<i>In Vitro</i> Regeneration via Somatic Embryogenesis of <i>Schizozygia Coffeoides Baill</i> (Mpelepele)

Bashiloni Naaty; Cecilia Mweu Mbithe; Aggery Bernard Nyende; Peter Njenga; Joshua Kiilu Muli

doi:doi:10.11648/j.ajpb.20170202.14

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In Vitro Regeneration via Somatic Embryogenesis of Schizozygia Coffeoides Baill (Mpelepele)

Bashiloni Naaty, Cecilia Mweu Mbithe, Aggery Bernard Nyende, Peter Njenga, Joshua Kiilu Muli

Published in American Journal of Plant Biology (Volume 2, Issue 2)

Received: 20 March 2017 Accepted: 29 March 2017 Published: 19 April 2017

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Abstract

Schizozygia coffeoides (Mpelepele) is an endangered medicinal shrub with substantial antifungal and antibacterial properties. The sustainable utilization of this shrub is hindered by its limited regeneration potential since there exists no protocol for In vitro regeneration of not only this plant but the entire Apocynaceae family. The aim of this study was to develop an appropriate sterilization and regeneration protocol for In vitro regeneration of S. coffeoides using leaf discs, with the specific aims of determining the best NaOCL concentrations for explant sterilization as well as the appropriate plant growth hormone combinations for development of embryogenic calli, shoot development and rooting. Sterilization was optimized using NaOCl at the concentrations of 5.2mM, 7.8mM, 10.4mM and 13mM at exposure times of 10, 15, 20 and 25 minutes. The different concentrations of NaOCl had a significant effect on the survival of explants to sterilization (P=<0.0001), with the lower concentrations (5.2mM and 7.8mM) showing high levels of explant contamination and high concentration (13mM) leading to explant scotching. Similarly, the time explants were subjected to the sterilization substances was also found to significantly affect their ability to survive the sterilization process (P=0.0027). The best time and NaOCl concentration interaction for the sterilization of leaf explants was found out to be 10.4mM for 20 minutes. This time-NaOCl concentration interaction was found out to significantly produce a higher number of clean surviving explants than all other interactions (P=<0.0001). The best callus formation frequency of 68% was observed in MS media supplemented with 2.0mg/lBAP+0.8mg/lKin+0.4mg/l NAA+0.5mg/lTDZ, compared to a 39% callus formation frequency observed in media supplemented with 1.5mg/lBAP+0.8mg/lKin+0.4mg/lNAA+0.5mg/lTDZ and a 13% callus formation frequency observed in media supplemented with 1mg/lBAP+0.4mg/lKin+0.2mg/lNAA+0.05mg/lTDZ. Although somatic embryos formed in all media types, only media supplemented with 2.0mg/lBAP+0.8mg/lKin+0.4mg/lNAA+0.5mg/lTDZ formed most somatic embryos which survived to maturity and formed shoots, as most calli and embryos forming in other PGR supplementations died with subsequent subcultures. Roots only formed in shoots cultured in media supplemented with 1.0 BAP+0.5 IBA. The successful development of this regeneration protocol is expected to greatly contribute to mass production and conservation of this important shrub.

Published in	American Journal of Plant Biology (Volume 2, Issue 2)
DOI	10.11648/j.ajpb.20170202.14
Page(s)	66-72
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

