Optimization of protein extraction for differential proteomic analysis of the
date palm infested with red palm weevil
Khawaja Ghulam Rasool1, Abdulrahman S. Aldawood1, Muhammad Tufail1 and Makio
Takeda2
1
Department of Plant Protection, King Saudi University, Riyadh, Saudi Arabia
2
Graduate School of Agricultural Science, Kobe University, Japan
Abstract:
The Kingdom of Saudi Arabia is one of the major producers of fine quality dates and ranks
second in global dates production. This important fruit crop is under threat due to highly invasive
pest, the red palm weevil (RPW) (Rhynchophorus ferrugineus). Failure in early detection of RPW
infestation is the major obstacle in its successful management. In present study, we plan to identify
the differentially expressed peptides of date palm possibly associated with red palm weevil
infestation.
Five protein extraction protocols with some modifications were used to extract protein
from date palm leaves and stem. The objective of protein extraction protocol optimization was to
select the best one with good protein yield in short time period. Therefore, yield and protein
profiling were compared among the tested protocols. We observed that protocol with phenol/SDS
extraction gave higher yield with reduced number of steps.
Mini 2DE gels were used to assess reproducibility of protein profiling of infested and
control protein samples. Both infested and control samples showed abundant proteins of different
molecular weight/ sizes. Some protein spots were identified revealing variability in the intensity
or relative abundance. The comparison of infested and control gels revealed that some protein
spots were relatively up regulated. Also, some additional spots were identified in the infested
samples. However, our future experiments with DIGE and Maldi TOF TOF will confirm the
integrity of these differentially expressed peptides.