Aphids are phloem-feeding insects that reduce crop productivity due to feeding and transmission of plant viruses.When aphids disperse across the landscape to colonize new host plants,they will often probe on a wide va...Aphids are phloem-feeding insects that reduce crop productivity due to feeding and transmission of plant viruses.When aphids disperse across the landscape to colonize new host plants,they will often probe on a wide variety of nonhost plants before settling on a host suitable for feeding and reproduction.There is limited understanding of the diversity of plants that aphids probe on within a landscape,and characterizing this diversity can help us better understand host use patterns of aphids.Here,we used gut content analysis(GCA)to identify plant genera that were probed by aphid vectors of potato virus Y(PVY).Aphids were trapped weekly near potato fields during the growing seasons of 2020 and 2021 in San Luis Valley in Colorado.High-throughput sequencing of plant barcoding genes,trnF and ITS2,from 200 individual alate(i.e.,winged)aphids representing nine vector species of PVY was performed using the PacBio sequencing platform,and sequences were identi-fied to genus using NCBI BLASTn.We found that 34.7%of aphids probed upon presumed PVY host plants and that two of the most frequently detected plant genera,Solanum and Brassica,represent important crops and weeds within the study region.We found that 75%of aphids frequently probed upon PVY nonhosts including many species that are outside of their reported host ranges.Additionally,19%of aphids probed upon more than one plant species.This study provides the first evidence from high-throughput molecular GCA of aphids and reveals host use patterns that are relevant for PVY epidemiology.展开更多
Gut content analysis is a useful tool when studying arthropod predator-prey interactions. We used polymerase chain reaction （PCR） technique to examine how detection of prey DNA in the gut content of predators was in...Gut content analysis is a useful tool when studying arthropod predator-prey interactions. We used polymerase chain reaction （PCR） technique to examine how detection of prey DNA in the gut content of predators was influenced by digestion time and temperature. Such knowledge is critical before applying PCR-based gut content analysis to field collected predators. Larvae of the two-spotted ladybeetle （Adalia bipunctata L.） were fed with the bird cherry-oat aphid （Rhopalosiphum padi L.） at either 21℃ or 14℃. After consuming one aphid, the predators were allowed to digest the prey for a range of time periods up to 24 hours. The influence of temperature on A. bipunctata feeding behavior was also recorded. From the fed larvae, total DNA was extracted and PCR reactions with R. padi specific primers were run. The number ofA. bipunctata that tested positive for R. padi DNA was negatively related to the length of digestion time. Temperature influenced larval feeding behavior but did not have a significant effect on R. padi DNA detection. After pooling the data from both temperature treatments we estimated the time point when R. padi DNA could be amplified from 50% of the fed A. bipunctata by PCR to be 4.87 hours. With such a rapid decrease in prey DNA detection success, positive PCR reactions will most likely be the result of predation events occurring shortly before capture. If a defined digestion temperature range has proven not to influence prey detection, PCR data obtained from predators collected within that particular range can be interpreted in the same way.展开更多
Floodwater mosquitoes （Diptera： Culicidae） are associated with periodically flooded wet meadows, marshes, and swamps in floodplains of major rivers worldwide, and their larvae are abundant in the shallow parts of f...Floodwater mosquitoes （Diptera： Culicidae） are associated with periodically flooded wet meadows, marshes, and swamps in floodplains of major rivers worldwide, and their larvae are abundant in the shallow parts of flooded areas. The nuisance caused by the blood-seeking adult female mosquitoes motivates mosquito control. Larviciding with Bacillus thuringiensis israelensis is considered the most environmentally safe method. However, some concern has been raised whether aquatic predatory insects could be indi- rectly affected by this reduction in a potential vital prey. Top predators in the temporary wetlands in the River Dal/ilven floodplains are diving beetles （Coleoptera： Dytiscidae）, and Aedes sticticus andAe, vexans are the target species for mosquito control. For detailed stud- ies on this aquatic predator-prey system, we developed a polymerase chain reaction （PCR） assay for detection of mosquito DNA in the guts of medinm-sized diving beetles. Primers were designed for amplifying short mitochondrial DNA fragments of the cytochrome C oxidase subunit I （COI） gene in Ae. sticticus and Ae. vexans, respectively. Primer speci- ficity was confirmed and half-life detectability ofAe. sticticus DNA in diving beetle guts was derived from a feeding and digestion experiment. The Ae. sticticus DNA within diving beetle guts was detected up to 12 h postfeeding, and half-life detectability was estimated to 5.6 h. In addition, field caught diving beetles were screened for Ae. sticticus and Ae. vexans DNA and in 14% of the diving beetles one or both mosquito species were detected, showing that these mosquito species are utilized as food by the diving beetles.展开更多
