Vol.7 No.2B June 2002
APPLICATION OF MOLECULAR MARKERS IN STUDIES ON PLANTS
SEPTEMBER 25-29, 2002, WARSAW, POLAND
PART II
| Volume 7 (2002) pp 625-634 | |
| Title | THE USE OF CYTOPLASMIC MARKERS IN ONION HYBRID BREEDING |
| Authors | Marek Szklarczyk*, Magdalena Simlat, Barbara Jagosz and Grażyna Ba |
| Abstract | We applied the RFLP approach to identify the cytoplasmic genotypes of selected onion breeding materials from Poland. For this purpose, mitochondrial DNA from cytoplasmic male-sterile (CMS) and male-fertile onions were hybridized with the probes for the following mitochondrial genes: atpA, atp6, atp9, cob, cox1, nad3, nad4 and nad6. S-, T- or C-cytoplasm was represented in each analyzed sterile accession. Some new polymorphisms shared by S- and C-cytoplasmic onions were identified. We also used currently available PCR markers to test if cytoplasmic heterogenity occurs within onion inbreds. A fraction of the plants bearing S-cytoplasm were found within two male-fertile lines, but such plants were not detected in the open-pollinated cultivars Sochaczewska, Wolska and Żytawska. Both the RFLP and PCR approaches gave some proof of existing mitochondrial heteroplasmy in onions. |
| Address and Contact Information | Department of Genetics, Plant Breeding and Seed Science,
Agricultural University of Kraków, 29 Listopada 54, 31-425 Kraków, Poland * Corresponding author, E-mail: marekszk@hotmail.com |
 ![[Rozmiar: 1332 bajtĂłw]](pic/abstract.gif){kind=small} |
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| Volume 7 (2002) pp 635-648 | |
| Title | MORPHOLOGICAL, CYTOLOGICAL AND BSA-BASED TESTING ON LIMITED SEGREGATION POPULATION AFLPS |
| Authors | Piotr Tomasz Bednarek*, Helena Kubicka and Małgorzata Zawada |
| Abstract | Cytoplasmic male sterility (cms) in rye (Secale cereale L.), especially cytoplasma PAMPA, is used commercially in hybrid breeding programmes. The development of molecular markers that are tightly linked to the numerous genes coding for pollen fertility is expected to have great impact in the field. Morphological and cytological analyses of plants from a three-way cross C394: [(S67P/94 x S38/94) x CHD296] indicated the presence of at least several genes acting at different stages of pollen grain development, and proved the concurrence of both approaches in plant classification. The AFLP technique combined with the Bulk Segregant Analysis (BSA) were applied to identify DNA fragments linked to the genes of interest. All the 256 possible primer pair combinations based on the MseI and EcoRI restriction sites generated distinct band patterns allowing the identification of 31143 DNA fragments, visualised using the isotopic method. On average, any given primer combination generated 122 fragments. Among 1111 and 431 potential genetic markers respectively identified in the restorer form and the maternal lines, 775 and 295 were present in the F2 population. These numbers were then reduced to 109 and 51. The identified DNA fragments were tested on a limited segregating population, C394-F2, in order to eliminate false signals and to select markers for a future marker-assisted selection programme. Twenty-five markers were selected. Four of these markers were not identified via the BSA approach, indicating that if a highly polymorphic component is used for a cross, or a polygenic trait is studied, then the use of a limited population may be required. |
| Address and Contact Information | The Botanical Garden-the Centre for the Conservation of Biological Diversity
of the Polish Academy of Sciences, 02-973 Warsaw, ul. Prawdziwka 2, Poland * Corresponding author : E-mail: tmol.ob@ihar.edu.pl, Fax:+48 22 757 66 45 |
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| Volume 7 (2002) pp 649-655 | |
| Title | IDENTIFICATION OF RAPD MARKERS AND THEIR USE FOR MOLECULAR MAPPING IN PEA (PISUM SATIVUM L.) |
| Authors | Kianoosh Cheghamirza*, Oksana Koveza, Fedor Konovalov and Sergei Gostimsky |
