Molecular cloning of a cytosolic ascorbate peroxidase cDNA from cell cultures of sweet potato and its expression in response to stress.
A cDNA encoding a cytosolic ascorbate peroxidase (APX), swAPX1, was isolated from cell cultures of sweet potato ( Ipomoea batatas) by cDNA library screening, and its expression in the context of various environmental stresses was investigated. swAPX1 contains an ORF of 250 amino acids (27.5 kDa) encoding a protein with a pI value of 5.32. The swAPX1 ORF does not code for a transit peptide, suggesting that the product is a cytosolic isoform. RNA blot analysis showed that swAPX1 gene is expressed in cultured cells and mature leaves, but not in stems, non-storage or storage roots of sweet potato. The level of swAPX1 RNA progressively increased during cell growth in suspension cultures. In leaf tissues, the gene responded differentially to various abiotic stresses, as revealed by RT-PCR analysis. swAPX1 was highly induced in leaves by wounding, and treatment with methyl viologen (50 microM), hydrogen peroxide (440 mM), abscisic acid (ABA; 100 microM) or exposure to high temperature (37 degrees C). In addition, the gene was strongly induced in the leaves following inoculation with a bacterial pathogen ( Pectobacterium chrysanthemi). These results indicate that swAPX1 may be involved in hydrogen peroxide-detoxification and thus help to overcome the oxidative stress induced by abiotic and biotic stresses.
Park
S-Y
SY
Laboratory of Environmental Biotechnology, Korea Research Institute of Bioscience and Biotechnology, Oun-dong 52, Yusong-gu, 305-806 Daejeon, Korea.
Ryu
S-H
SH
Jang
I-C
IC
Kwon
S-Y
SY
Kim
J-G
JG
Kwak
S-S
SS
D000040:Abscisic Acid / Q000494:pharmacology; D000595:Amino Acid Sequence; D060387:Ascorbate Peroxidases; D015152:Blotting, Northern; D002455:Cell Division; D002478:Cells, Cultured; D003001:Cloning, Molecular; D003600:Cytosol; D018076:DNA, Complementary / Q000235:genetics*; D004781:Environmental Exposure; D015971:Gene Expression Regulation, Enzymologic; D018506:Gene Expression Regulation, Plant; D015723:Gene Library; D006540:Herbicides / Q000494:pharmacology; D006861:Hydrogen Peroxide / Q000494:pharmacology; D027723:Ipomoea batatas / Q000201:enzymology* / Q000235:genetics; D008969:Molecular Sequence Data; D016877:Oxidants / Q000494:pharmacology; D018384:Oxidative Stress; D010269:Paraquat / Q000494:pharmacology; D010544:Peroxidases / Q000235:genetics* / Q000378:metabolism; D010937:Plant Growth Regulators / Q000494:pharmacology; D018515:Plant Leaves / Q000166:cytology / Q000378:metabolism*; D018749:RNA, Plant / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid; D013696:Temperature; D014947:Wounds and Injuries
D018076:DNA, Complementary; D006540:Herbicides; D016877:Oxidants; D010937:Plant Growth Regulators; D018749:RNA, Plant; D000040:Abscisic Acid; D006861:Hydrogen Peroxide; D010544:Peroxidases; D060387:Ascorbate Peroxidases; D010269:Paraquat