Effect of Ephestia kuehniella Eggs on Development and Transcriptome of the Ladybird Beetle Propylea japonica
by
, and
Guannan Li
1, 
Pei-Tao Chen
1,
Mei-Lan Chen
1,
Tuo-Yan Chen
1,
Yu-Hao Huang
1,
Xin Lü
2,
Hao-Sen Li
1,*
and
Hong Pang
1,*
1
State Key Laboratory of Biocontrol, School of Ecology, Sun Yat-sen University, Shenzhen 518000, China
2
Guangdong Key Laboratory of Animal Conservation and Resource Utilization, Guangdong Public Laboratory of Wild Animal Conservation and Utilization, Institute of Zoology, Guangdong Academy of Sciences, Guangzhou 510275, China
*
Authors to whom correspondence should be addressed.
Insects 2024, 15(6), 407; https://doi.org/10.3390/insects15060407
Submission received: 18 April 2024
/
Revised: 26 May 2024
/
Accepted: 31 May 2024
/
Published: 2 June 2024
(This article belongs to the Special Issue Genetics and Evolution of Ladybird Beetles in Biological Control)
Abstract
:Simple Summary
Improving augmentative biological control relies on the development of a cost-effective and readily available diet for rearing natural enemy insects. This study highlights the effectiveness of Ephestia kuehniella eggs in promoting the development of the predatory ladybird beetle Propylea japonica. It represents the first study in assessing the advantages of E. kuehniella eggs as dietary sources for ladybirds by examining transcriptional regulation.
Abstract
The eggs of the Mediterranean flour moth, Ephestia kuehniella, are frequently utilized as alternative diets and have demonstrated promising outcomes when consumed by various insects. Nonetheless, the specific reasons for their effectiveness remain unclear. In our study, we assessed the developmental performance of the ladybird Propylea japonica when fed E. kuehniella eggs, alongside 12 factitious prey or artificial diets. Our findings revealed that ladybirds fed E. kuehniella eggs displayed a performance comparable to those fed the natural prey Megoura crassicauda. Transcriptome profiling of larvae raised on E. kuehniella eggs and M. crassicauda revealed that genes upregulated in the former group were enriched in metabolic pathways associated with carbohydrates, lipids, and other essential nutrients. This suggests that E. kuehniella eggs may have a higher nutrient content compared to natural prey. Furthermore, a notable downregulation in the expression of immune effector genes, such as Attacin and Coleoptericin, was observed, which might be attributed to the lower microbial content in E. kuehniella eggs compared to M. crassicauda. We suggest that the difference between E. kuehniella eggs and M. crassicauda as food sources for P. japonica lies in their nutrient and microbial contents. These findings provide valuable insights for the advancement of innovative artificial breeding systems for natural enemies.
1. Introduction
Augmentative biological control stands as an important method in agricultural pest management, involving the release of large quantities of natural enemies into the field [1,2]. The production of these natural enemies relies on a three-tiered artificial rearing system encompassing predators, prey, and the prey’s host plants. Nonetheless, challenges such as discontinuity and high rearing facility costs can impede this method’s efficacy [3,4]. Thus, the development of a cost-effective and easily accessible artificial diet is crucial for enhancing biological control strategies.
The eggs of the Mediterranean flour moth, Ephestia kuehniella Zeller, 1879 (Lepidoptera, Pyralidae), are widely used as a factitious diet [5]. They offer the advantage of being easily obtainable and preservable. These eggs are reported to contain higher levels of nutrients, with three times more amino acids and lipids than aphids like Acyrthosiphon pisum (Harris, 1776) (Hemiptera, Aphididae) [5]. These eggs have been successfully utilized to feed various predatory ladybirds such as Adalia bipunctata (Linnaeus, 1758) (Coleoptera, Coccinellidae), Harmonia axyridis (Pallas, 1773) (Coleoptera, Coccinellidae), and Cryptolaemus montrouzieri Mulsant, 1853 (Coleoptera, Coccinellidae) [4,6,7,8,9,10]. They have been shown to enhance growth and developmental rates compared to natural prey options [5,7,10] and also serve as hosts for parasitoid natural enemies like Trichogramma (Hymenoptera, Trichogrammatidae) [11]. However, the suitability of E. kuehniella eggs varies among different ladybirds, as they are not suitable for rearing Coccinella septempunctata (Linnaeus, 1758) (Coleoptera, Coccinellidae) [6,12]. The specific reasons for the effectiveness of E. kuehniella eggs as ladybird diets are still largely unknown.
