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. 2025 May 12;33(4):1567–1580. doi: 10.1111/1744-7917.70069

Food nutrition and facultative endosymbiont modulate dietary breadth of a polyphagous aphid

Chun‐Yan Chang 1, Ya‐Ni Zhao 1, Hui‐Fang Guo 1,2,✉, Xiang‐Dong Liu 1,✉
PMCID: PMC13441007  PMID: 40351112

Abstract

While host plants and endosymbionts have been implicated in influencing dietary breadth in polyphagous herbivores, the underlying mechanism remains vague. In this study, we focused on the food nutrition and nutrition provision of endosymbionts to elucidate the determination of dietary breadth in a polyphagous aphid Aphis gossypii. Our findings demonstrated that high sugar and riboflavin presence in food decreased aphid fitness, while Arsenophonus infections improved aphid performance. Aphids collected from cotton could not use cucumber whether they were infected with Arsenophonus or not, signifying a distinct specialization toward cotton. Further, both the Arsenophonus‐infected and free aphids fed on artificial diet varying in sugar titer failed to utilize cucumber. However, Arsenophonus‐free aphids attained the ability to utilize cucumber after feeding on the riboflavin‐free diet, but not on the riboflavin‐containing diet, indicating riboflavin and Arsenophonus‐absent expansion in dietary breadth. Notably, up‐regulated expression of riboflavin synthase genes of the obligated symbiont Buchnera aphidicola was detected in the Arsenophonus‐infected aphids which may provide more riboflavin. Arsenophonus promoting riboflavin synthesis in the obligated symbiont B. aphidicola and riboflavin ingestion enhancing host specialization of aphids to cotton modulate dietary breadth of A. gossypii.

Keywords: Aphis gossypii, Arsenophonus, Buchnera aphidicola, feeding experience, host specialization, riboflavin

Host plants and endosymbionts affect the ability of aphids to use some specific plants. Arsenophonus is a main facultative endosymbiont in cotton‐melon aphid, Aphis gossypii which exhibits obvious host specialization to cotton or cucumber. Riboflavin is an important nutrient for insects. Aphids collected from cotton cannot use cucumber, but they can when their endosymbionts Arsenophonus have been cured and fed on diet without riboflavin. Arsenophonus infections result in upregulation of riboflavin synthase genes of the obligated symbiont Buchnera in aphids. Arsenophonus‐mediated riboflavin is associated with the dietary breadth of this aphid.

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Introduction

Aphids constitute a significant group of sap‐sucking insect pests, posing threats to a wide range of agricultural crops and ornamental plants, leading to substantial crop losses (Blackman & Eastop, 2000; Sandhi & Reddy, 2020). Despite their polyphagous nature, many aphid populations demonstrate fidelity to a specific host plant, named host specialization (Dixon, 1998). For instance, Acyrtosiphon pisum, the pea aphid, shows host specialization to alfalfa or clover (Via, 1991), while Myzus persicae, the green peach aphid, exhibits a preference trade‐off between tobacco and pepper (Nikolakakis et al., 2003). Aphis gossypii, the cotton‐melon aphid, displays ubiquitous host specialization to cotton, cucumber, pepper, or chrysanthemum (Guldemond et al., 1994; Liu et al., 2003; Carletto et al., 2009). This prevalent host specialization within polyphagous aphid species underscores the flexibility of insect herbivores in adapting to diverse dietary breadth. Therefore, investigating the factor and mechanism influencing dietary breadth is crucial in polyphagous insects. Understanding these aspects offers profound insights into the evolution of insect dietary breadth, potentially yielding innovative strategies for insect pest management via manipulating crop composition and distribution.

Nutrient stress affects insect performance (Bruyn et al., 2002; Wetzel et al., 2016). Host plants play a pivotal role in providing essential nutrients for insect herbivores, and the quality of food is closely tied to a plant presence within dietary breadth of an insect. Aphids, as phloem sap feeders, exploit the rich sugar and dynamic content in plant phloem sap (Fisher, 1983; Hayashi & Chino, 1990; Winter et al., 1992; Ray & Savage, 2021; Broussard et al., 2023). Sugars play important roles in insects, acting as energy sources, osmolytes, and signaling molecules (Li et al., 2023; Chaudhari et al., 2025), which might influence aphid fitness and host range. The sugar concentration in phloem sap influences aphid performance and population fluctuation (Karley et al., 2002; Sadras et al., 2021). High sugar stress resulted in increased ingesting time of the pea aphid (Wang et al., 2021a). The number of adult aphids of Rophalosiphon padi decreased with increasing sugar content in wheat stems (Sadras et al., 2021). Sucrose affected the survival and diet consumption of aphids which was dependent on concentration in diet and aphid species; aphids M. persicae and Schizaphis graminum with a wide range of host plants were overall more adapted to feeding on a wider range of sucrose concentrations than the more host‐restricted aphid Aphis glycines (Puterka et al., 2017). Sucrose content varies across herbaceous and woody plants (Hayashi & Chino, 1986, 1990; Winter et al., 1992; Lohaus & Moellers, 2000), emphasizing that plants containing a specific sucrose titer meeting requirements of an aphid species fall within its host range.

Besides sugar, riboflavin, is another important nutrient for insects, particularly for piercing‐sucking mouthparts insects like rice leafhoppers Nephotettix cincticeps and mosquitoes Aedes aegypti (Matsumoto & Hattori, 2018; Wang et al., 2021b). Riboflavin functions as a coenzyme in flavoproteins facilitating enzymatic reactions required for the growth and survival of insects (Douglas, 2017). Riboflavin decay in the diet leads to the developmental arrest of Ae. aegypti larvae (Wang et al., 2021b), while riboflavin deficiency suppresses the fertility of symbiont‐cured whiteflies (Wang et al., 2020). However, insects lack the ability to synthesize riboflavin on their own (Douglas, 2017). Therefore, herbivorous insects may choose high riboflavin host plants, and riboflavin content may affect their dietary range.

Some specific experiences modify dietary breadth of insects (Fox & Savalli, 2000; Fox, 2006; Wu et al., 2013). Oviposition experiences on certain plants mediated host expansion of the seed beetle Stator limbatus (Fox & Savalli, 2000; Fox, 2006). Feeding experiences on zucchini, hibiscus, or cowpea altered the specialization of A. gossypii to cotton and cucumber (Liu et al., 2008; Wu et al., 2013). Moreover, feeding on a specific artificial diet induced the change in specialization of A. gossypii to cotton (Ma et al., 2019), demonstrating the effect of plant‐specific nutrients on dietary breadth.

Endosymbionts are intricately linked to aphid host specialization, with infections influencing changes in dietary breadth (Tsuchida et al., 2004, 2011; Ferrari et al., 2007; Wagner et al., 2015; Wulff & White, 2015). For instance, Regiella insecticola (PAUS) enhances pea aphid performance on white clover but not on alfalfa (Leonardo & Muiru, 2003), and Arsenophonus infections promote specialization of the cowpea aphid Aphis craccivora to locusts, altering host range of this polyphagous aphid (Wagner et al., 2015). Arsenophonus also facilitates rapid evolution in A. craccivora and this endosymbiont may have acted as a preadaptation of aphids to use a novel host plant (Lenhart & White, 2020). Host alternation in A. gossypii is associated with microbiome diversity (Ma et al., 2021). Relative abundance of Buchnera aphidicola significantly increases when cotton‐specialized aphids A. gossypii are transferred onto zucchini from cotton, and it presumes that this symbiont may break the specialization of aphids via providing nutrition (Xu et al., 2023). However, it is still obscure which nutrition provided by endosymbionts mediates dietary breadth of polyphagous insect herbivores.

