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_02984 | 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: 30.0% (includes death as pupa, adult)
adult lethality - Number of animals showing the phenotype: 3 (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: % (includes death as larva, prepupa, pupa)
Lethalities 22 days after larval injection: % (includes death as larva, prepupa, pupa, adult)
- >TC000148 RA
MMFVIFFAV LFLIYYYLT RNFKHWEKK NVPFIKPLP FFGSIYDGV LMRHSIGEV FYDLYYKST KPFVGFFIL DKPCLLIRD PKLIKKILV NDFQYFYDR NAVNNKRDD PISTHILFI LKNPDWREL RTKMTPVFT SSKIKIMSE LIENASHEM TNYLNNHIK DYNSVEMRD VCLKFTVDV IGSTIFGVQ ANSFKDENS QFSSVAKRL IDWDDIVTA FRFRCYLLA PLFVNLFRM KLFPPDCVN FLKNTFLDI MDKRSVSNK SRNDLIDIL LQMKNDNRN FIEGDILVS QALMFFVAG FETTSSTMG FALYEFARN PDIQDKIRN EIKDISDKY GDIKYDSLK EMEYLDMCV KEVLRKYPV VPFLDRKCN TTYTIPDTN VTIDKDTPI FIPSLALHY DPQYFPNAD IFDPERFSS NNKTGIDSF AYLPFGEGP RNCIGARFG LLTAKLGLV HILKEFVVS CNEKSNEKI KFNPKGMVL SALNGINLK LSKVEKAT
- >TC000148 RB
MMFVIFFAV LFLIYYYLT RNFKHWEKK NVPFIKPLP FFGSIYDGV LMRHSIGEV FYDLYYKST KPFVGFFIL DKPCLLIRD PKLIKKILV NDFQYFYDR NAVNNKRDD PISTHILFI LKNPDWREL RTKMTPVFT SSKIKIMSE LIENASHEM TNYLNNHIK DYNSVEMRD VCLKFTVDV IGSTIFGVQ ANSFKDENS QFSSVAKRL IDWDDIVTA FRFRCYLLA PLFVNLFRM KLFPPDCVN FLKNTFLDI MDKRSVSNK SRNDLIDIL LQMKNDNRN FIEGDILVS QALMFFVAG FETTSSTMG FALYEFARN PDIQDKIRN EIKDISDKY GDIKYDSLK EMEYLDMCV KGITNYGNL FNFFKFVIL EVLRKYPVV PFLDRKCNT TYTIPDTNV TIDKDTPIF IPSLALHYD PQYFPNADI FDPERFSSN NKTGIDSFA YLPFGEGPR NCIGARFGL LTAKLGLVH ILKEFVVSC NEKSNEKIK FNPKGMVLS ALNGINLKL SKVEKAT
- >TC000148 RA
atgatgttt gtaattttc tttgctgtg ttattcctc atctattac tatttgacc cgaaatttc aaacactgg gaaaagaaa aatgtgcct ttcatcaag cctttgcca ttttttggt agtatttac gatggtgtt ttgatgcgt cactcgata ggagaagta ttttacgac ctttattac aaatcaacg aaacctttc gttggattt tttatcctg gataagcct tgtttacta atcagagat ccgaaattg attaagaag attctagta aacgatttt caatatttc tatgacaga aatgccgta aataataaa agggatgac ccaattagt acccacatt ttattcata ttgaaaaat cccgactgg agggaactt aggaccaaa atgacgcca gtttttacc tcttcaaaa ataaaaata atgtcggaa ttaatcgaa aatgcttcg catgaaatg accaactat ttaaacaat catatcaag gattacaat tcagttgaa atgagagat gtttgtttg aaattcacc gtggatgtg attggttca actattttt ggagtgcaa gctaattct ttcaaagat gaaaattcg caatttagt tcagtggcc aagagactg attgattgg gacgatatt gtcacagca tttcggttc aggtgttat cttttggcc cctttgttt gttaacttg tttcgaatg aaactcttt cctcctgat tgtgtaaat tttttgaag aacactttt ttggatatc atggataag agatcggtt tcaaacaag tccagaaat gatttaatt gatatttta ctacaaatg aagaatgat aatcgcaac ttcatagaa ggagatatt ttagtgtct caagcactc atgttcttt gttgctggc tttgagaca acgagctca acaatgggc tttgcattg tatgaattt gcacgtaat cctgacatt caggacaaa ataagaaac gaaataaaa gacatttcg gacaaatac ggcgatata aaatatgat tccttaaaa gaaatggaa tatttggac atgtgtgtt aaggaagtt ttacgaaaa tatccagtt gtgccattc cttgacaga aaatgtaac accacctac acgatacca gacaccaat gttacgatt gataaagac actcctatt ttcatccca agtttagct ttgcactat gatccccaa tattttccg aatgctgat atttttgat ccggagcgc ttttccagt aataacaaa actggaatc gacagtttt gcttactta ccatttggg gaaggacca cgtaattgt ataggtgca agatttggt ttgctaacc gcaaaatta ggtctagtg catattttg aaagagttt gttgtgagt tgtaatgag aagagtaac gaaaagatt aaatttaat cccaaaggg atggtcctg tcggcgtta aatggaatt aatttgaaa ttaagcaaa gttgaaaaa gcaacttaa
