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Identification of Proteins Modulated in the Date Palm Stem Infested with Red Palm Weevil (Rhynchophorus ferrugineus Oliv.) Using Two Dimensional Differential Gel Electrophoresis and Mass Spectrometry.

Rasool KG, Khan MA, Aldawood AS, Tufail M, Mukhtar M, Takeda M - Int J Mol Sci (2015)

Bottom Line: A constant unique pattern of differential expression in infested (I), wounded (W) stem samples compared to control (C) was observed.All 32 differentially expressed spots were subjected to MALDI-TOF analysis for their identification and we were able to match 21 proteins in the already existing databases.Relatively significant modulated expression pattern of a number of peptides in infested plants predicts the possibility of developing a quick and reliable molecular methodology for detecting plants infested with date palm.

View Article: PubMed Central - PubMed

Affiliation: Department of Plant Protection, College of Food and Agriculture Sciences, King Saud University, Riyadh 11451, Saudi Arabia. krasool@ksu.edu.sa.

ABSTRACT
A state of the art proteomic methodology using Matrix Assisted Laser Desorption/Ionization-Time of Flight (MALDI TOF) has been employed to characterize peptides modulated in the date palm stem subsequent to infestation with red palm weevil (RPW). Our analyses revealed 32 differentially expressed peptides associated with RPW infestation in date palm stem. To identify RPW infestation associated peptides (I), artificially wounded plants (W) were used as additional control beside uninfested plants, a conventional control (C). A constant unique pattern of differential expression in infested (I), wounded (W) stem samples compared to control (C) was observed. The upregulated proteins showed relative fold intensity in order of I > W and downregulated spots trend as W > I, a quite interesting pattern. This study also reveals that artificially wounding of date palm stem affects almost the same proteins as infestation; however, relative intensity is quite lower than in infested samples both in up and downregulated spots. All 32 differentially expressed spots were subjected to MALDI-TOF analysis for their identification and we were able to match 21 proteins in the already existing databases. Relatively significant modulated expression pattern of a number of peptides in infested plants predicts the possibility of developing a quick and reliable molecular methodology for detecting plants infested with date palm.

No MeSH data available.


Related in: MedlinePlus

2D-DIGE images of date palm proteins. The protein sample of Infested-1, Control-3 and internal standard (pooled of all the samples) are individually labeled with Cy dyes, mixed together and separated by 2D-DIGE followed by image scanning. (A) Image of date palm infested sample and labeled with Cy3 dye; (B) Image of date palm control sample labeled with Cy5 dye; (C) Image of date palm sample pooled from all and labeled with Cy2 dye; and (D) Overlay gel of infested, and control samples along with internal standard.
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ijms-16-19326-f002: 2D-DIGE images of date palm proteins. The protein sample of Infested-1, Control-3 and internal standard (pooled of all the samples) are individually labeled with Cy dyes, mixed together and separated by 2D-DIGE followed by image scanning. (A) Image of date palm infested sample and labeled with Cy3 dye; (B) Image of date palm control sample labeled with Cy5 dye; (C) Image of date palm sample pooled from all and labeled with Cy2 dye; and (D) Overlay gel of infested, and control samples along with internal standard.

Mentions: After labeling with Cy dyes, two samples were mixed with different combinations along with internal standard and electrophoresed on the same gel except one gel contained a single sample with internal standard. The representative gels of 2D-DIGE after scanning with a fluorescence gel scanner, Typhoon imager (Trio) (GE Healthcare), are shown in Figure 2.


Identification of Proteins Modulated in the Date Palm Stem Infested with Red Palm Weevil (Rhynchophorus ferrugineus Oliv.) Using Two Dimensional Differential Gel Electrophoresis and Mass Spectrometry.

Rasool KG, Khan MA, Aldawood AS, Tufail M, Mukhtar M, Takeda M - Int J Mol Sci (2015)

2D-DIGE images of date palm proteins. The protein sample of Infested-1, Control-3 and internal standard (pooled of all the samples) are individually labeled with Cy dyes, mixed together and separated by 2D-DIGE followed by image scanning. (A) Image of date palm infested sample and labeled with Cy3 dye; (B) Image of date palm control sample labeled with Cy5 dye; (C) Image of date palm sample pooled from all and labeled with Cy2 dye; and (D) Overlay gel of infested, and control samples along with internal standard.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4581299&req=5

ijms-16-19326-f002: 2D-DIGE images of date palm proteins. The protein sample of Infested-1, Control-3 and internal standard (pooled of all the samples) are individually labeled with Cy dyes, mixed together and separated by 2D-DIGE followed by image scanning. (A) Image of date palm infested sample and labeled with Cy3 dye; (B) Image of date palm control sample labeled with Cy5 dye; (C) Image of date palm sample pooled from all and labeled with Cy2 dye; and (D) Overlay gel of infested, and control samples along with internal standard.
Mentions: After labeling with Cy dyes, two samples were mixed with different combinations along with internal standard and electrophoresed on the same gel except one gel contained a single sample with internal standard. The representative gels of 2D-DIGE after scanning with a fluorescence gel scanner, Typhoon imager (Trio) (GE Healthcare), are shown in Figure 2.

Bottom Line: A constant unique pattern of differential expression in infested (I), wounded (W) stem samples compared to control (C) was observed.All 32 differentially expressed spots were subjected to MALDI-TOF analysis for their identification and we were able to match 21 proteins in the already existing databases.Relatively significant modulated expression pattern of a number of peptides in infested plants predicts the possibility of developing a quick and reliable molecular methodology for detecting plants infested with date palm.

View Article: PubMed Central - PubMed

Affiliation: Department of Plant Protection, College of Food and Agriculture Sciences, King Saud University, Riyadh 11451, Saudi Arabia. krasool@ksu.edu.sa.

ABSTRACT
A state of the art proteomic methodology using Matrix Assisted Laser Desorption/Ionization-Time of Flight (MALDI TOF) has been employed to characterize peptides modulated in the date palm stem subsequent to infestation with red palm weevil (RPW). Our analyses revealed 32 differentially expressed peptides associated with RPW infestation in date palm stem. To identify RPW infestation associated peptides (I), artificially wounded plants (W) were used as additional control beside uninfested plants, a conventional control (C). A constant unique pattern of differential expression in infested (I), wounded (W) stem samples compared to control (C) was observed. The upregulated proteins showed relative fold intensity in order of I > W and downregulated spots trend as W > I, a quite interesting pattern. This study also reveals that artificially wounding of date palm stem affects almost the same proteins as infestation; however, relative intensity is quite lower than in infested samples both in up and downregulated spots. All 32 differentially expressed spots were subjected to MALDI-TOF analysis for their identification and we were able to match 21 proteins in the already existing databases. Relatively significant modulated expression pattern of a number of peptides in infested plants predicts the possibility of developing a quick and reliable molecular methodology for detecting plants infested with date palm.

No MeSH data available.


Related in: MedlinePlus