title: Heterologous expression of the Hsp24 from Trichoderma asperellum improves antifungal ability of Populus transformant Pdpap-Hsp24 s to Cytospora chrysosperma and Alternaria alternate.
abstract: The tolerance of plants to biotic and abiotic stresses could be improved by transforming with fungal resistance-related genes. In this study, the cDNA sequence (GenBank Acc. No. KP337939) of the resistance-related gene Hsp24 encoding the 24 kD heat shock protein was obtained from the biocontrol fungus Trichoderma asperellum ACCC30536. The promoter region of Hsp24 contained many cis-regulators related to stresses response, such as "GCN4" and "GCR1" etc. Hsp24 transcription in T. asperellum was up-regulated under six different environmental stresses, compared with the control. Furthermore, following heterologous transformation into Populus davidiana × P. alba var. Pyramidalis (Pdpap), Hsp24 was successfully transcribed in transformant Pdpap-Hsp24s. Pathogen-related genes (PRs) in four Pdpap-Hsp24s were up-regulated compared with those in the control Pdpap (Pdpap-Con). After co-culture of Pdpap-Hsp24s with the weak parasite Cytospora chrysosperma, the transcription of genes related to hormone signal pathway (JA and SA) were up-regulated in Pdpap-Hsp24s, and ethidium bromide (EtBr) and Nitro-blue tetrazolium (NBT) staining assays indicated that the cell membrane permeability and the active oxygen content of Pdpap-Hsp24s leaves were lower than that of the control Pdpap-Con. And when the Pdpap-Hsp24s were under the Alternaria alternate stress, the activities of superoxide dismutase (SOD) and peroxidase (POD) got higher in Pdpap-Hsp24s than that in Pdpap-Con, and the disease spots in Pdpap-Con leaves were obviously larger than those in Pdpap-Hsp24s leaves. In summary, Hsp24 of T. asperellum ACCC30536 is an important defense response gene, and its heterologous expression improved the resistance of transformant Pdpap-Hsp24s to C. chrysosperma and A. alternate.
authors.affiliation: School of Forestry, Northeast Forestry University, 26 Hexing Road, 150040, Harbin, China.
authors.forename: S D
authors.initials: SD
authors.lastname: Ji
authors.affiliation: School of Forestry, Northeast Forestry University, 26 Hexing Road, 150040, Harbin, China.
authors.forename: Z Y
authors.initials: ZY
authors.lastname: Wang
authors.affiliation: School of Forestry, Northeast Forestry University, 26 Hexing Road, 150040, Harbin, China.
authors.forename: H J
authors.initials: HJ
authors.lastname: Fan
authors.affiliation: The College of Landscape, Northeast Forestry University, 26 Hexing Road, 150040, Harbin, China.
authors.forename: R S
authors.initials: RS
authors.lastname: Zhang
authors.affiliation: School of Forestry, Northeast Forestry University, 26 Hexing Road, 150040, Harbin, China.
authors.forename: Z Y
authors.initials: ZY
authors.lastname: Yu
authors.affiliation: School of Forestry, Northeast Forestry University, 26 Hexing Road, 150040, Harbin, China.
authors.forename: J J
authors.initials: JJ
authors.lastname: Wang
authors.affiliation: School of Forestry, Northeast Forestry University, 26 Hexing Road, 150040, Harbin, China. LZHNEFU@126.com.
authors.forename: Z H
authors.initials: ZH
authors.lastname: Liu
mesh_terms: D000528:Alternaria / Q000235:genetics*; D000935:Antifungal Agents / Q000378:metabolism*; D001203:Ascomycota / Q000235:genetics*; D001483:Base Sequence; D002463:Cell Membrane Permeability; D003001:Cloning, Molecular; D005285:Fermentation / Q000235:genetics; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D015966:Gene Expression Regulation, Fungal; D005800:Genes, Fungal; D009195:Peroxidase / Q000378:metabolism; D030821:Plants, Genetically Modified; D032107:Populus / Q000235:genetics* / Q000382:microbiology; D011401:Promoter Regions, Genetic / Q000235:genetics; D013312:Stress, Physiological / Q000235:genetics; D013482:Superoxide Dismutase / Q000378:metabolism; D014158:Transcription, Genetic; D014170:Transformation, Genetic; D014242:Trichoderma / Q000235:genetics / Q000378:metabolism*
chemical_list: D000935:Antifungal Agents; D005656:Fungal Proteins; D009195:Peroxidase; D013482:Superoxide Dismutase
keywords: Antifungus; Heat shock protein; Plant transgene; Populus; Trichoderma asperellum