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Öğe A PROMISING AND COST EFFECTIVE SURFACE STERILIZING METHOD FOR SWEET POTATO (IPOMOEA BATATASE.) CULTIVATED IN OPEN ENVIRONMENT(Parlar Scientific Publications (P S P), 2017) Bakhsh, Allah; Hussain, Tahira; Caliskan, Mehmet EminSweet potato is an important staple food and industrial material crop in the world. The crop is favored by resource poor farmers due to its good performance under adverse farming conditions, peak productivity in small farming areas and high carbohydrate and vitamin contents. The advances in genetic engineering technologies have helped the researchers to modify sweet potato to encode insect, herbicide and virus resistant traits. The efficient surface sterilization method is pre requisite for successful genetic improvement in sweet potato. We report an efficient, cost effective surface sterilization procedure of sweet potato seedlings collected from green house as well as field conditions for the initiation of in vitro culture. The seedlings from six sweet potato breeding lines were used to serve the purpose. Earlier, common bleach (NaOCl) was used to sterile sweet potato terminal buds to study their effectiveness in preventing microbial growth. Fungi and bacteria were most common microbial contaminants observed in cultures. Later on, the use of H2O2 in combination with ethanol resulted in relatively less contamination. The treatment of sweet potato ex plants with 70% ethanol for 5 min, 0.5% Mancozeb (Dithiocarbamate) for 5 min, followed by 5% hydrogen peroxide 5 mins resulted in suppressing microbial contaminations. This is the first report of the use of Mancozeb in combination with H2O2 for successful in vitro culture studies of sweet potato using explants from open environment. These findings will help to alleviate much burden associated with initiation of sweet potato cultures especially grown in field conditions.Öğe An insight into cotton genetic engineering (Gossypium hirsutum L.): current endeavors and prospects(SPRINGER HEIDELBERG, 2015) Bakhsh, Allah; Anayol, Emine; Ozcan, Sancar Fatih; Hussain, Tahira; Aasim, Muhammad; Khawar, Khalid Mahmood; Ozcan, SebahattinCotton (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.Öğe Development of potato lines resistant to colorado potato beetle ( CPB ) using RNAi-based transformation approach(Niğde Ömer Halisdemir Üniversitesi / Fen Bilimleri Enstitüsü, 2018) Hussain, Tahira; Çalışkan, Mehmet EminMost of the commercialized insect resistant transgenic crops express cry gene (s) isolated from Bacillus thuringiensis, however, intensive cultivation of Bt crops over almost two decades has been questioned regarding its sustainability and durability in pest management. The present study was focused to knock down highly specific molting associated Ecdysone receptor (EcR) gene of Colorado Potato Beetle (CPB) using RNAi approach. The partial cDNA of Ecdysone receptor gene of CPB was amplified using specific primers in sense and anti-sense orientation and cloned in pRNAi-GG vector flanked by an intronic sequence (pdk). Leaf and internodal explants of Agria and Lady Olympia cultivars were infected with Agrobacterium strain LBA4404 harboring plasmid pRNAi-CPB with EcR fragment under sense and antisense orientation under the control of 35S promoter. Neomycin phosphotransferase (nptII) gene was used as a plant selectable marker at a concentration of 100 mg L-1. The primary transformants obtained showed proper integration of T-DNA in plant genome by standard molecular analysis. The transgenic plants developed out of both cultivars were evaluated for their efficacy against first, second and third instar CPB larvae. The leaf biotoxicity assays revelead 15-80% mortality of CPB. A significantly lower fold change in larval weight was observed in insects fed on transgenic plants (0.87-4.14X) as compare to control (1.87-6.53X). Furthermore, CPB larvae fed on transgenic plants exhibited reduced transcripts of EcR gene indicating the functionality of EcR-dsRNA. The outcome of this research work is believed to reduce loses from insect pests in potato; thus leading to increased crop productivity.