从边缘附球菌(半翅目:粉虱科)中分析两种脂肪酰基辅酶 A 还原酶基因转录模式的参考基因选择。
Reference Gene Selection for Analyzing the Transcription Patterns of Two Fatty Acyl-CoA Reductase Genes From Paracoccus marginatus (Hemiptera: Pseudococcidae).
机构信息
Environment and Plant Protection Institute, Chinese Academy of Tropical Agricultural Sciences/Key Laboratory of Integrated Pest Management on Tropical Crops, Ministry of Agriculture and Rural Affairs, Haikou, Hainan 571101, China.
Sanya Research Academy, Chinese Academy of Tropical Agriculture Science/Hainan Key Laboratory for Biosafety Monitoring and Molecular Breeding in Off-Season Reproduction Regions, Sanya, Hainan 572000, China.
出版信息
J Insect Sci. 2021 Sep 1;21(5). doi: 10.1093/jisesa/ieab072.
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 two FAR genes from P. marginatus. The results showed that PmFAR1 and PmFAR2 CDS were 1,590 and 1,497 bp in length, respectively, and sequence analysis indicated that these two genes both had the conservative motifs belonging to FAR_C superfamily. Furthermore, seven candidate reference genes were analyzed for their expression stability by using common algorithms including comparative ΔCq method, geNorm, NormFinder, BestKeeper, and RefFinder. Eventually, β-actin and GAPDH were the best reference genes in evaluating the expression of those two FAR genes. We found that PmFAR1 and PmFAR2 showed distinct expression patterns in different life stages. Moreover, the transcription of PmFAR1 and PmFAR2 in P. marginatus fed on resistant cassava cultivars was significantly lower compared with those fed on susceptible ones, indicating the potential function of FAR genes in cassava resistance to P. marginatus. The present study might help in better understanding the molecular mechanism of cassava resistance to mealybug.
边缘臀纹粉蚧(Hemiptera: Pseudococcidae),又名番木瓜粉蚧,可导致多种植物的产量严重损失。迄今为止,尚未有针对边缘臀纹粉蚧的基于分子的研究。脂肪酸酰基辅酶 A 还原酶(FARs)是参与蜡合成的关键酶。在本研究中,我们从边缘臀纹粉蚧中克隆并鉴定了两个 FAR 基因的编码序列(CDS)。结果表明,PmFAR1 和 PmFAR2 的 CDS 分别长 1590 和 1497bp,序列分析表明这两个基因都具有 FAR_C 超家族的保守基序。此外,使用比较 ΔCq 法、geNorm、NormFinder、BestKeeper 和 RefFinder 等常用算法分析了七个候选参考基因的表达稳定性。最终,β-肌动蛋白和 GAPDH 被选为评估这两个 FAR 基因表达的最佳参考基因。我们发现 PmFAR1 和 PmFAR2 在不同的生活阶段表现出不同的表达模式。此外,与食用感病品种相比,边缘臀纹粉蚧食用抗感品种时,PmFAR1 和 PmFAR2 的转录水平显著降低,表明 FAR 基因在木薯抗粉蚧中的潜在功能。本研究可能有助于更好地理解木薯对粉蚧抗性的分子机制。
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