In situ localisation of axi 1 (Auxin independent) Gene in Rhizobium leguminosarum inoculated root tips of pea plants [Axi 1 (Oksinden Baimsiz) Geninin, Rhizobium leguminosarumbv., pre ile Aşanmiş Bezelye Bitkisinin Kök Uçlarindaki in situ lokalizasyonu]

Axi 1 is the only characterized auxin independent gene in tobacco plants. In this study, the effect of axi 1 gene on cortical cell division during different stages of nitrogen fixation and on the role of polar auxin transport were investigated. Root and nodule samples were taken from pea seedlings 24 hours, 48 hours, 6 days and 10 days after inoculation with R. leguminosarum PRE strain. Using in situ hybridisation, the location of the axi 1 gene expression in root and nodule tissues was identified. The results showed that in root tissues inoculated with Rhizobia, the axi 1 gene is expressed on the phloem tissue of the vascular bundle. In the 10-day-old nodule tissues, the gene expression was found in the phloem tissue, around the nodule bundle and nodule meristematic region. The results obtained indicate that also Axi 1 gene plays a role in nitrogen fixation and auxine transport

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

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

Morphologic and histologic analysis of regeneration in some melon varieties

In this study, morphological and anatomic structures of in vitro regeneration for cotyledon explants belong to different melon genotypes were investigated. Regeneration was obtained from cotyledon explants using NA8 medium for Kirkagac 637 and Ananas which belong to Inodorus and Reticulatus group, respectively. Morphologically, it was shown that the formation of first swelling (7day), meristematic structure (12day) and shoot (17day) in Kirkagac (Inodorus group) was earlier than Ananas variety (Reticulatus group). In Ananas, first swelling, meristematic structure and shoot formation were obtained from 9, 15 and 20 day old tissues, respectively. Histologically, first meristematic structure was observed from 10 day old tissue in Kirkagac and 14 day old tissue in Ananas on a base of paranchymal cells among protuberances. © 2003 Taylor and Francis Group, LLC

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

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

Clonal propagation of disease-free rootstocks for sour and sweet cherry by meristem culture

Four different rootstocks were cultured, namely Primus mahaleb L, Gi-Sel Al (P.fruticosa Pall.X P. avium L.), Damil (P. dawyckensis Sealy) and Tabel (Edabriz). Plants were inspected for symptoms from mid-April through mid-May. Leaf samples from Prunus mahaleb and Gi-Sel Al were collected in the middle of May for ELISA and sap transmission tests. Damil and Tabel were not tested because they had already been certified. Meristems from both axillary and terminal buds were cultured in spring (May), summer (July-August) and autumn (September- October). ELISA and sap transmission tests did not give specific reactions for the viruses. P. mahaleb and Gi-Sel Al were classed as healthy rootstocks. Spring was the best time for culturing meristems. 92 % of meristems of P. mahaleb survived, while Gi-Sel Al explants did not proliferate and showed vitrification and callus induction after the second and third subcultures. Explants of Damil did not even proliferate and most were dead 2-3 weeks after the establishment of culture. In summer, survival of meristems of all plants was considerably less whereas bacterial contamination was very high, ranging from 17 %- 84 %. Meristems of all plants showed the lowest survival in autumn. In general, plants were subcultured every 3 or 4 weeks. During the rooting phase, only P. mahaleb plants were successfully rooted in culture media having 0.5-1.0 mg/1 IBA. 50 % of the explants showed good rooting capacity in 0.5 mg/1 IBA, 75-85 % of them with 1.0 mg/1 IBA. Roots were vigorous in both cases

Genetic transformation in citrus

PubMedID: 23983635Citrus 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. © 2013 Dicle Donmez et al

The genetic characterization of Turkish watermelon (Citrullus lanatus) accessions using RAPD markers

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. © 2010 Springer Science+Business Media B.V.TOVAG 104O073Acknowledgments The authors are grateful to the Council of Scientific and Technological Research of Turkey for the financial support (Project No: TOVAG 104O073). In addition, authors thank Dr. Sedat Serce for his kindly help for the statistical analyses of this research

Transformation of Cm-ADH gene to melon genotype

Alcohol dehydrogenases (ADH) participate the biosynthetic pathway of aroma volatiles in fruit by interconverting aldehydes to alcohols and providing substrates for the formation of esters. Two highly divergent ADH genes of cantaloupe Chareiitais melon (Cucumis melo var. cantalupensis 'Vedrantais') have been isolated. This research is aimed at identifying the role of the Cm-ADH genes (Cm-ADH1 and 2) in the synthesis of aroma volatiles via the RNAi method. Different regeneration media were investigated to improve transformation efficiency. The best regeneration result was obtained from MS medium supplemented with 2.2 M BAP and 0.54 NAA. Regeneration protocol has been applied to obtain transformed plants, using Agrobacterium tutriefaderis which has a vector RNAi pFGC5941-(11406 bp) carrying genes of resistance to kanamycifie and Bar. Transformed plants were characterized by PCR analysis