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Agrobacterium-mediated genetic transformation approach allows for introducing novel genes in cotton (Gossypium hirsutum L.). Development of efficient regeneration and transformation protocol is very important for recalcitrant plants like cotton. In the present study, five-day-old germinated mature embryo parts especially embryonic axis, hypocotyl and plumule of cotton ‘Lashata’ were wounded and inoculated with A. tumefaciens strain GV2260 harbouring plasmid p35S-GUS-INT. The binary plasmid p35S-GUS-INT contains neomycin phosphotransferase II (NPTII) gene driven by nopaline synthase (NOS) promoter and β-glucuronidase (GUS) gene controlled by cauliflower mosaic virus (CaMV) 35S promoter. After inoculation, explants were co-cultivated in liquid and agar-solidified MS medium for 48 h in dark condition. Agar-solidified co-cultivation medium increased transformation frequency as compared to liquid medium. The putative primary transformants were confirmed with histochemical GUS assay, PCR and RT-PCR. However, comparing the three culture explant, the embryonic axis explants had significant difference with embryonic hypocotyl and plumule explants in terms of number and percentage (%) of GUS, PCR and RT-PCR positive plant. The total transformation efficiency was recorded as 3% with varying GUS expression levels.

Common bean (Phaseolus vulgaris L.), besides being an agricultural product that can be consumed as fresh vegetable, is a significant legume widely being planted in both Turkey and world. Because of having different usage areas, it is being considered as a valuable plant for human nutrition, trade and in many respects. In this study, we aimed at genetically characterization of the local and registered common bean genotypes and population structure of genotype groups belong to these common bean genotypes in Turkey. For this purpose, total 102 common bean genotypes including 93 local genotypes from 8 provinces, 7 cultivars and 2 reference cultivars were analyzed by 13 fluorescent SSR markers. As the result of the study, it was determined that the total SSR allele number was 192 and the average allele number was 14.8. While it was found that there were no synonymous genotypes, the highest heterozygosity rate was determined in three loci. Factorial correspondence analysis partially demonstrated substructure among common bean genotype groups. Structure analysis showed the same results as the Nm values and the Fst values. In the study, it was observed that SSR markers could be easily used in the molecular studies of common bean germplasm. The obtained results will be able to be used at the conservation, utilization of local common bean genetic resources and at the marker assisted selection studyings.

Pamuğun Türk ekonomisindeki yeri oldukça önemlidir. Pamuğun in vitro koşullardaki çoğaltım başarısı veya genetiğinin değiştirilmesi tohumların, bitkiciklerin ve farklı eksplantların ucuz, etkili ve tekrarlanabilir bir şekilde in vitro koşullara aktarılabilmesi için geliştirilecek protokollere bağlıdır. Bu çalışmayla farklı büyüme ortamlarının (değişen sukroz konsantrasyonu) ve çeşitli katılaştırıcıların (agar ve fitojel) yedi farklı pamuk çeşidinde in vitro çimlenme ve bitki gelişimi üzerindeki etkisine bakılmıştır. Sukroz içermeyen MS ortamındaki çimlenme oranının en yüksek olduğu gözlenmiştir. Ayrıca sukroz konsantrasyonundaki azalma ile çimlenme yüzdesindeki artış arasında doğru bir orantı vardır. Dahası, farklı konsantrasyonlarda katılaştırıcı içeren ortamlarla yapılan in vitro çimlenme deneylerinde en yüksek oran %0,4 fitojel içeren MS ortamında gerçekleşmiştir. Ayrıca bu oranın güçlü in vitro bitki gelişimini de sağladığı in vitro çoğaltım deneylerinde gözlenmiştir. Bu çalışmayla ilk defa sukroz içermeyen MS ortamının farklı pamuk çeşitlerindeki çimlenmeyi arttırdığı gösterilmiştir. Böylece, kısa sürede sağlıklı pamuk bitkileri elde edebilmek için en uygun ortamın % 0,4 (w/v) fitojel içeren sukrozsun MS ortamı olduğu söylenebilir., The significance of cotton can hardly be over emphasized in Turkish economy as it contributes momentously to agricultural value added products. The success of propagating cotton in in vitro conditions or to modify it genetically largely depends on an efficient, reproducible and cost effective protocol for in vitro germination of seeds, seedlings development and different explant sources. The present study was conducted to investigate the effect of different media (various sucrose concentrations) and gelling agents (Agar and Phytagel) on in vitro germination and seedling growth of different cotton cultivars. Results showed that MS medium devoid of sucrose contents induced maximum germination rates. The decreased sucrose concentration resulted in increased germination rates. Furthermore, experiments with different concentrations of gelling agents exhibited that MS media supplemented with 0.4 (w/v) % Phytagel results in induction of maximum in vitro germination rates; leading to robust seedling development suitable for further in vitro propagation experiments. According to our knowledge, this is the first report of increased germination rates of cotton cultivars incubated on MS media without the presence of sucrose; hence MS medium devoid of sucrose and solidified with 0.4% of Phytagel is most suitable for in vitro seed germination of cotton cultivars to obtained healthy seedlings in a shorter period.

Insect-resistant transgenic cotton has been commercialized for two decades. Most of the introduced cultivars express Bt gene(s) constitutively under the control of 35S promoter in whole-plant tissues. However, there have been other promoters considered by researchers to confine the toxin expression to targeted organ and tissues. We developed a triple-gene construct including GNA, cry1Ac and cp4 epsps genes. We attempted to confine cry1Ac expression to insect biting sites by cloning it to downstream of a wound-inducible promoter isolated from Asparagus officinalis (AoPR1). Moreover, to broaden the range of resistance, GNA was driven by the 35S promoter to target the sap-sucking insects like aphids which impose large losses in cotton production. To select the transformants in selection medium and for glyphosate tolerance, GNA and cry1Ac genes were accompanied with cp4 epsps gene. Two binary vectors harboring desired genes were constructed and utilized in the study (pGTGNAoC1AC and pGTGN35C1AC). Transformation of cultivar GSN-12 was carried out by employing Agrobacterium tumefaciens strain EHA105. Plantlets were primarily screened under glyphosate (N-phosphonomethyl glycine) selection pressure and subsequently subjected to molecular and biotoxicity assays. Introduction of cry1Ac and GNA to cotton plant conferred resistance to Spodoptera littoralis and Aphis gossypii Glover. Restriction of cry1Ac toxin protein to insect biting sites along with a plant lectin attributes significantly to insect pest management strategies. © 2018, Korean Society for Plant Biotechnology and Springer Japan KK, part of Springer Nature., Acknowledgements The PhD. fellowship awarded by The Scientific and Technological Research Council of Turkey (TUBITAK)-BIDEB to Dr. S.D. Khabbazi is deeply appreciated. The authors are grateful to the Leicester University (UK) for giving permission to use AoPR1 promoter for research purposes, Dr. Selma Onarıcı (TÜBİTAK GMBE) for providing pJIT61.cry1Ac plasmid and Prof. Umut Toprak (Department of Crop Protection, Ankara University) for providing S. littoralis larvae.