| Abstract | The RAPD method (Random Amplified Polymorphic DNA) was used for identifying and mapping new molecular markers in pea. RAPD analysis of various cultivars and lines of pea was carried out using 10-mer random primers. The presence of multiple polymorphism between cultivars and lines was revealed; at least one fragment for any given primer was present in the DNA of one form of pea and absent in the DNA of another line or cultivar. To detect molecular markers linked to the genes of chi-15, xa-18 and also to the 12 morphological markers of the L-1238 line, the F2 populations (Chi-15 x L-1238), (Vio x L-1238), (Xa-18 x L-1238), (L-111 x Chi-15) and (L-84 x Xa-18) were studied via bulked segregant analysis. DNA molecular analysis of F1 hybrids revealed the presence of parental polymorphic fragments in all of the populations. The study of the F2 plants showed that the obtained fragments are inherited as Mendelian factors. 13 RAPD-markers linked to genes of A/a (flower color), I/i (seed color), Gp/gp (pod color), R/r (seed form), S/s (seeds linkage), and also to genes of Chi-15/chi-15 (leaf color) and Xa-18/xa-18 (leaf color) were discovered. The study of individual plant DNA from the F2 populations allowed us to determine the genetic distances between genes and the RAPD markers linked to them. |
| Address and Contact Information | Genetics and Breeding Department, Moscow State University, Vorobjevy Gory,
Moscow, 119899, Russia *Corresponding author, E-mail: Cheghamirza@yandex.ru |
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| Volume 7 (2002) pp 657-663 | |
| Title | GENETIC ANALYSIS OF POD DEHISCENCE IN PEA (PISUM SATIVUM L.) |
| Authors | Norman F. Weeden1, Soren Brauner2 and Jerzy A. Przyborowski3 |
| Abstract | The inheritance of the dehiscent pod character was investigated in two recombinant inbred populations using a simplified correlation analysis. The approach identified three regions on the pea genome that affect the expression of pod dehiscence. The region on linkage group III corresponded to the expected position of Dpo, a gene known to influence pod dehiscence. A locus on linkage group V appeared to have a slightly smaller effect on expression of the phenotype. The third region was observed only in one cross, had a greater effect than Dpo, and was postulated to be yellow pod allele at the Gp locus. |
| Address and Contact Information | 1Department of Plant Sciences and Plant Pathology, Montana State University,
Bozeman, MT, USA, 2Department of Biology, Ashland University, Ashland, OH, USA, 3Department of Plant Breeding and Seed Production, University of Warmia & Mazury in Olsztyn, Plac Łódzki 3, 10724, Olsztyn-Kortowo, Poland |
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| Volume 7 (2002) pp 665-670 | |
| Title | PHYSICAL MAPPING OF 18S-25S rDNA AND 5S rDNA IN LUPINUS VIA FLUORESCENT IN SITU HYBRIDIZATION |
| Authors | Barbara Naganowska and Anna Zielińska |
| Abstract | Double-target fluorescent in situ hybridization (FISH) was used to determine the genomic distribution of ribosomal RNA genes in five Lupinus species: L. cosentinii (2n=32), L. pilosus (2n=42), L. angustifolius (2n=40), L. luteus (2n=52) and L. mutabilis (2n=48). 18S-25S rDNA and 5S rDNA were used as probes. Some interspecific variation was observed in the number and size of the 18S-25S rDNA loci. All the studied species had one chromosome pair carrying 5S rDNA. |
| Address and Contact Information | Institute of Plant Genetics, Polish Academy of Sciences, Strzeszyńska 34, 60-479 Poznań, Poland |
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| Volume 7 (2002) pp 671-676 | |
| Title | THE USE OF RAPD AND SEMI-RANDOM MARKERS TO VERIFY SOMATIC HYBRIDS BETWEEN DIPLOID LINES OF Solanum tuberosum L. |
| Authors | Jarosław Przetakiewicz, Anna Nadolska-Orczyk and Wacław Orczyk* |
| Abstract | Markers specific to diploid lines of cultivated potato were identified using the polymerase chain reaction (PCR), primed with arbitrary 10-mers generating RAPDs, or with semi-random primers targeting intron-exon semiconservative sequences of plant genes. One hundred and fifty RAPD primers and twelve semi-random primers were tested. Selected primers were subsequently used for verification of putative somatic hybrids. |
| Address and Contact Information | Plant Breeding and Acclimatization Institute, Radzików, 05-870 Błonie, Poland * Corresponding author, E-mail: w.orczyk@ihar.edu.pl |
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| Volume 7 (2002) pp 677-684 | |
| Title | THE USE OF AFLP MARKERS IN CONSERVATION GENETICS - A CASE STUDY ON PULSATILLA VERNALIS IN THE POLISH LOWLANDS |
| Authors | Michał Ronikier* |
