Seasonal variation in contamination and browning of Acacia nilotica nodal explants in vitro
Article Main
Abstract
Major problems encountered in establishing axenic cultures are contamination and browning of explants and media. Contamination is initiated in explants excised from soil-borne tissues or adult trees. Several sterilizing agents (Tween 20, chlorine water, silver nitrate, mercuric chloride, etc.) or a wash in antibiotic solution are recommended to reduce this. . Establishing in vitro cultures of Acacia nilotica old tree nodal explants was a serious problem. The present study was undertaken to create a suitable protocol for in vitro micropropagation of nodal explants of Acacia nilotica subsp. indica. Contamination was controlled by sterilizing the explants by washing in polysan (5%, v/v along with 500mg/l PVP), thorough washing under tap water, 70% alcohol treatment and 0.1% HgCl2 treatment. Plant tissues release phenolic substances through their cut ends, which turn the media dark brown and toxic. Addition of various antioxidants or/and transfer of explants to fresh medium twice or thrice, at a few days’ intervals may overcome the problem. Collection of explants in antioxidant solution and a wash in antioxidant solution prior to inoculation was helpful in reducing the phenolic exudation in the present investigations. The addition of antioxidant (citric acid) to the medium also checked browning to some extent. The rate of infection and browning of explants varied in different seasons, the maximum being during winters and the minimum during summers of 2019. This was inversely related to the morphogenic response of explants i.e. maximum caulogenesis occurred in vitro during July (6.3±1.4 shoots per explant). The in vitro raised shoots showed 100% rooting on 2mg/l IBA augmented Nitsch’s (N) medium.
Article Details
Article Details
Keywords
Boric acid (H3BO3), Mercuric chloride (HgCl2), Polyvinylpyruvic acid (PVP), Seasonal variation
References
Ahmad, I., Jaskani, M. J., Nafees, M., Ashraf, I. & Qures, R. (2016). Control of media browning in micro propagation of guava (Psidium guajava l.). Pakistan Journal of Botany, 48(2), 713-716.
Alagarsamy K, Shamala L.F. & Wei S. (2018). Influence of media supplements on inhibition of oxidative browning and bacterial endophytes of Camellia sinensis var. sinensis. 3 Biotech; 8, 1-7. https://doi.org/10.1007/s13205-018-1378-9.
Amente, G. & Chimdessa, E. (2021). Control of browning in plant tissue culture: A review. Journal of Scientific Agriculture, 5, 67-71. https://doi.org/10.25081/jsa.2021.v5.7266.
Babaei, N., Abdullah, N.A., Saleh, G. & Abdullah, T. (2013). Control of contamination and explant browning in Curculigo latifolia in vitro cultures. Journal of Medicinal Plants, 7, 448-454. https://doi.org/10.5897/JMPR012.859.
Basto S., Serrano C. & de Jaramillo E.H. (2012). Effects of donor plant age and explants on in vitro culture of Cedrela montana Moritz ex Turcz. Universitas Scientiarum, 17 (3), 263-271. http://www.scielo.org.co/scielo.php?script=sci_arttext&pid=S0122-74832012000300002&lng=en&nrm=iso
Beckman, C. H. (2000). Phenolic-storing cells: keys to programmed cell death and periderm formation in wilt disease resistance and in general defence responses in plants? Physiological and Molecular Plant Pathology, 57(3), 101–110. https://doi.org/10.1006/ pmpp.2000.0287
Benisheikh A.A.G., Tijani Y., Askira N.K., Waziri M.S. & Awana A.U. (2015). Prevalence and management of laboratory microbial contaminants in plant tissue culture. International Journal of Research, 2(6), 625-630.