Sterilization, Plant Growth Regulators, Callus Induction, Somatic Embryo

References

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[2]	Atanasov AG, Waltenberger B, Pferschy-Wenzig EM, Linder T, Wawrosch C, Uhrin P, Temml V, Wang L, Schwaiger S, Heiss EH, Rollinger JM, Schuster D, Breuss JM, Bochkov V, Mihovilovic MD, Kopp B, Bauer R, Dirsch VM, and Stuppner H. 2015. Discovery and resupply of pharmacologically active plant-derived natural products: A review. Biotechnol Adv 33: 1582–1614.
[3]	Magadula JJ, and Erasto P. 2009. Bioactive natural products derived from the East African flora. R Soc Chemestry 26: 1535–1554.
[4]	Kariba RM, Houghton PJ, and Yenesew A. 2002. Antimicrobial activities of a new schizozygane indoline alkaloid from Schizozygia coffaeoides and the revised structure of isoschizogaline. J Nat Prod 65: 566–569.
[5]	Maurice D, and Henri J. 1971. Contribution à l’inventaire des plantes médicinales de la côte Est de Madagascar. ORSTOm 8: 35–47.
[6]	Murashige T, and Skoog F. 1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant.
[7]	Jiménez VM. 2001. Regulation of in vitro somatic embryogenesis with emphasis on to the role of endogenous hormones. Rev Bras Fisiol Veg 13: 196–223.
[8]	Singh P, Singh A, Shukla AK, Singh L, Pande V, and Nailwal TK. 2009. Somatic embryogenesis and in vitro regeneration of an endangered medicinal plant sarpgandha (rauvolfia serpentina l.). Life Sci J 6: 57–62.
[9]	Ndakidemi CF, Mneney E, and Ndakidemi PA. 2013. Development of Sanitation Protocol for Leaf Explants of B. huillensis for in Vitro Culture. Am J Plant Sci 4: 2425–2430.
[10]	Çölgeçen H, Koca U, and Toker G. 2011. Influence of different sterilization methods on callus initiation and production of pigmented callus in Arnebia densiflora Ledeb. Tubitak 35: 513–520.
[11]	Tiwari V, Singh BD, and Tiwari KN. 1998. Shoot regeneration and somatic embryogenesis from different explants of Brahmi [Bacopa monniera (L.) Wettst.]. Plant Cell Rep 17: 538–543.
[12]	Wang HC, Chen JT, and Chang WC. 2006. Somatic embryogenesis and plant regeneration from leaf, root and stem-derived callus cultures of Areca catechu. Biol Plant 50: 279–282.
[13]	Othmani A, Bayoudh C, Drira N, Marrakchi M, and Trifi M. 2009. somatIC embryoGenesIs and plant reGeneratIon In Gymnema sylvestre. Plant Cell Tissue Organ Cult 38: 71–79.
[14]	Hatanaka T, Arakawa O, Yasuda T, Uchida N, and Yamaguchi T. 1991. Effect of plant growth regulators on somatic embryogenesis in leaf cultures of Coffea canephora. Plant Cell Rep 10: 179–182.

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Bashiloni Naaty, Cecilia Mweu Mbithe, Aggery Bernard Nyende, Peter Njenga, Joshua Kiilu Muli. (2017). In Vitro Regeneration via Somatic Embryogenesis of Schizozygia Coffeoides Baill (Mpelepele). American Journal of Plant Biology, 2(2), 66-72. https://doi.org/10.11648/j.ajpb.20170202.14

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ACS Style

Bashiloni Naaty; Cecilia Mweu Mbithe; Aggery Bernard Nyende; Peter Njenga; Joshua Kiilu Muli. In Vitro Regeneration via Somatic Embryogenesis of Schizozygia Coffeoides Baill (Mpelepele). Am. J. Plant Biol. 2017, 2(2), 66-72. doi: 10.11648/j.ajpb.20170202.14

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AMA Style

Bashiloni Naaty, Cecilia Mweu Mbithe, Aggery Bernard Nyende, Peter Njenga, Joshua Kiilu Muli. In Vitro Regeneration via Somatic Embryogenesis of Schizozygia Coffeoides Baill (Mpelepele). Am J Plant Biol. 2017;2(2):66-72. doi: 10.11648/j.ajpb.20170202.14

