Modulation of flavonoid biosynthetic pathway genes and anthocyanins due to virus infection in grapevine (Vitis viniferaL.) leaves
Background: Symptoms of grapevine leafroll disease (GLRD) in red-fruited wine grape (Vitis vinifera L.) cultivars consist of green veins and red and reddish-purple discoloration of inter-veinal areas of leaves. The reddish-purple color of symptomatic leaves may be due to the accumulation of anthocya...
| Autores principales: | , , , |
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| Formato: | Artículo |
| Lenguaje: | Inglés |
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BMC
2020
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| Acceso en línea: | http://hdl.handle.net/20.500.12123/7541 https://bmcplantbiol.biomedcentral.com/articles/10.1186/1471-2229-10-187 https://doi.org/10.1186/1471-2229-10-187 |
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| author | Gutha, Linga R. Casassa, Luis Federico Harbertson, James F. Naidu, Rayapati A. |
| author_browse | Casassa, Luis Federico Gutha, Linga R. Harbertson, James F. Naidu, Rayapati A. |
| author_facet | Gutha, Linga R. Casassa, Luis Federico Harbertson, James F. Naidu, Rayapati A. |
| author_sort | Gutha, Linga R. |
| collection | INTA Digital |
| description | Background: Symptoms of grapevine leafroll disease (GLRD) in red-fruited wine grape (Vitis vinifera L.) cultivars consist of green veins and red and reddish-purple discoloration of inter-veinal areas of leaves. The reddish-purple color of symptomatic leaves may be due to the accumulation of anthocyanins and could reflect an up-regulation of genes involved in their biosynthesis.
Results: We examined six putative constitutively expressed genes, Ubiquitin, Actin, GAPDH, EF1-a, SAND and NAD5, for their potential as references for normalization of gene expression in reverse transcription-quantitative real-time polymerase chain reaction (RT-qPCR). Using the geNorm program, a combination of two genes (Actin and NAD5) was identified as the stable set of reference genes for normalization of gene expression data obtained from grapevine leaves. By using gene-specific RT-qPCR in combination with a reliable normalization factor, we compared relative expression of the flavonoid biosynthetic pathway genes between leaves infected with Grapevine leafroll-associated virus 3 (GLRaV-3) and exhibiting GLRD symptoms and virus-free green leaves obtained from a red-fruited wine grape cultivar (cv. Merlot). The expression levels of these different genes ranged from two- to fifty-fold increase in virus-infected leaves. Among them, CHS3, F3'5'H, F3H1, LDOX, LAR1 and MybA1 showed greater than 10-fold increase suggesting that they were expressed at significantly higher levels in virus-infected symptomatic leaves. HPLC profiling of anthocyanins extracted from leaves indicated the presence of cyanidin-3-glucoside and malvidin-3-glucoside only in virus-infected symptomatic leaves. The results also showed 24% higher levels of flavonols in virus-infected symptomatic leaves than in virus-free green leaves, with quercetin followed by myricetin being the predominant compounds. Proanthocyanidins, estimated as total tannins by protein precipitation method, were 36% higher in virus-infected symptomatic leaves when compared to virus-free green leaves.
Conclusions: The results, the first example to our knowledge, showed that modulation of the flavonoid biosynthetic pathway occurred in GLRaV-3-infected leaves of a red-fruited wine grape cultivar (cv. Merlot) leading to de novo synthesis of two classes of anthocyanins. These anthocyanins have contributed to the expression of reddish-purple color of virus-infected grapevine leaves exhibiting GLRD symptoms. |
| format | Artículo |
| id | INTA7541 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Inglés |
| publishDate | 2020 |
| publishDateRange | 2020 |
| publishDateSort | 2020 |
| publisher | BMC |
| publisherStr | BMC |
| record_format | dspace |
| spelling | INTA75412020-07-13T12:59:00Z Modulation of flavonoid biosynthetic pathway genes and anthocyanins due to virus infection in grapevine (Vitis viniferaL.) leaves Gutha, Linga R. Casassa, Luis Federico Harbertson, James F. Naidu, Rayapati A. Vid Virus de las Plantas Flavonoides Antocianinas Genética Grapevines Plant Viruses Flavonoids Anthocyanins Genetics Background: Symptoms of grapevine leafroll disease (GLRD) in red-fruited wine grape (Vitis vinifera L.) cultivars consist of green veins and red and reddish-purple discoloration of inter-veinal areas of leaves. The reddish-purple color of symptomatic leaves may be due to the accumulation of anthocyanins and could reflect an up-regulation of genes involved in their biosynthesis. Results: We examined six putative constitutively