ISSR-PCR Technique Research Articles

Molecular and biochemical markers in some lentil (Lens culinaris Medik.) genotypes

Lentil (Lens culinaris Medik.) is a major winter-sown legume crop grown in the Mediterranean region, and has considerable importance as food and fodder. The SDS–PAGE and ISSR-PCR techniques were used to detect some biochemical and molecular markers associated with drought tolerance in six local and exotic genotypes of lentil (Sinai 1, Giza 9, L 9, L 16, L 20 and L 25) under rainfed conditions at El-sheikh Zowaid, North Sinai. Sinai 1 and L 25 gave the highest level of yielding capacity and could be considered as drought tolerant genotypes, while L 20 and L 9 as moderately sensitive ones. Seed proteins gave low levels of genetic diversity. Five ISSR primers were used and six bands considered as molecular markers for drought tolerant genotypes.

Production and characterization of new triploid seedless progenies for mandarin improvement

Eighty-six triploid Citrus plants were recovered from interploid crosses between a natural tetraploid selection of the tangerine ‘Dancy’ ( Citrus reticulata Blanco, cultigroup ‘Tangerine’), used as the pollen parent, and two seedy selections of diploid mandarins ( C. reticulata Blanco) as well as one clementine ( Citrus clementina Hort. ex Tan.): ‘Fortune’ mandarin, ‘Wilking’ mandarin and ‘Monreal’ clementine. Flow cytometric analysis was used for screening the triploid plantlets and the ISSR-PCR technique was used to characterize the obtained triploids through a double approach, confirming the hybrid nature of the offspring and allowing the analysis of the genetic pool obtained. Selection among triploid genotypes generated from elite seedy parents will give us a better chance to obtain superior mandarin cultivars characterized by true seedlessness.

Genetic diversity and genetic differentiation of natural Pinus koraiensis population

Genetic diversities and genetic differentiations of the four Pinus koraiensis populations (Gaofeng Forestry Farm at Tangwanghe in Yichun City, Erdaobaihe Town in the Changbai Mountains, Shengshan Forestry Farm in Heihe City, in China, and the suburb of Vladivostok City in Russia) were analyzed by using ISSR-PCR technique. The results of 15 primers amplification showed that the ratio of polymorphic site of P. koraiensis population was 60.7%. Each primer had 3.6 polymorphic sites. The diversity levels of the four P. koraiensis populations were rather higher compared with those of other Pinaceae species. The genetic diversity levels of P. koraiensis populations grown in the center region were higher than those grown in the edge zone. The genetic diversity of P. koraiensis mainly came from the interior of the population, accounting for 73% of total genetic diversity. There were no positive correlation between genetic distances and geographical distances for the four P. koraiensis populations. The gradual decrease of natural distribution region of P. koraiensis was due to anthropic destroy and environmental factors (i.e. fire and wind throw), rather than the lower genetic diversity.

DNA fingerprint of F1 interspecific hybrids from the Triticeae tribe using ISSRs

Inter-simple-sequence repeat (ISSR) were used to fingerprint three kinds of F1 interspecific hybrids: the Portuguese durum wheat cultivar ‘Candial’ × tritordeum, the Portuguese bread wheat cultivar ‘Barbela’ × tritordeum and the Portuguese triticale cultivar ‘Douro’ × tritordeum. Among the 30 primers tested, the number of discriminative primers varies from 13 to 20 in each hybrid combination, the total number of ISSR loci analysed ranged from 101 to 183, and the number of discriminative ISSR loci varied from 50 to 71. ISSR-PCR technique revealed to be a valuable fingerprint system for all F1 hybrid combinations and respective parents analysed here. The complementary use of molecular cytogenetic techniques and molecular markers could allow a more accurately evaluation and characterisation of different plant species.

Diversity and molecular relationships of endophytic Colletotrichum isolates from the Iwokrama Forest Reserve, Guyana

The diversity and host specificity were studied of a collection of Colletotrichum strains derived from endophytic colonies in leaves of 12 tree species in the Iwokrama Forest Reserve, Guyana. Analysis included ISSR-PCR and RAPD molecular fingerprinting techniques, rDNA ITS sequencing and morphological and cultural characterization. Most strains belonged to one of two species, C. gloeosporioides and a further taxon which is probably referable to C. boninense. Almost no strains were found to be genetically identical, indicating that clonal reproduction does not play a prominent role. No degree of host specificity could be detected even at molecular fingerprint level. The implications for estimation of fungal diversity in closed tropical forests may be profound.

Assessment of Genetic Polymorphism in Hop (Humulus lupulusL.) Cultivars by ISSR–PCR Analysis

Genetic diversity among 26 Russian and European cultivars of the common hop (Humulus lupulusL.) was studied using the ISSR–PCR technique. Twenty-one primers used provided amplification of 183 DNA fragments, 106 of which (57.9%) were found to be polymorphic. The ISSR markers, specific for some cultivars were revealed. Based on the genetic distances, cluster analysis was performed and a dendrogram was constructed, on which most of the hop cultivars formed two clusters according to their origin. Advantages of the ISSR–PCR analysis in breeding studies for classification and identification of common hop cultivars are discussed.

Temporal regulation of behaviour in a fresh water turtle, [formula omitted] (Wied)

The yellow-necked field mouse, Apodemus flavicollis (Melchior, 1934), and the long-tailed field mouse, Apodemus sylvaticus (Linnaeus, 1758) are morphologically similar species, with largely overlapping geographical areas and almost equal ecological requirements. This makes reliable species diagnosis based on external characters a real challenge. When advanced multivariate methods of skull and/or dental morphometrics are employed, specimens can be successfully distinguished in most cases, but not all, making single specimen identification impossible. Application of C-band karyotyping clearly distinguishes between these two species. However, it can be applied only to live animals. Several molecular methods have also allowed successful species diagnosis, but their universality has not been tested. Here we compare the diagnostic power of three molecular approaches and their applicability to populations from a species wide distributional area, as well as their simplicity of use, both for live animals and for samples stored in alcohol for a considerable period, without DNA sequencing. A total of 200 tissue samples from Morocco, France, Belgium, Serbia, Romania, Greece, Russia, Turkey and Kazakhstan, were analyzed by AP-PCR, ISSR-PCR and species-specific primers from the mitochondrial cytochrome b gene. The first two methods gave clear species-specific DNA profiles in complete agreement with previous C-band results. However, only partial agreement was observed between the third, species-specific primer method and the other two approaches and C-banding, as there were 4.5–10.5% false positive results. Hence we propose AP-PCR and ISSR-PCR techniques with particular chosen primers for quick and reliable diagnosis of these two species. A large number of samples can be assayed in a short period, with minimal cost and effort. Moreover, these techniques can be applied to a single specimen.