Bhat, S.N.; Khalil, A.; Nazir, N.; Mir, M.A.; Khan, I.; Mubashir, S.S.; Dar, M.S.; Wani, S.H.; Hossain, M.A. (2022) In Vitro Prevention of Browning in Persian Walnut (Juglans regia L.) cv. Sulaiman. Int. J. Plant Biol., 13, 330-342. https://doi.org/10.3390/ijpb13030027
Cai, X., Wei, H., Liu, C., Ren, X., Thi, L. T., & Jeong, B. R. (2020). Synergistic Effect of NaCl Pretreatment and PVP on browning suppression and callus induction from petal explants of Paeonia lactiflora Pall. ‘Festival Maxima’. Plants (Basel, Switzerland), 9(3), 346. https://doi. org/10.3390/plants9030346
Chai, J., Gao, Y., Dong, Y., Kong, L., & Zhang, Y. (2018). Browning Treatment in Tissue Culture of ‘Hongyang’ Kiwifruit. IOP Conference Series Materials Science and Engineering, 452(2): 022075. https://doi.org/10.1088/1757-899X/452/2/022075
Devi S. & Yadav N.R. (2022) Role of explants collecting seasons and sterilization process on in vitro regeneration of guava: A technology hurdle. The Pharma Innovation Journal, 11(5): 1244-1248.
Dewan, A.; Nanda, K. & Gupta, S.C. (1992). In vitro micropropagation of Acacia nilotica subsp.indica Brenan via cotyledonary nodes. Pl. Cell Rep. 12: 18-21 https://doi.org/10.1007/BF00232415.
Dobránszki, J., & Teixeira da Silva, J. A. (2010). Micropropagation of apple—A review. Biotechnology Advances, 28(4), 462–488. https://doi.org/10.1016/j.biotechadv.20 10.02.008
Gioushy, S.F.E.; Liu, R.; Fan, H.K. A complete protocol to reduce browning during coconut (cocos nucifera L.) tissue culture through shoot tips and inflorescence explants. (2020) Plant Arch. 20, 2196–2204.
Huang L., Wang H., Shahid M.Q. & Zeng B. (2019). Chlorothalonil: an effective bacteriostatic agent for bud induction of Acacia auriculiformis under open condition (non-axenic). Plant Methods. 15:5. https://doi.org/10.1186/s13007-019-0390-3.
Irshad, M., Rizwan, H. M., Debnath, B., Anwar, M., Li, M., Liu, S., He, B., & Qiu, D. (2018). Ascorbic acid controls lethal browning and pluronic f-68 promotes high-frequency multiple shoot regeneration from cotyldonary node explant of okra (Abelmoschus esculentus L.). Hortscience, 53(1), 183–190. https://doi.org/10.21273/HORTSCI12315-18
Jones, A. M., & Saxena, P. K. (2013). Inhibition of phenylpropanoid biosynthesis in Artemisia annua L.: a novel approach to reduce oxidative browning in plant tissue culture. PloS one, 8(10): 76802. https://doi.org/10.1371/journal.pone.0076802
Krishna, H., Sairam, R. K., Singh, S. K., Patel, V. B., Sharma, R. R., Grover, M., Nain, L., & Sachdev, A. (2008). Mango explants browning: Effect of ontogenic age, mycorrhization and pre-treatments. Scientia Horticulturae, 118(2), 132-138. https://doi.org/10.1016/j.scienta.2008.05.040
Kuppusamy, S. Seenivasan R., Subramanian S., Chinnaraj, S., Rajasree S., Kathirvel, B., Thamaraiselvi, K. (2019) Optimizing the sterilization methods for initiation of the five different clones of the Eucalyptus hybrid species, Biocatalysis and Agricultural Biotechnology, Volume 22,101361, ISSN 1878-8181, https://doi.org/10.1016/j.bcab.2019.101361.
