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AaERF1 positively regulates the resistance to Botrytis cinerea in Artemisia annua.

Lu X, Jiang W, Zhang L, Zhang F, Zhang F, Shen Q, Wang G, Tang K - PLoS ONE (2013)

Bottom Line: The results of electrophoretic mobility shift assay (EMSA) and yeast one-hybrid experiments showed that AaERF1 was able to bind to the GCC box cis-acting element in vitro and in yeast.The down-regulated expression level of AaERF1 evidently reduced the resistance to B. cinerea in A. annua.The overall results showed that AaERF1 positively regulated the resistance to B. cinerea in A. annua.

View Article: PubMed Central - PubMed

Affiliation: Fudan-SJTU-Nottingham Plant Biotechnology R&D Center, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, People's Republic of China.

ABSTRACT
Plants are sessile organisms, and they can not move away under abiotic or biotic stresses. Thus plants have evolved a set of genes that response to adverse environment to modulate gene expression. In this study, we characterized and functionally studied an ERF transcription factor from Artemisia annua, AaERF1, which plays an important role in biotic stress responses. The AaERF1 promoter had been cloned and GUS staining results of AaERF1 promoter-GUS transgenic A. annua showed that AaERF1 is expressed ubiquitiously in all organs. Several putative cis-acting elements such as W-box, TGA-box and Py-rich element, which are involved in defense responsiveness, are present in the promoter. The expression of AaERF1 can be induced vigorously by methyl jasmonate as well as by ethephon and wounding, implying that AaERF1 may activate some of the defense genes via the jasmonic acid and ethylene signaling pathways of A. annua. The results of electrophoretic mobility shift assay (EMSA) and yeast one-hybrid experiments showed that AaERF1 was able to bind to the GCC box cis-acting element in vitro and in yeast. Ectopic expression of AaERF1 could enhance the expression levels of the defense marker genes PLANT DEFENSIN1.2 (PDF1.2) and BASIC CHITINASE (ChiB), and increase the resistance to Botrytis cinerea in the 35S::AaERF1 transgenic Arabidopsis. The down-regulated expression level of AaERF1 evidently reduced the resistance to B. cinerea in A. annua. The overall results showed that AaERF1 positively regulated the resistance to B. cinerea in A. annua.

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The expression levels of AaERF1, Chi-B and PDF1.2 in 35S::AaERF1 transgenic Arabidopsis analyzed by RT-Q-PCR.Vertical bars represent standard deviation. A. The expression of AaERF1 in the control and transgenic Arabidopsis plants. Values indicate the mean fold relative to sample the AaERF1-5 transgenic plants. B. The expression of Chi-B in the control and transgenic Arabidopsis plants. Values indicate the mean fold relative to sample the pCAMBIA2300+ empty vector transgenic plants C. The expression of PDF1.2 in the control and transgenic Arabidopsis plants. Values indicate the mean fold relative to sample the pCAMBIA2300+ empty vector transgenic plants. Actin is used as a control for normalization. Data are averages ± SE from three independent experiments.
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pone-0057657-g005: The expression levels of AaERF1, Chi-B and PDF1.2 in 35S::AaERF1 transgenic Arabidopsis analyzed by RT-Q-PCR.Vertical bars represent standard deviation. A. The expression of AaERF1 in the control and transgenic Arabidopsis plants. Values indicate the mean fold relative to sample the AaERF1-5 transgenic plants. B. The expression of Chi-B in the control and transgenic Arabidopsis plants. Values indicate the mean fold relative to sample the pCAMBIA2300+ empty vector transgenic plants C. The expression of PDF1.2 in the control and transgenic Arabidopsis plants. Values indicate the mean fold relative to sample the pCAMBIA2300+ empty vector transgenic plants. Actin is used as a control for normalization. Data are averages ± SE from three independent experiments.

Mentions: The transgenic Arabidopsis plants were first confirmed by kanamycin-resistant screening and genomic DNA-based PCR, and then three transgenic lines were chosen for further analysis. The control experiment involving the transfer of empty plasmid p2300+ to Arabidopsis was also conducted. The results showed that the transcript levels of AaERF1 had a significant increase in AaERF1-overexpression lines (Figure 5A). Correspondingly, Chi-B was shown to be elevated between 2.3- and 7.7-fold in AaERF1-overexpression lines (Figure 5B). The transcript levels of PDF1.2 were elevated between 15- and 1269-fold than that of the control (Figure 5C). The statistics analysis showed that the observed differences were statistically significant.