Predatory ladybirds are commonly used as natural enemies in augmentative biological control due to their pest-feeding abilities [4]. Propylea japonica (Thunberg, 1781) (Coleoptera, Coccinellidae), a native of Asia, predominantly preys on aphids and various agricultural pests. Known for its adaptability to diverse diet regimes and temperature ranges, P. japonica are widely used in augmentative biological control [13]. This study aimed to assess the suitability of E. kuehniella eggs as a diet for P. japonica and explore the differences between E. kuehniella eggs and natural prey as food sources for this ladybird. We compared the life history traits of P. japonica when fed on 14 diets, which included natural prey like Megoura crassicauda Mordvilko, 1919 (Hemiptera, Aphididae) aphids, factitious prey such as E. kuehniella eggs, and artificial diets. We also performed transcriptome profiling to identify differentially expressed genes (DEGs) in P. japonica in response to feeding on E. kuehniella eggs.
2. Materials and Methods
2.1. Preparing of the Ladybird and Diets
The ladybird P. japonica in the experiment was collected from Baiyun Mountain, Guangzhou, China (113°17′ E, 23°11′ N) between 2014 and 2018. Before the experiment, P. japonica eggs were harvested from the cages using plastic straws (0.5 cm in diameter, 3 cm in length) and transferred to plastic Petri dishes (15 cm in diameter, 1.5 cm in height). The hatched larvae from these eggs were then used in the study. A total of 14 diet treatments (Table 1) were assessed for the rearing P. japonica. Nine diet materials were formulated into a culture medium based on a previously published recipe [14], comprising 2.5 g of sucrose, 1.5 g of a protein source, 0.8 g of agarose, and 10 mL of distilled water. The eggs of E. kuehniella (FLO) and Spodoptera frugiperda (FAL) were stored in a freezer at −20 °C and thawed at 4 °C before use.
2.2. Feeding Experiments of P. japonica on Different Diets
The feeding experiments were performed from March, 2017 to December, 2023. Newly emerged male and female adults were paired in plastic Petri dishes (9 cm in diameter, 1.5 cm in height), and the eggs they laid were collected. The newly hatched larvae were individually placed into plastic Petri dishes (3 cm in diameter, 1 cm in height) and divided into 14 groups with varying diets. Each dish was equipped with a moist cotton to ensure water availability. Throughout the experiment, the total developmental time, adult weight, and survival rate were documented daily, with daily replenishment of food. Adults were weighed on the day of emergence. The experiment included a total of 14 treatments and 24 batches (Table 1). To evaluate the possible batch effect of the main treatments APH and FLO, three batches were conducted for each. For the other treatments, one to three batches were conducted. Each batch comprised 22–99 tested individuals (Table 1). All feeding experiments were performed in climatic chambers at 25 (±1) °C, 75 (±5) % RH under a 16:8 (L: D) photoperiod. Statistical analyses for comparing life history traits among different treatments and batches were carried out using R software v4.2.3. The Shapiro–Wilk normality test and Bartlett’s test were employed to assess the normality and homogeneity of variance for the developmental time and adult weight data. Data conforming to these assumptions underwent one-way ANOVA analysis. In cases where a significant difference was observed, Tukey’s Honestly Significant Difference (HSD) test was utilized for post hoc comparisons. Alternatively, the Wilcoxon signed-rank test was applied. A significance level of p < 0.05 was adopted for all statistical tests.
2.3. Transcriptome Analysis
Fourth-instar larvae (<24 h after molting) of P. japonica from the above batches APH01 and FLO02 were randomly collected for transcriptome analysis. An individual larva was used as a single biological replicate, and three and two biological replicates were set for APH01 and FLO02, respectively. The total RNA of each individual was extracted using TRIzol reagent (CWBIO, Beijing, China). RNA quality and quantity were determined using a Nanodrop 1000 spectrophotometer (Thermo Fisher Scientific, Wilmington, NC, USA). Only RNA samples with a 260/280 ratio from 1.8 to 2.0, a 260/230 ratio from 2.0 to 2.5, and an RNA integrity number (RIN) > 8.0 were used for sequencing. Sequencing was performed on the Illumina HiSeq 2500 platform. Adaptors and low-quality sequences were removed using the default settings for Trimmomatic v0.36 [15]. The clean reads were then mapped to the published genome of P. japonica [16] using HISAT2 v2.2.1 [17]. Abundance estimation was performed using StringTie v2.1.4 [18] based on genome annotation. Differential expression analysis between treatments was performed using DESeq2 [19] according to the standard workflow, with a log2(fold change) (log2FC) value > 1 or <−1 and an adjusted p value (Q value) < 0.05 used as the criteria for defining differentially expressed genes (DEGs). Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway annotations were performed using eggnog-mapper v2 (eggNOG database v5.0) [20,21]. Functional enrichment of DEGs was performed in the R package clusterProfiler [22]. Visualization of GO enrichment was performed using GO-Figure! [23].