| Abstract | Pulsatilla vernalis is a rare species in the Polish lowlands, strongly threatened by anthropogenic disturbance of its habitats. A grave decrease in its populations has been observed during the past 60-80 years (analogous populations in Eastern Austria and the Czech Republic are almost or completely extinct). An analysis of the genetic diversity of populations in the Polish lowlands was performed to estimate its level and distribution. The AFLP method was used for the study of seven populations. An analysis using five pairs of selective primers revealed 446 scorable fragments; 62.1% of them were polymorphic. The average gene diversity indices was 0.17 (the mean value for all the populations), ranging from 0.139 to 0.204. A weak relationship between diversity and population size was revealed. Most of the genetic diversity was contributed to by the within-population level (AMOVA) and only a weak geographical structure was shown by UPGMA clustering. Four populations formed population-specific clusters while three others (from one region) were intermixed. These preliminary results show a moderate genetic diversity of the studied populations, which was still rather high when compared with their size. This result, together with the low between-population differentiation in the region, suggests that these populations are the remnants of larger populations that, only a few decades ago, were much less isolated. |
| Address and Contact Information | W. Szafer Institute of Botany, Polish Academy of Sciences, Lubicz 46,
31-512 Kraków, Poland; Laboratoire debotanique évolutive, Institut de
botanique, Université de Neuchâtel, Emile-Argand 11, 2007Neuchâtel,
Switzerland * Corresponding author, E-mail: ronikier@ib-pan.krakow.pl |
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| Volume 7 (2002) pp 685-694 | |
| Title | GENETIC DIVERSITY OF THE NOVI SAD WHEAT CORE COLLECTION REVEALED BY MICROSATELLITES |
| Authors | Borislav Kobiljski1, Steve Quarrie2* , Srbislav Denčić1, Jane Kirby1 and Mirjana Ivegeš2 |
| Abstract | In recent years, considerable emphasis has been placed on the development of microsatellites to be used for a variety of objectives. Parental genetic diversity is a crucial requisition to derive desirable and superior progenies from crossing and selection. In order to determine desirable genotypes for hybridization, 710 wheat genotypes from the Novi Sad Core Collection, originating from 38 countries, have been evaluated during the 1993-2000 period. During those seven growth seasons, 54 agronomical, morphological, physiological and other traits have been evaluated in field and controlled conditions. In each year, the field experiment comprised 3-7 replications, while for each field replication the plot size was 1.2 m2. Based on the results from this evaluation, 96 genotypes with the highest phenotypic variation for 26 of the very important traits for wheat breeding programmes in Yugoslavia and the UK, were identified for screening with microsatellites. A set of 36 microsatellite markers was used, covering all three wheat genomes and all 42 chromosomes. For the 36 microsatellites, a total of 46 loci and 366 alleles were detected, with the average number of 7.96 alleles per locus. For 35 loci, null alleles were detected. The association of microsatellite data with phenotypic data, for 6 important traits for wheat breeding (stem height, earliness, resistance to leaf rust and powdery mildew, sedimentation value and protein content), as well as the potential for their implementation in marker assisted selection (MAS) in wheat breeding programmes for both Yugoslavia and UK are discussed. |
| Address and Contact Information | 1Institute of Field and Vegetable Crops, M.Gorkog 30, 21000 Novi Sad,
Yugoslavia, 2John Innes Centre, Norwich Research Park, Colney, Norwich NR4 7UH, UK * Present address: Faculty of Agriculture, University of Belgrade, Nemanjina 6, 11080 Belgrade-Zemun, Yugoslavia |
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| Volume 7 (2002) pp 695-702 | |
| Title | THE IMPACT OF MOLECULAR MARKERS ON THE WHEAT BREEDING PARADIGM |
| Authors | Robert Koebner1 and Richard Summers2* |
| Abstract | We briefly review the limited application of marker assisted selection in past wheat breeding programmes, and contrast the current situation, where increasingly it has become feasible to tag almost any gene with a microsatellite assay. Although this capability is having an impact on the conduct of large breeding programmes, a much more profound change in breeding strategy will become possible when SNP technology has matured sufficiently so that the throughput of molecular marker-based genotyping will be able to keep pace with the numbers of plants that breeders can handle in the field. We discuss the considerations that will need to be addressed in the generation of a new breeding paradigm to take advantage of the genomics revolution. |
