Attention! iBeetle is a first pass screen. Hence, the documented phenotypes need to be confirmed by independent experiments using non-overlapping dsRNA fragments! Phenotypes with a penetrance > 50% are frequently reproduced. See Schmitt-Engel et al. 2015 for details on reproducibility.
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| iB_03687 | iB fragment
The sequence from the iB fragment is matched against the genes from the official gene set. The matche(s) is/are visualizied in a genome browser. |
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Pupal injection procedure
Day 0: 10 female pupae of the pBA19 strain (muscle enhancer trap line) were injected with dsRNA.
3 days post injection (dpi): Hatch control: Pupal and adult lethality as well as metamorphosis defects (molting, eclosion) were documented. For mating, 4 males of the black strain were added.
9 dpi: First egg-lay was collected and incubated for cuticle analysis. Adult lethality and egg production (reduced/ no egg-lay) was documented.
11 dpi: Second egg-lay was collected and incubated for embryonic muscle analysis. Adult lethality and egg production (reduced/ no egg-lay) were documented.
Note: The adult morphology was not analyzed systematically at 3/9/11 dpi. Only obvious phenotypes, visible without magnification, have been annotated.
13 dpi: Egg productivity and Ovary analysis
The percentage of hatched larvae was documented and not hatched larvae/ eggs were embedded for cuticle analysis (15 dpi).
In case of a reduction of egg production, 4 injected females were dissected to analyze the gross morphology of the ovaries.
14 dpi: Analysis of embryonic musculature and early embryonic development
Offspring of the injected females (hatched and not hatched larvae/ eggs) were analysed for embryonic lethality and muscle defects.
15 dpi: Analysis of larval instar 1 cuticle
Offspring of injected females were analysed and cuticle phenotypes were annotated.
22 dpi: Stink gland analysis
Documentation of defects in abdominal and thoracic stink glands (colour, size, content) of the injected femals.
Terms used in the pupal injection procedure
- larval pantagmatic defects
- At least two tagmata (head, thorax, abdomen or terminus) show similar/ comparable defects.
- empty eggs
- Just empty shells are visible, no cuticle has been developed.
- strong defects
- L1 cuticle larvae which show severe defects. These strong defects are subdivided into three categories
- strongly fragmented (cuticle crumbs)
- No segments or appendages are identifiable, only residues of cuticle and bristles can be identified.
- cuticle remnants (with unidentifiable segments)
- Segments or parts of appendages are recognizable, but identity is not clear.
- cuticle remnants (with some identifiable segments)
- At least one segment or appendage is clearly regonizable.
- number of eggs on slide
- affected embryos usually (not always) do not hatch. Only the not hatched were used for cuticle analysis.
For the embryonic muscle analysis the hatched as well as the not hatched were analysed. - total number of affected eggs/embryos/larvae
- Summary of all different phenotypes/ phenotypic series which were annotated.
- inside-out
- The L1 larva shows a partial or complete inversion., e.g. bristles, appendages or parts of the abdominal segments are inverted into the interior of the cuticle.
- starved
- Dissected females show a strong resorption of fatbody predominately in the abdomen and the oogenesis is blocked. These are considered as eventually lethal phenotypes and the blocked oogenesis probably a secondary defect due to starvation.
- eclosion not fulfilled
- The emergence of the adult from the pupa stage is interrupted. This phenotype shows pupal as well as adult features.
Lethality 11 days after pupal injection: 10.0% (includes death as pupa, adult)
eclosion not fulfilled (with pupal and adult features) - Number of animals showing the phenotype: 1 (3 dpi)Larval injection procedure
Day 0: 10 female L5-6 larvae of the D17Xhom strain (females express RFP in the eyes and in the CNS; pupae express GFP in the thorax) were injected with dsRNA.
11 days post injection (dpi): Pupal morphology Documentation of larval/ pupal death and defects of metamorphosis (eclosion, delayed development). Screening for alterations of pupal head and appendages, pupal thorax, pupal thoracic musculature, pupal legs, pupal wings, pupal abdomen and pupal genital lobes.
16 dpi: Adult morphology Documentation of larval/ pupal and adult death and defects of metamorphosis (eclosion, delayed development). Screening for alterations of adult head and appendages, adult thorax, adult elytra and hindwings, adult legs and cuticle properties. For mating 4 males of the black strain were added.
