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ABSTRACT Cotton–melon aphid (Aphis gossypii) is a highly destructive pest that causes serious yield losses in cucumber production. IL52, a Cucumis hystrix introgression line of cucumber, exhibits resistance to a number of diseases and also shows strong resistance to aphids. To characterize the type of aphid resistance in IL52, we designed a separate leaf‐disc/no‐choice test and free‐choice test to examine antibiosis and antixenosis, respectively. Our results indicated that IL52 displayed antixenosis resistance to aphids. In other words, when planted with other lines such as CCMC, IL52 was not favourable to aphids. To dissect the genetic basis of antixenosis resistance in IL52, we evaluated a mapping population of 155 lines from CCMC × IL52–derived F7:8 RIL using free‐choice test for aphid resistance. A major‐effect QTL on Chr2, designated as qRag2.1 (Resistant to A. gossypii), was identified as being significantly associated with aphid resistance. The QTL qRag2.1 was mapped to an interval flanked by markers UW085197 and SSR11909, spanning a physical location of 15.14–19.07 Mb, which explained 12.74%–14.03% of the phenotypic variance. Analysis of polymorphisms between the parental lines using resequencing data revealed one gene, CsaV3_2G018180, with three nonsynonymous amino acid substitutions and a 6‐bp InDel in its coding sequence (CDS). CsaV3_2G018180 encodes a pectinesterase/pectinesterase inhibitor enzyme, designated as CsPEI, which has been reported to regulate pectin methylesterase activity that is implicated in various biological processes. The relative expression level of CsPEI was induced in IL52 post aphid infestation, suggesting it could be a potential candidate gene for aphid resistance. [ABSTRACT FROM AUTHOR]

Ecdysone receptor (EcR) plays important roles in molting, metamorphosis, and reproduction of insects. EcR ortholog and its function in melon aphid Aphis gossypii have not been documented yet. RNA interference (RNAi) is a reverse-genetic approach used to study functional genomics, but recently, it has also shown promise as a novel tool that could be used to control insect pests. In this study, reverse transcription-PCR (RT-PCR), quantitative real-time PCR (qPCR), RNAi, and bioassays were employed to isolate EcR gene and investigate its knock-down effects on aphids. The lethal and sublethal effects were also studied after silencing the EcR gene. The results showed that EcR gene is highly expressed (P < 0.006) in 2nd nymphal stage of A. gossypii while expression was statistically lower in all other developmental stages. Knock-down of EcR gene caused up to 43 and 68% mortality in 2nd instar nymphs after feeding dsEcR for 48 and 72 h, respectively, compared to the control. Moreover, RNAi-induced silencing of EcR gene caused sublethal effects on surviving aphids. Significantly lower longevity (about 42%) and fecundity (about 44.22%) were observed in insects fed with dsEcR solution at their nymphal stage. Analysis of gene expression showed that the mortality rates and the lowered longevity and fecundity were attributable to the down-regulation of EcR gene by RNA interference technology. Taken together, our results demonstrate that EcR is an essential gene for aphid growth and development and could be a potential target gene for the development of an RNAi-based bio-aphicide. [ABSTRACT FROM AUTHOR]

The complete mitochondrial genome of Aphis gossypii Glover cucumber biotype was sequenced using traditional PCR amplification coupled with Sanger sequencing. The genome is 15,870 bp long, with 83.7% AT content (MW048625). The genome encodes 37 typical mitochondrial genes, including 13 protein-coding genes, 2 ribosomal RNA genes, 22 transfer RNAs, a repeat region of 784 bp, and a control region of 627 bp. The base composition of the genome is A (45.4%), T (38.3%), C (10.5%), and G (5.8%). An analysis of two biotypes A. gossypii mitogenomes identified 77 single nucleotide polymorphisms and 1 insertion and deletion.

