive sequence informative markers such as SSR, SNP and InDels are also available to increase the density of markers for high-resolution genetic analysis. The high density maps are invaluable resources for QTLs analysis, marker assisted selection (MAS), map-based cloning and comparative analysis within <TEX>$Brassica$</TEX> as well as related crop species. Additionally, the advents of new technology, next-generation technique, have served as a momentum for molecular breeding. Here we summarize genetic and genomic resources and suggest their applications for the molecular breeding in <TEX>$Brassica$</TEX> crop.

Linkage disequilibrium, genetic association mapping and gene localization in crop plants

DNA-based molecular markers have been extensively utilized for a variety of studies in both plant and animal systems. One of the major uses of these markers is the construction of genome-wide molecular maps and the genetic analysis of simple and complex traits. However, these studies are generally based on linkage analysis in mapping populations, thus placing serious limitations in using molecular markers for genetic analysis in a variety of plant populations. Therefore, alternative approach has been suggested, linkage disequilibrium-based association analysis which detects and locates quantitative trait loci (QTL) by the strength of the correlation between a trait and a marker. Although association analysis has already been used for studies on genetics of complex traits in humans, its use in plants has newly started. In the present review, we describe what is known about variation in linkage disequilibrium (LD) and summarize published results on association studies in crop plant species. We give a list of different factors affecting LD, and discuss the current issues of LD research in plants. Later, we also describe the various uses of LD in crop plants research and summarize the present status of LD researches in different plant genomes. Finally, future key issues about the application of these studies on the localization of genes in these crop plants have been also discussed.

Linkage disequilibrium and association studies in higher plants: Present status and future prospects

During the last two decades, DNA-based molecular markers have been extensively utilized for a variety of studies in both plant and animal systems. One of the major uses of these markers is the construction of genome-wide molecular maps and the genetic analysis of simple and complex traits. However, these studies are generally based on linkage analysis in mapping populations, thus placing serious limitations in using molecular markers for genetic analysis in a variety of plant systems. Therefore, alternative approaches have been suggested, and one of these approaches makes use of linkage disequilibrium (LD)-based association analysis. Although this approach of association analysis has already been used for studies on genetics of complex traits (including different diseases) in humans, its use in plants has just started. In the present review, we first define and distinguish between LD and association mapping, and then briefly describe various measures of LD and the two methods of its depiction. We then give a list of different factors that affect LD without discussing them, and also discuss the current issues of LD research in plants. Later, we also describe the various uses of LD in plant genomics research and summarize the present status of LD research in different plant genomes. In the end, we discuss briefly the future prospects of LD research in plants, and give a list of softwares that are useful in LD research, which is available as electronic supplementary material (ESM).

Global genetic analysis.

The introduction of molecular markers in genetic analysis has revolutionized medicine. These molecular markers are genetic variations associated with a predisposition to common diseases and individual variations in drug responses. Identification and genotyping a vast number of genetic polymorphisms in large populations are increasingly important for disease gene identification, pharmacogenetics and population-based studies. Among variations being analyzed, single nucleotide polymorphisms seem to be most useful in large-scale genetic analysis. This review discusses approaches for genetic analysis, use of different markers, and emerging technologies for large-scale genetic analysis where millions of genotyping need to be performed.

A comparison of molecular markers for genetic analysis of macadamia

SummaryVarious marker systems exist for genetic analysis of horticultural species. Isozymes were first applied to the woody perennial nut crop, macadamia, in the early 1990s. The advent of DNA markers saw the development, for macadamia, of STMS (sequence-tagged microsatellite site), RAPD (randomly amplified polymorphic DNA), and RAF (randomly amplified DNA fingerprinting). The RAF technique typically generates dominant markers, but within the dominant marker profiles, certain primers also amplify multi-allelic co-dominant markers that are suspected to be microsatellites. In this paper, we confirm this for one such marker, and describe how RAF primers can be chosen that amplify one or more putative microsatellites. This approach of genotyping anonymous microsatellite markers via RAF is designated RAMiFi (randomly amplified microsatellite fingerprinting). Several marker systems were compared for the type, amount, and cost-efficiency of the information generated, using data from published studies on macadamia. The markers were also compared for the way they clustered a common set of accessions. The RAMiFi approach was identified as the most efficient and economical. The availability of such a versatile tool offers many advantages for the genetic characterisation of horticultural species.

Identification of SNPs in human gamma aminobutyric acid A receptor γ2 gene

Gamma aminobutyric acid (GABA) is a major inhibitory neurotransmitter. Because of its importance and diverse functions, studies of single nucleotide polymorphisms in the GABA receptor genes are of great significance, which may produce useful molecular markers for genetic analysis of neurological disease, and possibly lead to the discovery of pathogenic mutations. We identified eleven SNPs in the entire exon regions, immediate intron regions and promoter region of human GABA receptor gamma2 gene by direct sequencing. In this discovery, 2 non-synonymous cSNPs and 1 splicing variant are found.

Mapping of the OB receptor to 1p in a region of nonconserved gene order from mouse and rat to human.

