Osmo- and cryotherapy of sugarcane ( Saccharum spp. L.) shoot-tips infected with sugarcane mosaic virus (SCMV)
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ORIGINAL ARTICLE
Osmo- and cryotherapy of sugarcane (Saccharum spp. L.) shoot-tips infected with sugarcane mosaic virus (SCMV) Marı´a Teresa Gonza´lez-Arnao1 • Manuel Me´ndez-Cha´vez1 • Sugey Va´squez-Herna´ndez1 • Rosalı´a Nu´n˜ez-Pastrana2 • Carlos Alberto Cruz-Cruz1
Received: 9 April 2020 / Accepted: 30 September 2020 Indian Virological Society 2020
Abstract In this work, we investigated the effect of different osmoprotective treatments and of cryopreservation using a droplet-vitrification (D-V) protocol to eliminate sugarcane mosaic virus (SCMV) of shoot-tips excised from in vitro propagated infected plantlets. Shoot-tips of sugarcane (Saccharum spp. L.) were precultured on semisolid MS medium supplemented with 0.3 M sucrose for 1 day, loaded in solution with 0.4 M sucrose and 2 M glycerol for 30 min and exposed to plant vitrification solution 2 for 15 min at room temperature prior to ultra-rapid cooling in liquid nitrogen. Virus indexing was performed by the DASELISA immunoenzymatic test. The presence of SCMV was confirmed in the donor-plantlets derived of infected field material. No virus was detected in the regenerated plantlets from shoot-tips subjected to cryopreservation protocol. The progressive decrease in absorbances occurred from the first preculture treatment and no significant differences (P B 0.05) were found with respect to following steps of D-V protocol. These results indicate that the osmotic dehydration treatments (osmotherapy) and cryopreservation (cryotherapy) may be potentially effective strategies to remove the SCMV from infected plants. Keywords Sugarcane mosaic virus Virus eradication Droplet-vitrification Osmotherapy Cryotherapy
& Carlos Alberto Cruz-Cruz [email protected] 1
Facultad de Ciencias Quı´micas, Universidad Veracruzana, Prolongacio´n de Avenida Oriente 6, 1009, Rafael Alvarado, 94340 Orizaba, Veracruz, Mexico
2
Facultad de Ciencias Biolo´gicas y Agropecuarias, Universidad Veracruzana, Camino Pen˜uela-Amatla´n s/n, 94945 Municipio de Amatla´n de los Reyes, Veracruz, Mexico
Introduction Sugarcane (Saccharum spp. L.) is an increasingly important crop due to its agri-food and industrial importance for obtaining sugar and products such as alcohol, acetic acid, butanol, paper, compressed wood as well as enzymes and food for animals [31]. However, throughout its vegetative life cycle it is affected by a wide variety of pathogenic organisms, including fungi, bacteria and virus [23]. Among the viral diseases that produce the most economic affectations is the mosaic diseases, provoked by the sugarcane mosaic virus (SCMV). This disease is characterized by discolorations on the leafs blade reducing crop yield [29, 33, 47]. To date, there are no efficient chemical products to combat the virus in crops and the only thing that can be done to prevent them is the use of specific techniques, thus, it is very important to obtain material free of pathogens for increasing the production of agricultural crops, and to exchange vegetal germplasm [12, 13, 46]. Cryotherapy of shoot tips
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