Martini, A. N., Papafotiou, M. & Vemmos, S. N. (2013). Season and Explant Origin Affect Phenolic Content, Browning of Explants, and Micropropagation of ×Malosorbus florentina (Zucc.) Browicz, HortScience horts, 48(1), 102-107. https://doi.org/10.21273/HORTSCI.48.1.102
Mohammadi, M., Bagheri, A., Marashi, H., Moshtaghi, N., & Balandari, A. (2011). Investigation into Seasonal Effect and Browning Inhibitor on Callus Regeneration of Seedless Barberry (Berberis vulgaris var. asperma). Plant Tissue Culture and Biotechnology, 21(2), 161–168. https://doi.org/10.3329/ptcb.v21i2.10239
Mudoi K.D., Saikia S.P. & Borthakur, M. (2014). Effect of nodal positions, seasonal variations, shoot clump and growth regulators on micropropagation of commercially important bamboo, Bambusa nutans Wall. ex. Munro. African Journal of Biotechnology, 13, 1961-1972. https://doi.org/10.5897/AJB2014.13659
Narang A. (2022). In vitro micropropagation of Acacia holosericea ex. G.Don through cotyledons. International Journal of Multidisciplinary Educational Research, 11(1,7), 70-75. http://ijmer.in.doi./2022/11.01.135
Ndakidemi, C. F., Mneney, E., & Ndakidemi, P. A. (2014). Effects of ascorbic acid in controlling lethal browning in in vitro culture of Brahylaena huillensis using nodal segments. American Journal of Plant Sciences 5(1), 187–191. https://doi.org/10.4236/ajps.2014.51024
Omamor I, Asemota, a, Eke, C, Eziashi, E. (2007). Fungal contaminants of the oil palm tissue culture in Nigerian institute for oil palm research (NIFOR) African Journal of Agricultural Research 2(10), 534-537.
Permadi, N., Nurzaman,M., Alhasnawi, A.N., Doni, F. & Julaeha, E.(2023). Managing Lethal Browning and Microbial Contamination in Musa spp. Tissue Culture: Synthesis and Perspectives. Horticulturae, 9(4), 453. https://doi.org/10.3390/horticulturae9040453
Sarkar P.K., Sinha A., Bhatt B.P., Kumar R., Sarkar A., Banerjee S., Ghosh J., Kujur R.S. & Kulkarni N. (2019) Effect of pretreatment of explants and in vitro culture media in controlling infection and browning in Kusum [Schleichera oleosa (Lour.) Oken] IJCS, SP6, 914-917. http://krishi.icar.gov.in/jspui/handle/123456789/53533
Selvarajan, E., Veena, R., & Kumar, N. M. (2008). Polyphenol oxidase, beyond enzyme browning, a review. Microbial Bioprospecting for Sustainable Development (pp. 203-222), Singapore, Springer
Shimelis, D., Bantte, K., & Feyissa, T. (2015). Effects of Polyvinyl Pyrrolidone and Activated Charcoal to Control Effect of Phenolic Oxidation on In Vitro Culture Establishment Stage of Micropropagation of Sugarcane (Saccharum Officinarum L.). Advances in Crop Science and Technology, 3(4), 184. https://doi.org/10.4172/2329-8863.1000184
Shukla, A., Narang A. & Kaur S. (2023). Effect of Adenine Sulphate on In Vitro Micropropagation of Acacia holosericea A. Cunn. ex G. Don Leaflets and Leaf Rachis Explants. Journal of Tropical Life Science, 13(3), 497–502. https://doi.org/10.11594/jtls.13.03.08.
Silva G.M., Xavier M.A., da Silva P.A., Saldanha C.W. & Stein V.C. (2020). Microbial contamination in micropropagation: causes and prevention. Plant Cell, Tissue and Organ Culture, 142(2), 215-227. https://doi.org/10.1007/s11240-020-01819-6.
Singh V., Ankur T., Chauhan P.K., Kumari, P. & Kaushal, S. (2011). Identification and prevention of bacterial contamination on explants used in plant tissue culture labs. International Journal of Pharmacy and Pharmaceutical Sciences, 3(4), 160-163.
Singh, N. V., Singh, S. K. & Patel, V. B. (2011). In vitro culture establishment studies on pomegranate. Indian Journal of Horticulture, 68(3), 307-311.
