UDP-Glycosyltransferase Genes in the Striped Rice Stem Borer, Chilo suppressalis (Walker), and Their Contribution to Chlorantraniliprole Resistance
Abstract
:1. Introduction
2. Results
2.1. Chlorantraniliprole Toxicity and Synergists Assessment
2.2. Identification and Classification of UGT Genes
2.3. Screening of Overexpressed UGTs in C. suppressalis with Chlorantraniliprole Resistance
2.4. Expression Patterns of Overexpressed UGTs in Different Tissues
2.5. Confirmation of Overexpressed UGT Genes Function
3. Discussion
4. Materials and Methods
4.1. Insects and Sample Preparation
4.2. Toxicity and Synergism Bioassays
4.3. RNA Isolation and cDNA Synthesis
4.4. Identification of UGT Genes and Phylogenetic Analysis
4.5. Expression Patterns of UGT Genes in Resistant and Susceptible Strains
4.6. Functional Analysis of Overexpressed UGT Genes with RNAi
4.7. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
UGT | Uridine diphosphate glycosyltransferase |
UDP | Uridine diphosphate |
RyR | Ryanodine receptor |
RNAi | RNA interference |
dsRNA | Double-stranded RNA |
5-NU | 5-nitrouracil |
SUL | Sulfinpyrazone |
RR | Resistance ratio |
SR | Synergism ratio |
CL | Confidence limits |
ORF | Open reading frame |
MW | Molecular weight |
PI | Isoelectric point |
SP | Signal peptide |
EGFP | Enhanced green fluorescent protein |
qRT-PCR | Real-time quantitative reverse transcription polymerase chain reaction |
RT-PCR | Reverse transcription polymerase chain reaction |
PBS | Phosphate-buffered saline |
SD | Standard deviation |
ANOVA | Analysis of variance |
Tukey’s HSD | Tukey’s honest significant difference |
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Strain | Treatment | LC50 (95% CL) (mg·L−1) | Slope ± SE | x2 | RR | SR |
---|---|---|---|---|---|---|
YYS | Chlorantraniliprole | 12.76 (11.08–14.76) | 2.72 ± 0.31 | 5.96 | 1.00 | 1.00 |
Chlorantraniliprole + 5-NU | 10.48 (8.95–12.27) | 2.42 ± 0.35 | 5.17 | - | 1.22 | |
Chlorantraniliprole + SUL | 11.27 (9.76–13.03) | 2.70 ± 0.32 | 7.72 | - | 1.13 | |
YYR | Chlorantraniliprole | 565.48 (510.42–626.47) | 3.67 ± 0.22 | 7.31 | 44.32 | 1.00 |
Chlorantraniliprole + 5-NU | 167.47 (146.78–191.08) | 2.97 ± 0.29 | 2.68 | - | 3.38 * | |
Chlorantraniliprole + SUL | 158.71 (139.32–180.79) | 3.08 ± 0.28 | 7.75 | - | 3.56 * |
Gene Name | Accession Number | ORF Size (aa) | PI/MW (KDa) | SP | ‘UDPGT’ Domain | Best Blastx Match | ||
---|---|---|---|---|---|---|---|---|
Species Name 1 | GenBank Accession Number | Identity | ||||||
CsUGT33AF1 | MK135471 | 518 | 7.77/59.32 | 1–19 | 22–517 | Ha | XP_021197637.1 | 62% |
CsUGT33AG1 | MK135472 | 524 | 6.91/60.06 | 1–22 | 24–517 | Pp | XP_013139422.1 | 61% |
CsUGT33AG2 | MK135473 | 520 | 6.77/59.27 | 1–20 | 24–517 | Ha | AEW43120.1 | 55% |
