Hermsmeier D, Hart J K, Byzova M, Rodermel S R, Baum T J
Department of Plant Pathology, Iowa State University, Ames 50011, USA.
Mol Plant Microbe Interact. 2000 Mar;13(3):309-15. doi: 10.1094/MPMI.2000.13.3.309.
Gene expression changes in plant roots infected by plant-parasitic cyst nematodes are involved in the formation of nematode feeding sites. We analyzed mRNA abundance changes within roots of Arabidopsis thaliana during the early compatible interaction with Heterodera schachtii, the sugarbeet cyst nematode. Approximately 1,600 root sections, each containing a single parasitic nematode and its feeding site, and 1,600 adjacent, nematode-free root sections were excised from aseptic A. thaliana cultures 3 to 4 days after inoculation with H. schachtii. These tissue samples were termed infected and uninfected, respectively. Preparasitic nematodes were added to the uninfected tissue sample to maintain the nematode to plant tissue proportion. mRNA extracted from these two tissue samples was subjected to differential display analysis. Thirty-six cDNA clones corresponding to mRNA species with different abundance between both tissue samples were isolated. Of these clones, 24 were of A. thaliana origin and 12 were from H. schachtii. Differential display data predicted that the A. thaliana cDNA clones corresponded to 13 transcripts that were more abundant in the infected root sections and 11 transcripts that were more abundant in the uninfected root sections. H. schachtii cDNA clones were predicted to correspond to four transcripts that were more abundant in parasitic nematodes and to eight transcripts that were more abundant in preparasitic nematodes. In situ hybridization experiments confirmed the mRNA abundance changes in A. thaliana roots predicted by the differential display analyses for two A. thaliana clones.
植物寄生性孢囊线虫感染的植物根系中的基因表达变化参与了线虫取食位点的形成。我们分析了拟南芥根系在与甜菜孢囊线虫(Heterodera schachtii)早期亲和互作过程中的mRNA丰度变化。接种H. schachtii 3至4天后,从无菌培养的拟南芥中切下约1600个根段,每个根段包含一条寄生线虫及其取食位点,以及1600个相邻的、无线虫的根段。这些组织样本分别称为感染组和未感染组。将寄生前期线虫添加到未感染的组织样本中,以保持线虫与植物组织的比例。从这两个组织样本中提取的mRNA进行差异显示分析。分离出36个与两个组织样本中丰度不同的mRNA种类相对应的cDNA克隆。其中,24个克隆来源于拟南芥,12个克隆来自H. schachtii。差异显示数据预测,拟南芥cDNA克隆对应于13个在感染根段中丰度更高的转录本和11个在未感染根段中丰度更高的转录本。H. schachtii cDNA克隆预计对应于4个在寄生线虫中丰度更高的转录本和8个在寄生前期线虫中丰度更高的转录本。原位杂交实验证实了差异显示分析预测的两个拟南芥克隆在拟南芥根中的mRNA丰度变化。