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β-1,3-glucanase class III promotes spread of PVY(NTN) and improves in planta protein production.

Dobnik D, Baebler S, Kogovšek P, Pompe-Novak M, Stebih D, Panter G, Janež N, Morisset D, Zel J, Gruden K - Plant Biotechnol Rep (2013)

Bottom Line: Differences in viral spread were observed between transgenic lines overexpressing Glu-III and non-transgenic lines, with stronger and faster viral spread in transgenic Désirée, and some multiplication in transgenic Santé.In addition, the ability of Glu-III to improve in planta protein production after agroinfiltration was tested.The results have shown that Glu-III overexpression enables faster spreading of vectors between cells and better protein production, which could be beneficial in improving in planta protein production system using viral vectors.

View Article: PubMed Central - PubMed

Affiliation: Department of Biotechnology and Systems Biology, National Institute of Biology, Večna pot 111, 1000 Ljubljana, Slovenia.

ABSTRACT
Glucanases are enzymes regulating the size exclusion limit and permeability of plasmodesmata and play a role in biotic stress. In plant genomes, they are encoded as relatively large gene families divided into four classes. Most studies of plant virus interactions have focused on glucanases from classes I and II. In our study, we have evaluated the role of the β-1,3-glucanase class III (Glu-III) gene in the potato-potato virus Y(NTN) (PVY(NTN)) interaction and implemented the findings to plant biotechnology application. Potato cultivars Désirée and Santé, which are tolerant and extremely resistant to PVY(NTN), respectively, were stably transformed with Agrobacterium tumefaciens harbouring constructs for Glu-III overexpression. Localization of Glu-III protein in patches within the cell wall was determined by tagging the Glu-III protein with green fluorescent protein. Transgenic and non-transgenic plants were challenged with PVY(NTN) and its multiplication and spreading was followed. Differences in viral spread were observed between transgenic lines overexpressing Glu-III and non-transgenic lines, with stronger and faster viral spread in transgenic Désirée, and some multiplication in transgenic Santé. In addition, the ability of Glu-III to improve in planta protein production after agroinfiltration was tested. The results have shown that Glu-III overexpression enables faster spreading of vectors between cells and better protein production, which could be beneficial in improving in planta protein production system using viral vectors.

No MeSH data available.


Related in: MedlinePlus

Effect of Glu-III overexpression on multiplication and spread of PVYNTN in cv. Désirée. Results of two separate experiments are shown. PVYNTN content was followed in upper non-inoculated leaves to monitor long-distance movement of the virus. Numbers 1–6 denote individual plants. In plants where no column is visible, PVYNTN was not detected. WD non-transgenic potato cv. Désirée, TD transgenic potato cv. Désirée
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Fig1: Effect of Glu-III overexpression on multiplication and spread of PVYNTN in cv. Désirée. Results of two separate experiments are shown. PVYNTN content was followed in upper non-inoculated leaves to monitor long-distance movement of the virus. Numbers 1–6 denote individual plants. In plants where no column is visible, PVYNTN was not detected. WD non-transgenic potato cv. Désirée, TD transgenic potato cv. Désirée

Mentions: Virus multiplication and its systemic spreading in the tolerant cultivar Désirée was studied in transgenic Désirée plants overexpressing Glu-III and compared with that in non-transgenic counterparts. In the first experiment, multiplication of the virus was followed in the lower, inoculated leaves, with eventual systemic spread of the virus after 1, 4. and 7 dpi. In the second experiment, spread of the virus to non-inoculated leaves (systemic infection) was monitored at later time points after infection (7, 10. and 13 dpi). No statistically significant difference in virus multiplication was observed between transgenic and non-transgenic Désirée in the inoculated leaves, nor any significant increase in the amount of viral RNA over time (Fig. S3). Viral RNA was not detected in upper, non-inoculated leaves at 1 and 4 dpi. At 7 dpi, viral RNA was detected in 2 of 6 plants in both Désirée genotypes (Fig. 1). No statistically significant difference was observed in the amounts of viral RNA in transgenic and non-transgenic lines of Désirée in non-inoculated leaves.Fig. 1


β-1,3-glucanase class III promotes spread of PVY(NTN) and improves in planta protein production.

