2.3. Inoculum preparation, plant treatment and inoculation
procedures
Xcc inoculumwas prepared in nutrient broth and grown at 28 C
for 24 h to log phase. Bacterial suspension was centrifuged at
10,000 g for 20 min, re-suspended in phosphate buffer saline (PBS;
40 mM Na2HPO4 þ 25 mM KH2PO4), and adjusted to 104 cfu mL1
for attached leaf inoculations and 105 cfu mL1 for detached leaf
inoculations as previously described (Francis et al., 2010).
Nine-month-old ‘Pineapple’ sweet orange plants growing in 2.5-
L containers of a general purpose peat-based soil (Pro-Mix BX;
Premier Horticulture, Red Hill, PA) were maintained in a greenhouse
located at the Citrus Research and Education Center in Lake
Alfred, FL. Four weeks prior to the treatments, seedlings were cut
back to approximately 40 cm, and only one shoot per plant was
allowed to grow to approximately 20e30 cm in order to obtain 4e5
immature leaves (75% expanded) suitable for treatment and/or
inoculation.
2.3. Inoculum preparation, plant treatment and inoculationproceduresXcc inoculumwas prepared in nutrient broth and grown at 28 Cfor 24 h to log phase. Bacterial suspension was centrifuged at10,000 g for 20 min, re-suspended in phosphate buffer saline (PBS;40 mM Na2HPO4 þ 25 mM KH2PO4), and adjusted to 104 cfu mL1for attached leaf inoculations and 105 cfu mL1 for detached leafinoculations as previously described (Francis et al., 2010).Nine-month-old ‘Pineapple’ sweet orange plants growing in 2.5-L containers of a general purpose peat-based soil (Pro-Mix BX;Premier Horticulture, Red Hill, PA) were maintained in a greenhouselocated at the Citrus Research and Education Center in LakeAlfred, FL. Four weeks prior to the treatments, seedlings were cutback to approximately 40 cm, and only one shoot per plant wasallowed to grow to approximately 20e30 cm in order to obtain 4e5immature leaves (75% expanded) suitable for treatment and/orinoculation.
การแปล กรุณารอสักครู่..