Endosymbionts can provide specific nutrients for their hosts. Endosymbionts, such as B. aphidicola and Arsenophonus, in some insect species have the genetic capacity for synthesizing riboflavin (Nakabachi & Ishikawa, 1999; Shigenobu et al., 2000; Moriyama et al., 2015; Chong & Moran, 2018; Wang et al., 2021b). The obligate symbiont, B. aphidicola in pea aphids has genes encoding the biosynthetic pathway for riboflavin, and the aphids without Buchnera using rifampicin exhibited bad performance on the riboflavin‐free diet, compared to the aphids with Buchnera (Blow et al., 2020). The facultative symbiont, Arsenophonus, in aphids, whiteflies, and louse flies also possesses the complete pathway for riboflavin synthesis in genomes (Santos‐Garcia et al., 2018; Yorimoto et al., 2022; Rihova et al., 2023). Thus, endosymbionts may be associated with riboflavin provision to mediate dietary breadth or host specialization of polyphagous aphids.

Plant nutrients and symbionts collectively influence fitness and food niche width of insect populations (Wagner et al., 2015; Liu et al., 2024; Wang et al., 2024). While nutrient requirements crucially determine the dietary breadth of insect herbivores, symbionts can alter these requirements, leading to the expansion or contraction of host range (Wagner et al., 2015). Nevertheless, the specific plant nutrients and symbiont species influencing population fitness and dietary breadth of aphids are not fully understood. The cotton‐melon aphid A. gossypii, displaying strong specialization to cotton or cucumber plants (Liu et al., 2003; Wang et al., 2016) and harboring symbiotic bacteria, such as Buchnera and Arsenophonus (Xu et al., 2020; Chen et al., 2023), serves as an excellent model organism to explore the determinants of dietary breadth, particularly concerning plant nutrients and symbionts. This study hypothesizes that a specific nutrient, riboflavin, synthesized by symbionts, enhances host specialization and narrows the dietary breadth of cotton‐melon aphids. Feeding on diet containing riboflavin or infection with symbiont Arsenophonus restricts host shift of cotton‐melon aphids between cotton and cucumber which enhances host specialization (Fig. 1). Due to a great fluctuation in quantity of sugar and riboflavin in different plants and the riboflavin provision of symbionts, we examined the effects of sugar and riboflavin in diet on lifespan, reproduction, and host use of the Arsenophonus‐infected and cured aphids, and detected expression levels of riboflavin synthetase genes of Buchnera to indicate the ability of riboflavin provision. The results illustrated that riboflavin nutrient and Arsenophonus symbionts modulate dietary breadth of a polyphagous aphid.

Fig. 1.

Fig. 1

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A presume mode of the host specialization of aphids mediated by symbionts and food nutrient. A black cross (×) presents failure to host shift, and a green check mark (√) presents success of host shift. + and − marks mean the presence and deficiency of a nutrient in an aphid diet, respectively.

Materials and methods

Aphids and plants

Three distinct genotypes of A. gossypii, denoted as CA2, CA3, and CA4, were utilized in this study (Tian et al., 2019; Chang et al., 2022). These aphids, originally collected from cotton and subsequently reared in the laboratory on cotton seedlings, were infected solely with the facultative endosymbiont Arsenophonus (Ars+ strain). Additionally, Arsenophonus‐cured strains (Ars‐) of CA2, CA3, and CA4 were established in the laboratory using antibiotics 4 years ago (Tian et al., 2019; Chang et al., 2022). These 3 genotypes of aphids were unable to utilize cucumber, earning them the designation of the cotton‐origin or cotton‐specialized aphids. Cotton and cucumber seedlings were cultivated in a controlled warm greenhouse environment. All aphid strains were reared using cotton seedlings in a growth chamber at 25 ± 1 °C, 70% ± 10% relative humidity and 16 h L: 8 h D. Due to the possibility of losing or regaining symbiotic bacteria during the rearing process, diagnostic polymerase chain reaction (PCR) was performed to determine presence or absence of Arsenophonus in an aphid using the specific primer based on fbaA gene of Arsenophonus (Chang et al., 2022), and 10 aphids were randomly collected from tested samples and examined. If 100% aphids from infected strains were still infected with Arsenophonus or 100% aphids from cured strains were uninfected, those remaining samples were used as Arsenophonus‐infected or cured aphids in the following experiments.

Sugar and riboflavin‐controlled diet

The dietary sucrose titer crucial for the growth, survival, and reproduction of A. gossypii was determined to be optimal within the range from 20% to 30% (Auclair, 1967). Two distinct artificial diets were formulated for aphid rearing. The artificial diets were mainly composed of amino acids, vitamins, and minerals. The details on the composition are listed in Tables S1 and S2. One diet was controlled sucrose titer at 150 g/L (15%) or 300 g/L (30%) without riboflavin (Table S1), while the other was controlled riboflavin at 0 and 25 mg/L, containing 30% sucrose (Table S2). Aphids were fed using the double membrane sandwich method, and aphid stylets punctured the Parafilm for feeding (Ma et al., 2019).

Establishment of life tables of aphid populations

Life table methodology was employed to investigate the influence of Arsenophonus infections and dietary nutrients on aphid survival and reproduction. Life tables were constructed for Ars+ and Ars‐ strains of CA2, CA3, and CA4 on 4 distinct diets, named sucrose‐controlled (150 and 300 g/L) and riboflavin‐controlled (0 and 25 mg/L) diet. Each strain was replicated 5 times on each diet. Ten newly born aphids were introduced onto the specified artificial diet as the starting population, and their survival and reproduction were monitored daily until the demise of the entire cohort. Net reproductive rate (R 0) and average generational lifespan (T) were computed to assess the reproductive and survival capabilities of aphid populations on a specific diet, respectively (Liu et al., 2008).

Measurement of soluble sugar contents in cotton and cucumber leaves

The cotton and cucumber seedlings with 4 true leaves planted in a controlled warm greenhouse at 25 °C were used to examine contents of soluble sugar. The cotton and cucumber were conventional varieties Lufeng No1 and Lanlaohan, respectively. Two cucumber leaves (2.38−2.64 g) or 4 cotton leaves (1.94−2.13 g) were collected as a sample and then dried at 55 °C and ground into fine powder. One gram of powder from a sample was used to examine the content of soluble sugar content using the anthrone‐H2SO4 colorimetry method (Chang et al., 2022). Three replicates were performed for both cotton and cucumber leaves.

Host shift of aphids

The population dynamics of the Ars+ and Ars‐ strains of the CA2, CA3, and CA4 on cotton and cucumber leaves were examined to explore whether Arsenophonus infections would impact the utilization of aphids to cotton and cucumber. Thirty 5‐d‐old aphids were transferred onto a leaf in a Petri dish, and then the number of aphids was surveyed every 2 d till all aphids died or the number exceeded 400. Five replicates were performed for each strain on cucumber leaves, and 3−5 replicates were performed for each strain on cotton leaves. The petioles of cotton and cucumber were wrapped in wet cotton wool to keep leaves fresh.

To examine the impact of feeding experiences on sucrose and riboflavin‐controlled diet on host specialization of aphids, host shift from cotton onto cucumber leaves was executed. Thirty aphids from Ars+ and Ars‐ strains of CA2, CA3, and CA4 on cotton were reared on nutrient‐controlled diets for 5 d, and then transferred onto cucumber leaves in Petri dishes. Population dynamics were monitored, and the number of aphids, including transferred individuals and their offspring, was recorded every 3 d. The survey duration after the host shift extended to 27 d, as aphids directly transferred onto cucumber leaves could survive for 21 or 27 d (Liu et al., 2008; Ma et al., 2019). Each strain reared on each nutrient‐controlled diet or cotton leaves was replicated 10 times.

