Genetic Stability of Cassava Plants Regenerated Through Organogenesis Using Microsatellite Markers

Gilbert Osena; Nelson Onzere Amugune; Evans Nyaega Nyaboga

doi:doi:10.11648/j.jps.20170501.13

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Genetic Stability of Cassava Plants Regenerated Through Organogenesis Using Microsatellite Markers

Gilbert Osena, Nelson Onzere Amugune, Evans Nyaega Nyaboga

Published in Journal of Plant Sciences (Volume 5, Issue 1)

Received: 15 December 2016 Accepted: 30 December 2016 Published: 23 January 2017

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Abstract

Tissue culture technology of cassava (Manihot esculenta Crantz) is a viable alternative to currently adopted techniques for mass propagation, germplasm conservation and genetic improvement. However, somaclonal variation is a common phenomenon in tissue culture which makes it mandatory to monitor the genetic stability of plants. Therefore, the objective of this study was to evaluate the genetic stability of cassava plants regenerated from axillary bud explants through direct organogenesis using simple sequence repeat (SSR) markers. High shoot regeneration (81.2 – 90.0%) occurred in MS medium supplemented with 10 mg/L 6-bnzylaminopurine (BAP) and multiple shoots (2 – 4 shoots per from axillary bud explant) were formed for all the three cultivars (TME14, TMS60444 and Kibandameno) tested. High frequency of rooting (100%) was obtained after transferring the plantlets to cassava basic medium (CBM) and the rooted plants were successfully hardened and acclimatized in the glasshouse with 100% survival rate. Three-month old plants exhibited normal morphological characters comparing with the mother plant. A total of 10 SSR markers were used to validate the genetic homogeneity amongst five randomly selected plants along with the donor mother plants. DNA fingerprints of axillary bud regenerated plants displayed monomorphic bands similar to mother plant, indicating homogeneity among the regenerated plants with donor mother plant. The effect of subculture frequency on genetic stability of axillary bud-derived regenerants and micropropagated plants was also assessed using SSR markers. All the SSR profiles from axillary bud regenerants and micropropagated plants were monomorphic and comparable to mother plants from 1^st to 5^th subculture, confirming the genetic stability among clones and mother plants. At the 6^th subculture, similarity indicators between the progenies and the mother plants ranged from 0.95 to 1.0 and such a similarity indicated a very low polymorphism. The dendrograms generated through Unweighted Pair Group Method with arithmetic mean (UPGMA) analysis of the 6^th subculture revealed 96% similarity amongst axillary bud regenerants and micropropagated plants with donor mother plant. This low polymorphism ratio between mother plants, axillary bud regenerants and micro-propagated plants indicates the little effect of somaclonal variations, the high genetic similarity between mother plants and progenies and demonstrates the reliability of this propagation system for cassava. These results suggest that direct organogenesis from the axillary buds is the safest method for regeneration of true-to-type plants and this system can be used for clonal mass propagation, germplasm conservation and genetic transformation of cassava.

Published in	Journal of Plant Sciences (Volume 5, Issue 1)
DOI	10.11648/j.jps.20170501.13
Page(s)	19-28
Creative Commons	This is an Open Access article, distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution and reproduction in any medium or format, provided the original work is properly cited.
Copyright	Copyright © The Author(s), 2024. Published by Science Publishing Group

Keywords

Cassava, Axillary Buds, Genetic Stability, Simple Sequence Repeats, Micropropagation

