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Prakash, D. P.
- Effect of Agrobacterium Infection Time, Co-Cultivation and Cell Density on in vitro Response in Hypocotyl of Eggplant (Solanum melongena L.)
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PDF Views:121
Authors
Affiliations
1 College of Horticulture, University of Horticultural Sciences, Munirabad - 583 233, Koppal, Karnataka, IN
2 Department of Biotechnology, Kuvempu University, Shankaraghatta, Shimoga - 577 451, IN
3 Division of Biotechnology, ICAR-Indian Institute of Horticultural Research, Hesaraghatta Lake Post, Bengaluru - 560 089, IN
1 College of Horticulture, University of Horticultural Sciences, Munirabad - 583 233, Koppal, Karnataka, IN
2 Department of Biotechnology, Kuvempu University, Shankaraghatta, Shimoga - 577 451, IN
3 Division of Biotechnology, ICAR-Indian Institute of Horticultural Research, Hesaraghatta Lake Post, Bengaluru - 560 089, IN
Source
Journal of Horticultural Sciences, Vol 11, No 1 (2016), Pagination: 21-26Abstract
The present study purports to assess the effect of Agrobacterium infection time, co-cultivation and cell density on in vitro response in hypocotyl explants of eggplant (brinjal) cv. Manjarigota. Agrobacterium (OD600 0.3-0.5) infection for 10-15 minutes (24.44±2.34%) was found to be optimum, while, higher or lower infection-time resulted in reduced callus initiation, shoot regeneration and explant survival. Explants with no (only Agrobacterium infection) or short (1 day) co-cultivation, showed reduced callus-initiation response and turned yellow, with no regeneration. Callus-initiation response increased from Day 1 (96.66±03.33%), and reached a maximum on Day 2 and Day 3 (100±00.00%). It decreased on further increase in co-cultivation time. Explants co-cultivated for three days showed highest regeneration response (30.00±02.96%) which thereafter reduced with further increase in co-cultivation time. Explants infected with Agrobacterium culture at 0.05 OD600 showed hardly any regeneration, and turned yellow and necrotic on the selection medium. Highest regeneration response (28.33±02.33%) was obtained in explants infected with 0.1 OD600 culture, and this gradually reduced as celldensity increased (upto 1.0 OD600), becoming zero in explants treated with cultures at 1.5 OD600 or above. Agrobacterium overgrowth was noticed on explants infected with cultures of 0.5 OD600 and above. Exposure of hypocotyl explants to higher cell-density, longer infection-time and prolonged co-cultivation regime resulted in severe necrosis of explants; time taken for development of Agrobacterium overgrowth was less with increase in the level of these factors. Regenerated shoots were healthy, green, elongated and showed ischolar_main induction on culture medium containing Kanamycin.Keywords
Eggplant, Manjarigota, Regeneration, Agrobacterium, PCR.References
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- Kumar, V.S. and Rajam, M.V. 2005. Enhanced induction of vir genes results in the improvement of Agrobacterium-mediated transformation of eggplant. J. Biochem. Biotech., 14:89-94
- Magioli, C. and Mansur, E. 2005. Eggplant (Solanum melongena L.): tissue culture, genetic transformation and use as an alternative model plant. Acta Bot. Bras., 19:139-148
- Magioli, C., Rocha, A.P.M., Pinheiro, M.M., Martins, S.G. and Mansur, E. 2000. Establishment of an efficient Agrobacterium-mediated transformation system for eggplant and study of a potential biotechnologically useful promoter. J. Pl. Biotech., 2:43-49
- Murashige, T. and Skoog, F. 1962. A revised medium for rapid growth and bioassays with tissue cultures. Physiol. Plantarum, 15:473-497
- Prakash, D.P., Deepali, B.S., Asokan, R., Ramachandra, Y.L., Lalitha Anand and Vageeshbabu S. Hanur. 2007a. Effect of explant type and growth regulators in the transformation of brinjal (Solanum melongena L.) cv. Manjarigota. J. Hortl. Sci., 2:94-98