Aphis gossypii is a globally distributed species and therefore has a highly variable life cycle. Populations of A. gossypii in northern China exhibit greater genotypic diversity and a broader host range, yet the details of life cycles of different biotypes is still unclear. In this study, the Cytb and 16S gene regions of A. gossypii collected from 5 common summer hosts and 4 primary hosts were analyzed. A total of 57 haplotypes were obtained from 1 046 individual A. gossypii sequences. The sequence included 44 variable sites, 27 of which were parsimony informative sites and 17 of which were singleton variable sites. The most frequent 3 haplotypes were found in 896 individuals, representing a total of 85.7% of all individuals and 36 haplotypes were found in 1 individual. A neighbor-joining tree was constructed using 21 haplotypes that were found in more than 2 individuals. Considering the individual host plant, 5 biotypes were identified. Type 1 corresponded exactly to the cucurbit host-race and the other 4 biotypes were found as cotton host-races. Type 3 was the most abundant biotype in cotton fields in northern China.

Background A polyphagous insect herbivore has a wide range of host plants. However, it has been found that many polyphagous herbivores commonly exhibit a strong preference for a subset of species in their broad host range, and various host biotypes exist in herbivore populations. Nutrition and secondary metabolites in plants affect herbivore preference and performance, but it is still not clear which factors determine the host range and host preference of polyphagous herbivores. Method Cotton-melon aphids, Aphis gossypii Glover, collected from cotton and cucumber crops, were used in this study. The genetic backgrounds of these aphids were detected using microsatellite PCR and six genotypes were evaluated. Performance of these six aphid genotypes on excised leaves and plants of cotton and cucumber seedlings were examined through a reciprocal transplant experiment. In order to detect whether the feeding experience on artificial diet would alter aphid host range, the six genotypes of aphids fed on artificial diet for seven days were transferred onto cotton and cucumber leaves, and then their population growth on these two host plants was surveyed. Results Aphids from cotton and cucumber plants could not colonize the excised leaves and intact plants of cucumber and cotton seedlings, respectively. All six genotypes of aphids collected from cotton and cucumber plants could survive and produce offspring on artificial diet, which lacked plant secondary metabolites. The feeding experience on the artificial diet did not alter the ability of all six genotypes to use their native host plants. However, after feeding on this artificial diet for seven days, two aphid genotypes from cotton and one from cucumber acquired the ability to use both of the excised leaves from cucumber and cotton plants. The two aphid genotypes from cotton conditioned by the feeding experience on artificial diet and then reared on excised cucumber leaves for >12 generations still maintained the ability to use intact cotton plants but did not establish a population on cucumber plants. However, one cucumber genotype conditioned by artificial diet and then reared on excised cotton leaves could use both the intact cotton and cucumber plants, showing that the expansion of host range was mediated by feeding experience. Conclusion Feeding experience on artificial diet induced the expansion of host range of the cucurbit-specialized A. gossypii, and this expansion was genotype-specific. We speculated that feeding on a constant set of host plants in the life cycle of aphids may contribute to the formation of host specialization.

Chitin is a vital part of the insect exoskeleton and peritrophic membrane, synthesized by chitin synthase (CHS) enzymes. Chitin synthase 1 (CHS1) is a crucial enzyme in the final step of chitin biosynthetic pathway and consequently plays essential role towards insect growth and molting. RNA interference (RNAi) is an agent that could be used as an extremely target-specific and ecologically innocuous tactic to control different insect pests associated with economically important crops. The sole purpose of the current study is to use CHS1 as the key target gene against the cotton-melon aphid, Aphis gossypii, via oral feeding on artificial diets mixed with dsRNA-CHS1. Results revealed that the expression level of CHS1 gene significantly decreased after the oral delivery of dsRNA-CHS1. The knockdown of CHS1 gene caused up to 43%, 47%, and 59% mortality in third-instar nymph after feeding of dsCHS1 for 24, 48, and 72 h, respectively, as compared to the control. Consistent with this, significantly lower longevity (approximately 38%) and fecundity (approximately 48%) were also found in adult stage of cotton-melon aphids that were fed with dsCHS1 for 72 h at nymphal stage. The qRT-PCR analysis of gene expression demonstrated that the increased mortality rates and lowered longevity and fecundity of A. gossypii were attributed to the downregulation of CHS1 gene via oral-delivery-mediated RNAi. The results of current study confirm that CHS1 could be an appropriate candidate target gene for the RNAi-based control of cotton-melon aphids.