WOS:000352586600012 PubMed: 25735537 Opium poppy (Papaver somniferum) is an important medicinal plant producing benzylisoquinoline alkaloids (BIA). MicroRNAs (miRNAs) are endogenous small RNAs (sRNAs) of approximately 21 nucleotides. They are noncoding, but regulate gene expression in eukaryotes. Although many studies have been conducted on the identification and functions of plant miRNA, scarce researches on miRNA regulation of alkaloid biosynthesis have been reported. In this study, a total of 316 conserved and 11 novel miRNAs were identified in opium poppy using second-generation sequencing and direct cloning. Tissue-specific regulation of miRNA expression was comparatively analysed by miRNA microarray assays. A total of 232 miRNAs were found to be differentially expressed among four tissues. Likewise, 1469 target transcripts were detected using in silico and experimental approaches. The Kyoto Encyclopedia of Genes and Genomes pathway analyses indicated that miRNA putatively regulates carbohydrate metabolism and genetic-information processing. Additionally, miRNA target transcripts were mostly involved in response to stress against various factors and secondary-metabolite biosynthesis processes. Target transcript identification analyses revealed that some of the miRNAs might be involved in BIA biosynthesis, such as pso-miR13, pso-miR2161 and pso-miR408. Additionally, three putatively mature miRNA sequences were predicted to be targeting BIA-biosynthesis genes.

In plant tissue culture studies, obtaining new plantlets from different parts of plant is a very important feature with direct or indirect ways of regeneration. The plant show different regeneration capacities from species to species. In this present study, in-vitro direct shoots development from embryonic axis of cotton (Gossypium hirsutum L. cv. Lashata) were comparatively studied. Embryonic axis, embryonic hypocotyl and plumule obtained from five days old in-vitro grown seeds. All explants were cultured on MS media supplemented with 1- 4 mg/l Benzyl amino purine (BAP) + 0.1 mg/l Naphthalene acetic acid (NAA) and 3 mg/l activated charcoal. Cultured explants were kept at growth chamber with photoperiod for 16 hours light and 8 hours dark, at 25oC. Reproducible in-vitro plant regeneration was obtained from embryonic axis when cultured onto MS medium supplemented 1 mg/l BAP + 0.1 mg/l NAA and 3 g/l Activated charcoal. Shoot induction was to be noted 53.3%, 36.7% and 12.3 in embryonic axis, plumule and embryonic hypocotyl respectively. All these shoots were capable of rooting on MS medium supplemented with 0.1 mg/l NAA, and establishing in soil 3-4 weeks.

Grass pea (Lathyrus sativus L.) family leguminosae is cultivated as an important food and feed crop all over the world. It is very recalcitrant and difficult to regenerate and root under in vitro conditions. In this cotext, the study was carried out in three steps to find out the effects of three auxins [naphthalene acetic acid, indole 3 butyric acid, indole-3-acetic acid (IAA)], four sucrose concentrations and six types of substrate most suited for plant growth and helpful in acclimatisation of grass pea. The results showed that 2 mg L-1 IAA, 3 % sucrose was most suitable for rooting of grass pea. When different concentrations of sucrose were supplied to optimum concentration of IAA in Murashige and Skoog medium, 4.5 % sucrose concentration induced maximum number of 13.70 roots per explants that had positive impact on root length, fresh and dry weight of roots, plant height and morphology of the growing plants. There was 92.66 % acclimatisation and survival rate of these plants using peat moss compared to five other substrates used in this study. The developing plants were vigorous, flowered and set seed contained in pods under glass house conditions. It is concluded that rooting is affected by type and concentration of plant growth regulators and type of substrate has direct bearing on acclimatisation, flowering, pod and seed set of grass pea. As such this paper reports an efficient rooting and acclimatisation system of grass pea that will be very useful in future genetic transformation and breeding for improved characteristics.

The expression of insecticidal genes must be induced at appropriate time and in sufficient amount to confer protection against targeted pests. However, the increased scientific reports of resistance development in insect pest against insecticidal delta-endotoxins, produced by Bacillus thuringiensis, provide impetus for the development of alternative insect management strategies. The present study was conducted to investigate the importance of targeted expression of a hybrid insecticidal gene (SN19) in potatoes. For this purpose, two plant expression vectors were constructed by cloning hybrid SN19 gene (cry1Ba-domain I–III and cry1Ia-domain II) under the control of a wound-inducible promoter isolated from Asparagus officinalis (AoPR1) and CaMV 35S promoter, and were transferred to Agrobacterium tumefaciens strain EHA 105. Four potato genotypes (Marabel, Innovator, Tokat 10/1 and Tokat 6/24) were transformed with EHA 105 strain harboring pTF101.1 35S–SN19 and pTF101.1 AoPR1–SN19 constructs. Phosphinothricin (PPT) was used at concentration of 1 mg/l for selection of primary transformants. PCR results showed the presence of both introduced SN19 and bar genes in 43 plants out of total 154 putative transgenics. Expression of SN19 protein in primary transformants was confirmed by Western blot assays. The mechanical wounding of transgenic plants exhibited more accumulated levels of SN19 proteins during post wounding period. Leaf biotoxicity assays with Colorado potato beetle (Coleoptera) and tomato leafminer (Lepidoptera) exhibited 100% mortality of the pests in primary transformants. Based on our mortality results with both constructs, we concluded that the potato transgenic lines exhibited targeted expression of insecticidal gene under the control of AoPR1 promoter upon insect wounding with eliminated toxicity of Cry protein and hence can be further used effectively in potato breeding programme. © 2017, Korean Society for Plant Biotechnology and Springer Japan KK. 111T816 Türkiye Bilimsel ve Teknolojik Araştirma Kurumu, TÜBITAK Genetic Engineering and Biotechnology Institute, Scientific and Technological Research Council of Turkey, Gebze, Kocaeli, Turkey Funding The research project was supported by a Grant from Scientific and Technological Research Council of Turkey (TÜBİTAK) Project # 111T816.

