研究目的
To profile the adverse effects of an EB trunk-injection agent on PWN by analyzing differential transcripts from the nematode’s whole genome through next-generation high-throughput sequencing.
研究成果
The findings further our understanding of the effects of EB for managing the PWD and may help to improve the pesticide-use strategies for controlling B. xylophilus.
研究不足
The study depends on predictions based on homologs studied in the model nematode Caenorhabditis elegans due to a lack of previous studies on PWN gene functions.
1：Experimental Design and Method Selection
A whole-organism transcriptomic analysis was conducted to understand the adverse effects of EB on B. xylophilus. The study involved toxicity assays and real-time quantitative PCR to validate transcriptional changes.
2：Sample Selection and Data Sources
The experimental PWN strain NB-6 was isolated from diseased Pinus massoniana. PWN fed on Botrytis cinerea cultured on potato dextrose agar (PDA) plates. Synchronized PWN adult stages were collected for the study.
3：List of Experimental Equipment and Materials
Technical grade EB (95% purity) was obtained from Zhejiang Shenghua Biok Biology Co. Ltd. The Baermann funnel method was used to separate nematodes from PDA plates. Total RNA was extracted using the TRIzol Max Bacterial RNA Isolation Kit. RNA-seq libraries were prepared and sequenced on an Illumina HiSeq 2500 platform.
4：Experimental Procedures and Operational Workflow
PWN adults were treated with EB, and total RNA was extracted. RNA-seq libraries were constructed and sequenced. Differentially expressed genes (DEGs) were identified and annotated. Bioassays assessed the effects of EB on PWN fecundity, egg hatching rate, development rate, and thrashing frequency.
5：Data Analysis Methods
DEGs were identified using the edgeR package. Functional enrichment analysis was performed using the R package clusterProfiler. qPCR was used to validate the expression levels of selected DEGs.
独家科研数据包，助您复现前沿成果，加速创新突破
获取完整内容-
TRIzol Max Bacterial RNA Isolation Kit
Thermo Fisher Scientific
Used for total RNA extraction from samples.
-
NanoDrop2000
Thermo Fisher Scientific
Used to measure RNA concentration and purity.
-
Agilent 2100 BioAnalyzer
Agilent
Used to check RNA integrity.
-
Illumina HiSeq 2500 platform
Illumina
Used for sequencing RNA-seq libraries.
-
PrimeScript? RT reagent Kit with gDNA Eraser
TaKaRa
Used for reverse transcription of RNA into cDNA.
-
SYBR Premix Ex Taq? II
TaKaRa
Used for real-time quantitative PCR.
-
qTOWER 2.2
Analytik Jena
Real-time PCR thermal cycler used for qPCR analysis.
-
登录查看剩余5件设备及参数对照表
查看全部