Singh, P., & Patel, R. M. (2016). Factors affecting in vitro degree of browning and culture establishment of pomegranate. African Journal of Plant Science, 10(2), 43-49. https://doi.org/10.5897/AJPS2013.1119
Sivanesan, I., Muthu, M., Gopal, J., Tasneem, S., Kim, D.H., Oh & J. W. (2021) A Fumigation-Based Surface Sterilization Approach for Plant Tissue Culture. Int J Environ Res Public Health, 18(5), 2282. https://doi.org/10.3390/ijerph18052282. PMID: 33668989; PMCID: PMC7967719.
Sultana J, Rahman M.M., Karim R., Khan R.H., & Hakim M.A. (2021). Seasonal variation in phytochemical constituents and antioxidant activity of Musa acuminata cv. Barjahaji: an approach towards micropropagation. Bangladesh Journal of Botany, 50(2), 291-300. https://doi.org/10.3329/bjb.v50i2.54100.
Tao, Fj., Zhang, Zy., Zhou, J. & Yao, N. & Wang, Dm. (2007). Contamination and browning in tissue culture of Platanus occidentalis L. Forestry Studies in China, 9: 279-282. https://doi.org/10.1007/s11632-007-0044-9.
Titov, S., Bhowmik, S. K., Mandal, A., Alam, M. S., & Uddin S. N. (2006). Control of phenolic compound secretion and effect of growth regulators for organ formation from Musa spp. cv. Kanthali floral bud explants. American Journal of Biochemistry and Biotechnology, 2(3), 97–104. https://doi.org/10.3844/ajbbsp.2006.97
TheodosyMsgoya, Helen Kanyaghu, Josiah Mutigitu, MatesoKulebelwa (2012). Identification and management of microbial contaminants of banana in vitro cultures, Tanzania, 55:3987-3994
Varghese, N. & Joy, P.P. (2016). Plant tissue culture contaminants identification and its response to fumigation. https://www.researchgate.net/publication/3 06034945
Yahia H A Elbasheer., Alem A A., Osman E I E (2019). Feasible Surface Sterilization Protocol for Microbial Contamination of Some Forest Trees on in vitro Cultures Explants. Acta Scientific Microbiology 2.5: 55-60.
Wang N., Yang C., Pan Z., Liu Y. & Peng S. (2015). Boron deficiency in woody plants: various responses and tolerance mechanisms. Front Plant Sci. 2, 916. https://doi.org/10.3389/fpls.2015.00916.
Zafar S., Abbasi B.H., & Aqil M. (2017). Influence of seasonal variation on the production of bioactive compounds and antioxidant activity in cell cultures of Cichorium intybus L. and Cynara cardunculus L. Journal of Applied Research on Medicinal and Aromatic Plants, 4, 9-18. https://doi.org/10.1016/j.jarmap.2016.09.002.
Alagarsamy K, Shamala L.F. & Wei S. (2018). Influence of media supplements on inhibition of oxidative browning and bacterial endophytes of Camellia sinensis var. sinensis. 3 Biotech; 8, 1-7. https://doi.org/10.1007/s13205-018-1378-9.
Amente, G. & Chimdessa, E. (2021). Control of browning in plant tissue culture: A review. Journal of Scientific Agriculture, 5, 67-71. https://doi.org/10.25081/jsa.2021.v5.7266.
Babaei, N., Abdullah, N.A., Saleh, G. & Abdullah, T. (2013). Control of contamination and explant browning in Curculigo latifolia in vitro cultures. Journal of Medicinal Plants, 7, 448-454. https://doi.org/10.5897/JMPR012.859.
