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_00206 | 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. |
Get FastA Get primers |
||
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.
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: 11.11% (includes death as larva, prepupa, pupa)
Lethalities 22 days after larval injection: 11.11% (includes death as larva, prepupa, pupa, adult)
eclosion not fulfilled (with pupal and adult features) - Number of animals showing the phenotype: 1 (19 dpi)
- >TC030995 RA
MKIWLCAKT LRKVSQDCL RRVDAWETL LGGFITQLT AALVGVSES SKSRFVESV KPIVQRPYF DDVGPRNVT AVVGQSALL NCRVKHPGD RRVSWMRKR DLHILTSDI HTYTGDARF SVRHPEHSD DWDLRIAYV QKRDAGVYE CQVNTEPKI NLAITLNVE DAQATISGP PEVYVKKGS TISLTCSVN VHSTPPSSV LWYHGASVV DFDSPRGGI SLETEKTEA GTTSKLLVT KALLSDSGN YTCMPSNAS PASAVVHVL NGEHPAAMQ TSRAGVRRQ MALLATFSL LVVTSVR
- >TC030995 RB
MKIWLCAKT LRKVSQDCL RRVDAWETL LGGFITQLT AALVGVSES SKSRFVESV KPIVQRPYF DDVGPRNVT AVVGQSALL NCRVKHPGD RRVSWMRKR DLHILTSDI HTYTGDARF SVRHPEHSD DWDLRIAYV QKRDAGVYE CQVNTEPKI NLAITLNVE DASSLAATH PDPRSKDAQ ATISGPPEV YVKKGSTIS LTCSVNVHS TPPSSVLWY HGASVVDFD SPRGGISLE TEKTEAGTT SKLLVTKAL LSDSGNYTC MPSNASPAS AVVHVLNGE HPAAMQTSR AGVRRQMAL LATFSLLVV TSVR
- >TC030995 RC
MITYLTSFT AALVGVSES SKSRFVESV KPIVQRPYF DDVGPRNVT AVVGQSALL NCRVKHPGD RRVSWMRKR DLHILTSDI HTYTGDARF SVRHPEHSD DWDLRIAYV QKRDAGVYE CQVNTEPKI NLAITLNVE DASSLAATH PDPRSKDAQ ATISGPPEV YVKKGSTIS LTCSVNVHS TPPSSVLWY HGASVVDFD SPRGGISLE TEKTEAGTT SKLLVTKAL LSDSGNYTC MPSNASPAS AVVHVLNGE HPAAMQTSR AGVRRQMAL LATFSLLVV TSVR
- >TC030995 RA
atgaaaata tggttgtgt gcgaaaact ttgcgaaaa gtgtcacaa gattgttta cgaagggtt gacgcctgg gaaactttg cttggaggg ttcataact caacttaca gctgccctg gtgggagtc tcggaaagt tccaagtcc cggtttgtg gaatcggtg aaaccgatc gtgcaaagg ccctatttc gatgacgtc ggccctaga aacgtcacc gccgtcgtg ggccagtca gctttgctc aattgccga gtcaaacat ccaggcgac agaagggtc tcctggatg cgtaaacgc gacctccac atcctgacc tctgacatc cacacatac accggagac gctcgtttc agcgtccgc catccggag cacagcgac gactgggat ctgcgcatt gcttacgtg cagaaacgc gacgccggc gtctacgag tgtcaggtg aacaccgag cccaagatt aatctcgcc atcacgttg aacgtcgag gatgcccag gcgacgatt tccggtccg ccggaagtg tacgtgaag aagggcagc acgatatcg ctcacgtgt tccgtgaac gtccattcg acccctccc tcgtccgtg ctgtggtac catggtgct tccgtggtg gacttcgac tcgcctcgt ggcggcatc agcctcgaa acggaaaaa actgaggca ggcaccacc agcaaatta ctggtcacc aaagctcta ctttccgac tccggaaat tacacttgc atgccgtcc aacgccagt cccgcatcg gccgtcgtt cacgtcctc aatggggag cacccggca gcgatgcag acgagtcgg gcaggcgtg cgacgtcaa atggccctt ttggcgaca ttttctctg ctggtggtg acctcagta agatga