Öğe Development of Transgenic Potato Lines Expressing Ecdysone Receptor Gene of Colorado Potato Beetle(Springer, 2018) Bakhsh, Allah; Hussain, Tahira; Aasim, Muhammad; Pirlak, Ugur; Aksoy, Emre; Caliskan, Mehmet Emin[Abstract Not Available]Öğe Engineering crop plants against abiotic stress: Current achievements and prospects(UNITED ARAB EMIRATES UNIV, 2015) Bakhsh, Allah; Hussain, TahiraThe growth, development and productivity of crop plants is negatively influenced by abiotic stresses like drought, salinity, heat and chilling leading to significant losses in crop yield. The modern technology of genetic engineering has enabled the scientists to move genes from distant sources into crop plants to develop resistance against insect pests, weeds and invading pathogens, some of them have already been commercialized. Similarly, efforts have been made to develop crop plants with enhanced tolerance against drought, salinity and chilling and waterlogging stress. Engineering-crops against abiotic stresses has always been a challenge as this character is controlled by multigenes. The stress signaling and regulatory pathways have been elucidated using advanced molecular approaches and genes encoding tolerance to drought, salinity and chilling stress are being introduced in crop plants of economic importance using transformational technologies. The present review focuses the recent advances made in the development of transgenic crop plants of commercial importance with enhanced tolerance to abiotic stress; also the future prospects of stress tolerant crops have also been discussed.Öğe Mendelian segregation pattern and expression studies of insecticidal gene (cry1Ac) in insect resistant cotton progeny(UNITED ARAB EMIRATES UNIV, 2014) Hussain, Tahira; Bakhsh, Allah; Munir, Bushra; Hassan, Samina; Rao, Abdul Qayyum; Shahid, Ahmad Ali; Husnain, TayyabThe progenies of transgenic lines Bt-14 and Bt-17 developed as an independent transformation event from a local cotton variety CIM-482 harboring two insecticidal genes (cry1Ac & cry2a) were evaluated to determine resistance against lepidopterans, mainly Helicoverpa armigera L. under field conditions. The standard molecular techniques, i.e. polymerase chain reaction (PCR), enzyme-linked immunosorbent assay (ELISA) and western dot blot were used to confirm gene presence and expression level of transformed Bt gene, and its transfer pattern to further progeny. PCR confirmed integration of insecticidal gene in most of the plants in transgenic progeny, while expression of Bt gene quantified by ELISA and western dot blot showed variation in cry1Ac expression levels but interestingly, it conferred full protection against targeted insect pests. The leaf bioassays were conducted to determine the effectiveness of Bt genes against Helicoverpa armigera by calculating the mortality percentage of larvae. Most of the transgenic lines showed 70-100% mortality % age of Helicoverpa armigera. The agronomic characteristics of the transgenic lines were also recorded along with non transgenic control variety CIM-482. Morphological, agronomic and fibre data of these transgenic lines was recorded and analyzed statistically. Our results show that these transgenic lines (especially Bt-17 line) are promising cotton germplasm to be used in an efficient breeding programme.Öğe Transgenic potato lines expressing CP4-EPSP synthase exhibit resistance against glyphosate(Springer, 2020) Bakhsh, Allah; Hussain, Tahira; Rahamkulov, Ilhom; Demirel, Ufuk; Caliskan, Mehmet EminPotato crops are particularly vulnerable to weed competition from emergence to canopy closure and are subject to significant yield loss. Glyphosate is broad spectrum herbicide used to control weeds worldwide. In order to incorporate glyphosate resistant trait in four potato cultivars (Lady Olympia, Desiree, Agria and Granola), an efficient, cost effective, reproducible and stable Agrobacterium-mediated genetic transformation protocol was performed using leaf and internodal explants. Agrobacterium strain LBA4404 harboring newly modified recombinant binary vector pCAMHE-EPSPS containing EPSP synthase gene under the control of Cauliflower mosaic virus 35S promoter was used to infect explants. The overall transformation efficiency was 26.4%. Of the 280 plants transferred to greenhouse, 74 plants were found to be PCR positive with gene of interest. GUS histochemical, Southern blot, RT-qPCR, lateral flow dipstick assays confirmed integration and expression of EPSPS in primary transformants. The putative transgenic plants developed from these cultivars possessed enhanced resistance to glyphosate applications in T0 and first tuber generation. These transgenic potato lines could be used as source of germplasm for an efficient potato breeding program. Key message The article reports the development of potato transgenic lines with enhanced tolerance against glyphosate.