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@article{10.11648/j.ajpb.20170202.14,
  author = {Bashiloni Naaty and Cecilia Mweu Mbithe and Aggery Bernard Nyende and Peter Njenga and Joshua Kiilu Muli},
  title = {In Vitro Regeneration via Somatic Embryogenesis of Schizozygia Coffeoides Baill (Mpelepele)},
  journal = {American Journal of Plant Biology},
  volume = {2},
  number = {2},
  pages = {66-72},
  doi = {10.11648/j.ajpb.20170202.14},
  url = {https://doi.org/10.11648/j.ajpb.20170202.14},
  eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.ajpb.20170202.14},
  abstract = {Schizozygia coffeoides (Mpelepele) is an endangered medicinal shrub with substantial antifungal and antibacterial properties. The sustainable utilization of this shrub is hindered by its limited regeneration potential since there exists no protocol for In vitro regeneration of not only this plant but the entire Apocynaceae family. The aim of this study was to develop an appropriate sterilization and regeneration protocol for In vitro regeneration of S. coffeoides using leaf discs, with the specific aims of determining the best NaOCL concentrations for explant sterilization as well as the appropriate plant growth hormone combinations for development of embryogenic calli, shoot development and rooting. Sterilization was optimized using NaOCl at the concentrations of 5.2mM, 7.8mM, 10.4mM and 13mM at exposure times of 10, 15, 20 and 25 minutes. The different concentrations of NaOCl had a significant effect on the survival of explants to sterilization (P=<0.0001), with the lower concentrations (5.2mM and 7.8mM) showing high levels of explant contamination and high concentration (13mM) leading to explant scotching. Similarly, the time explants were subjected to the sterilization substances was also found to significantly affect their ability to survive the sterilization process (P=0.0027). The best time and NaOCl concentration interaction for the sterilization of leaf explants was found out to be 10.4mM for 20 minutes. This time-NaOCl concentration interaction was found out to significantly produce a higher number of clean surviving explants than all other interactions (P=<0.0001). The best callus formation frequency of 68% was observed in MS media supplemented with 2.0mg/lBAP+0.8mg/lKin+0.4mg/l NAA+0.5mg/lTDZ, compared to a 39% callus formation frequency observed in media supplemented with 1.5mg/lBAP+0.8mg/lKin+0.4mg/lNAA+0.5mg/lTDZ and a 13% callus formation frequency observed in media supplemented with 1mg/lBAP+0.4mg/lKin+0.2mg/lNAA+0.05mg/lTDZ. Although somatic embryos formed in all media types, only media supplemented with 2.0mg/lBAP+0.8mg/lKin+0.4mg/lNAA+0.5mg/lTDZ formed most somatic embryos which survived to maturity and formed shoots, as most calli and embryos forming in other PGR supplementations died with subsequent subcultures. Roots only formed in shoots cultured in media supplemented with 1.0 BAP+0.5 IBA. The successful development of this regeneration protocol is expected to greatly contribute to mass production and conservation of this important shrub.},
 year = {2017}
}

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TY  - JOUR
T1  - In Vitro Regeneration via Somatic Embryogenesis of Schizozygia Coffeoides Baill (Mpelepele)
AU  - Bashiloni Naaty
AU  - Cecilia Mweu Mbithe
AU  - Aggery Bernard Nyende
AU  - Peter Njenga
AU  - Joshua Kiilu Muli
Y1  - 2017/04/19
PY  - 2017
N1  - https://doi.org/10.11648/j.ajpb.20170202.14
DO  - 10.11648/j.ajpb.20170202.14
T2  - American Journal of Plant Biology
JF  - American Journal of Plant Biology
JO  - American Journal of Plant Biology
SP  - 66
EP  - 72
PB  - Science Publishing Group
SN  - 2578-8337
UR  - https://doi.org/10.11648/j.ajpb.20170202.14
AB  - Schizozygia coffeoides (Mpelepele) is an endangered medicinal shrub with substantial antifungal and antibacterial properties. The sustainable utilization of this shrub is hindered by its limited regeneration potential since there exists no protocol for In vitro regeneration of not only this plant but the entire Apocynaceae family. The aim of this study was to develop an appropriate sterilization and regeneration protocol for In vitro regeneration of S. coffeoides using leaf discs, with the specific aims of determining the best NaOCL concentrations for explant sterilization as well as the appropriate plant growth hormone combinations for development of embryogenic calli, shoot development and rooting. Sterilization was optimized using NaOCl at the concentrations of 5.2mM, 7.8mM, 10.4mM and 13mM at exposure times of 10, 15, 20 and 25 minutes. The different concentrations of NaOCl had a significant effect on the survival of explants to sterilization (P=<0.0001), with the lower concentrations (5.2mM and 7.8mM) showing high levels of explant contamination and high concentration (13mM) leading to explant scotching. Similarly, the time explants were subjected to the sterilization substances was also found to significantly affect their ability to survive the sterilization process (P=0.0027). The best time and NaOCl concentration interaction for the sterilization of leaf explants was found out to be 10.4mM for 20 minutes. This time-NaOCl concentration interaction was found out to significantly produce a higher number of clean surviving explants than all other interactions (P=<0.0001). The best callus formation frequency of 68% was observed in MS media supplemented with 2.0mg/lBAP+0.8mg/lKin+0.4mg/l NAA+0.5mg/lTDZ, compared to a 39% callus formation frequency observed in media supplemented with 1.5mg/lBAP+0.8mg/lKin+0.4mg/lNAA+0.5mg/lTDZ and a 13% callus formation frequency observed in media supplemented with 1mg/lBAP+0.4mg/lKin+0.2mg/lNAA+0.05mg/lTDZ. Although somatic embryos formed in all media types, only media supplemented with 2.0mg/lBAP+0.8mg/lKin+0.4mg/lNAA+0.5mg/lTDZ formed most somatic embryos which survived to maturity and formed shoots, as most calli and embryos forming in other PGR supplementations died with subsequent subcultures. Roots only formed in shoots cultured in media supplemented with 1.0 BAP+0.5 IBA. The successful development of this regeneration protocol is expected to greatly contribute to mass production and conservation of this important shrub.
VL  - 2
IS  - 2
ER  -

