Effect of mutagens on regeneration and growth of in vitro grown epicotyl segments of rough lemon seedlings (Citrus jambhiri Lush.)
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Abstract
The present study on the effect of mutagens on regeneration and growth of in vitro grown epicotyls segments of rough lemon seedlings (Citrus jambhiri Lush.) was carried out during the years 2009- 2010 and 2010- 2011 in the Tissue Culture Laboratory, Department of Fruit Science, Punjab Agricultural University, Ludhiana. Developmental characteristics of the in vitro grown epicotyls segments on regeneration media were treated employing gamma radiation in Gray(Gy) at 0, 5,10,15, 20,25,30,35,40 and 45 Gy; the alkylating agent ethyl methane sulfonate (EMS) and methyl methane sulfonate (MMS) each at 0, 0.1, 0.2, 0.3, 0.4,0.5 and 0.6% (v/v) were evaluated. Epicotyl segments from one month old in vitro grown seedling were cultured in regeneration medium (MS+BAP 1.0 mglit-1) under controlled laboratory conditions (25±2? C, 16 hr photoperiod, 2000 lux light). LD50, the dose required to kill half of the tested population corresponded to 35Gy for gamma radiation, 0.3% each for EMS and MMS treatments. Number of days taken for regeneration increased with increasing dose of gamma irradiation, EMS and MMS. Percent regeneration, number of buds, number of shoots, shoot length, number of leaves, internodal length, primary root length and number of secondary roots decreased with increasing dose of gamma radiation , EMS and MMS. The study would be beneficial to induce desirable variations in plant growth characteristics of rough lemon by the use of mutagens treatment.
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Keywords
Ethyl methane sulfonate, Gamma rays, Germination, In vitro, Methyl methane sulfonate, Rough lemon
References
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Chaturvedi, H. C., Singh, S. K., Sharma, A. K. and Agnihotri, S. (2001). Citrus tissue culture employing vegetative explants. Indian Journal of Experimental Biology. 39:1080-1095.
Datta, S. K., Misra, P. and Mandal, A. K. A. (2005). In vitro mutagenesis– a quick method for establishment of solid mutant in chrysanthemum.Current Science 88 (1): 155–158.
Davis,F. S. and Albrigo. I. G. (1994). Citrus. CAB International, Wallingford, UK: 254.
Dhatt, A. S., Rattanpal, H. S., Khokhar, R. and Singh, S. N. (2000). Standardising LD50 dose of gamma rays and EMS in Kinnow seeds. Journal of Research Punjab Agricultural University. 37 (3-4): 194-197.
Gad, A. A. and El-Sawah.,M. H. (1985). Comparative study on the relative mutagenic effectiveness and efficiency of gamma rays and some alkylating agents in peas. Egyptian Journal of Genetics and Cytology. 14(2):213-220.
Garcia-Agustin, P. and Primo-Millo, E. (1995). Selection of a NaCl tolerant citrus plant. Plant Cell Report. 14: 314-318.
Gilchrist, E. J. and Haughn, G.W. (2005). Tilling without a plough: a new method with applications for reverse genetics. Current Opinion in Plant Biology. 8: 211-215.
Gulsen, O., Uzun, A., Pala, H., Canihos, E. and Kafa, G. (2007). Development of seedless and mal secco tolerant mutant lemons through budwood irradiation. Scientia Horticulturae. 112 (2): 184–190.
Gupta, S. C. and Sharma, K. D. (1994). Mutation induced variability in rice. Madras Agricultural Journal.81(1): 50-52.
Hearn, C. J. (1984). Development of seedless orange and grapefruit cultivars through seed irradiation. Journal of the American Society for Horticultural Science. 109 (2): 270-273.
Hensz, R. A. (1971). Star Ruby, a new red fleshed grapefruit with distinct tree characters. Journal of the Rio Grande Valley Horticultural Society. 14: 21-25.
Jain, S. M. (2005). In vitro mutagenesis for improving date palm (Phoenix dactylifera L.) . Emirates Journal of Food and Agriculture. 24:400-407.
Jander, G., Baerson, S. R., Hudak, J. A., Gonzalez, K. A., Gruys, K. J., Last, R. L.(2003). Ethyl methane sulfonate saturation mutagenesis in Arabidopsis to determine frequency of herbicide resistance. Plant Physiology.131: 139-146.
Jawaharlal, M.,Sambandamorthy, S. and Irulappan, I. (1992). Effect of gamma ray and EMS on seed germination and seedling growth in acidlime Citrus aurantifolia Swingle. South Indian Horticulture. 369: 332–336.