- >TC030995 RB
atgaaaata tggttgtgt gcgaaaact ttgcgaaaa gtgtcacaa gattgttta cgaagggtt gacgcctgg gaaactttg cttggaggg ttcataact caacttaca gctgccctg gtgggagtc tcggaaagt tccaagtcc cggtttgtg gaatcggtg aaaccgatc gtgcaaagg ccctatttc gatgacgtc ggccctaga aacgtcacc gccgtcgtg ggccagtca gctttgctc aattgccga gtcaaacat ccaggcgac agaagggtc tcctggatg cgtaaacgc gacctccac atcctgacc tctgacatc cacacatac accggagac gctcgtttc agcgtccgc catccggag cacagcgac gactgggat ctgcgcatt gcttacgtg cagaaacgc gacgccggc gtctacgag tgtcaggtg aacaccgag cccaagatt aatctcgcc atcacgttg aacgtcgag gacgcgtcc tctcttgca gcaacacat ccggaccct cgctccaaa gatgcccag gcgacgatt tccggtccg ccggaagtg tacgtgaag aagggcagc acgatatcg ctcacgtgt tccgtgaac gtccattcg acccctccc tcgtccgtg ctgtggtac catggtgct tccgtggtg gacttcgac tcgcctcgt ggcggcatc agcctcgaa acggaaaaa actgaggca ggcaccacc agcaaatta ctggtcacc aaagctcta ctttccgac tccggaaat tacacttgc atgccgtcc aacgccagt cccgcatcg gccgtcgtt cacgtcctc aatggggag cacccggca gcgatgcag acgagtcgg gcaggcgtg cgacgtcaa atggccctt ttggcgaca ttttctctg ctggtggtg acctcagta agatga
- >TC030995 RC
atgattaca tatttgact agttttaca gctgccctg gtgggagtc tcggaaagt tccaagtcc cggtttgtg gaatcggtg aaaccgatc gtgcaaagg ccctatttc gatgacgtc ggccctaga aacgtcacc gccgtcgtg ggccagtca gctttgctc aattgccga gtcaaacat ccaggcgac agaagggtc tcctggatg cgtaaacgc gacctccac atcctgacc tctgacatc cacacatac accggagac gctcgtttc agcgtccgc catccggag cacagcgac gactgggat ctgcgcatt gcttacgtg cagaaacgc gacgccggc gtctacgag tgtcaggtg aacaccgag cccaagatt aatctcgcc atcacgttg aacgtcgag gacgcgtcc tctcttgca gcaacacat ccggaccct cgctccaaa gatgcccag gcgacgatt tccggtccg ccggaagtg tacgtgaag aagggcagc acgatatcg ctcacgtgt tccgtgaac gtccattcg acccctccc tcgtccgtg ctgtggtac catggtgct tccgtggtg gacttcgac tcgcctcgt ggcggcatc agcctcgaa acggaaaaa actgaggca ggcaccacc agcaaatta ctggtcacc aaagctcta ctttccgac tccggaaat tacacttgc atgccgtcc aacgccagt cccgcatcg gccgtcgtt cacgtcctc aatggggag cacccggca gcgatgcag acgagtcgg gcaggcgtg cgacgtcaa atggccctt ttggcgaca ttttctctg ctggtggtg acctcagta agatga
- >TC030995 RA
atagataat ttacgacgc aatgaaaat atggttgtg tgcgaaaac tttgcgaaa agtgtcaca agattgttt acgaagggt tgacgcctg ggaaacttt gcttggagg gttcataac tcaacttac agctgccct ggtgggagt ctcggaaag ttccaagtc ccggtttgt ggaatcggt gaaaccgat cgtgcaaag gccctattt cgatgacgt cggccctag aaacgtcac cgccgtcgt gggccagtc agctttgct caattgccg agtcaaaca tccaggcga cagaagggt ctcctggat gcgtaaacg cgacctcca catcctgac ctctgacat ccacacata caccggaga