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Characterization and expression profile of CaNAC2 pepper gene.

Guo WL, Wang SB, Chen RG, Chen BH, Du XH, Yin YX, Gong ZH, Zhang YY - Front Plant Sci (2015)

Bottom Line: Our results indicated that a plant-specific and conserved NAC domain was located in the N-terminus domain of CaNAC2 which was predicted to encode a polypeptide of 410 amino acids.CaNAC2 was expressed mainly in seed and root.These results indicated that this novel CaNAC2 gene might be involved in pepper response to abiotic stress tolerance.

View Article: PubMed Central - PubMed

Affiliation: College of Horticulture, Northwest A&F University Yangling, China ; School of Horticulture Landscape Architecture, Henan Institute of Science and Technology Xinxiang, China.

ABSTRACT
The plant-specific NAC (NAM, ATAF, and CUC) transcription factors have diverse role in development and stress regulation. A new transcript encoding NAC protein, homologous to nam-like protein 4 from Petunia was identified from an ABA-regulated subtractive cDNA library of Capsicum annuum seedling. Here, this homolog (named CaNAC2) from C. annuum was characterized and investigated its role in abiotic stress tolerance. Our results indicated that a plant-specific and conserved NAC domain was located in the N-terminus domain of CaNAC2 which was predicted to encode a polypeptide of 410 amino acids. Phylogenetic analysis showed that CaNAC2 belonged to the NAC2 subgroup of the orthologous group 4d. The protein CaNAC2 was subcellularly localized in the nucleus and it had transcriptional activity in yeast cell. CaNAC2 was expressed mainly in seed and root. The transcription expression of CaNAC2 was strongly induced by cold, salt and ABA treatment and inhibited by osmotic stress and SA treatment. Silence of CaNAC2 in virus-induced gene silenced pepper seedlings resulted in the increased susceptibility to cold stress and delayed the salt-induced leaf chlorophyll degradation. These results indicated that this novel CaNAC2 gene might be involved in pepper response to abiotic stress tolerance.

No MeSH data available.


Related in: MedlinePlus

The subcellular localization and transcriptional activation analysis of pepper CaNAC2. (A) The subcellular localization of pepper CaNAC2 in onion epidermal cells. The fused pBI221-GFP-CaNAC2 and pBI221-GFP constructs were introduced into onion epidermal cells by biolistic bombardment. The GFP signals were observed under confocal microscope; (B) Transcriptional activation analysis of CaNAC2 in yeast strain AH109. CaNAC2 represents the fusion protein of the GAL4 DNA-binding domain and CaNAC2; pGBKT7 was used as control. The culture solution of the transformed yeast was streaked on SD/-Leu/-Trp medium and SD/-Ade/-His/-Leu/-Trp medium. The plates were incubated for 3 days.
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Figure 2: The subcellular localization and transcriptional activation analysis of pepper CaNAC2. (A) The subcellular localization of pepper CaNAC2 in onion epidermal cells. The fused pBI221-GFP-CaNAC2 and pBI221-GFP constructs were introduced into onion epidermal cells by biolistic bombardment. The GFP signals were observed under confocal microscope; (B) Transcriptional activation analysis of CaNAC2 in yeast strain AH109. CaNAC2 represents the fusion protein of the GAL4 DNA-binding domain and CaNAC2; pGBKT7 was used as control. The culture solution of the transformed yeast was streaked on SD/-Leu/-Trp medium and SD/-Ade/-His/-Leu/-Trp medium. The plates were incubated for 3 days.

Mentions: CaNAC2 was predicted to be nucleus localized by ProComp v 8.0 program. To confirm this prediction, the subcellular localization of CaNAC2-GFP fusion protein was demonstrated by fusing the entire CaNAC2 coding sequence to GFP and expressed them under the control of the CaMV35S promoter in the onion epidermal cells. Cells expressing GFP alone displayed diffuse cytoplasmic and nuclear staining (Figure 2A) supported by Guo et al. (2015), while the GFP-CaNAC2 full length protein was located only in the nucleus and the 132–158 amino acids containing the putative NLS can mediate the nuclear targeting of the protein.


