A Novel Deoxynivalenol-Activated Wheat <i>Arl6ip4</i> Gene Encodes an Antifungal Peptide with Deoxynivalenol Affinity and Protects Plants against <i>Fusarium</i> Pathogens and Mycotoxins
Deoxynivalenol (DON) is one of the most widespread trichothecene mycotoxins in contaminated cereal products. DON plays a vital role in the pathogenesis of <i>Fusarium graminearum</i>, but the molecular mechanisms of DON underlying <i>Fusarium</i>–wheat interactions are not ye...
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2021
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oai:doaj.org-article:9f036fe56a0a46358ede18c3f73c74912021-11-25T18:05:59ZA Novel Deoxynivalenol-Activated Wheat <i>Arl6ip4</i> Gene Encodes an Antifungal Peptide with Deoxynivalenol Affinity and Protects Plants against <i>Fusarium</i> Pathogens and Mycotoxins10.3390/jof71109412309-608Xhttps://doaj.org/article/9f036fe56a0a46358ede18c3f73c74912021-11-01T00:00:00Zhttps://www.mdpi.com/2309-608X/7/11/941https://doaj.org/toc/2309-608XDeoxynivalenol (DON) is one of the most widespread trichothecene mycotoxins in contaminated cereal products. DON plays a vital role in the pathogenesis of <i>Fusarium graminearum</i>, but the molecular mechanisms of DON underlying <i>Fusarium</i>–wheat interactions are not yet well understood. In this study, a novel wheat ADP-ribosylation factor-like protein 6-interacting protein 4 gene, <i>TaArl6ip4,</i> was identified from DON-treated wheat suspension cells by suppression subtractive hybridization (SSH). The qRT-PCR result suggested that <i>TaArl6ip4</i> expression is specifically activated by DON in both the <i>Fusarium</i> intermediate susceptible wheat cultivar Zhengmai9023 and the <i>F</i><i>usarium</i> resistant cultivar Sumai3. The transient expression results of the TaARL6IP4::GFP fusion protein indicate that <i>TaArl6ip4</i> encodes a plasma membrane and nucleus-localized protein. Multiple sequence alignment using microscale thermophoresis showed that TaARL6IP4 comprises a conserved DON binding motif, <sub>67</sub>HXXXG<sub>71</sub>, and exhibits DON affinity with a dissociation constant (<i>K</i><sub>D</sub>) of 91 ± 2.6 µM. Moreover, TaARL6IP4 exhibited antifungal activity with IC<sub>50</sub> values of 22 ± 1.5 µM and 25 ± 2.6 µM against <i>Fusarium graminearum</i> and <i>Alternaria alternata</i>, respectively. Furthermore, TaArl6ip4 interacted with the plasma membrane of <i>Fusarium graminearum</i> spores, resulting in membrane disruption and the leakage of cytoplasmic materials. The heterologous over-expression of <i>TaArl6ip4</i> conferred greater DON tolerance and <i>Fusarium</i> resistance in <i>Arabidopsis.</i> Finally, we describe a novel DON-induced wheat gene, <i>TaArl6ip4</i>, exhibiting antifungal function and DON affinity that may play a key role in <i>Fusarium–</i>wheat interactions.Gang LiuDong-Yun ZuoPeng YangWei-Jie HeZheng YangJing-Bo ZhangAi-Bo WuShu-Yuan YiHe-Ping LiTao HuangYu-Cai LiaoMDPI AGarticledeoxynivalenol<i>Fusarium</i> spp.<i>Triticum aestivum</i>antifungal peptideresistanceplant–pathogen interactionsBiology (General)QH301-705.5ENJournal of Fungi, Vol 7, Iss 941, p 941 (2021) |
| institution |
DOAJ |
| collection |
DOAJ |
| language |
EN |
| topic |
deoxynivalenol <i>Fusarium</i> spp. <i>Triticum aestivum</i> antifungal peptide resistance plant–pathogen interactions Biology (General) QH301-705.5 |
| spellingShingle |
deoxynivalenol <i>Fusarium</i> spp. <i>Triticum aestivum</i> antifungal peptide resistance plant–pathogen interactions Biology (General) QH301-705.5 Gang Liu Dong-Yun Zuo Peng Yang Wei-Jie He Zheng Yang Jing-Bo Zhang Ai-Bo Wu Shu-Yuan Yi He-Ping Li Tao Huang Yu-Cai Liao A Novel Deoxynivalenol-Activated Wheat <i>Arl6ip4</i> Gene Encodes an Antifungal Peptide with Deoxynivalenol Affinity and Protects Plants against <i>Fusarium</i> Pathogens and Mycotoxins |