References

[1]	Chetty, C., Rossin, C., Gruissem, W., Vanderschuren, H., and Rey, M. (2013) Empowering biotechnology in southern Africa: establishment of a robust transformation platform for the production of transgenic industry-preferred cassava. ‎N. Biotechnol. 30 (2): 136-143.
[2]	Burns, A., Roslyn, G., Julie, C., Anabela, Z., and Timothy, C. (2010) Cassava: The Drought, War and Famine Crop in a Changing World. Sustainability. 2 (11): 3572-3607.
[3]	Ceballos, H., Iglesias, C. A., Pérez, J. C., and Dixon, A. G. (2004) Cassava breeding: opportunities and challenges. Plant Mol Biol. 56 (4): 503-516.
[4]	Lebot, V. (2009) Tropical Root and Tuber Crops; Cassava, sweet potato, yams and aroids. Cabi, p. 434.
[5]	Jansson, C., Westerbergh, A., Zhang, J., Hu, X., and Sun, C. (2009) Cassava, a potential biofuel crop in (the) People’s Republic of China. Appl. Energy, 86: S95-S99.
[6]	FAO. (2013) The State of Food Insecurity in the World 2013. The multiple dimensions of food security. 2013, FAO: ROME.
[7]	Da Silva, R. M., Bandel, G., and Martins, P. S. (2003) Mating system in an experimental garden composed of cassava (Manihot esculenta Crantz) ethnovarieties. Euphytica, 34 (2): 127-135.
[8]	Puonti-Kaerlas, J. (1998) Cassava biotechnology. Biotechnol Genet Eng Rev. 15 (1): 329-364.
[9]	Chavarriaga-Aguirre, P., Brand, A., Medina, A., Prías, M., Escobar, R., Martinez, J., Díaz, P., López, C., Roca, W. M., and Tohme, J. (2016) The potential of using biotechnology to improve cassava: a review. In Vitro Cell Dev. Biol. Plant. 52 (5): 461-478.
[10]	Nkaa, F., Ene-Obong, E., Taylor, N., Fauquet, C., and Mbanaso, E. (2013) Elimination of African Cassava Mosaic Virus (ACMV) and East African Cassava Mosaic Virus (EACMV) from cassava (Manihot esculenta Crantz) cv.‘Nwugo’ via somatic embryogenesis. Am. J. Biotechnol. Mol. Sci. 3 (2): 33-40.
[11]	Mapayi, E., Ojo, D., Oduwaye, O., and Porbeni, J. (2013) Optimization of in vitro propagation of cassava (Manihot esculenta Crantz) Genotypes. J. Agric. Sci. 5 (3): 261.
[12]	Neelakandan, A. K., and Wang, K. (2012) Recent progress in the understanding of tissue culture-induced genome level changes in plants and potential applications. Plant Cell Rep. 31 (4): 597-620.
[13]	Martins, M., Sarmento, D., and Oliveira, M. (2004). Genetic stability of micropropagated almond plantlets, as assessed by RAPD and ISSR markers. Plant Cell Rep. 23 (7): 492-496.
[14]	Saker, M., Bekheet, S., Taha, H., Fahmy, A., and Moursy, H. (2000) Detection of somaclonal variations in tissue culture-derived date palm plants using isoenzyme analysis and RAPD fingerprints. Biol. Plantarum. 43 (3): 347-351.
[15]	Vidal, Á., Vieira, L., Ferreira, C., Souza, F., Souza, A., and Ledo, C. (2015) Genetic fidelity and variability of micropropagated cassava plants (Manihot esculenta Crantz) evaluated using ISSR markers. Genet Mol Biol. 14 (3): 7759-7770.
[16]	Varshney, R. K., Graner, A., and Sorrells, M. E. (2005) Genetic microsatellite markers in plants: features and applications. Trends Biotechnol. 23 (1): 48-55.
