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.
| General information | ||||
| iB_00108 | 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: 0.0% (includes death as pupa, adult)
embryo/egg condensed embryonic tissue, no cuticle, cns fluorescence - Penetrance (percentage of animals/eggs on slide): 30% - 50%
abdomen musculature partially not present - Penetrance (percentage of animals/eggs on slide): less than 30%
affected eggs/embryos/larvae total percentage of affected eggs/embryos/larvae on slide - Penetrance (percentage of animals/eggs on slide): more than 80%
remarks: muscle pattern potentially obscured by segmentation defect,
egg without cuticle embryo/egg without visible larval cuticle in cuticle preparation - Penetrance (percentage of animals/eggs on slide): 50% - 80%
affected eggs/embryos/larvae - Penetrance (percentage of animals/eggs on slide): 50% - 80%
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: 20.0% (includes death as larva, prepupa, pupa)
Lethalities 22 days after larval injection: 20.0% (includes death as larva, prepupa, pupa, adult)
remarks: later than schedule, hours 100
- >TC000495 RA
MRCYRIQLF VVCSLLFFI NINAQRRSS CTTPNGDIA RCIPISSCP ILYDAVTTR DKQQLKFLK ESQCGYGRD PLVCCGLHN NYRLNPGDF AYDDEPFGW GNSHHRRNS GSQNWDRDD TPSWGNERK NNNNNNEWS TNNRRRTTT QRTNSEESG WGISDRPNR RTSGEESRW STSDRPNRR TSGEESGWG TTDRPNRRT SGEDSSRRN NNRRTTTQR TNSEESSFE REDDRFNDR TNSGSNSNR QTSQGSGST DKSECGVQE VDRILDGQA TDLREFPWM ALLQYRKKS GNLVFSCGG TLISPRYVL TAAHCVRGQ ILTKIGPLV NVRLGEYNT ETERDCSNQ MGFEICNEK PIDSEIDKV IPHPDYSDN SADRYHDIA LIKLKRQVS YTDFIKPIC LPGKSEKTS VGKRLAVAG WGRTEYASN SPVKLKLWV PVAETSQCS SKFKSAGVT LGNRQLCAG GEQGRDSCN GDSGGPLMA VRNATAQWY IEGIVSFGA RCGSEGWPG IYTRVSEYL DWIQNNTN
- >TC000495 RB
MRCYRIQLF VVCSLLFFI NINAQFFLE RRSSCTTPN GDIARCIPI SSCPILYDA VTTRDKQQL KFLKESQCG YGRDPLVCC GLHNNYRLN PGDFAYDDE PFGWGNSHH RRNSGSQNW DRDDTPSWG NERKNNNNN NEWSTNNRR RTTTQRTNS EESGWGISD RPNRRTSGE ESRWSTSDR PNRRTSGEE SGWGTTDRP NRRTSGEDS SRRNNNRRT TTQRTNSEE SSFEREDDR FNDRTNSGS NSNRQTSQG SGSTDKSEC GVQEVDRIL DGQATDLRE FPWMALLQY RKKSGNLVF SCGGTLISP RYVLTAAHC VRGQILTKI GPLVNVRLG EYNTETERD CSNQMGFEI CNEKPIDSE IDKVIPHPD YSDNSADRY HDIALIKLK RQVSYTDFI KPICLPGKS EKTSVGKRL AVAGWGRTE YASNSPVKL KLWVPVAET SQCSSKFKS AGVTLGNRQ LCAGGEQGR DSCNGDSGG PLMAVRNAT AQWYIEGIV SFGARCGSE GWPGIYTRV SEYLDWIQN NTN
- >TC000495 RA
