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Life cycle of Paracoccus marginatus. E, egg; N1, sex-undifferentiated first-instar nymphs; N2, second-instar nymphs; N3, third-instar nymphs; N4, fourth-instar nymphs; A, adults.
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Paracoccus marginatus (Hemiptera: Pseudococcidae), known as the papaya mealybug, could cause considerable yield loss of several plants. To date, there is no molecular-based study of P. marginatus. Fatty acyl-CoA reductases (FARs) are key enzymes involved in wax synthesis. In the present study, we cloned and characterized coding sequences (CDS) of t...
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... cycle of P. marginatus consists of four instars in the females and five in the males, which is N1 (sex-undifferentiated nymphs), male and female second-instar nymphs (N2♂ and N2♀, respectively), male and female third-instar nymphs (N3♂ and N3♀, respectively), male fourth-instar nymphs (N4♂), and male and female adults (A♂ and A♀), respectively (Fig. 1). Three biological replicates were collected for each active stage of P. marginatus, and the insect numbers for each replicate were listed as: 100 of N1, 30 of N2 (♂ or ♀), 30 of N3 (♂ or ♀), 30 of N4 (♂), and 30 of adults (♂ or ♀); in addition, male and female insects were separated. All samples were ground in liquid nitrogen and ...
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... CDS of PmFAR1 and PmFAR2 were 1,590 and 1,497 bp in length, encoding 529 and 498 amino acids, respectively (Supp Fig. S1 [online only]). In addition, both PmFAR1 and PmFAR2 were nonsecretory proteins since no signal peptide was found by SignalP 4.1. The result of conserved domain analysis showed that both PmFAR1 and PmFAR2 have a Rossmann folding and a FAR_C region (Fig. 5), which belonged to the FAR_C superfamily (Supp Fig. S2 [online only]). The ...
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... Transcritpomics analysis is needed to confirm the quantitative, real-time, reverse transcriptase polymerase chain reaction (RT-qPCR), which is always used to test gene expression levels in biological samples and tissues [18][19][20] . RT-qPCR was calculated based on reference genes, such as actin 21,22 , tubulin 22 , EF1 20,22 , GAPDH 21 , ubiquitin 23 , and others. Actin, GAPDH, tubulin, and ubiquitin are used commonly in analysing gene expression levels in plants [23][24][25][26] . ...
... Transcritpomics analysis is needed to confirm the quantitative, real-time, reverse transcriptase polymerase chain reaction (RT-qPCR), which is always used to test gene expression levels in biological samples and tissues [18][19][20] . RT-qPCR was calculated based on reference genes, such as actin 21,22 , tubulin 22 , EF1 20,22 , GAPDH 21 , ubiquitin 23 , and others. Actin, GAPDH, tubulin, and ubiquitin are used commonly in analysing gene expression levels in plants [23][24][25][26] . ...
... Reference genes, such as actin, tubulin, tef1, NADP, and ubiquitin, are widely used in different species 18,20,[22][23][24] . However, appropriate reference genes are not uniform across diverse species, and even in different treatments or tissues of the same species [18][19][20][21][22][23] . We also attempted to use actin as the reference gene for validation of the expression levels according to the transcriptomic data. ...
Vaccinium bracteatum Thunb. (VBT) is widely distributed in the mountainous areas in eastern and southern China. VBT leaves have great medical value and can be used to stain rice to produce “Wumifan”. Its fruits also contain rich nutrients. However, there has been limited attention to exploring the molecular content of VBT. Previously, we performed RNA-seq on three typical VBT fruits that were at various stages of ripening, although a reliable reference gene was lost in validation.In this study, we selected ten candidate reference genes based on previous studies and transcriptomics analyses. Subsequently, these genes were evaluated using a combination of methods, including geNorm, NormFinder, and Bestkeeper, with a comprehensive ranking assessment. As a result, we found that the actin2, NADH, and ADK genes have high reliability for analysing the expression levels of genes involved in fruit development. Furthermore, the transcript levels of 15 DEGs from transcriptomic analysis were assessed using NADH as a reference gene, and RT-qPCR data were highly consistent with the transcriptomic data. These results provide reliable reference genes for further studying gene expression, which will be beneficial for comprehensively exploring VBT.