[17]	Nookaraju, A., and Agrawal, D. (2012) Genetic homogeneity of in vitro raised plants of grapevine cv. Crimson Seedless revealed by ISSR and microsatellite markers. S Afr J Bot. 78: 302-306.
[18]	Rahman, M., and Rajora, O. (2001) Microsatellite DNA somaclonal variation in micropropagated trembling aspen (Populus tremuloides). Plant Cell Rep. 20 (6): 531-536.
[19]	Marum, L., Rocheta, M., Maroco, J., Oliveira, M. M., and Miguel, C. (2009) Analysis of genetic stability at SSR loci during somatic embryogenesis in maritime pine (Pinus pinaster). Plant Cell Rep. 28 (4): 673-682.
[20]	Mtunguja, M. K., Laswai, H. S., Kanju, E., Ndunguru, J., and Muzanila, Y. C. (2016) Effect of genotype and genotype by environment interaction on total cyanide content, fresh root, and starch yield in farmer‐preferred cassava landraces in Tanzania. Food Sci. Nutr. 4(6): 791–801.
[21]	Alicai T, Omongo CA, Kawuki R, Pariyo A, Baguma Y, Bua A (2010) National Cassava Programme-Uganda, 2009 National Survey.
[22]	Zainuddin, I. M., Schlegel, K., Gruissem, W., and Vanderschuren, H. (2012) Robust transformation procedure for the production of transgenic farmer-preferred cassava landraces. Plant methods. 8: 24
[23]	Nyaboga, E., Njiru, J., and Tripathi, L. (2015) Factors influencing somatic embryogenesis, regeneration, and Agrobacterium-mediated transformation of cassava (Manihot esculenta Crantz) cultivar TME14. Front. Plant Sci. 6: 411.
[24]	Sharma, K., Mishra, A. K., and Misra, R. S. (2008) A simple and efficient method for extraction of genomic DNA from tropical tuber crops. Afr. J. Biotechnol. 7 (8): 1018-1022.
[25]	Garcia-Vallvé, S., Palau, J., and Romeu, A. (1999) Horizontal gene transfer in glycosyl hydrolases inferred from codon usage in Escherichia coli and Bacillus subtilis. Mol. Biol. Evol., 16 (9): 1125-1134.
[26]	Jaccard, P. (1908) Nouvelles recherches sur Ia distribution Rorale. Bull. Soc. vaud. Sci. nat. 44: 223-270.
[27]	Wasswa, P., Alicai, A., and Mukasa, S. (2010) Optimisation of in vitro techniques for Cassava brown streak virus elimination from infected cassava clones. Afr. Crop Sci. J., 18 (4): 235-241.
[28]	Debnath, S. C. (2005) A two-step procedure for adventitious shoot regeneration from in vitro-derived lingonberry leaves: shoot induction with TDZ and shoot elongation using zeatin. HortScience, 40(1): 189-192.
[29]	Vázquez, A. and R. Linacero. (2010) Stress and somaclonal variation, in Plant Developmental Biology-Biotechnological Perspectives. Kluwer, Dordrecht, Netherlands p. 45-64.
[30]	Zilberman, D., and Henikoff, S. (2007) Genome-wide analysis of DNA methylation patterns. Development. Development, 134(22): 3959-396.
[31]	Escobar, R., L. Munoz, and W. Roca. (2009) Cassava micropropagation for rapid seed production using temporary immersion bioreactors. International Center of Tropical Agriculture (CIAT). 2009: Cali, Columbia.
[32]	Konan, N. K., R. S. Sangwan, and B. S. Sangwan-Norreel. (2006) Efficient in iitro shoot regeneration systems in cassava (Manihot esculenta Crantz). Plant Breed. 113 (3): 227-236.