| Address and Contact Information | 1John Innes Centre, Norwich Research Park, Colney Lane, Norwich NR4 7UH,
UK, 2Monsanto UK Ltd., The Maris Centre, 45 Hauxton Road, Trumpington, Cambridge CB2 2LQ, UK * Corresponding author |
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| Volume 7 (2002) pp 703-708 | |
| Title | IDENTIFICATION OF ZYGOTIC AND NUCELLAR SEEDLINGS IN CITRUS INTERPLOID CROSSES BY MEANS OF ISOZYMES, FLOW CYTOMETRY AND ISSR-PCR |
| Authors | Nicasio Tusa1*, Loredana Abbate1, Sergio Ferrante1, Sergio Lucretti2 and Maria-Teresa Scarano1 |
| Abstract | 'Milam' (a purported hybrid of Citrus jambhiri Lush) + 'Femminello' lemon (Citrus limon L. Burm. f.) allotetraploid somatic hybrids were used as pollen parents in interploid crosses with diploid "Femminello" lemon to achieve mal secco tolerance in different populations of seedless triploid lemon types with good fruit quality. A total of 137 plantlets were obtained and subjected to screening experiments, in order to distinguish zygotic embryos from nucellars. Here we report on and discuss the results obtained with three techniques: flow cytometry, isozyme analysis and ISSR-PCR (the inter-simple sequence repeatspolymerase chain reaction). ISSR-PCR resulted to be a very efficient and reliable technique for the identification of zygotic plantlets. |
| Address and Contact Information | 1Istituto di Ricerca per la Genetica degli Agrumi - C.N.R. Viale delle Scienze n.
11, 90128-Palermo, Italy, 2ENEA C.R. Casaccia, Sezione Genetica e Genomica Vegetale (026) Via Anguillarese, 301-00060 S.M. di Galeria (Roma), Italy * Corresponding author, E-mail: ntusa@unipa.it |
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| Volume 7 (2002) pp 709-719 | |
| Title | REPRODUCIBLE TRANSFORMATION IN TWO GRAIN LEGUMES - SOYBEAN AND AZUKI BEAN - USING DIFFERENT SYSTEMS |
| Authors | Hany A. El-Shemy1, Mutasim Khalafalla1, Kyo Wakasa2,3 and Masao Ishimoto1,3 |
| Abstract | Two plasmid vectors were introduced into soybean (Glycine max (L.) Merr.) and azuki bean (Vigna angularis Willd. Ohwi & Ohashi) using different transformation systems. Azuki bean epicotyl explants were prepared from etiolated seedlings and co-cultivated with Agrobacterium tumefaciens for 2 days. Adventitious shoots were developed from the callus of the explants on a regeneration medium containing hygromycin, and the shoots were excised and transferred to a rooting medium containing hygromycin at the same concentration. Rooting shoots were transferred to soil and grown in a glasshouse to produce viable seeds. PCR analysis confirmed clearly the presence of the hpt gene in most of the azuki beans regenerated under hygromycin selection. A soybean embryogenic suspension culture was generated from immature cotyledons, and used for the introduction of plasmids by particle bombardment. Hygromycin-resistant embryogenic clones were isolated after 8 weeks of hygromycin selection, and then the green clones were matured on the differentiation medium. After desiccation, the embryos were germinated on the rooting medium, and the plants were transferred to soil in a glass-house. More than 50% of the regenerated soybean plants tolerant to hygromycin yielded the hpt fragment on PCR analysis. The azuki bean transformants were obtained more rapidly and with higher efficiency than the soybean transformants. |
| Address and Contact Information | 1National Agricultural Research Center for Western Region, 6-12-1
Nishifukatsu, Fukuyama, Hiroshima, 721-8514 Japan, 2National Institute of Crop Science, 2-1-18 Kannondai, Tsukuba, Ibaraki, 305-8518 Japan, 3CREST of JST (Japan Science and Technology Corporation), 3-4-15 Nihonbashi, Chuouku, Tokyo 103-0027, Japan * Corresponding author: E-mail: ishimoto@affrc.go.jp |
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| Volume 7 (2002) pp 721 -736 | |
| Title | LINKAGE GROUPS AND THE INDIRECT CHROMOSOME LOCATION OF cms-P-LINKED AFLPs |
| Authors | Piotr T. Bednarek1, Renata Lewandowska1, Helena Kubicka1 and Piotr Masojć2 |