AaERF1 positively regulates the resistance to Botrytis cinerea in Artemisia annua.

Lu X, Jiang W, Zhang L, Zhang F, Zhang F, Shen Q, Wang G, Tang K - PLoS ONE (2013)

The expression levels of AaERF1, Chi-B and PDF1.2 in 35S::AaERF1 transgenic Arabidopsis analyzed by RT-Q-PCR.Vertical bars represent standard deviation. A. The expression of AaERF1 in the control and transgenic Arabidopsis plants. Values indicate the mean fold relative to sample the AaERF1-5 transgenic plants. B. The expression of Chi-B in the control and transgenic Arabidopsis plants. Values indicate the mean fold relative to sample the pCAMBIA2300+ empty vector transgenic plants C. The expression of PDF1.2 in the control and transgenic Arabidopsis plants. Values indicate the mean fold relative to sample the pCAMBIA2300+ empty vector transgenic plants. Actin is used as a control for normalization. Data are averages ± SE from three independent experiments.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3585223&req=5

pone-0057657-g005: The expression levels of AaERF1, Chi-B and PDF1.2 in 35S::AaERF1 transgenic Arabidopsis analyzed by RT-Q-PCR.Vertical bars represent standard deviation. A. The expression of AaERF1 in the control and transgenic Arabidopsis plants. Values indicate the mean fold relative to sample the AaERF1-5 transgenic plants. B. The expression of Chi-B in the control and transgenic Arabidopsis plants. Values indicate the mean fold relative to sample the pCAMBIA2300+ empty vector transgenic plants C. The expression of PDF1.2 in the control and transgenic Arabidopsis plants. Values indicate the mean fold relative to sample the pCAMBIA2300+ empty vector transgenic plants. Actin is used as a control for normalization. Data are averages ± SE from three independent experiments.
Mentions: The transgenic Arabidopsis plants were first confirmed by kanamycin-resistant screening and genomic DNA-based PCR, and then three transgenic lines were chosen for further analysis. The control experiment involving the transfer of empty plasmid p2300+ to Arabidopsis was also conducted. The results showed that the transcript levels of AaERF1 had a significant increase in AaERF1-overexpression lines (Figure 5A). Correspondingly, Chi-B was shown to be elevated between 2.3- and 7.7-fold in AaERF1-overexpression lines (Figure 5B). The transcript levels of PDF1.2 were elevated between 15- and 1269-fold than that of the control (Figure 5C). The statistics analysis showed that the observed differences were statistically significant.

Bottom Line: The results of electrophoretic mobility shift assay (EMSA) and yeast one-hybrid experiments showed that AaERF1 was able to bind to the GCC box cis-acting element in vitro and in yeast.The down-regulated expression level of AaERF1 evidently reduced the resistance to B. cinerea in A. annua.The overall results showed that AaERF1 positively regulated the resistance to B. cinerea in A. annua.

View Article: PubMed Central - PubMed

Affiliation: Fudan-SJTU-Nottingham Plant Biotechnology R&D Center, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, People's Republic of China.

ABSTRACT
Plants are sessile organisms, and they can not move away under abiotic or biotic stresses. Thus plants have evolved a set of genes that response to adverse environment to modulate gene expression. In this study, we characterized and functionally studied an ERF transcription factor from Artemisia annua, AaERF1, which plays an important role in biotic stress responses. The AaERF1 promoter had been cloned and GUS staining results of AaERF1 promoter-GUS transgenic A. annua showed that AaERF1 is expressed ubiquitiously in all organs. Several putative cis-acting elements such as W-box, TGA-box and Py-rich element, which are involved in defense responsiveness, are present in the promoter. The expression of AaERF1 can be induced vigorously by methyl jasmonate as well as by ethephon and wounding, implying that AaERF1 may activate some of the defense genes via the jasmonic acid and ethylene signaling pathways of A. annua. The results of electrophoretic mobility shift assay (EMSA) and yeast one-hybrid experiments showed that AaERF1 was able to bind to the GCC box cis-acting element in vitro and in yeast. Ectopic expression of AaERF1 could enhance the expression levels of the defense marker genes PLANT DEFENSIN1.2 (PDF1.2) and BASIC CHITINASE (ChiB), and increase the resistance to Botrytis cinerea in the 35S::AaERF1 transgenic Arabidopsis. The down-regulated expression level of AaERF1 evidently reduced the resistance to B. cinerea in A. annua. The overall results showed that AaERF1 positively regulated the resistance to B. cinerea in A. annua.

Show MeSH
Related in: MedlinePlus