Differentially Expressed Peroxidases from Artemisia annua and Their Responses to Various Abiotic Stresses
- 47 Downloads
Artemisia annua is well-known for producing the antimalarial phytomolecule, artemisinin. The role of peroxidases has been hypothesized in artemisinin metabolism owing to the presence of an –O–O– linkage in this sesquiterpene lactone. Earlier, using a microarray, we identified differentially expressed genes, including peroxidases, in plant growth stages having contrasting artemisinin content. Here, three peroxidases—Aa547, having higher expression in low-artemisinin stage, and Aa540 and Aa528, having higher expression in high artemisinin stage, which could be associated with trichomes on the basis of their approximate gene expression pattern inferred from EST counts in UniGene—were selected for full-length cloning, tissue-specific expression profiling, and in silico analyses. The upstream genomic region of Aa547 was cloned and various cis-regulatory elements were identified. All the three candidates were predicted to be class III plant peroxidases. Further, this study aimed to check the responsiveness of the logically selected peroxidase genes to various abiotic stress factors. Taking cues from previous reports and the regulatory elements observed in the Aa547 promoter, hydration, salinity, temperature, salicylic acid, hydrogen peroxide, and methyl jasmonate, were selected to study their effect on the expression of the peroxidase genes through qRT-PCR. The peroxidases were found to be highly sensitive to the various factors but differed in their responses. Broadly, except for responses to high temperature and salicylic acid, the response of Aa547 to various factors was distinct from that of Aa540 and Aa528, which was in line with its distinctness from the other two peroxidases, considering the in planta artemisinin content and predicted structural features.
KeywordsArtemisia annua Peroxidases Phylogenetic analysis Promoter analysis Stress
The authors are grateful to the Director of CSIR-CIMAP, for his encouragement and for providing laboratory facilities. They are also thankful to Dr. Anil K. Gupta, Senior Principal Scientist, CSIR-CIMAP and Curator, National Gene Bank for Medicinal and Aromatic Plants at CSIR-CIMAP, Lucknow, for his help and cooperation. PN thanks CSIR for her Senior Research Fellowship.
This is CIMAP Communication No.: CIMAP/PUB/2017-01.
PN carried out the experiments, recorded the observations, analyzed the data, and helped in manuscript preparation. AK Shasany helped in planning and analysis of data. FK helped in conducting in silico analyses. AK Shukla conceived and planned the study, analyzed the data, and prepared the manuscript.
Compliance with Ethical Standards
Conflict of Interest
The authors declare that they have no conflict of interest.
- Bhakuni RS, Jain DC, Sharma RP, Kumar S (2001) Secondary metabolites of Artemisia annua and their biological activity. Curr Sci 80:35–48Google Scholar
- Cheeseman JM (2007) Hydrogen peroxide and plant stress: a challenging relationship. Plant Stress 1:4–15Google Scholar
- Ferreres F, Figueiredo R, Bettencourt S, Carqueijeiro I, Oliveira J, Gil-Izquierdo A, Pereira DM, Valentão P, Andrade PB, Duarte P, Barceló AR, Sottomayor M (2011) Identification of phenolic compounds in isolated vacuoles of the medicinal plant Catharanthus roseus and their interaction with vacuolar class III peroxidase: an H2O2 affair? J Exp Bot 62:2841–2854CrossRefPubMedGoogle Scholar
- Nair P, Misra A, Singh A, Shukla AK, Gupta MM, Gupta AK, Gupta V, Khanuja SPS, Shasany AK (2013) Differentially expressed genes during contrasting growth stages of Artemisia annua for artemisinin content. PLoS One 8(4):e60375. https://doi.org/10.1371/journal.pone.0060375 CrossRefPubMedPubMedCentralGoogle Scholar
- Narusaka Y, Nakashima K, Shinwari ZK, Sakuma Y, Furihata T, Abe H, Narusaka M, Shinozaki K, Yamaguchi-Shinozaki K (2003) Interaction between two cis-acting elements, ABRE and DRE, in ABA-dependent expression of Arabidopsis rd29A gene in response to dehydration and high-salinity stresses. Plant J 34:137–148CrossRefPubMedGoogle Scholar
- Weisany W, Sohrabi Y, Heidari G, Siosemardeh A, Ghassemi-Golezani K (2012) Changes in antioxidant enzymes activity and plant performance by salinity stress and zinc application in soybean (Glycine max L.) Plant Omics J 5:60–67Google Scholar
- Zhang Y-S, Liu B-Y, Li Z-Q, Ye H-C, Wang H, Li G-F, Han J-L (2004) Molecular cloning of a classical plant peroxidase from Artemisia annua and its effect on the biosynthesis of artemisinin in vitro. Acta Bot Sin 46:1338–1346Google Scholar