Sweet cherry cultivar identification by using SSR markers

In this study, sweet cherry varieties and types grown in Turkey were described using SSR markers. This study was undertaken to develop DNA marker profiles that could be used to distinguish among the sweet cherry cultivars used in production in Turkey. For microsatellite analysis, 13SSR primers isolated from sweet cherry (P. avium L.) sour cherry (P. cerasus L.) and peach (P. persica L. Batsch) were used on sweet cherry cultivars and types. Two primer pairs did not give amplification with genotypes analyzed. Two-primer pairs amplified monomorphic fragment for the sweet cherry varieties therefore they were uninformative for the sweet cherry genetic analysis. Genetic similarities were calculated and a dendrogram has been established. All of the 8 SSR primers used and cherries have produced amplified bands. For each primer the alleles obtained has been between 1 and 6, in total 38 alleles have found. Through these analyses the similarities between these varieties have been converted into numerical values. This happens to be the first study kind towards the molecular identification of sweet cherry genetic resources in Turke

Assessment of genetic variability between inbred and siblines of latvian origin cucumber population using RAPD markers

The proper management of plant genetic resources, ex situ conservation in gene banks requires the selection of genetically distinct accessions for long-term preservation. The conservation of vegetable resources in the Latvian Gene Bank is prioritised particularly for the cultivars of local origin, old cultivars and landraces. Due to improper seed propagation the old cucumber cultivar 'Dindona Zalie Kekaru' was contaminated and became a population with a broad spectrum of phenotypic and genotypic diversity. Population dividing in different genotypes was started by inbreeding and family selection (sibline). This study aimed to detect genetic diversity between lines by using RAPD markers. In total 53 primers were tested for 17 inbred and siblines, derived from the population of 'Dindona Zalie Kekaru', and for one sample of a local cultivar 'Grïvas'. We find polymorphism between lines for 26 primers, one primer was monomorphic, and with the other 26 primers there was no stated amplification. As expected, middle to high genetic diversity was established between the lines. Polymorphic loci varied from 50 to 100%

In vitro propagation of Ficus carica L. var. bursa siyahi through meristem culture

1st International Symposium on Fig -- JUN 24-28, 1997 -- IZMIR, TURKEYWOS: 000079157300028Bursa Siyahi is a well-adapted fig cultivar under Cukurova conditions. However, almost all trees are infected with the fig mosaic virus. In vitro propagation of this cultivar through meristem culture was carried out in order to obtain virus-free plant propagation. The meristems were isolated during the growing seasons and cultured on Linsmair and Skoog (1965) medium, supplemented with 0.5 mg/l Benzyladenine (BA), 0.1 mg/l Indole Butyric Acid (IBA), 0.1 mg/l Gibberellic Acid (GA(3)) + 89 mg/l Phloroglucinol (PG) and + 2 g/l active charcoal (AC). Then, growing shoots were transferred to a shoot proliferation medium including 89 mg/l PG, 0.5 and 1.0 mg/l BAP. In order to induce rooting, propagated shoots were subcultured onto a rooting medium with 0, 1 and 2 mg/ 1 IBA. In the meristem phase, survival and shoot formation rate of meristems was investigated, in the proliferation phase, the rate of proliferation and the effect of subculture number on the proliferation rate and in the rooting phase, the rate of rooted explants and the number of roots per explant. At the end of the meristem phase, the meristems which were isolated from shoot tips taken in spring or autumn times and cultured on a medium with phloroglucinol or active charcoal showed the highest shoot formation rate (50.1 %). The propagation rate was found to be higher (4.43 plantlet/plant) in the medium containing 1.0 mg/l BAP than 0.5 mg/l BAP (3.52). At the end of the rooting experiments, differences between the auxin treatments were not found to be statistically significant; however, the highest rooting (75.0 %) was obtained in medium without auxin. This was followed by 1 mg/l and 2 mg/l IBA containing media having 68.33 and 55.33 % rooting, respectively.Int Soc Hort Sc

