Quantitative reverse transcription polymerase chain reaction(qRT-PCR) is the most commonly-used tool for measurement of gene expression, but its accuracy and reliability depend on appropriate data normalization with t...Quantitative reverse transcription polymerase chain reaction(qRT-PCR) is the most commonly-used tool for measurement of gene expression, but its accuracy and reliability depend on appropriate data normalization with the use of one or more stable reference genes. Adelphocoris suturalis is one of the most destructive pests of cotton, but until recently knowledge of its underlying molecular physiology had been hindered by a lack of molecular resources. To facilitate research on this pest, we evaluated 12 common housekeeping genes studied in insects(GAPDH, ACT, βACT, TBP, SDH, βTUB, EF1γ, EF1α, EF1δ, RPL32, RPS15, and RPL27) for their expression stability in A. suturalis when subjected to various experimental treatments, including three biotic(developmental stage and sex, tissue type, and metathoracic scent gland for varying developmental stages and sexes) and one abiotic(RNA interference injection) conditions. Four dedicated algorithms(ΔCt method, geNorm, BestKeeper and NormFinder) were used to analyze gene expression stability. In addition, RefFinder provided an overall ranking of the stability/suitability of these candidates. This study is the first to provide a comprehensive list of suitable reference genes for gene expression analyses in A. suturalis, which can serve to facilitate transcript expression study of related biological processes in this and related species.展开更多
The genus Adelphocoris(Hemiptera: Miridae) is a group of key insect pests in cotton fields in China that includes three dominant species: A. suturalis(Jakovlev), A. lineolatus(Goeze) and A. fasciaticollis(Reu...The genus Adelphocoris(Hemiptera: Miridae) is a group of key insect pests in cotton fields in China that includes three dominant species: A. suturalis(Jakovlev), A. lineolatus(Goeze) and A. fasciaticollis(Reuter). Previous field surveys have found that adults of these Adelphocoris species usually move onto specific host plants when the plants enter the flowering stage. To determine the potential trade-offs for this host-plant preference behavior, the performance of these three Adelphocoris spp. on flowering and non-flowering cotton and alfalfa were compared in the laboratory. The results showed that Adelphocoris spp. had significantly higher nymphal developmental and survival rates, along with increased adult longevity and fecundity on flowering cotton and alfalfa than on non-flowering plants of either species. In addition, compared with cotton plants, alfalfa generally promoted better performance of these three Adelphocoris species, especially for A. lineolatus. Simple correlation analysis indicated that female adult longevity was positively correlated to male adult longevity and female fecundity, and female fecundity was positively correlated to nymphal development and survival rates. This study demonstrated a positive correlation between adult preference and offspring/adult performance for all three Adelphocoris species, with no evidence of any trade-offs for this preference for flowering host plants. These findings support the hypothesis that hemimetabolous insects such as mirid bugs have positive adult preference-adult/nymphal performance relationships, which is likely due to the similar feeding habits and nutritional requirements of adults and nymphs.展开更多
基金funded by the National Special Key Project for Transgenic Breeding, China (2016ZX08011002)the Open Fundation of State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences (SKLOF201415)
文摘Quantitative reverse transcription polymerase chain reaction(qRT-PCR) is the most commonly-used tool for measurement of gene expression, but its accuracy and reliability depend on appropriate data normalization with the use of one or more stable reference genes. Adelphocoris suturalis is one of the most destructive pests of cotton, but until recently knowledge of its underlying molecular physiology had been hindered by a lack of molecular resources. To facilitate research on this pest, we evaluated 12 common housekeeping genes studied in insects(GAPDH, ACT, βACT, TBP, SDH, βTUB, EF1γ, EF1α, EF1δ, RPL32, RPS15, and RPL27) for their expression stability in A. suturalis when subjected to various experimental treatments, including three biotic(developmental stage and sex, tissue type, and metathoracic scent gland for varying developmental stages and sexes) and one abiotic(RNA interference injection) conditions. Four dedicated algorithms(ΔCt method, geNorm, BestKeeper and NormFinder) were used to analyze gene expression stability. In addition, RefFinder provided an overall ranking of the stability/suitability of these candidates. This study is the first to provide a comprehensive list of suitable reference genes for gene expression analyses in A. suturalis, which can serve to facilitate transcript expression study of related biological processes in this and related species.
基金supported by the Special Fund for Agro-Scientific Research in the Public Interest, China (201103012)the National Basic Research Program of China (2012CB114104)
文摘The genus Adelphocoris(Hemiptera: Miridae) is a group of key insect pests in cotton fields in China that includes three dominant species: A. suturalis(Jakovlev), A. lineolatus(Goeze) and A. fasciaticollis(Reuter). Previous field surveys have found that adults of these Adelphocoris species usually move onto specific host plants when the plants enter the flowering stage. To determine the potential trade-offs for this host-plant preference behavior, the performance of these three Adelphocoris spp. on flowering and non-flowering cotton and alfalfa were compared in the laboratory. The results showed that Adelphocoris spp. had significantly higher nymphal developmental and survival rates, along with increased adult longevity and fecundity on flowering cotton and alfalfa than on non-flowering plants of either species. In addition, compared with cotton plants, alfalfa generally promoted better performance of these three Adelphocoris species, especially for A. lineolatus. Simple correlation analysis indicated that female adult longevity was positively correlated to male adult longevity and female fecundity, and female fecundity was positively correlated to nymphal development and survival rates. This study demonstrated a positive correlation between adult preference and offspring/adult performance for all three Adelphocoris species, with no evidence of any trade-offs for this preference for flowering host plants. These findings support the hypothesis that hemimetabolous insects such as mirid bugs have positive adult preference-adult/nymphal performance relationships, which is likely due to the similar feeding habits and nutritional requirements of adults and nymphs.