WOS:000324281500012 Plant genetic engineering has opened new avenues to modify crops and has provided a powerful tool for crop improvement. The present study reports the development of regeneration and genetic transformation protocol for the Turkish cowpea cultivar Akkiz. The immature cotyledons were cultured on Murashige and Skoog (MS) medium containing 0.25-0.75 mg L-1 6-benzylaminopurine with or without 0.25 mg L-1 alpha-naphthalene acetic acid. Shoot regeneration varied 44.4%-83.3% with 2.1-5.0 shoots per explant. Regenerated shoots were rooted in MS medium containing 0.50 mg L-1 indole-3-butyric acid and acclimatized in the greenhouse, where they flowered and set seeds. Immature cotyledons were transformed with Agrobacterium tumefaciens strain LBA4404 harboring the recombinant binary vector pRGG containing an herbicide tolerance gene (bar) along with a uida (GUS) gene under 35S promoter. Phosphinothricin was used as a selectable marker at a concentration of 2.5 mg.L-1. Putative transformants were screened by-the histochemical GUS assay. Furthermore, molecular analysis revealed the presence of the introduced gene in the,genome of cowpea cultivar Akkiz. The selected transgenic plants showed a resistance to Basta (R) nonselective herbicide at up to 10 mL L-1 of water. Putative transgenic plants retained their pigmentation and continued to grow in the greenhouse.

Erysiphe trifolii and Leveillula taurica on Onobrychis spp. in Turkey, are described and illustrated.

Cowpea is an important grain legume crop. The study reports an efficient in vitro multiplication and shoots regeneration protocol from preconditioned embryonic axes of the Turkish cowpea cultivar Akkiz. The embryonic axes were preconditioned with 10 mg/l BA on agar solidified MS medium for 5 days. Thereafter they were cultured on MS medium containing 0.25, 0.50, 0.75 and 1.00 mg/l BA with or without 0.10 mg/l NAA. Mean frequency (%) of shoot regeneration, number of shoots per explant and shoot length decreased with each increase in BA concentration used singly. However, a positive increase was recorded in all parameters in the presence of 0.10 mg/l NAA in the regeneration medium. A maximum mean number of 10.33 shoots per explant was recorded on an MS medium containing 1.00 mg/l BA -0.1 mg/l NAA. Regenerated shoots were rooted on an MS medium containing 0.50 mg/l IBA. Rooted plants were acclimatized at room temperature in soil mix contained in pots where they were subjected to an intermittent mistwater spray for 24 h that maintained 90% relative humidity during the first few days which was gradually reduced to 40% for 10 days. All plants flowered and set seeds in a greenhouse after 3 months.

Expression of cry1Ac gene from Bacillus thuringiensis (Bt) was evaluated under the control of a wound-inducible AoPR1 promoter from Asparagus officinalis in transgenic tobacco plants. The leaves of transgenic plants were mechanically wounded to evaluate the activity of the AoPR1 promoter in driving the expression of Cry1Ac protein at the wound site. Our results indicate that mechanical wounding of transgenic plants was effective in inducing the expression of Cry1Ac protein. As a result of this induction, the accumulated levels of Cry1Ac protein increased during 6–72 h post-wounding period. The leaves of transgenic tobacco plants were evaluated for resistance against Heliothis virescens and Manduca sexta in insect bioassays in two different ways. The detached tobacco leaves were either fed directly to the insect larvae or they were first mechanically wounded followed by a 72 h post-wounding feeding period. Complete protection of mechanically wounded leaves of transgenic plants was observed within 24 h of the bioassay. The leaves of transgenic plants fed directly (without pre-wounding) to the larvae achieved the same level of protection between 24 and 72 h of the bioassay.

Amplified fragment length polymorphism (AFLP) markers were used to investigate genotypic variability among 34 alfalfa clones (M. sativa) using 15 primer combinations with restriction enzymes EcoRI, Pstl and Msel. 34 unique AFLP fragments were observed. The 15 primer pairs produced a total of 1002 fragments of which 460 were polymorphic. The number of polymorphic fragments detected per primer combination ranged 7 to 67. Furthermore, 22 clone-specific markers were also detected in the 13 clones. Data analysis was performed with NTSYSpc version 2.1 software. Genetic distance values ranged 5.9374 to 1.1453. Fifteen clones which showed the highest genetic variation were selected for producing synthetic variety of Alfalfa.

WOS:000370124000001 PubMed: 26909086 Alkaloids are diverse group of secondary metabolites generally found in plants. Opium poppy (Papaver somniferum L.), the only commercial source of morphinan alkaloids, has been used as a medicinal plant since ancient times. It produces benzylisoquinoline alkaloids (BIA) including the narcotic analgesic morphine, the muscle relaxant papaverine, and the anti-cancer agent noscapine. Though BIAs play crucial roles in many biological mechanisms their steps in biosynthesis and the responsible genes remain to be revealed. In this study, expressions of 3-hydroxy-N-methylcoclaurine 4'-methyltransferase (4'OMT) and reticuline 7-O-methyltransferase (7OMT) genes were subjected to manipulation to functionally characterize their roles in BIA biosynthesis. Measurements of alkaloid accumulation were performed in leaf, stem, and capsule tissues accordingly. Suppression of 4'OMT expression caused reduction in the total alkaloid content in stem tissue whereas total alkaloid content was significantly induced in the capsule. Silencing of the 7OMT gene also caused repression in total alkaloid content in the stem. On the other hand, over-expression of 4'OMT and 7OMT resulted in higher morphine accumulation in the stem but suppressed amount in the capsule. Moreover, differential expression in several BIA synthesis genes (CNMT. TYDC, 6OMT, SAT. COR, 4'OMT. and 7OMT) were observed upon manipulation of 4'OMT and 7OMT expression. Upon silencing and overexpression applications, tissue specific effects of these genes were identified. Manipulation of 4'OMT and 7OMT genes caused differentiated accumulation of BIAs including morphine and noscapine in capsule and stem tissues.