Dobnik D, Baebler S, Kogovšek P, Pompe-Novak M, Stebih D, Panter G, Janež N, Morisset D, Zel J, Gruden K - Plant Biotechnol Rep (2013)

Effect of Glu-III overexpression on multiplication and spread of PVYNTN in cv. Désirée. Results of two separate experiments are shown. PVYNTN content was followed in upper non-inoculated leaves to monitor long-distance movement of the virus. Numbers 1–6 denote individual plants. In plants where no column is visible, PVYNTN was not detected. WD non-transgenic potato cv. Désirée, TD transgenic potato cv. Désirée
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3824212&req=5

Fig1: Effect of Glu-III overexpression on multiplication and spread of PVYNTN in cv. Désirée. Results of two separate experiments are shown. PVYNTN content was followed in upper non-inoculated leaves to monitor long-distance movement of the virus. Numbers 1–6 denote individual plants. In plants where no column is visible, PVYNTN was not detected. WD non-transgenic potato cv. Désirée, TD transgenic potato cv. Désirée
Mentions: Virus multiplication and its systemic spreading in the tolerant cultivar Désirée was studied in transgenic Désirée plants overexpressing Glu-III and compared with that in non-transgenic counterparts. In the first experiment, multiplication of the virus was followed in the lower, inoculated leaves, with eventual systemic spread of the virus after 1, 4. and 7 dpi. In the second experiment, spread of the virus to non-inoculated leaves (systemic infection) was monitored at later time points after infection (7, 10. and 13 dpi). No statistically significant difference in virus multiplication was observed between transgenic and non-transgenic Désirée in the inoculated leaves, nor any significant increase in the amount of viral RNA over time (Fig. S3). Viral RNA was not detected in upper, non-inoculated leaves at 1 and 4 dpi. At 7 dpi, viral RNA was detected in 2 of 6 plants in both Désirée genotypes (Fig. 1). No statistically significant difference was observed in the amounts of viral RNA in transgenic and non-transgenic lines of Désirée in non-inoculated leaves.Fig. 1

Bottom Line: Differences in viral spread were observed between transgenic lines overexpressing Glu-III and non-transgenic lines, with stronger and faster viral spread in transgenic Désirée, and some multiplication in transgenic Santé.In addition, the ability of Glu-III to improve in planta protein production after agroinfiltration was tested.The results have shown that Glu-III overexpression enables faster spreading of vectors between cells and better protein production, which could be beneficial in improving in planta protein production system using viral vectors.

View Article: PubMed Central - PubMed

Affiliation: Department of Biotechnology and Systems Biology, National Institute of Biology, Večna pot 111, 1000 Ljubljana, Slovenia.

ABSTRACT
Glucanases are enzymes regulating the size exclusion limit and permeability of plasmodesmata and play a role in biotic stress. In plant genomes, they are encoded as relatively large gene families divided into four classes. Most studies of plant virus interactions have focused on glucanases from classes I and II. In our study, we have evaluated the role of the β-1,3-glucanase class III (Glu-III) gene in the potato-potato virus Y(NTN) (PVY(NTN)) interaction and implemented the findings to plant biotechnology application. Potato cultivars Désirée and Santé, which are tolerant and extremely resistant to PVY(NTN), respectively, were stably transformed with Agrobacterium tumefaciens harbouring constructs for Glu-III overexpression. Localization of Glu-III protein in patches within the cell wall was determined by tagging the Glu-III protein with green fluorescent protein. Transgenic and non-transgenic plants were challenged with PVY(NTN) and its multiplication and spreading was followed. Differences in viral spread were observed between transgenic lines overexpressing Glu-III and non-transgenic lines, with stronger and faster viral spread in transgenic Désirée, and some multiplication in transgenic Santé. In addition, the ability of Glu-III to improve in planta protein production after agroinfiltration was tested. The results have shown that Glu-III overexpression enables faster spreading of vectors between cells and better protein production, which could be beneficial in improving in planta protein production system using viral vectors.

No MeSH data available.


Related in: MedlinePlus