An insight into the expression profile of defence-related genes in compatible and incompatible Oryza sativa-Meloidogyne graminicola interaction

A genome-wide association study in Indian wild rice accessions for resistance to the root-knot nematode Meloidogyne graminicola

“…Seeds of different rice accessions were surface-sterilized via 70% ethanol prior to soaking overnight in distilled water. Seeds were germinated in Petri dish containing wet filter paper in a growth chamber at 28 ± 2˚C and 4-5 days old seedlings were used for infection bioassays in Pluronic gel medium (PF-127, Sigma-Aldrich) as described previously [26,27]. Briefly, 60 ml of 23% PF-127 was poured into 150 × 20 mm Petri dish containing 7-9 uniformly distributed seedlings of identical genotype at <15˚C and approximately 30 RRKN J2s were inoculated at the root tip of each seedling by a pipette tip.…”

“…In order to avoid the secondary galling by RRKN, the experiment was terminated at 16 dpi as M. graminicola completes its life cycle in 19 days at 24-30˚C in well-drained soil [3,5,6]. In PF-127 medium, M. graminicola completes its life cycle in 15 days [26,27]. Plants were carefully harvested, roots were washed in water, galls and other parameters were observed under the microscope.…”

“…cDNA matching actin gene of O. sativa was amplified as a reference for constitutive expression using primers listed in S2 Table. Reaction mixture (10 μl) for each sample consisted of 5 μl SYBR Green PCR Master mix (Eurogentec), 750 nM of each primer and 1.5 ng cDNA. RT-qPCR cycling conditions and melt curve program were followed as previously described [26,27]. At least three biological and three technical replicates were used for each sample.…”

A genome-wide association study in Indian wild rice accessions for resistance to the root-knot nematode Meloidogyne graminicola

“…Seeds of different rice accessions were surface-sterilized via 70% ethanol prior to soaking overnight in distilled water. Seeds were germinated in Petri dish containing wet lter paper in a growth chamber at 28 ± 2 °C and 4-5 days old seedlings were used for infection bioassays in Pluronic gel medium (PF-127, Sigma-Aldrich) as described previously [36,37] Plants were regularly watered to eld capacity and fertilized with Yoshida's nutrient solution [22]. In order to avoid the secondary galling by RRKN, the experiment was terminated at 16 dpi as M. graminicola completes its life cycle in 19 days at 24-30 °C in well-drained soil [3,5,6].…”

“…Reaction mixture (10 µl) for each sample consisted of 5 µl SYBR Green PCR Master mix (Eurogentec), 750 nM of each primer and 1.5 ng cDNA. qRT-PCR cycling conditions and melt curve program were followed as previously described [36,37]. At least three biological and three technical replicates were used for each sample.…”