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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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Expression patterns of AaERF1 in response to hormone and stress treatments by RT-Q-PCR.A. Relative expression levels of AaERF1 after MeJA (100 µM). B. Relative expression levels of AaERF1 after ethephon (500 µM). C. Relative expression levels of AaERF1 after wound treatment. Total RNA was isolated respectively from A. annua leaves under different treatments for different periods of time (0 h, 0.5 h, 1 h, 3 h, 6 h, 12 h and 24 h) followed by RT-Q-PCR analysis with the gene-specific primers AaERF1-RT-F and AaERF1-RT-R. Values indicate the mean fold relative to sample 0 h. Actin is used as a control for normalization. Data are averages ± SE from three independent experiments.
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pone-0057657-g003: Expression patterns of AaERF1 in response to hormone and stress treatments by RT-Q-PCR.A. Relative expression levels of AaERF1 after MeJA (100 µM). B. Relative expression levels of AaERF1 after ethephon (500 µM). C. Relative expression levels of AaERF1 after wound treatment. Total RNA was isolated respectively from A. annua leaves under different treatments for different periods of time (0 h, 0.5 h, 1 h, 3 h, 6 h, 12 h and 24 h) followed by RT-Q-PCR analysis with the gene-specific primers AaERF1-RT-F and AaERF1-RT-R. Values indicate the mean fold relative to sample 0 h. Actin is used as a control for normalization. Data are averages ± SE from three independent experiments.

Mentions: In this study, RT-Q-PCR analysis was used to obtain the expression pattern of AaERF1 after hormone and stress treatments including MeJA (100 µM), ethephon (500 µM) and wound treatments. The transcript level of AaERF1 was increased rapidly and peaked within 1 h after MeJA treatment, followed by a gradually decline (Figure 3A). The treatment with ethephon shows a similar expression pattern with the treatment of MeJA (Figure 3B). The transcript level of AaERF1 was also sensitive to stress treatments. Wounding could induce a significant accumulation of AaERF1 transcript in a short time period (0.5 h). Then the transcript level was quickly decreased (Figure 3C). 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)

Expression patterns of AaERF1 in response to hormone and stress treatments by RT-Q-PCR.A. Relative expression levels of AaERF1 after MeJA (100 µM). B. Relative expression levels of AaERF1 after ethephon (500 µM). C. Relative expression levels of AaERF1 after wound treatment. Total RNA was isolated respectively from A. annua leaves under different treatments for different periods of time (0 h, 0.5 h, 1 h, 3 h, 6 h, 12 h and 24 h) followed by RT-Q-PCR analysis with the gene-specific primers AaERF1-RT-F and AaERF1-RT-R. Values indicate the mean fold relative to sample 0 h. 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-g003: Expression patterns of AaERF1 in response to hormone and stress treatments by RT-Q-PCR.A. Relative expression levels of AaERF1 after MeJA (100 µM). B. Relative expression levels of AaERF1 after ethephon (500 µM). C. Relative expression levels of AaERF1 after wound treatment. Total RNA was isolated respectively from A. annua leaves under different treatments for different periods of time (0 h, 0.5 h, 1 h, 3 h, 6 h, 12 h and 24 h) followed by RT-Q-PCR analysis with the gene-specific primers AaERF1-RT-F and AaERF1-RT-R. Values indicate the mean fold relative to sample 0 h. Actin is used as a control for normalization. Data are averages ± SE from three independent experiments.
Mentions: In this study, RT-Q-PCR analysis was used to obtain the expression pattern of AaERF1 after hormone and stress treatments including MeJA (100 µM), ethephon (500 µM) and wound treatments. The transcript level of AaERF1 was increased rapidly and peaked within 1 h after MeJA treatment, followed by a gradually decline (Figure 3A). The treatment with ethephon shows a similar expression pattern with the treatment of MeJA (Figure 3B). The transcript level of AaERF1 was also sensitive to stress treatments. Wounding could induce a significant accumulation of AaERF1 transcript in a short time period (0.5 h). Then the transcript level was quickly decreased (Figure 3C). 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