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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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Sequence of AaERF1 promoter region and construction of AaERF1 promoter-GUS vector.(A) The sequence of AaERF1 promoter region. The transcription initiation site and the translation start site are in bold, underlined letters. Numbers indicate the position relative to the transcription start site. Putative cis-acting regulatory elements involved in defense responsiveness in AaERF1 promoter are in bold. (B) Construction of AaERF1 promoter-GUS vector. PstI and EcoRI are the enzymes used in the construction.
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pone-0057657-g001: Sequence of AaERF1 promoter region and construction of AaERF1 promoter-GUS vector.(A) The sequence of AaERF1 promoter region. The transcription initiation site and the translation start site are in bold, underlined letters. Numbers indicate the position relative to the transcription start site. Putative cis-acting regulatory elements involved in defense responsiveness in AaERF1 promoter are in bold. (B) Construction of AaERF1 promoter-GUS vector. PstI and EcoRI are the enzymes used in the construction.

Mentions: The promoter sequence of AaERF1(JQ513909)was cloned by genomic walking (Figure 1A). To observe the expression pattern of AaERF1 in details, the AaERF1 promoter was subcloned to the pCAMBIA1391Z vector (Figure 1B) and then AaERF1 promoter-GUS transgenic A. annua plants were generated. Six lines of the transgenic A. annua plants expressing the GUS and three lines for the wild-type background were prepared. All the lines showed similar fusion protein expression. GUS activity was detected in all tissues examined, including roots, stems, leaves and flowers (Figure 2A, 2B, 2C and 2D). In 1-month-old plants, GUS activity was high in root tips, stems and leaves (Figure 2A, 2B and 2C). During the flowering period, GUS activity was also detected in flower buds. So, AaERF1 is ubiquitously expressed in A. annua. From Figure 2B and 2C, GUS expression was also detected in the glandular trichomes and T-shaped trichomes. No signals were observed in the negative control plants transformed with pCAMBIA1391 empty vector (Figure S1).


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)

Sequence of AaERF1 promoter region and construction of AaERF1 promoter-GUS vector.(A) The sequence of AaERF1 promoter region. The transcription initiation site and the translation start site are in bold, underlined letters. Numbers indicate the position relative to the transcription start site. Putative cis-acting regulatory elements involved in defense responsiveness in AaERF1 promoter are in bold. (B) Construction of AaERF1 promoter-GUS vector. PstI and EcoRI are the enzymes used in the construction.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0057657-g001: Sequence of AaERF1 promoter region and construction of AaERF1 promoter-GUS vector.(A) The sequence of AaERF1 promoter region. The transcription initiation site and the translation start site are in bold, underlined letters. Numbers indicate the position relative to the transcription start site. Putative cis-acting regulatory elements involved in defense responsiveness in AaERF1 promoter are in bold. (B) Construction of AaERF1 promoter-GUS vector. PstI and EcoRI are the enzymes used in the construction.
Mentions: The promoter sequence of AaERF1(JQ513909)was cloned by genomic walking (Figure 1A). To observe the expression pattern of AaERF1 in details, the AaERF1 promoter was subcloned to the pCAMBIA1391Z vector (Figure 1B) and then AaERF1 promoter-GUS transgenic A. annua plants were generated. Six lines of the transgenic A. annua plants expressing the GUS and three lines for the wild-type background were prepared. All the lines showed similar fusion protein expression. GUS activity was detected in all tissues examined, including roots, stems, leaves and flowers (Figure 2A, 2B, 2C and 2D). In 1-month-old plants, GUS activity was high in root tips, stems and leaves (Figure 2A, 2B and 2C). During the flowering period, GUS activity was also detected in flower buds. So, AaERF1 is ubiquitously expressed in A. annua. From Figure 2B and 2C, GUS expression was also detected in the glandular trichomes and T-shaped trichomes. No signals were observed in the negative control plants transformed with pCAMBIA1391 empty vector (Figure S1).

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