[33]	Medina, R. D., Faloci, M. M., Gonzalez, A. M., and Mroginski, L. A. (2006). In vitro cultured primary roots derived from stem segments of cassava (Manihot esculenta) can behave like storage organs. Ann Bot. 99(3): 409-423.
[34]	Ostry, M., Hackett, W., Michler, C., Serres, R., and McCown, B. (1994) Influence of regeneration method and tissue source on the frequency of somatic variation in Populus to infection by Septoria musiva. Plant Sci. 97 (2): 209-215.
[35]	Wang, P.-J., and Charles, A. (1991) Micropropagation through meristem culture High-Tech and Micropropagation I. Kluwer, Dordrecht, Netherlands p. 32-52.
[36]	Raji, A. A., Anderson, J. V., Kolade, O. A., Ugwu, C. D., Dixon, A. G., and Ingelbrecht, I. L. (2009) Gene-based microsatellites for cassava (Manihot esculenta Crantz): prevalence, polymorphisms, and cross-taxa utility. BMC Plant Biol. 9 (1): 1.
[37]	Bhatia, R., Singh, K., Sharma, T., and Jhang, T. (2011). Evaluation of the genetic fidelity of in vitro propagated gerbera (Gerbera jamesonii Bolus) using DNA-based markers. Plant Cell Tissue Organ Cult. 104 (1): 131-135.
[38]	Kumar, N., Modi, A. R., Singh, A. S., Gajera, B. B., Patel, A. R., Patel, M. P., and Subhash, N. (2010) Assessment of genetic fidelity of micropropagated date palm (Phoenix dactylifera L.) plants by RAPD and ISSR markers assay. Physiol Mol Biol Plants. 16(2): 207-213.
[39]	Rani, V., and Raina, S. (2000) Genetic fidelity of organized meristem-derived micropropagated plants: a critical reappraisal. In Vitro Cell Dev Biol Plant. 36 (5): 319-330.
[40]	Joshi, P., and Dhawan, V. (2007) Assessment of genetic fidelity of micropropagated Swertia chirayita plantlets by ISSR marker assay. Biol Plantarum, 51 (1): 22-26.
[41]	Cassells, A. C., and Curry, R. F. (2001) Oxidative stress and physiological, epigenetic and genetic variability in plant tissue culture: implications for micropropagators and genetic engineers. Plant Cell, Tissue and Organ Cult. 64 (2-3): 145-157.
[42]	Thiem, B., Kikowska, M., Krawczyk, A., Więckowska, B., and Sliwinska, E. (2013) Phenolic acid and DNA contents of micropropagated Eryngium planum L. Plant Cell, Tissue and Organ Cult. 114 (2): 197-206.
[43]	Devi, S. P., Kumaria, S., Rao, S. R., and Tandon, P. (2015). Genetic fidelity assessment in micropropagated plants using cytogenetical analysis and heterochromatin distribution: a case study with Nepenthes khasiana Hook f. Protoplasma. 252 (5): 1305-1312.
[44]	Us-Camas, R., Rivera-Solís, G., Duarte-Aké, F., and De-la-Pena, C. (2014) In vitro culture: an epigenetic challenge for plants. Plant Cell, Tissue and Organ Cult. 118 (2): 187-201.
[45]	Miguel, C., and Marum, L. (2011) An epigenetic view of plant cells cultured in vitro: somaclonal variation and beyond. J Exp Bot. 62 (11): 3713-3725.
[46]	Modgil, M., Mahajan, K., Chakrabarti, S., Sharma, D., and Sobti, R. (2005) Molecular analysis of genetic stability in micropropagated apple rootstock MM106. Sci Hort. 104 (2): 151-160.
[47]	Jackson, A. L., Chen, R., and Loeb, L. A. (1998) Induction of microsatellite instability by oxidative DNA damage. Proc. Natl. Acad. Sci. U. S. A. 95 (21): 12468-12473.

