Comparison of expression profiles for Nicotiana benthamiana agroinfiltrated with caryophyllene synthase and SNARE-RNAi

Here we show for the first time that vesicle fusion is one of multiple pathways in plant cells involved in terpene emissions. We demonstrate a positive feedback on terpene production when vesicle fusion is inhibited in cells producing high levels of terpenes, which relates to an induced proteasome malfunction. N. benthamiana leaves were infiltrated with Agrobacterium containing the following expression constructs: (1) pBin+DsRed-RNAi, (2) MtVAMP721e-RNAi+DsRed-RNAi, (3) CST+DsRed-RNAi, (4) CST+MtVAMP721e-RNAi (six plants per construct, one leaf per plant). Infiltrated leaves were harvested seven days post agroinfiltration. Per sample two leaves were pooled, resulting in three biological replicates for each treatment. Followed by deep sequencing, using Illumina Hiseq 2000.

本研究首次证实，囊泡融合是植物细胞中参与萜烯排放的多条通路之一。本研究还发现，在高萜烯产量的植物细胞中抑制囊泡融合时，会对萜烯生成产生正反馈调控，该现象与诱导型蛋白酶体功能异常相关。将携带以下表达载体的农杆菌（Agrobacterium）浸润本氏烟（N. benthamiana）叶片：(1) pBin+DsRed-RNAi；(2) MtVAMP721e-RNAi+DsRed-RNAi；(3) CST+DsRed-RNAi；(4) CST+MtVAMP721e-RNAi（每株构建组对应6株植株，每株接种1片叶片）。农杆菌浸润后第7天收取被浸润的叶片，每份样本混合2片叶片，每组处理得到3个生物学重复，随后采用Illumina Hiseq 2000平台进行深度测序。