Host specialization is an ubiquitous character in aphid populations. Many polyphagous aphid populations usually consist of several subpopulations that have strong fidelity to a specific host or a subset of host range. Host specialization is an evolutional result of food habit of insects. However, genetic basis and molecular mechanism of host specialization are still unclear. In this study, we presented a comparative analysis on global gene expression profiles of three lineages of Aphis gossypii Glover: cotton-specialized (CO), cucurbit-specialized (CU), and CU reared on cowpea (CU-cowpea), using RNA-Seq method. More than 157 million clean reads and 38,398 different unigenes were generated from transcriptomes of these three aphid lineages. The 1,106 down- and 2,835 up-regulated genes were found between CO and CU, and 812 down- and 14,492 up-regulated genes between CU-cowpea and CU. Differentially expressed genes between CO and CU were enriched in sugar metabolism, immune system process, pathogen infection or symbiosis, and salivary secretion. Genes associated with cytochrome P450, major facilitator superfamily, and salivary effector were differentially expressed between CO and CU, which might be involved in determining host specialization. UDP-glycosyltransferases genes were sensitive to host shift. Carboxylesterases and digestion-related protease genes were related to both the host specialization and host shift of aphids. Expression levels of 22 out of 24 genes of CO and CU measured by RT-qPCR method were as similar as the results from RNA-seq method. This study provides a road map for future study on molecular mechanism of host specialization in aphids. [ABSTRACT FROM AUTHOR]

The melon aphid, Aphis gossypii Glover (Hemiptera: Aphididae), is a cosmopolitan polyphagous aphid species that inflicts serious damage to a wide spectrum of crops and has an extraordinary ability to transmit viruses in either a nonpersistent or persistent mode. It tends to specialize on a few host plants species, resulting in parapatric divergence among populations in a given geographical area over a period of time. This seems to be the major reason for its remarkable diversity resulting in the evolution of biotypes and cryptic species favoring host adaptation and reproductive isolation across various localities. This notorious pest is supposed to have a number of putative biotypes associated with hosts or geographical locations. It is suspected that A. gossypii occurring on cotton (Malvaceae) and melons (Cucurbitaceae) are different biotypes. The relationship between host or geographic preference and genetic variation of aphids is unclear and thus requires further investigation. In light of this, the present study was conducted to determine if host- or geographical location-associated genetic differences are indeed present in 116 populations of A. gossypii collected from various agroclimatic zones of India on various host plants with special emphasis on Malvaceae and Cucurbitaceae families using the well-known mitochondrial cytochrome oxidase I (mtCOI) marker. Alignment of mtCOI sequences of A. gossypii along with available sequences in the National Center for Biotechnology Information (NCBI), GenBank, and Barcode of Life Data (BOLD) system revealed maximum sequence identity of 99% with very few variable sites indicating an extremely low level of intraspecific variation. The phylogram strongly suggested that there is no major host- or geographical location-associated genetic differences in A. gossypii, and it is a single cosmopolitan species devoid of genetic variations. This study aids to resolve evolutionary relationships among closely related species groups and detect newly evolving biotypes. [ABSTRACT FROM AUTHOR]

Aphis gossypii Glover shows obvious host specialization, with cucurbit- and cotton-specialized biotypes or host races in many regions. Because its annual natal host crops senesce earlier the cucurbit-specialized biotype may suffer food deficiency. The method this biotype uses to overcome this challenge is still poorly understood. In order to understand the potential of the cucurbit-specialized biotype aphids in host shift and usage, the performance of this biotype on cotton ( Gossypium hirsutum), a common but poor quality host plant, was explored in this study. The cucurbit-specialized aphids could establish populations on cotton only when these plants had at least nine leaves, and subsequent populations developed rather slowly. The presence of whitefly populations on cotton improved the success rate of cucurbit-specialized aphids. The cucurbit-specialized aphids were mainly distributed on the older leaves of cotton, with only a few settling on the upper leaves. The cucurbit-specialized aphids reared on cotton for 40, 54 and 61 days still maintained strong preference for their natal host plant, cucumber ( Cucumis sativus), rather than cotton, and their net reproductive rates and intrinsic rates of natural increase were dramatically lower when they were transferred onto new six-leaf cotton plants or detached leaves. Therefore, we concluded that the cucurbit-specialized aphids have the potential to utilize mature or whitefly-stressed cotton plants, but that this feeding experience on cotton did not alter their specialization for cucurbits. Some cotton plants could act as a temporary host for the cucurbit-specialized aphids to overcome food deficiency arising from senescing cucurbits. [ABSTRACT FROM AUTHOR]

