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Effect of anti-ethylene compounds on isoenzyme patterns and genome stability during long term culture of Moringa oleifera

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Abstract

Multiplication of Moringa oleifera shoots on MS medium supplemented with 2.5 µM BAP for 3 weeks resulted in shoot vitrification which led to chlorosis, retardation of shoot formation, reduction in shoot length, necrosis of shoot tips and formation of friable calli on the base of cultured explants. Vitrification symptoms decreased when MS medium containing 2.5 µM BAP in combination with 10 µM AgNO3, 50 µM salicylic acid (SA) or 200 µM CoCl2 was used. Studying isoenzyme patterns of SOD, POX, CAT, GOT and EST indicated that moringa shoots multiplied without obvious variation in isoenzyme patterns up to 7 subcultures. Moringa shoots subjected to 14 subcultures and anti-ethylene compounds showed variation in isoenzyme patterns and were associated with the disappearance of vitrification which facilitated root formation and acclimatization. Under long term cultures, RAPD, ISSR and SSR indicated that AgNO3 was the optimal anti-ethylene substance for avoidance of vitrification in moringa but it resulted in high somaclonal variation. Application of SA decreased vitrification as well as somaclonal variation compared to CoCl2 under long term culture. Consequently, SA was recommended for moringa clonal multiplication.

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Abbreviations

BAP:

Benzyl amino purine

SA:

Salicylic acid

EST:

Esterases

GOT:

Glutamate oxaloactate transaminases

SOD:

Superoxide dismutases

CAT:

Catalases

POX:

Peroxidases

ISSR:

Simple Sequence Repeat

RAPD:

Random Amplified Polymorphic DNA

SSR:

Simple Sequence Repeat technique

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Acknowledgements

The authors would like to thank Prof. Moneim Shamloul of Fraunhofer USA Center for Molecular Biotechnology, Newark, DE, USA for English language revision.

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Authors and Affiliations

  1. Botany and Microbiology Department, Faculty of Science, Sohag University, Sohag, 82524, Egypt

    A. M. Hassanein, J. M. Salem, F. A. Faheed & A. El-nagish

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  1. A. M. Hassanein
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  2. J. M. Salem
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  3. F. A. Faheed
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  4. A. El-nagish
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Contributions

AMH proposed the idea and the experimental design, revised the manuscript and acted as a corresponding author; JMS participated in the experimental design, interpretation and acquisition the data; FAF participated in the experimental design and drafted the manuscript; AE performed the measurements, drafted the manuscript and performed the statistical analysis. All authors read and approved the final manuscript.

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Correspondence to A. M. Hassanein.

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The authors whose names are listed in this manuscript certify that they have NO affiliations with or involvement in any organization or entity with any financial interest in the subject matter or materials discussed in this manuscript.

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Communicated by Sergio J. Ochatt.

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Hassanein, A.M., Salem, J.M., Faheed, F.A. et al. Effect of anti-ethylene compounds on isoenzyme patterns and genome stability during long term culture of Moringa oleifera . Plant Cell Tiss Organ Cult 132, 201–212 (2018). https://doi.org/10.1007/s11240-017-1326-0

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