atgcggtgt tacagaata cagttgttt gtggtttgt tcacttctt ttcttcatt aatattaac gcacaacga agatcgtca tgtacgact cctaacggc gacatagca cggtgtatc ccaattagc tcctgtcct attctttac gatgctgtc acaactcga gataagcaa cagttaaag tttttgaaa gaatcacag tgtggatat ggccgggat cctttggtt tgttgcggt cttcataac aattatcgt ctaaaccct ggcgatttt gcctacgat gatgaaccc ttcggctgg ggtaactcc caccataga aggaactca gggtcgcaa aactgggac agggacgat actccaagc tggggcaat gaaagaaaa aataataat aataataac gaatggagc actaataac aggagaaga actacaact caaagaaca aattcggaa gaatctgga tggggtatt agtgacaga ccaaacaga agaacaagc ggtgaagaa tctagatgg agtactagt gatagacca aacagaaga acaagcggt gaagaatct ggatggggt actactgac agaccaaat agacgaaca agcggtgaa gattcgagc cgtcgtaac aataacaga agaacgaca acacaacga acaaattcc gaggaatct agttttgaa agagaggat gacagattc aatgatcga actaattcg ggatcaaac agtaataga caaacaagt caaggttct ggtagtacc gataagtcc gaatgtgga gtccaggaa gttgatcga attctcgac gggcaggcc accgatttg cgagaattc ccttggatg gctttgtta cagtaccgc aaaaagtcc ggaaatttg gtattttcc tgtggcgga actctcatt tcacctcgc tacgtttta acagcagct cactgcgtc cgaggacaa atccttaca aaaatcggg ccattggtc aatgtcagg ctaggcgag tacaacact gaaacagag cgagactgt agtaaccaa atgggattc gaaatttgt aacgagaaa ccaatagat tcagagatt gataaagtg ataccccat cccgactac agcgataac agtgcggac agataccat gacattgct ttaatcaaa ttgaagcga caagtctcg tacactgat tttattaag cccatttgt ttgcccgga aaatcggaa aaaacgagt gttggtaaa cggctagcg gtagctgga tggggccga acggagtac gccagtaat agtccagtc aaactaaaa ctgtgggtt ccagtggcg gaaacctcc cagtgttcc agcaaattt aaatcggct ggagtgact ttagggaat agacagttg tgtgctgga ggagagcaa ggcagagat tcttgtaat ggggacagt ggtgggcct ttaatggcg gtcagaaat gcgacagcc caatggtac attgaagga attgtctcg tttggagcg agatgtggt agcgaagga tggccagga atttacaca cgggtgtcg gagtatctc gactggatt caaaacaat accaattaa
- >TC000495 RB
atgcggtgt tacagaata cagttgttt gtggtttgt tcacttctt ttcttcatt aatattaac gcacaattt tttttagaa cgaagatcg tcatgtacg actcctaac ggcgacata gcacggtgt atcccaatt agctcctgt cctattctt tacgatgct gtcacaact cgagataag caacagtta aagtttttg aaagaatca cagtgtgga tatggccgg gatcctttg gtttgttgc ggtcttcat aacaattat cgtctaaac cctggcgat tttgcctac gatgatgaa cccttcggc tggggtaac tcccaccat agaaggaac tcagggtcg caaaactgg gacagggac gatactcca agctggggc aatgaaaga aaaaataat aataataat aacgaatgg agcactaat aacaggaga agaactaca actcaaaga acaaattcg gaagaatct ggatggggt attagtgac agaccaaac agaagaaca agcggtgaa gaatctaga tggagtact agtgataga ccaaacaga agaacaagc ggtgaagaa tctggatgg ggtactact gacagacca aatagacga acaagcggt gaagattcg agccgtcgt aacaataac agaagaacg acaacacaa cgaacaaat tccgaggaa tctagtttt gaaagagag gatgacaga ttcaatgat cgaactaat tcgggatca aacagtaat agacaaaca agtcaaggt tctggtagt accgataag tccgaatgt ggagtccag gaagttgat cgaattctc gacgggcag gccaccgat ttgcgagaa ttcccttgg atggctttg