As part of an effort to identify informative molecular markers for genetic analysis of human pedigrees segregating for obesity, we have developed a genetic map of human 1p in the region of the OB receptor (OBR), the gene that is defective in murine diabetes (Obrdb) and rat Zucker fatty (Obrfa) mutations located on mid-chromosome 4 and chromosome 5, respectively. OBR was mapped 0.9 cR centromeric to WI-9515 and 2.2 cR telomeric of WI-7249 by radiation hybrid (RH) mapping. Ten yeast artificial chromosomes (YACs) containing OBR were identified, confirming the location of OBR centromeric to WI-9515 and telomeric to WI-7249. Additionally, five P1 artificial chromosomes (PACs) were identified that comprised a contiguous series of overlapping clones spanning the length of OBR. WI-5182 was contained within the two PACs that are 3' of OBR. Using a panel of 68 individuals from a single three-generation family and an additional nuclear family, we have mapped 18 polymorphic markers including phosphoglucomutase 1 (PGM1), which is centromeric to Obrdb / Obrfa, and D1S85, which is telomeric to Obrdb / Obrfa in the mouse and rat. The following composite map integrates these radiation hybrid, genetic, and physical maps: Centromere-@WI-7249-[OBR; WI-5182]-D1S198-[WI-9515; WI-6550; D1S2866]-D1S2825-[WI-3077; D1S2886]-[D1S515; DS1613; PGM1]-[D1S312; D1S473; D1S230; D1S246; D1S203]-D2S1643-[D1S1669; D1S1596;]UNCJ-D1S476- D1S85-D1S220-C8B-GTAT1A7. Unresolvable markers are within brackets. A comparison of gene order on mouse chromosome 4, rat chromosome 5, and human 1p indicates that between rodents and humans, there has been a rearrangement of the gene order in the region surrounding OBR.

Molecular Markers in Genetic Analysis of Tomato

ABSTRACTTomato is among the first species to which molecular markers are applied extensively for genomic studies, tagging and map-based cloning of genes, marker-assisted selection and phylogenetic studies. More than 1000 RELP markers build a high density molecular linkage map that provides molecular access to any chromosomal region of interest. PCR techniques supply breeding programmes with additional markers (RAPDs and SCARs) which are suitable for diagnostic purposes by their virtue of being fast, safe and easy to automate By combining the RAPD assay with proper screening systems involving NILs or pools of genotypes, markers tigthly linked to a gene of interest can be identified which fapihtaie chromosome walking. Thus, molecular marker technology is bringing about a revolution in the practice of molecular genetics and plant breeding.

Potential applications of molecular markers for genetic analysis of host–pathogen systems in forest trees

Applications of molecular markers for genetic analysis of host–pathogen systems are presented within the framework of the gene-for-gene model. The literature on complementary genetic interactions in host–pathogen systems is briefly reviewed. Flor's gene-for-gene hypothesis is summarized, and the design, execution, and genetic interpretation of experiments to test this hypothesis are described in detail. Various molecular techniques that have been used to complement the traditional gene-for-gene approach are also briefly reviewed. Three alternative approaches to developing molecular markers using the gene-for-gene framework are presented to illustrate the potential for applying molecular markers in forest tree–pathogen systems. Two of these alternative approaches allow partial confirmation of a hypothesized gene-for-gene genetic model without the usual requirement for advanced pedigrees. Such alternative approaches could greatly accelerate the development of marker-aided selection for disease resistance in forest tree–pathogen systems.

Molecular Markers for Genetic Analysis of Phytopathogenic Fungi

Genetic studies of any organism require precise, easily scored characters or markers. In the past, naturally occurring markers were often insufficient and mutagenesis was used to generate auxotrophic, morphological, or drug­ resistant mutants. These techniques have been extensively exploited to devel­ op several Ascomycete fungi (Saccharomyces cerevisiae, Aspergillus nidu­ lans, and Neurospora crassa) as model systems for rigorous genetic analysis (42). Despite the early demonstration of the Mendelian inheritance of genes determining virulence, progress in understanding the genetics of phyto­ pathogenic fungi has lagged behind other eukaryotes due to the difficulties of manipulation and the lack of suitable markers. Recently molecular techniques have provided two classes of markers that will allow extensive analysis of phytopathogenic fungi. Isozymes have been extensively used in several plant and animal species and are beginning to be employed with phytopathogenic fungi (13). DNA restriction fragment length polymorphism (RFLP) analyses are extensively used in human genetics, and are becoming routine with some plants (5, 66, 98, 103), but are in their infancy for phytopathogenic fungi. Molecular markers have several advan­ tages over other types of markers: (a) numerous molecular markers can be identified; (b) most are codominant (i.e. heterozygotes can be distinguished from homozygotes), allowing the identification of all possible genotypes; (c) they do not usually have deleterious or strong epistatic effects, allowing many markers to be considered simultaneously; (d) genotypes of molecular loci can often be determined from any tissue; and (e) numerous alleles may exist for some markers. In addition, RFLP markers may act as starting points for