Project description:We have infected the model legume Medicago truncatula with Meloidogyne hapla and harvested tissue over a time course. Transcriptome sequencing was performed on each sample using the Illumina RNA-Seq method. [Longitudinal Experiment] RNA was isolated from M. hapla eggs and pre-penetration juveniles (J2) and also from a time course of M. truncatula infected with M. hapla J2 at five time points: 1, 2, 4, 5, and 7 days after inoculation (DAI). Roots (local) and shoots (global) tissues from infected and uninfected plants were sampled. Each sample was loaded on five lanes for sequencing. Collectively, 22 samples were generated in total from - M. hapla egg and J2 (two samples), - infected M. truncatula root 1-2-4-5-7 DAI (five samples), - infected M. truncatula shoot 1-2-4-5-7 DAI (five samples), - uninfected M. truncatula root 1-2-4-5-7 DAI (five samples), - and uninfected M. truncatula shoot 1-2-4-5-7 DAI (five samples). Thus, 110 fastq files (= 22 samples x 5 lanes). [Diunrnal Experiment] M. truncatula roots inoculated with M. hapla was sampled at 6 time-points: 22:30, 2:00, 5:00, 6:30, 14:00, and 21:00. Lighting was turned off at 22:00 and turned on at 06:00. Four biological replicates were taken for each time point, providing 24 samples in total. Thus, 24 fastq files (= 6 time points x 4 replicates).
Project description:CHARACTERIZING MICROBIAL COMMUNITIES ASSOCIATED WITH THE NORTHERN ROOT-KNOT NEMATODE (MELOIDOGYNE HAPLA) OCCURRENCE AND SOIL HEALTH
Project description:We examined the efficacy of a bacterium for biocontrol of the root-knot nematode (RKN) Meloidogyne hapla in carrot (Daucus carota subsp. sativus) and tomato (Solanum lycopersicum). Among 542 bacterial isolates from various soils and plants, the highest nematode mortality was observed for treatments with isolate C1-7, which was identified as Bacillus cereus based on cultural and morphological characteristics, the Biolog program, and 16S rRNA sequencing analyses. The population density and the nematicidal activity of B. cereus C1-7 remained high until the end of culture in brain heart infusion broth, suggesting that it may have sustainable biocontrol potential. In pot experiments, the biocontrol efficacy of B. cereus C1-7 was high, showing complete inhibition of root gall or egg mass formation by RKN in carrot and tomato plants, and subsequently reducing RKN damage and suppressing nematode population growth, respectively. Light microscopy of RKN-infected carrot root tissues treated with C1-7 showed reduced formation of gall cells and fully developed giant cells, while extensive gall cells and fully mature giant cells with prominent cell wall ingrowths formed in the untreated control plants infected with RKNs. These histopathological characteristics may be the result of residual or systemic biocontrol activity of the bacterium, which may coincide with the biocontrol efficacies of nematodes in pots. These results suggest that B. cereus C1-7 can be used as a biocontrol agent for M. hapla.