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Gilbert Osena, Nelson Onzere Amugune, Evans Nyaega Nyaboga. (2017). Genetic Stability of Cassava Plants Regenerated Through Organogenesis Using Microsatellite Markers. Journal of Plant Sciences, 5(1), 19-28. https://doi.org/10.11648/j.jps.20170501.13

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Gilbert Osena; Nelson Onzere Amugune; Evans Nyaega Nyaboga. Genetic Stability of Cassava Plants Regenerated Through Organogenesis Using Microsatellite Markers. J. Plant Sci. 2017, 5(1), 19-28. doi: 10.11648/j.jps.20170501.13

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AMA Style

Gilbert Osena, Nelson Onzere Amugune, Evans Nyaega Nyaboga. Genetic Stability of Cassava Plants Regenerated Through Organogenesis Using Microsatellite Markers. J Plant Sci. 2017;5(1):19-28. doi: 10.11648/j.jps.20170501.13

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@article{10.11648/j.jps.20170501.13,
  author = {Gilbert Osena and Nelson Onzere Amugune and Evans Nyaega Nyaboga},
  title = {Genetic Stability of Cassava Plants Regenerated Through Organogenesis Using Microsatellite Markers},
  journal = {Journal of Plant Sciences},
  volume = {5},
  number = {1},
  pages = {19-28},
  doi = {10.11648/j.jps.20170501.13},
  url = {https://doi.org/10.11648/j.jps.20170501.13},
  eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.jps.20170501.13},
  abstract = {Tissue culture technology of cassava (Manihot esculenta Crantz) is a viable alternative to currently adopted techniques for mass propagation, germplasm conservation and genetic improvement. However, somaclonal variation is a common phenomenon in tissue culture which makes it mandatory to monitor the genetic stability of plants. Therefore, the objective of this study was to evaluate the genetic stability of cassava plants regenerated from axillary bud explants through direct organogenesis using simple sequence repeat (SSR) markers. High shoot regeneration (81.2 – 90.0%) occurred in MS medium supplemented with 10 mg/L 6-bnzylaminopurine (BAP) and multiple shoots (2 – 4 shoots per from axillary bud explant) were formed for all the three cultivars (TME14, TMS60444 and Kibandameno) tested. High frequency of rooting (100%) was obtained after transferring the plantlets to cassava basic medium (CBM) and the rooted plants were successfully hardened and acclimatized in the glasshouse with 100% survival rate. Three-month old plants exhibited normal morphological characters comparing with the mother plant. A total of 10 SSR markers were used to validate the genetic homogeneity amongst five randomly selected plants along with the donor mother plants. DNA fingerprints of axillary bud regenerated plants displayed monomorphic bands similar to mother plant, indicating homogeneity among the regenerated plants with donor mother plant. The effect of subculture frequency on genetic stability of axillary bud-derived regenerants and micropropagated plants was also assessed using SSR markers. All the SSR profiles from axillary bud regenerants and micropropagated plants were monomorphic and comparable to mother plants from 1st  to 5th  subculture, confirming the genetic stability among clones and mother plants. At the 6th subculture, similarity indicators between the progenies and the mother plants ranged from 0.95 to 1.0 and such a similarity indicated a very low polymorphism. The dendrograms generated through Unweighted Pair Group Method with arithmetic mean (UPGMA) analysis of the 6th subculture revealed 96% similarity amongst axillary bud regenerants and micropropagated plants with donor mother plant. This low polymorphism ratio between mother plants, axillary bud regenerants and micro-propagated plants indicates the little effect of somaclonal variations, the high genetic similarity between mother plants and progenies and demonstrates the reliability of this propagation system for cassava. These results suggest that direct organogenesis from the axillary buds is the safest method for regeneration of true-to-type plants and this system can be used for clonal mass propagation, germplasm conservation and genetic transformation of cassava.},
 year = {2017}
}

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TY  - JOUR
T1  - Genetic Stability of Cassava Plants Regenerated Through Organogenesis Using Microsatellite Markers
AU  - Gilbert Osena
AU  - Nelson Onzere Amugune
AU  - Evans Nyaega Nyaboga
Y1  - 2017/01/23
PY  - 2017
N1  - https://doi.org/10.11648/j.jps.20170501.13
DO  - 10.11648/j.jps.20170501.13
T2  - Journal of Plant Sciences
JF  - Journal of Plant Sciences
JO  - Journal of Plant Sciences
SP  - 19
EP  - 28
PB  - Science Publishing Group
SN  - 2331-0731
UR  - https://doi.org/10.11648/j.jps.20170501.13
AB  - Tissue culture technology of cassava (Manihot esculenta Crantz) is a viable alternative to currently adopted techniques for mass propagation, germplasm conservation and genetic improvement. However, somaclonal variation is a common phenomenon in tissue culture which makes it mandatory to monitor the genetic stability of plants. Therefore, the objective of this study was to evaluate the genetic stability of cassava plants regenerated from axillary bud explants through direct organogenesis using simple sequence repeat (SSR) markers. High shoot regeneration (81.2 – 90.0%) occurred in MS medium supplemented with 10 mg/L 6-bnzylaminopurine (BAP) and multiple shoots (2 – 4 shoots per from axillary bud explant) were formed for all the three cultivars (TME14, TMS60444 and Kibandameno) tested. High frequency of rooting (100%) was obtained after transferring the plantlets to cassava basic medium (CBM) and the rooted plants were successfully hardened and acclimatized in the glasshouse with 100% survival rate. Three-month old plants exhibited normal morphological characters comparing with the mother plant. A total of 10 SSR markers were used to validate the genetic homogeneity amongst five randomly selected plants along with the donor mother plants. DNA fingerprints of axillary bud regenerated plants displayed monomorphic bands similar to mother plant, indicating homogeneity among the regenerated plants with donor mother plant. The effect of subculture frequency on genetic stability of axillary bud-derived regenerants and micropropagated plants was also assessed using SSR markers. All the SSR profiles from axillary bud regenerants and micropropagated plants were monomorphic and comparable to mother plants from 1st  to 5th  subculture, confirming the genetic stability among clones and mother plants. At the 6th subculture, similarity indicators between the progenies and the mother plants ranged from 0.95 to 1.0 and such a similarity indicated a very low polymorphism. The dendrograms generated through Unweighted Pair Group Method with arithmetic mean (UPGMA) analysis of the 6th subculture revealed 96% similarity amongst axillary bud regenerants and micropropagated plants with donor mother plant. This low polymorphism ratio between mother plants, axillary bud regenerants and micro-propagated plants indicates the little effect of somaclonal variations, the high genetic similarity between mother plants and progenies and demonstrates the reliability of this propagation system for cassava. These results suggest that direct organogenesis from the axillary buds is the safest method for regeneration of true-to-type plants and this system can be used for clonal mass propagation, germplasm conservation and genetic transformation of cassava.
VL  - 5
IS  - 1
ER  -

