| 吴 羽，王一明，朱雁飞，高 俏，邱 林，李有志,,重金属、农药和植物病毒胁迫下白背飞虱内参基因的筛选[J].环境昆虫学报,(): |
| 重金属、农药和植物病毒胁迫下白背飞虱内参基因的筛选 |
| Screening of reference genes in the white-backed planthopper, Sogatella furcifera (Hemiptera: Delphacidae) under heavy metal, pesticide, and plant virus stress |
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| DOI： |
| 中文关键词: 白背飞虱 重金属镉 Flupyrimin Dinotefuran 南方水稻黑条矮缩病毒 内参基因 |
| 英文关键词:Sogatella furcifera cadmium flupyrimin dinotefuran southern rice black stripe dwarf virus reference gene |
| 基金项目: |
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| 摘要点击次数: 10 |
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| 中文摘要: |
| 本研究旨在筛选白背飞虱Sogatella furcifera在重金属、农药及其传播的南方水稻黑条矮缩病毒（SRBSDV）胁迫下稳定表达的内参基因，使白背飞虱在上述逆境下的基因表达量分析更加趋于标准化。采用qRT-PCR技术，通过GeNorm、Normfinder、BestKeeper、Delta Ct和RefFinder等软件分析了白背飞虱8个内参基因（EF1-α、RPS18、ACT、TUB、GAPDH、RPL9、RPL10、18S）在重金属镉胁迫、Flupyrimin和Dinotefuran药剂胁迫及SRBSDV胁迫下的表达稳定性。结果发现在不同胁迫下8个候选内参基因引物扩增特异性良好，重金属镉和Flupyrimin胁迫下最稳定的内参基因为RPL9和RPL10；Dinotefuran胁迫下最稳定的内参基因为ACT和RPL9；SRBSDV胁迫下最稳定的内参基因为TUB和RPL9。利用gstd1和Vg1为靶标基因对筛选到的内参基因稳定性进行鉴定，结果显示组合内参基因标准误差较单一内参基因的标准误差低，为最佳归一标准。本研究为利用qRT-PCR技术分析白背飞虱在重金属镉胁迫、Flupyrimin和Dinotefuran药剂胁迫以及SRBSDV胁迫下相关基因的表达研究提供了稳定的内参基因，为后续基因功能的研究奠定了基础。 |
| 英文摘要: |
| The study aims to identify stable reference genes for Sogatella furcifera under the stress of heavy metals, pesticides, and the Southern rice black-streaked dwarf virus (SRBSDV) that it transmits, thereby standardizing gene expression analysis under these adverse conditions. Using qRT-PCR technology, we evaluated the expression stability of eight candidate reference genes (EF1-α, RPS18, ACT, TUB, GAPDH, RPL9, RPL10, 18S) under the stress of heavy metal cadmium(Cd), Flupyrimin, Dinotefuran, and SRBSDV through GeNorm, NormFinder, BestKeeper, Delta Ct, and RefFinder software. The results demonstrated that all primers for the eight candidate reference genes exhibited high amplification specificity across different stress conditions. Under Cd and Flupyrimin stress, RPL9 and RPL10 were the most stable reference genes; Dinotefuran stress, ACT and RPL9 showed the highest stability; and SRBSDV stress, TUB and RPL9 were the most stable. The stability of the selected reference genes was further validated using gstd1 and Vg1 as target genes. The results indicated that the combined use of multiple reference genes resulted in a lower standard error compared to single reference genes, providing the optimal normalization standard. This study provides a set of stable reference genes for qRT-PCR-based gene expression analysis of S. furcifera under the stress of Cd, Flupyrimin, Dinotefuran, and SRBSDV, laying a foundation for subsequent research on gene function. |
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