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_02784 | 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)
adult lethality - Number of animals showing the phenotype: 1 (9 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: -1.0% (includes death as larva, prepupa, pupa)
Lethalities 22 days after larval injection: -1.0% (includes death as larva, prepupa, pupa, adult)
- >TC010933 RA
MKFLAFFLL CVPHVLSLP KGSRIVGGN QARGGQFPF VAAIYVTRS GGTYFCGGV LLSNQWVLT AGQCVDGAT SFRVQLGSN ILEGTDLNR VTLATATYT LHPRYNSET LENDLGLIK LRLPIEFNQ YISVITLPS SFTPDYVSV TAVGWGQVD DFSTGLSNN LVSVTLTTI SHDECQLTY GPQITHDML CAAGNFNEG ICNGDMGGA LIQTIGGTA VHVGIASFR SHNGCETTD PSGFTRTIA YVDWIRNFV ALFSLFTLF SSPVYSNIA GRIIGGSDA KAGEFPFTA AIYIHTSSG QYFCAGALL SREWILTGA QCVDGALQF TIVLGSNNL QDSNDPNRV KVATEEYYL YPKYDAFTL QHDIALIKL RLPIEYSDY IQPILLPTA ELVDYSVVV ALGWGQTSD EDAGNVSDL HRVSITVLS NQECRNTYG TQIIEEMVC AEGNYNQGT CHGDSGGPL IQYVYGQSA THVGVSSFF SGNGCESTD PSGFTRTFP YVEWIRNVT QI
- >TC010933 RA
atgaaattc ctcgcattc tttttgctg tgtgtacca cacgttttg agtttgcct aaagggtct cgaattgtt ggaggaaac caggccaga ggtggtcag tttcccttt gtggcggca atttatgta acaaggtct ggaggtact tacttctgt ggaggtgtt ttgcttagt aatcagtgg gttctcacg gctggacaa tgcgtagat ggagccact tcgtttcga gttcaactt ggctcaaat attctggaa ggtacagat cttaatcgt gttacttta gccaccgct acttacacc cttcatccc aggtataat tccgagact ttggaaaat gatcttggt ttgattaaa ttgcgactt ccgattgag ttcaatcaa tatatcagt gttatcaca ctaccaagt agtttcacc cctgattat gtttccgtc actgcggtt ggatgggga caagtcgat gattttagt accgggctt tcgaacaat cttgtttcc gtcacactt actacaata tctcatgat gagtgtcag ttaacttac ggtccacaa attacgcat gatatgctc tgtgctgct ggaaacttc aatgagggt atttgtaat ggtgatatg ggtggagct ttaattcaa acaattggt ggaacagca gttcatgtt ggaattgca agtttcaga agtcacaac ggatgtgaa accactgat ccttcaggt tttactagg acaattgct tacgtagac tggataaga aattttgtt gcacttttt tccctattc acacttttt tcatcacca gtttatagt aacatagct ggacgcatt attggtggc agtgatgct aaagctgga gaattccca ttcactgca gcaatttac attcatacc agttcaggc cagtatttt tgtgctgga gctctgctt agccgcgag tggattttg accggtgct caatgtgtt gatggtgcc cttcagttt actatagtt cttggttct aacaatttg caagatagt aatgacccc aacagagtt aaagtagct actgaagag tactatctg tatccaaag tatgatgcg ttcactctt cagcatgat attgctctt attaagttg cgtctgcct atagaatat agcgattac attcaacca atcctttta ccaactgca gaactggtc gattattcg gtagttgta gcgcttggg tggggacaa accagtgat gaggatgcc ggcaatgtt tctgatctt cacagagtt agcatcacg gttttaagt aaccaggaa tgtcgaaac acttacggg actcaaatt atcgaagaa atggtctgt gctgaagga aattacaac caaggaact tgccatggt gatagtggt ggtcctttg attcaatat gtttacgga caatctgcc acacatgtc ggtgtgtct agctttttc agtggaaat gggtgtgaa tcaaccgat ccgtctggt tttacaagg acttttccc tatgttgaa tggataaga aatgtaaca caaatctaa
- >TC010933 RA
tttttttaa caaatatct agacaacca tattattga atttctgta ttcacaata aaagaatgt taagccgcc aaaccacta cttaatatt aaatcgttt tgacaatag aagataaat aatccagtt aagcatatc ctagattac ggtaaggat aaattacaa ttcacaaca tcaaagatc atttcatca tgaaattcc tcgcattct ttttgctgt gtgtaccac acgttttga gtttgccta aagggtctc gaattgttg gaggaaacc aggccagag gtggtcagt ttccctttg tggcggcaa tttatgtaa caaggtctg gaggtactt acttctgtg gaggtgttt tgcttagta atcagtggg ttctcacgg ctggacaat gcgtagatg gagccactt cgtttcgag ttcaacttg gctcaaata ttctggaag gtacagatc ttaatcgtg ttactttag ccaccgcta cttacaccc ttcatccca ggtataatt ccgagactt tggaaaatg atcttggtt tgattaaat tgcgacttc cgattgagt tcaatcaat atatcagtg ttatcacac taccaagta gtttcaccc ctgattatg tttccgtca ctgcggttg gatggggac aagtcgatg attttagta ccgggcttt cgaacaatc ttgtttccg tcacactta ctacaatat ctcatgatg agtgtcagt taacttacg gtccacaaa ttacgcatg atatgctct gtgctgctg gaaacttca atgagggta tttgtaatg gtgatatgg gtggagctt taattcaaa caattggtg gaacagcag ttcatgttg gaattgcaa gtttcagaa gtcacaacg gatgtgaaa ccactgatc cttcaggtt ttactagga caattgctt acgtagact ggataagaa attttgttg cactttttt ccctattca cactttttt catcaccag tttatagta acatagctg gacgcatta ttggtggca gtgatgcta aagctggag aattcccat tcactgcag caatttaca ttcatacca gttcaggcc agtattttt gtgctggag ctctgctta gccgcgagt ggattttga ccggtgctc aatgtgttg atggtgccc ttcagttta ctatagttc ttggttcta acaatttgc aagatagta atgacccca acagagtta aagtagcta ctgaagagt actatctgt atccaaagt atgatgcgt tcactcttc agcatgata ttgctctta ttaagttgc gtctgccta tagaatata gcgattaca ttcaaccaa tccttttac caactgcag aactggtcg attattcgg tagttgtag cgcttgggt ggggacaaa ccagtgatg aggatgccg gcaatgttt ctgatcttc acagagtta gcatcacgg ttttaagta accaggaat gtcgaaaca cttacggga ctcaaatta tcgaagaaa tggtctgtg ctgaaggaa attacaacc aaggaactt gccatggtg atagtggtg gtcctttga ttcaatatg tttacggac aatctgcca cacatgtcg gtgtgtcta gctttttca gtggaaatg ggtgtgaat caaccgatc cgtctggtt ttacaagga cttttccct atgttgaat ggataagaa atgtaacac aaatctaag aaataaatt tttggaaat tac