cgctcgttt cagcgtccg ccatccgga gcacagcga cgactggga tctgcgcat tgcttacgt gcagaaacg cgacgccgg cgtctacga gtgtcaggt gaacaccga gcccaagat taatctcgc catcacgtt gaacgtcga ggatgccca ggcgacgat ttccggtcc gccggaagt gtacgtgaa gaagggcag cacgatatc gctcacgtg ttccgtgaa cgtccattc gacccctcc ctcgtccgt gctgtggta ccatggtgc ttccgtggt ggacttcga ctcgcctcg tggcggcat cagcctcga aacggaaaa aactgaggc aggcaccac cagcaaatt actggtcac caaagctct actttccga ctccggaaa ttacacttg catgccgtc caacgccag tcccgcatc ggccgtcgt tcacgtcct caatgggga gcacccggc agcgatgca gacgagtcg ggcaggcgt gcgacgtca aatggccct tttggcgac attttctct gctggtggt gacctcagt aagatgatt acgctgtcc ttaagaaat atatcaatc aacgacacg ctcataacg aactagtgc taaaatcgg tgtaaataa aacaatttt gtgaaaact caggtggtg atatcgacg gatctgatt cgatctttg gtgctaaag tggacataa aaataataa aaggttgtg acgtgacga gggatgaac gtccatttc ttgtatata gtgtgattc atttgatat taatagact caaaagcac tttagatta ttctagcat tattcattg taacaatta ttattacat gttgatatt atgtagatt tattgttat cattaccaa attttatcg gactgggct ttttatttt agtcatcca tggtttgtc tttgctgct ctctcggtg tgaaacttt caatacaac tggaaactt tgaacgatt tattcccgg ctgttttac gtaaaattt taaaacatg tccgacaag aatatattt cgcttagag caactgtgt agtttgatt tatgcaacg acaaacttg gttgaacca caaaatttc ggaaaatca aatcttggc aatattact tcagaaatt gttaactaa tctaacatt ttattcata tttttataa aaatcgttc tacagtgta acaaaaaga atctcaact tgtaattat catcttgta catacaaca ataaaacac aaaatagtt c
- >TC030995 RB
atagataat ttacgacgc aatgaaaat atggttgtg tgcgaaaac tttgcgaaa agtgtcaca agattgttt acgaagggt tgacgcctg ggaaacttt gcttggagg gttcataac tcaacttac agctgccct ggtgggagt ctcggaaag ttccaagtc ccggtttgt ggaatcggt gaaaccgat cgtgcaaag gccctattt cgatgacgt cggccctag aaacgtcac cgccgtcgt gggccagtc agctttgct caattgccg agtcaaaca tccaggcga cagaagggt ctcctggat gcgtaaacg cgacctcca catcctgac ctctgacat ccacacata caccggaga cgctcgttt cagcgtccg ccatccgga gcacagcga cgactggga tctgcgcat tgcttacgt gcagaaacg cgacgccgg cgtctacga gtgtcaggt gaacaccga gcccaagat taatctcgc catcacgtt gaacgtcga ggacgcgtc ctctcttgc agcaacaca tccggaccc tcgctccaa agatgccca ggcgacgat ttccggtcc gccggaagt gtacgtgaa gaagggcag cacgatatc gctcacgtg ttccgtgaa cgtccattc gacccctcc ctcgtccgt gctgtggta ccatggtgc ttccgtggt ggacttcga ctcgcctcg tggcggcat cagcctcga aacggaaaa aactgaggc aggcaccac cagcaaatt actggtcac caaagctct actttccga ctccggaaa ttacacttg catgccgtc caacgccag tcccgcatc ggccgtcgt tcacgtcct caatgggga gcacccggc agcgatgca gacgagtcg ggcaggcgt gcgacgtca aatggccct tttggcgac attttctct gctggtggt gacctcagt aagatgatt acgctgtcc ttaagaaat atatcaatc aacgacacg