Monitoring the population dynamics of a biocontrol agent in soil is important for understanding, predicting and increasing its efficacy. In this study, the population dynamics and the efficacy of a promising biocontrol agent against nematode, the fungus Hirsutella rhossiliensis, were investigated in greenhouse experiments with quantitative real-time polymerase chain reaction (PCR) and bioassay. To explore the effects of the fungus on nematode inoculum, soil infested with eggs or second-stage juveniles (J2) of Heterodera glycines was used. The results showed that the quantity of H. rhossiliensis DNA based on real-time PCR decreased over time, regardless of eggs or J2 as inoculum. The quantity of H. rhossiliensis DNA [femtogram (fg)/g soil] was highest (3.83 ± 1.96 × 108 fg/g soil) when the fungus was first added to soil and then decreased rapidly to 3.24 ± 1.22 × 107 fg/g in egg-infested soil and 7.34 ± 2.94 × 107 fg/g in J2-infested soil 17 days after planting, and then declined gradually between 17 and 59 days after planting. The data based on the bioassay showed that the percentage of J2 parasitized by H. rhossiliensis decreased throughout the experiment in both egg- and J2-infested soils. H. rhossiliensis controlled H. glycines more effectively in J2-infested soil than in egg-infested soil. At the end of the experiments (59 days after planting), nematode suppression and plant growth promotion in J2-infested soil were 79% and 55%, respectively, which were higher than the 34% and 2% in egg-infested soil. In soil that was not inoculated with the fungus, the number of H. glycines was 10 times higher in J2-infested soil than in egg-infested soil, indicating the greater nematode inoculum potential in soil infested with J2. DNA yield of H. rhossiliensis was statistically higher in J2-infested soil than in egg-infested soil in earlier period (prior to day 31) (p < 0.05), which is consistent with the hypothesis that the numbers of J2 present during and soon after fungal inoculation are critical for maintaining the population density and biocontrol efficiency of the fungus.
Bibliographical noteFunding Information:
This work was jointly supported by National Outstanding Youth Foundation (30625001), National 863 Plan of China (2006AA10A211) and Beijing Municipal Science and Technology Commission (D0706005040331 and D0705002040191). The authors also thank Prof. Bruce A. Jaffee (the University of California at Davis) for serving as pre-submission reviewers and for his valuable comments and suggestions.
- DNA yield
- Fungal parasitism
- Heterodera glycines
- Nematode suppression
- Real-time PCR