A pathogenesis related protein, VpPR-10.1, from Vitis pseudoreticulata: an insight of its mode of antifungal activity.
Previously, VpPR-10.1 was isolated and characterized from a cDNA library of a fungus-resistant accession of Chinese wild grape (Vitis pseudoreticulata). We found that expression of VpPR-10.1 is affected by the fungal pathogen Erysiphe necator. To investigate the biochemical basis of the nuclease act...
Saved in:
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2014-01-01
|
| Series: | PLoS ONE |
| Online Access: | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0095102&type=printable |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1850191675139293184 |
|---|---|
| author | Teng-Fei Xu Xiao-Chen Zhao Yun-Tong Jiao Jin-Yu Wei Lan Wang Yan Xu |
| author_facet | Teng-Fei Xu Xiao-Chen Zhao Yun-Tong Jiao Jin-Yu Wei Lan Wang Yan Xu |
| author_sort | Teng-Fei Xu |
| collection | DOAJ |
| description | Previously, VpPR-10.1 was isolated and characterized from a cDNA library of a fungus-resistant accession of Chinese wild grape (Vitis pseudoreticulata). We found that expression of VpPR-10.1 is affected by the fungal pathogen Erysiphe necator. To investigate the biochemical basis of the nuclease activity of VpPR-10.1 and its role in antifungal resistance, we generated recombinant VpPR-10.1 as well as site-directed mutations targeting three conserved amino acid residues among plant PR-10 s: Lys55, Glu149, and Tyr151. We showed that wild-type recombinant VpPR-10.1 exhibits both RNase and DNase activities. Mutant VpPR10.1-Y151H essentially retained all these activities. In contrast, VpPR10.1-K55N, where Lys55 in the P-loop region is mutated to Asn, and VpPR10.1-E149G, where Glu149 is mutated to Gly, lost their nuclease activity, indicating that both residues play a critical role in catalyzing RNA and DNA degradation. Furthermore, VpPR10.1 and VpPR10.1-Y151H inhibited the growth of the cultured fungal pathogen Alternaria alternate. Through transient expression in grapevine, we also demonstrated that VpPR10.1-K55N and VpPR10.1-E149G compromised resistance to E. necator. Finally, we further found that VpPR-10.1 can lead to programmed cell death and DNA degradation when incubated with tobacco BY-2 suspension cells. We show here that Lys55 and Glu149, but not Tyr151, are required for the RNase, DNase and antifungal activities of VpPR-10.1. The strong correlation between the level of VpPR-10.1 nuclease activity and its antifungal property indicates that the former is the biochemical basis for the latter. Taken together, our experiments revealed that VpPR-10.1 is critical in mediating fungal resistance in grape, potentially playing a dual role by degrading pathogen RNA and inducing programmed death of host cells. |
| format | Article |
| id | doaj-art-3edc81df056045a8a430ad0523c954aa |
| institution | OA Journals |
| issn | 1932-6203 |
| language | English |
| publishDate | 2014-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj-art-3edc81df056045a8a430ad0523c954aa2025-08-20T02:14:50ZengPublic Library of Science (PLoS)PLoS ONE1932-62032014-01-0194e9510210.1371/journal.pone.0095102A pathogenesis related protein, VpPR-10.1, from Vitis pseudoreticulata: an insight of its mode of antifungal activity.Teng-Fei XuXiao-Chen ZhaoYun-Tong JiaoJin-Yu WeiLan WangYan XuPreviously, VpPR-10.1 was isolated and characterized from a cDNA library of a fungus-resistant accession of Chinese wild grape (Vitis pseudoreticulata). We found that expression of VpPR-10.1 is affected by the fungal pathogen Erysiphe necator. To investigate the biochemical basis of the nuclease activity of VpPR-10.1 and its role in antifungal resistance, we generated recombinant VpPR-10.1 as well as site-directed mutations targeting three conserved amino acid residues among plant PR-10 s: Lys55, Glu149, and Tyr151. We