- >TC000148 RB
atgatgttt gtaattttc tttgctgtg ttattcctc atctattac tatttgacc cgaaatttc aaacactgg gaaaagaaa aatgtgcct ttcatcaag cctttgcca ttttttggt agtatttac gatggtgtt ttgatgcgt cactcgata ggagaagta ttttacgac ctttattac aaatcaacg aaacctttc gttggattt tttatcctg gataagcct tgtttacta atcagagat ccgaaattg attaagaag attctagta aacgatttt caatatttc tatgacaga aatgccgta aataataaa agggatgac ccaattagt acccacatt ttattcata ttgaaaaat cccgactgg agggaactt aggaccaaa atgacgcca gtttttacc tcttcaaaa ataaaaata atgtcggaa ttaatcgaa aatgcttcg catgaaatg accaactat ttaaacaat catatcaag gattacaat tcagttgaa atgagagat gtttgtttg aaattcacc gtggatgtg attggttca actattttt ggagtgcaa gctaattct ttcaaagat gaaaattcg caatttagt tcagtggcc aagagactg attgattgg gacgatatt gtcacagca tttcggttc aggtgttat cttttggcc cctttgttt gttaacttg tttcgaatg aaactcttt cctcctgat tgtgtaaat tttttgaag aacactttt ttggatatc atggataag agatcggtt tcaaacaag tccagaaat gatttaatt gatatttta ctacaaatg aagaatgat aatcgcaac ttcatagaa ggagatatt ttagtgtct caagcactc atgttcttt gttgctggc tttgagaca acgagctca acaatgggc tttgcattg tatgaattt gcacgtaat cctgacatt caggacaaa ataagaaac gaaataaaa gacatttcg gacaaatac ggcgatata aaatatgat tccttaaaa gaaatggaa tatttggac atgtgtgtt aagggtata acgaattat ggtaactta tttaatttt tttaaattc gtaatttta gaagtttta cgaaaatat ccagttgtg ccattcctt gacagaaaa tgtaacacc acctacacg ataccagac accaatgtt acgattgat aaagacact cctattttc atcccaagt ttagctttg cactatgat ccccaatat tttccgaat gctgatatt tttgatccg gagcgcttt tccagtaat aacaaaact ggaatcgac agttttgct tacttacca tttggggaa ggaccacgt aattgtata ggtgcaaga tttggtttg ctaaccgca aaattaggt ctagtgcat attttgaaa gagtttgtt gtgagttgt aatgagaag agtaacgaa aagattaaa tttaatccc aaagggatg gtcctgtcg gcgttaaat ggaattaat ttgaaatta agcaaagtt gaaaaagca acttaa
- >TC000148 RA
aaaattaaa attcctccc atccattca gatgtatct cctttggcc gtgaaccgt ttgcgtata ataaagtga aggtgacga tgatgtttg taattttct ttgctgtgt tattcctca tctattact atttgaccc gaaatttca aacactggg aaaagaaaa atgtgcctt tcatcaagc ctttgccat tttttggta gtatttacg atggtgttt tgatgcgtc actcgatag gagaagtat tttacgacc tttattaca aatcaacga aacctttcg ttggatttt ttatcctgg ataagcctt gtttactaa tcagagatc cgaaattga ttaagaaga ttctagtaa acgattttc aatatttct atgacagaa atgccgtaa ataataaaa gggatgacc caattagta cccacattt tattcatat tgaaaaatc ccgactgga gggaactta ggaccaaaa tgacgccag tttttacct cttcaaaaa taaaaataa tgtcggaat taatcgaaa atgcttcgc atgaaatga ccaactatt taaacaatc atatcaagg attacaatt cagttgaaa tgagagatg tttgtttga aattcaccg tggatgtga ttggttcaa ctatttttg gagtgcaag ctaattctt tcaaagatg aaaattcgc aatttagtt cagtggcca agagactga