Resistance to Aflatoxin Accumulation in Maize Mediated by Host-Induced Silencing of the Aspergillus flavus Alkaline Protease (alk) Gene

Aspergillus flavus is a fungal pathogen that infects maize and produces aflatoxins. Host-Induced Gene Silencing (HIGS) has been shown to reduce host infection by various fungal pathogens. Here, the A. flavus alkaline protease (alk) gene was ta...

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Autores principales:	Olanike Omolehin, Yenjit Raruang, Dongfang Hu, Zhu-Qiang Han, Qijian Wei, Kan Wang, Kanniah Rajasekaran, Jeffrey W. Cary, Zhi-Yuan Chen
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	<i>Aspergillus flavus</i> host induced gene silencing alkaline protease (<i>alk</i>) aflatoxin resistance transgenic maize small RNA Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/f3c34a0d00c94124b0e7291fa3af2aed
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spelling	oai:doaj.org-article:f3c34a0d00c94124b0e7291fa3af2aed2021-11-25T18:05:35ZResistance to Aflatoxin Accumulation in Maize Mediated by Host-Induced Silencing of the <i>Aspergillus flavus</i> Alkaline Protease (<i>alk</i>) Gene10.3390/jof71109042309-608Xhttps://doaj.org/article/f3c34a0d00c94124b0e7291fa3af2aed2021-10-01T00:00:00Zhttps://www.mdpi.com/2309-608X/7/11/904https://doaj.org/toc/2309-608X<i>Aspergillus flavus</i> is a fungal pathogen that infects maize and produces aflatoxins. Host-Induced Gene Silencing (HIGS) has been shown to reduce host infection by various fungal pathogens. Here, the <i>A. flavus</i> alkaline protease (<i>alk</i>) gene was targeted for silencing through HIGS. An RNAi vector carrying a portion of the <i>alk</i> gene was incorporated into the B104 maize genome. Four out of eight transformation events containing the <i>alk</i> gene, Alk-3, Alk-4, Alk-7 and Alk-9, were self-pollinated to T4/T6 generations. At T3, the Alk-transgenic lines showed up to 87% reduction in aflatoxin accumulation under laboratory conditions. T4 transgenic Alk-3 and Alk-7 lines, and T5 and T6 Alk-4 and Alk-9 showed an average of 84% reduction in aflatoxin accumulation compared to their null controls under field inoculations (<i>p</i> < 0.05). F<sub>1</sub> hybrids of three elite maize inbred lines and the transgenic lines also showed significant improvement in aflatoxin resistance (<i>p</i> < 0.006 to <i>p</i> < 0.045). Reduced <i>A. flavus</i> growth and levels of fungal <i>ß-tubulin</i> DNA were observed in transgenic kernels during in vitro inoculation. Alk-4 transgenic leaf and immature kernel tissues also contained about 1000-fold higher levels of <i>alk</i>-specific small RNAs compared to null controls, indicating that the enhanced aflatoxin resistance in the transgenic maize kernels is due to suppression of <i>A. flavus</i> infection through HIGS of <i>alk</i> gene.Olanike OmolehinYenjit RaruangDongfang HuZhu-Qiang HanQijian WeiKan WangKanniah RajasekaranJeffrey W. CaryZhi-Yuan ChenMDPI AGarticle<i>Aspergillus flavus</i>host induced gene silencingalkaline protease (<i>alk</i>)aflatoxin resistancetransgenic maizesmall RNABiology (General)QH301-705.5ENJournal of Fungi, Vol 7, Iss 904, p 904 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	<i>Aspergillus flavus</i> host induced gene silencing alkaline protease (<i>alk</i>) aflatoxin resistance transgenic maize small RNA Biology (General) QH301-705.5