3. Results
3.1. The Analysis of Life History Traits of P. japonica on Different Diet
In the analysis of life history traits, 9 out of the 14 treatments (APH, FLO, PUP, HON, WHI, POL, BRI, FAL, and YEL) led to individuals reaching the adult stage (Figure 1A). The survival rates of APH were generally lower than those of FLO (Figure 1A). Treatments WHI, POL, BRI, FAL, and YEL had less than 25% of the tested individuals reaching adulthood (Figure 1A), and their developmental time and adult weight data were excluded from further analysis. Figure 1B integrated the parameters of survival rate, developmental time, and adult weight, demonstrating that APH and FLO exhibited superior performance compared to PUP and HON. None of the life history traits were significantly different between the APH and FLO treatments (Table 2). In addition, the performance of APH and FLO exhibited variability among batches. For example, the development time of a batch in FLO (FLO02) was significantly shorter than not only batches in APH (APH02–03) but also FLO (FLO01) (p < 0.05 in Wilcoxon signed-rank test) (Figure 1C). Similarly, the adult weight of a batch in APH (APH03) was significantly higher than batches in both APH (APH02) and FLO (FLO02–03) (p < 0.05 in Wilcoxon signed-rank test) (Figure 1D).
3.2. The Analysis of Transcriptome Profiling between APH and FLO
In the comparative analysis of transcriptome profiling between APH (as control) and FLO (as treatment), the samples from the treatment group displayed coefficient of determination (r2) values ranging from 0.82 to 0.93, with a mean of 0.88 (Table 3). A total of 479 differentially expressed genes were identified, with 345 genes upregulated and 134 genes downregulated. Specifically, immune effector genes such as Attacin and Coleoptericin were significantly downregulated (Figure 2). The results of the Gene Ontology (GO) enrichment analysis revealed that the significantly upregulated genes were mainly enriched in nutritional metabolic functions, including catalytic activity, transmembrane transporter activity in the Molecular Function (MF) category, and carbohydrate metabolic processes in the Biological Process (BP) category (Figure 3). Conversely, the downregulated genes were only enriched in two GO terms: 4-hydroxyphenylpyruvate dioxygenase activity and tyrosine catabolic process. The Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis showed similar findings, with upregulated genes being enriched in pathways related to glycogen metabolism (Table 4).
4. Discussion
In the feeding experiment, most of the artificial diets resulted in a poorer performance of P. japonica than the natural prey pea aphids M. crassicauda. Some diets could only support a few ladybirds to mature into adults, such as whiteflies (WHI treatment) and the pollen of Brassica campestris (POL treatment). Other diets, like the pupae of the honeybee Apis mellifera (PUP treatment) and bee honey (HON treatment), could sustain over 50% of larvae to reach the adult stage, but at the cost of a significantly decreased larval development time and adult weight. Hence, none of these diets were deemed suitable for P. japonica. Among all the tested artificial diets, the natural prey pea aphids M. crassicauda (APH treatment) and E. kuehniella eggs (FLO treatment) exhibited the best performance for feeding P. japonica. Although there was variability in performance among batches in our experiments, the life history traits of P. japonica reared on both E. kuehniella eggs and aphids showed relatively similar patterns. This finding aligns with previous reports that identified E. kuehniella eggs as a viable alternative diet for the development and reproduction of P. japonica [8]. Therefore, E. kuehniella eggs are suitable for use as an alternative food source in the artificial feeding system of P. japonica.
By comparing the transcriptome of P. japonica fed on E. kuehniella eggs with that of aphids, we identified numerous upregulated metabolism-related genes that were enriched in several sugar metabolism pathways. This outcome parallels findings in previous studies on other ladybird species, indicating that changes in life history traits and transcriptome regulation may reflect variations in the nutritional composition of the E. kuehniella eggs [14,24]. Similar changes in life history traits and differentially expressed metabolism-related genes in response to varied nutritional compositions of diets have been observed in other insects. For example, nutrient-sensing and metabolic pathways in A. mellifera are activated by feeding on honey, which contains higher levels of proteins and amino acids, compared to sugar feeding [25]. These results underscore the importance of nutrient and biochemical composition in assessing the suitability of an artificial diet [26]. However, we did not test the nutritional content differences between E. kuehniella eggs and pea aphids M. crassicauda. Additionally, we cannot rule out the possibility that other factors also contributed to the differential expression of metabolism-related genes.
Aphids are known to harbor various symbiotic bacteria, such as Buchnera aphidicola, Serratia symbiotica, Hamiltonella defensa, Regiella insecticola, Rickettsia, Rickettsiella, Spiroplasma, Wolbachia, and Arsenophonus [27]. Some of these bacteria may have adverse effects on the natural enemies to protect their host [28,29]. These bacteria can change the gene expression in the natural enemies [30]. In contrast, E. kuehniella eggs contain fewer microbes that could elicit changes in gene expression in ladybirds. Previous studies have reported that the >99% microbiota composition of these eggs belonged to an intracellular bacterium, Wolbachia [24]. In this study, two immune effector genes of P. japonica were significantly downregulated when feeding on E. kuehniella eggs, suggesting that these genes may play a crucial role in eliminating bacteria from aphids. Therefore, feeding on E. kuehniella eggs may result in less immune stress, contributing to the superior performance of P. japonica feeding on this diet.
