Biology subjects

King-Jones, K.

Publications and source records attributed to King-Jones, K..

4 recordsLinked to original sources

bioRxiv2025

Molecular Arms Race: Tannin Biosynthesis and Laccase-Based Detoxification in the Aphid-Gall System

O_LIPlant-insect coevolution is exemplified by Schlechtendalia chinensis inducing gallnuts with record-breaking hydrolyzable tannin (HT) concentrations of 74.49%--32-fold higher than normal leaves. How do plants achieve this extreme defensive chemistry, and how do aphids survive it? C_LIO_LIWe integrated transcriptomics, heterologous gene validation in Arabidopsis, and enzyme assays to investigate both plant HT biosynthesis and aphid detoxification mechanisms throughout gall development. C_LIO_LIThree key genes--Phosphoglucomutase (PGM), UDP-glucosyltransferase BX9 (BX9), and gallate 1-beta-D-glucosyltransferase (GDG) -- govern HT biosynthesis, with expression patterns closely tracking tannin accumulation dynamics, and GDG as the rate-limiting enzyme. Arabidopsis transformants showed threefold HT increases. Critically, S. chinensis employs specialized laccases rather than tannase for detoxification, with laccase activity exceeding tannase by 20,000-fold. The aphid genome encodes three laccase genes, with Sc-Lac1 expressed in digestive tissues, achieving 39-54% HT degradation. C_LIO_LIThese findings reveal how plants weaponize secondary metabolism while herbivores evolve enzymatic countermeasures. The identified genes enable engineering enhanced defenses or pharmaceutical tannin production, while laccase-based detoxification offers new insights into insect adaptation to chemical defenses. C_LI

evolutionary biology↗

bioRxiv2020

A versatile toolkit for CRISPR-Cas13-based RNA manipulation in Drosophila

Advances in CRISPR technology have immensely improved our ability to manipulate nucleic acids, and the recent discovery of the RNA-targeting endonuclease Cas13 adds even further functionality. Here, we show that Cas13 works efficiently in Drosophila, both ex vivo and in vivo. We tested 44 different Cas13 variants to identify enzymes with the best overall performance and showed that Cas13 could target endogenous Drosophila transcripts in vivo with high efficiency and specificity. We also developed Cas13 applications to edit mRNAs and target mitochondrial transcripts. Our vector collection represents a versatile tool collection to manipulate gene expression at the post-transcriptional level.

molecular biology↗

bioRxiv2018

A Drosophila CRISPR/Cas9 toolkit for conditionally manipulating gene expression in the prothoracic gland as a test case for polytene tissues

Targeting gene function with spatial or temporal specificity is a key goal in molecular genetics. CRISPR-Cas9 has greatly facilitated this strategy, but some standard approaches are problematic. For instance, simple tissue-specific or global overexpression of Cas9 can cause significant lethality or developmental delays even in the absence of gRNAs. In particular, we found that Gal4-mediated expression of UAS-Cas9 in the Drosophila prothoracic gland (PG) was not a suitable strategy to disrupt gene expression, since Cas9 alone caused widespread lethality. The PG gland is widely used for studying endocrine gland function during animal development, but tools validating PG-specific RNAi phenotypes are lacking. Here, we present a collection of modular gateway-compatible CRISPR-Cas9 tools that allow precise modulation of target gene activity with temporal and spatial specificity. We also demonstrate that Cas9 fused to the progesterone ligand-binding domain can be used to activate gene expression via RU486. Using these approaches, we were able to avoid the lethality associated with simple GAL4-mediated overexpression of Cas9 in the PG. Given that the PG is a polytene tissue, we conclude that these tools work effectively in endoreplicating cells where Cas9 has to target multiple copies of the same locus. Our toolkit can be easily adapted for other tissues and can be used both for gain-and loss-of-function studies.

genetics↗

bioRxiv2018

Variation and diversification of the microbiome of Schlechtendalia chinensis on two alternate host plants

Schlechtendalia chinensis, a gall-inducing aphid, has two host plants in its life cycle. Its wintering host is a moss (typically Plagiomnium maximoviczii) and its main host is Rhus chinensis (Sumac), on which it forms galls during the summer.. This study investigated bacteria associated with S. chinensis living on the two different host plants by sequencing 16S rRNAs. A total of 183 Operational Taxonomic Units (OTUs) from 50 genera were identified from aphids living on moss, whereas 182 OTUs from 49 genera were found from aphids living in Sumac galls. The most abundant bacterial genus among identified OTUs from aphids feeding on both hosts was Buchnera. Despite similar numbers of OTUs, the composition of bacterial taxa showed significant differences between aphids living on moss and those living on R. chinensis. Specifically, there were 12 OTUs from 5 genera (family) unique to aphids living on moss, and 11 OTUs from 4 genera (family) unique to aphids feeding in galls on R. Chinensis. Principal Coordinate Analysis (PCoA) also revealed that bacteria from moss-residing aphids clustered differently from aphids collected from galls. Our results provid a foundation for future analyses on the roles of symbiotic bacteria in plant - aphid interactions in general, and how gall-specific symbionts differ in this respect.

microbiology↗