Although distinct host specialization is observed for the cotton-melon aphid ( Aphis gossypii Glover) on cotton and cucurbit plants, it is still ambiguous whether the specialization is altered by experience on a novel host plant. Here the performance of cotton and cucurbit-specialized aphids, A. gossypii on novel host plants was studied by a host-selection test and by the life-table method. The two host-specialized aphids cannot survive and establish populations after reciprocal host transfers. They have ability to recognize the host plants on which they were reared, and escape behavior from novel hosts was observed. Interestingly, the cotton and cucurbit-specialized aphids survive and reproduce normally on hibiscus ( Hibiscus syriacus), a main overwintering host plant, and host-fidelity of A. gossypii to cucurbit plants is altered by feeding and living experience on hibiscus, which confers the same capacity to use cotton and cucumber on to the cucurbit-specialized population, but host-fidelity to cotton is not altered and the fitness of the cotton specialized population to cucumber is still poorer. A. gossypii from hibiscus has a significant preference for cotton to cucumber in the host-selection process, and none stays on cucumber more than 20 h after transfer. The results presented imply that cucurbit-specialized aphids might not return to an overwintering host plant (hibiscus) in wild fields, so host conservatism to cucurbit plants is maintained. The potential of cucurbit-specialized aphids of A. gossypii to use cotton plants, intermediated by experience on hibiscus, suggests that the specialized host-plant performance of phytophagous insects is not wholly conservative. [ABSTRACT FROM AUTHOR]

The melon aphid, Aphis gossypii is a globally distributed crop pest with a wide host range. The intensive use of insecticides against this insect over several years has led to develop resistance against many insecticides including acetamiprid. Understanding the relationship between acetamiprid resistance and fitness of A. gossypii is essential to limit the spread of the resistant population in the field. In this study, age-stage, two-sex life table approach was used to investigate these relationships in the lab. Results showed that resistant strain (Ace-R) had a reduced fitness (relative fitness = 0.909) along with significantly decreased adult longevity, fecundity, net reproductive (R 0), mean generation time (T) and gross reproductive rate (GRR). Compared to the susceptible strain (Ace-S), the pre-adult period and total pre-oviposition period (TPOP) were also significantly shorter in Ace-R strain. Moreover, the expression profiles of EcR , JHBP , JHAMT , JHEH, USP and Vg genes supposed to be involved in insect reproduction and development were analyzed using Quantitative Real Time PCR. The EcR , JHBP , JHAMT and USP genes were up-regulated, Vg gene was down-regulated while the mRNA level of JHEH gene was statistically same in the Ace-R strain compared to the Ace-S strain. Collectively, this study provides the occurrence and magnitude of fitness costs of A. gossypii against acetamiprid resistance and could be helpful to manage the resistance evolution in field populations. Unlabelled Image • Age-stage, two-sex life table approach was used to investigate the fitness of acetamiprid-resistant (Ac-R) and susceptible (Ace-S) strains of A. gossypii. • Acetamiprid resistance development reduced the survival rate and reproductive capacity of resistant A. gossypii. • The Ace-R strain suffered reduction in demographic parameters and relative fitness. • Up-regulation of EcR , JHBP , JHAMT, and USP , and down-regulation of Vg in Ace-R may have important role in observed trade-off. [ABSTRACT FROM AUTHOR]

