Assessment the Response of Chickpea Genotypes to Agrobacterium -Mediated Transformation System
Transformation protocol based on the inoculation of chickpea mature embryos with Agrobacterium suspension was carried out. Four chickpea lines and one Iraqi local variety were used as recipient to the foreign gene of Agrobacterium tumefaciens strain (AgL1). Three plasmids were already inserted in the bacteria cells. The first plasmid carries the bar gene coding for phosphinothricin acetyle transferase (PAT), which confers resistance to the herbicide phosphinothricin or glofosinate ammonium(PPT) and uidA (gusA) gene coding for β-glucuronidase (GUS). The other two plasmids carried the LeEREBP gene which confers drought resistance and bar gene coding for phosphinothricin acetyle transferase (PAT). Successfully regenerated explants were subjected to selection pressure on 10 mg /l of phosphinothricin PPT and the putative transgenic explants were rooted on root induction medium consisting of MS basal medium with B5 medium vitamins supplemented with 2.5 ml of 1mg /ml IBA in addition to grafting on 7 days old non- transformed rootstock. PCR approved transgenic chickpea. 600 mg/l of PPT was used by painting the leaves of surviving plants to detect the expression of bar gene which encodes for phosphinothricin acetyle transferase and confirmed herbicide resistance in transgenic plants.
Driver J.A. and Kuniyuki A.H.(1984). In vitro propagation of paradox walnut rootstock. Hort. Science 19: 4.
Fontana, GS. Santini , L; Caretto, S. ; Frugis, G. and Mariotti, D. (1993) Genetic transformation in the grain legume Cicer arietinum (chickpea). Plant Cell Rep. 12:194-199.
Gelvins, S.B,(2003) Improving plant genetic engineering by manipulating the host ,Trend in biotechnology ,21,95-98.
ICARDA, 2003. ICARDA Annual Report (2002). International Center of Agricultural Research in the Dry Area. Aleppo . Syria . IV:136pp.
Jefferson, R. A. (1987). Assaying chimeric genes in plants : the GUS gene fusion system.Plant molecular Biology Reporter. 8: 387-450.
Kar S. ; Basu, D.; Ramakrishnan N.A.; Mukhrejee , P.; Johnson, T. M.; Nayak P. and Sen S.K. (1997) expression of cry 1A(c) gene of Bacillus thuringiensis in transgenic Chickpea plants inhibits development of pod borer (Heliothis armigera) larvae, transgenic Research. 6: 177-185.
Kiesecker H. (2000). Entwicklung eines Agrobacterium tumefacienes vermittenlten gentransfersystems für kichererben (Cicer arietinum L. ). Ph.D. thesis , Hanover University, Germany, 114pp.
Krishnamurthy K.V.; Suhasini K.; Sagare A. P. ; Meixner M.; Dekathen A.; Picardt T. and Shieder O.(2000). Agrobacterium mediated transformation (Cicer arietinum L.) embryo axis. Plant Cell Report . 19: 235-240.
Murashige, T. and Skoog, F (1962)A revised method for rapid growth and bioassays with tobacco tissue culture. Physiol Plant 15:473-497.
Pooran M Gaur, Shailesh Tripathi, CL Laxmipathi Gowda, GV Ranga Rao, HC Sharma, Suresh Pande and Mamta Sharma (2010). Chickpea Seed Production Manual. International Crops Research Institute for the Semi-Arid Tropics (ICRISAT) , India. 1-22pp.
Senthil G.;Williamson, B;Dinkins R.D.and Ramsay, G. (2004) An efficient transformation system for chickpea (Cicer arietinum L. ). Plant Cell Report . 22:576-583.
Sharmin R. A.; Akter, J ; Sarker, R. H; Hoque, M. I (2012). Agrobacterium- me diated Genetic Transformation of Local Cultivars of Chickpea (Cicer arietinum L.) Plant Tissue Cult. & Biotech. 22(1): 41-50.
Singh, KB. ; Malhotra, RS.; Halila, MH.; Knights, EJ. Verma, MM. (1994) Current status and future strategy in breeding chickpea for resistance to biotic and abiotic stress. Euphytica. 73: 137-149.
Tewari – Singh, N.; Sen, J.; Kiesecker, H. ; Reddy, VS.; Jacobsen, HJ. and Guha – Mukherjee, S. (2004). Use of herbicide or lysine plus threonine for non – antibiotic selection of transgenic chickpea. Plant Cell Pep. 22: 576-83.
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