Detection of the nematophagous fungus Hirsutella rhossiliensis in soil by real-time PCR and parasitism bioassay

Zhang, Limei; Liu, Xingzhong; Zhu, Shuifang; Chen, Senyu

doi:10.1016/j.biocontrol.2005.08.002

Cited by 43 publications

(21 citation statements)

References 26 publications

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“…The spore suspension was passed through a sieve with 25-µm openings, and the spores in the filtrate were adjusted to a concentration of 2×10 6 /mL. A 5-mL aliquot of each spore suspension was added to 100 mL of liquid medium [19] and cultured on a rotary shaker at 150 r min 1 at 20°C for 7 d. The mycelia were collected by centrifugation at 5000 r min 1 and were blended by a tissue blender (DS-1, Shanghai Angni Instruments Meters Co., Ltd, China) at 3000 r min 1 for 30 s. The mycelia were collected in 400 mL of sterile water and centrifuged at 5000 r min 1 again to collect the slurry. For tube soil assay, the slurry must be fresh without being dry.…”

Section: Fungal and Scn Preparationmentioning

confidence: 99%

“…The final Q-PCR reaction mixture of volume 25 μL contained 0.5 μL 200 nmol L 1 of each primer, 2 μL of 2.5 nmol L 1 dNTP, 12.5 μL of 2× SYBR Green buffer, 2 μL of the DNA template, and 7.5 μL of ddH 2 O. The primer F419 (5′-GCGCA-GTAGCTCCCAGAG-3′) and a reverse primer R480 (5′-TTGTTTTACGGCGTGACCG-3′) were used for the amplification with the following thermal cycling parameters: a denaturation step at 95C for 5 min, followed by 40 cycles at 95C for 15 s and 60C for 1 min [19]. The program was conducted in Bio-Rad CFX96 TM Real-time system.…”

Section: Standard Curve Preparation For Q-pcrmentioning

confidence: 99%

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Functional response of the fungus Hirsutella rhossiliensis to the nematode, Heterodera glycines

Shu

Lai

Yang

et al. 2015

Sci. China Life Sci.

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Functional response is a key index in determining the population fluctuation in predation. However, the lack of operable research system limits the studies on functional response of fungal predators. Hirsutella rhossiliensis is a dominant parasite of the soybean cyst nematode, Heterodera glycines. In a soil microcosm bioassay, we determined fungal biomass at different days within 21 days after inoculation, and parasitism rate of H. glycines by the fungus was determined. The functional response of H. rhossiliensis to H. glycines was established and found to be Holling's type III, which was influenced by mycelial densities. Meanwhile, we conducted anti-fungal analysis of metabolic fractions extracted from H. rhossiliensis to explain the potential mechanism of the intraspecific competition illustrated by functional response. The result of anti-fungal experiments indicated that the fungal predators had more complicated interaction at population level than expected, which might be regulated by self-inhibition metabolite(s). This study was the first functional response study of fungal predators in microcosm. With the increasing recognition of emerging fungal threats to animal, plant, and ecosystem health, the methodologies and hypotheses proposed in this study might inspire further research in fungal ecology. fungal predator, predation index, self-inhibition composition, soybean cyst nematode Citation:

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Section: Fungal and Scn Preparationmentioning

confidence: 99%

Section: Standard Curve Preparation For Q-pcrmentioning

confidence: 99%

Functional response of the fungus Hirsutella rhossiliensis to the nematode, Heterodera glycines

Shu

Lai

Yang

et al. 2015

Sci. China Life Sci.

Self Cite

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“…Fungal inoculum (a mycelial slurry) was prepared by growing the fungus in a liquid medium (Zhang et al, 2006) on an orbital shaker at 150 rpm for 7 days at 25°C. The fungal colonies were collected and blended following the method of Liu and Chen (2005).…”

Section: Fungal and Nematode Inoculummentioning

confidence: 99%

“…Hirsutella rhossiliensis DNA in soil was quantified by real-time PCR with specific primers (F419 and R480) and probe (P442) as described previously (Zhang et al, 2006). The TaqMan TM fluorescent probe was labeled at the 5 0 end with the reporter dye FAM (6-carboxy-fluorescein) and at the 3 0 end with the quencher dye TAMRA (6-carboxy-tetramenthylrhodamine) (Takara Bio Inc. Otsu, Shiga, Japan).…”

Section: Real-time Pcr Quantification Of H Rhossiliensis In Soilmentioning

confidence: 99%

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Population dynamics and biocontrol efficacy of the nematophagous fungus Hirsutella rhossiliensis as affected by stage of the soybean cyst nematode

et al. 2008

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a b s t r a c tMonitoring 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 Â 10 8 fg/g soil) when the fungus was first added to soil and then decreased rapidly to 3.24 ± 1.22 Â 10 7 fg/g in egg-infested soil and 7.34 ± 2.94 Â 10 7 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.

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Molecular Detection in Integrated Pest and Disease Management

Finetti-Sialer

Rosso²

2007

General Concepts in Integrated Pest and Disease Management

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Detection of the nematophagous fungus Hirsutella rhossiliensis in soil by real-time PCR and parasitism bioassay

Cited by 43 publications

References 26 publications

Functional response of the fungus Hirsutella rhossiliensis to the nematode, Heterodera glycines

Functional response of the fungus Hirsutella rhossiliensis to the nematode, Heterodera glycines

Population dynamics and biocontrol efficacy of the nematophagous fungus Hirsutella rhossiliensis as affected by stage of the soybean cyst nematode

Molecular Detection in Integrated Pest and Disease Management

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