Our research highlights that E. kuehniella eggs are a suitable artificial diet for the mass-rearing of P. japonica, surpassing other tested alternative prey or artificial diets. Through the transcriptome profiling analysis, we suggest that the difference between E. kuehniella eggs and M. crassicauda as food sources for P. japonica lies in their nutrient and microbial contents. These findings also offer valuable insights for developing improved systems for the captive breeding of other natural enemies. However, the effects of feeding E. kuehniella eggs to multiple successive generations of P. japonica remain unclear and need further research for clarification.
Author Contributions
Conceptualization, G.L., P.-T.C., M.-L.C., T.-Y.C., Y.-H.H., X.L., H.-S.L. and H.P.; methodology, G.L. and H.-S.L.; software, Y.-H.H. and H.-S.L.; validation, G.L., H.-S.L. and H.P.; formal analysis, G.L., H.-S.L. and H.P.; investigation, G.L., P.-T.C., M.-L.C. and T.-Y.C.; resources, X.L., H.-S.L. and H.P.; data curation, Y.-H.H. and H.-S.L.; writing—original draft preparation, G.L., P.-T.C. and H.-S.L.; writing—review and editing, G.L., P.-T.C., M.-L.C., T.-Y.C., Y.-H.H., X.L., H.-S.L. and H.P.; visualization, G.L., P.-T.C. and H.-S.L.; supervision, H.-S.L. and H.P.; project administration, X.L., H.-S.L. and H.P.; funding acquisition, X.L., H.-S.L. and H.P. All authors have read and agreed to the published version of the manuscript.
Funding
This work was supported by the National Natural Science Foundation of China (Grant No. 32172472), the Open Fund of Guangdong Key Laboratory of Animal Protection and Resource Utilization (Grant No. GIZ-KE202304), and the National Key R&D Program of China (Grant No. 2023YFD1400600).
Data Availability Statement
The raw reads of the transcriptome sequencing were deposited in NCBI under the project of PRJNA956151 (SRR24183495–SRR24183499).
Conflicts of Interest
The authors declare no conflicts of interest.
References
- De Clercq, P.; Mason, P.G.; Babendreier, D. Benefits and risks of exotic biological control agents. BioControl 2011, 56, 681–698. [Google Scholar] [CrossRef]
- van Lenteren, J.C. The state of commercial augmentative biological control: Plenty of natural enemies, but a frustrating lack of uptake. BioControl 2012, 57, 1–20. [Google Scholar] [CrossRef]
- Riddick, E.W. Benefits and limitations of factitious prey and artificial diets on life parameters of predatory beetles, bugs, and lacewings: A mini-review. BioControl 2008, 54, 325–339. [Google Scholar] [CrossRef]
- Sun, Y.-X.; Hao, Y.-N.; Riddick, E.W.; Liu, T.-X. Factitious prey and artificial diets for predatory lady beetles: Current situation, obstacles, and approaches for improvement: A review. Biocontrol Sci. Technol. 2017, 27, 601–619. [Google Scholar] [CrossRef]
- Specty, O.; Febvay, G.; Grenier, S.; Delobel, B.; Piotte, C.; Pageaux, J.F.; Ferran, A.; Guillaud, J. Nutritional plasticity of the predatory ladybeetle Harmonia axyridis (Coleoptera: Coccinellidae): Comparison between natural and substitution prey. Arch. Insect Biochem. 2003, 52, 81–91. [Google Scholar] [CrossRef] [PubMed]
- Mercer, N.H.; Obrycki, J. Impacts of larval diet on pre-imaginal development, survival and adult size of six species of Coccinellidae (Coleoptera). J. Kans. Entomol. Soc. 2021, 93, 256–261. [Google Scholar] [CrossRef]
- De Clercq, P.; Bonte, M.; Van Speybroeck, K.; Bolckmans, K.; Deforce, K. Development and reproduction of Adalia bipunctata (Coleoptera: Coccinellidae) on eggs of Ephestia kuehniella (Lepidoptera: Phycitidae) and pollen. Pest Manag. Sci. 2005, 61, 1129–1132. [Google Scholar] [CrossRef] [PubMed]
- Hamasaki, K.; Matsui, M. Development and reproduction of an aphidophagous coccinellid, Propylea japonica (Thunberg) (Coleoptera: Coccinellidae), reared on an alternative diet, Ephestia kuehniella Zeller (Lepidoptera: Pyralidae) eggs. Appl. Entomol. Zool. 2006, 41, 233–237. [Google Scholar] [CrossRef]
- dos Santos Rodrigues, A.R.; Spíndola, A.F.; de Morais Oliveira, J.E.; Torres, J.B. Dietary effects upon biological performance and lambda-cyhalothrin susceptibility in the multicolored Asian lady beetle, Harmonia axyridis. J. Phytoparasitica 2013, 41, 285–294. [Google Scholar] [CrossRef]