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Author Information

Bashiloni Naaty

Institute for Biotechnology Research, Jomo Kenyatta University of Agriculture and Technology, Nairobi, Kenya
Cecilia Mweu Mbithe

Institute for Biotechnology Research, Jomo Kenyatta University of Agriculture and Technology, Nairobi, Kenya
Aggery Bernard Nyende

Institute for Biotechnology Research, Jomo Kenyatta University of Agriculture and Technology, Nairobi, Kenya
Peter Njenga

Department of Botany, Jomo Kenyatta University of Agriculture and Technology, Nairobi, Kenya
Joshua Kiilu Muli

Department of Botany, Jomo Kenyatta University of Agriculture and Technology, Nairobi, Kenya

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APA Style

Bashiloni Naaty, Cecilia Mweu Mbithe, Aggery Bernard Nyende, Peter Njenga, Joshua Kiilu Muli. (2017). In Vitro Regeneration via Somatic Embryogenesis of Schizozygia Coffeoides Baill (Mpelepele). American Journal of Plant Biology, 2(2), 66-72. https://doi.org/10.11648/j.ajpb.20170202.14

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ACS Style

Bashiloni Naaty; Cecilia Mweu Mbithe; Aggery Bernard Nyende; Peter Njenga; Joshua Kiilu Muli. In Vitro Regeneration via Somatic Embryogenesis of Schizozygia Coffeoides Baill (Mpelepele). Am. J. Plant Biol. 2017, 2(2), 66-72. doi: 10.11648/j.ajpb.20170202.14

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AMA Style

Bashiloni Naaty, Cecilia Mweu Mbithe, Aggery Bernard Nyende, Peter Njenga, Joshua Kiilu Muli. In Vitro Regeneration via Somatic Embryogenesis of Schizozygia Coffeoides Baill (Mpelepele). Am J Plant Biol. 2017;2(2):66-72. doi: 10.11648/j.ajpb.20170202.14

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@article{10.11648/j.ajpb.20170202.14,
  author = {Bashiloni Naaty and Cecilia Mweu Mbithe and Aggery Bernard Nyende and Peter Njenga and Joshua Kiilu Muli},
  title = {In Vitro Regeneration via Somatic Embryogenesis of Schizozygia Coffeoides Baill (Mpelepele)},
  journal = {American Journal of Plant Biology},
  volume = {2},
  number = {2},
  pages = {66-72},
  doi = {10.11648/j.ajpb.20170202.14},
  url = {https://doi.org/10.11648/j.ajpb.20170202.14},
  eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.ajpb.20170202.14},
  abstract = {Schizozygia coffeoides (Mpelepele) is an endangered medicinal shrub with substantial antifungal and antibacterial properties. The sustainable utilization of this shrub is hindered by its limited regeneration potential since there exists no protocol for In vitro regeneration of not only this plant but the entire Apocynaceae family. The aim of this study was to develop an appropriate sterilization and regeneration protocol for In vitro regeneration of S. coffeoides using leaf discs, with the specific aims of determining the best NaOCL concentrations for explant sterilization as well as the appropriate plant growth hormone combinations for development of embryogenic calli, shoot development and rooting. Sterilization was optimized using NaOCl at the concentrations of 5.2mM, 7.8mM, 10.4mM and 13mM at exposure times of 10, 15, 20 and 25 minutes. The different concentrations of NaOCl had a significant effect on the survival of explants to sterilization (P=<0.0001), with the lower concentrations (5.2mM and 7.8mM) showing high levels of explant contamination and high concentration (13mM) leading to explant scotching. Similarly, the time explants were subjected to the sterilization substances was also found to significantly affect their ability to survive the sterilization process (P=0.0027). The best time and NaOCl concentration interaction for the sterilization of leaf explants was found out to be 10.4mM for 20 minutes. This time-NaOCl concentration interaction was found out to significantly produce a higher number of clean surviving explants than all other interactions (P=<0.0001). The best callus formation frequency of 68% was observed in MS media supplemented with 2.0mg/lBAP+0.8mg/lKin+0.4mg/l NAA+0.5mg/lTDZ, compared to a 39% callus formation frequency observed in media supplemented with 1.5mg/lBAP+0.8mg/lKin+0.4mg/lNAA+0.5mg/lTDZ and a 13% callus formation frequency observed in media supplemented with 1mg/lBAP+0.4mg/lKin+0.2mg/lNAA+0.05mg/lTDZ. Although somatic embryos formed in all media types, only media supplemented with 2.0mg/lBAP+0.8mg/lKin+0.4mg/lNAA+0.5mg/lTDZ formed most somatic embryos which survived to maturity and formed shoots, as most calli and embryos forming in other PGR supplementations died with subsequent subcultures. Roots only formed in shoots cultured in media supplemented with 1.0 BAP+0.5 IBA. The successful development of this regeneration protocol is expected to greatly contribute to mass production and conservation of this important shrub.},
 year = {2017}
}

