Molecular cloning and functional analysis of a necrosis and ethylene inducing protein (NEP) from Ganoderma boninense

Fungal necrosis and ethylene inducing proteins (NEPs) induce cell death and elicit strong immune responses in many crops. In this study, we report the cloning and characterization of a transcript encoding NEP from Ganoderma boninense which belongs to a family of fungi in Ganodermataceae that cause s...

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Main Authors: Teh, Chui Yao, Pang, Chin Lin, Tor, Xin Yen, Ho, Pei Yin, Lim, Yi Yi, Namasivayam, Parameswari, Ho, Chai Ling
Format: Article
Language:English
Published: Elsevier 2019
Online Access:http://psasir.upm.edu.my/id/eprint/81405/1/GANO.pdf
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author Teh, Chui Yao
Pang, Chin Lin
Tor, Xin Yen
Ho, Pei Yin
Lim, Yi Yi
Namasivayam, Parameswari
Ho, Chai Ling
author_facet Teh, Chui Yao
Pang, Chin Lin
Tor, Xin Yen
Ho, Pei Yin
Lim, Yi Yi
Namasivayam, Parameswari
Ho, Chai Ling
author_sort Teh, Chui Yao
collection UPM
description Fungal necrosis and ethylene inducing proteins (NEPs) induce cell death and elicit strong immune responses in many crops. In this study, we report the cloning and characterization of a transcript encoding NEP from Ganoderma boninense which belongs to a family of fungi in Ganodermataceae that cause serious infections of cacao, rubber, tea, coffee and palms. The open reading frame (ORF) encoding NEP inG. boninense(GbNEP) was cloned by 5′and 3’rapid amplification of cDNA ends (RACE) PCR. The transcript abundance of GbNEP increased in fungal culture treated with salicylic acid and jasmonic acid, respectively. The soluble recombinant Gb EP expressed in Escherichia coli BL21(DE3) pLysS was able to induce necrosis in tobacco and tomato but ineffective when applied to oil palm leaves and root tissues. The recombinant GbNEP could induce localized cell death, and production of hydrogen peroxide and superoxide in tobacco leaves. The addition of LaCl3(a calcium channel inhibitor) and EGTA (a Ca2+chelator) to tobacco leaves prior to the challenge with GbNEP significantly reduced the necrosis symptoms indicating that Ca2+is required for the action of GbNEP. In summary, this is the first report of NEP from Ganoderma species which may contribute to further research on the role of GbNEP during disease development.
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spelling upm.eprints-814052021-05-25T23:44:01Z http://psasir.upm.edu.my/id/eprint/81405/ Molecular cloning and functional analysis of a necrosis and ethylene inducing protein (NEP) from Ganoderma boninense Teh, Chui Yao Pang, Chin Lin Tor, Xin Yen Ho, Pei Yin Lim, Yi Yi Namasivayam, Parameswari Ho, Chai Ling Fungal necrosis and ethylene inducing proteins (NEPs) induce cell death and elicit strong immune responses in many crops. In this study, we report the cloning and characterization of a transcript encoding NEP from Ganoderma boninense which belongs to a family of fungi in Ganodermataceae that cause serious infections of cacao, rubber, tea, coffee and palms. The open reading frame (ORF) encoding NEP inG. boninense(GbNEP) was cloned by 5′and 3’rapid amplification of cDNA ends (RACE) PCR. The transcript abundance of GbNEP increased in fungal culture treated with salicylic acid and jasmonic acid, respectively. The soluble recombinant Gb EP expressed in Escherichia coli BL21(DE3) pLysS was able to induce necrosis in tobacco and tomato but ineffective when applied to oil palm leaves and root tissues. The recombinant GbNEP could induce localized cell death, and production of hydrogen peroxide and superoxide in tobacco leaves. The addition of LaCl3(a calcium channel inhibitor) and EGTA (a Ca2+chelator) to tobacco leaves prior to the challenge with GbNEP significantly reduced the necrosis symptoms indicating that Ca2+is required for the action of GbNEP. In summary, this is the first report of NEP from Ganoderma species which may contribute to further research on the role of GbNEP during disease development. Elsevier 2019-04 Article PeerReviewed text en http://psasir.upm.edu.my/id/eprint/81405/1/GANO.pdf Teh, Chui Yao and Pang, Chin Lin and Tor, Xin Yen and Ho, Pei Yin and Lim, Yi Yi and Namasivayam, Parameswari and Ho, Chai Ling (2019) Molecular cloning and functional analysis of a necrosis and ethylene inducing protein (NEP) from Ganoderma boninense. Physiological and Molecular Plant Pathology, 106. pp. 42-48. ISSN 0885-5765; ESSN: 1096-1178 https://www.sciencedirect.com/science/article/pii/S0885576518303096 10.1016/j.pmpp.2018.12.003
spellingShingle Teh, Chui Yao
Pang, Chin Lin
Tor, Xin Yen
Ho, Pei Yin
Lim, Yi Yi
Namasivayam, Parameswari
Ho, Chai Ling
Molecular cloning and functional analysis of a necrosis and ethylene inducing protein (NEP) from Ganoderma boninense
title Molecular cloning and functional analysis of a necrosis and ethylene inducing protein (NEP) from Ganoderma boninense
title_full Molecular cloning and functional analysis of a necrosis and ethylene inducing protein (NEP) from Ganoderma boninense
title_fullStr Molecular cloning and functional analysis of a necrosis and ethylene inducing protein (NEP) from Ganoderma boninense
title_full_unstemmed Molecular cloning and functional analysis of a necrosis and ethylene inducing protein (NEP) from Ganoderma boninense
title_short Molecular cloning and functional analysis of a necrosis and ethylene inducing protein (NEP) from Ganoderma boninense
title_sort molecular cloning and functional analysis of a necrosis and ethylene inducing protein nep from ganoderma boninense
url http://psasir.upm.edu.my/id/eprint/81405/1/GANO.pdf
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