Plant Pathology and Plant-Microbe Biology Section, School of Integrative Plant Science, Cornell University, Ithaca, NY 14853, USA.
Boyce Thompson Institute, Ithaca, NY 14853, USA.
Gigascience. 2022 Apr 28;11. doi: 10.1093/gigascience/giac035.
Huanglongbing, a devastating disease of citrus, is caused by the obligate, intracellular bacterium "Candidatus Liberibacter asiaticus" (CLas). CLas is transmitted by Diaphorina citri, the Asian citrus psyllid. Development of transmission-blocking strategies to manage huanglongbing relies on knowledge of CLas and D. citri interactions at the molecular level. Prior transcriptome analyses of D. citri point to changes in psyllid biology due to CLas infection but have been hampered by incomplete versions of the D. citri genome, proper host plant controls, and/or a lack of a uniform data analysis approach. In this work, we present lessons learned from a quantitative transcriptome analysis of excised heads, salivary glands, midguts, and bacteriomes from CLas-positive and CLas-negative D. citri using the chromosomal length D. citri genome assembly.
Each organ had a unique transcriptome profile and response to CLas infection. Though most psyllids were infected with the bacterium, CLas-derived transcripts were not detected in all organs. By analyzing the midgut dataset using both the Diaci_v1.1 and v3.0 D. citri genomes, we showed that improved genome assembly led to significant and quantifiable differences in RNA-sequencing data interpretation.
Our results support the hypothesis that future transcriptome studies on circulative, vector-borne pathogens should be conducted at the tissue-specific level using complete, chromosomal-length genome assemblies for the most accurate understanding of pathogen-induced changes in vector gene expression.
黄龙病是一种毁灭性的柑橘病害,由专性细胞内细菌“亚洲韧皮杆菌”(CLas)引起。CLas 通过柑橘木虱(Diaphorina citri)传播。开发阻断传播的策略来管理黄龙病依赖于对 CLas 和 D. citri 在分子水平上相互作用的了解。先前对柑橘木虱的转录组分析表明,由于 CLas 感染,木虱生物学发生了变化,但由于 D. citri 基因组的不完整版本、适当的宿主植物对照和/或缺乏统一的数据分析方法,这些分析受到了阻碍。在这项工作中,我们从使用 D. citri 染色体长度基因组组装体对来自 CLas 阳性和 CLas 阴性 D. citri 的切除头部、唾液腺、中肠和细菌体进行的定量转录组分析中总结了经验教训。
每个器官都有独特的转录组特征和对 CLas 感染的反应。尽管大多数木虱都被细菌感染,但并非所有器官都检测到 CLas 衍生的转录本。通过使用 Diaci_v1.1 和 v3.0 D. citri 基因组分析中肠数据集,我们表明,基因组组装的改进导致 RNA-seq 数据解释的显著和可量化差异。
我们的结果支持这样的假设,即未来对循环、媒介传播病原体的转录组研究应该在组织特异性水平上进行,使用完整的染色体长度基因组组装,以最准确地了解病原体诱导的媒介基因表达变化。