Prevalence, incidence and molecular identification of root-knot nematodes of tomato in Pakistan

Abstract

Tomato is a widely grown vegetable in Pakistan. However, its production is severely constrained by root knot nematodes (RKNs). Accurate identification of RKNs is essential for an appropriate control program. The current study evaluated the prevalence, incidence and diversity of RKNs of tomato crops grown in the Khyber Pakhtunkhwa Province and their identification using molecular tools. A field survey, including 30 commercial tomato fields, was conducted in ten major tomato growing areas of Swat and Malakand divisions during spring 2010. The overall prevalence and incidence in the study area was 83.3 and 52.0%, respectively. Three species of RKNs, Meloidogyne arenaria, M. incognita and M. javanica were found alone or in mixed populations. Disease incidence ranged from 10% in Malakandher to 100% and 90 to 100% in Jabban and Malakand, respectively. The greatest galling index (GI) (5.0) and egg mass index (EMI) (5.0) was recorded in samples from Jabban, whereas the lowest GI and EMI were recorded in samples from Malakandher and Peshawar. The population density of RKNs was highest in roots (633.0 eggs and second-stage juveniles) and soil (533.0 eggs and second-stage juveniles) samples of Jabban. DNA amplification with rDNA (D2A-D3B) and (194 to 195) primers amplified 750 and 720 bp products for M. arenaria, M. incognita and M. javanica, respectively. Amplification with sequence characterized amplified regions (SCAR) primers produced characteristic products of 420 bp for M. arenaria (Far/Rar), 1200 bp for M. incognita (Finc/Rinc), and 670 bp for M. javanica (Fjav/Rjav). DNA amplification of mtDNA with C2F3/1108 primers yielded a 1700 bp size product for all three species of RKNs in comparison with 520 and 750 bp for M. chitwoodi and enterolobii, respectively, which were utilized as control. Sequencing the 28S rDNA product generated with the D2A-D3B primers did not differentiate among the three Meloidogyne spp. from the study area.

Key words: Meloidogyne, species identification, perineal pattern, sequence characterized amplified regions (SCAR) primers.