Examination of gene expression level of riboflavin synthase of Buchnera

The transcriptomes of B. aphidicola, the obligate symbiont, in the Ars+ and Ars‐ strains of 2 genotypes CA2 and CA4 aphids were sequenced using Illumina HiSeq™ by the Novogene Experimental Department, Beijing Novogene Bioinformatics Technology Co. Ltd, and the CA3 was used for quantitative PCR (qPCR) test. Three hundred aphids (5‐d‐old) CA2 or CA4 reared on cotton were collected for total RNA extraction. The Ribo‐zero rRNA Removal Kit was used to enrich messenger RNA (mRNA) of Buchnera. Twelve complementary DNA (cDNA) libraries with 3 biological replicates for each strain and genotype were constructed and sequenced on HiSeq platform to obtain raw reads. Clean reads removed the adapter and low‐quality sequences were assembled and annotated based on the genome of B. aphidicola (GenBank, NZ_CP056771.1). Gene expression levels were analyzed using HTSeq (v0.6.1) to compute the FPKM (expected number of fragments per kilobase of transcript sequence per millions base pairs sequenced). Genes in the riboflavin biosynthetic pathway of Buchnera, ribA, ribB, ribD1/ribD2, ribE, and ribH were identified (Fig. S1) in the transcriptomes of Ars+ and Ars‐ strains of CA2 and CA4 aphids, and their expression levels were assessed using the FPKM values in the transcriptome data.

The relative expression levels of 6 genes of Buchnera, ribA, ribB, ribD, ribE, ribF, and ribH in the Arsenophonus‐infected and free strains of CA3 were examined using qPCR method based on the primers listed in Table S3. The actin and EF1‐α of aphids were used as reference genes (Yang et al., 2014; Ma et al., 2016; Zhang et al., 2016). Fifty 5‐d‐old Arsenophonus‐infected and free aphids on cotton were collected as a sample to extract total RNA using Trizol reagent (TaKaRa, Japan). The cDNA was synthesized using TaKaRa PrimeScriptTM RT Master Mix (Perfect Real Time) Kit. Two microliters of cDNA, 10 µL 2× TB Green Premix Ex Taq II (Tli RNaseH Plus), 0.4 µL forward and reverse primers (10 µmol/L), 0.4 µL ROX reference Dye II, and 6.8 µL ddH2O were used as the qPCR reaction. The qPCR was performed in Applied Biosystems QuantStudio™ 6 (Carlsbad, CA) with 95 °C for 30 s, and then 40 cycles with 95 °C for 5 s and 60 °C for 30 s. Three biological replications were performed. The relative expression level of a gene was computed using the 2−ΔΔCt method.

Data analysis

The effects of Arsenophonus infections and sucrose or riboflavin titers in the diet on R 0 and T of CA2, CA3, CA4 aphid populations were subjected to two‐way analysis of variance in the general linear model (GML). The effects of Arsenophonus and aphid genotype on gene expression level of riboflavin synthase of Buchnera were also analyzed using GML. Differences in R 0 or T between Ars+ and Ars‐ strains, as well as in soluble sugar contents between cotton and cucumber leaves, were analyzed using Student's t‐test. The effects of Arsenophonus infection and aphid genotype on the population sizes of aphids on cucumber at different days were tested using GML with repeated measurements. Effects of Arsenophonus infection and sucrose or riboflavin titers in the diet on population dynamics of aphids on cucumber leaves were also analyzed using the GML model, treating population sizes surveyed at different dates as repeated measurements. The effects of Arsenophonus infections and aphid genotypes on FPKM values were analyzed by GLM, and these FPKM values were normalized using z‐score method and the heatmap was made in GraphPad Prism 7.0. The relative expression levels of 6 riboflavin genes were compared between the Arsenophonus‐infected and uninfected aphids using Student's t‐test. All the statistics analyses were conducted using IBM SPSS Statistics V19.

Results

Arsenophonus infections and sugar titer influence aphid performance

Arsenophonus infections (F 1, 48 = 102.387, P < 0.001) and dietary sugar titer (F 1, 48 = 72.271, P < 0.001) significantly impacted aphid performance. Arsenophonus infections boosted reproduction (net reproductive rate, R 0) of the aphid populations while high sugar titer overall suppressed them (Fig. 2A, B, C). Aphid genotype also affected the reproduction of aphids by interacting with sugar titer (F 2, 48 = 4.741, P = 0.013) or Arsenophonus infection (F 2, 48 = 4.233, P = 0.020). The net reproductive rates of Arsenophonus‐infected and cured populations on 15% sugar diet were overall higher than that on 30% sugar diet, suggesting a high sugar stress on aphids. There was an exception that the net reproductive rate of the Arsenophonus‐infected CA4 on the 15% sugar diet was similar to that on the 30% diet (t = 1.135, df = 8, P = 0.289) and exhibited a large variation (Fig. 2C). The sugar titer (F 1, 48 = 11.979, P = 0.0011) and interaction between aphid genotype and Arsenophonus infection (F 2, 48 = 8.028, P = 0.001) significantly affected survival (average generational lifespan) of aphid populations (Fig. 2D, E, F). Notably, Arsenophonus infections significantly increased survival of CA3 on the 30% sugar diet (t = 3.214, df = 8, P = 0.012; Fig. 2E), but no effects with the 15% diet and the other 2 genotype aphids on both diets (Fig. 2D, E, F).

Fig. 2.

Fig. 2

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Arsenophonus infections and sugar titer affect the reproduction and survival of aphid populations. The net reproductive rate R 0 (A, B, C) and average generational lifespan T (D, E, F) of Arsenophonus‐infected (Ars+) and cured (Ars‐) aphid populations CA2, CA3, CA4 reared on 15% and 30% sugar diet. The P‐values indicate the significant levels of differences between Arsenophonus‐infected (Ars+) and cured (Ars‐) strains based on Student's t‐test.

Arsenophonus infections and riboflavin in diet affect aphid performance

On the riboflavin‐free diet, Arsenophonus infections did not affect the net reproductive rate of CA2 and CA4 aphids, while it significantly increased the net reproductive rate of CA3 aphids (t = 10.375, df = 8, P = 0.00001; Fig. 3A), suggesting an aphid genotype‐dependent effect. Arsenophonus infections did not affect the average generational lifespan of aphids on the riboflavin‐free diet (Fig. 3B). Additionally, the performance of CA3 on the riboflavin‐containing and free diets was compared. The results showed that riboflavin presence in diet significantly decreased the net reproductive rate (Ars+ strain: t = 6.715, df = 8, P = 0.00015; Ars‐ strain: t = 5.982, df = 8, P = 0.00033) and average generational lifespan (Ars+ strain: t = 9.995, df = 8, P = 0.00001; Ars‐ strain: t = 2.745, df = 8, P = 0.025) of the Arsenophonus‐infected and cured aphids (Fig. S2), indicating the riboflavin stress on aphids.

Fig. 3.

Fig. 3

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Arsenophonus infections affect reproduction and survival of aphid populations feeding on the riboflavin‐free diet. The net reproductive rate (A) and average generational lifespan (B) of CA2, CA3, and CA4 genotypes. The P‐values indicate the significant levels of differences between Arsenophonus‐infected (Ars+) and cured (Ars‐) strains based on Student's t‐test.