Basto S., Serrano C. & de Jaramillo E.H. (2012). Effects of donor plant age and explants on in vitro culture of Cedrela montana Moritz ex Turcz. Universitas Scientiarum, 17 (3), 263-271. http://www.scielo.org.co/scielo.php?script=sci_arttext&pid=S0122-74832012000300002&lng=en&nrm=iso
Beckman, C. H. (2000). Phenolic-storing cells: keys to programmed cell death and periderm formation in wilt disease resistance and in general defence responses in plants? Physiological and Molecular Plant Pathology, 57(3), 101–110. https://doi.org/10.1006/ pmpp.2000.0287
Benisheikh A.A.G., Tijani Y., Askira N.K., Waziri M.S. & Awana A.U. (2015). Prevalence and management of laboratory microbial contaminants in plant tissue culture. International Journal of Research, 2(6), 625-630.
Bhat, S.N.; Khalil, A.; Nazir, N.; Mir, M.A.; Khan, I.; Mubashir, S.S.; Dar, M.S.; Wani, S.H.; Hossain, M.A. (2022) In Vitro Prevention of Browning in Persian Walnut (Juglans regia L.) cv. Sulaiman. Int. J. Plant Biol., 13, 330-342. https://doi.org/10.3390/ijpb13030027
Cai, X., Wei, H., Liu, C., Ren, X., Thi, L. T., & Jeong, B. R. (2020). Synergistic Effect of NaCl Pretreatment and PVP on browning suppression and callus induction from petal explants of Paeonia lactiflora Pall. ‘Festival Maxima’. Plants (Basel, Switzerland), 9(3), 346. https://doi. org/10.3390/plants9030346
Chai, J., Gao, Y., Dong, Y., Kong, L., & Zhang, Y. (2018). Browning Treatment in Tissue Culture of ‘Hongyang’ Kiwifruit. IOP Conference Series Materials Science and Engineering, 452(2): 022075. https://doi.org/10.1088/1757-899X/452/2/022075
Devi S. & Yadav N.R. (2022) Role of explants collecting seasons and sterilization process on in vitro regeneration of guava: A technology hurdle. The Pharma Innovation Journal, 11(5): 1244-1248.
Dewan, A.; Nanda, K. & Gupta, S.C. (1992). In vitro micropropagation of Acacia nilotica subsp.indica Brenan via cotyledonary nodes. Pl. Cell Rep. 12: 18-21 https://doi.org/10.1007/BF00232415.
Dobránszki, J., & Teixeira da Silva, J. A. (2010). Micropropagation of apple—A review. Biotechnology Advances, 28(4), 462–488. https://doi.org/10.1016/j.biotechadv.20 10.02.008
Gioushy, S.F.E.; Liu, R.; Fan, H.K. A complete protocol to reduce browning during coconut (cocos nucifera L.) tissue culture through shoot tips and inflorescence explants. (2020) Plant Arch. 20, 2196–2204.
Huang L., Wang H., Shahid M.Q. & Zeng B. (2019). Chlorothalonil: an effective bacteriostatic agent for bud induction of Acacia auriculiformis under open condition (non-axenic). Plant Methods. 15:5. https://doi.org/10.1186/s13007-019-0390-3.
Irshad, M., Rizwan, H. M., Debnath, B., Anwar, M., Li, M., Liu, S., He, B., & Qiu, D. (2018). Ascorbic acid controls lethal browning and pluronic f-68 promotes high-frequency multiple shoot regeneration from cotyldonary node explant of okra (Abelmoschus esculentus L.). Hortscience, 53(1), 183–190. https://doi.org/10.21273/HORTSCI12315-18
Jones, A. M., & Saxena, P. K. (2013). Inhibition of phenylpropanoid biosynthesis in Artemisia annua L.: a novel approach to reduce oxidative browning in plant tissue culture. PloS one, 8(10): 76802. https://doi.org/10.1371/journal.pone.0076802
Krishna, H., Sairam, R. K., Singh, S. K., Patel, V. B., Sharma, R. R., Grover, M., Nain, L., & Sachdev, A. (2008). Mango explants browning: Effect of ontogenic age, mycorrhization and pre-treatments. Scientia Horticulturae, 118(2), 132-138. https://doi.org/10.1016/j.scienta.2008.05.040
Kuppusamy, S. Seenivasan R., Subramanian S., Chinnaraj, S., Rajasree S., Kathirvel, B., Thamaraiselvi, K. (2019) Optimizing the sterilization methods for initiation of the five different clones of the Eucalyptus hybrid species, Biocatalysis and Agricultural Biotechnology, Volume 22,101361, ISSN 1878-8181, https://doi.org/10.1016/j.bcab.2019.101361.