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Author Information

Gilbert Osena

School of Biological Sciences, University of Nairobi, Nairobi, Kenya
Nelson Onzere Amugune

School of Biological Sciences, University of Nairobi, Nairobi, Kenya
Evans Nyaega Nyaboga

Department of Biochemistry, University of Nairobi, Nairobi, Kenya

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APA Style

Gilbert Osena, Nelson Onzere Amugune, Evans Nyaega Nyaboga. (2017). Genetic Stability of Cassava Plants Regenerated Through Organogenesis Using Microsatellite Markers. Journal of Plant Sciences, 5(1), 19-28. https://doi.org/10.11648/j.jps.20170501.13

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ACS Style

Gilbert Osena; Nelson Onzere Amugune; Evans Nyaega Nyaboga. Genetic Stability of Cassava Plants Regenerated Through Organogenesis Using Microsatellite Markers. J. Plant Sci. 2017, 5(1), 19-28. doi: 10.11648/j.jps.20170501.13

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AMA Style

Gilbert Osena, Nelson Onzere Amugune, Evans Nyaega Nyaboga. Genetic Stability of Cassava Plants Regenerated Through Organogenesis Using Microsatellite Markers. J Plant Sci. 2017;5(1):19-28. doi: 10.11648/j.jps.20170501.13

Copy | Download

@article{10.11648/j.jps.20170501.13,
  author = {Gilbert Osena and Nelson Onzere Amugune and Evans Nyaega Nyaboga},
  title = {Genetic Stability of Cassava Plants Regenerated Through Organogenesis Using Microsatellite Markers},
  journal = {Journal of Plant Sciences},
  volume = {5},
  number = {1},
  pages = {19-28},
  doi = {10.11648/j.jps.20170501.13},
  url = {https://doi.org/10.11648/j.jps.20170501.13},
  eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.jps.20170501.13},
  abstract = {Tissue culture technology of cassava (Manihot esculenta Crantz) is a viable alternative to currently adopted techniques for mass propagation, germplasm conservation and genetic improvement. However, somaclonal variation is a common phenomenon in tissue culture which makes it mandatory to monitor the genetic stability of plants. Therefore, the objective of this study was to evaluate the genetic stability of cassava plants regenerated from axillary bud explants through direct organogenesis using simple sequence repeat (SSR) markers. High shoot regeneration (81.2 – 90.0%) occurred in MS medium supplemented with 10 mg/L 6-bnzylaminopurine (BAP) and multiple shoots (2 – 4 shoots per from axillary bud explant) were formed for all the three cultivars (TME14, TMS60444 and Kibandameno) tested. High frequency of rooting (100%) was obtained after transferring the plantlets to cassava basic medium (CBM) and the rooted plants were successfully hardened and acclimatized in the glasshouse with 100% survival rate. Three-month old plants exhibited normal morphological characters comparing with the mother plant. A total of 10 SSR markers were used to validate the genetic homogeneity amongst five randomly selected plants along with the donor mother plants. DNA fingerprints of axillary bud regenerated plants displayed monomorphic bands similar to mother plant, indicating homogeneity among the regenerated plants with donor mother plant. The effect of subculture frequency on genetic stability of axillary bud-derived regenerants and micropropagated plants was also assessed using SSR markers. All the SSR profiles from axillary bud regenerants and micropropagated plants were monomorphic and comparable to mother plants from 1st  to 5th  subculture, confirming the genetic stability among clones and mother plants. At the 6th subculture, similarity indicators between the progenies and the mother plants ranged from 0.95 to 1.0 and such a similarity indicated a very low polymorphism. The dendrograms generated through Unweighted Pair Group Method with arithmetic mean (UPGMA) analysis of the 6th subculture revealed 96% similarity amongst axillary bud regenerants and micropropagated plants with donor mother plant. This low polymorphism ratio between mother plants, axillary bud regenerants and micro-propagated plants indicates the little effect of somaclonal variations, the high genetic similarity between mother plants and progenies and demonstrates the reliability of this propagation system for cassava. These results suggest that direct organogenesis from the axillary buds is the safest method for regeneration of true-to-type plants and this system can be used for clonal mass propagation, germplasm conservation and genetic transformation of cassava.},
 year = {2017}
}