Plant nutrients and host specialization of aphids

The soluble sugar content in cotton leaves was significantly lower than that in cucumber leaves (t = 12.46, df = 4, P = 0.0002; Fig. 4A). Both the Arsenophonus‐infected and cured CA2, CA3, and CA4 aphid populations boosted on cotton, but they failed to establish populations on cucumber, surviving only 8−32 d and displaying an inability to utilize cucumber as a host plant (Fig. 4B, C, D).

Fig. 4.

Fig. 4

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Cotton‐origin aphids cannot use cucumber. Soluble sugar content in cotton and cucumber leaves (A) and population dynamics of cotton‐origin aphids infected with (Ars+) and cured Arsenophonus (Ars‐) CA2 (B), CA3 (C), and CA4 (D) on cotton and cucumber leaves. ***P < 0.0001.

Effects of Arsenophonus infections and sugar titer on dietary breadth

Arsenophonus infections suppressed population development of cotton‐origin CA2 on cucumber (F 1, 36 = 12.748, P = 0.001; Fig. 5A), while the effects on CA3 (F 1, 36 = 1.741, P = 0.195; Fig. 5B) and CA4 (F 1, 36 = 0.755, P = 0.391; Fig. 5C) were not significant. Feeding experience on diet with 15% and 30% sugar did not significantly affect population sizes of CA2 (F 1, 36 = 1.789, P = 0.189; Fig. 5A), CA3 (F 1, 36 = 0.345, P = 0.560; Fig. 5B), and CA4 (F 1, 36 = 0.007, P = 0.934; Fig. 5C) on cucumber, and these cotton‐origin CA2 and CA3 aphids still could not establish populations on cucumber after feeding experience on sugar‐controlled diet, although 1 out of 10 replicates for the CA4 Arsenophonus‐cured strain maintained a high density on cucumber after being reared on the 30% sugar diet for 5 d (Fig. 5C), suggesting that the high sugar stress might be a potential component for inducing change in dietary breadth of aphids.

Fig. 5.

Fig. 5

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Effects of Arsenophonus infections and sugar titer in diet on dietary breadth in aphids. The population sizes of cotton‐origin CA2 (A), CA3 (B), and CA4 (C) aphids on cucumber leaves after feeding on 15% and 30% sugar diet for 5 d.

Effects of Arsenophonus infections and riboflavin on dietary breadth

Arsenophonus infections and riboflavin in diet significantly affected population sizes of cotton‐origin CA2 (Arsenophonus infection: F 1, 36 = 14.858, P = 0.0005; riboflavin: F 1, 36 = 28.856, P < 0.0001; Fig. 6A), CA3 (Arsenophonus infection: F 1, 36 = 17.854, P = 0.0002; riboflavin: F 1, 36 = 11.659, P = 0.002; Fig. 6B), and CA4 (Arsenophonus infection: F 1, 36 = 5.071, P = 0.031; riboflavin: F 1, 36 = 28.453, P < 0.0001; Fig. 6C) aphids on cucumber. The population size of Arsenophonus‐cured aphids with feeding experience on riboflavin‐free diet was significantly larger than that of the Arsenophonus‐infected aphids. Arsenophonus‐cured aphids fed on the riboflavin‐free diet for 5 d gained the ability to use cucumber, indicating an expansion in dietary breadth (Fig. 6).

Fig. 6.

Fig. 6

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Absence of Arsenophonus in aphids and riboflavin in diet expands dietary breadth in aphids. Population sizes of Arsenophonus‐infected (Ars+) and cured (Ars‐) CA2 (A), CA3 (B), and CA4 (C) aphids on cucumber leaves after feeding on riboflavin‐present and free diet for 5 d.

Arsenophonus promotes expression of riboflavin synthase genes of Buchnera

Arsenophonus infections promoted the expression of riboflavin synthase genes of the obligated symbiont Buchnera (Fig. 7). The expression levels of 6 genes of riboflavin synthetase of Buchnera in the Ars+ and Ars‐ strains differed based on the transcriptome. Arsenophonus infections significantly increased expression levels of ribD2 (F 1, 8 = 7.983, P = 0.022), ribE (F 1, 8 = 6.166, P = 0.038), and ribH (F 1, 8 = 15.602, P = 0.004), and the effect of Arsenophonus infections on expression level of ribB was dependent on aphid genotype (F 1, 8 = 12.054, P = 0.008), while the expression levels of ribA (F 1, 8 = 1.448, P = 0.263) and ribD1 (F 1, 8 = 3.560, P = 0.096) were not impacted by Arsenophonus infections. Expression levels of all 6 genes were not affected by aphid genotype (ribA: F 1, 8 = 0.962, P = 0.355; ribB: F 1, 8 = 0.271, P = 0.617; ribD1: F 1, 8 = 2.228, P = 0.174; ribD2: F 1, 8 = 1.181, P = 0.309; ribE: F 1, 8 = 0.156, P = 0.703; ribH: F 1, 8 = 1.656, P = 0.234; Fig. 7A).

Fig. 7.

Fig. 7

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Arsenophonus infections promote the expression of Buchnera genes in the riboflavin biosynthetic pathway. Heatmap of the expression levels of riboflavin synthase genes of Buchnera in Arsenophonus‐infected (Ars+) and cured (Ars‐) aphids (CA2 and CA4 genotypes) based on the z‐score of fragments per kilobase of transcript sequence per millions base pairs in transcriptome analyses (A), biosynthesis pathway of riboflavin in bacteria (B) (Garcia‐Angulo, 2017; Santos‐Garcia et al., 2018; Blow et al., 2020), and the relative expression levels of 6 genes in the biosynthetic pathway of riboflavin in Buchnera examined by quantitative polymerase chain reaction (C). * and ** mean significant difference between Arsenophonus‐infected (Ars+) and free (Ars‐) strains at P = 0.05 and 0.01 levels, respectively, and ns means no significant difference at P = 0.05 level by Student's t‐test. GTP: guanosin‐5ʹ‐triphosphate. B2: riboflavin. R5P, ribulose‐5‐phosphate.

Six genes, ribA, ribB, ribD, ribE, ribF, and ribH, need to be involved in the riboflavin biosynthetic pathway in Buchnera (Fig. 7B). Based on qPCR detection, the relative expression levels of ribA (t = 3.972, df = 4, P = 0.017), ribB (t = 7.943, df = 4, P = 0.001), ribE (t = 7.114, df = 4, P = 0.002), ribF (t = 4.527, df = 4, P = 0.011), and ribH (t = 7.074, df = 4, P = 0.002) of Buchnera were significantly higher in the Arsenophonus‐infected aphids than that in the Arsenophonus‐free aphids, while the expression level of ribD was not significantly different between them (t = 2.372, df = 4, P = 0.077; Fig. 7C).