19 dpi: Sieving and egg-lay Documentation of adult lethality and enduring metamorphosis defects.
22 dpi: Ovary analysis (Fertility) In case of a reduction of egg production 4 females were dissected and the gross morphology of the ovaries were analysed.
41 dpi: Fertility and stink gland analysis Documentation of defects in abdominal and thoracic stink glands development (colour, size, content) of the injected females. Check for fertility of injected larvae (no or decreased offspring).
Terms used in the larval injection procedure
- pupal/ adult pantagmatic defects
- At least two tagmata (head, thorax, abdomen or terminus) show similar/ comparable defects.
- pupal molt not fulfilled
- The hatching from the larva to the pupa is interrupted. Either it dies as prepupa or the phenotype shows larval as well as pupal features.
- eclosion not fulfilled
- The emergence of the adult from the pupa stage is interrupted. This phenotype shows pupal as well as adult features.
- starved
- Dissected females show a strong resorption of fatbody predominately in the abdomen and the oogenesis is blocked. These are considered as eventually lethal phenotypes and the blocked oogenesis probably a secondary defect due to starvation.
Lethalities 11 days after larval injection: 10.0% (includes death as larva, prepupa, pupa)
Lethalities 22 days after larval injection: 40.0% (includes death as larva, prepupa, pupa, adult)
eclosion not fulfilled (died as pupa) - Number of animals showing the phenotype: 3 (16 dpi)
- >TC004084 RA
MAVIGYGET ADSQLGWDC GGTLISELY VLTAAHCLE SRELGPSQL VRFGTTHLD EPDPDLQER VVVARIPHP DYKPPLKAN DIGLIKLEE PVEFTPHVR PACLNTADI NPGRKALAS GFGKLSYDA ETGSKNLMK VLLNVYPNN RCSKAIREQ IKDTMLCAG HLEGGKDTC QGDSGGPLQ IVLEKPYCM YSVIGVTSF GKFCGFANA PAIYTKISA YISWIESIV WK
- >TC004084 RB
MQVMHGNLQ LPSNISSTK CEEYAKAVY VQTISPVLS LNAKTNNVS ECGIVSVPL IIGGTAATE KEFPHMAVI GYGETADSQ LGWDCGGTL ISELYVLTA AHCLESREL GPSQLVRFG TTHLDEPDP DLQERVVVA RIPHPDYKP PLKANDIGL IKLEEPVEF TPHVRPACL NTADINPGR KALASGFGK LSYDAETGS KNLMKVLLN VYPNNRCSK AIREQIKDT MLCAGHLEG GKDTCQGDS GGPLQIVLE KPYCMYSVI GVTSFGKFC GFANAPAIY TKISAYISW IESIVWK
- >TC004084 RA
atggctgtg attggatat ggggaaaca gcagattct cagctcgga tgggattgt ggaggcact ttaattagt gaattatat gttttaaca gctgctcat tgcttggaa agtcgagaa ttaggacca tctcaactt gtccgcttt ggtactaca catttagat gagccggat cccgatttg caagaacgt gtagttgtt gcaagaatc cctcatcca gactataaa ccaccgcta aaagccaac gatataggt ttaattaaa ctagaagaa cccgtagaa ttcacacca catgtaaga cctgcttgt ttaaatacc gctgatata aatcctgga cgaaaagca ctagcatca gggtttggg aaattgtcg tacgatgcg gaaaccggg agtaaaaac ttaatgaaa gtattactc aatgtttat cccaacaat cgatgctcg aaagcaata cgagaacag ataaaggac acaatgctg tgtgctgga catttggaa ggaggcaaa gatacttgt cagggagac tcaggaggt ccattgcag attgtttta gaaaaacca tactgtatg tatagtgtt attggagtt acttccttt gggaaattt tgtggtttt gctaatgca ccagccatt tatacaaaa atttccgct tatatttct tggattgaa agtattgta tggaaataa
- >TC004084 RB