WOS: 000389669800002, Plant lectins have become efficient sources of insect resistance in crops. The present study was conducted to identify, amplify, clone and characterize the plant lectin gene GNA. The lectin, present in Galanthus nivalis (snowdrop), is an agglutinin toxic to hemiptera. The attempt was made to elucidate the relationship of the lectin gene trGNA (GNA isolated and characterized from Turkey) with other previously cloned lectins having insecticidal activity and to ensure the presence of the conserved mannose-binding region/site in the gene sequence. The full-length cDNA of trGNA was 477 bp that contained a 333 bp open reading frame encoding 157 amino acid proteins with 23 amino acids of signal peptide. BLAST results showed that trGNA has 89-97% similarity with previously reported GNA sequences while it has 84-96% similarity with earlier reported GNA protein sequences. No intron was detected within the region of genomic sequence corresponding to trGNA full-length cDNA. According to the search results from the NCBI (National Centrer for Biotechnology Information database), trGNA from Galanthus nivalis is most similar to the previously reported lectin sequences of Narcissus tazetta with a similarity percentage of 87%. The obtained results are useful for engineering of plants with enhanced insecticidal activity against chewing and sucking insects, causing crop pests. In addition, medical application of lectins may also be considered., Scientific and Technological Research Council of Turkey (TUBITAK) - BIDEB [2215], The authors are grateful to The Scientific and Technological Research Council of Turkey (TUBITAK) - BIDEB for granting PhD. fellowship (2215) to Mr. S.D. Khabbazi.

WOS: 000389771500008, In order to address biosafety concerns regarding the constitutive expression of foreign genes in crops, we applied a strategy aimed at confining foreign gene expression in insect wounding sites of cotton. For this purpose, a plant expression construct was designed by cloning the AoPR1 promoter (pathogenesis-related protein gene isolated from Asparagus officinalis) upstream from the insecticidal gene cry1Ac. The Turkish cotton cultivar cv. STN-468 was transformed using the Agrobacterium tumefaciens strain LBA4404 containing the recombinant binary vector pRD400 harboring cry1Ac under a wound-inducible promoter. The neomycin phosphotransferase (nptII) gene was used as a selectable marker at a concentration of 100 mg/L. The primary transformants were analyzed for T-DNA integration and expression using standard molecular approaches. The efficacy of insecticidal gene control of the AoPR1 promoter was investigated using leaf bioassays with 2nd instar larvae of Helicoverpa armigera and Spodoptera littoralis. Positive primary transformants from T-0 progeny were further raised under greenhouse conditions to obtain progeny (T-1). The introduced gene was properly inherited and expressed in T-1 progeny. The mechanical wounding of plants resulted in increased cry1Ac protein levels during 0-48 h of the wounding period. The transgenic lines exhibited appreciable levels of resistance against targeted insect pests in the leaf bioassays. The use of a wound-inducible promoter to drive insecticidal gene expression is a valuable insect resistant management strategy as gene expression will remain limited to the insect biting sites of plant and crop, food and environmental concerns can be minimized., TUBITAK (The Scientific and Technological Research Council of Turkey) [2216], The corresponding author is grateful to TUBITAK (The Scientific and Technological Research Council of Turkey) for providing postdoctoral fellowship Code: 2216) to carry out the research activities at University of Ankara. The authors acknowledge Dr. Umut Toprak, Associate Professor, Department of Plant Protection, Faculty of Agriculture, University of Ankara for providing Helicoverpa armigera for our lab bioassays experiment

Phylogenetic relationships were studied based on morphology and nuclear ribosomal DNA internal transcript spacer (nrDNA ITS) sequences in 49 populations of 36 Onobrychis taxa belonging to 5 sections (Dendobrychis, Lophobrychis, Onobrychis, Heliobrychis and Hymenobrychis) naturally grown in Turkey. The morphological data (46 quantitative characters) were standardized and submitted for PCA analysis before the performance of hierarchical clustering analysis. In addition, a 577-bp region of the nrDNA ITS sequences was compared to generate a dendrogram using the neighbor-joining method. According to the obtained results, the Onobrychis genus is clearly divided into the Onobrychis and Sisyrosema subgenera both molecularly and morphologically; however, in terms of molecular data, while the Onobrychis subgenus showed polyphyletic features, the Sisyrosema subgenus had monophyletic features. Additionally, long-wing taxa belonging to Dendobrychis and Onobrychis and O. crista-galli belonging to Lophobrychis located in the same clade in Onobrychis showed polyphyletic features. This finding was confirmed in terms of long-wing taxa as well as morphology and indicated that these three sections were entangled with each other under the Onobrychis subgenus.

WOS:000378689600017, t: The present study addressed the successful in vitro seed germination, bulblet regeneration, increased bulblet size, hardening, and acclimatization of Persian lily (Fritillaria persica). Seed germination rate was recorded as 86.7% and 96.7% after 2 and 3 months of cold treatment at 4 °C, respectively. Bulblet explants taken from a germination experiment were cultured on Murashige and Skoog (MS) medium supplemented with 0.5–2.0 mg/L thidiazuron (TDZ). Maximum bulblet regeneration frequency and bulblets per explants were achieved on a medium containing 2.0 mg/L TDZ. Leaf disc and leaf scale explants isolated from germinated seedlings were also cultured on MS medium containing 2–6 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) with a constant concentration of 0.2 mg/L kinetin. Both explants yielded the highest bulblet regeneration on MS medium containing 2 mg/L 2,4-D and 0.2 mg/L kinetin. A combination of 1.5 g/L agar and 1.6 g/L GELRITE resulted in a maximum bulblet diameter with a higher number of roots. The addition of 10 g/L NaCl to the culture medium resulted in the best root and leaf formation and secondary bulblet regeneration. Exposure of regenerated bulblets to 10 g/L NaCl at 4 °C for 2 months on a medium with 20 g/L sucrose was also found to be beneficial for acclimatization.

WOS: 000373886900005, Most of the commercialized Bt crops express cry genes under 35S promoter that induces strong gene expression in all plant parts. However, targeted foreign gene expression in plants is esteemed more important as public may be likely to accept 'less intrusive' expression of transgene. We developed plant expression constructs harboring cry1Ac gene under control of wound-inducible promoter (AoPR1) to confine Bt gene expression in insect wounding parts of the plants in comparison with cry1Ac gene under the control of 35S promoter. The constructs were used to transform four Turkish cotton cultivars (GSN-12, STN-468, Ozbek-100 and Ayhan-107) through Agrobacterium tumefaciens strains GV2260 containing binary vectors p35SAcBAR.101 and AoPR1AcBAR.101 harboring cry1Ac gene under control of 35S and AoPR1, respectively. Phosphinothricin (PPT) was used at concentration of 5 mg L-1 for selection of primary transformants. The primary transformants were analyzed for transgene presence and expression standard molecular techniques. The transformants exhibited appreciable mortality rates against larvae of Spodoptera exigua and S. littoralis. It was found that mechanical wounding of T (1) transgenic plants was effective in inducing expression of cry1Ac protein as accumulated levels of cry1Ac protein increased during post-wounding period. We conclude that use of wound-inducible promoter to drive insecticidal gene(s) can be regarded as a valuable insect-resistant management strategy since the promoter activity is limited to insect biting sites of plant. There is no Bt toxin accumulation in unwounded plant organs, seed and crop residues, cotton products and by-products, thus minimizing food and environmental concerns., Scientific and Technological Research Council of Turkey TUBITAK [111O254], The work on development of transgenic cotton in our laboratory is being supported by grants from Scientific and Technological Research Council of Turkey TUBITAK (Project No. 111O254). The authors acknowledge contribution and support of TUBITAK. The authors are also thankful to Leicester University (UK) for giving permission to use AoPR1 promoter for research purposes.