Characterization and expression profile of CaNAC2 pepper gene.

Guo WL, Wang SB, Chen RG, Chen BH, Du XH, Yin YX, Gong ZH, Zhang YY - Front Plant Sci (2015)

The subcellular localization and transcriptional activation analysis of pepper CaNAC2. (A) The subcellular localization of pepper CaNAC2 in onion epidermal cells. The fused pBI221-GFP-CaNAC2 and pBI221-GFP constructs were introduced into onion epidermal cells by biolistic bombardment. The GFP signals were observed under confocal microscope; (B) Transcriptional activation analysis of CaNAC2 in yeast strain AH109. CaNAC2 represents the fusion protein of the GAL4 DNA-binding domain and CaNAC2; pGBKT7 was used as control. The culture solution of the transformed yeast was streaked on SD/-Leu/-Trp medium and SD/-Ade/-His/-Leu/-Trp medium. The plates were incubated for 3 days.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4585251&req=5

Figure 2: The subcellular localization and transcriptional activation analysis of pepper CaNAC2. (A) The subcellular localization of pepper CaNAC2 in onion epidermal cells. The fused pBI221-GFP-CaNAC2 and pBI221-GFP constructs were introduced into onion epidermal cells by biolistic bombardment. The GFP signals were observed under confocal microscope; (B) Transcriptional activation analysis of CaNAC2 in yeast strain AH109. CaNAC2 represents the fusion protein of the GAL4 DNA-binding domain and CaNAC2; pGBKT7 was used as control. The culture solution of the transformed yeast was streaked on SD/-Leu/-Trp medium and SD/-Ade/-His/-Leu/-Trp medium. The plates were incubated for 3 days.
Mentions: CaNAC2 was predicted to be nucleus localized by ProComp v 8.0 program. To confirm this prediction, the subcellular localization of CaNAC2-GFP fusion protein was demonstrated by fusing the entire CaNAC2 coding sequence to GFP and expressed them under the control of the CaMV35S promoter in the onion epidermal cells. Cells expressing GFP alone displayed diffuse cytoplasmic and nuclear staining (Figure 2A) supported by Guo et al. (2015), while the GFP-CaNAC2 full length protein was located only in the nucleus and the 132–158 amino acids containing the putative NLS can mediate the nuclear targeting of the protein.

Bottom Line: Our results indicated that a plant-specific and conserved NAC domain was located in the N-terminus domain of CaNAC2 which was predicted to encode a polypeptide of 410 amino acids.CaNAC2 was expressed mainly in seed and root.These results indicated that this novel CaNAC2 gene might be involved in pepper response to abiotic stress tolerance.

View Article: PubMed Central - PubMed

Affiliation: College of Horticulture, Northwest A&F University Yangling, China ; School of Horticulture Landscape Architecture, Henan Institute of Science and Technology Xinxiang, China.

ABSTRACT
The plant-specific NAC (NAM, ATAF, and CUC) transcription factors have diverse role in development and stress regulation. A new transcript encoding NAC protein, homologous to nam-like protein 4 from Petunia was identified from an ABA-regulated subtractive cDNA library of Capsicum annuum seedling. Here, this homolog (named CaNAC2) from C. annuum was characterized and investigated its role in abiotic stress tolerance. Our results indicated that a plant-specific and conserved NAC domain was located in the N-terminus domain of CaNAC2 which was predicted to encode a polypeptide of 410 amino acids. Phylogenetic analysis showed that CaNAC2 belonged to the NAC2 subgroup of the orthologous group 4d. The protein CaNAC2 was subcellularly localized in the nucleus and it had transcriptional activity in yeast cell. CaNAC2 was expressed mainly in seed and root. The transcription expression of CaNAC2 was strongly induced by cold, salt and ABA treatment and inhibited by osmotic stress and SA treatment. Silence of CaNAC2 in virus-induced gene silenced pepper seedlings resulted in the increased susceptibility to cold stress and delayed the salt-induced leaf chlorophyll degradation. These results indicated that this novel CaNAC2 gene might be involved in pepper response to abiotic stress tolerance.

No MeSH data available.


Related in: MedlinePlus