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_00436 | 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: 30.0% (includes death as larva, prepupa, pupa)
Lethalities 22 days after larval injection: 30.0% (includes death as larva, prepupa, pupa, adult)
- >TC034599 RA
MCLLLVIVT LVTLLLPLH FTTTPSPTT ATTTAVSNY PYVAFLQNS PTNAWCAGT LISNDTVLT AAHCIYGAN TLSVLLGAN DLTEDSNAV KLRSSEFAI HENYTNTTN SYMHDIGLI FLSQSIMLN DKIDTVPLS NSPFFYTTE KTCYVLGWK TSDNNFSET LLEKIVTLR EECDPFKIC AFGENGPFE GDSGGPLIC ESMQIGIVS GGTVEDNQL SIYVDVGSY NDWISEQIL QNSPTFKIS TLIVSSLLT KMSKEAILL IVLALSHYS VCGSNLELI QAHIEPSKV RIIGGDEVV PHSVPYQVG LRINGNAFC GGALISPNY VLTAGHCGE VIRSVEVIL GAHNISNPS EDTQVTITG SKITNHEKY DSATLRNDI CLIQLSEPA PINDNIQAA KLPPSSDSG KSYFDETVT ATGWGLYKD VPFPTTRDM SDVLLKVEV QVSNLTECG MFYNDDDGT YVVDTNLCT SGYRNKGTC NGDSGGPLS LDGVLIGLT SFGTDLCEM CSPSVYTRV VDYLDWIAQ NSDVHID
- >TC034599 RA
atgtgctta ttacttgtt attgttact cttgtcaca ttacttttg ccactacat tttactaca actccgagt ccaaccact gctaccacc actgcagtt tcgaactac ccatatgtg gccttttta cagaattct cccacaaac gcttggtgt gctggaact ttaatttcg aatgataca gttttaaca gcagctcat tgtatttac ggtgcaaac acgctttca gttctacta ggagctaat gatctaacc gaagattca aatgctgta aagttgaga agttccgaa tttgctatt cacgagaat tatactaat accacaaat tcttatatg catgatatt gggcttatt ttcttaagt cagtcaatt atgcttaat gacaaaatt gatactgtt ccgttgtct aatagtccg tttttctat acaacggag aaaacctgt tatgttttg ggttggaaa acatcagat aacaacttc agtgaaaca ttattagaa aaaatcgtt acactaagg gaagaatgc gatccattc aaaatttgt gcttttggt gaaaatggc ccttttgaa ggagattcg ggaggacct cttatctgc gaatccatg caaattggt atagtttca ggtggaact gtggaggac aatcaactg tcaatatac gtggacgtt ggaagttac aacgattgg atcagtgaa caaatttta caaaatagt cctacgttt aaaatctca accttgatt gtttcatca cttttgacc aaaatgtcc aaagaagca atcctgttg attgtcttg gctctaagc cactatagt gtctgtggt agcaacctg gagctgatt caagcccac attgaacca tcaaaagtc cgaattatt ggaggcgac gaagtagta ccccattcc gtaccttac caggttggt ttaagaatc aatggaaat gccttctgt ggcggagct ctcatttcg ccaaattac gttctcaca gctggtcat tgtggcgaa gtgattcga tcagttgag gtaattttg ggtgctcac aatatttcc aacccttca gaagacact caagttaca ataactggg agcaaaatc acaaaccac gaaaaatat gattctgca acgcttcgt aacgatatt tgtttgatt cagttgtca gaaccagct cctattaat gacaatatt caagcagca aagctacca ccaagtagt gattcgggt aaaagttat tttgatgaa actgtcaca gcaactgga tgggggttg tacaaggat gtacctttt cccaccact agagacatg tctgatgtt ctgttgaaa gttgaggtt caagtttca aatttaact gaatgtgga atgttttac aatgatgat gatggtact tatgtagtt gacacaaac ctttgtacc tcgggatat agaaataaa gggacttgt aatggagat tctggtggt cctttgagt ctagatggg gtacttatt ggacttacc tctttcgga acggatttg tgtgaaatg tgcagtcct tcggtatac acaagagtc gtggattac cttgattgg attgctcaa aattctgat gttcacatt gattga
- >TC034599 RA
tgttccaga tttaaccct gtacgctgt gctcaaata accatgtgc ttattactt gttattgtt actcttgtc acattactt ttgccacta cattttact acaactccg agtccaacc actgctacc accactgca gtttcgaac tacccatat gtggccttt ttacagaat tctcccaca aacgcttgg tgtgctgga actttaatt tcgaatgat acagtttta acagcagct cattgtatt tacggtgca aacacgctt tcagttcta ctaggagct aatgatcta accgaagat tcaaatgct gtaaagttg agaagttcc gaatttgct attcacgag aattatact aataccaca aattcttat atgcatgat attgggctt attttctta agtcagtca attatgctt aatgacaaa attgatact gttccgttg tctaatagt ccgtttttc tatacaacg gagaaaacc tgttatgtt ttgggttgg aaaacatca gataacaac ttcagtgaa acattatta gaaaaaatc gttacacta agggaagaa tgcgatcca ttcaaaatt tgtgctttt ggtgaaaat ggccctttt gaaggagat tcgggagga cctcttatc tgcgaatcc atgcaaatt ggtatagtt tcaggtgga actgtggag gacaatcaa ctgtcaata tacgtggac gttggaagt tacaacgat tggatcagt gaacaaatt ttacaaaat agtcctacg tttaaaatc tcaaccttg attgtttca tcacttttg accaaaatg tccaaagaa gcaatcctg ttgattgtc ttggctcta agccactat agtgtctgt ggtagcaac ctggagctg attcaagcc cacattgaa ccatcaaaa gtccgaatt attggaggc gacgaagta gtaccccat tccgtacct taccaggtt ggtttaaga atcaatgga aatgccttc tgtggcgga gctctcatt tcgccaaat tacgttctc acagctggt cattgtggc gaagtgatt cgatcagtt gaggtaatt ttgggtgct cacaatatt tccaaccct tcagaagac actcaagtt acaataact gggagcaaa atcacaaac cacgaaaaa tatgattct gcaacgctt cgtaacgat atttgtttg attcagttg tcagaacca gctcctatt aatgacaat attcaagca gcaaagcta ccaccaagt agtgattcg ggtaaaagt tattttgat gaaactgtc acagcaact ggatggggg ttgtacaag gatgtacct tttcccacc actagagac atgtctgat gttctgttg aaagttgag gttcaagtt tcaaattta actgaatgt ggaatgttt tacaatgat gatgatggt acttatgta gttgacaca aacctttgt acctcggga tatagaaat aaagggact tgtaatgga gattctggt ggtcctttg agtctagat ggggtactt attggactt acctctttc ggaacggat ttgtgtgaa atgtgcagt ccttcggta tacacaaga gtcgtggat taccttgat tggattgct caaaattct gatgttcac attgattga ttaacaata aaacaatga cactcaa
