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.
3. Progress Report:
The 'HoneySweet' genome has been sequenced. The nature and location of the insert is verified and as predicted through previous molecular analyses. The absence of DNA from outside of the T-DNA borders was confirmed. The genetic structure in terms of genes and gene order between plum (Prunus domestica) and peach (P. persica) is similar, and information from the peach genome can be readily transferred to plum. Studies of the small RNAs produced by 'HoneySweet' plum have been initiated based on the sequence of the insert and the genome sequence of 'HoneySweet'. The small RNA profile is being compared with non-GE plums.