Visiting Scientists

Fernanda Souza

Associated to Labex Scientist Alfredo Augusto Alves

Cryopreservation techniques for Pineapple and Citrus

Research team: Fernanda Souza – Embrapa Cassava & Fruits, Cruz das Almas BA; Alfredo Alves – Embrapa/LABEX, Fort Collins CO; Ergun Kaya – Gebze Institute of Technology-Kocaeli-Turkey and NCGRP/ARS, USA; David Ellis – NCGRP/ARS, Fort Collins CO; and Maria Jenderek – NCGRP/ARS, Fort Collins CO.

Objective: To review, test, and develop adjusted cryopreservation protocols for different cultivated genotypes of Pineapple and Citrus.

Approach: Pineapple material - In vitro plants of two commercial varieties (‘Perolera’ and ‘Primavera’) and one ornamental hybrid. Citrus material - Embryogenic callus of citrus obtained from aborted eggs of young plants of selected varieties; cell suspensions and seeds from Citrus sinensis and Citrus sunki. In vitro shoot cultures of pineapple and embryogenic cultures of citrus were initiated and bulked up in Brazil and transported to US as in vitro cultures by Dr. Fernanda Souza, who arrived on May/2012 at NCGRP to start her research work as visiting scientist from Embrapa Cassava and Fruits. Shoot tips of pineapple and somatic embryos and cell suspensions of citrus will be subjected to variations of existing protocols for cryopreservation based on different techniques like encapsulation-dehydration, vitrification, including droplet vitrification. These methods are routinely performed in the NCGRP laboratory and a large part of this research will be to transfer the cryopreservation knowledge and technology so that this research can continue in Brazil.

Results: Some preliminary assays have been established for pineapple to determine some critical points for designing experiments like sugar and PVS2 tolerance. The results showed a high sensitivity of pineapple shoot tips to high concentrations of sucrose. Two experiments based on a published protocol for pineapple (Martinez-Montero et al, 2002) were carried out using two explants sources: shoot tips and meristems from etiolated plants. Preliminary results showed no survival in liquid nitrogen reinforcing the need to develop a new approach for pineapple varieties.