- Prakash, D.P., Deepali, B.S., Asokan, R., Ramachandra, Y.L., Lalitha Anand and Vageeshbabu S. Hanur. 2007b. Effect of antibiotics and gelling agents in the transformation of brinjal (Solanum melongena L.) cv. Manjarigota. J. Hortl. Sci. 2:19-25
- Prakash, D.P., Deepali, B.S., Asokan, R., Ramachandra, Y.L., Shetti, D.L., Lalitha Anand and Vageeshbabu S. Hanur. 2008. Effect of growth regulators on in vitro complete plant regeneration in brinjal (Solanum melongena L.). Ind. J. Hort., 6:112-116
- Veluthambi, K., Gupta, K.A. and Sharma, A. 2003. The current status of plant transformation technologies. Curr. Sci., 84:368-379
- Zhang, Z., Coyne, D.P. and Mitra, A. 1997. Factors affecting Agrobacterium-mediated transformation of common bean. J. Amer. Soc. Hortl. Sci., 122:300-305
- Factors Affecting in Vitro Shoot Regeneration in Hypocotyls of Brinjal (Solanum melongena L.) in the Early Steps of Agrobacterium-Mediated Transformation
Abstract Views :235 |
PDF Views:114
Authors
Affiliations
1 College of Horticulture, Munirabad-583 233, IN
2 Department of Biotechnology, Kuvempu University, Shankaraghatta, Shimoga-577 451, IN
3 Division of Biotechnology, ICAR-Indian Institute of Horticultural Research, Hesaraghatta Lake Post, Bengaluru - 560 089, IN
1 College of Horticulture, Munirabad-583 233, IN
2 Department of Biotechnology, Kuvempu University, Shankaraghatta, Shimoga-577 451, IN
3 Division of Biotechnology, ICAR-Indian Institute of Horticultural Research, Hesaraghatta Lake Post, Bengaluru - 560 089, IN
Source
Journal of Horticultural Sciences, Vol 10, No 2 (2015), Pagination: 136-142Abstract
An attempt was made to assess the effect of size, age and position of the explant, pre-culture and high cytokinin concentration in the pre-culture medium on shoot regeneration in brinjal hypocotyls co-cultivated with Agrobacterium. The study was carried out using hypocotyl explants of brinjal cv. Manjarigota, Agrobacterium strain A208 and shoot regeneration medium (full-strength basal MS medium, 2μM BAP + 0.05μM NAA, 3% sucrose and 0.8% agar) containing Cefotaxime (250-500mg l-1) and Kanamycin (100mg l-1). Hypocotyl explants showed callus initiation and shoot regeneration response after 10-12 and 20-22 days of culture, respectively. Five-day-old explants did not survive Agrobacterium infection, and ten-day-old explants showed higher shoot regeneration (29±1.91%) than older explants. Explants of medium size (1cm long; 32±2.62%) from the apical region (38.57±2.61%) showed better shootregeneration ability than explants of any other size or region. A period of four days of pre-culture (33.33±3.76) was optimal best for best shoot-regeneration in hypocotyl explants. No regeneration was seen in hypocotyl explants at shorter or longer pre-culture period. High cytokinin (10μM) in shoot regeneration medium during pre-culture enhanced shoot regeneration response (47.27±2.98%) in explants co-cultivated with Agrobacterium. Effects of various factors documented in this study will be useful in developing an efficient Agrobacterium-mediated transformation protocol in brinjal cv. Manjarigota.Keywords
Eggplant, Pre-Culture on High BAP, Explant Characters, PCR.References
- Balappa, S.R. and Hugar, L.B. 2002. An economic evaluation of brinjal production and its marketing system in Karnataka. Agril. Market, 44:45-49
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- Chen, Q., Jelenkovic, G., Chin, C., Billings, S., Eberhardt, J. and Goffreda, J.C. 1995. Transfer and transcriptional expression of coleopteran Cry3B endotoxin gene of Bacillus thuringiensis in eggplant. J. Amer. Soc. Hort’l. Sci., 120:921-927
- Chy, Y.S. and Philips, G.C. 1987. High efficiency Agrobacterium-mediated transformation of Lycopersicon based on conditions favaourable for regeneration. Pl. Cell Rep., 6:105-108
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- Compton, M.E. 2000. Interaction between explant size and cultivar affects shoot organogenic competence of watermelon cotyledons. Hort’l. sci., 35:749-750