| Abstract | Twenty-five AFLPs, previously linked to fertility restoration genes for the male-sterilizing PAMPA cytoplasm (cms-P) in a restricted rye population, were studied in an enlarged population of 120 plants. A strong association with the trait was verified for 19 of the markers. The recombination of these markers was tested and two linkage groups were identified: one consisting of six and the other of eight AFLPs. The remaining markers were segregated as independent loci. Using wheat-rye addition and substitution lines, the AFLPs were assigned to individual rye chromosomes. AFLP profiles of such lines were generated to identify the DNA fragments co-migrating with individual markers. This identified 1R and 3R as the two chromosomes corresponding to the linkage groups of eight and six markers, respectively. Mapping in a DS2 x RXL10 population linked four additional AFLPs to chromosome arms 1RS, 3RL, 4RL and 6RL. RAPD and SSR markers mapped in various populations and known to be located on the appropriate chromosomes did not disrupt the C394-F2 population into sterile and fertile phenotypes. It was concluded that the identified markers would reduce by one half the number of primer pair combinations needed for molecular breeding programs or for the selection of parental forms for rye hybrid crosses. |
| Address and Contact Information | 1The Botanical Garden-the Centre for the Conservation of Biological Diversity
of the Polish Academy of Sciences, 02-973 Warsaw, ul. Prawdziwka 2, Poland,
2Agricultural University of Szczecin, 71-434 Szczecin, Słowackiego 17, Poland * Corresponding author, E-mail: tmol.ob@ihar.edu.pl |
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| Volume 7 (2002) pp 737-744 | |
| Title | MOLECULAR MARKERS: TOOLS TO IMPROVE GENEBANK EFFICIENCY |
| Authors | Theo J.L. Van Hintum and Rob Van Treuren |
| Abstract | Possibilities for using molecular markers to improve genebank efficiency are increasingly present thanks to developments in genebanks and developments in molecular genetics. These possibilities relate to all aspects of genebank management: acquisition, maintenance, characterisation and utilisation. However, two pitfalls should be avoided. The first lies in the neutrality of the most generally used markers, making them less suitable for optimising genetic diversity. The second is related to the considerable costs involved in using molecular markers. In many cases an economical analysis will have to decide if the markers can routinely be used in genebank operations. Some examples of model studies and applications of molecular markers in genebank operations will be presented, in which both genetic and economic aspects will be illustrated briefly. These examples involved existing genebank collections of wild lettuce, cabbage and wild potato. |
| Address and Contact Information | Centre for Genetic Resources The Netherlands (CGN), Plant Research International, P.O. Box 16, 6700 AA Wageningen, The Netherlands |
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| Volume 7 (2002) pp 745-751 | |
| Title | MICROSATELLITE MARKERS DISCRIMINATING ACCESSIONS WITHIN COLLECTIONS OF PLANT GENETIC RESOURCES |
| Authors | Ján Kraic1, Edita Gregová1, Klaudia Jomová2 and Martina Hudcovicová1 |
| Abstract | The reliability of microsatellite analyses for discriminating between plant accessions maintained in collections of genetic resources was tested for 53 accessions of barley, 65 of soybean, 49 of chickpea, and 19 of alfalfa. The specific primer pairs used in this study were based on microsatellite DNA sequences surrounded by perfect dinucleotide and imperfect trinucleotide tandem repeat units. The evaluated polymorphic information content, diversity index, and probabilities of identity indicate that there is value in the application of SSR analyses in barley, soybean, and chickpea genetic resource management. Variation between alfalfa genotypes was not revealed at the five analyzed microsatellite loci. |
| Address and Contact Information | 1Research Institute of Plant Production, Bratislavská cesta 122, 92168 Piešťany,
Slovakia, 2Constantine the Philosopher University, Tr. A. Hlinku 1, 94901 Nitra, Slovakia |
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| Volume 7 (2002) pp 753-762 | |
| Title | MOLECULAR RESEARCH ON THE GENETIC DIVERSITY OF POLISH VARIETIES AND LANDRACES OF PHASEOLUS COCCINEUS L. AND PHASEOLUS VULGARIS L. USING THE RAPD AND AFLP METHODS |
| Authors | Jarosław Nowosielski, Wiesław Podyma and Dorota Nowosielska |
