1a.Objectives (from AD-416):
The objective for this project will be to obtain the necessary data for foreign regulatory packages for the Plum pox virus resistant plum cultivar 'HoneySweet', including sequencing the 'HoneySweet' genome, identifying the location and structure of all transgene insertion events, and conducting fruit compositional studies. The objectives include the maintenance and increase in communications with foreign collaborators and regulatory agencies, and the support of regulatory submissions by foreign collaborators based on data developed through this agreement.
1b.Approach (from AD-416):
The 'HoneySweet' genome will be sequenced. Assembly will be performed using the peach genome as a template. Transgene insertion events will be identified via BLAST. Existing Prunus reference markers will be tested on 'HoneySweet' to refine the assembled physical map and determine the chromosome locations of transgene insertion events. Fruit compositional studies will be conducted and compared with controlled non-genetically engineered plums. Fruit of 'HoneySweet' and a number of non-GE controlled varieties will be collected in multiple years in a design suitable for statistical analyses of composition. Samples will be analyzed by a certified lab. Meetings between the U.S. and foreign collaborators will be for the purpose of sharing data and for the planning of regulatory package submissions, which will be supported through this agreement.
Total messenger RNA and small RNAs from ‘HoneySweet’ grown in two locations in Europe and one location in the U.S. was sequenced using Next Generation sequencing. RNA from the plum cultivar, ‘Stanley’, from the same locations was also sequenced. Comparisons of the sequences were made with no significant differences found in the expression of messenger RNA from the transgenic ‘HoneySweet’. The transgene expression was found to be at a low level.