Kaur, S. and Rattanpal, H.S.(2010). Effect of mutagens on in vitro seed germination and growth of rough lemon (Citrus jambhiri Lush.) seedlings. Indian Journal of Agricultural Sciences. 80(9):773-776.
Kawamura, Y., Sadao Uchiyama, Yakio Saito. (1989). A half embryo test for identification of gamma irradiated grapefruit. Journal of Food Science. 54 (2): 379-382.
Kayim, M. and Koe, N. K. (2006). The effects of some carbohydrates on growth and somatic embryogenesis in citrus callus culture. Scientia Horticulture. 109: 29-34.
Kerkadze, I. G. (1985). Induced mutations in subtropical crops. V. Biological and genetic effect of treating citrus with gamma-radiation. Subtrop Kult. 4: 104-110.
Khan, M. R., Qureshi. A. S., Syed, A. H. and Ibrahim. M. (2005). Genetic variability induced by gamma irradiation and its modulation with gibbrellic acid in M2 generation of Chickpea (Cicer arietinum L.). Pakistan Journal of Botany.37(2):285-292.
Khokhar, R. (1998). ‘Studies on mutagen induced variability in Kinnow mandarin.’ MSc. thesis. Punjab Agricultural University, Ludhiana.
Kim, Y., Schumaker, K. S. and Zhu, J. K.(2006). EMS mutagenesis of Arabidopsis. Methods in Molecular Biology .323: 101-103.
Kumar, A. and Mishra, M. N. (2004). Effect of gamma rays, EMS and NMU on germination, seedling vigour, pollen viability and plant survival in M1 and M2 generation of Okra ( Abelmoschus esculentus L. Moench.). Advances in Plant Science. 17(1): 295-297.
Latado, R. R., TulmannNeto, A., Ando A Iemma, A. F., Pompeu Junior, J., Figueiredo, J. O., Pio, R. M., Machado, M. A., Namekata, T., Ceravolo, L. and Rossi, A. C. (2001). Sweet orange ‘Pera’ mutants with low number of seeds obtained through mutation induction. RevistaBrasileira de Fruticultura. 23 (2): 339–344.
Latado, R. R., Tulmann Neto, A. T., Pompeu Junior, J., Figueiredo, J. O., Pio, R. M., Machado, M. A., Namekata, T., Ceravolo, L., Montes, S. M. N. M. and Rossi, A. C. (2006). Seedless and citrus canker tolerant mutant clones in sweet orange induced by gamma rays. Plant Mutation Reports. 1 (2): 21–22.
Legave, J. M., Tissue-Agostini, D. and Jacquemond, C. (1989). Physiological effects induced by acute gammairradiation of clementine, Citrus reticulata Blanco budsticks. Fruits Paris. 446: 329-333.
Ling, A. P. K., Chia, J. Y., Hussein, S. and Harun, A. R. (2008). Physiological responses of Citrus sinensis to gamma irradiation. World Applied Sciences Journal. 5 (1):12-19.
MacKenzie, J. L., Saade, F. E., Le, Q. H., Bureau, T. E., Schoen, D. J .(2005). Genomic mutation in lines of Arabidopsis thaliana exposed to ultraviolet-B radiation. Genetics. 171: 715-723.
Majeed, A., Khan, A. U. R., Ahmad, H. and Muhammad, Z. (2010). Gamma irradiation effects on some growth parameters of Lepidium sativum L. ARPN Journal of Agricultural and Biological Science. 5 (1): 39–42.
Mallick, E. H., Hajar, N. G. and Bairagi, P. (1978). Physiological and biochemical mutagens, a priority to induced mutation. Indian Journal of Experimental Biology. 13: 187-188.
Murashige, T. and Skoog, F. (1962). A revised medium for rapid growth and bioassay with tobacco tissue culture. Plant Physiology.15: 473-497.
Raghami, M. and Ghazvini, R. F. (2005). Effects of gamma irradiation on embryogenic callus and embryogenesis from habituated calluses of lime (Citrus aurantifolia var. Mexican lime). Iranian Journal of Agricultural Sciences. 36 (2): 425–434.
Ribeiro, R. V. and Machado, E. C. (2007). Some aspects of citrus ecophysiology in sub-tropical climates: revisiting photosynthesis under natural conditions. Brazilian Journal of Plant Physiology. 19:393-411.