| Abstract | The aim of our research was to evaluate the genetic diversity among 25 commercial varieties registered in Poland and 14 landraces of Phaseolus vulgaris var. nanus Asch. (the dwarf common bean) and Phaseolus coccineus L. (the runner bean) maintained in the National Centre of Plant Genetic Resources in Radzików. An additional goal of this study was to compare the precision and efficiency of two techniques of PCR (RAPD and AFLP), used to estimation the genetic diversity of bean. The breeding varieties of bean were registered in the period between 1950 and 2000. The landraces, collected during expeditions conducted from 1985 to 1988, mainly originated from the eastern and southern part of Poland. In the plant genetic diversity research of RAPD and AFLP markers are commonly used. Complex electrophoresis pictures of DNA fragments were taken, and revealed a considerable polymorphism. The polymorphic fragments were obtained on the basis of 6 differentiating primers using the RAPD method and 15 differentiating primers using the AFLP method. P. vulgaris and P. coccineus accessions formed distinct groups. Each of the RAPD and AFLP analyses allowed for the unique distinguishing of all accessions. |
| Address and Contact Information | National Centre for Plant Genetic Resources, Plant Breeding and Acclimatization Institute, Radzików, 05-870 Blonie, Poland |
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| Volume 7 (2002) pp 763-769 | |
| Title | GENETIC MAPPING OF POLYPHENOL OXIDASE IN TETRAPLOID WHEAT |
| Authors | Rosanna Simeone1*, Antonella Pasqualone2, Maria Lisa Clodoveo1 and Antonio Blanco1 |
| Abstract | Pasta colour is one of the main factors influencing pasta quality. It is the product of a desirable yellow component, an undesirable brown component and, under some drying conditions, a red component. The brown colour depends on enzymatic and chemical factors. Polyphenol oxidase (PPO; E.C. 1.14.18.1) is one of the enzymatic factors. It is mainly localised in the peripheral part of the wheat kernel, and is involved in the oxidation of endogenous wheat phenolic compounds resulting in the production of highly coloured products. Therefore, a knowledge of the genetic control of PPO activity could enable the developing of better strategies in breeding programs to reduce pasta darkening. The aim of this study was to map the gene(s) affecting PPO activity using a set of recombinant inbred (RI) lines, derived from a cross between Triticum turgidum L. var. durum cultivar Messapia and the accession MG4343 of Triticum turgidum L. var. dicoccoides. After performing linkage analysis, the gene for high PPO activity was mapped on the long arm of the chromosome 2A and its characteristic was found highly associated to the RFLP marker Xutv1427-2A, with a value of LOD equal to 29.84. The identification of molecular markers linked to loci controlling the PPO activity may potentially accelerate wheat breeding since the selection of plants can be carried out by genotype rather than phenotype. |
| Address and Contact Information | Institute of Plant Genetics and Crop Science, Department. of Taxonomy, Corrensstrasse 3, D-06466 Gatersleben, Germany |
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| Volume 7 (2002) pp 771-776 | |
| Title | TAGGING QTLS FOR MAXIMUM ROOT LENGTH IN RAINFED LOWLAND RICE (ORYZA SATIVA L.) USING MOLECULAR MARKERS |
| Authors | Mahmoud Toorchi1*, H. E. Shashidhar2, Naveen Sharma and Shailaja Hittalmani2 |
| Abstract | A number of morphological, physiological and phenological traits are known to improve the performance of rice challenged by drought. Root morphological traits and stress-induced response form important components of drought tolerance. Enhancing grain yield remains the principal objective of most breeding programs. Interaction between primary traits poses a formidable challenge while dealing with grain yield under stress. The evaluation of root morphology at three different growth stages and grain yield along with related characteristics under contrasting moisture regimes was made using nine backcrosses along with their parent and standard checks. The backcrosses invoved transgressant double haploid lines derived from IR64 and Azucena with IR64. Marked genotypic differences were observed for all root morphology as well as grain yield related characteristics across the sampling dates as revealed