- Maes, S.; Grégoire, J.-C.; De Clercq, P. Prey range of the predatory ladybird Cryptolaemus montrouzieri. BioControl 2014, 59, 729–738. [Google Scholar] [CrossRef]
- St-Onge, M.; Cormier, D.; Todorova, S.; Lucas, E. Conservation of Ephestia kuehniella eggs as hosts for Trichogramma ostriniae. J. Appl. Entomol. 2016, 140, 218–222. [Google Scholar] [CrossRef]
- Du, X.Y.; Yang, H.Y.; Gong, S.R.; Zhang, P.F.; Chen, P.T.; Liang, Y.S.; Huang, Y.H.; Tang, X.F.; Chen, Q.K.; De Clercq, P.; et al. Aphidophagous ladybird beetles adapt to an aphid symbiont. Funct. Ecol. 2022, 36, 2593–2604. [Google Scholar] [CrossRef]
- Pervez, A. Ecology of aphidophagous ladybird Propylea species: A review. J. Asia-Pac. Entomol. 2011, 14, 357–365. [Google Scholar] [CrossRef]
- Li, H.-S.; Huang, Y.-H.; Chen, M.-L.; Ren, Z.; Qiu, B.-Y.; De Clercq, P.; Heckel, G.; Pang, H. Genomic insight into diet adaptation in the biological control agent Cryptolaemus montrouzieri. BMC Genom. 2021, 22, 135. [Google Scholar] [CrossRef] [PubMed]
- Bolger, A.M.; Lohse, M.; Usadel, B. Trimmomatic: A flexible trimmer for Illumina sequence data. Bioinformatics 2014, 30, 2114–2120. [Google Scholar] [CrossRef]
- Zhang, L.; Li, S.; Luo, J.; Du, P.; Wu, L.; Li, Y.; Zhu, X.; Wang, L.; Zhang, S.; Cui, J. Chromosome-level genome assembly of the predator Propylea japonica to understand its tolerance to insecticides and high temperatures. Mol. Ecol. Resour. 2019, 20, 292–307. [Google Scholar] [CrossRef] [PubMed]
- Kim, D.; Landmead, B.; Salzberg, S.L. HISAT: A fast spliced aligner with low memory requirements. Nat. Methods 2015, 12, 357–360. [Google Scholar] [CrossRef]
- Kovaka, S.; Zimin, A.V.; Pertea, G.M.; Razaghi, R.; Salzberg, S.L.; Pertea, M. Transcriptome assembly from long-read RNA-seq alignments with StringTie2. Genome Biol. 2019, 20, 278. [Google Scholar] [CrossRef]
- Love, M.I.; Huber, W.; Anders, S. Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2. Genome Biol. 2014, 15, 550. [Google Scholar] [CrossRef]
- Huerta-Cepas, J.; Szklarczyk, D.; Heller, D.; Hernandez-Plaza, A.; Forslund, S.K.; Cook, H.; Mende, D.R.; Letunic, I.; Rattei, T.; Jensen, L.J.; et al. eggNOG 5.0: A hierarchical, functionally and phylogenetically annotated orthology resource based on 5090 organisms and 2502 viruses. Nucleic Acids Res. 2019, 47, D309–D314. [Google Scholar] [CrossRef]
- Cantalapiedra, C.P.; Hernandez-Plaza, A.; Letunic, I.; Bork, P.; Huerta-Cepas, J. eggNOG-mapper v2: Functional annotation, orthology assignments, and domain prediction at the metagenomic scale. Mol. Biol. Evol. 2021, 38, 5825–5829. [Google Scholar] [CrossRef]
- Yu, G.; Wang, L.G.; Han, Y.; He, Q.Y. clusterProfiler: An R package for comparing biological themes among gene clusters. OMICS 2012, 16, 284–287. [Google Scholar] [CrossRef] [PubMed]
- Reijnders, M.J.M.F.; Waterhouse, R.M. Summary Visualizations of Gene Ontology Terms with GO-Figure! Front. Bioinform. 2021, 1, 638255. [Google Scholar] [CrossRef] [PubMed]
- Huang, Y.-H.; Du, X.-Y.; Chen, P.-T.; Tang, X.-F.; Gong, S.-R.; Zhang, P.-F.; Yang, H.-Y.; De Clercq, P.; Li, H.-S.; Pang, H. Is pollinivory in the omnivorous ladybird beetle Micraspis discolor (Coleoptera: Coccinellidae) symbiosis-dependent? Biol. Control 2022, 169, 104867. [Google Scholar] [CrossRef]
- Alaux, C.; Dantec, C.; Parrinello, H.; Le Conte, Y. Nutrigenomics in honey bees: Digital gene expression analysis of pollen’s nutritive effects on healthy and varroa-parasitized bees. BMC Genom. 2011, 12, 496. [Google Scholar] [CrossRef] [PubMed]
- Piper, M.D. Using artificial diets to understand the nutritional physiology of Drosophila melanogaster. Curr. Opin. Insect Sci. 2017, 23, 104–111. [Google Scholar] [CrossRef] [PubMed]
- Guo, J.; Hatt, S.; He, K.; Chen, J.; Francis, F.; Wang, Z. Nine facultative endosymbionts in aphids. A review. J. Asia-Pac. Entomol. 2017, 20, 794–801. [Google Scholar] [CrossRef]
- Haine, E.R. Symbiont-mediated protection. Proc. R. Soc. B Biol. Sci. 2007, 275, 353–361. [Google Scholar] [CrossRef] [PubMed]
- Brownlie, J.C.; Johnson, K.N. Symbiont-mediated protection in insect hosts. Trends Microbiol. 2009, 17, 348–354. [Google Scholar] [CrossRef]
- Dennis, A.B.; Patel, V.; Oliver, K.M.; Vorburger, C. Parasitoid gene expression changes after adaptation to symbiont-protected hosts. Evolution 2017, 71, 2599–2617. [Google Scholar] [CrossRef]
Figure 1.