Saini, H.K. and Gill, M. I. S.(2009). Induction of mutation in Rough lemon (Citrus jambhiri Lush.) using gamma rays. Journal of Horticultural Sciences. 4(1):41-44.
Sharma, L. K., Kaushal, M., Gill, M. I. S. and Bali, S. K. (2013). Germination and survival of Citrus jambhiri seeds and epicotyls after treating with different mutagens under in vitro conditions. Middle East Journal of Scientific Research. 16(2):250-255.
Sharma, G. and Sharma, R. L. (1996). Radio-sensitivity determination of LD50 in different apple cultivars. Haryana Journal of Horticultural Sciences. 251: 1-4.
Singh, S., Bansal, M. L., Singh, T. P. and Kumar, P.(1998). Statistical Methods for Research Workers. Kalyani Publishers, New Delhi.
Singh, R. R. and Singh, J. (1989). Genotypic variability relationship with EMS in a Trigonella species complex. Indian Journal of Horticulture. 461: 111-114.
Sparrow, A. H. and Gunckel, J. E. (1956). The effect of plants on chronic exposure to gamma radiation from radio Cobalt. Proceedings of International Conference on Peaceful Uses of Atomic Energy. Pp. 52-59.
Swaminathan, M. S. (1965). A comparison of mutation induction in diploids and polyploids. In: The use of mutations in plant breeding. Radiation Botany (Supplement). 5: 619-641.
Swaminathan, M. S., Chopra, V. L. and Bhaskaran, S. (1962). Chromosome aberrations and the frequency and spectrum of mutations induced by ethyl methane sulfonate in barley and wheat. Indian Journal of Genetics. 22: 192-207.
Thapa, C. B.(2004). Effect of acute exposure of gamma rays on seed germination and seedling growth of Pinus kesiya Gord and Pinus wallichiana A.B. Jacks. Our nature. 2 :13-17.
Toker, C., Uzen, B., Canci, H. and Ceylan, F. O. (2005).Effect of gamma irradiation on the shoot length of Cicer seeds. Radiation Physics and Chemistry.73:365-367.
Wan, S. Y., Deng, Z., Deng, X. X., Ye, X. and Zhang, W. C. (1991). Advances made in in vitro mutation breeding in citrus. Proceedings of International Symposium on the Contribution of Plant Mutation Breeding to Crop Improvement, Vienna, Austria. 1: 263-269.
Veleminsky, J. ,Gichner, T. and Pokorny,V.(1975). Caffeine enhancement of alkalyting agent-induced injury in barley: its connection to DNA single-strand and their repair. Mutation Research. 28(1):79-86.
Waqar, A., Wasin, A., Farooqi and Sattar, S. Jr. (1992). Effect of gamma-irradiation on the morphology of Kinnow seedlings. Proceeding of the First International Seminar on Citriculture in Pakistan 2-5 December, 1992.
Whittwer, S H. (1971). Radiation induced mutations in crop plants. Outlook Agriculture. 6: 205-217.
Chaturvedi, H. C., Singh, S. K., Sharma, A. K. and Agnihotri, S. (2001). Citrus tissue culture employing vegetative explants. Indian Journal of Experimental Biology. 39:1080-1095.
Datta, S. K., Misra, P. and Mandal, A. K. A. (2005). In vitro mutagenesis– a quick method for establishment of solid mutant in chrysanthemum.Current Science 88 (1): 155–158.
Davis,F. S. and Albrigo. I. G. (1994). Citrus. CAB International, Wallingford, UK: 254.
Dhatt, A. S., Rattanpal, H. S., Khokhar, R. and Singh, S. N. (2000). Standardising LD50 dose of gamma rays and EMS in Kinnow seeds. Journal of Research Punjab Agricultural University. 37 (3-4): 194-197.
Gad, A. A. and El-Sawah.,M. H. (1985). Comparative study on the relative mutagenic effectiveness and efficiency of gamma rays and some alkylating agents in peas. Egyptian Journal of Genetics and Cytology. 14(2):213-220.
Garcia-Agustin, P. and Primo-Millo, E. (1995). Selection of a NaCl tolerant citrus plant. Plant Cell Report. 14: 314-318.
Gilchrist, E. J. and Haughn, G.W. (2005). Tilling without a plough: a new method with applications for reverse genetics. Current Opinion in Plant Biology. 8: 211-215.
