Wang Haichuan, Eyun Seong-Il, Arora Kanika, Tan Sek Yee, Gandra Premchand, Moriyama Etsuko, Khajuria Chitvan, Jurzenski Jessica, Li Huarong, Donahue Maia, Narva Ken, Siegfried Blair
Department of Agronomy and Horticulture, University of Nebraska-Lincoln, Lincoln, NE 68583-0915, USA.
Center for Biotechnology, School of Biological Sciences, UNL, Lincoln, NE 68583, USA.
Toxins (Basel). 2017 Mar 30;9(4):124. doi: 10.3390/toxins9040124.
With Next Generation Sequencing technologies, high-throughput RNA sequencing (RNAseq) was conducted to examine gene expression in neonates of (LeConte) (Western Corn Rootworm, WCR) challenged with individual proteins of the binary insecticidal proteins, Cry34Ab1 and Cry35Ab1, and the combination of Cry34/Cry35Ab1, which together are active against rootworm larvae. Integrated results of three different statistical comparisons identified 114 and 1300 differentially expressed transcripts (DETs) in the Cry34Ab1 and Cry34/35Ab1 treatment, respectively, as compared to the control. No DETs were identified in the Cry35Ab1 treatment. Putative Bt binding receptors previously identified in other insect species were not identified in DETs in this study. The majority of DETs (75% with Cry34Ab1 and 68.3% with Cry34/35Ab1 treatments) had no significant hits in the NCBI nr database. In addition, 92 DETs were shared between Cry34Ab1 and Cry34/35Ab1 treatments. Further analysis revealed that the most abundant DETs in both Cry34Ab1 and Cry34/35Ab1 treatments were associated with binding and catalytic activity. Results from this study confirmed the nature of these binary toxins against WCR larvae and provide a fundamental profile of expression pattern of genes in response to challenge of the Cry34/35Ab1 toxin, which may provide insight into potential resistance mechanisms.
利用新一代测序技术，进行了高通量RNA测序（RNAseq），以检测受二元杀虫蛋白Cry34Ab1和Cry35Ab1的单个蛋白以及Cry34/Cry35Ab1组合（它们共同对根虫幼虫具有活性）攻击的西方玉米根虫（WCR）（LeConte）新生幼虫中的基因表达。三种不同统计比较的综合结果表明，与对照相比，Cry34Ab1处理和Cry34/35Ab1处理分别有114个和1300个差异表达转录本（DETs）。在Cry35Ab1处理中未鉴定出DETs。本研究在DETs中未鉴定出先前在其他昆虫物种中鉴定出的假定Bt结合受体。大多数DETs（Cry34Ab1处理中有75%，Cry34/35Ab1处理中有68.3%）在NCBI nr数据库中没有显著匹配。此外，Cry34Ab1处理和Cry34/35Ab1处理之间共有92个DETs。进一步分析表明，Cry34Ab1和Cry34/35Ab1处理中最丰富的DETs与结合和催化活性相关。本研究结果证实了这些二元毒素对WCR幼虫的作用性质，并提供了响应Cry34/35Ab1毒素攻击的基因表达模式的基本概况，这可能有助于深入了解潜在的抗性机制。