expressed genes, Ubiquitin, Actin, GAPDH, EF1-a, SAND and NAD5, for their potential as references for normalization of gene expression in reverse transcription-quantitative real-time polymerase chain reaction (RT-qPCR). Using the geNorm program, a combination of two genes (Actin and NAD5) was identified as the stable set of reference genes for normalization of gene expression data obtained from grapevine leaves. By using gene-specific RT-qPCR in combination with a reliable normalization factor, we compared relative expression of the flavonoid biosynthetic pathway genes between leaves infected with Grapevine leafroll-associated virus 3 (GLRaV-3) and exhibiting GLRD symptoms and virus-free green leaves obtained from a red-fruited wine grape cultivar (cv. Merlot). The expression levels of these different genes ranged from two- to fifty-fold increase in virus-infected leaves. Among them, CHS3, F3'5'H, F3H1, LDOX, LAR1 and MybA1 showed greater than 10-fold increase suggesting that they were expressed at significantly higher levels in virus-infected symptomatic leaves. HPLC profiling of anthocyanins extracted from leaves indicated the presence of cyanidin-3-glucoside and malvidin-3-glucoside only in virus-infected symptomatic leaves. The results also showed 24% higher levels of flavonols in virus-infected symptomatic leaves than in virus-free green leaves, with quercetin followed by myricetin being the predominant compounds. Proanthocyanidins, estimated as total tannins by protein precipitation method, were 36% higher in virus-infected symptomatic leaves when compared to virus-free green leaves. Conclusions: The results, the first example to our knowledge, showed that modulation of the flavonoid biosynthetic pathway occurred in GLRaV-3-infected leaves of a red-fruited wine grape cultivar (cv. Merlot) leading to de novo synthesis of two classes of anthocyanins. These anthocyanins have contributed to the expression of reddish-purple color of virus-infected grapevine leaves exhibiting GLRD symptoms. EEA Mendoza Fil: Gutha, Linga R. Washington State University. Irrigated Agriculture Research and Extension Center. Department of Plant Pathology; Estados Unidos Fil: Casassa, Luis Federico. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Mendoza. Centro de Estudios de Enología; Argentina. Washington State University. Irrigated Agriculture Research and Extension Center. School of Food Science; Estados Unidos Fil: Harbertson, James F. Washington State University. Irrigated Agriculture Research and Extension Center. School of Food Science; Estados Unidos Fil: Naidu, Rayapati A. Washington State University. Irrigated Agriculture Research and Extension Center. Department of Plant Pathology; Estados Unidos 2020-07-13T12:50:16Z 2020-07-13T12:50:16Z 2010-08 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion http://hdl.handle.net/20.500.12123/7541 https://bmcplantbiol.biomedcentral.com/articles/10.1186/1471-2229-10-187 1471-2229 https://doi.org/10.1186/1471-2229-10-187 eng info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by-nc-sa/4.0/ Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0) application/pdf BMC BMC Plant Biology 10 : article number: 187 (2010) |
| spellingShingle | Vid Virus de las Plantas Flavonoides Antocianinas Genética Grapevines Plant Viruses Flavonoids Anthocyanins Genetics Gutha, Linga R. Casassa, Luis Federico Harbertson, James F. Naidu, Rayapati A. Modulation of flavonoid biosynthetic pathway genes and anthocyanins due to virus infection in grapevine (Vitis viniferaL.) leaves |
| title | Modulation of flavonoid biosynthetic pathway genes and anthocyanins due to virus infection in grapevine (Vitis viniferaL.) leaves |
| title_full | Modulation of flavonoid biosynthetic pathway genes and anthocyanins due to virus infection in grapevine (Vitis viniferaL.) leaves |
| title_fullStr | Modulation of flavonoid biosynthetic pathway genes and anthocyanins due to virus infection in grapevine (Vitis viniferaL.) leaves |
| title_full_unstemmed | Modulation of flavonoid biosynthetic pathway genes and anthocyanins due to virus infection in grapevine (Vitis viniferaL.) leaves |
| title_short | Modulation of flavonoid biosynthetic pathway genes and anthocyanins due to virus infection in grapevine (Vitis viniferaL.) leaves |
| title_sort | modulation of flavonoid biosynthetic pathway genes and anthocyanins due to virus infection in grapevine vitis viniferal leaves |
| topic | Vid Virus de las Plantas Flavonoides Antocianinas Genética Grapevines Plant Viruses Flavonoids Anthocyanins Genetics |
| url | http://hdl.handle.net/20.500.12123/7541 https://bmcplantbiol.biomedcentral.com/articles/10.1186/1471-2229-10-187 https://doi.org/10.1186/1471-2229-10-187 |
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