Reference Genes for Expression Analysis Using RT-qPCR in Cnaphalocrocis medinalis (Lepidoptera: Pyralidae)

Zhao, Xiaoyu; Guo, Jiawen; Lu, Yanhui; Sun, Tianyi; Tian, Jun-Ce; Huang, Jianlei; Xu, Hongxing; Wang, Zhengliang; Lü, Zhongxian

doi:10.3390/insects13111046

Cited by 8 publications

(7 citation statements)

References 64 publications

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“…However, this topic has yet to get much attention, and its significance does not seem adequately understood. Because the option of reference genes varies drastically, insects of the same species, in terms of insect morphology, developmental stage, temperature, sex, and diet conditions, suggest that there is also no absolute generality between benchmark genes for the same species [ 29 , 30 , 31 , 32 ] . For instance, PGK and RPL13 are acceptable internal housekeeping genes in Cnaphalocrocis medinalis under different sexes [ 31 ].…”

Section: Discussionmentioning

confidence: 99%

“…Because the option of reference genes varies drastically, insects of the same species, in terms of insect morphology, developmental stage, temperature, sex, and diet conditions, suggest that there is also no absolute generality between benchmark genes for the same species [ 29 , 30 , 31 , 32 ] . For instance, PGK and RPL13 are acceptable internal housekeeping genes in Cnaphalocrocis medinalis under different sexes [ 31 ]. PGK and EF1α are stable housekeeping genes expressed in Cnaphalocrocis medinalis larvae under different temperature conditions [ 31 ].…”

Section: Discussionmentioning

confidence: 99%

“…For instance, PGK and RPL13 are acceptable internal housekeeping genes in Cnaphalocrocis medinalis under different sexes [ 31 ]. PGK and EF1α are stable housekeeping genes expressed in Cnaphalocrocis medinalis larvae under different temperature conditions [ 31 ]. Thus, for specific experimental treatments of L. invasa , it is indispensable to select appropriate housekeeping genes.…”

Section: Discussionmentioning

confidence: 99%

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Identification and Validation of Reference Genes for Expression Analysis Using RT-qPCR in Leptocybe invasa Fisher and La Salle (Hymenoptera: Eulophidae)

Liu

Zhou

Qiu

et al. 2023

Insects

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Leptocybe invasa (Hymenoptera: Eulophidae) is a globally intrusive pest. Despite extensive research into the physiological responses of this pest, our understanding of the molecular mechanisms still needs to be improved. We want to accurately investigate the expression of L. invasa’s target genes, so it is imperative to select fitting reference genes. In this study, eight housekeeping genes’ stability (RPS30, ACTR, 18S rRNA, ACT, RPL18, GAPDH, 28S rRNA, and TUB) was tested under five different experimental conditions, including male or female adults, somites (head, thorax, and abdomen), temperatures (0 °C, 25 °C, and 40 °C), diets (starvation, clear water, 10% honey water, Eucalyptus sap), and pesticides (acetone was used as a control, imidacloprid, monosultap). Gene stability was calculated using RefFinder, which integrates four algorithms (the ∆Ct method, geNorm, NormFinder, and BestKeeper). The findings implied that ACT and ACTR were the most accurate when comparing sexes. For analyzing different somites, 28S rRNA and RPL18 were ideal; the 28S rRNA and RRS30 were perfect for analyzing at different temperatures. The combination of ACT and GAPDH helped to analyze gene expression in different diets, and GAPDH and 28S rRNA were suitable for various pesticide conditions. Overall, this research offers a complete list of reference genes from L. invasa for precise analysis of target gene expression, which can improve the trustworthiness of RT-qPCR and lay the foundation for further investigations into the gene function of this pest.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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Identification and Validation of Reference Genes for Expression Analysis Using RT-qPCR in Leptocybe invasa Fisher and La Salle (Hymenoptera: Eulophidae)

Liu

Zhou

Qiu

et al. 2023

Insects

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“…Consistent with our findings, other studies have shown that RPS11 expression is stable across various developmental stages in Tuta absoluta but is the lowest in diverse adult tissues [ 41 ]. In Cnaphalocrocis medinalis , RPL13 emerges as the most stable ribosomal protein across different adult ages but becomes the most unstable one in larval tissues and in larvae under temperature treatments [ 42 ]. These investigations suggest that the expression of ribosomal proteins varies significantly under different experimental conditions.…”

