Combined Effect of Explant, Inoculation Time and Co-cultivation Period on Agrobacterium - Mediated Genetic Transformation in Cucumber (Var. Shital)
[1]
S. M. Faisal, Horticulture Division, Bangladesh Agricultural Research Institute, Agricultural Research Station, Khulshi, Pahartali, Chittagong, Bangladesh.
[2]
M. S. Haque, Department of Biotechnology, Faculty of Agriculture, Bangladesh Agricultural University, Mymensingh, Bangladesh.
[3]
K. M. Nasiruddin, Department of Biotechnology, Faculty of Agriculture, Bangladesh Agricultural University, Mymensingh, Bangladesh.
Combined effect of explant, inoculation time and co-cultivation periodon Agrobacterium-mediated genetic transformation in cucumber was observed. Transformation ability was examined by histochemical assay of GUS reporter gene in survived calli. Conspicuous GUS positive (blue colour) region were detected in callus tissue. There were 3 factors in this investigation. Factor A consisted of three types of explants viz. leaf, nodal and internodal callus, factor B consisted of two durations of inoculation time viz. 3 and 5 min and factor C consisted of two co-cultivation periods viz. 24 and 48 hours. The highest number (4.04) and percentage (67.38%) of GUS +vecalliwas obtained from leaf explants when they were immersed in Agrobacterium suspension for 5 min and kept into co-cultivation media for 48 hrs.The lowest number(0.95) and percentage (15.89%) of GUS +ve calluswere observed in internodal explants when they were inoculated in Agrobacterium for 3 min and then co-cultivated for 24 hrs.
Cucumber, Genetic Transformation, Abiotic Stress
[1]
BBS. Statistical pocket book of Bangladesh. Bangladesh Bureau of statistics, Ministry of Planning, Government of the People’s Republic of Bangladesh, Dhaka, Bangladesh. 2008, pp. 209.
[2]
BBS. Yearbook of Agricultural Statistics of Bangladesh. Bangladesh Bureau of statistics, Ministry of Planning, Government of the People’s Republic of Bangladesh, Dhaka, Bangladesh. November. 2005, pp. 34, 97.
[3]
BBS. Monthly statistical bulletin-Bangladesh. Bangladesh Bureau of statistics, Ministry of Planning, Government of the People’s Republic of Bangladesh, Dhaka, Bangladesh. June. 2006, pp. 55.
[4]
S. N. Begam. Agrobacterium-mediated transformation of potato for abiotic stress tolerance. MS thesis. Dept. of Biotechnology. Bangladesh Agricultural University, Mymensingh. 2007, pp. 17-50.
[5]
H. S. Chawla. Introduction to Plant Biotechnology. Oxford & IBH publishing Co. Pvt. Ltd., 66 Janapath, New Delhi 110001, India. 2002, pp. 40-42.
[6]
R. C. Gardner. Gene transfer into tropical and subtropical crops. Scientia Hort. 1993, pp. 55, 65-82.
[7]
R. A. Jefferson, T. A. Kavanagh and M. W. Bevan. GUS Fusion: β- glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J., 6. 1986, pp 3901-3907.
[8]
S. Mahajan and N. Tuteja. Direct submission J. plant molecular Biol. I. C. G. E. B., New Delhi 110067, India, 2005.
[9]
T. Murashige and F. Skoog. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant, 15. 1962, pp. 473-497.
[10]
S. Nishibayashi, H. Kaneko and T. Hayakawa. Transformation of cucumber (Cucumissativus L.) plants using Agrobacterium tumefaciensand regeneration from hypocotyl explants. Plant Cell Reports, 15. 1996, pp. 809-814.
[11]
I. L. Nonnecki. Vegetable Production. An AVI book, Van Nostrand Reinhold, 115 Fifth Avenue, New York 10003, USA, 1989, pp. 509-526.
