Gene expression in barley upon aphid attack

Considering the fact that plants are unable to avoid their predators by walking, they use some other defense systems, like induction or repression of defense genes. A test carried out with barley attacked by the bird cherry-oat aphid (Rhopalosiphum padi), resulted in the hypothesis that contig 16360 (similar to ser/thr kinases) could possibly be connected to the resistance of barley against R. padi, and contig 6519 (similar to WIR 1A) with the susceptibility. Time course experiments confirmed that contig16360 and AJ250283 (much like BCI-4) are nearly induced just as, each, by 2 different aphids (R. padi and Metopolophium dirhodum). Genomic PCR was utilized to examine the hypothesis that when plants have the gene for contig 16360, they’re more likely to be immune to aphid attack, and when plants have the gene for contig 6519, they’re more susceptible. This experiment was carried out with Sixty nine barley lines: wild, commercial or breeding lines. Outcome was that the existence of WIR 1A gene doesn’t have correlation with the vulnerability, while presence of ser/thr kinase appears to be correlated with resistance.

Video: Aphid Attack

Various plants defense systems
Plant predators in insect kingdom
Problems caused by aphids on plants
Aphids: Diuraphis noxia, Metopolophium dirhodum, Rhopalosiphum padi
Background of the experiment, and studied genes 5
Aims of the project
Materials and methods
List of used primers
Plant growth
DNA extraction from Barley
Genomic PCR
Reverse Transcription-PCR (RT-PCR)
Real-time reverse PCR
Rapid Amplification of cDNA ends (RACE)
DNA and RNA visualisation after PCR
Genes (contig 16360 and AJ250283) regulation during aphid attack (M. dirhodum and R. padi)
Real-time reverse PCR analysis
Genomic PCR analysis
RACE experiment….

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Source: Sodertorn University

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