spellingShingle	<i>Aspergillus flavus</i> host induced gene silencing alkaline protease (<i>alk</i>) aflatoxin resistance transgenic maize small RNA Biology (General) QH301-705.5 Olanike Omolehin Yenjit Raruang Dongfang Hu Zhu-Qiang Han Qijian Wei Kan Wang Kanniah Rajasekaran Jeffrey W. Cary Zhi-Yuan Chen Resistance to Aflatoxin Accumulation in Maize Mediated by Host-Induced Silencing of the <i>Aspergillus flavus</i> Alkaline Protease (<i>alk</i>) Gene
description	<i>Aspergillus flavus</i> is a fungal pathogen that infects maize and produces aflatoxins. Host-Induced Gene Silencing (HIGS) has been shown to reduce host infection by various fungal pathogens. Here, the <i>A. flavus</i> alkaline protease (<i>alk</i>) gene was targeted for silencing through HIGS. An RNAi vector carrying a portion of the <i>alk</i> gene was incorporated into the B104 maize genome. Four out of eight transformation events containing the <i>alk</i> gene, Alk-3, Alk-4, Alk-7 and Alk-9, were self-pollinated to T4/T6 generations. At T3, the Alk-transgenic lines showed up to 87% reduction in aflatoxin accumulation under laboratory conditions. T4 transgenic Alk-3 and Alk-7 lines, and T5 and T6 Alk-4 and Alk-9 showed an average of 84% reduction in aflatoxin accumulation compared to their null controls under field inoculations (<i>p</i> < 0.05). F<sub>1</sub> hybrids of three elite maize inbred lines and the transgenic lines also showed significant improvement in aflatoxin resistance (<i>p</i> < 0.006 to <i>p</i> < 0.045). Reduced <i>A. flavus</i> growth and levels of fungal <i>ß-tubulin</i> DNA were observed in transgenic kernels during in vitro inoculation. Alk-4 transgenic leaf and immature kernel tissues also contained about 1000-fold higher levels of <i>alk</i>-specific small RNAs compared to null controls, indicating that the enhanced aflatoxin resistance in the transgenic maize kernels is due to suppression of <i>A. flavus</i> infection through HIGS of <i>alk</i> gene.
format	article
author	Olanike Omolehin Yenjit Raruang Dongfang Hu Zhu-Qiang Han Qijian Wei Kan Wang Kanniah Rajasekaran Jeffrey W. Cary Zhi-Yuan Chen
author_facet	Olanike Omolehin Yenjit Raruang Dongfang Hu Zhu-Qiang Han Qijian Wei Kan Wang Kanniah Rajasekaran Jeffrey W. Cary Zhi-Yuan Chen
author_sort	Olanike Omolehin
title	Resistance to Aflatoxin Accumulation in Maize Mediated by Host-Induced Silencing of the <i>Aspergillus flavus</i> Alkaline Protease (<i>alk</i>) Gene
title_short	Resistance to Aflatoxin Accumulation in Maize Mediated by Host-Induced Silencing of the <i>Aspergillus flavus</i> Alkaline Protease (<i>alk</i>) Gene
title_full	Resistance to Aflatoxin Accumulation in Maize Mediated by Host-Induced Silencing of the <i>Aspergillus flavus</i> Alkaline Protease (<i>alk</i>) Gene
title_fullStr	Resistance to Aflatoxin Accumulation in Maize Mediated by Host-Induced Silencing of the <i>Aspergillus flavus</i> Alkaline Protease (<i>alk</i>) Gene
title_full_unstemmed	Resistance to Aflatoxin Accumulation in Maize Mediated by Host-Induced Silencing of the <i>Aspergillus flavus</i> Alkaline Protease (<i>alk</i>) Gene
title_sort	resistance to aflatoxin accumulation in maize mediated by host-induced silencing of the <i>aspergillus flavus</i> alkaline protease (<i>alk</i>) gene
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/f3c34a0d00c94124b0e7291fa3af2aed
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Resistance to Aflatoxin Accumulation in Maize Mediated by Host-Induced Silencing of the <i>Aspergillus flavus</i> Alkaline Protease (<i>alk</i>) Gene

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