Martini, A. N., Papafotiou, M. & Vemmos, S. N. (2013). Season and Explant Origin Affect Phenolic Content, Browning of Explants, and Micropropagation of ×Malosorbus florentina (Zucc.) Browicz, HortScience horts, 48(1), 102-107. https://doi.org/10.21273/HORTSCI.48.1.102
Mohammadi, M., Bagheri, A., Marashi, H., Moshtaghi, N., & Balandari, A. (2011). Investigation into Seasonal Effect and Browning Inhibitor on Callus Regeneration of Seedless Barberry (Berberis vulgaris var. asperma). Plant Tissue Culture and Biotechnology, 21(2), 161–168. https://doi.org/10.3329/ptcb.v21i2.10239
Mudoi K.D., Saikia S.P. & Borthakur, M. (2014). Effect of nodal positions, seasonal variations, shoot clump and growth regulators on micropropagation of commercially important bamboo, Bambusa nutans Wall. ex. Munro. African Journal of Biotechnology, 13, 1961-1972. https://doi.org/10.5897/AJB2014.13659
Narang A. (2022). In vitro micropropagation of Acacia holosericea ex. G.Don through cotyledons. International Journal of Multidisciplinary Educational Research, 11(1,7), 70-75. http://ijmer.in.doi./2022/11.01.135
Ndakidemi, C. F., Mneney, E., & Ndakidemi, P. A. (2014). Effects of ascorbic acid in controlling lethal browning in in vitro culture of Brahylaena huillensis using nodal segments. American Journal of Plant Sciences 5(1), 187–191. https://doi.org/10.4236/ajps.2014.51024
Omamor I, Asemota, a, Eke, C, Eziashi, E. (2007). Fungal contaminants of the oil palm tissue culture in Nigerian institute for oil palm research (NIFOR) African Journal of Agricultural Research 2(10), 534-537.
Permadi, N., Nurzaman,M., Alhasnawi, A.N., Doni, F. & Julaeha, E.(2023). Managing Lethal Browning and Microbial Contamination in Musa spp. Tissue Culture: Synthesis and Perspectives. Horticulturae, 9(4), 453. https://doi.org/10.3390/horticulturae9040453
Sarkar P.K., Sinha A., Bhatt B.P., Kumar R., Sarkar A., Banerjee S., Ghosh J., Kujur R.S. & Kulkarni N. (2019) Effect of pretreatment of explants and in vitro culture media in controlling infection and browning in Kusum [Schleichera oleosa (Lour.) Oken] IJCS, SP6, 914-917. http://krishi.icar.gov.in/jspui/handle/123456789/53533
Selvarajan, E., Veena, R., & Kumar, N. M. (2008). Polyphenol oxidase, beyond enzyme browning, a review. Microbial Bioprospecting for Sustainable Development (pp. 203-222), Singapore, Springer
Shimelis, D., Bantte, K., & Feyissa, T. (2015). Effects of Polyvinyl Pyrrolidone and Activated Charcoal to Control Effect of Phenolic Oxidation on In Vitro Culture Establishment Stage of Micropropagation of Sugarcane (Saccharum Officinarum L.). Advances in Crop Science and Technology, 3(4), 184. https://doi.org/10.4172/2329-8863.1000184
Shukla, A., Narang A. & Kaur S. (2023). Effect of Adenine Sulphate on In Vitro Micropropagation of Acacia holosericea A. Cunn. ex G. Don Leaflets and Leaf Rachis Explants. Journal of Tropical Life Science, 13(3), 497–502. https://doi.org/10.11594/jtls.13.03.08.
