2006 Annual Report 4d.Progress report. This report documents research conducted under a Specific Cooperative Agreement between ARS and the University of Hawaii at Manoa. Additional details of research can be found in the report for the associated in-house CRIS 5320-22430-021-00D, Detection, Control and Area wide Management of Fruit Flies. Agrobacterium-mediated transformation of pineapple for nematode resistance and flowering control. Two transformation systems were developed for transformation of pineapple for nematode resistance and flowering control. In both cases, the low acid pineapple variety MD-1 was used as source plants. The construct for nematode resistance was a modified rice cystatin (ubi9-d86), 35S-GUS, 35S-Hpt (hygromycin resistance). The Agrobacterium was AGL0 and stem segments and leaf bases were the plant tissues infected; regeneration was via an adventitious bud pathway. For nematode resistance, a total of 22 PCR positive transgenic lines were produced. Over 300 individuals of line 101 are in greenhouse for nematode bioassays. All other lines have gone through at least 4 rounds of selection in hygromycin. All lines except for two have been transferred to rooting media containing hygromycin. For flowering control, the construct for flowering control was ACC antisense (ubi9), 35S-Hpt (hygromycin resistance). The promoters are constitutive. Agrobacterium AGL0 and stem segments and leaf bases were the plant tissues infected; regeneration was via an adventitious bud pathway. A total of 7 PCR positive transgenic lines were produced. Two additional lines have completed 3 rounds of testing and require further testing. Line 51 (over 300 individual plants) is being tested in greenhouse for bioassay of flowering control. Other lines have gone through at least 4 rounds of selection in hygromycin and are being multiplied in media with hygromycin. Using adventitious bud pathway, an Agrobacterium-mediated transformation method has been developed for pineapple. The method is highly repeatable and the transformation efficiency is about 1 transgenic line per thousand pieces of tissue used.