Comparison of two different micropropagation systems of Saccharum officinarum L. and expression analysis of PIP2;1 and EIN3 genes as efficiency system indicators
- 30 Downloads
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.
- Ali A, Naz S, Siddiqui FA, Iqbal J (2008) An efficient protocol for large scale production of sugarcane through micropropagation. Pakistan J Bot 40:139–149Google Scholar
- Baiges I, Schäffner AR, Affenzeller MJ, MA (2002) Plant aquaporins. Physiol Plant 115:175–182. https://doi.org/10.1034/j.1399-3054.2002.1150201.x CrossRefPubMedGoogle Scholar
- Bello-Bello JJ, Canto-Flick A, Balam-Uc E et al (2010) Improvement of in vitro proliferation and elongation of habanero pepper shoots (Capsicum chinense jacq.) by temporary immersion. HortScience 45:1093–1098Google Scholar
- Godoy-hernández G, Vázquez-flota FA (2006) Growth measurements. Plant Cell Cult Protoc 318:51–58Google Scholar
- Hake S, Smith HMS, Holtan H et al (2004) The role of knox genes in plant development. Annu Rev Cell Dev Biol 20:125–151. https://doi.org/10.1146/annurev.cellbio.20.031803.093824 CrossRefPubMedGoogle Scholar
- Houllou LM, De Souza RA (2015) Protocol optimization for in vitro sugarcane establishment from stem cuttings shoot tips. Res Biotechnol 6:42–48Google Scholar
- Peña-Ramírez YJ, Juárez-Gómez J, Gómez-López L et al (2010) Multiple adventitious shoot formation in Spanish Red Cedar (Cedrela odorata L.) cultured in vitro using juvenile and mature tissues: an improved micropropagation protocol for a highly valuable tropical tree species. In Vitro Cell Dev Biol 46:149–160. https://doi.org/10.1007/s11627-010-9280-0 CrossRefGoogle Scholar
- Pereira-Santana A, Alvarado-Robledo EJ, Zamora-Briseño JA, Ayala-Sumuano JT, González-Mendoza VM, Espadas-Gil F, Alcaráz LD, Castaño E, Keb-Llanes MA, Sanchez-Teyer F, Rodriguez-Zapata LC (2017) Transcriptional profiling of sugarcane leaves and roots under progressive osmotic stress reveals a regulated coordination of gene expression in a spatiotemporal manner. PlosOne 12(12):e0189271. https://doi.org/10.1371/journal.pone.0189271 CrossRefGoogle Scholar
- Rodríguez H, Ana M, Castillo C, Marco A, Flores BEP (2005) Genetic diversity of the most important sugar cane cultivars in Mexico. e-Gnosis 3:1–10Google Scholar
- Silva M (2016) In vitro propagation in temporary immersion system of sugarcane plants variety ‘RB 872552’ derived from somatic embryos. Biotecnología Vegetal 15:187–191Google Scholar
- United States Department of Agriculture (2017) Sugar: world markets and trade. United States Department of Agriculture, Washington, D.C.Google Scholar