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- Fari, M., Nagy, I., Csányi, M., Mityko, J. and Rásfalvy, A. 1995. Agrobacterium-mediated genetic transformation and plant regeneration via organogenesis and somatic embryogenesis from cotyledon leaves in eggplant (Solanum melongena L. cv. ‘Kecskemeti lila’). Pl. Cell Rep., 15:82-86
- Frary, A. and Earle, E.D. 1996. An examination of factors affecting the efficiency of Agrobacterium-mediated transformation of tomato. Pl. Cell Rep., 16:235-240
- George, E.F. 1993. Plant propagation by tissue culture. Part 1. The technology. Exegenetics Ltd., Eddington, England
- Hamza, S. and Chupeau, Y. 1993. Re-evaluation of conditions for plant regeneration and Agrobacterium-mediated transformation from tomato. J. Expt’l. Bot., 44:18371845
- Hanur, V.S., Prakash, D.P., Deepali, B.S., Asokan, R., Ramachandra, Y.L., Riaz Mahmood and Lalitha Anand. 2006. Synergistic use of hypocotyl explants and high BAP pre-conditioning for enhanced transformation frequency in brinjal (Solanum melongena L.). J. Hort’l. Sci., 1:116-119
- Herath, P.S., Suzuki, T. and Hattori, K. 2005. Factors influencing Agrobacterium-mediated genetic transformation of Kenaf. Pl. Cell Tiss. Org. Cult., 82:201-206
- Kumar, V.S. and Rajam, M.V. 2005. Enhanced induction of vir genes results in the improvement of Agrobacterium-mediated transformation of eggplant. J. Biochem. Biotech., 14:89-94
- Lazzeri, P.A. and Dunwell, J.M. 1986. In vitro regeneration from seedling organs of Brassica oleracea var. italica Plenck. cv. Green Comet. II: Effect of light conditions and explant size. Annl. Bot., 58:699-710
- Ling, H., Kriseleit, D. and Ganal, M.W. 1998. Effect of ticarcillin/ potassium clavulanate on callus growth and shoot regeneration in Agrobacterium-mediated transformation of tomato. Pl. Cell Rep., 17:843-847
- Magioli, C. and Mansur, E. 2005. Eggplant (Solanum melongena L.): tissue culture, genetic transformation and use as an alternative model plant. Acta Bot. Brassilica, 19:139-148
- Magioli, C., Rocha, A.P.M., Pinheiro, M.M., Martins, S.G. and Mansur, E. 2000. Establishment of an efficient Agrobacterium-mediated transformation system for eggplant and study of a potential biotechnologically useful promoter. J. Pl. Biotech., 2:43-49
- Murashige, T. and Skoog, F. 1962. A revised method for rapid growth and bioassays with tissue cultures. Physiol. Pl., 15:473-497
- Nehra, N.S., Chibbar, R.N., Kartha, K.K., Datla, R.S.S., Crosby, W.L. and Stushnoff, C. 1990. Agrobacterium-mediated transformation of strawberry calli and recovery of transgenic plants. Pl. Cell Rep., 9:10-13
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- Prakash, D.P., Deepali, B.S., Asokan, R., Ramachandra, Y.L., Lalitha Anand and Vageeshbabu S. Hanur. 2007a. Effect of explant type and growth regulators in the transformation of brinjal (Solanum melongena L.) cv. Manjarigota. J. Hort’l. Sci., 2:94-98
- Prakash, D.P., Deepali, B.S., Asokan, R., Ramachandra, Y.L., Lalitha Anand and Vageeshbabu S. Hanur. 2007b. Effect of antibiotics and gelling agents in the transformation of brinjal (Solanum melongena L.) cv. Manjarigota. J. Hort’l. Sci., 2:19-25
- Prakash, D.P., Deepali, B.S., Asokan, R., Ramachandra, Y.L., Shetti, D.L., Lalitha Anand and Vageeshbabu S. Hanur. 2008. Effect of growth regulators on in vitro complete plant regeneration in brinjal (Solanum melongena L.). Ind. J. Hort., 64:112-116
- Sanyal, I., Singh, A.K., Kaushik, M. and Amla, D.V. 2005. Agrobacterium-mediated transformation of chickpea (Cicer arietinum L.) with Bacillus thuringiensis cry1Ac gene for resistance against pod borer insect Helicoverpa armigera. Pl. Sci., 168:1135-1146
- Sharma, P. and Rajam, M.V. 1995. Genotype, explant and position effects on organogenesis and embryogenesis in eggplant (Solanum melongena L.). J. Expt’l. Bot., 46:135-141
- Veluthambi, K., Gupta, K.A. and Sharma, A. 2003. The current status of plant transformation technologies. Curr. Sci., 84:368-379
- Effect of Age and Size of Hypocotyl Explant on in vitro Shoot Regeneration in Eggplant
Abstract Views :156 |
PDF Views:113
Authors