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TY  - JOUR
T1  - In Vitro Regeneration via Somatic Embryogenesis of Schizozygia Coffeoides Baill (Mpelepele)
AU  - Bashiloni Naaty
AU  - Cecilia Mweu Mbithe
AU  - Aggery Bernard Nyende
AU  - Peter Njenga
AU  - Joshua Kiilu Muli
Y1  - 2017/04/19
PY  - 2017
N1  - https://doi.org/10.11648/j.ajpb.20170202.14
DO  - 10.11648/j.ajpb.20170202.14
T2  - American Journal of Plant Biology
JF  - American Journal of Plant Biology
JO  - American Journal of Plant Biology
SP  - 66
EP  - 72
PB  - Science Publishing Group
SN  - 2578-8337
UR  - https://doi.org/10.11648/j.ajpb.20170202.14
AB  - Schizozygia coffeoides (Mpelepele) is an endangered medicinal shrub with substantial antifungal and antibacterial properties. The sustainable utilization of this shrub is hindered by its limited regeneration potential since there exists no protocol for In vitro regeneration of not only this plant but the entire Apocynaceae family. The aim of this study was to develop an appropriate sterilization and regeneration protocol for In vitro regeneration of S. coffeoides using leaf discs, with the specific aims of determining the best NaOCL concentrations for explant sterilization as well as the appropriate plant growth hormone combinations for development of embryogenic calli, shoot development and rooting. Sterilization was optimized using NaOCl at the concentrations of 5.2mM, 7.8mM, 10.4mM and 13mM at exposure times of 10, 15, 20 and 25 minutes. The different concentrations of NaOCl had a significant effect on the survival of explants to sterilization (P=<0.0001), with the lower concentrations (5.2mM and 7.8mM) showing high levels of explant contamination and high concentration (13mM) leading to explant scotching. Similarly, the time explants were subjected to the sterilization substances was also found to significantly affect their ability to survive the sterilization process (P=0.0027). The best time and NaOCl concentration interaction for the sterilization of leaf explants was found out to be 10.4mM for 20 minutes. This time-NaOCl concentration interaction was found out to significantly produce a higher number of clean surviving explants than all other interactions (P=<0.0001). The best callus formation frequency of 68% was observed in MS media supplemented with 2.0mg/lBAP+0.8mg/lKin+0.4mg/l NAA+0.5mg/lTDZ, compared to a 39% callus formation frequency observed in media supplemented with 1.5mg/lBAP+0.8mg/lKin+0.4mg/lNAA+0.5mg/lTDZ and a 13% callus formation frequency observed in media supplemented with 1mg/lBAP+0.4mg/lKin+0.2mg/lNAA+0.05mg/lTDZ. Although somatic embryos formed in all media types, only media supplemented with 2.0mg/lBAP+0.8mg/lKin+0.4mg/lNAA+0.5mg/lTDZ formed most somatic embryos which survived to maturity and formed shoots, as most calli and embryos forming in other PGR supplementations died with subsequent subcultures. Roots only formed in shoots cultured in media supplemented with 1.0 BAP+0.5 IBA. The successful development of this regeneration protocol is expected to greatly contribute to mass production and conservation of this important shrub.
VL  - 2
IS  - 2
ER  -

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