[12]
L. C. Peirce. Vegetables: characteristics, production and marketing. John Wiley and Sons, USA. 1987, pp357-366.
[13]
P. A. Rajagopalan and R. Perl-Treves. Improved cucumber transformation by a modified explant dissection and selection protocol. Hort. Science, 40 (2). 2005, pp. 431-435.
[14]
M. Roy, R. K. Jain, J. S. Rohila and R. Wu. Production of agronomically superior transgenic rice plants using Agrobacterium transformation methods: Present status and future perspectives. Current Science, Vol. 79, No. 7. 2000, pp 954-959.
[15]
W. B. Wang and A. Altman. Plant responses to drought, salinity and extreme temperatures: towards genetic engineering for stress tolerance. Intl. J. Plant. Biol., 218. 2003. pp 1-14.
[16]
Mohiuddin, A. K. M., K. Harikrishna, M. K. U. Chowdhury, Z. C. Abdullah and S. Napis. Influence of acetosyringone on Agrobacterium – mediated of cucumber (Cucumissativus L.). Plant Tissue Cult. 10(2). 2000, pp 167-173.
[17]
Vengadesan, G., P. Anand, N. Selvaraj, R. Perl-Treves and A. Ganapathi. Transfer and expression of nptii and bar genes in cucumber (Cucumissativus L.). In vitro cell. dev. Biol. 602. 2004. pp. 1-6.
[18]
Raharjo, S. H. T., M. O. Hernadez, Y. Y. Zhang and Z. K. Punja. Transformation of pickling cucumber with chitinase-encoding genes using Agrobacterium tumefaciens. Plant Cell Reports. 15. 1996, pp 591-596.
[19]
Chee, P. P. Transformation of Cucumissativus L. tissue by Agrobacterium tumefaciensand the regeneration of transformed plants. Plant Cell Reports. 9. 1990, pp 245-248.
[20]
Chee, P. P. and J. L. Slightom. Transfer and expression of cucumber mosaic virus coat protein gene in the genome of Cucumissativus. J. Amer. Soc. Hort. Sci. 116 (6). 1991, pp 1098-1102.
[21]
Trulson, A.J., R.B. Simpson and E.A. Shahin. Transformation of cucumber (Cucumissativus L.) plants with Agrobacterium rhizogenes. Theor Appl Genet. 73. 1986, pp 11-15.
[22]
Sarker, R.H., A. Biswas, B. M. Mustafa, S. Mahbub and M.I. Hoque. Agrobacterium-mediated transformation of lentil (Lens culinaris Medik.). Plant Tissue Cult. 13(1). 2003, pp 1-12.
[23]
Rachmawati, D. and H. Anzai. Studies on the callus induction, plant regeneration and transformation of Javanica rice cultivars. Plant Biotechnology. 23. 2006, pp 521-524.
[24]
Ding, L.C., C.Y. Hu, K.W. Yeh and P.J. Wang. 1998. Development of insect resistant transgenic cauliflower plants expressing the trypsin inhibitor gene isolated from local sweet potato. Plant Cell Rep., 17(11): 854-860.
[25]
Tsukazaki, H., Y. Kuginuki, R. Aida and T. Suzuki. 2002. Agrobacterium tumefaciens-mediated transformation of a doubled haploid line of cabbage. Plant Cell Rep. 21(3): 257-262.
[26]
Zhang, F. L., Y. Takahata, M. Watanabe and J. B. Xu. 2000. Agrobacterium-mediated transformation of cytoledonaryexpalnts of Chinese cabbage (Brassica campestris L. spp. pekinensis). Plant Cell Rep. 19: 569-575.
[27]
Belarmino, M.M. and M. Mii. 2000. Agrobacterium-madiated genetic transformation of a phalaenopsis orchid. Plant Cell Rep. 19: 435-442.
[28]
Yen, B., M. S. S. Reddy, G. B. Collins and R. D. Dinkins, 2000. Agrobacterium tumefaciens mediated transformation of soybean [Glycine max (L.) merriell] using immature zygotic cotyledon explants. Plant Cell Rep. 19(11): 1090-1097.