showed that wild-type recombinant VpPR-10.1 exhibits both RNase and DNase activities. Mutant VpPR10.1-Y151H essentially retained all these activities. In contrast, VpPR10.1-K55N, where Lys55 in the P-loop region is mutated to Asn, and VpPR10.1-E149G, where Glu149 is mutated to Gly, lost their nuclease activity, indicating that both residues play a critical role in catalyzing RNA and DNA degradation. Furthermore, VpPR10.1 and VpPR10.1-Y151H inhibited the growth of the cultured fungal pathogen Alternaria alternate. Through transient expression in grapevine, we also demonstrated that VpPR10.1-K55N and VpPR10.1-E149G compromised resistance to E. necator. Finally, we further found that VpPR-10.1 can lead to programmed cell death and DNA degradation when incubated with tobacco BY-2 suspension cells. We show here that Lys55 and Glu149, but not Tyr151, are required for the RNase, DNase and antifungal activities of VpPR-10.1. The strong correlation between the level of VpPR-10.1 nuclease activity and its antifungal property indicates that the former is the biochemical basis for the latter. Taken together, our experiments revealed that VpPR-10.1 is critical in mediating fungal resistance in grape, potentially playing a dual role by degrading pathogen RNA and inducing programmed death of host cells.https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0095102&type=printable |
| spellingShingle | Teng-Fei Xu Xiao-Chen Zhao Yun-Tong Jiao Jin-Yu Wei Lan Wang Yan Xu A pathogenesis related protein, VpPR-10.1, from Vitis pseudoreticulata: an insight of its mode of antifungal activity. PLoS ONE |
| title | A pathogenesis related protein, VpPR-10.1, from Vitis pseudoreticulata: an insight of its mode of antifungal activity. |
| title_full | A pathogenesis related protein, VpPR-10.1, from Vitis pseudoreticulata: an insight of its mode of antifungal activity. |
| title_fullStr | A pathogenesis related protein, VpPR-10.1, from Vitis pseudoreticulata: an insight of its mode of antifungal activity. |
| title_full_unstemmed | A pathogenesis related protein, VpPR-10.1, from Vitis pseudoreticulata: an insight of its mode of antifungal activity. |
| title_short | A pathogenesis related protein, VpPR-10.1, from Vitis pseudoreticulata: an insight of its mode of antifungal activity. |
| title_sort | pathogenesis related protein vppr 10 1 from vitis pseudoreticulata an insight of its mode of antifungal activity |
| url | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0095102&type=printable |
| work_keys_str_mv | AT tengfeixu apathogenesisrelatedproteinvppr101fromvitispseudoreticulataaninsightofitsmodeofantifungalactivity AT xiaochenzhao apathogenesisrelatedproteinvppr101fromvitispseudoreticulataaninsightofitsmodeofantifungalactivity AT yuntongjiao apathogenesisrelatedproteinvppr101fromvitispseudoreticulataaninsightofitsmodeofantifungalactivity AT jinyuwei apathogenesisrelatedproteinvppr101fromvitispseudoreticulataaninsightofitsmodeofantifungalactivity AT lanwang apathogenesisrelatedproteinvppr101fromvitispseudoreticulataaninsightofitsmodeofantifungalactivity AT yanxu apathogenesisrelatedproteinvppr101fromvitispseudoreticulataaninsightofitsmodeofantifungalactivity AT tengfeixu pathogenesisrelatedproteinvppr101fromvitispseudoreticulataaninsightofitsmodeofantifungalactivity AT xiaochenzhao pathogenesisrelatedproteinvppr101fromvitispseudoreticulataaninsightofitsmodeofantifungalactivity AT yuntongjiao pathogenesisrelatedproteinvppr101fromvitispseudoreticulataaninsightofitsmodeofantifungalactivity AT jinyuwei pathogenesisrelatedproteinvppr101fromvitispseudoreticulataaninsightofitsmodeofantifungalactivity AT lanwang pathogenesisrelatedproteinvppr101fromvitispseudoreticulataaninsightofitsmodeofantifungalactivity AT yanxu pathogenesisrelatedproteinvppr101fromvitispseudoreticulataaninsightofitsmodeofantifungalactivity |