Growth inhibition of the cereal root pathogens Rhizoctonia solani AG8, R. oryzae and Gaeumannomyces graminis var. tritici by a recombinant 42-kDa endochitinase from Trichoderma harzianum

Yongchun Wu; Diter Von Wettstein; C. Gamini Kannangara; Jayaveeramuthu Nirmala; R. James Cook

doi:10.1080/09583150600699770

Growth inhibition of the cereal root pathogens Rhizoctonia solani AG8, R. oryzae and Gaeumannomyces graminis var. tritici by a recombinant 42-kDa endochitinase from Trichoderma harzianum

Yongchun Wu, Diter Von Wettstein, C. Gamini Kannangara, Jayaveeramuthu Nirmala, R. James Cook

Veterinary Population Medicine

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

12 Scopus citations

Abstract

The objective of this study was to determine the effectiveness of a 42-kDa endochitinase coded by the ThEn42 gene from Trichoderma harzianum as a potential source of transgenic resistance to Rhizoctonia root rot of barley caused by Rhizoctonia solani AG8 and/or R. oryzae . The gene cThEn42 was codon optimized (GC content increased from 53.3 to 65.1%) and then synthesized to produce the modified cThEn42GC in Pichia pastoris for in vitro tests. Two expression vectors were constructed: one with the fungal signal peptide and the fungal activation peptide [ FSP-FAP-cThEn(GC) ] and the other with barley chitinase 26 signal peptide followed by the fungal signal and activation peptides [ SP(HVChi26)-FSP-FAP-cThEn(GC) ]. N-terminal sequencing showed that, of two proteins secreted into liquid medium, FSP was cleaved off faithfully in one protein and both FSP and FAP were cleaved from the other protein. Purified endochitinase provided strong in vitro inhibition of both R. solani AG8 and R. oryzae . The enzyme had an intermediate inhibitory activity against Gaeumannomyces graminis var. tritici , and no inhibitory activity against Fusarium graminearum, F. pseudograminearum, and F. culmorum.

Original language	English (US)
Pages (from-to)	631-646
Number of pages	16
Journal	Biocontrol Science and Technology
Volume	16
Issue number	6
DOIs	https://doi.org/10.1080/09583150600699770
State	Published - Jun 1 2006

Bibliographical note

Funding Information:
We would like to thank Dr Norbert Wolf, Mr Rainer Stahl at Maltagen, Andernach, Germany, and Dr Henny Horvath for support with the synthesis of the endochitinase transgene. Financial support for this research was provided by the US Department of Agriculture, Agricultural Research Service, the Washington Barley Commission, the R.A. Nilan Distinguished Professorship and the R. J. Cook Endowed Chair in Wheat Research.

Keywords

Biological control
Endochitinase
Gaeumannomyces graminis
Pichia pastoris
R. oryzae
Rhizoctonia solani AG8
Soilborne plant pathogens
Trichoderma harzianum

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

Access

10.1080/09583150600699770

OpenUrl availability

Full text

Cite this

Growth inhibition of the cereal root pathogens Rhizoctonia solani AG8, R. oryzae and Gaeumannomyces graminis var. tritici by a recombinant 42-kDa endochitinase from Trichoderma harzianum. / Wu, Yongchun; Von Wettstein, Diter; Gamini Kannangara, C. et al.
In: Biocontrol Science and Technology, Vol. 16, No. 6, 01.06.2006, p. 631-646.

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

Wu, Y, Von Wettstein, D, Gamini Kannangara, C, Nirmala, J & James Cook, R 2006, 'Growth inhibition of the cereal root pathogens Rhizoctonia solani AG8, R. oryzae and Gaeumannomyces graminis var. tritici by a recombinant 42-kDa endochitinase from Trichoderma harzianum', Biocontrol Science and Technology, vol. 16, no. 6, pp. 631-646. https://doi.org/10.1080/09583150600699770

@article{8a0839cce62d4495a48983a96d04ba0a,

title = "Growth inhibition of the cereal root pathogens Rhizoctonia solani AG8, R. oryzae and Gaeumannomyces graminis var. tritici by a recombinant 42-kDa endochitinase from Trichoderma harzianum",

abstract = "The objective of this study was to determine the effectiveness of a 42-kDa endochitinase coded by the ThEn42 gene from Trichoderma harzianum as a potential source of transgenic resistance to Rhizoctonia root rot of barley caused by Rhizoctonia solani AG8 and/or R. oryzae . The gene cThEn42 was codon optimized (GC content increased from 53.3 to 65.1%) and then synthesized to produce the modified cThEn42GC in Pichia pastoris for in vitro tests. Two expression vectors were constructed: one with the fungal signal peptide and the fungal activation peptide [ FSP-FAP-cThEn(GC) ] and the other with barley chitinase 26 signal peptide followed by the fungal signal and activation peptides [ SP(HVChi26)-FSP-FAP-cThEn(GC) ]. N-terminal sequencing showed that, of two proteins secreted into liquid medium, FSP was cleaved off faithfully in one protein and both FSP and FAP were cleaved from the other protein. Purified endochitinase provided strong in vitro inhibition of both R. solani AG8 and R. oryzae . The enzyme had an intermediate inhibitory activity against Gaeumannomyces graminis var. tritici , and no inhibitory activity against Fusarium graminearum, F. pseudograminearum, and F. culmorum.",

