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Effect of Potato virus Y on the NADP-malic enzyme from Nicotiana tabacum L.: mRNA, expressed protein and activity.

Doubnerová V, Müller K, Cerovská N, Synková H, Spoustová P, Ryslavá H - Int J Mol Sci (2009)

Bottom Line: Petit Havana, SR1) was tested at the transcriptional, translational and activity level.The effect of viral infection induced by Potato virus Y, NTN strain, causing more severe symptoms, was compared with the effect induced by milder strain PVY(O).The relevance of NADP-malic enzyme in plants under stress conditions was discussed.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry, Charles University, Prague, Czech Republic. doubnerova@volny.cz <doubnerova@volny.cz>

ABSTRACT
The effect of biotic stress induced by viral infection (Potato virus Y, strain NTN and O) on NADP-malic enzyme (EC 1.1.1.40) in tobacco plants (Nicotiana tabacum L., cv. Petit Havana, SR1) was tested at the transcriptional, translational and activity level. The increase of enzyme activity in infected leaves was correlated with the increased amount of expressed protein and with mRNA of cytosolic NADP-ME isoform. Transcription of the chloroplastic enzyme was not influenced by viral infection. The increase of the enzyme activity was also detected in stems and roots of infected plants. The effect of viral infection induced by Potato virus Y, NTN strain, causing more severe symptoms, was compared with the effect induced by milder strain PVY(O). The observed increase in NADP-malic enzyme activity in all parts of the studied plants was higher in the case of PVY(NTN) strain than in the case of strain PVY(O). The relevance of NADP-malic enzyme in plants under stress conditions was discussed.

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Related in: MedlinePlus

Detection of NADP-ME in 10% polyacrylamide gel after non-denaturating electrophoresis. Control (marked as C), PVYO and PVYNTN infected leaves, stems and roots of Nicotiana tabacum L. plants were analysed. Samples were collected at the maximal symptoms occurrence (11th day after PVYO inoculation and 13th day after PVYNTN inoculation).
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f4-ijms-10-03583: Detection of NADP-ME in 10% polyacrylamide gel after non-denaturating electrophoresis. Control (marked as C), PVYO and PVYNTN infected leaves, stems and roots of Nicotiana tabacum L. plants were analysed. Samples were collected at the maximal symptoms occurrence (11th day after PVYO inoculation and 13th day after PVYNTN inoculation).

Mentions: NADP-ME activity was also evaluated after electrophoretic separation in polyacrylamide gel under native conditions. Higher NADP-ME activity in infected plants corresponded to more intensive bands noticeable after its detection (Figure 4). Active NADP-ME zones in gel regarding to control, PVYO and PVYNTN infected leaves; stems and roots are shown in Figure 4. One band was visible in all parts (leaves, stems and roots) of Nicotiana tabacum L. (Figure 4). The estimation of relative molecular mass after native electrophoresis was performed according to Ferguson’s method (using a set of various polyacrylamide gel concentrations) and was approximately 264,000. This molecular weight would correspond to tetramer and to the cytosolic NADP-ME. Particularly isoforms could not be distinguished, probably due to that they have similar molecular weights or due to low activity of chloroplastic isoform (data not shown). No band of NADP-ME activity was observed in control gel incubations without substrate, coenzyme and cofactor, respectively (results not shown). Different results were found in Nicotiana benthamiana plants, where PVYNTN did not cause a significant increase of the NADP-ME activity in leaves, but only a slight increase (1.8-fold higher) of NADP-ME activity in roots [22]. Nicotiana benthamiana and Nicotiana tabacum L. plants also differed in NADP-ME isoform content. Generally, we can conclude, that enhanced activity of NADP-ME was probably not a general response to potyviral infection in all host plants, seeing that the presence of isoforms and their functions might be also important.


Effect of Potato virus Y on the NADP-malic enzyme from Nicotiana tabacum L.: mRNA, expressed protein and activity.

Doubnerová V, Müller K, Cerovská N, Synková H, Spoustová P, Ryslavá H - Int J Mol Sci (2009)

Detection of NADP-ME in 10% polyacrylamide gel after non-denaturating electrophoresis. Control (marked as C), PVYO and PVYNTN infected leaves, stems and roots of Nicotiana tabacum L. plants were analysed. Samples were collected at the maximal symptoms occurrence (11th day after PVYO inoculation and 13th day after PVYNTN inoculation).
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC2812832&req=5

f4-ijms-10-03583: Detection of NADP-ME in 10% polyacrylamide gel after non-denaturating electrophoresis. Control (marked as C), PVYO and PVYNTN infected leaves, stems and roots of Nicotiana tabacum L. plants were analysed. Samples were collected at the maximal symptoms occurrence (11th day after PVYO inoculation and 13th day after PVYNTN inoculation).
Mentions: NADP-ME activity was also evaluated after electrophoretic separation in polyacrylamide gel under native conditions. Higher NADP-ME activity in infected plants corresponded to more intensive bands noticeable after its detection (Figure 4). Active NADP-ME zones in gel regarding to control, PVYO and PVYNTN infected leaves; stems and roots are shown in Figure 4. One band was visible in all parts (leaves, stems and roots) of Nicotiana tabacum L. (Figure 4). The estimation of relative molecular mass after native electrophoresis was performed according to Ferguson’s method (using a set of various polyacrylamide gel concentrations) and was approximately 264,000. This molecular weight would correspond to tetramer and to the cytosolic NADP-ME. Particularly isoforms could not be distinguished, probably due to that they have similar molecular weights or due to low activity of chloroplastic isoform (data not shown). No band of NADP-ME activity was observed in control gel incubations without substrate, coenzyme and cofactor, respectively (results not shown). Different results were found in Nicotiana benthamiana plants, where PVYNTN did not cause a significant increase of the NADP-ME activity in leaves, but only a slight increase (1.8-fold higher) of NADP-ME activity in roots [22]. Nicotiana benthamiana and Nicotiana tabacum L. plants also differed in NADP-ME isoform content. Generally, we can conclude, that enhanced activity of NADP-ME was probably not a general response to potyviral infection in all host plants, seeing that the presence of isoforms and their functions might be also important.

Bottom Line: Petit Havana, SR1) was tested at the transcriptional, translational and activity level.The effect of viral infection induced by Potato virus Y, NTN strain, causing more severe symptoms, was compared with the effect induced by milder strain PVY(O).The relevance of NADP-malic enzyme in plants under stress conditions was discussed.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry, Charles University, Prague, Czech Republic. doubnerova@volny.cz <doubnerova@volny.cz>

ABSTRACT
The effect of biotic stress induced by viral infection (Potato virus Y, strain NTN and O) on NADP-malic enzyme (EC 1.1.1.40) in tobacco plants (Nicotiana tabacum L., cv. Petit Havana, SR1) was tested at the transcriptional, translational and activity level. The increase of enzyme activity in infected leaves was correlated with the increased amount of expressed protein and with mRNA of cytosolic NADP-ME isoform. Transcription of the chloroplastic enzyme was not influenced by viral infection. The increase of the enzyme activity was also detected in stems and roots of infected plants. The effect of viral infection induced by Potato virus Y, NTN strain, causing more severe symptoms, was compared with the effect induced by milder strain PVY(O). The observed increase in NADP-malic enzyme activity in all parts of the studied plants was higher in the case of PVY(NTN) strain than in the case of strain PVY(O). The relevance of NADP-malic enzyme in plants under stress conditions was discussed.

Show MeSH
Related in: MedlinePlus