Contact us
 |
[email protected] |
 |
3275638434 |
 |
 |
| Paper Publishing WeChat |
Useful Links
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License
Article
Author(s)
Heriberto García-Coronado1, Maria-Elena Báez-Flores2, Rosalba Troncoso-Rojas1, Marisela Rivera-Domínguez3 and Martín-Ernesto Tiznado-Hernández1
Full-Text PDF
XML 1168 Views
DOI:10.17265/2161-6264/2016.06.004
Affiliation(s)
1. Coordinación de Tecnología de Alimentos de Origen Vegetal, Centro de Investigación en Alimentación y Desarrollo A.C. (CIAD), Hermosillo, Sonora, CP 83304, México
2. Facultad de Ciencias Químico Biológicas, Universidad Autónoma de Sinaloa, Culiacán, Sinaloa, CP 80013, México
3. Coordinación de Ciencia de los Alimentos, Centro de Investigación en Alimentación y Desarrollo A.C. (CIAD), Hermosillo, Sonora, CP 83304, México
ABSTRACT
Mexico is a large producer of
table grape (Vitis vinifera L.) and therefore it is
important to develop protocols to store the grape varieties germplasm. The
objective of the present work was to design a protocol for the cryopreservation
by vitrification of zygotic embryos of V.
vinifera cv. “Red Globe” and evaluate
possible epigenetics changes. The plant vitrification solution 2 (PVS2) was
utilized before the utilization of liquid nitrogen (LN). The effect of this protocol on embryo
viability was tested by the triphenyl-tetrazolium chloride solution, as well as by the in vitro development of grape embryos
into plantlet. A cDNA expression library of grape zygotic embryos was created
to isolate expressed sequence tags of several DNA methyltrasferases. Gene expression of
domains rearranged methyltransferase type 1 (DMR1) and DNA (cytosine-5)-methyltransferase 1 (MET1-2) isozymes was analyzed by quantitative reverse transcriptase PCR.
The optimal conditions for vitrification were 10 min in 50% PVS2, followed by 10 min
in 100% PVS2. Under these conditions, about 30% of plantlet was obtained from
embryos after cryopreservation. It was recorded a reduction in the MET1-2 gene
expression, which plays a role
in the maintenance of DNA methylation. It is possible to cryopreserve viable
grape zygotic embryos, although the
treatment seems to induce
alterations in the normal DNA methylation pattern of the zygotic embryo genome.
KEYWORDS
Vitis vinifera, zygotic embryo, cryopreservation, vitrification, DNA methyltransferase.
Cite this paper
References