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Characterization of Insertion Sites in Rainbow Papaya, the First Commercialized Transgenic Fruit Crop |
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| Authors: | Jon Y Suzuki Savarni Tripathi Gustavo A Fermín Fuh-Jyh Jan Shaobin Hou Jimmy H Saw Christine M Ackerman Qingyi Yu Michael C Schatz Karen Y Pitz Marcela Yépes Maureen M M Fitch Richard M Manshardt Jerry L Slightom Stephen A Ferreira Steven L Salzberg Maqsudul Alam Ray Ming Paul H Moore Dennis Gonsalves |
| Institution: | 1. USDA-ARS Pacific Basin Agricultural Research Center, P.O. Box 4459, 64 Nowelo St., Hilo, HI, 96720, USA 7. Plant and Environmental Protection Sciences, College of Tropical Agriculture and Human Resources, University of Hawaii, Honolulu, HI, 96822, USA 2. Centro Jardín Botánico, Universidad de los Andes, Mérida, Venezuela 3. Department of Plant Pathology, National Chung Hsing University, Taichung, Taiwan, Republic of China 4. Advanced Studies in Genomics, Proteomics and Bioinformatics, University of Hawaii, Honolulu, HI, 96822, USA 11. Department of Microbiology, University of Hawaii, Honolulu, HI, 96822, USA 5. Hawaii Agricultural Research Center, Aiea, Honolulu, HI, 96701, USA 6. Center for Bioinformatics and Computational Biology, University of Maryland, College Park, MD, 20742, USA 8. Department of Plant Pathology, Cornell University, Geneva, New York, 14456, USA 9. Department of Tropical Plant and Soil Sciences, College of Tropical Agriculture and Human Resources, University of Hawaii, Honolulu, HI, 96822, USA 10. AureoGen Biosciences, Kalamazoo, MI, 49009, USA 12. Department of Plant Biology, University of Illinois at Urbana-Champaign, Urbana, IL, 61801, USA |
| Abstract: | Inserts and insert sites in transgenic, papaya ringspot virus (PRSV)-resistant commercial papaya Rainbow and SunUp, were characterized as part of a petition to Japan to allow import of fresh fruit of these cultivars from the U.S. and to provide data for a larger study aimed at understanding the global impact of DNA transformation on whole genome structure. The number and types of inserts were determined by Southern analysis using probes spanning the entire transformation plasmid and their sequences determined from corresponding clones or sequence reads from the whole-genome shotgun (WGS) sequence of SunUp papaya. All the functional transgenes, coding for the PRSV coat protein (CP), neophosphotransferase (nptII) and β-glucuronidase (uidA) were found in a single 9,789 basepair (bp) insert. Only two other inserts, one consisting of a 290 bp nonfunctional fragment of the nptII gene and a 1,533 bp plasmid-derived fragment containing a nonfunctional 222 bp segment of the tetA gene were detected in Rainbow and SunUp. Detection of the same three inserts in samples representing transgenic generations five to eight (R5 to R8) suggests that the three inserts are stably inherited. Five out of the six genomic DNA segments flanking the three inserts were nuclear plastid sequences (nupts). From the biosafety standpoint, no changes to endogenous gene function based on sequence structure of the transformation plasmid DNA insertion sites could be determined and no allergenic or toxic proteins were predicted from analysis of the insertion site and flanking genomic DNA. |
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