ttacagtac cgcaaaaag tccggaaat ttggtattt tcctgtggc ggaactctc atttcacct cgctacgtt ttaacagca gctcactgc gtccgagga caaatcctt acaaaaatc gggccattg gtcaatgtc aggctaggc gagtacaac actgaaaca gagcgagac tgtagtaac caaatggga ttcgaaatt tgtaacgag aaaccaata gattcagag attgataaa gtgataccc catcccgac tacagcgat aacagtgcg gacagatac catgacatt gctttaatc aaattgaag cgacaagtc tcgtacact gattttatt aagcccatt tgtttgccc ggaaaatcg gaaaaaacg agtgttggt aaacggcta gcggtagct ggatggggc cgaacggag tacgccagt aatagtcca gtcaaacta aaactgtgg gttccagtg gcggaaacc tcccagtgt tccagcaaa tttaaatcg gctggagtg actttaggg aatagacag ttgtgtgct ggaggagag caaggcaga gattcttgt aatggggac agtggtggg cctttaatg gcggtcaga aatgcgaca gcccaatgg tacattgaa ggaattgtc tcgtttgga gcgagatgt ggtagcgaa ggatggcca ggaatttac acacgggtg tcggagtat ctcgactgg attcaaaac aataccaat taa
- >TC000495 RA
taaactcga aagaccagt ggcttgctt cttagttca acaaacatc gtcaaggcg gacggcagc gataaaatt atgcggtgt tacagaata cagttgttt gtggtttgt tcacttctt ttcttcatt aatattaac gcacaacga agatcgtca tgtacgact cctaacggc gacatagca cggtgtatc ccaattagc tcctgtcct attctttac gatgctgtc acaactcga gataagcaa cagttaaag tttttgaaa gaatcacag tgtggatat ggccgggat cctttggtt tgttgcggt cttcataac aattatcgt ctaaaccct ggcgatttt gcctacgat gatgaaccc ttcggctgg ggtaactcc caccataga aggaactca gggtcgcaa aactgggac agggacgat actccaagc tggggcaat gaaagaaaa aataataat aataataac gaatggagc actaataac aggagaaga actacaact caaagaaca aattcggaa gaatctgga tggggtatt agtgacaga ccaaacaga agaacaagc ggtgaagaa tctagatgg agtactagt gatagacca aacagaaga acaagcggt gaagaatct ggatggggt actactgac agaccaaat agacgaaca agcggtgaa gattcgagc cgtcgtaac aataacaga agaacgaca acacaacga acaaattcc gaggaatct agttttgaa agagaggat gacagattc aatgatcga actaattcg ggatcaaac agtaataga caaacaagt caaggttct ggtagtacc gataagtcc gaatgtgga gtccaggaa gttgatcga attctcgac gggcaggcc accgatttg cgagaattc ccttggatg gctttgtta cagtaccgc aaaaagtcc ggaaatttg gtattttcc tgtggcgga actctcatt tcacctcgc tacgtttta acagcagct cactgcgtc cgaggacaa atccttaca aaaatcggg ccattggtc aatgtcagg ctaggcgag tacaacact gaaacagag cgagactgt agtaaccaa atgggattc gaaatttgt aacgagaaa ccaatagat tcagagatt gataaagtg ataccccat cccgactac agcgataac agtgcggac agataccat gacattgct ttaatcaaa ttgaagcga caagtctcg tacactgat tttattaag cccatttgt ttgcccgga aaatcggaa aaaacgagt gttggtaaa cggctagcg gtagctgga tggggccga acggagtac gccagtaat agtccagtc aaactaaaa ctgtgggtt ccagtggcg gaaacctcc cagtgttcc agcaaattt aaatcggct ggagtgact ttagggaat agacagttg tgtgctgga ggagagcaa ggcagagat tcttgtaat ggggacagt ggtgggcct ttaatggcg gtcagaaat gcgacagcc caatggtac