Young Lso-free adult psyllids (less than 7 days old, a mix of males and females) were transferred to LsoA- or LsoB-infected tomato plants. To minimize the effect of differences in Lso titer within infected plants19 (link), groups of over 60 adult psyllids were caged on leaves at the same level of the tomato plants using organza bags. Every 2 days, 50 insects from a same group were collected in order to obtain 2-, 4-, 6-, 8-, 10-, 12-, 14-, and 16-day Lso-exposed psyllids. The groups of 50 psyllids were randomly caged on different plants. The experiments were stopped after 16 days of AAP because adult tomato psyllids live on average 1 month and high mortality of psyllids started to be observed.
Guts from the LsoA- or LsoB-exposed psyllids were dissected under the dissecting microscope36 (link). DNA from pools of 50 guts was purified following the protocol of blood/tissue DNA extraction kit (Qiagen, Hilden, Germany); each pool was used as an individual template for qPCR analysis. Thus, each pool of 50 guts represented one replicate, and there were three replicates for each combination of exposure time point and haplotype. Each replicate was obtained by using independent LsoA- or LsoB-infected plants as Lso inoculum. All the experiments were conducted at 24 ± 1 °C.
Guts from the LsoA- or LsoB-exposed psyllids were dissected under the dissecting microscope36 (link). DNA from pools of 50 guts was purified following the protocol of blood/tissue DNA extraction kit (Qiagen, Hilden, Germany); each pool was used as an individual template for qPCR analysis. Thus, each pool of 50 guts represented one replicate, and there were three replicates for each combination of exposure time point and haplotype. Each replicate was obtained by using independent LsoA- or LsoB-infected plants as Lso inoculum. All the experiments were conducted at 24 ± 1 °C.
Free full text: Click here
