TABLE 1.
Percentages of “Ca. Liberibacter asiaticus”-exposed D. citri samples that contained “Ca. Liberibacter asiaticus” and Wolbachia
| Samplea | Stage | Bacterium | % with Ct |
Mean copy no.b | |
|---|---|---|---|---|---|
| <35 | >35 | ||||
| Whole body | Nymph (5th instar) | “Ca. Liberibacter asiaticus” | 70.0 | 30.0 | 744.3 |
| Wolbachia | 100.0 | 0.0 | 20,115.6 | ||
| Adult | “Ca. Liberibacter asiaticus” | 58.6 | 41.4 | 539.0 | |
| Wolbachia | 100.0 | 0.0 | 14,829.8 | ||
| Gut | Nymph (5th instar) | “Ca. Liberibacter asiaticus” | 60.0 | 40.0 | 331.1 |
| Wolbachia | 100.0 | 0.0 | 118,160.2 | ||
| Adult | “Ca. Liberibacter asiaticus” | 86.2 | 13.8 | 1,846.1 | |
| Wolbachia | 100.0 | 0.0 | 723,056.0 | ||
Each sample was tested for both “Ca. Liberibacter asiaticus” and Wolbachia titers. For each life stage, 30 samples were used, and each sample had three technical replicates, which were averaged for a final Ct value. The samples with an average technical-replicate Ct value of <35 contributed to the percent infection rate (e.g., for the whole-body nymphs tested, 21/30 samples had Ct values of <35, for a 70% “Ca. Liberibacter asiaticus” infection rate). The individuals used for guts and whole bodies were taken from separate generations of the same colonies. Exposed nymphs and adults were fed on “Ca. Liberibacter asiaticus”-infected citrus trees for one or more generations.
The mean copy number is the average of individual sample copy numbers with Ct values of <35 using qPCR. Cycle threshold values were converted to copy numbers using a standard curve.