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_05412 | 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: 40.0% (includes death as pupa, adult)
pupa eclosion not fulfilled (died as pupa) - Number of animals showing the phenotype: 1 (3 dpi)eclosion not fulfilled (with pupal and adult features) - Number of animals showing the phenotype: 1 (3 dpi)
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: 20.0% (includes death as larva, prepupa, pupa)
Lethalities 22 days after larval injection: 20.0% (includes death as larva, prepupa, pupa, adult)
- >TC013277 RA
MLVRSLFIL VVTAQVLNA DENCRTPDN EEGDCKPIN KCQPLYSLL ERRPITAST ADYLRRSQC GFVGTYPKV CCPSGRTTI TTNPPPVVE GPTENTDVE SVTSNLLPG GDVCGLNTQ SRIYGGEKT DLDEFPWMA LIEYEKPGG SRGFYCGGV LISNKYILT AAHCVKGKD LPKTWKLVS VRLGEYNTE TDQDCINNG FGEDCAPPP VNVPVVERI AHESYDPND VNQYHDIAL LRLKRSVTF SDYVRPICL PTSNEELRR SFIGQKLFV AGWGKTENR SESNIKLKV QVPVKQTSE CSSTYRVAN VRLGPGQMC AGGEKGRDS CRGDSGGPL MTVIRDKNK DDHWYAAGV VSFGPSPCG MENWPGVYT KVSKYVNWI VNKLKP
- >TC013277 RA
atgttagtg aggagtctc tttattctg gtagtgaca gcacaagtt ttaaacgca gatgagaat tgccgaaca ccggataat gaggagggc gactgcaag cccatcaac aaatgccaa cccctgtac tccctctta gaaaggcgt cccatcaca gccagcact gccgattac cttcgccga tcgcaatgt ggtttcgtc ggcacttac cccaaagtg tgttgtcct tctggacgg acaactatt accacaaat ccgccaccg gttgtagaa ggaccgact gaaaacacc gatgttgaa tcggtgact agcaatttg ctccctggg ggcgacgtt tgcggccta aacacacaa agcaggatc tacggaggg gagaaaaca gacttggac gaattccct tggatggct ctgattgaa tacgaaaaa cctggtggt agccgaggt ttctactgc ggtggggtt ttaatcagt aacaaatac attttaacg gcggctcat tgcgtcaaa ggcaaggac ttgccaaag acgtggaaa ctagtgagc gtccgcttg ggcgaatac aacactgaa actgaccaa gactgcatc aacaacggc tttggtgaa gactgtgct ccaccccca gtcaacgtc cctgtagtt gaaagaata gcacatgaa agttacgat ccgaatgac gtaaaccag tatcacgat atcgccttg ctgaggctg aaacgctct gtgacattt tcagactat gtccgaccc atctgctta ccaacctca aacgaagaa cttcgacgg tcattcatt ggccaaaaa ctgtttgtc gccggttgg ggcaaaact gagaacagg tctgaaagc aacataaaa ctgaaagtc caggttcca gttaaacaa acttccgag tgtagctct acatatcgc gtcgctaat gtgagactt ggcccagga cagatgtgt gcaggtggt gaaaaaggg cgcgattct tgtagaggt gatagtggt ggtccgctg atgaccgtt atcagagac aaaaacaag gacgaccat tggtacgct gcaggagtc gtttcattt ggaccgtcg ccgtgtgga atggaaaat tggccagga gtttatacc aaagtttct aaatacgtc aactggatt gttaataag ctaaaacct taa
- >TC013277 RA
caacaacaa atgaccaat atttactcc attagcaat tataaacag aaagagatg acaaaatgt gtaaaattt gcaaagagg ggcgatgga gttttgatg acgtcttcc aagccatta gtatttcca gttcataga acagacggt ggactttcc aaggcaggc gagatactc acacaagat cgctcgaca tcggtatgg gtcagtgag aacccaagg cactcatat tattcataa ttgcaactt tgtttacaa tgttagtga ggagtctct ttattctgg tagtgacag cacaagttt taaacgcag atgagaatt gccgaacac cggataatg aggagggcg actgcaagc ccatcaaca aatgccaac ccctgtact ccctcttag aaaggcgtc ccatcacag ccagcactg ccgattacc ttcgccgat cgcaatgtg gtttcgtcg gcacttacc ccaaagtgt gttgtcctt ctggacgga caactatta ccacaaatc cgccaccgg ttgtagaag gaccgactg aaaacaccg atgttgaat cggtgacta gcaatttgc tccctgggg gcgacgttt gcggcctaa acacacaaa gcaggatct acggagggg agaaaacag acttggacg aattccctt ggatggctc tgattgaat acgaaaaac ctggtggta gccgaggtt tctactgcg gtggggttt taatcagta acaaataca ttttaacgg cggctcatt gcgtcaaag gcaaggact tgccaaaga cgtggaaac tagtgagcg tccgcttgg gcgaataca acactgaaa ctgaccaag actgcatca acaacggct ttggtgaag actgtgctc cacccccag tcaacgtcc ctgtagttg aaagaatag cacatgaaa gttacgatc cgaatgacg taaaccagt atcacgata tcgccttgc tgaggctga aacgctctg tgacatttt cagactatg tccgaccca tctgcttac caacctcaa acgaagaac ttcgacggt cattcattg gccaaaaac tgtttgtcg ccggttggg gcaaaactg agaacaggt ctgaaagca acataaaac tgaaagtcc aggttccag ttaaacaaa cttccgagt gtagctcta catatcgcg tcgctaatg tgagacttg gcccaggac agatgtgtg caggtggtg aaaaagggc gcgattctt gtagaggtg atagtggtg gtccgctga tgaccgtta tcagagaca aaaacaagg acgaccatt ggtacgctg caggagtcg tttcatttg gaccgtcgc cgtgtggaa tggaaaatt ggccaggag tttatacca aagtttcta aatacgtca actggattg ttaataagc taaaacctt aatttagag gattgttgt aaatgtgat ttttattaa gaaaatcgg agctaataa agttttttg aactat