Clonal propagation of Pistacia rootstocks by meristem and shoot culture

The clonal propagation of rootstocks used for pistachio species such as Pistacia khinjuk Stock, Pistacia atlantica Desf., Pistacia terebintus L. and Pistacia integerrima Stewart via tissue culture techniques was investigated. Meristems and the shoots which were obtained by germinating the seeds were used as propagation materials. Different concentrations of BAP were used as cytokinin. Healthy shoot development was not obtained in Pistacia integerrima Stewart. In the other three species, shoot development was obtained in Murashige & Skoog medium containing 4 mg/1 BAP, but the problems were not solved completely. Meristems of Pistacia terebintus L., Pistacia khinjuk Stock, andPistacia atlantica Desf. showed 3.88, 2.20 and 2.20 multiplication ratio, respectively. The shoots of Pistacia terebintus L., Pistacia khinjuk Stock, Pistacia atlantica Desf. showed 1.97, 1.36 and 0.80 multiplication ratio, respectively. During in vitro propagation of Pistacia species, some problems, such as browning, death of shoot tips, vitrification, variation and callus production, were seen. In order to solve the problems which were encountered, some studies, such as subculturing frequently, usage of active charcoal, ascorbic acid, and citric acid, and changing media compositions were carried out. These problems were not eliminated totally, however

Morphological and genetic characterization of indeterminate green bean genotypes having different seed coat color collected from Turkey

The aim of this study was to determine some morphological characters and genetic relations of indeterminate green bean accessions having different seed coat color collected from the Black Sea region of Turkey. In this research, a total of 39 green bean accessions and one kidney bean accession consisting of different main seed coat colors (white, black, yellow, green, brown, red, and dark red) and mixed seed coat colors were used. Plants were produced under greenhouse conditions for morphological characterization. We used 43 morphological characters described by UPOV for indeterminate green bean. In molecular analysis, the sequence-related amplified polymorphism (SRAP) marker system was used. A total of 21 primer combinations produced 138 bands. DNA band profiles were analyzed by NTSYS and a UPGMA dendrogram was generated. The total polymorphism rate was found to be 85.5%. According to research results, indeterminate green bean accessions showed a high level of morphological variation; however, narrow genetic diversity was detected based on molecular analysis

Determination by SSCP markers of the allelic diversity of candidate genes for tolerance to iron chlorosis in citrus germplasm

Iron chlorosis is one of the main abiotic constraints for Mediterranean citriculture and the development of marker assisted selection (MAS) for this trait would be a great aid for rootstock breeding. We performed SSCP (Single Stranded Conformation Polymorphism) analysis in order to discover allelic diversity of candidate genes for iron chlorosis tolerance in citrus. Two iron chlorosis tolerance candidate genes were selected from existing Citrus ESTs databases and Arabidopsis thaliana genome databases. Iron-Sulfur assembly protein (Fe-S=AT2G16710) and root iron transporter (IRT1=AT4G19690) candidate gene sequences were used to define primers in conserved regions. Six Citrus genotypes from the basic taxon of Citrus where used to identify polymorphic areas in the genes. Direct sequencing of amplified DNA fragments of candidate genes was performed and SNPs (singlenucleotide polymorphisms) and Indels where searched after sequence alignment. A total of 2215 bp DNA fragments were sequenced and 56 SNPs and 2 Indels were determined. New primers were defined, in conserved areas flanking polymorphic ones, for Single Strand Conformation Polymorphism (SSCP) analysis for further diversity and routine genotyping. Two pairs of primers were defined for each gene. SSCP-PCR analysis was performed with twenty-five Citrus genotypes. The neighbor joining method was used for cluster analysis. Poncirus trifoliata genotypes and their hybrids known to be sensitive to iron chlorosis clustered together and mandarins showed high diversity for both genes. Differences were found among sour orange genotypes known to have differential tolerance behavior to iron chlorosis

In-vitro multiplication of clonal apple rootstocks M-9, M-26 and MM-106 by meristem culture

