The prevalence of sour orange rootstock in the southern and eastern part of the Mediterranean Basin is presently threatened by the spread of Citrus Tristeza Virus (CTV) and its main vector Toxoptera citricida, combined with abiotic constraints such as drought, salinity and alkalinity. The search for alternative CTV-resistant rootstocks that also withstand the other constraints is now considered an urgent priority for a sustainable citrus industry in the area. Complementary progenitors can be found in citrus germplasm to combine the desired traits, particularly between Poncirus and Citrus genera. The production of somatic hybrids allows cumulating all dominant traits irrespective of their heterozygosity level, and would appear to be an effective way to solve the rootstock challenge facing the Mediterranean citrus industry. This paper presents the results obtained during a regional collaborative effort between five countries, to develop new rootstocks by somatic hybridization. New embryogenic callus lines to be used for somatic hybridization have been created. Protoplast fusions have been performed at CIRAD and IVIA laboratories, focusing on intergeneric combinations. Analysis of ploidy level by flow cytometry and molecular markers confirmed the acquisition of new interesting tetraploid somatic hybrids for six combinations. Diploid cybrids with intergeneric (Citrus × Poncirus) nucleus and C. reticulata or C. aurantifolia mitochondria were also identified for four combinations. The agronomical performance of a pre-existing somatic hybrid between Poncirus trifoliata and Citrus reticulata was validated in calcareous soils in Morocco. Somatic hybridization is now integrated into the breeding programs of the five Mediterranean countries.
Genetic diversity of the Turkish watermelon genetic resources was evaluated using different Citrullus species, wild relatives, foreign landraces, open pollinated (OP) and commercial hybrid cultivars by RAPD markers. The germplasm was consisted of 303 accessions collected from various geographical regions. Twenty-two of 35 RAPD primers generated a total of 241 reproducible bands, 146 (60.6%) of which were polymorphic. Based on the RAPD data the genetic similarity coefficients were calculated and the dendrogram was constructed using UPGMA (Unweighted pair-group method with arithmetic average). Cluster analysis of the 303 accessions employing RAPD data resulted in a multi-branched dendrogram indicating that most of the Turkish accessions belonging to var. lanatus of Citrullus lanatus (Thunb.) Matsum et Nakai were grouped together. Accessions of different Citrullus species and Praecitrullus fistulosus (Stocks) Pangalo formed distant clusters from C. lanatus var. lanatus. Among 303 accessions, a subset of 56 accessions was selected representing different groups and a second dendrogram was constructed. The genetic similarity coefficients (GS) within the Turkish accessions were ranged from 0.76 to 1.00 with 0.94 average indicating that they are closely related. Taken together, our results indicated that low genetic variability exist among the watermelon genetic resources collected from Turkey contrary to their remarkable phenotypic diversity.
The genus Crataegus known as hawthorns, is the largest genus among the Maloideae, which comprises 265 species. Turkey is one of the genetic centers of Crataegus and there are more than 20 species found in Turkey. The fruits of Crataegus are used as food and have high flavonoid, vitamin C, glycoside, anthocyanidin, saponin, tannin, and antioxidant levels. In this study, we attempted to characterize 15 Crataegus accessions sampled from Hatay, located in Eastern Mediterranean region of Turkey. The accessions belonged to several species; C. aronia (L.) DC. var. aronia, C. aronia var. dentata Browicz, C. aronia var. minuta Browicz, C. monogyna Jacq. subsp. azarella (Griseb.) Franco, and C. orientalis Pall. ex M. Bieb. var. orientalis. Fruit characteristics of the accessions exhibited considerable variation. The multivariate, principle component and cluster analyses indicated that the accessions belonged to three groups: (1) C. aronia var. arona accessions;(2) C. aronia var. dentata accessions; and, (3) C. monogyna subsp. azarella and C. orientalis var. orientalis accessions. The principle component analysis results also revealed that the first three components explained 46, 21, and 14% of the variation, comprising a total of 81%. The fruit length and width, leaf area, and soluble solids contents were highly correlated characteristics for the first three components. The 19 RAPD primers generated a total of 107 bands, where 76 of these were polymorphic. The molecular data analyses by principle coordinate and clustering showed similar results to those of pomological characteristics. There were three groups, (1) C. aronia var. arona accessions;(2) C. aronia var. dentata accession; and, (3) C. monogyna subsp. azarella. C. orientalis var. orientalis accession grouped with C. aronia var. arona accessions. Therefore, it can be concluded that, overall, the diversity patterns of pomological and molecular data, generated by RAPD, for Crataegus are in good agreement and the accessions of C. aronia var. aronia, C. aronia var. minuta, C. monogyna subsp. azarella and C. orientalis var. orientalis accessions.
Grapefruit is the fourth economically most important citrus fruit in the world. In this research Inter-Simple Sequence Repeat (ISSR) markers were used to distinguish twenty-nine grapefruit (Citrus paradisi Macf.), five pummelo (Citrus maxima (Burm.) Merr.) and one Citrus hassaku Hort. Ex Tanaka accessions. Twelve ISSR primers produced a total of 100 fragments and 62 of them were polymorphic. The number of average polymorphic fragments per primer was 5.2. The mean polymorphism information content (PIC) was 0.37. The unweighted pair group method arithmetic average (UPGMA) analysis demonstrated that the accessions had a similarity range from 0.79 to 1.00. The accessions were separated into two main clusters; group A with five pummelos and group B with grapefruits. In the pummelo cluster, all pummelos were distinguished whereas in the grapefruit cluster some accessions were not clearly separated. There was a low level of variation in the grapefruits due to their mutation origin.
Genetic diversity was evaluated by sequencerelated amplified polymorphism (SRAP) and simple sequence repeat (SSR) markers among 45 lemons (Citrus limon (L.) Burm. f.), five citrons (Citrus medica L.), four rough lemons (Citrus jambhiri Lush), and two Citrus volkameriana accessions. Twenty-one SRAP primer combinations produced a total of 141 (77%) polymorphic fragments with an average of 6.7 fragments per primer combinations whereas 13 SSR primers produced a total of 26 (76%) polymorphic fragments with an average of 2.0 per primer. The unweighted pair-group method arithmetic average analysis as assessed with combined SRAP and SSR data demonstrated that the accessions had a similarity range from 0.65 to 1.00. Rough lemons and C. volkameriana accessions were relatively closely related. In lemon group, accessions from hybrid origin were distant from the others. We also applied principal components analysis (PCA) for a better presentation of relation among the accessions studies. Using PCA, 88.7% of the total variation in the original dimensions could be represented by just the two dimensions defined by the first two PCs. Although nearly all accessions could be distinguished, there was a low level of genetic diversity detected among lemon cultivars.
Citrus is one of the world's important fruit crops. Recently, citrus molecular genetics and biotechnology work have been accelerated in the world. Genetic transformation, a biotechnological tool, allows the release of improved cultivars with desirable characteristics in a shorter period of time and therefore may be useful in citrus breeding programs. Citrus transformation has now been achieved in a number of laboratories by various methods. Agrobacterium tumefaciens is used mainly in citrus transformation studies. Particle bombardment, electroporation, A. rhizogenes, and a new method called RNA interference are used in citrus transformation studies in addition to A. tumefaciens. In this review, we illustrate how different gene transformation methods can be employed in different citrus species.
ABSTRACT. 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 PCRbased DNA amplification procedure; 46 primers produced clear and good amplification. Ten of these 46 primers amplified monomorphic 2264 ©FUNPEC-RP www.funpecrp.com.br Genetics and Molecular Research 9 (4): 2263-2274 (2010) N. Comlekcioglu et al. 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.
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