A screening technique to evaluate pigeonpea for resistance to Rotylenchulus reniformis*

Rotylenchulus reniformis is one of the most important nematode pests of pigeonpea. A simple greenhouse technique has been developed to aid evaluation of pigeonpea genotypes for resistance to R. reniformis. In greenhouse pot experiments, eggsacs of R. reniformis in pigeonpea (cv. ICPL 87) roots were counted by eye and with the aid of a stereoscopic microscope at 15, 30 and 45 days after seedling emergence in soils infested with various numbers of vermiform R. reniformis. Seedlings were rated for the number of eggsacs per root system on a one (no eggsacs) to nine (more than 50 eggsacs) scale. Eggsac ratings were more uniform when roots were evaluated at 30 – 45 days than at 15 days and an inoculum of 15 to 30 individuals/cm³ soil also helped reduce variability. Eggsacs were not easily visible without the aid of a stereoscopic microscope. Of the 14 stains tested, exposure of nematode-infected roots to 0.25% trypan blue for three min was effective in staining the eggsacs blue without staining the roots. Using the stain, the assessment of infestation by R. reniformis was equally accurate with or without the aid of a stereoscopic microscope. Exposure of eggsacs to trypan blue enhanced the emergence of juveniles from the eggsacs.