Allah BAKHSH, Muhammet Çağrı OĞUZ, Surendra BARPETTE, Ömer Can KARAKOÇ, Sebahatin ÖZCAN, Emine ANAYOL, Sancar Fatih ÖZCAN

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

The 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.

PDF

___

Ahmed HA, Hajyzadeh M, Barpete S, Özcan S (2014). In vitro plant regeneration of Iraqi cotton (Gossypium hirsutum L.) cultivars through embryonic axis. J Biotechnol Res Cent (special edition) 8: 90-94.Bajwa KS, Shahid AA, Rao AQ, Kiani MS, Ashraf MA, Dahab AA, Bakhsh A, Latif A, Khan MAU, Puspito AN et al. (2013). Expression of Calotropis procera expansin gene CpEXPA3 enhances cotton fiber strength. Aus J Crop Sci 7: 206-212.Bakhsh A, Anayol E, Özcan S (2014). Comparison of transformation efficiency of five Agrobacterium tumefaciens strains in Nicotianatabacum L. Emir J Food Agr 26: 259-264.Bakhsh A, Emine A, Özcan SF, Hussain T, Aasim M, Khawar KM, Özcan S (2015). An insight into cotton genetic engineering (Gossypium hirsutum L.): current endeavors and prospects. Acta Physiol Plant 37: 171-179. Bakhsh A, Rao AQ, Shahid AA, Husnain T, Riazuddin S (2010). Camv 35S is a developmental promoter being temporal and spatial in expression pattern of insecticidal genes (Cry1ac & Cry2a) in cotton. Aust J Basic Appl Sci 4: 37-44.Bakhsh A, Shahzad K, Husnain T (2011). The variation in spatio temporal expression of insecticidal genes in transgenic cotton. Czech J Genet Plant Breed 47: 1-9.Bakhsh A, Siddiq S, Husnain T (2012). A molecular approach to combat spatio-temporal variation in insecticidal gene (Cry1Ac) expression in cotton. Euphytica 183: 65-74.Barampuram S, Allen G, Krasnyanski S (2014). Effect of various sterilization procedures on the in vitro germination of cotton seeds. Plant Cell Tiss Organ Cult 118: 179-185.Barpete S, Aasim M, Khawar KM, Özcan SF, Özcan S (2014a). Preconditioning effect of cytokinins on in vitro multiplication of embryonic nod of grass pea (Lathyrus sativus L.). Turk J Biol 38: 485-492. Barpete S, Anayol E, Ahmed HA, Özcan SF, Köm D, Delpasand S, Aasim M, Khawar KM, Özcan S (2013). Optimization of in-vitro culture condition for genetic transformation of cotton (Gossypium hirsutum L). In: The International Plant Breeding Congress (IPBC), Antalya, Turkey, pp. 248. Barpete S, Khawar KM, Özcan S (2014b). Differential competence for in vitro adventitious rooting of grass pea (Lathyrus sativusL.). Plant Cell Tiss Organ Cult 119: 39-50.Barpete S, Oğuz MC, Özcan SF, Anayol E, Ahmed HA, Khawar KM, Özcan S (2015a). Effect of temperature on germination, seed vigor index and seedling growth of five Turkish cotton (Gossypium hirsutum L.) cultivars. Fresenius Environ Bull 24: 1-7.Barpete S, Özcan SF, Aasim M, Özcan S (2015b). In vitro high frequency regeneration through apical shoot proliferation of Hemianthus callitrichoides Cubaa multipurpose ornamental aquatic plant. Turk J Biol 39: 493-500. Breitler JC, Vassal JM, Catala MDM, Meynard D, Marfa V, Mele E, Royer M, Murillo I, San Sequndo B, Guiderdoni E et al. (2004). Bt rice harbouring cry genes controlled by a constitutive or wound-inducible promoter: protection and transgene expression under Mediterranean field conditions. Plant Biotechnol J 2: 417-430.Chen S, Wu J, Zhou B, Huang J, Zhang R (2000). On the temporal and spatial expression of Bt toxin protein in Bt transgenic cotton. Acta Gossypii Sin 12: 189-193. DOnofrio C, Morini S (2002). Increasing NaCl and CaCl2concentrations in the growth medium of quince leaves: II: effects on shoot regeneration. In Vitro Cell Dev Biol - Plant 38: 373-377.Daud M, Ali S, Variath MT, Zhu SJ (2012). Antioxidative enzymes status in upland cotton callus culture under osmotic stresses. In: The International Conference on Computational Techniques and Artificial Intelligence (ICCTAI-2012), Penang, Malaysia, pp. 279-282.Doyle J (1991). DNA protocols for plants-CTAB total DNA isolation. In: Hewitt GM, Johnston A, editors. Molecular Techniques in Taxonomy. Berlin, Germany: Springer-Verlag, pp. 283-293.Gould J, Magallanes-Cedeno M (1998). Adaptation of cotton shoot apex culture to agrobacterium-mediated transformation. Plant Mol Biol Rep 16: 1-10.Gulbitti-Onarici S, Zaidi MA, Taga I, Ozcan S, Altosaar I (2009). Expression of Cry1Ac in transgenic tobacco plants under the control of a wound-inducible promoter (AoPR1) isolated from Asparagus officinalis to control Heliothis