Showing papers by "Makio Takeda published in 2018"

Differential Proteomic Analysis of Date Palm Leaves Infested with the Red Palm Weevil (Coleoptera: Curculionidae)

Abstract: The red palm weevil, Rhynchophorus ferrugineus (Oliv.) (Coleoptera: Curculionidae), is a highly damaging pest of palm trees worldwide. The infestation is highly concealed in nature. Hence, a highly sensitive and reliable early detection technique needs to be applied in the field for identification and treatment of the infested date palms to curtail further infestation. We have recently reported the differential proteomic analysis of the date palm stem tissues associated with the red palm weevil infestation. In this study, we examine the response of date palm infested with red palm weevil based on the leaf proteome expression changes detected using two-dimensional differential gel electrophoresis (2D-DIGE) followed by Matrix-Assisted Laser Desorption/Ionization-Time-of-Flight (MALDI-TOF). We observed qualitative and quantitative proteome differences between the control and weevil-infested date palm samples. The red palm weevil infestation induced specific responses attributable to weevil feeding, relative to artificially wounded trees (which were used as a control). Differential proteomics led to the identification of 32 red palm weevil infestation-specific protein spots (P ≤ 0.05 having ≥ 1.5-fold modulation) that were further subjected to mass spectrometer analysis for identification and characterization. Proteins involved in plant stress and plant defense, photosynthesis, carbohydrate utilization, and protein degradation were affected in infested plants. The differentially expressed red palm weevil infestation-specific peptides can be used as biomarkers for the identification of early infestation with this insect in date palm trees. Moreover, our study demonstrates the potential use of proteomic strategies in diagnosing phyto-infestation caused by insect pests, diseases, and perhaps even for variety selection.

Molecular and functional characterization of the American cockroach, Periplaneta americana, Rab5: the first exopterygotan low molecular weight ovarian GTPase during oogenesis

Abstract: The small Rab GTPases are key regulators of membrane vesicle trafficking Ovaries of Periplaneta americana (Linnaeus) (Blattodea: Blattidae) have small molecular weight GTP/ATP-binding proteins during early and late vitellogenic periods of oogenesis However, the identification and characterization of the detected proteins have not been yet reported Herein, we cloned a cDNA encoding Rab5 from the American cockroach, P americana, ovaries (PamRab5) It comprises 796 bp, encoding a protein of 213 amino acid residues with a predicted molecular weight of 235 kDa PamRab5 exists as a single-copy gene in the P americana genome, as revealed by Southern blot analysis An approximate 26 kb ovarian mRNA was transcribed especially at high levels in the previtellogenic ovaries, detected by Northern blot analysis The muscle and head tissues also showed high levels of PamRab5 transcript PamRab5 protein was localized, via immunofluorescence labeling, to germline-derived cells of the oocytes, very early during oocyte differentiation Immunoblotting detected a ∼25 kDa signal as a membrane-associated form revealed after application of detergent in the extraction buffer, and 23 kDa as a cytosolic form consistent with the predicted molecular weight from amino acid sequence in different tissues including ovary, muscles and head The PamRab5 during late vitellogenic periods is required to regulate the endocytotic machinery during oogenesis in this cockroach This is the first report on Rab5 from a hemimetabolan, and presents an inaugural step in probing the molecular premises of insect oocyte endocytotic trafficking important for oogenesis and embryonic development