| description |
Deoxynivalenol (DON) is one of the most widespread trichothecene mycotoxins in contaminated cereal products. DON plays a vital role in the pathogenesis of <i>Fusarium graminearum</i>, but the molecular mechanisms of DON underlying <i>Fusarium</i>–wheat interactions are not yet well understood. In this study, a novel wheat ADP-ribosylation factor-like protein 6-interacting protein 4 gene, <i>TaArl6ip4,</i> was identified from DON-treated wheat suspension cells by suppression subtractive hybridization (SSH). The qRT-PCR result suggested that <i>TaArl6ip4</i> expression is specifically activated by DON in both the <i>Fusarium</i> intermediate susceptible wheat cultivar Zhengmai9023 and the <i>F</i><i>usarium</i> resistant cultivar Sumai3. The transient expression results of the TaARL6IP4::GFP fusion protein indicate that <i>TaArl6ip4</i> encodes a plasma membrane and nucleus-localized protein. Multiple sequence alignment using microscale thermophoresis showed that TaARL6IP4 comprises a conserved DON binding motif, <sub>67</sub>HXXXG<sub>71</sub>, and exhibits DON affinity with a dissociation constant (<i>K</i><sub>D</sub>) of 91 ± 2.6 µM. Moreover, TaARL6IP4 exhibited antifungal activity with IC<sub>50</sub> values of 22 ± 1.5 µM and 25 ± 2.6 µM against <i>Fusarium graminearum</i> and <i>Alternaria alternata</i>, respectively. Furthermore, TaArl6ip4 interacted with the plasma membrane of <i>Fusarium graminearum</i> spores, resulting in membrane disruption and the leakage of cytoplasmic materials. The heterologous over-expression of <i>TaArl6ip4</i> conferred greater DON tolerance and <i>Fusarium</i> resistance in <i>Arabidopsis.</i> Finally, we describe a novel DON-induced wheat gene, <i>TaArl6ip4</i>, exhibiting antifungal function and DON affinity that may play a key role in <i>Fusarium–</i>wheat interactions. |
| format |
article |
| author |
Gang Liu Dong-Yun Zuo Peng Yang Wei-Jie He Zheng Yang Jing-Bo Zhang Ai-Bo Wu Shu-Yuan Yi He-Ping Li Tao Huang Yu-Cai Liao |
| author_facet |
Gang Liu Dong-Yun Zuo Peng Yang Wei-Jie He Zheng Yang Jing-Bo Zhang Ai-Bo Wu Shu-Yuan Yi He-Ping Li Tao Huang Yu-Cai Liao |
| author_sort |
Gang Liu |
| title |
A Novel Deoxynivalenol-Activated Wheat <i>Arl6ip4</i> Gene Encodes an Antifungal Peptide with Deoxynivalenol Affinity and Protects Plants against <i>Fusarium</i> Pathogens and Mycotoxins |
| title_short |
A Novel Deoxynivalenol-Activated Wheat <i>Arl6ip4</i> Gene Encodes an Antifungal Peptide with Deoxynivalenol Affinity and Protects Plants against <i>Fusarium</i> Pathogens and Mycotoxins |
| title_full |
A Novel Deoxynivalenol-Activated Wheat <i>Arl6ip4</i> Gene Encodes an Antifungal Peptide with Deoxynivalenol Affinity and Protects Plants against <i>Fusarium</i> Pathogens and Mycotoxins |
| title_fullStr |
A Novel Deoxynivalenol-Activated Wheat <i>Arl6ip4</i> Gene Encodes an Antifungal Peptide with Deoxynivalenol Affinity and Protects Plants against <i>Fusarium</i> Pathogens and Mycotoxins |
| title_full_unstemmed |
A Novel Deoxynivalenol-Activated Wheat <i>Arl6ip4</i> Gene Encodes an Antifungal Peptide with Deoxynivalenol Affinity and Protects Plants against <i>Fusarium</i> Pathogens and Mycotoxins |
| title_sort |
novel deoxynivalenol-activated wheat <i>arl6ip4</i> gene encodes an antifungal peptide with deoxynivalenol affinity and protects plants against <i>fusarium</i> pathogens and mycotoxins |
| publisher |
MDPI AG |
| publishDate |
2021 |
| url |
https://doaj.org/article/9f036fe56a0a46358ede18c3f73c7491 |
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