Chitin is a vital part of the insect exoskeleton and peritrophic membrane, synthesized by chitin synthase (CHS) enzymes. Chitin synthase 1 (CHS1) is a crucial enzyme in the final step of chitin biosynthetic pathway and consequently plays essential role towards insect growth and molting. RNA interference (RNAi) is an agent that could be used as an extremely target-specific and ecologically innocuous tactic to control different insect pests associated with economically important crops. The sole purpose of the current study is to use CHS1 as the key target gene against the cotton-melon aphid, Aphis gossypii, via oral feeding on artificial diets mixed with dsRNA-CHS1. Results revealed that the expression level of CHS1 gene significantly decreased after the oral delivery of dsRNA-CHS1. The knockdown of CHS1 gene caused up to 43%, 47%, and 59% mortality in third-instar nymph after feeding of dsCHS1 for 24, 48, and 72 h, respectively, as compared to the control. Consistent with this, significantly lower longevity (approximately 38%) and fecundity (approximately 48%) were also found in adult stage of cotton-melon aphids that were fed with dsCHS1 for 72 h at nymphal stage. The qRT-PCR analysis of gene expression demonstrated that the increased mortality rates and lowered longevity and fecundity of A. gossypii were attributed to the downregulation of CHS1 gene via oral-delivery-mediated RNAi. The results of current study confirm that CHS1 could be an appropriate candidate target gene for the RNAi-based control of cotton-melon aphids. [ABSTRACT FROM AUTHOR]

The complete mitochondrial genome of Aphis gossypii Glover cucumber biotype was sequenced using traditional PCR amplification coupled with Sanger sequencing. The genome is 15,870 bp long, with 83.7% AT content (MW048625). The genome encodes 37 typical mitochondrial genes, including 13 protein-coding genes, 2 ribosomal RNA genes, 22 transfer RNAs, a repeat region of 784 bp, and a control region of 627 bp. The base composition of the genome is A (45.4%), T (38.3%), C (10.5%), and G (5.8%). An analysis of two biotypes A. gossypii mitogenomes identified 77 single nucleotide polymorphisms and 1 insertion and deletion., Competing Interests: No potential conflict of interest was reported by the author(s)., (© 2021 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.)

Chitin is a vital part of the insect exoskeleton and peritrophic membrane, synthesized by chitin synthase (CHS) enzymes. Chitin synthase 1 (CHS1) is a crucial enzyme in the final step of chitin biosynthetic pathway and consequently plays essential role towards insect growth and molting. RNA interference (RNAi) is an agent that could be used as an extremely target-specific and ecologically innocuous tactic to control different insect pests associated with economically important crops. The sole purpose of the current study is to use CHS1 as the key target gene against the cotton-melon aphid, Aphis gossypii , via oral feeding on artificial diets mixed with dsRNA- CHS1 . Results revealed that the expression level of CHS1 gene significantly decreased after the oral delivery of dsRNA- CHS1 . The knockdown of CHS1 gene caused up to 43%, 47%, and 59% mortality in third-instar nymph after feeding of ds CHS1 for 24, 48, and 72 h, respectively, as compared to the control. Consistent with this, significantly lower longevity (approximately 38%) and fecundity (approximately 48%) were also found in adult stage of cotton-melon aphids that were fed with ds CHS1 for 72 h at nymphal stage. The qRT-PCR analysis of gene expression demonstrated that the increased mortality rates and lowered longevity and fecundity of A. gossypii were attributed to the downregulation of CHS1 gene via oral-delivery-mediated RNAi. The results of current study confirm that CHS1 could be an appropriate candidate target gene for the RNAi-based control of cotton-melon aphids.