virescens and Manduca sexta. Mol Biotechnol 42: 341-349.Han GY, Wang XF, Zhang GY, Ma ZY (2009). Somatic embryogenesis and plant regeneration of recalcitrant cottons (Gossypium hirsutum). Afr J Biotechnol 8: 432-437.Huang WL, Lee CH, Chen YR (2012). Levels of endogenous abscisic acid and indole-3-acetic acid influence shoot organogenesis in callus cultures of rice subjected to osmotic stress. Plant Cell Tiss Organ Cult 108: 257-263.Huang WL, Liu LF (2002). Carbohydrate metabolism in rice during callus induction and shoot regeneration induced by osmotic stress. Bot Bull Acad Sin 43: 107-113.Huang WL, Wang YC, Lee PD, Liu LF (2006). The regenerability of rice callus is closely related to starch metabolism. Taiwan J Agric Chem Food Sci 44: 100-107. James C (2014). Global status of commercialized biotech/GM crops: ISAAA Brief No. 49 ISAAA, Ithaca, NY, USA.Jimenez VM (2005). Involvement of plant hormone and plant growth regulators on in vitro somatic embryogenesis. Plant Growth Regul 47: 91-110.Khan GA, Bakhsh A, Ghazanfar M, Riazuddin S, Husnain T (2013). Development of transgenic cotton pure lines harboring a pesticidal gene (cry1Ab). Emir J Food Agr 25: 434-442.
Khan GA Bakhsh A, Riazuddin S, Husnain T (2011). Introduction of cry1Ab gene into cotton (Gossypium hirsutum) enhances resistance against Lepidopteran pest (Helicoverpa armigera). Span J Agr Res 9: 296-300. Khan ZA, Abdin MZ, Khan JA (2015). Functional characterization of a strong bi-directional constitutive plant promoter isolated from Cotton leaf curl Burewala virus. PLoS ONE 10(3): e0121656.Kumar M, Shukla AK, Singh H, Tuli R (2009). Development of insect resistant transgenic cotton lines expressing cry1EC gene from an insect bite and wound inducible promoter. J Biotechnol 140: 143-148.Lee ST, Huang WL (2014). Osmotic stress stimulates shoot organogenesis in callus of rice (Oryza sativa L.) via auxin signaling and carbohydrate metabolism regulation. Plant Growth Regul 73: 193-204. Liza LN, Nasar ANM, Zinnah KMA, Chowdhury MA, Ashrafuzzaman M (2013). In vitro growth media effect for regeneration of tomato (Lycopersicon esculentum) and evaluation of the salt tolerance activity of callus. J Agr Sustain 3: 132-143.Mahon R, Finnergan J, Olsen K, Lawrence L (2002). Environmental stress and the efficacy of Bt cotton. Aust Cotton Grower 22: 18-21.Marschner H (1995). Mineral Nutrition of Higher Plants. 2nd edition. San Diego, CA, USA: Academic Press, pp. 889.Mengle K (1980). Effects of potassium on assimilate conduction to storage tissue. Ber Beutsch Bot GCs 93: 353-362. Mishra R, Wang HY, Yadav N, Wilkins T (2003). Development of a highly regenerable elite Acala cotton (Gossypium hirsutum c v. Maxxa)a step towards genotype-independent regeneration. Plant Cell Tiss Organ Cult 73: 21-35.Molassiotis AN, Sotiropoulos T, Tanou G, Kofidis G, Diamantidis G, Therios I (2006). Antioxidant and anatomical responses in shoot culture of the apple rootstock MM 106 treated with NaCl, KCl, mannitol or sorbitol. Biol Plantarum 50: 61-68.Murashige T, Skoog F (1962). A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15: 473-97. Ochatt SJ, Conreux C, Jacas L (2013). Flow cytometry distinction between species and between landraces within Lathyrus species and assessment of true-to-typeness of in vitro regenerants. Plant Sys Evol 299: 75-85.Rao AQ, Bakhsh A, Nasir IA, Riazuddin S, Husnain T (2011). Phytochrome B mRNA expression enhances biomass yield and physiology of cotton plants. Afr J Biotechnol 10: 1818-1826.Satyavathi VV, Prasad V, Gita LB, Sita LG (2002). High efficiency transformation protocol for three Indian cotton varieties via Agrobacterium tumefaciens. Plant Sci 162: 215-223.Snedecor SW, Cochran WG (1967). Statistical Methods. Ames, IA, USA: The Iowa State University Press, pp. 327-329.Thomas TD (2008). The role of activated charcoal in plant tissue culture. Biotechnol Adv 26: 618-631.Tohidfar M, Ghareyazie B, Mosavi M, Yazdani S, Golabchian R (2008). Agrobacterium-mediated transformation of cotton (Gossypium hirsutum) using a synthetic cry1Ab gene for enhanced resistance against Heliothis armigera. Iranian J Biotechnol 6: 164-173.Watanabe HY (1970). Effects of nitrogen phosphorus and potassium on rice in relation to ascorbic acid and NAA on phosphorylation in rice in relation to photosynthetic rate of single leaves. Soil Sci Plant Nutr 16: 163-166.