Discussion

Higher sucrose titer in the diet adversely impacted performance of cotton‐specialized aphids, leading to decreased reproduction regardless of Arsenophonus infections. High‐titer sugar induces nutrient stress imposed on aphids, aligning with findings in the pea aphid (Wilkinson et al., 1997; Douglas, 2006). Sugar‐rich phloem sap obliges aphids to cope with osmotic stress, potentially causing reduced feeding and, consequently, poor aphid performance (Douglas, 2006; Dinant et al., 2010; Nalam et al., 2021). Although aphids have adapted to sugar‐rich food, the higher titer of sugar in diet becomes a nutrient stress. It has been found that external stress may alter insect preference to novel host plants (Xu et al., 2014; Fukano et al., 2016; Savkovic et al., 2016; Liu et al., 2017). The seed beetle, Acanthoscelides obtectus, adapted well to a less preferable plant host, chickpea, Cicer arietinum, by evolving a life‐history strategy when enforced to live on this plant for 48 generations from the ancestral host common bean (Savkovic et al., 2016). The ragweed leaf beetles Ophraella communa did not use Ambrosia trifida as a host plant in their native North America, but they extensively utilized it when introduced to Japan, meaning host plant range expansion of the introduced O. communa (Fukano et al., 2016). Migratory North American monarchs Danaus plexippus had higher performance than derived nonmigratory populations on all host plants (Freedman et al., 2019). Two generations of feeding experience of A. gossypii collected from hibiscus on the cucumber leaves infected with a phytopathogen, Pseudoperonospora cubensis, attained the ability to use intact cucumber plants (Ali et al., 2021). Feeding experience on specific hosts or diet alter the host range or dietary breadth of cotton‐specialized aphids (Wu et al., 2013; Liu et al., 2017; Ma et al., 2019). In this study, feeding experience under high sugar stress (30%) induced the improvement in performance of Arsenophonus‐free aphids on cucumber leaves, but this improvement was significant only in a small part of CA4 aphids. This result implies that dietary sugar stress might be a potential component for inducing the expansion of dietary breadth of aphids, and CA4 might incur population differentiation in response to sugar stress, exhibiting the large variation in reproduction under high sugar stress (Fig. 2C).

While riboflavin is synthesized by plants (Barrera & Picha, 2014; Nasreen et al., 2022), its presence in the diet did not significantly affect survival and growth of pea aphids, irrespective of Buchnera presence (Blow et al., 2020). However, omission of riboflavin in the diet significantly improved survival, growth, and reproduction of pea aphids (Boisvert & Auclair, 1981). Individual deletion of riboflavin in diet did not result in changes of nymphal pea aphids in survival and relative growth rates, compared to aphids on the complete diet with all B vitamins (Blow et al., 2020), indicating that riboflavin is not required. We found that riboflavin in diet was a nutrient stress for cotton‐specialized A. gossypii. However, riboflavin stress did not induce the change in host specialization of A. gossypii. On the contrary, removal of riboflavin from diet expanded the host range of the Arsenophonus‐free aphids which attained the ability to use cucumber after feeding on the riboflavin‐free diet, but not the Arsenophonus‐infected aphids. These results indicate that dietary riboflavin enhances host specialization of aphids to cotton.

Given that phytochemicals affect host range expansion of insects via promoting the maintenance of beneficial interactions between insects and microbial symbionts (Erbilgin, 2019), microbes play a crucial role in mediating the shift of host plants in phytophagous insects (Adams et al., 2013; Vilanova et al., 2016; Lange et al., 2023). Endosymbiont infections, such as Arsenophonus, enhance aphid specialization to specific host plants (Leonardo & Muiru, 2003; Tsuchida et al., 2004; Wagner et al., 2015). In this study, we found that the Arsenophonus‐infected strain of aphids performed poorly and perished on cucumber leaves. But the Arsenophonus‐free strain was relatively better and established populations on cucumber leaves after feeding on the riboflavin‐free diet. The role of symbiont in determining dietary breadth of aphids was linked to nutrients in plants (Fig. 1). The fact that aphid populations on different host plants exhibited different infection frequencies of endosymbiont species and specialization (Vorburger et al., 2017; Tian et al., 2019) and these inconsistent results on a symbiont affecting host use of an aphid species may attribute to the role of host plant nutrient and the specific feeding experience of aphids, besides the genotype of aphid and symbiont (Leonardo, 2004; Tsuchida et al., 2004; Ferrari et al., 2007). The intricate relationship between aphid, symbiont, and plant, influenced by plant nutrient composition, contributes to evolution of dietary breadth in herbivorous insects.

The obligate symbiont Buchnera is essential to A. gossypii. The facultative symbiont Arsenophonus improves growth performance of A. gossypii under amino acid stress synergistically with Buchnera (Tian et al., 2023). Buchnera possesses synthase genes of B vitamin and provides aphids with riboflavin (Nakabachi & Ishikawa, 1999; Blow et al., 2020). In the eusocial aphid Ceratovacuna japonica, Buchnera lost all genes responsible for riboflavin biosynthesis, but these genes were retained in the Arsenophonus genome, suggesting co‐obligate symbionts (Yorimoto et al., 2022). In this study, we identified riboflavin synthase genes of Buchnera and found that Arsenophonus promoted their expression levels which might provide more riboflavin for aphids. Aphids fed on diet with riboflavin performed poorer on cucumber leaves. Arsenophonus‐induced host specialization of aphids may attribute to the enhancement of riboflavin provision in another endosymbiont Buchnera. Nutritional deficiency of plants restricts insect utilization as a host. Symbionts can synthetize the deficient nutrition, such as amino acids and vitamins (Yorimoto et al., 2022; Rihova et al., 2023). If insects carry a symbiont which can provide the deficient nutrition in a specific plant, they likely expand their host range to this plant. Therefore, the interaction between Arsenophonus and Buchnera to regulate the dietary breadth of A. gossypii found in this study may influence the long‐term evolution of host specialization in aphids.

Given host specialization enhancement induced by riboflavin in diet or Arsenophonus infections, aphid population expansion toward other crops may be suppressed via symbiont infections or increasing riboflavin content in crops. So, altering nutritional composition of crops and symbiont infections of insects may be a potential strategy for pest management, especially in the mixed planting areas of different crops.

In this study, we only examined the effects of food nutrition and endosymbionts on dietary breadth in the species A. gossypii, and whether these findings are likely to be generalizable to other aphid species or polyphagous herbivores is worth studying. Arsenophonus genomes preserve all genes related to riboflavin synthesis (Santos‐Garcia et al., 2018; Yorimoto et al., 2022; Rihova et al., 2023). Besides promoting riboflavin synthesis in Buchnera, whether Arsenophonus bacteria themselves can synthesize riboflavin in A. gossypii is still unknown. The mechanisms by which Arsenophonus regulates the gene expression of Buchnera and interacts with food to affect dietary changes of aphids also need to be further investigated.

In sum, this study identifies that riboflavin in food and facultative symbiont Arsenophonus enhance host specialization of aphids. While high titer of sugar or riboflavin presence suppresses fitness of aphid populations, riboflavin deficiency induces the expansion in dietary breadth of Arsenophonus‐free aphids, but not Arsenophonus‐infected aphids. Arsenophonus leads to upregulation of synthase genes of riboflavin in Buchnera. As a result, Arsenophonus‐enhanced host specialization of aphids may be associated with riboflavin provision. Further investigations into the synergistic mechanism of Arsenophonus and Buchnera in promoting change in dietary breadth of aphids are warranted.

Disclosure

All authors have seen and agree with the contents of the manuscript and there is no conflict of interest, including specific financial interest and relationships and affiliations relevant to the subject of the manuscript.

Supporting information

Fig. S1 The sequences of 6 riboflavin genes of Buchnera.

INS-33-1567-s001.docx (109.5KB, docx)

Fig. S2 Net reproductive rate and average generational lifespan of CA3 populations infected with Arsenophonus (Ars+) or not (Ars‐) on the diet with riboflavin or not.

INS-33-1567-s001.docx (109.5KB, docx)

Table S1 Nutritional composition of the artificial diet with 15% sucrose.

INS-33-1567-s001.docx (109.5KB, docx)

Table S2 Nutritional composition of the artificial diet with 25 mg/L riboflavin.

INS-33-1567-s001.docx (109.5KB, docx)

Table S3 Primers used in this study.

INS-33-1567-s001.docx (109.5KB, docx)

Acknowledgments

This study was supported by the Jiangsu Agricultural Science and Technology Innovation Fund (CX23[1037]) and National Natural Science Foundation of China (31672034).

