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_08283 | 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)
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)
- >TC004160 RA
MTTKPILNC FILLIVCCS SAFAQVGDT CVVKSTGVR GVCRISSNC PIVEEQAQN GISPTICGY YQLTVPIVC CEESPFVQP NDDTLLNPE PNLSNQQPS YSNSNSREP SYSNSNSRE PSYSNSNSR EPSYSNSNS REPSYSNSN SREPSYSNS NSREPSYSN SNSREPSYS NSNNRQPSP SQNREPNPN IFPAGEPDA VIYPDSDNQ PSRPQVTHT SKSEQKCQE YSKAITGVV QAIPLVTNT EVVSYSFVK CDYNGVALI VGGKPASAG EFPFMAAIG FYVDNKVEW RCGGTLISE EYVLTAAHC TYTRDGDTP KIVRLGDLD LSRDDDGSV HTDYNVRNI VVHPRYRYP LKYNDIALI QLSTTVRFT KFIRPACLY TKSQVELPQ AIATGWGKT DYAAAEISD KLMKVSLNI YSNDRCAQT YQTSKHLPQ GIKSNMICA GELRGGQDT CQGDSGGPL LITKKGNQC KFYVIGVTS FGKSCGQAN TPAIYTRVS EYVPWIEKT IW
- >TC004160 RA
atgacaaca aaacctata cttaactgt tttatattg ttaattgtt tgttgctct tctgcgttt gcgcaagtg ggagatacc tgcgtagta aaatccaca ggcgttcga ggggtttgc aggatatcc tcaaactgc cccattgtc gaagaacaa gcacaaaac ggaattagc ccgacaata tgtgggtac tatcaactg acagtcccc atcgtctgc tgcgaggaa agccccttc gtccaacca aacgacgat acgttactg aatccagaa cccaacctg agtaaccag caaccgagc tactccaac tccaacagc cgggaacca agctactca aactccaac agccgggaa ccgagctac tctaattcc aacagccgg gaaccgagc tactccaat tccaacagc cgggaaccg agctactcc aattccaac agccgggaa ccgagctac tccaattcc aacagccgg gaaccgagc tactccaat tccaacagc cgggaaccg agctactcc aactccaac aaccggcag ccaagtccc tcccaaaac cgggaaccg aatccgaac atatttcca gctggtgag ccggacgct gtgatatat cctgattcg gacaatcag ccgtcgcgg ccccaagtg actcataca agtaaaagc gagcaaaag tgccaagag tacagtaaa gccataact ggtgtcgtt caagcgatt cctttagtg actaatact gaggtggtt agctatagc tttgtcaag tgtgactac aatggtgta gcgttgatc gtcgggggc aagcctgcc tccgctggg gaattcccg tttatggcg gccatcggt ttttacgtg gataacaaa gtggagtgg cgctgcggg ggcacgctg atcagtgag gagtatgtt ttaaccgcc gcccactgc acttacacc cgcgatggg gacacgccg aagattgtt cgattggga gaccttgac ttatcccgc gatgacgac ggctccgta cacactgac tacaacgta cggaacatt gtagtgcat cctcggtac cgctacccc ctcaaatac aacgacatt gccctcatc cagctgtcc acaacagtc agattcacg aaattcatc cgacctgcg tgtctttac accaaatcg caggtggag ctcccgcaa gccatagcg accggctgg ggcaaaacg gactacgcc gctgccgag ataagcgac aagttgatg aaagtctcg ctaaatatt tacagcaat gaccgatgt gcccagacg taccagacc agcaaacac ttgccacaa gggatcaag tctaacatg atctgtgca ggggagttg agggggggc caggacacg tgtcagggg gactctggg gggccccta cttatcacc aaaaagggc aaccaatgc aagttctac gttatcggg gtcacctcg ttcgggaag tcctgcgga caggccaac acgcccgcc atctacacc agagtctca gaatacgtg ccatggatc gaaaaaacc atctggtaa
- >TC004160 RA
gtagaaagt ctaaagttt ccgtttcac gaattgtgt gaggtgttt gtgagacag cgatttagt cggtttttc ggtgaaaga gacgtttga tgaattttc aagatcttg ttaagaatc acatcagct tataaatag taataaaac atattgttt tcagttatt aatgagaca gtggtgtag tcaagttag cggaataat tattgtaat ttaacgaat ttatataag tttcttaat attgaggat cttcaagga ttttgttat ttatgcgag cgtgtcacg gtaaaccag atagcctgt ggtaaacaa taattgaag agtatattt tatgaaaca agttatagt gacgccaat gactgctgg taaatccag tgttattta catacatct taactaaaa agaaagaaa aagacgcag aatagtttc tgaacctga ggaataacg atgtcagtg ttttgaagt tcttcacaa tgacaacaa aacctatac ttaactgtt ttatattgt taattgttt gttgctctt ctgcgtttg cgcaagtgg gagatacct gcgtagtaa aatccacag gcgttcgag gggtttgca ggatatcct caaactgcc ccattgtcg aagaacaag cacaaaacg gaattagcc cgacaatat gtgggtact atcaactga cagtcccca tcgtctgct gcgaggaaa gccccttcg tccaaccaa acgacgata cgttactga atccagaac ccaacctga gtaaccagc aaccgagct actccaact ccaacagcc gggaaccaa gctactcaa actccaaca gccgggaac cgagctact ctaattcca acagccggg aaccgagct actccaatt ccaacagcc gggaaccga gctactcca attccaaca gccgggaac cgagctact ccaattcca acagccggg aaccgagct actccaatt ccaacagcc gggaaccga gctactcca actccaaca accggcagc caagtccct cccaaaacc gggaaccga atccgaaca tatttccag ctggtgagc cggacgctg tgatatatc ctgattcgg acaatcagc cgtcgcggc cccaagtga ctcatacaa gtaaaagcg agcaaaagt gccaagagt acagtaaag ccataactg gtgtcgttc aagcgattc ctttagtga ctaatactg aggtggtta gctatagct ttgtcaagt gtgactaca atggtgtag cgttgatcg tcgggggca agcctgcct ccgctgggg aattcccgt ttatggcgg ccatcggtt tttacgtgg ataacaaag tggagtggc gctgcgggg gcacgctga tcagtgagg agtatgttt taaccgccg cccactgca cttacaccc gcgatgggg acacgccga agattgttc gattgggag accttgact tatcccgcg atgacgacg gctccgtac acactgact acaacgtac ggaacattg tagtgcatc ctcggtacc gctaccccc tcaaataca acgacattg ccctcatcc agctgtcca caacagtca gattcacga aattcatcc gacctgcgt gtctttaca ccaaatcgc aggtggagc tcccgcaag ccatagcga ccggctggg gcaaaacgg actacgccg ctgccgaga taagcgaca agttgatga aagtctcgc taaatattt acagcaatg accgatgtg cccagacgt accagacca gcaaacact tgccacaag ggatcaagt ctaacatga tctgtgcag gggagttga gggggggcc aggacacgt gtcaggggg actctgggg ggcccctac ttatcacca aaaagggca accaatgca agttctacg ttatcgggg tcacctcgt tcgggaagt cctgcggac aggccaaca cgcccgcca tctacacca gagtctcag aatacgtgc catggatcg aaaaaacca tctggtaaa atcacccac ttaacgaca ttttttatt ttatcattc acatacaat acgtataga
