Arabidopsis endo-1,4-beta-glucanases are involved in the formation of root syncytia induced by Heterodera schachtii.

Cyst nematodes induce root syncytia with specific features such as hypertrophy, increased metabolic activity and fusion with adjacent cells. Cell walls of the syncytia undergo massive changes such as thickening, local dissolution and formation of ingrowths. Cell wall degrading and modifying proteins are apparently involved in syncytium formation but detailed knowledge of this is still limited. Therefore, we studied the regulation and function of the entire Arabidopsis endo-1,4-beta-glucanase gene family in syncytia induced by Heterodera schachtii. Endo-1,4-beta-glucanases hydrolyze the 1,4-beta-glucosidic linkages between glucose residues. Using semi-quantitative and quantitative approaches we identified seven genes that are upregulated in syncytia. Two of these genes, coding for secreted AtCel2 and membrane-bound KOR3, are shoot-specific but show high expression in syncytia at different developmental stages. In silico analysis of the promoter regions of both genes compared with other genes with modified regulation in nematode feeding sites did not reveal specific cis-acting elements that could be related to specific transcription in syncytia. However, motifs responsive to sugar and different plant hormones were identified. Accordingly, treatments with sucrose, gibberellic acid and NAA induced upregulation of AtCel2, whereas ABA triggered downregulation of both AtCel2 and KOR3 in roots. As AtCel2 is related to degradation of the cell wall matrix, we analysed the hemicellulose content in syncytia. The measured values resembled the expression pattern of AtCel2. A distinctly reduced number of females developed in cel2 and kor3 T-DNA mutants, and we therefore conclude that endo-1,4-beta-glucanases play an important role in the formation and function of syncytia.