Silva G.M., Xavier M.A., da Silva P.A., Saldanha C.W. & Stein V.C. (2020). Microbial contamination in micropropagation: causes and prevention. Plant Cell, Tissue and Organ Culture, 142(2), 215-227. https://doi.org/10.1007/s11240-020-01819-6.
Singh V., Ankur T., Chauhan P.K., Kumari, P. & Kaushal, S. (2011). Identification and prevention of bacterial contamination on explants used in plant tissue culture labs. International Journal of Pharmacy and Pharmaceutical Sciences, 3(4), 160-163.
Singh, N. V., Singh, S. K. & Patel, V. B. (2011). In vitro culture establishment studies on pomegranate. Indian Journal of Horticulture, 68(3), 307-311.
Singh, P., & Patel, R. M. (2016). Factors affecting in vitro degree of browning and culture establishment of pomegranate. African Journal of Plant Science, 10(2), 43-49. https://doi.org/10.5897/AJPS2013.1119
Sivanesan, I., Muthu, M., Gopal, J., Tasneem, S., Kim, D.H., Oh & J. W. (2021) A Fumigation-Based Surface Sterilization Approach for Plant Tissue Culture. Int J Environ Res Public Health, 18(5), 2282. https://doi.org/10.3390/ijerph18052282. PMID: 33668989; PMCID: PMC7967719.
Sultana J, Rahman M.M., Karim R., Khan R.H., & Hakim M.A. (2021). Seasonal variation in phytochemical constituents and antioxidant activity of Musa acuminata cv. Barjahaji: an approach towards micropropagation. Bangladesh Journal of Botany, 50(2), 291-300. https://doi.org/10.3329/bjb.v50i2.54100.
Tao, Fj., Zhang, Zy., Zhou, J. & Yao, N. & Wang, Dm. (2007). Contamination and browning in tissue culture of Platanus occidentalis L. Forestry Studies in China, 9: 279-282. https://doi.org/10.1007/s11632-007-0044-9.
Titov, S., Bhowmik, S. K., Mandal, A., Alam, M. S., & Uddin S. N. (2006). Control of phenolic compound secretion and effect of growth regulators for organ formation from Musa spp. cv. Kanthali floral bud explants. American Journal of Biochemistry and Biotechnology, 2(3), 97–104. https://doi.org/10.3844/ajbbsp.2006.97
TheodosyMsgoya, Helen Kanyaghu, Josiah Mutigitu, MatesoKulebelwa (2012). Identification and management of microbial contaminants of banana in vitro cultures, Tanzania, 55:3987-3994
Varghese, N. & Joy, P.P. (2016). Plant tissue culture contaminants identification and its response to fumigation. https://www.researchgate.net/publication/3 06034945
Yahia H A Elbasheer., Alem A A., Osman E I E (2019). Feasible Surface Sterilization Protocol for Microbial Contamination of Some Forest Trees on in vitro Cultures Explants. Acta Scientific Microbiology 2.5: 55-60.
Wang N., Yang C., Pan Z., Liu Y. & Peng S. (2015). Boron deficiency in woody plants: various responses and tolerance mechanisms. Front Plant Sci. 2, 916. https://doi.org/10.3389/fpls.2015.00916.
Zafar S., Abbasi B.H., & Aqil M. (2017). Influence of seasonal variation on the production of bioactive compounds and antioxidant activity in cell cultures of Cichorium intybus L. and Cynara cardunculus L. Journal of Applied Research on Medicinal and Aromatic Plants, 4, 9-18. https://doi.org/10.1016/j.jarmap.2016.09.002.
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How to Cite
Seasonal variation in contamination and browning of Acacia nilotica nodal explants in vitro. (2024). Journal of Applied and Natural Science, 16(1), 102-109. https://doi.org/10.31018/jans.v16i1.5303
