Screening and Validation of Stable Reference Genes for qRT-PCR Analysis in Epicauta gorhami (Coleoptera: Meloidae) DOI Creative Commons
Guofeng Yang,

Xuetao Yu,

Yan Zhang

и другие.

Insects, Год журнала: 2024, Номер 15(12), С. 942 - 942

Опубликована: Ноя. 29, 2024

Epicauta gorhami is a hypermetamorphic insect that mainly forage soybeans during the adult stage. However, lack of appropriate references hinders our studying gene function in E. gorhami. In this study, referring to five computational tools (Ct value, geNorm, NormFinder, BestKeeper and RefFinder), stability 10 housekeeping genes (GAPDH, ACT, RPL4, RPL27, α-TUB, RPS18, EF1α, RPS28, RPL13 SOD) was assessed by qRT-PCR under three different conditions (adult ages, tissues/organs temperatures). The findings suggested SOD RPS18 were most ideal for examine transcripts among diverse ages at various temperatures; pair RPS28 reliable assess expressions tissues. Finally, relative expression levels EgUAP computed after normalization with across As expected, abundant foregut, trachea antenna scarce midgut, hindgut epidermis. These will lay solid foundation analyzing

Язык: Английский

Validation and Evaluation of Reference Genes for Quantitative Real-Time PCR Analysis in Mythimna loreyi (Lepidoptera: Noctuidae) DOI Creative Commons
L. Wang,

Chaoxia Yang,

Qing‐Yu Liu

и другие.

Insects, Год журнала: 2024, Номер 15(3), С. 185 - 185

Опубликована: Март 10, 2024

Quantitative real-time PCR (qRT-PCR) is a widely applied technique for accurately assessing the expression of target genes. In practice, evaluation gene requires appropriate reference To screen reliable genes evaluating via qRT-PCR in Mythimna loreyi, notorious migratory pest across Asia, Africa, Europe, and Australia, we assessed stability 13 candidate M. loreyi using ΔCt method, BestKeeper, Normfinder, GeNorm, web-based comprehensive platform RefFinder. These include RPL10, RPL27, RPL32, RPS3, TATA−box, GAPDH, AK, Actin, EF, α−tubulin, SOD, 18S rRNA, FTZ−F1, which frequently employed Lepidoptera insects. Our findings revealed that performance depended on experimental conditions. Specifically, RPL27 RPL10 were most suitable changes developmental stages, tissues, adult ages. The optimal recommended specific experiment conditions, instance, EF RPS3 mating status, AK temperature treatments, FTZ−F1 larva diet, diet treatments. Additionally, profiles pheromone-binding protein 2 (MlorPBP2) glutathione S-transferase (MlorGST1) used to validate This study provides accurate normalization data, laying groundwork studying loreyi.

Язык: Английский

Процитировано

5

Characterization of the ligand-binding properties of odorant-binding protein 38 from Riptortus pedestris when interacting with soybean volatiles DOI Creative Commons

Jianglong Guo,

Panjing Liu, Xiaofang Zhang

и другие.

Frontiers in Physiology, Год журнала: 2025, Номер 15

Опубликована: Янв. 6, 2025

Riptortus pedestris (Fabricius) (Hemiptera: Alydidae) is a major soybean pest throughout East Asia that relies on its advanced olfactory system for the perception of plant-derived volatile compounds and aggregation pheromones conspecific host plant localization. Odorant binding proteins (OBPs) facilitate transport odorant across sensillum lymph within insect system, enabling their interaction with receptors (ORs). Real-time quantitative PCR (qRT-PCR) analyses, fluorescence-based competitive assays, molecular docking analyses were applied to assess expression ligand-binding properties OBP38 from R. peddestris. The qRT-PCR revealed high levels RpedOBP38 in antennae without any apparent sex bias, it was also highly expressed adult stage. Recombinant prepared by expressing E. coli BL21 (DE3) followed purification Ni-chelating affinity column. found bind most strongly trans-2-decenal (Ki = 7.440) trans-2-nonenal 10.973), β-pinene, (+) -4-terpineol, carvacrol, methyl salicylate, (-)-carvone. 3D structure contains six α-helices three interlocked disulfide bridges comprising stable hydrophobic pocket. In final series several polar (e.g., His 94, Glu97) nonpolar Leu29, Ile59) residues be involved RpedOBP38-ligand binding. These data support role volatiles derived plants, providing important insight into mechanisms govern recognition pedestris, thereby informing development ecologically friendly approaches managing infestations.