by individual and combined ANOVA. Among the nine backcrosses studied in this experiment, the BC1F2 population of P124 x IR64 were evaluated for forwarding based on their performance with respect to maximum root length and grain yield under both well-watered and low-moisture stress conditions. Sixtynine plants-ten percent of the backcross population-were selectively genotyped using RAPD primers. Under well-watered conditions two RAPD markers showed strong linkage to QTLs for maximum root length evaluated under ww conditions. Two other markers could explain the considerable amount of variation in MRL under LMS. One of the markers identified under lowmoisture stress conditions was also able to explain variability in maximum root length in the mean environment. |
| Address and Contact Information | 1Department of Agronomy, Faculty of Agriculture, Tabriz University, Tabriz,
Iran, 2Marker Assisted Selection Lab., University of Agricultural Sciences, G.K.V.K., Bangalore 560065, India * Corresponding author, Email: mtoorchi@yahoo.com, Fax +(98-411) 3345332 |
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| Volume 7 (2002) pp 777-783 | |
| Title | THE APPLICATION OF THE AFLP METHOD TO DETERMINE THE PURITY OF HOMOZYGOUS LINES OF BARLEY (HOREDUM VULGARE L.) |
| Authors | Sylwia Oleszczuk1, Janusz Zimny1 and Piotr Tomasz Bednarek2 |
| Abstract | Amplified fragment length polymorphism of DNA has been used to analyse the equality of plants obtained from isolated microspores. Although the control parental material was regarded as being highly homozygous, the analysis of the banding patterns of single plants showed a certain level of polymorphism. The analysis of regenerants with a doubled chromosome number did not show any diversity within the progeny of a single line. The differences in banding patterns coming from single plants were only observed in microspore donor lines. These results have proven the high purity of homozygous lines obtained via androgenesis from isolated microspores. |
| Address and Contact Information | 1Department of Biotechnology and Plant Cytogenetics, Plant Breeding and
Acclimatization Institute, Radzików, 05-870 Błonie, Poland, 2The Botanical Garden-the Centre for the Conservation of Biological Diversity of the Polish Academy of Sciences,Prawdziwka 2, 02-973 Warsaw, Poland |
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| Volume 7 (2002) pp 785-794 | |
| Title | FRUIT PLANT GERMPLASM CHARACTERISATION USING MOLECULAR MARKERS GENERATED IN RAPD AND ISSR-PCR |
| Authors | Małgorzata Korbin, Anita Kuras and Edward Żurawicz |
| Abstract | The genotypes of the strawberry (Fragaria x ananassa), apple (Malus domestica) and Ribes species (R. nigrum, R. rubrum and R. glossularia), maintained in our Institute's collection and used in breeding programs, were screened for DNA markers. Twenty primers for RAPD (among 60 tested) and seven for ISSR (among 10 tested) were chosen as creating polymorphic DNA bands differentiating the investigated genotypes. Based on those identity markers, the genetic distance between genotypes was determined, and their relatedness was estimated. In many cases, both RAPD- and ISSR-based genetic similarity confirmed relatedness connected with biological origin and with the place where the cultivar was developed. However, some diversity connected with the technique used for molecular marker generation was observed. Generally, the similarity values based on ISSR data were higher than those based on RAPD. Parallel study using two data sets seems to enable a reduction in the number of potential mistakes connected with each method's, technical limitations and ensures more precise relatedness determination. |
| Address and Contact Information | Research Institute of Pomology and Floriculture, Pomologiczna 18, 96-100 Skierniewice, Poland |
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| Volume 7 (2002) pp 795-802 | |
| Title | GENETIC MAPPING AND TAGGING OF WHEAT GENES USING RAPD, STS AND SSR MARKERS |
| Authors | Elena K. Khlestkina1, Elena G. Pestsova1, Elena Salina1, Marion S. Röder2, Valentina S. Arbuzova1, Sergej F. Koval1 and Andreas Börner2 |
