Seed decontamination: 1. Develop guidelines to prevent seed contamination by canker. Part 1. Surface disinfection of seeds. Objective is to develop economical and practical cleaning methods for removing Xcc without loss of seed viability. Progress: Thirty freshly squeezed seeds of Kuharske were plated on LB plates. Concurrently rinsate of seeds rinsed in sterile water had serial dilutions plated on media plates viz. V8, PDA, LB and Dox. LB plates showed numerous distinct colonies, whereas the rest showed bacterial smears. Bacterial colonies on LB plates resembled X. citri and other unknown bacteria. Similar results occurred on LB plates plated with freshly squeezed intact seeds. Identity of these bacteria is unknown, but appear to be common environmental bacteria. Identification requires DNA sequencing, planned in the future. Seeds treated with different concentrations of chlorox and PPM appeared to display higher seed germination rates, to be investigated in the future. Part 2. Evaluating the 2006 CHRP decontamination procedure. Progress: Experiments are in progress along with experiments using sodium hypochlorite and PPM for decontamination. Part 3. Development of diagnostic methods for on-site canker detection. Progress: Experiments are ongoing. Cutting and tissue cultivation In October propagation beds with bottom heat were constructed. Sides and separators were installed on 3 benches, producing 12 independent beds. Beds were partially filled with perlite and heating cables installed with independent temperature controls. On 14 October, 4 schedules of acclimation of tissue cultured rootstocks were applied to C-35, C-54 sour orange, US-812, SW-13 and KC-13 root stocks. Any roots were removed and stems transplanted into peat:perlite in seedling trays. Plants were harvested on 11/16. Success was >92% for all rootstocks except sour orange (82%). The next set of tissue culture plants were planted on 12/14, with additional plants placed in 4 of the heated sections; with the same levels of light and heat exclusion. Changes in relative humidity and light levels were the same reported previously. The gap in the sequence was due to insufficient numbers of plantlets the first delivered, with delays in obtaining the remainder. Due to technical issues, humidity levels were insufficient the first week and many plantlets died. Plants in heated benches faired better due to higher humidity. These plants will be harvested in mid-February. Cutting propagation continued with single node cuttings of Kuharski initiated on 10/14, 11/3 and 12/1. Harvest of cuttings have continued to be every 6 weeks, with harvest occurring on 10/14, 11/3, 12/1. Auxin concentrations of 4000 and 7500 ppm have been consistent. Most woody plants require higher auxin concentrations during the winter months for good rooting. Rooted cuttings of the HLB resistant lines were transplanted in 5� tree pots the week before Christmas. Any cutting that had good root systems were transplant, independent of shoot growth. Less than 10 cutting failed to produce roots. Even plants with no additional shoot growth had abundant root growth at transplanting. These plants were placed in the production side of the greenhouse due to insufficient space in the propagation bay.