Randomly Amplified Polymorphic DNA markers (RAPD) were used to assess the hybrid identity of individuals sampled as Phlomis x termessi Davis. Out of 95 primers screened, 11 primers produced reproducible amplification patterns used for discrimination of P. x termessi and their parents. Eleven primers produced 81 bands. Forty two percent of the RAPD bands existed in parents. Of the 54 bands found in P. lycia, 19 were found only in this species and 7 of these were monomorphic. Similarly, of 57 RAPD bands observed in P. bourgaei, 18 were found only in P. bourgaei and 6 of these were monomorphic. Among hybrid individuals, 35 of the 73 markers were monomorphic. Fifteen of these existed in individual parents showing that parents were homozygous for these markers. Of the 35 monomorphic bands observed among hybrid individuals, 5 were present in the samples of one of the parents and completely absent from the samples of the other; therefore, additive inheritance is indicated. Of the 5 additive bands, 1 was inherited from P. bourgaei and 4 were inherited from P. lycia. Among 38 polymorhic markers observed in hybrid individuals, 9 were new and hybrid-specific. Pollen fertility was also investigated. Mean pollen fertility for P. lycia and P. bourgaei was 93% and 97% respectively. However, mean pollen fertility for hybrids was 65% (+/- 10.5).

This study reports the molecular characterization, polymorphism, and phylogenetic relationships of Triticum aestivum, T. dicoccoides, T. urartu, and T. monococcum ssp. boeoticum, obtained from different locations in Anatolia, using 33 primer combinations to generate amplified fragment length polymorphism (AFLP) patterns in 31 individual plant samples. The objectives of this work were to estimate the phylogenetic relationships between these species and to investigate the genetic distance as a result of ecological and climatic factors. The origin of the A genome of polyploid wheats is also discussed. Eight hundred and seventy-five AFLP fragments had polymorphic loci, 133 of which were unique to T. monococcum ssp. boeoticum, 66 were unique to T. urartu, and 141 were unique to T. dicoccoides. Analysis using the program POPGENE showed polymorphism levels of T. monococcum ssp. boeoticum, T. urartu, and T. dicoccoides of 42.63, 32.34, and 27.71%, respectively. No correlation between genetic distance and ecological or climatic factors was recorded in this study. Our results support the hypothesis that T. urartu is a diploid ancestor of T. dicoccoides and T. aestivum. © 2007 The Linnean Society of London, Botanical Journal of the Linnean Society, 2007, 153, 67–72.

Cotton (Gossypium hirsutum L.) is the most significant cash crop and backbone of global textile industry. The importance of cotton can hardly be over emphasized in the economy of cotton-growing countries as cotton and cotton products contribute significantly to the foreign exchange earnings. Cotton breeders have continuously sought to improve cotton’s quality through conventional breeding in the past centuries; however, due to limited availability of germplasm with resistant to particular insects, pests and diseases, further advancements in cotton breeding have been challenging. The progress in transformation systems in cotton paved the way for the genetic improvement by enabling the researchers to transfer specific genes among the species and to incorporate them in cotton genome. With the development of first genetically engineered cotton plant in 1987, several characteristics such as biotic (insects, viruses, bacteria and fungi) resistance, abiotic (drought, chilling, heat, salt), herbicide tolerance, manipulation of oil and fiber traits have been reported to date. Genetic engineering has emerged as a necessary tool in cotton breeding programs, strengthening classical strategies to improve yield and yield contributing factors. The current review highlights the advances and endeavors in cotton genetic engineering achieved by researchers worldwide utilizing modern biotechnological approaches. Future prospects of the transgenic cotton are also discussed.

Habitat destruction has resulted in the extinction of many plant species from the earth, and many more face extinction. Likely, the annual endemic Mediterranean knapweed (Centaurea tchihatcheffii) growing in the Golbasi district of Ankara, Turkey is facing extinction and needs urgent conservation. Plant tissue culture, a potentially useful technique for ex situ multiplication, was used for the restoration of this ill-fated plant through seed germination, micropropagation from stem nodes, and adventitious shoot regeneration from immature zygotic embryos. The seeds were highly dormant and very difficult to germinate. No results were obtained from the micropropagation of stem nodes. However, immature zygotic embryos showed the highest adventitious shoot regeneration on Murashige and Skoog (MS) medium, containing 1 mg l(-1) kinetin and 0.25 mg l(-1) NAA. Regenerated shoots were best rooted on MS medium containing 1 mg l(-1) IBA and transferred to the greenhouse for flowering and seed set. As such, the present work is the first record of in vitro propagation of critically endangered C. tchihatcheffii, using immature zygotic embryos, and is a step forward towards conservation of this indigenous species.

Sternbergia fischeriana is an endangered geophyte and therefore in vitro micropropagation of this plant will have great importance for germplasm conservation and commercial production. Bulb scale and immature embryo explants of S. fischeriana were cultured on different nutrient media supplemented with various concentrations of plant growth regulators. Immature embryos produced higher number of bulblets than bulb scales. Large numbers of bulblets were regenerated (over 80 bulblets/explants) from immature embryos on Murashige and Skoog (MS) medium supplemented with 4 mg l−1 6-benzylaminopurine (BA) and 0.25 mg l−1α-naphthaleneacetic (NAA) or 2 mg l−12,4-dichlorophenoxyacetic acid (2,4-D) after 14 months of culture initiation. Regenerated bulblets were kept at 5 °C for 5 weeks and then transplanted to a potting mixture.

Tobacco leaf disc explants were inoculated with Agrobacterum tumefaciens strain GV2260 carrying p35S GUS-INT to determine the influence of different co-cultivation temperatures (18 – 26 °C), periods (24 – 96 h) and media (solid and liquid) on transformation efficiency. Kanamycin-resistant shoots developed on leaf discs inoculated with Agrobacterium after 4 weeks of culture initiation. Regenerated shoots were excised and rooted in the basal medium supplemented with 100 mg dm −3 kanamycin and 250 mg dm −3 augmentin. The rooted plantlets were finally transferred to compost and confirmed by GUS assay and PCR analysis. The highest transformation frequency was achieved from the explants co-cultivated with A. tumefaciens in liquid medium for 48 h at 22 or 24 °C.