Gulsen, O., Uzun, A., Pala, H., Canihos, E. and Kafa, G. (2007). Development of seedless and mal secco tolerant mutant lemons through budwood irradiation. Scientia Horticulturae. 112 (2): 184–190.
Gupta, S. C. and Sharma, K. D. (1994). Mutation induced variability in rice. Madras Agricultural Journal.81(1): 50-52.
Hearn, C. J. (1984). Development of seedless orange and grapefruit cultivars through seed irradiation. Journal of the American Society for Horticultural Science. 109 (2): 270-273.
Hensz, R. A. (1971). Star Ruby, a new red fleshed grapefruit with distinct tree characters. Journal of the Rio Grande Valley Horticultural Society. 14: 21-25.
Jain, S. M. (2005). In vitro mutagenesis for improving date palm (Phoenix dactylifera L.) . Emirates Journal of Food and Agriculture. 24:400-407.
Jander, G., Baerson, S. R., Hudak, J. A., Gonzalez, K. A., Gruys, K. J., Last, R. L.(2003). Ethyl methane sulfonate saturation mutagenesis in Arabidopsis to determine frequency of herbicide resistance. Plant Physiology.131: 139-146.
Jawaharlal, M.,Sambandamorthy, S. and Irulappan, I. (1992). Effect of gamma ray and EMS on seed germination and seedling growth in acidlime Citrus aurantifolia Swingle. South Indian Horticulture. 369: 332–336.
Kaur, S. and Rattanpal, H.S.(2010). Effect of mutagens on in vitro seed germination and growth of rough lemon (Citrus jambhiri Lush.) seedlings. Indian Journal of Agricultural Sciences. 80(9):773-776.
Kawamura, Y., Sadao Uchiyama, Yakio Saito. (1989). A half embryo test for identification of gamma irradiated grapefruit. Journal of Food Science. 54 (2): 379-382.
Kayim, M. and Koe, N. K. (2006). The effects of some carbohydrates on growth and somatic embryogenesis in citrus callus culture. Scientia Horticulture. 109: 29-34.
Kerkadze, I. G. (1985). Induced mutations in subtropical crops. V. Biological and genetic effect of treating citrus with gamma-radiation. Subtrop Kult. 4: 104-110.
Khan, M. R., Qureshi. A. S., Syed, A. H. and Ibrahim. M. (2005). Genetic variability induced by gamma irradiation and its modulation with gibbrellic acid in M2 generation of Chickpea (Cicer arietinum L.). Pakistan Journal of Botany.37(2):285-292.
Khokhar, R. (1998). ‘Studies on mutagen induced variability in Kinnow mandarin.’ MSc. thesis. Punjab Agricultural University, Ludhiana.
Kim, Y., Schumaker, K. S. and Zhu, J. K.(2006). EMS mutagenesis of Arabidopsis. Methods in Molecular Biology .323: 101-103.
Kumar, A. and Mishra, M. N. (2004). Effect of gamma rays, EMS and NMU on germination, seedling vigour, pollen viability and plant survival in M1 and M2 generation of Okra ( Abelmoschus esculentus L. Moench.). Advances in Plant Science. 17(1): 295-297.
Latado, R. R., TulmannNeto, A., Ando A Iemma, A. F., Pompeu Junior, J., Figueiredo, J. O., Pio, R. M., Machado, M. A., Namekata, T., Ceravolo, L. and Rossi, A. C. (2001). Sweet orange ‘Pera’ mutants with low number of seeds obtained through mutation induction. RevistaBrasileira de Fruticultura. 23 (2): 339–344.
Latado, R. R., Tulmann Neto, A. T., Pompeu Junior, J., Figueiredo, J. O., Pio, R. M., Machado, M. A., Namekata, T., Ceravolo, L., Montes, S. M. N. M. and Rossi, A. C. (2006). Seedless and citrus canker tolerant mutant clones in sweet orange induced by gamma rays. Plant Mutation Reports. 1 (2): 21–22.
Legave, J. M., Tissue-Agostini, D. and Jacquemond, C. (1989). Physiological effects induced by acute gammairradiation of clementine, Citrus reticulata Blanco budsticks. Fruits Paris. 446: 329-333.
Ling, A. P. K., Chia, J. Y., Hussein, S. and Harun, A. R. (2008). Physiological responses of Citrus sinensis to gamma irradiation. World Applied Sciences Journal. 5 (1):12-19.
MacKenzie, J. L., Saade, F. E., Le, Q. H., Bureau, T. E., Schoen, D. J .(2005). Genomic mutation in lines of Arabidopsis thaliana exposed to ultraviolet-B radiation. Genetics. 171: 715-723.
