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Series GSE6363

Query DataSets for GSE6363

Status

Public on Nov 28, 2006

Title

Effect of day-length shortening on gene expression on pea aphid heads

Organism

Acyrthosiphon pisum

Experiment type

Expression profiling by array

Summary

Seasonal photoperiodism is responsable in aphid for the switch from asexual to sexual reproduction. In order to identify genes regulating the photoperiodic response, a cDNA microarray from the pea aphid was used to compare RNA populations from short- and long-day reared aphids. Analyses revealed that 64 different transcripts were significantly regulated, with a strong biological signature for cuticular proteins and proteins involved in cellular signalling or signal transduction. Quantitative PCR experiments performed on 5 transcripts confirmed microarray results. Complementary experiments eliminated moulting and circadian rhythms as putative confounding effects.
Keywords: Stress response, Environmental cue

Overall design

Two rearing conditions were compared: 16 h of light (“long-day” or LD) that maintained parthenogenesis, and 12 h of light (“short-day” or SD) that induced sexual forms after two generations. To initiate the experiment two groups of clonal third instar larvae (L3) were placed either at SD or LD photoperiod. This corresponds to generation G0. L3-G0 aphids reached adulthood (wingless adults or WA) and produced offspring (G1) that were transferred onto new plants and kept under LD or SD conditions. A batch of 50 L3-G1 aphids was collected from each condition in the middle of the photophase (15:00) and immediately frozen in liquid nitrogen. Triplicates were performed. Thus, there were 6 batches of aphids: 3 reared under LD and 3 reared under SD. Heads were cut and antennae discarded. RNA was extracted from the 6 batches. Aliquot of the 3 RNA extracts form LD batches were mix to compose the REFERENCE batch. Then each RNA extract was hybridized in competition with the RNA reference: RNA from SD (replicate1) hybridized against the Reference, RNA from LD (replicate 1) hybridized against the Reference, RNA from LD, RNA from SD (replicate 2) hybridized against the Reference, RNA from LD, and so on. This is 6 arrays. A dye swap was introduced, with the Reference labelled either with Cy3 or with Cy5. The experimental design is thus 12 arrays, which corresponds to the 12 described sample of that series.

Contributor(s)

LE TRIONNAIRE G, SABATER-MUNOZ B, BENEDETTO A, BONHOMME J, JAUBERT S, PRUNIER-LETERME N, MARTINEZ-TORRES D, SIMON J, TAGU D

Citation(s)

17785197

Submission date

Nov 24, 2006

Last update date

Mar 16, 2012

Contact name

Denis TAGU

E-mail(s)

denis.tagu@rennes.inra.fr