Abstract
A cultivar of dessert banana, namely, Nanjanagudu Rasabale (NR), classified under group “silk” (of genotype AAB), is seriously under the threat of extinction due to its susceptibility to bacterial wilt and bunchy-top virus disease. A regeneration protocol using tissue culture method was developed (Venkatachalam et al. 2006), where a large number of plantlets were regenerated from leaf base explants. Simultaneously, a micropropagation protocol was also developed where high levels of up to 53.28 μM of benzylamino purine (BAP) and 55.80 μM of kinetin (Kn) were used. The progressive increase of cytokinins levels resulted in concomitant increase in shoot number, with a maximum of 80 shoot buds per segment in BAP (31.08 μM). The plantlets were analyzed for their genetic stability using randomly amplified polymorphic DNA (RAPD) and inter simple sequence repeats (ISSR) markers. A total of 50 RAPD and 12 ISSR primers resulted in 625 distinct and reproducible bands showing homogeneous RAPD and ISSR patterns. Band intensity histogram of each gel confirmed their monomorphic nature with no genetic variation among the plantlets analyzed. The present study has established for the first time that the regeneration and rapid micropropagation protocol developed through the present study will be of great use in conserving the endangered cultivar – NR – without risk of genetic instability.
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Bairu, M.W.; Fennell, C.W.; van Staden, J. The effect of plant growth regulators on somaclonal variation in Cavendish banana (Musa AAA cv. ‘Zelig’). Sci. Hort. 108:347–351; 2006
Bennici, A.; Anzidei, M.; Vendramin, G.G. Genetic stability and uniformity of Foeniculum vulgare Mill. regenerated plants through organogenesis and somatic embryogenesis. Plant Sci. 166:221–227; 2004
Bhagyalakshmi, N.; Singh, N.S. Role of liquid versus agar-gelled media in mass propagation and ex vitro survival in bananas. Plant Cell Tissue Organ Cult. 41:71–73; 1995
Blakesley, D.; Lenton, J.R. Cytokinin uptake and metabolism in relation to shoot multiplication in vitro. British Plant Growth Regulator Group Monograph. 16:87–99; 1987
Carvalho, L.C.; Goulao, L.; Oliveira, C.; Goncalves, J.C.; Amancio, S. RAPD assessment for identification of clonal identity and genetic stability of in vitro propagated chestnut hybrids. Plant Cell Tiss. Organ Cult. 77:23–27; 2004
Cronauer, S.S.; Krikorian, A.D. Somatic embryos from cultured tissues of triploid plantains (Musa ABB). Plant Cell Rep. 2:289–291; 1983
Damasco, O.P.; Graham, G.C.; Henry, R.J.; Adkins, S.W.; Smith, M.K.; Godwin, I.D. Random amplified polymorphic DNA (RAPD) detection of dwarf off-types in micropropagated Cavendish (Musa spp. AAA) bananas. Plant Cell Rep. 16:118–123; 1996
Dhed’a, D.; Dumortier, F.; Panis, B.; Vuylsteke, D.; De Langhe, E. Plant regeneration in cell suspension cultures of cooking banana ‘Bluggoe’ cultivar (Musa spp. ABB group). Fruits. 46:125–135; 1991
Goto, S.; Thakur, R.C.; Ishii, K. Determination of genetic stability in long-term micropropagated shoots of Pinus thunbergii Parl. using RAPD markers. Plant Cell Rep. 18:193–197; 1998
Israeli, Y.; Bassat, D.B.; Reuveni, O. Selection of stable clones which do not produce dwarf somaclonal variants during in vitro culture. Sci. Hort. 67:197–205; 1996
Lattoo, S.K.; Bamotra, S.; Sapru, D.R.; Khan, S.; Dhar, A.K. Rapid plant regeneration and analysis of genetic fidelity of in vitro derived plants of Chlorophytum arundinaceum Baker—an endangered medicinal herb. Plant Cell Rep. 25:499–506; 2006
Letham, D.S.; Gollnow, B.I. Regulators of cell division in plant tissues. Cytokinin metabolism in relation to radish cotyledon expansion and senescence. J. Plant Growth Reg. 7:129–145; 1985
Martin, K.P.; Pachathundikandi, S.; Zhang, C.L.; Slater, A.; Madassery, J. RAPD analysis of a variant of banana (Musa sp.) cv. grande naine and its propagation via shoot tip culture. In Vitro Cell. Dev. Biol. Plant. 42:188–192; 2006
Martins, M.; Sarmento, D.; Oliveira, M.M. Genetic stability of micropropagated almond plantlets as assessed by RAPD and ISSR markers. Plant Cell Rep. 23:492–496; 2004
Mendes, B.M.J.; Filippi, S.B.; Demetrio, C.G.B. A statistical approach to study the dynamics of micropropagation rates using banana (Musa spp.) as an example. Plant Cell Rep. 18:971; 1999
Murashige, T.; Skoog, F. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15:473–497; 1962
Novak, E.J.; Afza, R.; Van Duren, M.; Perea-Dallos, M.; Conger, B.V.; Xiaolang, T. Somatic embryogenesis and plant regeneration in suspension cultures of dessert (AA and AAA) and cooking bananas (Musa spp.). Bio-Technology. 7:154–159; 1989