... Reference genes, such as actin, tubulin, tef1, NADP, and ubiquitin, are widely used in different species [18,20,[22][23][24]. However, appropriate reference genes are not uniform across diverse species, and even in different treatments or tissues of the same species [18][19][20][21][22][23]. We also attempted to use actin as the reference gene for validation of the expression levels according to the transcriptomic data. ...
... The genes were analysed using GeNorm, NormFinder, BestKeeper. Thus, because the scores of a former reference gene that was evaluated by the three methods are not uniform, many researchers then selected genes with high scores in GeNorm and NormFinder, or selected genes using a combination of the three methods [18,20,21,23]. As a result, the best reference gene remained to be identified. ...
Background: Vaccinium bracteatum Thunb. (VBT) is widely distributed in the mountainous areas in eastern and southern China. VBT leaves have great medical value and can be used to stain rice to produce “Wumifan”. Its fruits also contain rich nutrients. However, there has been limited attention to exploring the molecular content of VBT. Previously, we performed RNA-seq on three typical VBT fruits that were at various stages of ripening, although a reliable reference gene was lost in validation.
Results: In this study, we selected ten candidate reference genes based on previous studies and transcriptomics analyses. Subsequently, these genes were evaluated using a combination of methods, including geNorm, NormFinder, and Bestkeeper, with a comprehensive ranking assessment. As a result, we found that the actin2, NADH, and ADK genes have high reliability for analysing the expression levels of genes involved in fruit development. Furthermore, the transcript levels of 15 DEGs from transcriptomic analysis were assessed using NADH as a reference gene, and RT-qPCR data were highly consistent with the transcriptomic data.
Conclusions: These results provide a reliable reference gene for further studying gene expression, which will be beneficial for comprehensively exploring VBT.
... The activities of antioxidant enzymes (SOD, CAT, POD and PPO) of P. marginatus were differently affected by inoculation on susceptible and resistant cassavas (Fig. 5). The results showed that signi cantly lower levels of SOD, CAT, POD and PPO activities were detected in two resistant cassava plants than in three susceptible cassava plants (P < 0.05), decreasing by 31 ...
... B, Developmental duration (egg to female adult) of P. marginatus (F1) feeding on 15 cassava cultivars. The life cycle of P. marginatus was described by our previous studies(Liang et al. 2021). C and D, Determination of the total number of eggs laid per female and hatchability of P. marginatus. ...
Dangerous Paracoccus marginatus papaya mealybugs cause considerable threats and challenges to cassava production and processing. The deployment of resistant cultivars offers effective, economical and eco-friendly management strategies for pest management. We utilized P. marginatus mortality, development and reproduction to evaluate the resistance of fifteen cassava cultivars and conducted physiological and biochemical analyses when P. marginatus was fed on two resistant cultivars (Myanmar, C1115) and three susceptible cultivars (BRA900, Bread, SC205). Significantly lower digestive (amylase, sucrase, lipase), detoxification (glutathione-S-transferase and carboxylesterase) and antioxidant, including superoxide dismutase (SOD), catalase (CAT), peroxidase (POD), and polyphenol oxidase (PPO), enzyme activities were observed in P. marginatus feeding on resistant cultivars compared to susceptible cultivars. For resistant cultivars, a significant reduction was found in nutritional components containing free amino acids, nitrogen, soluble sugars and the secondary metabolite malondialdehyde. Additionally, significantly higher enzymatic activity (SOD, CAT, POD and PPO) levels and secondary metabolite quantities (total phenol and tannins) were found in resistant cultivars induced by P. marginatus compared with susceptible ones. Additionally, RT-qPCR tests showed that the transcripts of ten genes involved in nutrition, secondary metabolites and antioxidant activities were consistent with their physiology and biochemistry changes. Thus, resistant cultivars prevented P. marginatus populations from suffering lower P. marginatus damage by elevating secondary metabolite contents and antioxidant activities, reducing nutrition levels and decreasing enzymatic activities. This study will be beneficial in determining the important indexes for developing standard regulations to evaluate P. marginatus-resistant cassavas, helping the development of effective strategies for pest management.
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