Wang Yunsheng, Zhou Lijuan, Yu Xiaoyue, Stover Ed, Luo Feng, Duan Yongping
College of Plant Protection, Hunan Agricultural UniversityChangsha, China; School of Computing, Clemson UniversityClemson, SC, USA.
U.S. Horticultural Research Laboratory, Agricultural Research Service Fort Pierce, FL, USA.
Front Plant Sci. 2016 Jun 28;7:933. doi: 10.3389/fpls.2016.00933. eCollection 2016.
Huanglongbing (HLB) is currently the most destructive disease of citrus worldwide. Although there is no immune cultivar, field tolerance to HLB within citrus and citrus relatives has been observed at the USDA Picos farm at Ft. Pierce, Florida, where plants have been exposed to a very high level of HLB pressure since 2006. In this study, we used RNA-Seq to evaluate expression differences between two closely related cultivars after HLB infection: HLB-tolerant "Jackson" grapefruit-like-hybrid trees and HLB susceptible "Marsh" grapefruit trees. A total of 686 genes were differentially expressed (DE) between the two cultivars. Among them, 247 genes were up-expressed and 439 were down-expressed in tolerant citrus trees. We also identified a total of 619 genes with significant differential expression of alternative splicing isoforms between HLB tolerant and HLB susceptible citrus trees. We analyzed the functional categories of DE genes using two methods, and revealed that multiple pathways have been suppressed or activated in the HLB tolerant citrus trees, which lead to the activation of the basal resistance or immunity of citrus plants. We have experimentally verified the expressions of 14 up-expressed genes and 19 down-expressed genes on HLB-tolerant "Jackson" trees and HLB-susceptible "Marsh" trees using real time PCR. The results showed that the expression of most genes were in agreement with the RNA-Seq results. This study provided new insights into HLB-tolerance and useful guidance for breeding HLB-tolerant citrus in the future.
黄龙病(HLB)是目前全球对柑橘最具毁灭性的病害。尽管不存在免疫品种,但在美国农业部位于佛罗里达州皮尔斯堡的皮科斯农场,已观察到柑橘及其近缘种对HLB具有田间耐受性,自2006年以来,该农场的植株一直处于极高的HLB压力之下。在本研究中,我们使用RNA测序技术评估了两种亲缘关系密切的品种在感染HLB后的表达差异:耐HLB的“杰克逊”葡萄柚类杂交树和感HLB的“马什”葡萄柚树。两个品种之间共有686个基因差异表达(DE)。其中,在耐病柑橘树中有247个基因上调表达,439个基因下调表达。我们还在耐HLB和感HLB的柑橘树之间共鉴定出619个具有显著差异表达的可变剪接异构体的基因。我们使用两种方法分析了DE基因的功能类别,结果表明耐HLB的柑橘树中多条途径被抑制或激活,这导致柑橘植株的基础抗性或免疫力被激活。我们使用实时PCR实验验证了耐HLB的“杰克逊”树和感HLB的“马什”树上14个上调表达基因和19个下调表达基因的表达情况。结果表明,大多数基因的表达与RNA测序结果一致。本研究为HLB耐受性提供了新的见解,并为未来培育耐HLB的柑橘提供了有用的指导。