Effects of different diet treatments and batches on life history traits of Propylea japonica. (A) Survival rate of batches in APH, FLO, PUP, HON, WHI, POL, BRI, FAL, and YEL treatments. (B) Survival rate (point size), development time (X-axis), and adult weight (Y-axis) of batches in APH, FLO, PUP, and HON treatments. (C) Development time and (D) female adult weight of batches in APH and FLO treatments. Different colors indicate different diet treatments. Bars with the same letter are not significantly different (p ≥ 0.05).
Figure 1.
Effects of different diet treatments and batches on life history traits of Propylea japonica. (A) Survival rate of batches in APH, FLO, PUP, HON, WHI, POL, BRI, FAL, and YEL treatments. (B) Survival rate (point size), development time (X-axis), and adult weight (Y-axis) of batches in APH, FLO, PUP, and HON treatments. (C) Development time and (D) female adult weight of batches in APH and FLO treatments. Different colors indicate different diet treatments. Bars with the same letter are not significantly different (p ≥ 0.05).
Figure 2.
Volcano plot of the transcriptome comparison in Propylea japonica when feeding on Ephestia kuehniella eggs (FLO treatment) compared to pea aphids Megoura crassicauda (APH treatment). X-axis: log2 of the fold change values. Y-axis: −log10 of the adjusted p values (Q). Red: upregulated differentially expressed genes (DEGs). Blue: downregulated DEGs. SOA: Sterol O-acyltransferase 1, RPS10: 40S ribosomal protein S10, RPL12: 60S ribosomal protein L12, LST: Luciferin sulfotransferase, AA: Alpha-amylase, TRET1: Trehalose transporter 1, ST1: Sulfotransferase 1C2, FT: Formyl transferase, POE: Protein obstructor-E, UNC93: Ion channel regulatory protein, UGT: UDP-glycosyltransferase, EBD: Estradiol 17-beta-dehydrogenase 11. Larger absolute values on the X-axis and higher values on the Y-axis indicate greater differential expression of genes.
Figure 2.
Volcano plot of the transcriptome comparison in Propylea japonica when feeding on Ephestia kuehniella eggs (FLO treatment) compared to pea aphids Megoura crassicauda (APH treatment). X-axis: log2 of the fold change values. Y-axis: −log10 of the adjusted p values (Q). Red: upregulated differentially expressed genes (DEGs). Blue: downregulated DEGs. SOA: Sterol O-acyltransferase 1, RPS10: 40S ribosomal protein S10, RPL12: 60S ribosomal protein L12, LST: Luciferin sulfotransferase, AA: Alpha-amylase, TRET1: Trehalose transporter 1, ST1: Sulfotransferase 1C2, FT: Formyl transferase, POE: Protein obstructor-E, UNC93: Ion channel regulatory protein, UGT: UDP-glycosyltransferase, EBD: Estradiol 17-beta-dehydrogenase 11. Larger absolute values on the X-axis and higher values on the Y-axis indicate greater differential expression of genes.
Figure 3.
Gene Ontology (GO) enrichment analysis of differentially expressed genes in Propylea japonica when feeding on Ephestia kuehniella eggs (FLO treatment) compared to pea aphids Megoura crassicauda (APH treatment). Each bubble represents a cluster of similar GO terms summarized by a representative term reported in the legend and sorted by the average p values of the representative GO term across the module of (A) Molecular Function and (B) Biological Process. Bubble size indicates the number of GO terms in each cluster, and the color is the average p value of the representative GO term across the gene modules. Similar clusters plot closer to each other.
Figure 3.
Gene Ontology (GO) enrichment analysis of differentially expressed genes in Propylea japonica when feeding on Ephestia kuehniella eggs (FLO treatment) compared to pea aphids Megoura crassicauda (APH treatment). Each bubble represents a cluster of similar GO terms summarized by a representative term reported in the legend and sorted by the average p values of the representative GO term across the module of (A) Molecular Function and (B) Biological Process. Bubble size indicates the number of GO terms in each cluster, and the color is the average p value of the representative GO term across the gene modules. Similar clusters plot closer to each other.
