Heterologous expression of the Hsp24 from Trichoderma asperellum improves antifungal ability of Populus transformant Pdpap-Hsp24 s to Cytospora chrysosperma and Alternaria alternate
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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.
KeywordsPopulus Trichoderma asperellum Heat shock protein Plant transgene Antifungus
This work was supported by grants from the National High Technology Research and Development Program (863 Program) (2013AA102701) and the Natural Science foundation of Heilongjiang Province of China (QC2011C003) and the Fundamental Research Funds for the Central Universities (2572014CA11).
- Barcala M, Garcia A, Cubas P, Almoguera C, Jordano J, Fenoll C, Escobar C (2008) Distinct heat-shock element arrangements that mediate the heat shock, but not the late-embryogenesis induction of small heat-shock proteins, correlate with promoter activation in root-knot nematode feeding cells. Plant Mol Biol 66:151–164CrossRefPubMedGoogle Scholar
- Boter M, Amigues B, Peart J, Breuer C, Kadota Y, Casais C, Moore G, Kleanthous C, Ochsenbein F, Shirasu K, Guerois R (2007) Structural and functional analysis of SGT1 reveals that its interaction with HSP90 is required for the accumulation of Rx, an R protein involved in plant immunity. Plant Cell 19:3791–3804CrossRefPubMedPubMedCentralGoogle Scholar
- Duan Y, Jiang YZ, Ye SL, Karim A, Ling ZY, He YQ, Yang SQ, Luo KM (2015) PtrWRKY73, a salicylic acid-inducible poplar WRKY transcription factor, is involved in disease resistance in Arabidopsis thaliana. PLANT CELL REP 34(5) 831–841Google Scholar
- Hanhong B (2011) Trichoderma Species as Abiotic and Biotic Stress Quenchers in Plants. Res J Biotechnol 6(3):73–79Google Scholar
- Liu ZH, Yang XX, Sun DM, Song JZ, Chen G, Juba O, Yang Q (2010) Expressed sequence tags-based identification of genes in a biocontrol strain Trichoderma asperellum. Mol Biol 37:3673–3681Google Scholar
- Montero-Barrientos M, Hermosa R, E.Cardoza R, SantiagoGutierrez, Carlos Nicolas, EnriqueMonte (2010) Transgenic expression of the Trichoderma harzianum hsp70 gene increases Arabidopsis resistance to heat and other abiotic stresses. J Plant Physiol 167:659–665Google Scholar