Section: Discussionmentioning

confidence: 99%

Validation and Evaluation of Reference Genes for Quantitative Real-Time PCR Analysis in Mythimna loreyi (Lepidoptera: Noctuidae)

Wang,

Yang,

Liu

et al. 2024

Insects

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

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“…The seeds of three rice varieties, YY1540 (Ningbo Seed Co., Ltd, Ningbo, China), XS134, and ZZ39 (Zhejiang Wuwangnong Seeds Shareholding Co., Ltd, Hangzhou, China), were soaked for 24 h and drained, and then covered with moist gauze to induce germination for 48 h (Zhao et al 2022 ). They were planted in a white plastic bowl (26 cm × 17 cm × 8 cm in length, width, and height) to raise seedlings and transferred to flower pots (diameter 12 cm) after 12 d. All three varieties were planted in the greenhouse at Zhejiang Academy of Agricultural Sciences in Hangzhou, China (30.31° N, 120.20° E), and were used for experiments 40 d after transplantation.…”

Section: Methodsmentioning

confidence: 99%

Defense Responses of Different Rice Varieties Affect Growth Performance and Food Utilization of Cnaphalocrocis medinalis Larvae

Zhao,

Xu,

Yang

et al. 2024

Rice

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Rice leaf folder, Cnaphalocrocis medinalis (Guenée), is one of the most serious pests on rice. At present, chemical control is the main method for controlling this pest. However, the indiscriminate use of chemical insecticides has non-target effects and may cause environmental pollution. Besides, leaf curling behavior by C. medinalis may indirectly reduce the efficacy of chemical spray. Therefore, it is crucial to cultivate efficient rice varieties resistant to this pest. Previous studies have found that three different rice varieties, Zhongzao39 (ZZ39), Xiushui134 (XS134), and Yongyou1540 (YY1540), had varying degrees of infestation by C. medinalis. However, it is currently unclear whether the reason for this difference is related to the difference in defense ability of the three rice varieties against the infestation of C. medinalis. To explore this issue, the current study investigated the effects of three rice varieties on the growth performance and food utilization capability of the 4th instar C. medinalis. Further, it elucidated the differences in defense responses among different rice varieties based on the differences in leaf physiological and biochemical indicators and their impact on population occurrence. The results showed that the larval survival rate was the lowest, and the development period was significantly prolonged after feeding on YY1540. This was not related to the differences in leaf wax, pigments, and nutritional components among the three rice varieties nor to the feeding preferences of the larvae. The rate of superoxide anion production, hydrogen peroxide content, and the activity of three protective enzymes were negatively correlated with larval survival rate, and they all showed the highest in YY1540 leaves. Compared to other tested varieties, although the larvae feeding on YY1540 had higher conversion efficiency of ingested food and lower relative consumption rate, their relative growth was faster, indicating stronger food utilization capability. However, they had a lower accumulation of protein. This suggests that different rice varieties had different levels of oxidative stress after infestation by C. medinalis. The defense response of YY1540 was more intense, which was not conducive to the development of the larvae population. These results will provide new insights into the interaction mechanism between different rice varieties and C. medinalis and provide a theoretical basis for cultivating rice varieties resistant to this pest.

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Reference Genes for Expression Analysis Using RT-qPCR in Cnaphalocrocis medinalis (Lepidoptera: Pyralidae)

Cited by 8 publications

References 64 publications

Identification and Validation of Reference Genes for Expression Analysis Using RT-qPCR in Leptocybe invasa Fisher and La Salle (Hymenoptera: Eulophidae)

Identification and Validation of Reference Genes for Expression Analysis Using RT-qPCR in Leptocybe invasa Fisher and La Salle (Hymenoptera: Eulophidae)

Validation and Evaluation of Reference Genes for Quantitative Real-Time PCR Analysis in Mythimna loreyi (Lepidoptera: Noctuidae)

Defense Responses of Different Rice Varieties Affect Growth Performance and Food Utilization of Cnaphalocrocis medinalis Larvae

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