attgaagga attgtctcg tttggagcg agatgtggt agcgaagga tggccagga atttacaca cgggtgtcg gagtatctc gactggatt caaaacaat accaattaa actgactct catagttgt tgtgtatat ttatttatg atagcgctt aattaagtt cggaataaa acattaaat taacggacc agaaatttt ctcaagtga ggtacaagt gtattttat attaaagtt tatcaactg gtgataacc ttataaaaa tacttaaca gcatcgtag ttaatgcca tcgtcaata agcttgtta tgaaaacaa gagacgtat ttaattaaa aaaataata aaattttag tgaaatt
- >TC000495 RB
taaactcga aagaccagt ggcttgctt cttagttca acaaacatc gtcaaggcg gacggcagc gataaaatt atgcggtgt tacagaata cagttgttt gtggtttgt tcacttctt ttcttcatt aatattaac gcacaattt tttttagaa cgaagatcg tcatgtacg actcctaac ggcgacata gcacggtgt atcccaatt agctcctgt cctattctt tacgatgct gtcacaact cgagataag caacagtta aagtttttg aaagaatca cagtgtgga tatggccgg gatcctttg gtttgttgc ggtcttcat aacaattat cgtctaaac cctggcgat tttgcctac gatgatgaa cccttcggc tggggtaac tcccaccat agaaggaac tcagggtcg caaaactgg gacagggac gatactcca agctggggc aatgaaaga aaaaataat aataataat aacgaatgg agcactaat aacaggaga agaactaca actcaaaga acaaattcg gaagaatct ggatggggt attagtgac agaccaaac agaagaaca agcggtgaa gaatctaga tggagtact agtgataga ccaaacaga agaacaagc ggtgaagaa tctggatgg ggtactact gacagacca aatagacga acaagcggt gaagattcg agccgtcgt aacaataac agaagaacg acaacacaa cgaacaaat tccgaggaa tctagtttt gaaagagag gatgacaga ttcaatgat cgaactaat tcgggatca aacagtaat agacaaaca agtcaaggt tctggtagt accgataag tccgaatgt ggagtccag gaagttgat cgaattctc gacgggcag gccaccgat ttgcgagaa ttcccttgg atggctttg ttacagtac cgcaaaaag tccggaaat ttggtattt tcctgtggc ggaactctc atttcacct cgctacgtt ttaacagca gctcactgc gtccgagga caaatcctt acaaaaatc gggccattg gtcaatgtc aggctaggc gagtacaac actgaaaca gagcgagac tgtagtaac caaatggga ttcgaaatt tgtaacgag aaaccaata gattcagag attgataaa gtgataccc catcccgac tacagcgat aacagtgcg gacagatac catgacatt gctttaatc aaattgaag cgacaagtc tcgtacact gattttatt aagcccatt tgtttgccc ggaaaatcg gaaaaaacg agtgttggt aaacggcta gcggtagct ggatggggc cgaacggag tacgccagt aatagtcca gtcaaacta aaactgtgg gttccagtg gcggaaacc tcccagtgt tccagcaaa tttaaatcg gctggagtg actttaggg aatagacag ttgtgtgct ggaggagag caaggcaga gattcttgt aatggggac agtggtggg cctttaatg gcggtcaga aatgcgaca gcccaatgg tacattgaa ggaattgtc tcgtttgga gcgagatgt ggtagcgaa ggatggcca ggaatttac acacgggtg tcggagtat ctcgactgg attcaaaac aataccaat taaactgac tctcatagt tgttgtgta tatttattt atgatagcg cttaattaa gttcggaat aaaacatta aattaacgg accagaaat tttctcaag tgaggtaca agtgtattt tatattaaa gtttatcaa ctggtgata accttataa aaatactta acagcatcg tagttaatg ccatcgtca ataagcttg ttatgaaaa caagagacg tatttaatt aaaaaaata ataaaattt tagtgaaat t