Contributor Information

Hui‐Fang Guo, Email: guohfjaas@163.com.

Xiang‐Dong Liu, Email: liuxd@njau.edu.cn.

References

  1. Adams, A.S. , Aylward, F.O. , Adams, S.M. , Erbilgin, N. , Aukema, B.H. , Currie, C.R. et al. (2013) Mountain pine beetles colonizing historical and naive host trees are associated with a bacterial community highly enriched in genes contributing to terpene metabolism. Applied and Environmental Microbiology, 79, 3468–3475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Ali, F. , Hu, X. , Wang, D. , Yang, F. , Guo, H. and Wang, Y. (2021) Plant pathogen‐mediated rapid acclimation of a host‐specialized aphid to a non‐host plant. Ecology and Evolution, 11, 15261–15272. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Auclair, J.L. (1967) Effects of pH and sucrose on rearing the cotton aphid, Aphis gossypii, on a germ‐free and holidic diet. Journal of Insect Physiology, 13, 431–446. [Google Scholar]
  4. Barrera, W.A. and Picha, D.H. (2014) Ascorbic acid, thiamin, riboflavin, and vitamin B6 contents vary between sweetpotato tissue types. Hortscience, 49, 1470–1475. [Google Scholar]
  5. Blackman, R.L. and Eastop, V.F. (2000) Aphids on the World's Crops: an Identification and Information Guide. Wiley. [Google Scholar]
  6. Blow, F. , Bueno, E. , Clark, N. , Zhu, D.T. , Chung, S.H. , Güllert, S. et al. (2020) B‐vitamin nutrition in the pea aphid‐Buchnera symbiosis. Journal of Insect Physiology, 126, 104092. [DOI] [PubMed] [Google Scholar]
  7. Boisvert, J.M. and Auclair, J.L. (1981) Influence de la riboflavin sur les besoins globaux en vitamins hydrosolubles chez le puceron du pois, Acyrthosiphon pisum . Canadian Journal of Zoology, 59, 164–169. [Google Scholar]
  8. Broussard, L. , Abadie, C. , Lalande, J. , Limami, A.M. , Lothier, J. and Tcherkez, G. (2023) Phloem sap composition: what have learnt from metabolomics? International Journal of Molecular Sciences, 24, 6917. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Bruyn, L.D. , Scheirs, J. and Verhagen, R. (2002) Nutrient stress, host plant quality and herbivore performance of a leaf‐mining fly on grass. Oecologia, 130, 594–599. [DOI] [PubMed] [Google Scholar]
  10. Carletto, J. , Lombaert, E. , Chavigny, P. , Brévault, T. , Lapchin, L. and Vanlerberghe‐Masutti, F. (2009) Ecological specialization of the aphid Aphis gossypii Glover on cultivated host plants. Molecular Ecology, 18, 2198–2212. [DOI] [PubMed] [Google Scholar]
  11. Chang, C.Y. , Sun, X.W. , Tian, P.P. , Miao, N.H. , Zhang, Y.L. and Liu, X.D. (2022) Plant secondary metabolite and temperature determine the prevalence of Arsenophonus endosymbionts in aphid populations. Environmental Microbiology, 24, 3764–3776. [DOI] [PubMed] [Google Scholar]
  12. Chaudhari, B.Y. , Pradhan, A.G. and Joshi, R.S. (2025) Metabolic gatekeepers: dynamic roles of sugar transporters in insect metabolism and physiology. Insect Molecular Biology, 34, 1–18. [DOI] [PubMed] [Google Scholar]
  13. Chen, R. , Luo, J. , Zhu, X. , Wang, L. , Zhang, K. , Li, D. et al. (2023) Dynamic changes in species richness and community diversity of symbiotic bacteria in five reproductive morphs of cotton aphid Aphis gossypii Glover (Hemiptera: Aphididae). Frontiers in Microbiology, 13, 1086728. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Chong, R.A. and Moran, N.A. (2018) Evolutionary loss and replacement of Buchnera, the obligate endosymbiont of aphids. The ISME Journal, 12, 898–908. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Dinant, S. , Bonnemain, J.L. , Girousse, C. and Kehr, J. (2010) Phloem sap intricacy and interplay with aphid feeding. Comptes Rendus Biologies, 333, 504–515. [DOI] [PubMed] [Google Scholar]
  16. Dixon, A.F.G. (1998) Aphid Ecology (2nd edition). Chapman & Hall. [Google Scholar]
  17. Douglas, A.E. (2006) Phloem‐sap feeding by animals: problems and solutions. Journal of Experimental Botany, 57, 747–754. [DOI] [PubMed] [Google Scholar]
  18. Douglas, A.E. (2017) The B vitamin nutrition for insects: the contributions of diet, microbiome and horizontally acquired genes. Current Opinion in Insect Science, 23, 65–69. [DOI] [PubMed] [Google Scholar]
  19. Erbilgin, N. (2019) Phytochemicals as mediators for host range expansion of a native invasive forest insect herbivore. New Phytologist, 221, 1268–1278. [DOI] [PubMed] [Google Scholar]
  20. Ferrari, J. , Scarborough, C.L. and Godfray, H.C.J. (2007) Genetic variation in the effect of a facultative symbiont on host‐plant use by pea aphids. Oecologia, 153, 323–329. [DOI] [PubMed] [Google Scholar]
  21. Fisher, D.B. (1983) Year‐round collection of willow sieve‐tube exudate. Planta, 159, 529–533. [DOI] [PubMed] [Google Scholar]
  22. Fox, C.W. (2006) Colonization of a new host by a seed‐feeding beetle: genetic variation, maternal experience, and the effect of an alternate host. Annales Zoologici Fennici, 43, 239–247. [Google Scholar]
  23. Fox, C.W. and Savalli, U.M. (2000) Maternal effects mediate host expansion in a seed‐feeding beetle. Ecology, 81, 3–7. [Google Scholar]
  24. Freedman, M.G. , Jason, C. , Ramirez, S.R. and Strauss, S.Y. (2019) Host plant adaptation during contemporary range expansion in the monarch butterfly. Evolution; International Journal of Organic Evolution, 74, 377–391. [DOI] [PubMed] [Google Scholar]
  25. Fukano, Y. , Doi, H. , Thomas, C.E. , Takata, M. , Koyama, S. and Satoh, T. (2016) Contemporary evolution of host plant range expansion in an introduced herbivorous beetle Ophraella communa . Journal of Evolutionary Biology, 29, 757–765. [DOI] [PubMed] [Google Scholar]
  26. Garcia‐Angulo, V.A. (2017) Overlapping riboflavin supply pathways in bacteria. Critical Reviews in Microbiology, 43, 196–209. [DOI] [PubMed] [Google Scholar]
  27. Guldemond, J.A. , Tigges, W.T. and De Vrijer, W.F. (1994) Host races of Aphis gossypii (Homotera: Aphididae) on cucumber and chrysanthemum. Environmental Entomology, 23, 1235–1240. [Google Scholar]
  28. Hayashi, H. and Chino, M. (1986) Collection of pure phloem sap from wheat and its chemical composition. Plant Cell Physiology, 27, 1387–1393. [Google Scholar]
  29. Hayashi, H. and Chino, M. (1990) Chemical composition of phloem sap from the uppermost internode of the rice plant. Plant Cell Physiology, 31, 247–251. [Google Scholar]
  30. Karley, A.J. , Douglas, A.E. and Parker, W.E. (2002) Amino acid composition and nutritional quality of potato leaf phloem sap for aphids. Journal of Experimental Biology, 205, 3009–3018. [DOI] [PubMed] [Google Scholar]