Язык: Английский

Процитировано

0

Evaluation and Validation of Reference Genes for Gene Expression Analysis Using qRT-PCR in the Sugarcane Stem Borer Chilo sacchariphagus (Lepidoptera: Pyralidae) DOI Creative Commons
Zhixiong Wang, Shang XianKun, Ji-Li Wei

и другие.

Insects, Год журнала: 2024, Номер 15(8), С. 594 - 594

Опубликована: Авг. 4, 2024

(Lepidoptera: Pyralidae) is an economically important sugarcane pest. Although numerous studies were conducted on the physiological responses in

Язык: Английский

Процитировано

3

Comprehensive Screening and Validation of Stable Internal Reference Genes for Accurate qRT-PCR Analysis in Holotrichia parallela under Diverse Biological Conditions and Environmental Stresses DOI Creative Commons
Zhongjun Gong, Jing Zhang, Qi Chen

и другие.

Insects, Год журнала: 2024, Номер 15(9), С. 661 - 661

Опубликована: Авг. 30, 2024

is among the world's most destructive pests. For accurate qPCR and gene expression studies, selection of stable appropriate reference genes crucial. However, a thorough evaluation potential for use in

Язык: Английский

Процитировано

1

Identification of a Soybean Volatile Attractive for Riptortus pedestris Using Reverse Chemical Ecology Approach DOI
Xiaotong Zhang,

Xuan-Pu Luan,

Jiahang Wei

и другие.

Journal of Agricultural and Food Chemistry, Год журнала: 2024, Номер unknown

Опубликована: Ноя. 27, 2024

The bean bug Riptortus pedestris is a major soybean pest and cause of the stay-green symptoms. However, molecular mechanisms underlying its olfaction-mediated host-seeking behavior remain unclear. In this study, we compared antennae transcriptomes starved nonstarved adult R. pedestris, identifying four differentially expressed odorant receptor (OR) genes. Among these, RpedOR13 showed strong response to host volatile 2-phenylethanol (2-PE) in Xenopus oocyte assays, while electroantennography behavioral tests confirmed 2-PE as an effective attractant. Next, phylogenetic analysis identified RpedOR72b paralog RpedOR13, with subsequent assays confirming specific 2-PE. Additionally, RNA interference experiments highlighted crucial role detecting Taken together, these findings provide new insights into highlight successful application reverse chemical ecology OR-based screening bioactive compounds.

Язык: Английский

Процитировано

0

Screening and Validation of Stable Reference Genes for qRT-PCR Analysis in Epicauta gorhami (Coleoptera: Meloidae) DOI Creative Commons
Guofeng Yang,

Xuetao Yu,

Yan Zhang

и другие.

Insects, Год журнала: 2024, Номер 15(12), С. 942 - 942

Опубликована: Ноя. 29, 2024

Epicauta gorhami is a hypermetamorphic insect that mainly forage soybeans during the adult stage. However, lack of appropriate references hinders our studying gene function in E. gorhami. In this study, referring to five computational tools (Ct value, geNorm, NormFinder, BestKeeper and RefFinder), stability 10 housekeeping genes (GAPDH, ACT, RPL4, RPL27, α-TUB, RPS18, EF1α, RPS28, RPL13 SOD) was assessed by qRT-PCR under three different conditions (adult ages, tissues/organs temperatures). The findings suggested SOD RPS18 were most ideal for examine transcripts among diverse ages at various temperatures; pair RPS28 reliable assess expressions tissues. Finally, relative expression levels EgUAP computed after normalization with across As expected, abundant foregut, trachea antenna scarce midgut, hindgut epidermis. These will lay solid foundation analyzing

Язык: Английский

Процитировано

0