keywords = "Biological control, Endochitinase, Gaeumannomyces graminis, Pichia pastoris, R. oryzae, Rhizoctonia solani AG8, Soilborne plant pathogens, Trichoderma harzianum",

author = "Yongchun Wu and {Von Wettstein}, Diter and {Gamini Kannangara}, C. and Jayaveeramuthu Nirmala and {James Cook}, R.",

note = "Funding Information: We would like to thank Dr Norbert Wolf, Mr Rainer Stahl at Maltagen, Andernach, Germany, and Dr Henny Horvath for support with the synthesis of the endochitinase transgene. Financial support for this research was provided by the US Department of Agriculture, Agricultural Research Service, the Washington Barley Commission, the R.A. Nilan Distinguished Professorship and the R. J. Cook Endowed Chair in Wheat Research.",

year = "2006",

month = jun,

day = "1",

doi = "10.1080/09583150600699770",

language = "English (US)",

volume = "16",

pages = "631--646",

journal = "Biocontrol Science and Technology",

issn = "0958-3157",

publisher = "Taylor and Francis Ltd.",

number = "6",

}

TY - JOUR

T1 - Growth inhibition of the cereal root pathogens Rhizoctonia solani AG8, R. oryzae and Gaeumannomyces graminis var. tritici by a recombinant 42-kDa endochitinase from Trichoderma harzianum

AU - Wu, Yongchun

AU - Von Wettstein, Diter

AU - Gamini Kannangara, C.

AU - Nirmala, Jayaveeramuthu

AU - James Cook, R.

N1 - Funding Information: We would like to thank Dr Norbert Wolf, Mr Rainer Stahl at Maltagen, Andernach, Germany, and Dr Henny Horvath for support with the synthesis of the endochitinase transgene. Financial support for this research was provided by the US Department of Agriculture, Agricultural Research Service, the Washington Barley Commission, the R.A. Nilan Distinguished Professorship and the R. J. Cook Endowed Chair in Wheat Research.

PY - 2006/6/1

Y1 - 2006/6/1

N2 - The objective of this study was to determine the effectiveness of a 42-kDa endochitinase coded by the ThEn42 gene from Trichoderma harzianum as a potential source of transgenic resistance to Rhizoctonia root rot of barley caused by Rhizoctonia solani AG8 and/or R. oryzae . The gene cThEn42 was codon optimized (GC content increased from 53.3 to 65.1%) and then synthesized to produce the modified cThEn42GC in Pichia pastoris for in vitro tests. Two expression vectors were constructed: one with the fungal signal peptide and the fungal activation peptide [ FSP-FAP-cThEn(GC) ] and the other with barley chitinase 26 signal peptide followed by the fungal signal and activation peptides [ SP(HVChi26)-FSP-FAP-cThEn(GC) ]. N-terminal sequencing showed that, of two proteins secreted into liquid medium, FSP was cleaved off faithfully in one protein and both FSP and FAP were cleaved from the other protein. Purified endochitinase provided strong in vitro inhibition of both R. solani AG8 and R. oryzae . The enzyme had an intermediate inhibitory activity against Gaeumannomyces graminis var. tritici , and no inhibitory activity against Fusarium graminearum, F. pseudograminearum, and F. culmorum.

AB - The objective of this study was to determine the effectiveness of a 42-kDa endochitinase coded by the ThEn42 gene from Trichoderma harzianum as a potential source of transgenic resistance to Rhizoctonia root rot of barley caused by Rhizoctonia solani AG8 and/or R. oryzae . The gene cThEn42 was codon optimized (GC content increased from 53.3 to 65.1%) and then synthesized to produce the modified cThEn42GC in Pichia pastoris for in vitro tests. Two expression vectors were constructed: one with the fungal signal peptide and the fungal activation peptide [ FSP-FAP-cThEn(GC) ] and the other with barley chitinase 26 signal peptide followed by the fungal signal and activation peptides [ SP(HVChi26)-FSP-FAP-cThEn(GC) ]. N-terminal sequencing showed that, of two proteins secreted into liquid medium, FSP was cleaved off faithfully in one protein and both FSP and FAP were cleaved from the other protein. Purified endochitinase provided strong in vitro inhibition of both R. solani AG8 and R. oryzae . The enzyme had an intermediate inhibitory activity against Gaeumannomyces graminis var. tritici , and no inhibitory activity against Fusarium graminearum, F. pseudograminearum, and F. culmorum.

KW - Biological control

KW - Endochitinase

KW - Gaeumannomyces graminis

KW - Pichia pastoris

KW - R. oryzae

KW - Rhizoctonia solani AG8

KW - Soilborne plant pathogens

KW - Trichoderma harzianum

UR - http://www.scopus.com/inward/record.url?scp=33745172466&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33745172466&partnerID=8YFLogxK

U2 - 10.1080/09583150600699770

DO - 10.1080/09583150600699770

M3 - Article

AN - SCOPUS:33745172466

SN - 0958-3157

VL - 16

SP - 631

EP - 646

JO - Biocontrol Science and Technology

JF - Biocontrol Science and Technology

IS - 6

ER -

Growth inhibition of the cereal root pathogens Rhizoctonia solani AG8, R. oryzae and Gaeumannomyces graminis var. tritici by a recombinant 42-kDa endochitinase from Trichoderma harzianum

Abstract

Bibliographical note

Keywords

UN SDGs

Access

OpenUrl availability

Other files and links

Fingerprint

Cite this