Table 1.
Information of the treatments, batches, number of individuals tested, and dates of the experiments conducted in this study.
Table 1.
Information of the treatments, batches, number of individuals tested, and dates of the experiments conducted in this study.
| Treatment | Batch | DIET MATERIAL | Process for Diet | Individual | Date |
|---|---|---|---|---|---|
| APH | APH01 | Pea aphids Megoura crassicauda | Live prey | 65 | October 2021 |
| APH | APH02 | Pea aphids Megoura crassicauda | Live prey | 42 | Novermber 2018 |
| APH | APH03 | Pea aphids Megoura crassicauda | Live prey | 40 | September 2022 |
| FLO | FLO01 | Eggs of flour moth Ephestia kuehniella | Cold-stored | 40 | August 2021 |
| FLO | FLO02 | Eggs of flour moth Ephestia kuehniella | Cold-stored | 97 | October 2021 |
| FLO | FLO03 | Eggs of flour moth Ephestia kuehniella | Cold-stored | 56 | May 2021 |
| PUP | PUP01 | Pupae of honeybee Apis mellifera | Dry powder and solid medium | 22 | March 2017 |
| PUP | PUP02 | Pupae of honeybee Apis mellifera | Dry powder and solid medium | 50 | March 2017 |
| HON | HON01 | Bee honey | Solid medium | 62 | Novermber 2018 |
| HON | HON02 | Bee honey | Solid medium | 22 | March 2017 |
| WHI | WHI01 | Whitefies Bemisia tabaci | Live prey | 50 | Novermber 2018 |
| POL | POL01 | Pollen of Brassica campestris | Plant materials | 99 | Novermber 2023 |
| BRI | BRI01 | Cysts of brine shrimp Artemia salina | Solid medium | 69 | Novermber 2018 |
| BRI | BRI02 | Cysts of brine shrimp Artemia salina | Solid medium | 47 | March 2017 |
| FAL | FAL01 | Eggs of fall armyworm Spodoptera frugiperda | Cold-stored | 28 | December 2023 |
| YEL | YEL01 | Larvae of yellow mealworm Tenebrio molitor | Dry powder and solid medium | 88 | Novermber 2018 |
| YEL | YEL02 | Larvae of yellow mealworm Tenebrio molitor | Dry powder and solid medium | 23 | March 2017 |
| YEL | YEL03 | Larvae of yellow mealworm Tenebrio molitor | Dry powder and solid medium | 29 | March 2017 |
| BLA | BLA01 | Larvae of black soldier fly Hermetia illucens | Dry powder and solid medium | 28 | March 2017 |
| BLA | BLA02 | Larvae of black soldier fly Hermetia illucens | Dry powder and solid medium | 30 | May 2021 |
| SIL | SIL01 | Pupae of silkworm Bombyx mori | Dry powder and solid medium | 74 | March 2017 |
| POR | POR01 | Pork | Dry powder and solid medium | 20 | March 2017 |
| LIV | LIV01 | Pork liver | Dry powder and solid medium | 18 | March 2017 |
| CHI | CHI01 | Chicken egg | Solid medium | 25 | March 2017 |
Table 2.
Comparison of life history traits in Propylea japonica when feeding on Ephestia kuehniella eggs (FLO treatment) compared to pea aphids Megoura crassicauda (APH treatment).
Table 2.
Comparison of life history traits in Propylea japonica when feeding on Ephestia kuehniella eggs (FLO treatment) compared to pea aphids Megoura crassicauda (APH treatment).
| APH | FLO | Wilcoxon Signed-Rank Test | |
|---|---|---|---|
| Survival rate (%) | 63.792 ± 3.541 | 84.277 ± 2.926 | Not significant |
| Development time (day) | 12.017 ± 0.226 | 11.583 ± 0.305 | Not significant |
| Female adult weight (mg) | 6.693 ± 0.621 | 6.243 ± 0.289 | Not significant |
| Male adult weight (mg) | 5.484 ± 0.308 | 5.247 ± 0.307 | Not significant |
Table 3.
Coefficient of determination (r2) values of gene expression between the studied transcriptomes.
Table 3.
Coefficient of determination (r2) values of gene expression between the studied transcriptomes.
| APH | APH | APH | FLO | FLO | |
|---|---|---|---|---|---|
| APH | |||||
| APH | 0.890 | ||||
| APH | 0.910 | 0.922 | |||
| FLO | 0.862 | 0.942 | 0.908 | ||
| FLO | 0.820 | 0.821 | 0.874 | 0.889 |
Table 4.
Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis of differentially expressed genes in Propylea japonica when feeding on Ephestia kuehniella eggs (FLO treatment) compared to pea aphids Megoura crassicauda (APH treatment).
