In vitro shoot regeneration from preconditioned explants of chickpea (Cicer arietinum L.)

The present study reports the successful shoot regeneration of preconditioned mature embryo and embryonic axis explants of chickpea cv. Gokce. Explants were preconditioned with 10 mgl benzylaminopurine (BA) for 7 days followed by culture on Murashige and Skoog (MS) medium containing 0.25, 0.50, 1.00 and 2.00 mg/l BA with or without 0.25 mg/l naphthalene acetic acid (NAA) supplemented with 4 mg/l activated charcoal and 1 mg/l polyvinylpyrrolidon (PVP). Shoot regeneration was recorded on all explants. Maximum number of (14.75) shoots per explants on mature embryo were recorded on MS medium containing 2.0 mg/l BA with 0.25 mg/l NAA. Whereas, 16.83 shoots per explants were recorded on MS medium containing 2.0 mg/l BA. Presence of NAA in the culture medium decreased the mean shoot length of both explants compared to medium devoid of 0.25 mg/l NAA. Regenerated shoots were rooted on MS medium containing 1.0 mg/l indole-butyric acid (IBA) after 4 weeks of culture. Rooted plantlets were transferred to pots for acclimatization under green house conditions

Molecular Characterization of Snowdrop Lectin (GNA) and its Comparison with Reported Lectin Sequences of Amaryllidaceae

WOS: 000389669800002Plant 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

Inducing osmotic stress leads to better genetic transformation efficiency in cotton (Gossypium hirsutum L.)

WOS: 000378689600012The present study investigated the effect of different salts on cotton shoot regeneration and transformation efficiency. Two-day-old germinating embryos of a local cotton cultivar (SG-125) were pretreated with 50 mM each of NaCl, CaCl2, and KCl for 60 min. The embryo explants were transformed by cocultivation with Agrobacterium tumefaciens strain LBA 4404 harboring a binary plasmid pTF101.1 that carried the insecticidal gene (cry1Ac) under control of wound-inducible promoter (AoPR1) and bilanafos acetyl reductase (bar) gene for plant selection. The salt-pretreated embryos showed maximum response on regeneration MS medium containing 0.50 mg/L 6-benzylaminopurine (BAP) and 0.10 mg/L indole-3-butyric acid (IBA), also supplemented with 5 mg/L bialaphos for in vitro screening of the transformed plantlets. The primary transformants were further screened by molecular techniques for integration and expression of the introduced gene. Maximum transformation efficiency (1.10%) was noted on KCl-treated explants compared to nontreated (control) explants. In conclusion, pretreatment of explants with 50 mM KCl for 60 min induced positive effects and triggered shoot regeneration in primary cotton transformants.Scientific and Technological Research Council of Turkey (TUBITAK) [111O254]The authors are grateful to the Scientific and Technological Research Council of Turkey (TUBITAK) for financial support of the research work (TUBITAK project no.: 111O254). The authors are also grateful to Prof Khalid Mahmood Khawar for his valuable suggestions for the improvement of the manuscript

EFFICIENT REGENERATION AND AGROBACTERIUM TUMEFACIENS MEDIATED GENETIC TRANSFORMATION OF POTATO (SOLANUM TUBEROSUM L.)

WOS: 000436522600048Potato (Solanum tuberosum L.) is highly demandable world food crop and has become a staple food in many parts of the world including Turkey, India and China. Potato under in vitro culture condition is recalcitrant and genotype dependent. Therefore, we tested several factors to enhance regeneration and genetic transformation efficiency of Turkish potato genotypes. The influence of physical factors and conditions in Agrobacterium mediated transformation of the potato leaves was tested. Using the best combination (10 mg L-1 ZR, 10 mg L-1 NAA) with Agrobacterium-mediated transformation, we regenerated independent putative transformants 61.6% of the total explants cultured. GUS and nptII positive plants were confirmed by molecular analysis (PCR) that showed transformation rate of 24.16%. The protocol described here is simple, efficient, and produces transgenic plant in 7-8 weeks after inoculation with Agrobacterium bacterium.Scientific and Technological Research Council of Turkey (TUBITAK)Turkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK)The present work is partially supported by the Scientific and Technological Research Council of Turkey (TUBITAK). The authors would like to thank Prof. Dr. Gungor Yilmaz (University of Gaziosmanpasa, Department of Field Crops) and Dr. Huseyin Onaran (Nigde Potato Research Institute) for providing the plant material. The authors would like to thank Dr Jitendra Kumar (Department of Plant Pathology, University of Minnesota, USA) for valuable suggestion and improvement of the present manuscript

CALLUS INDUCTION AND REGENERATION OF ALKANNA ORIENTALIS VAR. ORIENTALIS AND A. SIEHEANA

WOS: 000486533500028Callus induction and proliferation of Alkanna orientalis var. orientalis and Alkanna sieheana containing valuable alkannin/shikonin (A/S) derivates were investigated using leaf base and stem segment explants. Stem segments and cotyledonary leaf base of both species were cultured on Murashige and Skoog medium fortified with different concentrations of BAP, Kn, NAA, IAA and IBA for callus induction and shoot regeneration. High frequency reproducible, prolific and compact calli formation was obtained from the stem segments of both species in all media tested. The frequency variations of callus induction and shoot regeneration were discussed in terms of different species, plant growth regulators and explant resources. A. orientalis and A. sieheana may be considered to be alternative plants for the A/S production in vitro

DEVELOPMENT OF INSECT-RESISTANT COTTON LINES WITH TARGETED EXPRESSION OF INSECTICIDAL GENE

WOS: 000389771500008In 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 experimen

Towards better insect management strategy: restriction of insecticidal gene expression to biting sites in transgenic cotton

WOS: 000373886900005Most 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

Inter and intra annual variation in body condition of the Black Sea anchovy, Engraulis encrasicolus ponticus-Potential causes and consequences

Variations in the relative condition (Kn) of the Black Sea anchovy (Engraulis encrasicolus ponticus) have been evaluated over eleven overwintering seasons (2005-2016). During the study period, a linear decrease was observed in the condition each year. Difference in condition from the onset to end of winter varied between 10% and 20%. Overall however, condition displayed an increasing trend within the time frame studied. Considering the worsening state of the stock during the same period, this increasing trend indicates that condition may be dependent on the fish density. The negative relationship observed between body condition of anchovies and the spawning stock biomass supports the same view and points out that particularly in winter when food supply is limited, interspecies competition could be a considerable factor determining condition. The strong correlation between condition and eddy kinetic energy underlines that velocity of water movement might be an effective factor conserving energy use and consequently supporting maintenance of body condition. Finally, the positive correlation observed between condition loss during winter and number of recruits the following year indicates that how fish overwinter plays an important role in recruitment success rates