Comparative Biochemistry and Physiology Part D: Genomics and Proteomics
Comparative transcriptome analysis reveals a potential mechanism for host nutritional manipulation after parasitization by Leptopilina boulardi
Graphical abstract
Potential mechanical model of host nutritional regulation manipulated by L. boulardi.
Introduction
Insects are the most abundant and diverse group of animals on Earth. Although their feeding strategies are varied and include herbivory, carnivory, and hematophagy, the intestinal tract is the principle site for the digestion of food and the subsequent absorption of vital nutrients (Stainier, 2005; Karasov and Douglas, 2013). The midgut physiology and functions in various insect species have been well documented. For instance, the digestion of food requires the sequential and coordinated action of enzymes and nutrient transporters in the midgut (Lemaitre and Miguel-Aliaga, 2013; Buchon et al., 2013b). Most digestive enzymes are responsible for protein, carbohydrate and lipid digestion, which lead to the release of small organic nutrients from complex carbohydrates and macromolecules consumed during feeding, such as monosaccharides, monoglycerides, fatty acids, and amino acids (Buchon et al., 2013b; Miguel-Aliaga et al., 2018). These small nutrients can then be successfully absorbed by gut epithelium cells and allocated to storage or direct energy supply. Moreover, the insect gut also serves as an important barrier for the maintenance of a healthy microbe community, which is also necessary to ensure an appropriate nutrient metabolism for the development and survival of the host insects (Davis and Engström, 2012; Engel and Moran, 2013; Buchon et al., 2013a).
Parasitoid wasps are valuable biological control agents that are used to suppress the populations of pest hosts. Endoparasitoids lay eggs inside the body of their hosts, and their hatched offspring larvae acquire nutrients directly from the hosts during all of their immature stages and finally kill the hosts (Beckage and Gelman, 2004; Rajendran et al., 2017). For successful parasitization, parasitoids need to overcome the host resistance responses to protect their developing offspring. Many studies have shown that parasitoids inject several maternal factors together with their eggs during the oviposition stage, and these factors include venom, polydnaviruses (PDVs) or virus-like particles (VLPs), which help disrupt the host defense mechanism (Asgari and Rivers, 2011; Strand, 2014; Manzoor et al., 2016; Wang et al., 2018). Moreover, parasitization has often been found to manipulate the nutritional metabolism of the host to meet specific nutritional requirements (Pennacchio et al., 2014; Toledo et al., 2016; Wang et al., 2020, p.). Large-scale transcriptome analyses between parasitized and nonparasitized hosts have revealed some important genes involving in metabolic pathways, immune responses, and development, whose expression are either upregulated or downregulated during parasitization (Schlenke et al., 2007; Martinson et al., 2014). The identification of genes can help researchers explore the mechanisms through which parasitization impairs host immune responses and nutritional metabolism. The studies conducted to date have mainly focused on two tissues in parasitized hosts: circulating hemocytes and fat bodies (Boutros et al., 2002; Schlenke et al., 2007; Barandoc and Kim, 2009). Hemocytes are responsible for cellular immune defense due to parasitism, whereas the fat body is the main tissue where the host humoral response and energy mobilization occurs (Krzemień et al., 2007; Vanha-aho et al., 2015; Anderl et al., 2016). However, whether parasitoids (nonintestinal parasites) manipulate the physiological changes in the host midgut is largely unknown, even though the midgut is the most important place for food digestion and nutrient absorption and determines the nutrient quantity and quality in the whole body of the host for developing wasp offspring.
Leptopilina boulardi is a highly specialized larval-pupal parasitoid that parasitizes D. melanogaster 2nd instar larvae (Carton et al., 1986). The parasitic success of L. boulardi is ensured by venom, which is the only reported and well-studied virulence factor to date and contains 200-nm venosomes and possibly some other virulence factors (Dubuffet et al., 2009; Wan et al., 2017). The venom protein LbGAP has been found to rearrange the cytoskeleton of host hemocytes to prevent the encapsulation defense response (Labrosse et al., 2005). Recently, another venom protein, Warm, was found to be involved in the L. boulardi wasp egg attachment strategy, which is used to passively escape the host encapsulation response (Huang et al., 2021). However, less is known about how parasitization impairs host physiology in tissues other than the fat body and hemocytes. Here, we applied RNAseq to analyze the transcriptional responses of the midguts of L. boulardi-parasitized and nonparasitized D. melanogaster host larvae and attempted to identify the differentially expressed genes (DEGs) due to wasp infection. Our data will thus contribute to the elucidation of the underlying important mechanisms through which parasitization initiates changes in the host nutrient levels, which are critical for parasitoid development.
Section snippets
Transcriptomes of the host midgut after L. boulardi parasitization
To comprehensively capture the transcriptional response associated with L. boulardi parasitization in the host midgut, RNA samples were isolated from the midguts of parasitized D. melanogaster larvae at 24 and 48 h post parasitization (Fig. 1A). For comparative purposes, RNA samples were also generated from the midguts of nonparasitized D. melanogaster larvae at the same ages to serve as the controls (Fig. 1A). Four independent biological replicates of each condition were sequenced, which
Discussion
Parasitoid wasps are free-living as adults, but the offspring develop inside (endoparasitoids) or outside (ectoparasitoids) their hosts. Host nutrients act not only as primary sources of energy supply but also as regulators of growth that enable parasitoids to survive (Pennacchio et al., 2014; Rajendran et al., 2017). Recent studies have shown that parasitoids have evolved to manipulate their host nutrient levels, including sugar concentrations, to meet their dietary requirement for offspring
Insects
D. melanogaster W1118 (host) was maintained on standard cornmeal/molasses/agar medium at room temperature. Leptopilina boulardi was kindly provided by Dr. Dan Hultmark (Umeå University, Sweden) and bred on D. melanogaster W1118 since April 2016 in our lab. The wasps were raised at a temperature of 25 ± 1 °C with a relative humidity of 50–60% and a photoperiod of 16 h:8 h (L:D) inside plastic bottles (with a length of approximately 10 cm and a diameter of 5 cm). Adult wasps were provided apple
Data availability
The raw Illumina short-read sequences of all 16 samples are available through the Sequence Read Archive (SRA) under project accession PRJNA694742.
Declaration of competing interest
The authors declare that they have no competing interests.
Acknowledgments
We thank Dr. Dan Hultmark for providing parasitoid species. This research was supported by the National Key R&D Program of China (2017YFD0200400), Zhejiang Provincial Natural Science Foundation of China (No. LR18C140001), the National Science Fund for Excellent Young Scholars (31622048), the National Science Foundation of China (31772522 and 31630060).
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2022, Current Opinion in Insect ScienceCitation Excerpt :These responses include the timed regulation of various immunity genes, including pattern recognition receptors, members of the Toll, Imd and Jak/STAT immunity signal transduction pathways, several proteases and effector molecules [46,47]. Additionally, the infestation with parasitoids may alter the expression of metabolic pathways in the gut of the host larvae [48]. This may be associated with the energetic costs of launching an immune response [46,49], or it may constitute an active manipulation of parasitoids to enhance the nutritional quality of their host [48].
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