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

Limei Zhang; Xingzhong Liu; Shuifang Zhu; Senyu Chen

doi:10.1016/j.biocontrol.2005.08.002

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

Limei Zhang, Xingzhong Liu, Shuifang Zhu, Senyu Chen

Southern Research and Outreach Center

Research output: Contribution to journal › Article › peer-review

38 Scopus citations

Abstract

Hirsutella rhossiliensis, a nematophagous fungus, has shown potential in biocontrol of plant-parasitic nematodes. Monitoring the population dynamics of a biocontrol agent in soil requires comprehensive techniques and is essential to understand how it works. Bioassay based on the fungal parasitism on the juveniles of soybean cyst nematode, Heterodera glycines, can be used to evaluate the activity of the fungus but fails to quantify fungal biomass in soil. A real-time polymerase chain reaction (PCR) assay was developed to quantify the fungal population density in soil. The assay detected as little as 100 fg of fungal genomic DNA and 40 conidia g^-1 soil, respectively. The parasitism bioassay and the real-time PCR assay were carried out to investigate the presence, abundance and activity of H. rhossiliensis in soil after application of different inoculum levels. Both of the percentage of assay nematodes parasitized by H. rhossiliensis based on the parasitism bioassay and the DNA yield of the fungus quantified by real-time PCR increased significantly with the increase of the inoculum levels. The DNA yield of the fungus was positively correlated with the percentage of assay nematodes parasitized by H. rhossiliensis. The combination of the two is useful for monitoring fungal biomass and activity in soil.

Original language	English (US)
Pages (from-to)	316-323
Number of pages	8
Journal	Biological Control
Volume	36
Issue number	3
DOIs	https://doi.org/10.1016/j.biocontrol.2005.08.002
State	Published - Mar 2006

Bibliographical note

Funding Information:
We thank Dr. Hongyun Chen for assistance in operating the real-time PCR instrument and for his suggestion in this study, Mr. Wenjun Zhao for advice in designing primers and probe for real-time PCR, Dr. Richard A. Humber for providing Hirsutella cultures, and Ms. Rui Ma for reviewing this paper. This work was funded by K.C. Wong Education Foundation, Hong Kong.

Keywords

Heterodera glycines
Hirsutella rhossiliensis
Nematode juvenile
Parasitism bioassay
Real-time PCR
Soybean cyst nematode

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10.1016/j.biocontrol.2005.08.002

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

abstract = "Hirsutella rhossiliensis, a nematophagous fungus, has shown potential in biocontrol of plant-parasitic nematodes. Monitoring the population dynamics of a biocontrol agent in soil requires comprehensive techniques and is essential to understand how it works. Bioassay based on the fungal parasitism on the juveniles of soybean cyst nematode, Heterodera glycines, can be used to evaluate the activity of the fungus but fails to quantify fungal biomass in soil. A real-time polymerase chain reaction (PCR) assay was developed to quantify the fungal population density in soil. The assay detected as little as 100 fg of fungal genomic DNA and 40 conidia g-1 soil, respectively. The parasitism bioassay and the real-time PCR assay were carried out to investigate the presence, abundance and activity of H. rhossiliensis in soil after application of different inoculum levels. Both of the percentage of assay nematodes parasitized by H. rhossiliensis based on the parasitism bioassay and the DNA yield of the fungus quantified by real-time PCR increased significantly with the increase of the inoculum levels. The DNA yield of the fungus was positively correlated with the percentage of assay nematodes parasitized by H. rhossiliensis. The combination of the two is useful for monitoring fungal biomass and activity in soil.",

keywords = "Heterodera glycines, Hirsutella rhossiliensis, Nematode juvenile, Parasitism bioassay, Real-time PCR, Soybean cyst nematode",

author = "Limei Zhang and Xingzhong Liu and Shuifang Zhu and Senyu Chen",

note = "Funding Information: We thank Dr. Hongyun Chen for assistance in operating the real-time PCR instrument and for his suggestion in this study, Mr. Wenjun Zhao for advice in designing primers and probe for real-time PCR, Dr. Richard A. Humber for providing Hirsutella cultures, and Ms. Rui Ma for reviewing this paper. This work was funded by K.C. Wong Education Foundation, Hong Kong.",

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AU - Zhu, Shuifang

AU - Chen, Senyu

N1 - Funding Information: We thank Dr. Hongyun Chen for assistance in operating the real-time PCR instrument and for his suggestion in this study, Mr. Wenjun Zhao for advice in designing primers and probe for real-time PCR, Dr. Richard A. Humber for providing Hirsutella cultures, and Ms. Rui Ma for reviewing this paper. This work was funded by K.C. Wong Education Foundation, Hong Kong.

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N2 - Hirsutella rhossiliensis, a nematophagous fungus, has shown potential in biocontrol of plant-parasitic nematodes. Monitoring the population dynamics of a biocontrol agent in soil requires comprehensive techniques and is essential to understand how it works. Bioassay based on the fungal parasitism on the juveniles of soybean cyst nematode, Heterodera glycines, can be used to evaluate the activity of the fungus but fails to quantify fungal biomass in soil. A real-time polymerase chain reaction (PCR) assay was developed to quantify the fungal population density in soil. The assay detected as little as 100 fg of fungal genomic DNA and 40 conidia g-1 soil, respectively. The parasitism bioassay and the real-time PCR assay were carried out to investigate the presence, abundance and activity of H. rhossiliensis in soil after application of different inoculum levels. Both of the percentage of assay nematodes parasitized by H. rhossiliensis based on the parasitism bioassay and the DNA yield of the fungus quantified by real-time PCR increased significantly with the increase of the inoculum levels. The DNA yield of the fungus was positively correlated with the percentage of assay nematodes parasitized by H. rhossiliensis. The combination of the two is useful for monitoring fungal biomass and activity in soil.

AB - Hirsutella rhossiliensis, a nematophagous fungus, has shown potential in biocontrol of plant-parasitic nematodes. Monitoring the population dynamics of a biocontrol agent in soil requires comprehensive techniques and is essential to understand how it works. Bioassay based on the fungal parasitism on the juveniles of soybean cyst nematode, Heterodera glycines, can be used to evaluate the activity of the fungus but fails to quantify fungal biomass in soil. A real-time polymerase chain reaction (PCR) assay was developed to quantify the fungal population density in soil. The assay detected as little as 100 fg of fungal genomic DNA and 40 conidia g-1 soil, respectively. The parasitism bioassay and the real-time PCR assay were carried out to investigate the presence, abundance and activity of H. rhossiliensis in soil after application of different inoculum levels. Both of the percentage of assay nematodes parasitized by H. rhossiliensis based on the parasitism bioassay and the DNA yield of the fungus quantified by real-time PCR increased significantly with the increase of the inoculum levels. The DNA yield of the fungus was positively correlated with the percentage of assay nematodes parasitized by H. rhossiliensis. The combination of the two is useful for monitoring fungal biomass and activity in soil.

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KW - Hirsutella rhossiliensis

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

Abstract

Bibliographical note

Keywords

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