Copy | Download

TY  - JOUR
T1  - Genetic Stability of Cassava Plants Regenerated Through Organogenesis Using Microsatellite Markers
AU  - Gilbert Osena
AU  - Nelson Onzere Amugune
AU  - Evans Nyaega Nyaboga
Y1  - 2017/01/23
PY  - 2017
N1  - https://doi.org/10.11648/j.jps.20170501.13
DO  - 10.11648/j.jps.20170501.13
T2  - Journal of Plant Sciences
JF  - Journal of Plant Sciences
JO  - Journal of Plant Sciences
SP  - 19
EP  - 28
PB  - Science Publishing Group
SN  - 2331-0731
UR  - https://doi.org/10.11648/j.jps.20170501.13
AB  - Tissue culture technology of cassava (Manihot esculenta Crantz) is a viable alternative to currently adopted techniques for mass propagation, germplasm conservation and genetic improvement. However, somaclonal variation is a common phenomenon in tissue culture which makes it mandatory to monitor the genetic stability of plants. Therefore, the objective of this study was to evaluate the genetic stability of cassava plants regenerated from axillary bud explants through direct organogenesis using simple sequence repeat (SSR) markers. High shoot regeneration (81.2 – 90.0%) occurred in MS medium supplemented with 10 mg/L 6-bnzylaminopurine (BAP) and multiple shoots (2 – 4 shoots per from axillary bud explant) were formed for all the three cultivars (TME14, TMS60444 and Kibandameno) tested. High frequency of rooting (100%) was obtained after transferring the plantlets to cassava basic medium (CBM) and the rooted plants were successfully hardened and acclimatized in the glasshouse with 100% survival rate. Three-month old plants exhibited normal morphological characters comparing with the mother plant. A total of 10 SSR markers were used to validate the genetic homogeneity amongst five randomly selected plants along with the donor mother plants. DNA fingerprints of axillary bud regenerated plants displayed monomorphic bands similar to mother plant, indicating homogeneity among the regenerated plants with donor mother plant. The effect of subculture frequency on genetic stability of axillary bud-derived regenerants and micropropagated plants was also assessed using SSR markers. All the SSR profiles from axillary bud regenerants and micropropagated plants were monomorphic and comparable to mother plants from 1st  to 5th  subculture, confirming the genetic stability among clones and mother plants. At the 6th subculture, similarity indicators between the progenies and the mother plants ranged from 0.95 to 1.0 and such a similarity indicated a very low polymorphism. The dendrograms generated through Unweighted Pair Group Method with arithmetic mean (UPGMA) analysis of the 6th subculture revealed 96% similarity amongst axillary bud regenerants and micropropagated plants with donor mother plant. This low polymorphism ratio between mother plants, axillary bud regenerants and micro-propagated plants indicates the little effect of somaclonal variations, the high genetic similarity between mother plants and progenies and demonstrates the reliability of this propagation system for cassava. These results suggest that direct organogenesis from the axillary buds is the safest method for regeneration of true-to-type plants and this system can be used for clonal mass propagation, germplasm conservation and genetic transformation of cassava.
VL  - 5
IS  - 1
ER  -

Copy | Download