CsUGT33AG3 | MK135474 | 520 | 9.14/59.67 | 1–23 | 25–519 | Ha | AEW43118.1 | 57% |
CsUGT340D1 | MK135475 | 523 | 7.24/59.54 | 1–19 | 21–519 | Bm | NP_001243963.1 | 52% |
CsUGT34A8 | MK135476 | 523 | 8.30/59.31 | – | 22–510 | At | XP_013184861.1 | 62% |
CsUGT39B10 | MK135477 | 528 | 9.00/61.43 | – | 110–517 | Bm | NP_001243980.1 | 69% |
CsUGT39B9 | MK135478 | 528 | 9.01/60.69 | 1–21 | 95–516 | Se | ANI21999.1 | 66% |
CsUGT40AJ1 | MK135479 | 514 | 8.87/58.20 | – | 19–513 | Ha | XP_021183292.1 | 56% |
CsUGT40AK1 | MK135480 | 519 | 8.39/59.76 | – | 29–518 | Ha | AEW43128.1 | 52% |
CsUGT40AK2 | MK135481 | 519 | 8.83/59.17 | – | 37–515 | Ha | AEW43128.1 | 52% |
CsUGT40AK3 | MK135482 | 520 | 8.76/59.33 | 1–18 | 27–516 | Ha | XP_021191114.1 | 54% |
CsUGT40AL1 | MK135483 | 531 | 9.32/61.01 | – | 32–531 | Ha | AEW43128.1 | 54% |
CsUGT40AM1 | MK135484 | 521 | 8.13/59.30 | 1–20 | 22–520 | Dp | OWR51610.1 | 59% |
CsUGT40AN1 | MK135485 | 515 | 9.00/57.36 | – | 22–514 | At | XP_013186135.1 | 56% |
CsUGT40AN2 | MK135486 | 514 | 8.94/57.42 | – | 22–513 | At | XP_013189977.1 | 56% |
CsUGT40AP1 | MK135487 | 519 | 9.22/58.68 | 1–19 | 24–518 | Bm | XP_012550238.1 | 56% |
CsUGT42C4 | MK135488 | 505 | 7.63/57.49 | 1–21 | 24–500 | Se | ANI22012.1 | 65% |
CsUGT42H1 | MK135489 | 524 | 8.67/59.39 | 1–22 | 24–523 | Pm | XP_014358643.1 | 61% |
CsUGT44A8 | MK135490 | 515 | 6.79/59.79 | – | 30–511 | Ha | XP_021186279.1 | 62% |
CsUGT45C1 | MK135491 | 515 | 8.33/58.49 | 1–19 | 24–502 | Pp | XP_013141171.1 | 54% |
CsUGT46A9 | MK135492 | 533 | 8.96/60.29 | 1–23 | 27–528 | At | XP_013183121.1 | 66% |
CsUGT47A7 | MK135493 | 530 | 8.94/60.44 | 1–18 | 19–510 | At | XP_013192144.1 | 78% |
CsUGT50A8 | MK135494 | 544 | 8.88/62.11 | 1–21 | 79–516 | Ha | XP_021184423.1 | 74% |
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Zhao, J.; Xu, L.; Sun, Y.; Song, P.; Han, Z. UDP-Glycosyltransferase Genes in the Striped Rice Stem Borer, Chilo suppressalis (Walker), and Their Contribution to Chlorantraniliprole Resistance. Int. J. Mol. Sci. 2019, 20, 1064. https://doi.org/10.3390/ijms20051064
Zhao J, Xu L, Sun Y, Song P, Han Z. UDP-Glycosyltransferase Genes in the Striped Rice Stem Borer, Chilo suppressalis (Walker), and Their Contribution to Chlorantraniliprole Resistance. International Journal of Molecular Sciences. 2019; 20(5):1064. https://doi.org/10.3390/ijms20051064
Chicago/Turabian StyleZhao, Jun, Lu Xu, Yang Sun, Pingping Song, and Zhaojun Han. 2019. "UDP-Glycosyltransferase Genes in the Striped Rice Stem Borer, Chilo suppressalis (Walker), and Their Contribution to Chlorantraniliprole Resistance" International Journal of Molecular Sciences 20, no. 5: 1064. https://doi.org/10.3390/ijms20051064