  31. Lange, C. , Boyer, S. , Bezemer, T.M. , Lefort, M.C. , Dhami, M.K. , Biggs, E. et al. (2023) Impact of intraspecific variation in insect microbiomes on host phenotype and evolution. The ISME Journal, 17, 1798–1807. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Lenhart, P.A. and White, J.A. (2020) Endosymbionts facilitate rapid evolution in a polyphagous herbivore. Journal of Evolutionary Biology, 33, 1507–1511. [DOI] [PubMed] [Google Scholar]
  33. Leonardo, T.E. and Muiru, G.T. (2003) Facultative symbionts are associated with host plant specialization in pea aphid populations. Proceedings of the Royal Society B, 270, S209–S212. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Leonardo, T.E. (2004) Removal of a specialization‐associated symbiont does not affect aphid fitness. Ecology Letters, 7, 461–468. [Google Scholar]
  35. Li, X. , Zhou, Y. and Wu, K. (2023) Biological characteristics and energy metabolism of migrating insects. Metabolites, 13, 439. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Liu, S. , Liu, X.B. , Zhang, T.T. , Bai, S.X. , He, K.L. , Zhang, Y.J. et al. (2024) Effects of host plants on aphid feeding behavior, fitness, and Buchnera aphidicola titer. Insect Science, 10.1111/1744-7917.13428. [DOI] [PubMed] [Google Scholar]
  37. Liu, X.D. , Xu, T.T. and Lei, H.X. (2017) Refuges and host shift pathways of host‐specialized aphids Aphis gossypii . Scientific Reports, 7, 2008. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Liu, X.D. , Zhai, B.P. and Zhang, X.X. (2003) Studies on the host biotypes and its cause of cotton aphid in Nanjing, China. Scientia Agricultura Sinica, 36, 54–58. [Google Scholar]
  39. Liu, X.D. , Zhai, B.P. and Zhang, X.X. (2008) Specialized host‐plant performance of the cotton aphid is altered by experience. Ecological Research, 23, 919–925. [Google Scholar]
  40. Lohaus, G. and Moellers, C. (2000) Phloem transport of amino acids in two Brassica napus L. genotypes and one B. carinata genotype in relation to their seed protein content. Planta, 211, 833–840. [DOI] [PubMed] [Google Scholar]
  41. Ma, K.S. , Li, F. , Liang, P.Z. , Che, X.W. , Liu, Y. and Gao, X.W. (2016) Identification and validation of reference genes for the normalization of gene expression data in qRT‐PCR analysis in Aphis gossypii (Hemiptera: Aphididae). Journal of Insect Science, 16, 17. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Ma, L. , Li, M.Y. , Chang, C.Y. , Chen, F.F. , Hu, Y. and Liu, X.D. (2019) The host range of Aphis gossypii is dependent on aphid genetic background and feeding experience. PeerJ, 7, e7774. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Ma, Y.J. , He, H.P. , Zhao, H.M. , Xian, Y.D. , Guo, H. , Liu, B. et al. (2021) Microbiome diversity of cotton aphids (Aphis gossypii) is associated with host alternation. Scientific Reports, 11, 5260. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Matsumoto, Y. and Hattori, M. (2018) The green rice leafhopper, Nephotettix cincticeps (Hemiptera: Cicadellidae), salivary protein NcSP75 is a key effector for successful phloem ingestion. PLoS ONE, 13, e0202492. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Moriyama, M. , Nikoh, N. , Hosokawa, T. and Fukatsu, T. (2015) Riboflavin provisioning underlies Wolbachia’s fitness contribution to its insect host. mBio, 6, e01732–e01715. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Nakabachi, A. and Ishikawa, H. (1999) Provision of riboflavin to the host aphid, Acyrthosiphon pisum, by endosymbiotic bacteria, Buchnera . Journal of Insect Physiology, 45, 1–6. [DOI] [PubMed] [Google Scholar]
  47. Nalam, V. , Isaacs, T. , Moh, S. , Kansman, J. , Finke, D. , Albrecht, T. et al. (2021) Diurnal feeding as a potential mechanism of osmoregulation in aphids. Insect Science, 28, 521–532. [DOI] [PubMed] [Google Scholar]
  48. Nasreen, A. , Ghosh, R.K. , Ali, M. , Rahman, T. and Akhter, S. (2022) Estimation of thiamine, riboflavin and niacin content in jute leaves. GSC Advanced Research and Reviews, 13, 116–124. [Google Scholar]
  49. Nikolakakis, N.N. , Margaritopoulos, J.T. and Tsitsipis, J.A. (2003) Performance of Myzus persicae (Hemiptera: Aphididae) clones on different host‐plants and their host preference. Bulletin of Entomological Research, 93, 235–242. [DOI] [PubMed] [Google Scholar]
  50. Puterka, G.J. , Nicholson, S.J. and Cooper, W.R. (2017) Survival and feeding rates of four aphid species (Hemiptera: Aphididae) on various sucrose concentrations in diets. Journal of Economic Entomology, 110, 1518–1524. [DOI] [PubMed] [Google Scholar]
  51. Ray, D.M. and Savage, J.A. (2021) Seasonal changes in temperate woody plant phloem anatomy and physiology: implications for long‐distance transport. AoB Plants, 13, plab028. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Rihova, J.M. , Gupta, S. , Darby, A.C. , Novakova, E. and Hypsa, V. (2023) Arsenophonus symbiosis with louse flies: multiple origins, coevolutionary dynamics, and metabolic significance. mSystems, 8, 00706–00723. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Sadras, V. , Vazquez, C. , Garzo, E. , Moreno, A. , Medina, S. , Taylor, J. et al. (2021) The role of plant labile carbohydrates and nitrogen on wheat‐aphid relations. Scientific Reports, 11, 12529. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Sandhi, R.K. and Reddy, G.V.P. (2020) Biology, ecology, and management strategies for pea aphid (Hemiptera: Aphididae) in pulse crops. Journal of Integrated Pest Management, 11, 18. [Google Scholar]
  55. Santos‐Garcia, D. , Juravel, K. , Freilich, S. , Zchori‐Fein, E. , Latorre, A. , Moya, A. et al. (2018) To B or not to B: comparative genomics suggests Arsenophonus as a source of B vitamins in whiteflies. Frontiers in Microbiology, 9, 2254. [DOI] [PMC free article] [PubMed] [Google Scholar]
  56. Savkovic, U. , Dordevic, M. , Jovanovic, D.S. , Lazarevic, J. , Tucic, N. and Stojkovic, B. (2016) Experimentally induced host‐shift changes life‐history strategy in a seed beetle. Journal of Evolutionary Biology, 29, 837–847. [DOI] [PubMed] [Google Scholar]
  57. Shigenobu, S. , Watanabe, H. , Hattori, M. , Sakaki, Y. and Ishikawa, H. (2000) Genome sequence of the endocellular bacterial symbiont of aphids Buchnera sp APS. Nature, 407, 81–86. [DOI] [PubMed] [Google Scholar]
  58. Tian, P.P. , Chang, C.Y. , Miao, N.H. , Li, M.Y. and Liu, X.D. (2019) Infections with Arsenophonus facultative endosymbionts alter performance of aphids (Aphis gossypii) on an amino‐acid deficient diet. Applied and Environmental Microbiology, 85, e01407–e01419. [DOI] [PMC free article] [PubMed] [Google Scholar]