Affiliations
1 College of Horticulture, Munirabad-583 233, Koppal, UHS Bagalkot, Karnataka, IN
2 Division of Biotechnology, Indian Institute of Horticultural Research, Hesaraghatta Lake Post, Bangalore 560 089, IN
3 Department of Biotechnology, Kuvempu University, Shankaraghatta, Shimoga 577 451, IN
1 College of Horticulture, Munirabad-583 233, Koppal, UHS Bagalkot, Karnataka, IN
2 Division of Biotechnology, Indian Institute of Horticultural Research, Hesaraghatta Lake Post, Bangalore 560 089, IN
3 Department of Biotechnology, Kuvempu University, Shankaraghatta, Shimoga 577 451, IN
Source
Journal of Horticultural Sciences, Vol 7, No 2 (2012), Pagination: 203-205Abstract
In the present study, effect of size and age of hypocotyl explant on in vitro organogenetic responses was assessed in eggplant cv. Manjarigota. Size and age did not affect callus-initiation response, but showed marked influence on shoot regeneration response. Hypocotyl explants 1.5cm long showed highest shoot regeneration response (77.4%); either increase or decrease in size resulted in reduced response. Five to 15 day old hypocotyl explants showed direct shoot regeneration from cut ends, whereas 20-30 day old hypocotyl explants showed indirect shoot regeneration from callus produced on cut ends. Five day old explants were most responsive, with highest (91.23%) and thirty day old explants least responsive with reference to shoot regeneration response (20.85%). Shoot regeneration frequency decreased with increasing age, whereas shoot regeneration efficiency increased with increasing age of hypocotyl explants.Keywords
Eggplant, Shoot Regeneration, Age and Size, Hypocotyl Explants.- Evaluation of Annual Statice (Limonium sinuatum L.) Cultivars
Abstract Views :178 |
PDF Views:140
Authors
S. K. Nataraj
1,
P. M. Gangadharappa
1,
B. S. Reddy
1,
K. B. Naik
1,
S. J. Prashanth
1,
D. P. Prakash
1
Affiliations
1 Biotechnology Centre, P.B.-7648, Hulimavu, B.G. Road, Bangalore-560 076, IN
1 Biotechnology Centre, P.B.-7648, Hulimavu, B.G. Road, Bangalore-560 076, IN
Source
Journal of Horticultural Sciences, Vol 4, No 2 (2009), Pagination: 184-186Abstract
A field study was conducted during rabi season of 2003-2004 to evaluate the potential of statice cultivars as cut flower crop in the field of Department of Floriculture and Landscape Gardening, K.R.C. College of Horticulture, Arabhavi. The experiment was laid out as a Randomised Block Design with four replications. The panicles of each variety were harvested when calycys of individual flower have mostly opened and showing colour. The cultivar Turbo White recorded maximum plant height (21.78 cm), panicle length (80.94 cm), stem diameter (0.61 cm), no. of leaves (186.60), maximum fresh weight of panicles (530.09 g/plant), plant spread 62.15 cm2 and remained superior over others. The results are in agreement with the findings of Angadi (2000) in China aster. Maximum panicle production per plant (25.64) was recorded in cv. Turbo White, which was on par with the cultivar Turbo Carmine (22.54). The results were in line with the findings of Kumar et al. (1998) in annual statice Turbo White. The cultivar Turbo White was good in quality parameters by recording maximum panicle length, girth and number of branches per panicle. It was also vigorous in vegetative growth. Therefore, the cultivar could produce better quality panicles and found to be suitable for semi arid regions.Keywords
Evaluation, Annual Statice, Growth, Yield, Quality.- Effects of Growth Regulators and Explant-Type on Agrobacterium-Mediated Transformation in Brinjal (Solanum melongena L.) cv. Manjarigota
Abstract Views :208 |
PDF Views:108
Authors
D. P. Prakash
1,
B. S. Deepali
1,
R. Asokan
1,
Y. L. Ramachandra
2,
Lalitha Anand
1,
Vageeshbabu S. Hanur
1
Affiliations
1 Division of Biotechnology, Indian Institute of Horticultural Research, Hesaraghatta Lake Post, Bangalore - 560 089, IN
2 Department of Biotechnology, Kuvempu University, Shankaraghatta, Shimoga, IN