Onguso, J.M.; Kahangi, E.M.; Ndiritu, D.W.; Mizutani, F. Genetic characterization of cultivated bananas and plantains in Kenya by RAPD markers. Sci. Hort. 99:9–20; 2004
Palombi, M.A.; Damiano, C. Comparison between RAPD and SSR molecular markers in detecting genetic variation in kiwifruit (Actinidia deliciosa A. Chev). Plant Cell Rep. 20:1061–1066; 2002
Peres, L.E.P.; Amar, S.; Kerbauy, G.B.; Salatino, A.; Zaffari, G.R.; Mercier, H. Effects of auxin, cytokinin and ethylene treatments on the endogenous ethylene and auxin-to-cytokinin ratio related to direct root tip conversion of Catasetum fimbriatum Lindl (Orchidaceae) into buds. J. Plant Physiol. 155:551–555; 1999
Pillay, M.; Ogundiwin, E.; Nwakanma, D.C.; Ude, G.; Tenkouano, A. Analysis of genetic diversity and relationships in East African banana germplasm. Theor. Appl. Genet. 102:965–970; 2001
Ramage, C.M.; Borda, A.M.; Hamill, S.D.; Smith, M.K. A simplified PCR test for early detection of dwarf off-types in micropropagated Cavendish Banana (Musa spp. AAA). Sci. Hort. 103:145–151; 2004
Ray, T.; Dutta, I.; Saha, P.; Das, S.; Roy, S.C. Genetic stability of three economically important micropropagated banana (Musa spp.) cultivars of lower Indo-Gangetic plains, as assessed by RAPD and ISSR markers. Plant Cell Tiss. Organ Cult. 85:11–21; 2006
Reuveni, O.; Israeli, Y. Measures to reduce somaclonal variation in in vitro propagated bananas. Acta Hort. 275:307–313; 1990
Ryynanen, L.; Aronen, T. Genome fidelity during short- and long-term tissue culture and differentially cryostored meristems of silver birch (Betula pendula). Plant Cell Tiss. Organ Cult. 83:21–32; 2005
Sahijram, L.; Soneji, J.R.; Bollamma, K.T. Analysing somaclonal variation in micropropagated bananas (Musa spp.). In Vitro Cell. Dev. Biol. Plant. 39:551–556; 2003
Sánchez, M.C.; Martínez, M.T.; Valladares, S.; Ferro, E.; Ana, M.; Viéitez. Maturation and germination of oak somatic embryos originated from leaf and stem explants: RAPD markers for genetic analysis of regenerants. J. Plant Physiol. 160: 699–707; 2003
Shenoy, V.B.; Vasil, I.K. Biochemical and molecular analysis of plants derived from embryogenic tissue cultures of napier grass (Pennisetum purpureum K. Schum). Theor. Appl. Genet. 83:947–955; 1992
Smith, M.K. A review of factors influencing the genetic stability of micropropagated banana. Fruits. 43:219–233; 1988
Turner, S.; Krauss, S.L.; Bunn, E.; Senaratna, T.; Dixon, K.; Tan, B.; Touchell, D. Genetic fidelity and viability of Anigozanthos viridis following tissue culture, cold storage and cryo-preservation. Plant Sci. 161:1099–1106; 2001
Ude, G.; Pillay, M.; Ogundiwin, E.; Tenkouano, A. Genetic diversity in an African plantain core collection using AFLP and RAPD markers. Theor. Appl. Genet. 107:248–255; 2003
Venkatachalam, L.; Sreedhar, R.V.; Bhagyalakshmi, N. Molecular analysis of genetic stability in long-term micropropagated shoots of banana using RAPD and ISSR markers. Electronic J. Biotechnol. DOI:10.2225/vol10-issue 1-fulltext-12, in press; 2007
Venkatachalam, L.; Thimmaraju, R.; Sreedhar, R.V.; Bhagyalakshmi, N. Direct shoot and cormlet regeneration from leaf explants of “silk” banana (AAB). In Vitro Cell. Dev. Biol. Plant. 42:262–269; 2006
Vuylsteke, D.R.; Swenne, R.; De Langhe, E.A. Somaclonal variation in plantains (Musa spp. AAB group) derived from shoot tip culture. Fruits. 46:429–439; 1991.
Wealth of India. Raw material, vol. 6 (L-M) New Delhi: CSIR; 1962
Williams, J.G.K.; Kubelik, A.R.; Livak, K.J.; Rafalski, J.A.; Tingey, S.V. DNA polymorphism amplified by arbitrary primers are useful as genetic marker. Nucleic Acids Res. 18:6531–6535; 1990
Zietjiewicz, E.; Rafalski, A.; Labuda, D. Genome fingerprinting by simple sequence repeat (SSR)-anchored polymerase chain reaction amplification. Genomics. 20:176–183; 1994
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The authors LV and RVS acknowledge the Council of Scientific and Industrial Research, India, Research Fellowship. Encouragement by Dr. G. A. Ravishankar, Head, Plant Cell Biotechnology Department (PCBT), and Dr. V. Prakash, Director, Central Food Technological Research Institute, for research activities are gratefully acknowledged.
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Venkatachalam, L., Sreedhar, R.V. & Bhagyalakshmi, N. Genetic analyses of micropropagated and regenerated plantlets of banana as assessed by RAPD and ISSR markers. In Vitro Cell.Dev.Biol.-Plant 43, 267–274 (2007). https://doi.org/10.1007/s11627-007-9028-7
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DOI: https://doi.org/10.1007/s11627-007-9028-7