  59. Tian, P.P. , Zhang, Y.L. , Huang, J.L. , Li, W.Y. and Liu, X.D. (2023) Arsenophonus interacts with Buchnera to improve growth performance of aphids under amino acid stress. Microbiology Spectrum, 11, 01792–01723. [DOI] [PMC free article] [PubMed] [Google Scholar]
  60. Tsuchida, T. , Koga, R. and Fukatsu, T. (2004) Host plant specialization governed by facultative symbiont. Science, 303, 1989–1989. [DOI] [PubMed] [Google Scholar]
  61. Tsuchida, T. , Koga, R. , Matsumoto, S. and Fukatsu, T. (2011) Interspecific symbiont transfection confers a novel ecological trait to the recipient insect. Biology Letters, 7, 245–248. [DOI] [PMC free article] [PubMed] [Google Scholar]
  62. Via, S. (1991) Specialized host plant performance of pea aphid clones is not altered by experience. Ecology, 72, 1420–1427. [Google Scholar]
  63. Vilanova, C. , Baixeras, J. , Latorre, A. and Porcar, M. (2016) The generalist inside the specialist: gut bacterial communities of two insect species feeding on toxic plants are dominated by Enterococcus sp. Frontiers in Microbiology, 7, 1005. [DOI] [PMC free article] [PubMed] [Google Scholar]
  64. Vorburger, C. , Herzog, J. and Rouchet, R. (2017) Aphid specialization on different summer hosts is associated with strong genetic differentiation and unequal symbiont communities despite a common mating habitat. Journal of Evolutionary Biology, 30, 762–772. [DOI] [PubMed] [Google Scholar]
  65. Wagner, S.M. , Martinez, A.J. , Ruan, Y.M. , Kim, K.L. , Lenhart, P.A. , Dehnel, A.C. et al. (2015) Facultative endosymbionts mediate dietary breadth in a polyphagous herbivore. Function Ecology, 29, 1402–1410. [Google Scholar]
  66. Wang, G. , Zhou, J.J. , Li, Y. , Guo, Y. , Quandahor, P. and Liu, C. (2021a) Trehalose and glucose levels regulate feeding behavior of the phloem‐feeding insect, the pea aphid Acyrthosiphon pisum Harris. Scientific Reports, 11, 15864. [DOI] [PMC free article] [PubMed] [Google Scholar]
  67. Wang, L. , Zhang, S. , Luo, J.Y. , Wang, C.Y. , Lv, L.M. , Zhu, X.Z. et al. (2016) Identification of Aphis gossypii Glover (Hemiptera: Aphididae) biotypes from different host plants in North China. PLoS ONE, 11, e0146345. [DOI] [PMC free article] [PubMed] [Google Scholar]
  68. Wang, Y. , Eum, J.H. , Harrison, R.E. , Valzania, L. , Yang, X. , Johnson, J.A. et al. (2021b) Riboflavin instability is a key factor underlying the requirement of a gut microbiota for mosquito development. Proceedings of the National Academy of Sciences USA, 118, e2101080118. [DOI] [PMC free article] [PubMed] [Google Scholar]
  69. Wang, Y.B. , Ren, F.R. , Yao, Y.L. , Sun, X. , Walling, L.L. , Li, N.N. et al. (2020) Intracellular symbionts drive sex ratio in the whitefly by facilitating fertilization and provisioning of B vitamins. The ISME Journal, 14, 2923–2935. [DOI] [PMC free article] [PubMed] [Google Scholar]
  70. Wang, Z.W. , Zhao, J. , Li, G.Y. , Hu, D. , Wang, Z.G. , Ye, C. et al. (2024) The endosymbionts Serratia symbiotica improves aphid fitness by disrupting the predation strategy of ladybeetle larvae. Insect Science, 31, 1555–1568. [DOI] [PubMed] [Google Scholar]
  71. Wetzel, W.C. , Kharouba, H.M. , Robinson, M. , Holyoak, M. and Karban, R. (2016) Variability in plant nutrients reduces insect herbivore performance. Nature, 539, 425–427. [DOI] [PubMed] [Google Scholar]
  72. Wilkinson, T. , Ashford, D. , Pritchard, J. and Douglas, A. (1997) Honeydew sugars and osmoregulation in the pea aphid Acyrthosiphon pisum . Journal of Experimental Biology, 200, 2137–2143. [DOI] [PubMed] [Google Scholar]
  73. Winter, H. , Lohaus, G. and Heldt, H.W. (1992) Phloem transport of amino acids in relation to their cytosolic levels in barley leaves. Plant Physiology, 99, 996–1004. [DOI] [PMC free article] [PubMed] [Google Scholar]
  74. Wu, W. , Liang, X.L. , Zhao, H.Y. , Xu, T.T. and Liu, X.D. (2013) Special plant species determines diet breadth of phytophagous insects: a study on host plant expansion of the host‐specialized Aphis gossypii Glover. PLoS ONE, 8, e60832. [DOI] [PMC free article] [PubMed] [Google Scholar]
  75. Wulff, J.A. and White, J.A. (2015) The endosymbiont Arsenophonus provides a general benefit to soybean aphid (Hemiptera: Aphididae) regardless of host plant resistance (Rag). Environmental Entomology, 44, 574–581. [DOI] [PubMed] [Google Scholar]
  76. Xu, S. , Jiang, L. , Qiao, G. and Chen, J. (2020) The bacterial flora associated with the polyphagous aphid Aphis gossypii Glover (Hemiptera: Aphididae) is strongly affected by host plants. Microbial Ecology, 79, 971–984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  77. Xu, T.T. , Ma, T.T. and Liu, X.D. (2014) How does the host‐specialized aphid deal with food deficiency? Insect Science, 21, 334–341. [DOI] [PubMed] [Google Scholar]
  78. Xu, W. , Liu, W. , Li, J. , Zhu, X. , Wang, L. , Li, D. et al. (2023) Buchnera breaks the specialization of the cotton‐specialized aphid (Aphis gossypii) by providing nutrition through zucchini. Frontiers in Nutrition, 10, 1128272. [DOI] [PMC free article] [PubMed] [Google Scholar]
  79. Yang, C. , Pan, H. , Liu, Y. and Zhou, X. (2014) Selection of reference genes for expression analysis using quantitative real‐time PCR in the pea aphid, Acyrthosiphon pisum (Harris) (Hemiptera, Aphidiae). PLoS ONE, 9, e110454. [DOI] [PMC free article] [PubMed] [Google Scholar]
  80. Yorimoto, S. , Hattori, M. , Kondo, M. and Shigenobu, S. (2022) Complex host/symbiont integration of a multi‐partner symbiotic system in the eusocial aphid Ceratovacuna japonica . iScience, 25, 105478. [DOI] [PMC free article] [PubMed] [Google Scholar]
  81. Zhang, Y.C. , Cao, W.J. , Zhong, L.R. , Godfray, H.C.J. and Liu, X.D. (2016) Host plant determines the population size of an obligate symbiont (Buchnera aphidicola) in aphids. Applied and Environmental Microbiology, 82, 2336–2346. [DOI] [PMC free article] [PubMed] [Google Scholar]

Associated Data

This section collects any data citations, data availability statements, or supplementary materials included in this article.

Supplementary Materials

Fig. S1 The sequences of 6 riboflavin genes of Buchnera.

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Fig. S2 Net reproductive rate and average generational lifespan of CA3 populations infected with Arsenophonus (Ars+) or not (Ars‐) on the diet with riboflavin or not.

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Table S1 Nutritional composition of the artificial diet with 15% sucrose.

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Table S2 Nutritional composition of the artificial diet with 25 mg/L riboflavin.

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Table S3 Primers used in this study.

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