atgcaagtg atgcacgga aacttgcaa cttcctagt aatatttca tccacgaaa tgtgaggaa tatgcaaaa gcagtttat gttcaaact atcagtcca gtgctgtct ctaaacgcc aaaacaaat aatgtatcc gaatgtgga atcgtgtca gtgccttta ataataggc ggaacagct gcaaccgaa aaagaattt cctcacatg gctgtgatt ggatatggg gaaacagca gattctcag ctcggatgg gattgtgga ggcacttta attagtgaa ttatatgtt ttaacagct gctcattgc ttggaaagt cgagaatta ggaccatct caacttgtc cgctttggt actacacat ttagatgag ccggatccc gatttgcaa gaacgtgta gttgttgca agaatccct catccagac tataaacca ccgctaaaa gccaacgat ataggttta attaaacta gaagaaccc gtagaattc acaccacat gtaagacct gcttgttta aataccgct gatataaat cctggacga aaagcacta gcatcaggg tttgggaaa ttgtcgtac gatgcggaa accgggagt aaaaactta atgaaagta ttactcaat gtttatccc aacaatcga tgctcgaaa gcaatacga gaacagata aaggacaca atgctgtgt gctggacat ttggaagga ggcaaagat acttgtcag ggagactca ggaggtcca ttgcagatt gttttagaa aaaccatac tgtatgtat agtgttatt ggagttact tcctttggg aaattttgt ggttttgct aatgcacca gccatttat acaaaaatt tccgcttat atttcttgg attgaaagt attgtatgg aaataa
- >TC004084 RA
aaacgccaa aacaaataa tgtatccga atgtggaat cgtgtcagt gcctttaat aataggcgg aacagctgc aaccgaaaa agaatttcc tcacatggc tgtgattgg atatgggga aacagcaga ttctcagct cggatggga ttgtggagg cactttaat tagtgaatt atatgtttt aacagctgc tcattgctt ggaaagtcg agaattagg accatctca acttgtccg ctttggtac tacacattt agatgagcc ggatcccga tttgcaaga acgtgtagt tgttgcaag aatccctca tccagacta taaaccacc gctaaaagc caacgatat aggtttaat taaactaga agaacccgt agaattcac accacatgt aagacctgc ttgtttaaa taccgctga tataaatcc tggacgaaa agcactagc atcagggtt tgggaaatt gtcgtacga tgcggaaac cgggagtaa aaacttaat gaaagtatt actcaatgt ttatcccaa caatcgatg ctcgaaagc aatacgaga acagataaa ggacacaat gctgtgtgc tggacattt ggaaggagg caaagatac ttgtcaggg agactcagg aggtccatt gcagattgt tttagaaaa accatactg tatgtatag tgttattgg agttacttc ctttgggaa attttgtgg ttttgctaa tgcaccagc catttatac aaaaatttc cgcttatat ttcttggat tgaaagtat tgtatggaa ataaaaata aatgtatta tataata
- >TC004084 RB
gagttttcc caatatctc attatgcaa gtgatgcac ggaaacttg caacttcct agtaatatt tcatccacg aaatgtgag gaatatgca aaagcagtt tatgttcaa actatcagt ccagtgctg tctctaaac gccaaaaca aataatgta tccgaatgt ggaatcgtg tcagtgcct ttaataata ggcggaaca gctgcaacc gaaaaagaa tttcctcac atggctgtg attggatat ggggaaaca gcagattct cagctcgga tgggattgt ggaggcact ttaattagt gaattatat gttttaaca gctgctcat tgcttggaa agtcgagaa ttaggacca tctcaactt gtccgcttt ggtactaca catttagat gagccggat cccgatttg caagaacgt gtagttgtt gcaagaatc cctcatcca gactataaa ccaccgcta aaagccaac gatataggt ttaattaaa ctagaagaa cccgtagaa ttcacacca catgtaaga cctgcttgt ttaaatacc gctgatata aatcctgga cgaaaagca ctagcatca gggtttggg aaattgtcg tacgatgcg gaaaccggg agtaaaaac ttaatgaaa gtattactc aatgtttat cccaacaat cgatgctcg aaagcaata cgagaacag ataaaggac acaatgctg tgtgctgga catttggaa ggaggcaaa gatacttgt cagggagac tcaggaggt ccattgcag attgtttta gaaaaacca tactgtatg tatagtgtt attggagtt acttccttt gggaaattt tgtggtttt gctaatgca ccagccatt tatacaaaa atttccgct tatatttct tggattgaa agtattgta tggaaataa aaataaatg tattatata ata