ctcataacg aactagtgc taaaatcgg tgtaaataa aacaatttt gtgaaaact caggtggtg atatcgacg gatctgatt cgatctttg gtgctaaag tggacataa aaataataa aaggttgtg acgtgacga gggatgaac gtccatttc ttgtatata gtgtgattc atttgatat taatagact caaaagcac tttagatta ttctagcat tattcattg taacaatta ttattacat gttgatatt atgtagatt tattgttat cattaccaa attttatcg gactgggct ttttatttt agtcatcca tggtttgtc tttgctgct ctctcggtg tgaaacttt caatacaac tggaaactt tgaacgatt tattcccgg ctgttttac gtaaaattt taaaacatg tccgacaag aatatattt cgcttagag caactgtgt agtttgatt tatgcaacg acaaacttg gttgaacca caaaatttc ggaaaatca aatcttggc aatattact tcagaaatt gttaactaa tctaacatt ttattcata tttttataa aaatcgttc tacagtgta acaaaaaga atctcaact tgtaattat catcttgta catacaaca ataaaacac aaaatagtt c
- >TC030995 RC
ctgaaagtc gaaattgaa acacaagcg ccggtccct catgattac atatttgac tagttttac agctgccct ggtgggagt ctcggaaag ttccaagtc ccggtttgt ggaatcggt gaaaccgat cgtgcaaag gccctattt cgatgacgt cggccctag aaacgtcac cgccgtcgt gggccagtc agctttgct caattgccg agtcaaaca tccaggcga cagaagggt ctcctggat gcgtaaacg cgacctcca catcctgac ctctgacat ccacacata caccggaga cgctcgttt cagcgtccg ccatccgga gcacagcga cgactggga tctgcgcat tgcttacgt gcagaaacg cgacgccgg cgtctacga gtgtcaggt gaacaccga gcccaagat taatctcgc catcacgtt gaacgtcga ggacgcgtc ctctcttgc agcaacaca tccggaccc tcgctccaa agatgccca ggcgacgat ttccggtcc gccggaagt gtacgtgaa gaagggcag cacgatatc gctcacgtg ttccgtgaa cgtccattc gacccctcc ctcgtccgt gctgtggta ccatggtgc ttccgtggt ggacttcga ctcgcctcg tggcggcat cagcctcga aacggaaaa aactgaggc aggcaccac cagcaaatt actggtcac caaagctct actttccga ctccggaaa ttacacttg catgccgtc caacgccag tcccgcatc ggccgtcgt tcacgtcct caatgggga gcacccggc agcgatgca gacgagtcg ggcaggcgt gcgacgtca aatggccct tttggcgac attttctct gctggtggt gacctcagt aagatgatt acgctgtcc ttaagaaat atatcaatc aacgacacg ctcataacg aactagtgc taaaatcgg tgtaaataa aacaatttt gtgaaaact caggtggtg atatcgacg gatctgatt cgatctttg gtgctaaag tggacataa aaataataa aaggttgtg acgtgacga gggatgaac gtccatttc ttgtatata gtgtgattc atttgatat taatagact caaaagcac tttagatta ttctagcat tattcattg taacaatta ttattacat gttgatatt atgtagatt tattgttat cattaccaa attttatcg gactgggct ttttatttt agtcatcca tggtttgtc tttgctgct ctctcggtg tgaaacttt caatacaac tggaaactt tgaacgatt tattcccgg ctgttttac gtaaaattt taaaacatg tccgacaag aatatattt cgcttagag caactgtgt agtttgatt tatgcaacg acaaacttg gttgaacca caaaatttc ggaaaatca aatcttggc aatattact tcagaaatt gttaactaa tctaacatt ttattcata tttttataa aaatcgttc tacagtgta acaaaaaga atctcaact tgtaattat catcttgta catacaaca ataaaacac aaaatagtt c
