Transcriptome and Difference Analysis of Fenpropathrin Resistant Predatory Mite, Neoseiulus barkeri (Hughes)
Abstract
:1. Introduction
2. Results
2.1. Resistance Selection
2.2. RNA-Sequencing Analysis
2.3. Annotation of Predicted Proteins
2.4. Functional Annotation
2.5. Cloning of the N. barkeri Sodium Channel Gene and Verification of Point Mutations
2.6. Identification and Differential Expression Analysis of P450s and GSTs
3. Discussion
4. Materials and Methods
4.1. Colony Rearing and Maintenance
4.2. Resistance Selection
4.3. Toxicity Testing and Determining the LC Value
4.4. RNA Preparation, Library Construction, and Sequencing
4.5. Sequence Analysis, de Novo Assembly, and Identification of Interesting Genes
4.6. Cloning and Mutations Confirmation of Sodium Channel Gene
4.7. Differential Expression Analysis of P450s and GSTs
5. Conclusions
Supplementary Materials
Acknowledgments
Author Contributions
Conflicts of Interest
Abbreviations
COG | Cluster of Orthologous Groups |
GO | Gene Ontology |
GST | Glutathione S-transferase |
KEGG | Kyoto Encyclopedia of Genes and Genomes |
LC50 | Lethal concentration 50 |
ORF | Open reading frame |
nr | Nonredundant |
P450 | Cytochrome P450 |
qPCR | Quantitative PCR |
RT-PCR | Reverse transcription PCR |
VGSC | Voltage-gated sodium channel |
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Strains | Regression Equation | X2 | 95% Confidence Interval (mg/L) | R2 | Resistance Ratio |
---|---|---|---|---|---|
FS | Y = 3.2619 + 1.1186X | 2.0824 | 35.79 (28.64–44.73) | 0.996 | 1 |
FR | Y = 0.9489 + 0.9321X | 0.0574 | 22,190.91 (2853.80–172,554.97) | 0.998 | 619.96 |
Sequencing | |
---|---|
Total number of reads | 25,192,607 |
Total number of nucleotides (nt) | 5,087,856,659 |
GC percentage (%) | 50.85 |
Q20 percentage (%) | 100.00 |
N percentage (%) | 0.01 |
Number of contigs | 1,385,792 |
Mean length of contigs (bp) | 74.76 |
Number of transcripts | 50,462 |
Mean length of transcripts (bp) | 1494.58 |
N50 of transcripts (bp) | 3007 |
Number of unigenes | 34,211 |
Mean length of unigenes (bp) | 1016.52 |
N50 of unigenes (bp) | 2126 |
Total annotated unigenes | 15,987 |
Unigenes annotations against nr | 15,866 |
Unigenes annotations against Swiss-Prot | 10,486 |
Unigenes annotations against KEGG | 5445 |
Unigenes annotations against COG | 5673 |
Unigenes annotations against GO | 8707 |
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Cong, L.; Chen, F.; Yu, S.; Ding, L.; Yang, J.; Luo, R.; Tian, H.; Li, H.; Liu, H.; Ran, C. Transcriptome and Difference Analysis of Fenpropathrin Resistant Predatory Mite, Neoseiulus barkeri (Hughes). Int. J. Mol. Sci. 2016, 17, 704. https://doi.org/10.3390/ijms17060704
Cong L, Chen F, Yu S, Ding L, Yang J, Luo R, Tian H, Li H, Liu H, Ran C. Transcriptome and Difference Analysis of Fenpropathrin Resistant Predatory Mite, Neoseiulus barkeri (Hughes). International Journal of Molecular Sciences. 2016; 17(6):704. https://doi.org/10.3390/ijms17060704
Chicago/Turabian StyleCong, Lin, Fei Chen, Shijiang Yu, Lili Ding, Juan Yang, Ren Luo, Huixia Tian, Hongjun Li, Haoqiang Liu, and Chun Ran. 2016. "Transcriptome and Difference Analysis of Fenpropathrin Resistant Predatory Mite, Neoseiulus barkeri (Hughes)" International Journal of Molecular Sciences 17, no. 6: 704. https://doi.org/10.3390/ijms17060704