ttgattggg acgatattg tcacagcat ttcggttca ggtgttatc ttttggccc ctttgtttg ttaacttgt ttcgaatga aactctttc ctcctgatt gtgtaaatt ttttgaaga acacttttt tggatatca tggataaga gatcggttt caaacaagt ccagaaatg atttaattg atattttac tacaaatga agaatgata atcgcaact tcatagaag gagatattt tagtgtctc aagcactca tgttctttg ttgctggct ttgagacaa cgagctcaa caatgggct ttgcattgt atgaatttg cacgtaatc ctgacattc aggacaaaa taagaaacg aaataaaag acatttcgg acaaatacg gcgatataa aatatgatt ccttaaaag aaatggaat atttggaca tgtgtgtta aggaagttt tacgaaaat atccagttg tgccattcc ttgacagaa aatgtaaca ccacctaca cgataccag acaccaatg ttacgattg ataaagaca ctcctattt tcatcccaa gtttagctt tgcactatg atccccaat attttccga atgctgata tttttgatc cggagcgct tttccagta ataacaaaa ctggaatcg acagttttg cttacttac catttgggg aaggaccac gtaattgta taggtgcaa gatttggtt tgctaaccg caaaattag gtctagtgc atattttga aagagtttg ttgtgagtt gtaatgaga agagtaacg aaaagatta aatttaatc ccaaaggga tggtcctgt cggcgttaa atggaatta atttgaaat taagcaaag ttgaaaaag caacttaat aaaatattt ttggcacaa aatttcgtg tagttttag tacttg
- >TC000148 RB
aaaattaaa attcctccc atccattca gatgtatct cctttggcc gtgaaccgt ttgcgtata ataaagtga aggtgacga tgatgtttg taattttct ttgctgtgt tattcctca tctattact atttgaccc gaaatttca aacactggg aaaagaaaa atgtgcctt tcatcaagc ctttgccat tttttggta gtatttacg atggtgttt tgatgcgtc actcgatag gagaagtat tttacgacc tttattaca aatcaacga aacctttcg ttggatttt ttatcctgg ataagcctt gtttactaa tcagagatc cgaaattga ttaagaaga ttctagtaa acgattttc aatatttct atgacagaa atgccgtaa ataataaaa gggatgacc caattagta cccacattt tattcatat tgaaaaatc ccgactgga gggaactta ggaccaaaa tgacgccag tttttacct cttcaaaaa taaaaataa tgtcggaat taatcgaaa atgcttcgc atgaaatga ccaactatt taaacaatc atatcaagg attacaatt cagttgaaa tgagagatg tttgtttga aattcaccg tggatgtga ttggttcaa ctatttttg gagtgcaag ctaattctt tcaaagatg aaaattcgc aatttagtt cagtggcca agagactga ttgattggg acgatattg tcacagcat ttcggttca ggtgttatc ttttggccc ctttgtttg ttaacttgt ttcgaatga aactctttc ctcctgatt gtgtaaatt ttttgaaga acacttttt tggatatca tggataaga gatcggttt caaacaagt ccagaaatg atttaattg atattttac tacaaatga agaatgata atcgcaact tcatagaag gagatattt tagtgtctc aagcactca tgttctttg ttgctggct ttgagacaa cgagctcaa caatgggct ttgcattgt atgaatttg cacgtaatc ctgacattc aggacaaaa taagaaacg aaataaaag acatttcgg acaaatacg gcgatataa aatatgatt ccttaaaag aaatggaat atttggaca tgtgtgtta agggtataa cgaattatg gtaacttat ttaattttt ttaaattcg taattttag aagttttac gaaaatatc cagttgtgc cattccttg acagaaaat gtaacacca cctacacga taccagaca ccaatgtta cgattgata aagacactc ctattttca tcccaagtt tagctttgc actatgatc cccaatatt ttccgaatg ctgatattt ttgatccgg agcgctttt ccagtaata acaaaactg gaatcgaca gttttgctt acttaccat ttggggaag gaccacgta attgtatag gtgcaagat ttggtttgc taaccgcaa aattaggtc tagtgcata ttttgaaag agtttgttg tgagttgta atgagaaga gtaacgaaa agattaaat ttaatccca aagggatgg tcctgtcgg cgttaaatg gaattaatt tgaaattaa gcaaagttg aaaaagcaa cttaataaa atatttttg gcacaaaat ttcgtgtag ttttagtac ttg