Majeed, A., Khan, A. U. R., Ahmad, H. and Muhammad, Z. (2010). Gamma irradiation effects on some growth parameters of Lepidium sativum L. ARPN Journal of Agricultural and Biological Science. 5 (1): 39–42.
Mallick, E. H., Hajar, N. G. and Bairagi, P. (1978). Physiological and biochemical mutagens, a priority to induced mutation. Indian Journal of Experimental Biology. 13: 187-188.
Murashige, T. and Skoog, F. (1962). A revised medium for rapid growth and bioassay with tobacco tissue culture. Plant Physiology.15: 473-497.
Raghami, M. and Ghazvini, R. F. (2005). Effects of gamma irradiation on embryogenic callus and embryogenesis from habituated calluses of lime (Citrus aurantifolia var. Mexican lime). Iranian Journal of Agricultural Sciences. 36 (2): 425–434.
Ribeiro, R. V. and Machado, E. C. (2007). Some aspects of citrus ecophysiology in sub-tropical climates: revisiting photosynthesis under natural conditions. Brazilian Journal of Plant Physiology. 19:393-411.
Saini, H.K. and Gill, M. I. S.(2009). Induction of mutation in Rough lemon (Citrus jambhiri Lush.) using gamma rays. Journal of Horticultural Sciences. 4(1):41-44.
Sharma, L. K., Kaushal, M., Gill, M. I. S. and Bali, S. K. (2013). Germination and survival of Citrus jambhiri seeds and epicotyls after treating with different mutagens under in vitro conditions. Middle East Journal of Scientific Research. 16(2):250-255.
Sharma, G. and Sharma, R. L. (1996). Radio-sensitivity determination of LD50 in different apple cultivars. Haryana Journal of Horticultural Sciences. 251: 1-4.
Singh, S., Bansal, M. L., Singh, T. P. and Kumar, P.(1998). Statistical Methods for Research Workers. Kalyani Publishers, New Delhi.
Singh, R. R. and Singh, J. (1989). Genotypic variability relationship with EMS in a Trigonella species complex. Indian Journal of Horticulture. 461: 111-114.
Sparrow, A. H. and Gunckel, J. E. (1956). The effect of plants on chronic exposure to gamma radiation from radio Cobalt. Proceedings of International Conference on Peaceful Uses of Atomic Energy. Pp. 52-59.
Swaminathan, M. S. (1965). A comparison of mutation induction in diploids and polyploids. In: The use of mutations in plant breeding. Radiation Botany (Supplement). 5: 619-641.
Swaminathan, M. S., Chopra, V. L. and Bhaskaran, S. (1962). Chromosome aberrations and the frequency and spectrum of mutations induced by ethyl methane sulfonate in barley and wheat. Indian Journal of Genetics. 22: 192-207.
Thapa, C. B.(2004). Effect of acute exposure of gamma rays on seed germination and seedling growth of Pinus kesiya Gord and Pinus wallichiana A.B. Jacks. Our nature. 2 :13-17.
Toker, C., Uzen, B., Canci, H. and Ceylan, F. O. (2005).Effect of gamma irradiation on the shoot length of Cicer seeds. Radiation Physics and Chemistry.73:365-367.
Wan, S. Y., Deng, Z., Deng, X. X., Ye, X. and Zhang, W. C. (1991). Advances made in in vitro mutation breeding in citrus. Proceedings of International Symposium on the Contribution of Plant Mutation Breeding to Crop Improvement, Vienna, Austria. 1: 263-269.
Veleminsky, J. ,Gichner, T. and Pokorny,V.(1975). Caffeine enhancement of alkalyting agent-induced injury in barley: its connection to DNA single-strand and their repair. Mutation Research. 28(1):79-86.
Waqar, A., Wasin, A., Farooqi and Sattar, S. Jr. (1992). Effect of gamma-irradiation on the morphology of Kinnow seedlings. Proceeding of the First International Seminar on Citriculture in Pakistan 2-5 December, 1992.
Whittwer, S H. (1971). Radiation induced mutations in crop plants. Outlook Agriculture. 6: 205-217.
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Effect of mutagens on regeneration and growth of in vitro grown epicotyl segments of rough lemon seedlings (Citrus jambhiri Lush.). (2015). Journal of Applied and Natural Science, 7(1), 459-465. https://doi.org/10.31018/jans.v7i1.632