文摘Aphids are phloem-feeding insects that reduce crop productivity due to feeding and transmission of plant viruses.When aphids disperse across the landscape to colonize new host plants,they will often probe on a wide variety of nonhost plants before settling on a host suitable for feeding and reproduction.There is limited understanding of the diversity of plants that aphids probe on within a landscape,and characterizing this diversity can help us better understand host use patterns of aphids.Here,we used gut content analysis(GCA)to identify plant genera that were probed by aphid vectors of potato virus Y(PVY).Aphids were trapped weekly near potato fields during the growing seasons of 2020 and 2021 in San Luis Valley in Colorado.High-throughput sequencing of plant barcoding genes,trnF and ITS2,from 200 individual alate(i.e.,winged)aphids representing nine vector species of PVY was performed using the PacBio sequencing platform,and sequences were identi-fied to genus using NCBI BLASTn.We found that 34.7%of aphids probed upon presumed PVY host plants and that two of the most frequently detected plant genera,Solanum and Brassica,represent important crops and weeds within the study region.We found that 75%of aphids frequently probed upon PVY nonhosts including many species that are outside of their reported host ranges.Additionally,19%of aphids probed upon more than one plant species.This study provides the first evidence from high-throughput molecular GCA of aphids and reveals host use patterns that are relevant for PVY epidemiology.
文摘Gut content analysis is a useful tool when studying arthropod predator-prey interactions. We used polymerase chain reaction （PCR） technique to examine how detection of prey DNA in the gut content of predators was influenced by digestion time and temperature. Such knowledge is critical before applying PCR-based gut content analysis to field collected predators. Larvae of the two-spotted ladybeetle （Adalia bipunctata L.） were fed with the bird cherry-oat aphid （Rhopalosiphum padi L.） at either 21℃ or 14℃. After consuming one aphid, the predators were allowed to digest the prey for a range of time periods up to 24 hours. The influence of temperature on A. bipunctata feeding behavior was also recorded. From the fed larvae, total DNA was extracted and PCR reactions with R. padi specific primers were run. The number ofA. bipunctata that tested positive for R. padi DNA was negatively related to the length of digestion time. Temperature influenced larval feeding behavior but did not have a significant effect on R. padi DNA detection. After pooling the data from both temperature treatments we estimated the time point when R. padi DNA could be amplified from 50% of the fed A. bipunctata by PCR to be 4.87 hours. With such a rapid decrease in prey DNA detection success, positive PCR reactions will most likely be the result of predation events occurring shortly before capture. If a defined digestion temperature range has proven not to influence prey detection, PCR data obtained from predators collected within that particular range can be interpreted in the same way.
文摘Floodwater mosquitoes （Diptera： Culicidae） are associated with periodically flooded wet meadows, marshes, and swamps in floodplains of major rivers worldwide, and their larvae are abundant in the shallow parts of flooded areas. The nuisance caused by the blood-seeking adult female mosquitoes motivates mosquito control. Larviciding with Bacillus thuringiensis israelensis is considered the most environmentally safe method. However, some concern has been raised whether aquatic predatory insects could be indi- rectly affected by this reduction in a potential vital prey. Top predators in the temporary wetlands in the River Dal/ilven floodplains are diving beetles （Coleoptera： Dytiscidae）, and Aedes sticticus andAe, vexans are the target species for mosquito control. For detailed stud- ies on this aquatic predator-prey system, we developed a polymerase chain reaction （PCR） assay for detection of mosquito DNA in the guts of medinm-sized diving beetles. Primers were designed for amplifying short mitochondrial DNA fragments of the cytochrome C oxidase subunit I （COI） gene in Ae. sticticus and Ae. vexans, respectively. Primer speci- ficity was confirmed and half-life detectability ofAe. sticticus DNA in diving beetle guts was derived from a feeding and digestion experiment. The Ae. sticticus DNA within diving beetle guts was detected up to 12 h postfeeding, and half-life detectability was estimated to 5.6 h. In addition, field caught diving beetles were screened for Ae. sticticus and Ae. vexans DNA and in 14% of the diving beetles one or both mosquito species were detected, showing that these mosquito species are utilized as food by the diving beetles.
