Comparison of two different micropropagation systems of Saccharum officinarum L. and expression analysis of PIP2;1 and EIN3 genes as efficiency system indicators
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In sugarcane commercial plantations, seedlings are obtained by vegetative propagation from stem segments. However, this form of seedling production has a downside as it is responsible for spreading various pathogens that accumulate in plants during the cultivation cycle. The development of a disease-free and pathogen-free large-scale production protocol is useful to help minimize the spread of such diseases in commercial plantations. For this reason, a plant tissue culture methodology such as in vitro micropropagation offers the best alternative. In vitro micropropagation protocols using bioreactors based on the temporary immersion system such as BioMINT II™ is a cost-effective design. Our aim was to evaluate the efficiency of two different systems of in vitro micropropagation for sugarcane seedlings; semi-solid and BioMINT II™ in terms of biomass production, morphological and physiological parameters. Beside this, we tested by RT-qPCR two genes (PIP2;1 and EIN3) involved in plant development, as possible molecular markers for quality analysis of development during the tested in vitro micropropagation conditions. At 28 days we obtained more shoots and better quality physiological parameters in the BioMINT II than in the semi-solid system. On the other hand, we characterized and correlated the expression of PIP2;1 and EIN3 with the morphological parameters of the two systems. Our results give a better option for biomass production with the BioMINT II™ and suggest additional experiments of the PIP2;1 and EIN3 genes to be considered as molecular markers for quality analysis of physiological parameters during in vitro micropropagation.
KeywordsSugarcane BioMINT II (Bioreactor Modular Temporary Immersion) Micropropagation system Magenta box 6-Benzylaminopurine (BAP) RT-qPCR
Murashige and Skoog
Temporary immersion system
- BioMINT II
Modular immersion bioreactor
Root apical meristem
Gross domestic product
Shoot apical meristem
We would like to thank “Colegio de Posgraduados (Colpos)” Campus Campeche, Mexico, for providing plant material. The authors would also like to thank the Centro de Investigación Científica de Yucatán, Mexico, for supporting this research. We thank the anonymous referees for their constructive comments and suggestions.
EACB, MAHA, and LCRZ, conceived and designed the research; APS and EACB performed sample preparation and total RNA extraction; APS performed bioinformatic analysis from the sugarcane transcriptome to obtain the candidate sequences for the RT-qPCR experiment. VMGM designed the primers of RT-qPCR. MAHA performed the morphological evaluations and the massive micropropagation of the plants; MAKL in vitro adaptation of sugarcane plants; EACB and VMGM performed the RT-qPCR experiment and analysed the data; MAHA and EACB wrote the document. LCRZ, MLR, APS, and EC participated in the critical review of important intellectual content and reviewed the final version of the manuscript. All authors read and approved the final manuscript.
This work was supported by the SEP-CONACYT (No.: 215098). EACB received the financial support of a scholarship (462859) from CONACYT to obtain her Master degree and perform the experiments during her studies.
Compliance with ethical standards
Conflict of interest
The authors declare that they have not competing interests.
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