| Abstract | We applied SSR markers for mapping genes determining red coleoptile colour in wheat (Rc1, Rc2, Rc3) using F2 populations. All three genes map at about 15 to 20 cM distally from the centromere of chromosomes 7AS, 7BS and 7DS, respectively. The locations of the glume colour (Bg, Rg1) and glume hairiness (Hg) genes relative to the SSR markers of the homoeologous chromosomes group 1 were determined using molecular analysis of nearisogenic lines (NILs). One RAPD marker for the vernalisation response gene Vrn-A1 was identified by screening 95 random primers against two pairs of NILs. New PCR (STS) markers were developed based on RFLP-markers PSR426 (5A, 5B, 5D) and PSR1201 (1A, 5A, 5B). Analysis of nulli-tetrasomic and near-isogenic lines of wheat using the STS markers developed gave an indication that these new STS markers have the same chromosomal and intrachromosomal positions as the correspondent RFLP markers. Therefore, they could be used for mapping and/or tagging the vernalisation response (Vrn-A1, Vrn-B1, Vrn-D1) and homoeologous pairing (Ph1) genes. |
| Address and Contact Information | 1Institute of Cytology and Genetics, Siberian Branch of the Russian Academy of
Sciences, Novosibirsk, 630090, Russia, 2Institut fur Pflanzengenetik und Kulturpflanzenforschung (IPK), Corrensstr. 3, D-06466 Gatersleben, Germany * Corresponding author, khlest@bionet.nsc.ru |
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| Volume 7 (2002) pp 803-810 | |
| Title | RAPID GENETIC MAPPING OF ESTs USING SNP PYROSEQUENCING AND INDEL ANALYSIS |
| Authors | Ada Ching* and Antoni Rafalski |
| Abstract | We describe an effective systematic approach to genetic mapping of cDNA clones, including those obtained from EST sequencing. The EST of interest is first partially sequenced from the 3'-end. PCR primers which bracket the 3'-UTR segment of the cDNA are designed. The corresponding gene segment is amplified from the parents of the mapping population, using primers equipped with 3'- and 5'-extensions to facilitate direct sequencing of PCR products. Comparison of the sequences obtained from the mapping parents frequently reveals single nucleotide polymorphisms or insertion / deletion polymorphisms, which can then be genotyped in a mapping population. The genotyping of SNPs is performed by pyrosequencing, a sequencing-by-synthesis method that has been used successfully in SNP diagnostics. SNP analysis of up to 96 samples, a number required to produce meaningful genetic segregation data, can be rapidly accomplished in parallel. The parental genotype of three loci, stearoyl-ACP desaturase, nucleoside-diphosphate kinase and sucrose synthetase-1 were determined by conventional sequencing, and the polymorphism so identified were scored by the pyrosequencing of 94 individuals of a maize recombinant-inbred population. These loci were successfully placed onto chromosomes 3, 7 and 9 respectively. This method is generally applicable to most plant species, which show sufficient sequence diversity in the 3'-UTR region of genes. |
| Address and Contact Information | DuPont Crop Genetics, Molecular Genetics Group, 1, Innovation Way,
Newark, DE 19711, USA * E-mail: Ada.S.Ching@USA.dupont.com |
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| Volume 7 (2002) pp 811-820 | |
| Title | USE OF MOLECULAR MARKERS IN CEREAL BREEDING |
| Authors | Viktor Korzun* |
| Abstract | Great advances have been made in recent years in marker detection systems and in the techniques used to identify markers linked to useful traits. While RFLP markers have been the basis for most work in crop plants, useful markers have been generated using RAPD and AFLP methods. More recently, microsatellite or simple sequence repeat (SSR) markers have been developed for major crop plants and this marker system is predicted to lead to even more rapid advances in both marker development and implementation in breeding programs. Identification of markers linked to useful traits has been based on complete linkage maps and bulked segregant analysis. However, alternative methods, such as the construction of partial maps and combination of pedigree and marker information, have also proved useful in identifying marker/trait associations. The value of markers in analysing the inheritance of traits in crop plants and understanding genome structure and organization is now well established. The different properties of markers systems and their applications in genome analysis and molecular breeding of cereals species are discussed. |
| Address and Contact Information | Lochow-Petkus GmbH, PF 1197, D-29296 Bergen, Germany |
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