An efficient and reliable micropropagation system for Persian clover (Trifolium resupinatum L.) was developed using different explants and media. Node, hypocotyl and cotyledonary node explants were cultured on Murashige and Skoog (MS) medium supplemented with combinations of either 6-benzyladenine (BA) and indole-3-butyric acid (IBA) or BA, Kinetin (KIN) and IBA. Direct multiple shoots developed within 6 weeks in all explants in most media tested. The best shoot multiplication capacity was obtained from cotyledonary node explants on MS medium containing 7.1 μM BA and 1 μM IBA or 14.1 μM BA and 1 μM IBA. Elongated shoots were rooted on either MS medium alone or combination with different concentrations of indole-3-butyric acid (IBA), indole-3-acetic acid (IAA) and α-naphthaleneacetic acid (NAA). High rooting was achieved in half strength MS medium containing 8 μM IBA.

WOS:000356358200017 Hemianthus callitrichoides 'Cuba' is an ornamental aquatic plant used in water gardens and phytoremediation. A protocol for its rapid and reproducible shoot organogenesis from apical shoot explants was developed. Optimum culture conditions for shoot proliferation were tested in MS media containing different concentrations of 6-(gamma,gamma-dimethylallylamino)purine (2iP), 6-benzylaminopurine (BAP), 1-phenyl-3-1,2,3-thiadiazol-5-yl urea (TDZ), and gibberellic acid (GA(3)). Plant growth regulators significantly (P

The species of the section Oxytona Bernh., classified under the genus Papaver L., are of great importance due to their medical alkaloid contents such as morphine, codeine, and thebaine. A clearly established classification system is not available for Oxytona species. The aim of this study was to find relationships among 53 accessions belonging to 3 important Oxytona species, i.e. Papaver bracteatum Lindl., P. orientale L., and P. pseudo-orientale (Fedde) Medw., using 19 different morphological characters, 1 chemical character, and 15 random amplified polymorphic DNA (RAPD) markers. Nineteen characters, including bract and sepal leaves, petal marks, and capsule shape and size were analysed, and distinct morphological differences among accessions were identified. The thebaine content of the accessions was determined via HPLC analysis. Of the 53 accessions, 21 contained thebaine alkaloid ranging from trace amounts to 2.5% and the rest contained no thebaine. RAPD-PCR markers were used for analysis of accessions at molecular level. A total of 81 of bands generated by 15 successful RAPD-PCR reactions were detected as polymorphic. The polymorphism rate was approximately 85%. The combination of morphological, chemical, and molecular marker analyses resulted in a better classification for the section Oxytona., Papaver L. cinsi altında sınıflandırılan Oxytona Bernh. seksiyonu türleri morfin, kodein ve tebain gibi tıbbi alkaloit içeriklerinden dolayı büyük öneme sahiptirler. Oxytona seksiyonu türleri arasında belirgin bir sınıflandırma sistemi yoktur. Bu çalışmanın amacı Oxytona seksiyonundan Papaver bracteatum Lindl., P. orientale L. ve P. pseudo-orientale (Fedde) Medw. türlerine ait 53 materyali arasındaki ilişkiyi belirlemektir. Brakte ve sepal yaprakları, petal izleri, kapsül şekli ve buna benzer 19 karakter analiz edilmiş ve materyaller arasında belirgin morfolojik farklılıklar saptanmıştır. HPLC analizi yolu ile materyallerin tebain içerikleri belirlenmiştir. 53 materyalin 21’i % 0 (iz miktar) ile % 2,5 arasında tebain içerirken, geriye kalan 32 materyalin tebain içermediği saptanmıştır. Materyallerin moleküler düzeyde analiz edilmesi için RAPD-PCR markörleri kullanılmıştır. Başarılı 15 RAPD-PCR reaksiyonu ile üretilen 96 bandın 81’i polimorfiktir. Polimorfizm oranı yaklaşık % 85 olarak bulunmuştur. Morfolojik, kimyasal ve moleküler markörlerin birlikte kullanılması Oxytona seksiyonunda daha iyi bir sınıflandırma ile sonuçlanmıştır.

Efficient and reproducible shoot regeneration has been established from immature cotyledons and embryo axes of Hungarian vetch. Immature embryo axes showed higher regeneration capacity than immature cotyledons in most media tested. Immature cotyledons appeared to produce the highest frequency of shoot regeneration on 20 μM 6-benzylaminopurine (BA) and 2.5 μM α-naphthaleneacetic acid (NAA)-containing medium; whereas, shoot regeneration on immature embryo axes was best achieved on a medium supplemented with 5 μM each of BA and NAA. Regenerated shoots were excised and rooted in half-strength MS medium supplemented with 5 μM indole-3-butyric acid (IBA). Rooted plantlets were acclimatized to ambient conditions and later established under greenhouse conditions.

A procedure is described for the rapid and efficient adventitious shoot regeneration from leaflets, petioles and stems of field-grown sainfoin plants. All explants formed shoots on a range of media supplemented with 6-benzyladenine (BA) and α-naphthaleneacetic acid (NAA). Stem explants appeared to have better regeneration capacity than leaflet and petiole explants in most media tested. The highest frequency of shoot regeneration was achieved from stem segments on a medium containing 20 μM BA and 0.5 μM NAA. Regenerated shoots rooted in half-strength Murashige and Skoog medium containing 5 μM indole-3-butyric acid and later established well under greenhouse conditions.

An efficient in vitro bulblet production procedure from immature zygotic embryos of endemic and endangered Muscari muscarimi Medik. was described in the current study. Zygotic embryos were first isolated from immature seeds and cultured on different nutrient media compositions supplemented with various combinations of ?-naphthalene acetic acid (NAA), picloram, dicamba, 6-benzylaminopurine (BAP), and thidiazuron (TDZ). The best bulblet regeneration (59 bulblets per explant) was achieved in Murashige and Skoog (MS) medium containing 4 mg/L BAP and 0.5 mg/L NAA after 1 year of culture initiation. Regenerated bulblets were then transferred into MS medium without plant growth regulators for rooting. Bulblets produced well-developed root systems and increased their size on this medium after 2 months. All rooted bulblets were successfully transplanted into a potting mixture and acclimatized to ambient conditions. © TÜBİTAK.