Meristems of apple rootstocks M-9, M-26 and MM-106 were cultured on 1/2 strength MS medium containing 2.2 uM BA, 0.5 uM IBA, and 0.3 uM GA3. M-26 and MM-106 shoots were transferred on Lepoivre and MS media, respectively, supplemented with 4.4 uM BA, 5 uM IBA, 0.3 uM GA3 The rate of shoot proliferation per mother explant of M-26 and MM-106 were 6 and 3.4, respectively. The best rooting of MM-106 was achieved with the use of 4 uM IBA, 0.3 uM giberellic acid (GA3) and 1300 uM phloroglucinol (PG) in which 80 % rooting was obtained. During the proliferation phase, shoot tip necrosis and vitrified shoots were observed with MM-106. To overcome these , PG was used. Although vitrification was not observed, the shoot elongation M-26 was limited to around 1 cm. In M-9, insufficient shoot proliferation was obtained, thus experiments are underway to increase shoot production

Genetic characterization of heat tolerant tomato (Solanum lycopersicon) genotypes by SRAP and RAPD markers

We employed RAPD and sequence-related amplified polymorphism (SRAP) markers to evaluate polymorphisms in 15 tomato (Solanum lycopersicon) genotypes that were obtained from a tomato breeding program. Four local tomato genotypes selected from the Sanliurfa province (Southeastern Anatolia Region of Turkey), 10 heat-tolerant tomato genotypes, received from the Asian Vegetable Research and Development Center, and a sample of S. pimpinellifolium were genotyped with RAPD and SRAP markers. Eleven SRAP primer combinations were used and 66 bands were scored. The number of bands scored per primer combination ranged from three to 12, with a mean of six alleles per primer combination. All fragments scored for each primer combination were polymorphic. The percentage of polymorphic products ranged from 25 to 80%. The 15 tomato genotypes were screened for RAPD markers using 50 primers in a PCR-based DNA amplification procedure; 46 primers produced clear and good amplification. Ten of these 46 primers amplified monomorphic fragments in the tomato genotypes. A dendrogram was constructed by combining data from the RAPD and SRAP analyses. Similarity ratios of genotypes ranged from 0.87 to 0.99. The dendrogram was divided into two branches; the first main branch included only genotype CL 5915, and the second main branch included all the other genotypes

Meristem culture of two fig cultivars in Turkey

Introduction. Two Turkish fig cultivars, Alkuden and Bursa Siyahi, were propagated by meristem culture to eliminate the fig mosaic virus. The technique of dsRNA analysis was conducted on the in vitro propagated plants to test for virus-free status. Materials and methods. Four different Murashige and Skoog (MS) media complemented with different concentrations of growth hormones (GA3, BA and IBA) were compared to study the cultured meristems' survival rate, shoot formation and rooting. Short- and long-term thermotherapy treatments were also applied. Results. For survival rate, 0.2 mg GA 3·L-1 + 0.5 mg BA·L-1 gave the most favorable results; for shoot formation, it was the medium with 0.2 mg GA3·L-1 + 2.0 mg BA·L-1 which was the best, while rooting rate was the highest for meristems cultured on MS medium with only 0.1 mg GA3·L-1. Conclusion. Meristem culture, together with thermotherapy treatment, is recommended to obtain virus-free fig plant material. Although cultured plants seemed to be very healthy, dsRNA virus tests are recommended for sensitive evaluation of the sanitary status of the plants obtained. © 2007 Cirad/EDP Sciences. All rights reserved

The utility of GFP in genetic engineering of horticultural plants

Reporter proteins play a significant role in developing and optimizing transformation protocols for plant species and they show a unique activity to visualize gene expression and protein localization. Chemical based selection for plant transformation is associated with a number of problems that might be avoided through visual selection. In recent years, the reporter protein GFP (Green Fluorescent Protein) has become very effective and highly valuable for use in biotechnology, cell biology, and biochemistry. It is a unique tool for monitoring gene expression, protein localization, detection of the gene flows and protein dynamics in both prokaryotic and eukaryotic living cells because it requires neither exogenous substrates nor cofactors for its activity. We discussed the use of jellyfish green fluorescent protein (GFP) as a reporter in horticultural plants. © Verlag Eugen Ulmer GmbH & Co