Background: A polyphagous insect herbivore has a wide range of host plants. However, it has been found that many polyphagous herbivores commonly exhibit a strong preference for a subset of species in their broad host range, and various host biotypes exist in herbivore populations. Nutrition and secondary metabolites in plants affect herbivore preference and performance, but it is still not clear which factors determine the host range and host preference of polyphagous herbivores., Method: Cotton-melon aphids, Aphis gossypii Glover, collected from cotton and cucumber crops, were used in this study. The genetic backgrounds of these aphids were detected using microsatellite PCR and six genotypes were evaluated. Performance of these six aphid genotypes on excised leaves and plants of cotton and cucumber seedlings were examined through a reciprocal transplant experiment. In order to detect whether the feeding experience on artificial diet would alter aphid host range, the six genotypes of aphids fed on artificial diet for seven days were transferred onto cotton and cucumber leaves, and then their population growth on these two host plants was surveyed., Results: Aphids from cotton and cucumber plants could not colonize the excised leaves and intact plants of cucumber and cotton seedlings, respectively. All six genotypes of aphids collected from cotton and cucumber plants could survive and produce offspring on artificial diet, which lacked plant secondary metabolites. The feeding experience on the artificial diet did not alter the ability of all six genotypes to use their native host plants. However, after feeding on this artificial diet for seven days, two aphid genotypes from cotton and one from cucumber acquired the ability to use both of the excised leaves from cucumber and cotton plants. The two aphid genotypes from cotton conditioned by the feeding experience on artificial diet and then reared on excised cucumber leaves for >12 generations still maintained the ability to use intact cotton plants but did not establish a population on cucumber plants. However, one cucumber genotype conditioned by artificial diet and then reared on excised cotton leaves could use both the intact cotton and cucumber plants, showing that the expansion of host range was mediated by feeding experience., Conclusion: Feeding experience on artificial diet induced the expansion of host range of the cucurbit-specialized A. gossypii , and this expansion was genotype-specific. We speculated that feeding on a constant set of host plants in the life cycle of aphids may contribute to the formation of host specialization., Competing Interests: The authors declare there are no competing interests., (©2019 Ma et al.)

The cotton-melon aphid, Aphis gossypii Glover, is a worldwide-spreading species, and pesticide-resistant populations are increasing rapidly. In this study, investigations were performed based on Illumina HiSeq sequencing of the 16S rDNA V4 region for the bacterial communities embodied as intracellular symbionts under natural and in pesticide-treated populations of A. gossypii. The results revealed that more than 82% of bacterial communities belonged to the phylum Proteobacteria in which the maximum proportion (53.24%) was of the genus Arsenophonus; Hamiltonella composed 22.31; and 1.37% was of the genus Acinetobacter. The relative abundance of Hamiltonella was obvious, vertically transmitted, divided into two groups, and its infection influenced the bacterial communities in A. gossypii. Symbiont density and composition were changed in samples tested on different days. Azadirachtin and phoxim influenced on the composition of bacterial communities. Different biomarkers were used for pesticide-treated samples with LEfSe results. These findings will increase awareness regarding bacterial communities in naturally occurring populations of A. gossypii and pave the way to study the relationship between symbionts and pesticide resistance., (© 2018 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.)

Host specialization is an ubiquitous character in aphid populations. Many polyphagous aphid populations usually consist of several subpopulations that have strong fidelity to a specific host or a subset of host range. Host specialization is an evolutional result of food habit of insects. However, genetic basis and molecular mechanism of host specialization are still unclear. In this study, we presented a comparative analysis on global gene expression profiles of three lineages of Aphis gossypii Glover: cotton-specialized (CO), cucurbit-specialized (CU), and CU reared on cowpea (CU-cowpea), using RNA-Seq method. More than 157 million clean reads and 38,398 different unigenes were generated from transcriptomes of these three aphid lineages. The 1,106 down- and 2,835 up-regulated genes were found between CO and CU, and 812 down- and 14,492 up-regulated genes between CU-cowpea and CU. Differentially expressed genes between CO and CU were enriched in sugar metabolism, immune system process, pathogen infection or symbiosis, and salivary secretion. Genes associated with cytochrome P450, major facilitator superfamily, and salivary effector were differentially expressed between CO and CU, which might be involved in determining host specialization. UDP-glycosyltransferases genes were sensitive to host shift. Carboxylesterases and digestion-related protease genes were related to both the host specialization and host shift of aphids. Expression levels of 22 out of 24 genes of CO and CU measured by RT-qPCR method were as similar as the results from RNA-seq method. This study provides a road map for future study on molecular mechanism of host specialization in aphids., (© The Authors 2017. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)