WOS:000337274500007 Lathyrus sativus L. (grass pea) is an important grain legume crop grown all over the world. It is very recalcitrant and difficult to regenerate under in vitro conditions. The present study determined the effect of preconditioning of cytokinins [10 and 20 mg L-1 thidiazuron (TDZ) and 2-isopentenyl adenine (2iP)] and postconditioning of dissimilar levels of TDZ-indole-3-butyric acid (IBA) and 2iP-IBA on in vitro shoot multiplication using embryonic node explants. The results showed that preconditioning followed by postconditioning on Murashige and Skoog (MS) medium with 0.25, 0.50, 0.75, and 1.0 mg L-1 2iP or TDZ combined with 0.10 mg L-1 IBA had positive effects on shoot multiplication. It seemed as if the TDZ-preconditioned and TDZ-IBA-postconditioned explants were more regenerative compared to 2iP-preconditioned or 2iP-IBA-postconditioned explants. Maximum shoot induction (100%) was achieved on MS medium containing 1 mg L-1 TDZ and 0.10 mg L-1 IBA, with a mean number of 31.10 shoots per explant. In comparison to the other cytokinin tested, 2iP-preconditioned and 2iP-IBA-postconditioned explants had longer shoots, with a mean shoot length of 8.10 cm. Root initiation was observed on all cultures 4 weeks after the transfer of shoots with the best rooting on MS medium containing 2 mg L-1 IBA. The rooting response was 76.56% with a mean number of 8.03 roots per shoot and root length of 8.2 cm. The healthy plants were transferred to a greenhouse for acclimatization and exhibited 90% survival. It was concluded that TDZ preconditioning was necessary for grass pea multiplication along with TDZ-IBA postconditioning. This regeneration protocol may facilitate future genetic transformation and breeding studies to reduce beta-N-oxalyl-L-alpha,beta-di-amino propionic acid content in grass pea. Scientific and Technological Research Council of Turkey (TUBITAK)Turkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK) The researchers are grateful to the Scientific and Technological Research Council of Turkey (TUBITAK) for supporting this research work (2216-Research Fellowship Program for Foreign Citizens).

Seven genotypes of winter durum wheat (Triticum durum Desf.) were cultured to establish an efficient method of callus formation and plant regeneration from mature embryo culture, and to compare the responses of immature and mature embryo cultures. Immature embryos were aseptically dissected from seeds and placed, with the scutellum upwards, in dishes containing Murashige and Skoog's (MS) mineral salts and 2mg 2,4- dichlorophenoxyacetic acid (2,4-D) per litre. Calli and regenerated plants were maintained on 2,4-D-free medium. Mature embryos were moved slightly on the imbibed seeds. For callus formation, the seeds with moved embryos were placed, furrow downwards, in dishes containing 8 mg 2,4-D per litre. The developed calli and regenerated plants were maintained on the MS medium. Plants regenerated from both embryo cultures were vernalized and grown to maturity in soil. Variability was observed among the wheat genotypes tested for various culture responses in both explant cultures. Callus induction rate and regeneration capacity of callus were independent of each other. Mature embryos have a low frequency of callus induction but a high regeneration capacity. Considering availability, rapidity and reliability, this form of mature embryo culture can be used as an alternative method for immature embryo culture.

Immature cotyledons and embryo axes of sainfoin were cultured on Murashige and Skoog (MS) media supplemented with various concentrations of 6-benzylaminopurine (BAP) and a-naphthaleneacetic acid (NAA) to induce adventitious shoot regeneration. The highest frequency of shoot regeneration occurred following an initial callus growth on a MS medium containing 0.5 mg/l BAP and 2 mg/l NAA. Immature embryo axes showed higher regeneration capacity than immature cotyledons, however, shoot elongation was best achieved on immature cotyledons. Regenerated shoots were excised and rooted in half strength MS medium with 1 mg/l indole-butyric acid (IBA) or 1 mg/l NAA. The rooted plantlets were finally transferred to compost.

The hairy vetch (Vicia villosa Roth) is a climbing, prostrate or trailing legume grown as forage.It fixes atmospheric nitrogen, reduces soil erosion and provides an instant mulch. Multiple axillary shoot regeneration from a mature seed explant (zygotic embryo with two cotyledons) was obtained on MS medium containing 0.05 - 1.6 mg/l TDZ with or without 0.10 mg/l IBA. The frequency (%) of shoot regeneration ranged from 45.83-75.00% with a maximum number of 28.6 shoots per explant on MS medium containing 0.20 mg/l TDZ-0.10 mg/l IBA. The mean shoot length decreased proportionately with each increase in TDZ concentration irrespective of the IBA concentration in the culture medium. However, comparing the two types of regeneration media, longer shoots were recorded in the presence of IBA in the culture medium. Regenerated shoots were pulse treated with 50 mg/l IBA for 5, 10 and 20 min for rooting.

A rapid and simple method for adventitious shoot regeneration and somatic embryogenesis from immature cotyledon explants of pea (Pisum sativum L.) is described. Cotyledon size and the explant orientation to the medium surface were shown to have a clear effect on shoot regeneration. The highest frequency of shoot regeneration was achieved when the distal end of the greenest cotyledons (7–8 mm in size) were placed in contact with the agar surface. Shoots rooted at a frequency of 80–90% and grew into normal fertile plants. Somatic embryos were induced in cultures of immature cotyledons on modified MS medium containing high levels of α-naphthaleneacetic acid (27–215 μM) and 2,4-dichlorophenoxyacetic acid (23–181 μM). A higher frequency of somatic embryos with a normal morphology were induced using α-naphthaleneacetic acid.

There is much data to indicate that only a small number of cells in plant explants are competent for stable transformation by Agrobacterium. Circumstantial evidence suggests that certain cells reentering cell division at wound sites are competent for transformation by Agrobacterium. We have discovered a member of the intracellular PR gene family from asparagus (AoPR1) which is strongly expressed upon wounding and during the reactivation of the cell cycle in cultured asparagus cells, but which shows very little expression in intact plant tissues. The promoter from the AoPR1 gene was fused to an intron-containing GUS reporter gene and shown to be more strongly expressed than the commonly used CaMV 35S constitutive promoter in target cells for plant transformation. A transcriptional fusion of the AoPR1 promoter with an NPT-II gene was found to be a very efficient marker for the selection of transgenic tobacco callus. Expression of the AoPR1-NPT-II gene allowed efficient shoot formation on transgenic callus and efficient adventitious root formation on transgenic shoots. These latter observations provided firm evidence that transformation selection marker gene expression is most crucial at the early stages of the transformation process, during the establishment of transformed micro-calli.