1 Division of Biotechnology, Indian Institute of Horticultural Research, Hesaraghatta Lake Post, Bangalore - 560 089, IN
2 Department of Biotechnology, Kuvempu University, Shankaraghatta, Shimoga, IN
Source
Journal of Horticultural Sciences, Vol 2, No 2 (2007), Pagination: 94-98Abstract
Effects of growth regulators and type of explants on transformation and in vitro morphogenetic responses of brinjal cv. Manjarigota were studied. Both hypocotyl and cotyledonary explants showed marked influence on in vitro morphogenetic responses after Agrobacterium co-cultivation. Hypocotyl explants showed callus initiation and regeneration responses earlier than cotyledonary leaves. Hypocotyl explants were found to be better than cotyledonary leaf explants in regenerating shoots after Agrobacterium co-cultivation. There was delay and reduction in both callus and regeneration responses in Agrobacterium co-cultivated explants. Hypocotyl explants showed the highest regeneration response on MS medium containing 2 μM BAP and 0.05 μM NAA while cotyledonary leaves did not show regeneration response after Agrobacterium co-cultivation. However, they showed green buds on MS medium containing 10 μM BAP and 1 μM NAA, which could not differentiate into shoots. Overall, hypocotyl explants were found better in regenerating shoots after Agrobacterium co-cultivation.Keywords
Growth Regulators, Explant, Brinjal, Transformation.- Effect of Antibiotics and Gelling Agents in Transformation of Brinjal (Solanum melongena L.) cv. Manjarigota
Abstract Views :226 |
PDF Views:112
Authors
D. P. Prakash
1,
B. S. Deepali
1,
R. Asokan
1,
Y. L. Ramachandra
2,
Lalitha Anand
1,
Vageeshbabu S. Hanur
1
Affiliations
1 Division of Biotechnology, Indian Institute of Horticultural Research, Hessaraghatta Lake Post, Bangalore – 560 089, IN
2 Department of Biotechnology, Kuvempu University, Shankaraghatta, Shimoga, IN
1 Division of Biotechnology, Indian Institute of Horticultural Research, Hessaraghatta Lake Post, Bangalore – 560 089, IN
2 Department of Biotechnology, Kuvempu University, Shankaraghatta, Shimoga, IN
Source
Journal of Horticultural Sciences, Vol 2, No 1 (2007), Pagination: 19-25Abstract
A study was conducted to find out the effect of antibiotics and gelling agents on Agrobacterium-mediated transformation using hypocotyl explants of brinjal cv. Manjarigota. Hypocotyl explants of brinjal were found to be sensitive even to the lowest level of kanamycin (25 mg/l) tested. Explants that showed increased callus initiation and regeneration response upon cocultivation with Agrobacterium and on kanamycin at 100 mg/l were selected as this indicated a highly effective selection pressure. Cefotaxime did not affect regeneration response and at 500 mg/l, it effectively inhibited Agrobacterium overgrowth completely on Agrobacterium cocultivated hypocotyl explants. There were marked differences in regeneration response in hypocotyl explants cultured on medium solidified with various gelling agents indicating the influence of gelling agent on the activity of kanamycin in culture medium, which indirectly affects selection and recovery of transformants. Antibiotics and gelling agents could therefore affect, directly or indirectly, transformation of brinjal cv. Manjarigota.Keywords
Solanum melongena, Kanamycin, Cefotaxime, Gelling Agents, Transformation.- Synergistic Use of Hypocotyl Explants and High Bap Preconditioning for Enhanced Transformation Frequency in Brinjal (Solanum melongena L.)
Abstract Views :214 |
PDF Views:112
Authors
Vageeshbabu S. Hanur
1,
D. P. Prakash
1,
B. S. Deepali
1,
R. Asokan
1,
Y. L. Ramachandra
2,
Riaz Mahmood
2,
Lalitha Anand
1
Affiliations
1 Division of Biotechnology, Indian Institute of Horticultural Research, Hessaraghatta Lake Post, Bangalore 560 089, IN
2 Department of Biotechnology, Kuvempu University, Shankaraghatta, Shimoga, IN
1 Division of Biotechnology, Indian Institute of Horticultural Research, Hessaraghatta Lake Post, Bangalore 560 089, IN
2 Department of Biotechnology, Kuvempu University, Shankaraghatta, Shimoga, IN