Figure 4.

Spatial and temporal expression analysis of Hscm1. (A) Localization of chorismate mutase (CM) gene expression using whole‐mount in situ hybridization on second‐stage pre‐parasitic juveniles (J2s). Expression of Heterodera schachtii CM genes was observed in both subventral and dorsal glands of hatched J2s (left). No signal was detected using a sense probe as control (right). (B) Reverse transcriptase‐polymerase chain reaction (RT‐PCR) showing the expression of Hscm1 in different life stages of H. schachtii. The actin gene (Hs‐act) was used as an internal control. The different stages were as follows: developing eggs (lane 1); pre‐parasitic juveniles soaked (lane 2) or not soaked (lane 3) in cabbage root exudate; parasitic juveniles 4 days (lane 4) and 7 days (lane 5) after inoculation; adult females (lane 6); adult males (lane 7). In the negative control (lane 8), water was used instead of cDNA as template. Hscm1 mRNA is present in pre‐parasitic and parasitic stages, but not in developing eggs and mature males.