Multiple shoot formation from the plumular apices excised from mature embryos of cowpea cv. Akkiz was obtained after pulse treatment with 10 mg/l BAP for 5 days followed by culture on MS medium containing 0.25, 0.50, 0.75 and 1.00, 1.25 mg/l BAP - with or without 0.10 mg/l NAA. Callus induction and shoot regeneration was recorded on all cultures containing BAP with or without NAA. However, inclusion of 0.1 mg/l NAA had positive effect on callus diameter and shoot length. Maximum mean number of 7.11 shoots per explant were obtained on MS medium containing 1.00 mg/l BAP. Longer shoots were recorded on MS medium containing various concentration of BAP + 0.1 mg/l NAA compared to those containing various concentrations of BAP singly. All shoots cultured on MS medium containing I mg/l BAP were rooted on MS medium containing 0.50 mg/l IBA. Rooted plants were acclimatized at room temprature in soil contained in pots. All plants flowered and set seeds in the greenhouse after 3 months. (C) 2009 Elsevier B.V. All rights reserved.

Seed protein diversity of fourteen lentil cultivars grown in Turkey was studied by using sodium dodecyl sulfate polyacrylamide gel electrophoresis. A distance matrix was produced based on five polymorphic protein bands, scored for their absence as 0 and presence as 1. Seed protein distances among the cultivars ranged from 0.00 to 0.80. The dendrogram based on the distance matrix indicated two distinct clusters. The first cluster includes the cultivars Sultan 1, Meyveci 2001 and Kayi 91. The second cluster contains the cultivars Pul 11, Ozbek, Emre 20, Malazgirt 89, Ciftci, Seyran 96, AliDayi, Firat 87, Sazak, Erzurum 89 and YerliKirmizi.

Randomly Amplified Polymorphic DNA markers were used to determine the genetic relationships among Turkish lentil cultivars and breeding lines. Fourteen cultivars and thirteen breeding lines were evaluated to determine genetic variability using nine random 10-mer primers (among 45 primers). Forty-one reproducible bands were obtained, 54% of which were polymorphic. Genetic distances among cultivars and breeding lines were obtained from the simple matching coefficients (SM). The lowest genetic distance was observed between the cultivars of Ali Dayi and Kafkas with 5.0%, while 'Seyran-96' and 'Ozbek' had the highest genetic distance with 58.3%. An unweighted pair-group method with arithmetic averages (UPGMA) cluster analysis was performed on the distance matrix using Phylip software. The dendrogram clearly showed two distinct groups. The first group is composed of Akm 565 and Akm 563. The second group contains all the cultivars and remaining eleven breeding lines.

Hypocotyl, cotyledon and cotyledonary node explants of Calendula officinalis L were cultured on Murashige and Skoog (MS) media supplemented with various concentrations of thidiazuron (TDZ), kinetin (KIN), α-naphthaleneacetic acid (NAA) and indole-3-butyric acid (IBA) to induce adventitious shoot regeneration and micropropagation. The highest frequency of adventitious shoot regeneration was achieved from hypocotyl and cotyledon explants on MS media supplemented with 0.75 mg dm−3 TDZ and either 0.25 or 0.50 mg dm−3 IBA. Efficient in vitro clonal propagation was also induced from cotyledonary nodes on a range of media supplemented with 0.75 mg dm−3 TDZ and 0.05 mg dm−3 NAA or 2 mg dm−3 KIN and 1 mg dm−3 NAA. Regenerated shoots were excised and rooted in MS medium supplemented with 1 mg dm−3 NAA. The rooted plantlets were finally transferred to pots.

Transformation of fenugreek ( Trigonella foenumgraecum) was carried out with A281 oncogenic strain of Agrobacterium tumefaciens using root, cotyledon and hypocotyl explants excised from 1-week-old seedlings, which showed that the plant was highly susceptible to transformation. Tumors ( calli) were selected on 50 mg dm(-3) kanamycin. They were analyzed for beta-glucuronidase (GUS) expression. Presence of uidA (gus) gene, was confirmed by polymerase chain reaction (PCR) amplification.

Efficient and reproducible shoot regeneration has been established from immature zygotic embryos of lesser burnet on MS medium containing 4 μM BAP and 10 μM NAA. Regenerated shoots were best rooted in half-strength MS medium supplemented with 10 μM NAA and later established in the greenhouse.

A procedure has been developed which allows high frequency adventitious shoot regeneration from immature cotyledons of pea. Prolific shoot regeneration occurred following an initial callus growth on a Murashige and Skoog (MS) medium containing 0.5 mg/l 6-benzylaminopurine (BAP) and 4 mg/l α-naphthaleneacetic acid (NAA). Cotyledon expiants proximal to the embryonic axis had the highest regeneration potential, however, the presence of an embryonic axis inhibited adventitious shoot regeneration. Addition of silver nitrate (AgNO3) to the medium did not promote the number of regenerated shoots but resulted in shoots with well developed tendrils and large stipules which had a reduced rooting capacity. Regenerated shoots rooted readily (80-90%) in half strength MS medium containing 1 mg/l indole-butyric acid (IBA) and further established well in compost.

Multiple shoots from shoot meristems of three - five-day-old in vitro grown seedlings of Turkish cowpea (Vigna unguiculata L.) cv. Akkiz was obtained in MS supplemented with 0.50 mg/l BAP - 0, 0.10, 0.30 and 0.50 mg/l NAA. Callus induction was recorded on all cultures. Callusing was recorded on all cultures containing 0.5 mg/l BAP with and without NAA. However, increased diameter of calli was recorded on MS medium containing 0.5 mg/l BAP - 0.1, 0.3 and 0.5 mg/l NAA. The highest frequency (%) of shoot regeneration and mean number of shoots per explant was recorded on MS medium containing 0.5 mg/l BAP with out NAA. Addition of any concentration of NAA resulted in significant decrease in the frequency (%) of shoot regeneration and mean number of shoots per explant. Maximum mean number of 2.60 shoots per explant was obtained on MS without NAA. Regenerated shoots were rooted on MS containing 0.50 mg/l IBA where up to seven adventitous secondary shoots arose from the base of mother shoot were also recorded. These shoots could also be rooted easily on the same rooting medium. Rooted plants were adapted at room temprature in soil mix in pots. All plants flowered and set seeds in the growth room after three months. Key words: Cowpea, Micropropagation, Shoot meristem doi:10.3329/bjb.v37i2.1723 Bangladesh J. Bot. 37(2): 149-154, 2008 (December)