Table 4.
Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis of differentially expressed genes in Propylea japonica when feeding on Ephestia kuehniella eggs (FLO treatment) compared to pea aphids Megoura crassicauda (APH treatment).
| Regulation | ID | Descrption | Q Value | Ratio |
|---|---|---|---|---|
| up | ko00500 | Metabolism: Starch and sucrose metabolism | 7.30 × 10−15 | 16/59 |
| up | ko01100 | Metabolism: Metabolic pathways | 2.44 × 10−9 | 69/2498 |
| up | ko00052 | Metabolism: Galactose metabolism | 9.28 × 10−7 | 11/83 |
| up | ko01200 | Metabolism: Carbon metabolism | 1.47 × 10−4 | 14/231 |
| up | ko00010 | Metabolism: Glycolysis/Gluconeogenesis | 5.78 × 10−4 | 9/107 |
| up | ko00830 | Metabolism: Retinol metabolism | 0.0069 | 9/153 |
| up | ko00310 | Metabolism: Lysine degradation | 0.0069 | 8/121 |
| up | ko00980 | Metabolism: Metabolism of xenobiotics by cytochrome P450 | 0.0095 | 9/163 |
| up | ko00053 | Metabolism: Steroid hormone biosynthesis | 0.0134 | 8/143 |
| up | ko00650 | Metabolism: Ascorbate and aldarate metabolism | 0.0148 | 4/32 |
| up | ko00982 | Metabolism: Butanoate metabolism | 0.0162 | 8/151 |
| up | ko00040 | Metabolism: Drug metabolism—cytochrome P450 | 0.0167 | 8/155 |
| up | ko00071 | Metabolism: Pentose and glucuronate interconversions | 0.0167 | 6/88 |
| up | ko00520 | Metabolism: Fatty acid degradation | 0.0350 | 6/103 |
| up | ko00030 | Metabolism: Amino sugar and nucleotide sugar metabolism | 0.0377 | 4/45 |
| up | ko00983 | Metabolism: Pentose phosphate pathway | 0.0440 | 9/228 |
| up | ko00860 | Metabolism: Drug metabolism—other enzymes | 0.0447 | 7/149 |
| down | ko00061 | Metabolism: Porphyrin and chlorophyll metabolism | 0.0020 | 5/87 |
| down | ko04910 | Metabolism: Fatty acid biosynthesis | 0.0115 | 6/247 |
| down | ko01100 | Organismal Systems: Insulin signaling pathway | 0.0115 | 20/2498 |
| down | ko01212 | Metabolism: Metabolic pathways | 0.0115 | 5/176 |
| down | ko04614 | Metabolism: Fatty acid metabolism | 0.0115 | 3/43 |
| down | ko04152 | Organismal Systems: Renin-angiotensin system | 0.0137 | 5/191 |
| down | ko04640 | Environmental Information Processing: AMPK signaling pathway | 0.0284 | 2/18 |
Disclaimer/Publisher’s Note: The statements, opinions and data contained in all publications are solely those of the individual author(s) and contributor(s) and not of MDPI and/or the editor(s). MDPI and/or the editor(s) disclaim responsibility for any injury to people or property resulting from any ideas, methods, instructions or products referred to in the content. |
© 2024 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
Share and Cite
MDPI and ACS Style
Li, G.; Chen, P.-T.; Chen, M.-L.; Chen, T.-Y.; Huang, Y.-H.; Lü, X.; Li, H.-S.; Pang, H. Effect of Ephestia kuehniella Eggs on Development and Transcriptome of the Ladybird Beetle Propylea japonica. Insects 2024, 15, 407. https://doi.org/10.3390/insects15060407
AMA Style
Li G, Chen P-T, Chen M-L, Chen T-Y, Huang Y-H, Lü X, Li H-S, Pang H. Effect of Ephestia kuehniella Eggs on Development and Transcriptome of the Ladybird Beetle Propylea japonica. Insects. 2024; 15(6):407. https://doi.org/10.3390/insects15060407
Chicago/Turabian StyleLi, Guannan, Pei-Tao Chen, Mei-Lan Chen, Tuo-Yan Chen, Yu-Hao Huang, Xin Lü, Hao-Sen Li, and Hong Pang. 2024. "Effect of Ephestia kuehniella Eggs on Development and Transcriptome of the Ladybird Beetle Propylea japonica" Insects 15, no. 6: 407. https://doi.org/10.3390/insects15060407
APA StyleLi, G., Chen, P. -T., Chen, M. -L., Chen, T. -Y., Huang, Y. -H., Lü, X., Li, H. -S., & Pang, H. (2024). Effect of Ephestia kuehniella Eggs on Development and Transcriptome of the Ladybird Beetle Propylea japonica. Insects, 15(6), 407. https://doi.org/10.3390/insects15060407
Note that from the first issue of 2016, this journal uses article numbers instead of page numbers. See further details here.
