The heterologous overexpression of hsp23, a small heat-shock protein gene from Trichoderma virens, confers thermotolerance to T. harzianum.
An EST showing high values of identity with genes coding for small heat shock proteins (sHSPs) was selected from an EST library collection of Trichoderma virens T59. The cDNA gene (hsp23) with a sequence size of 645 bp long was amplified by PCR. The expression of this gene was evaluated in cultures grown at temperatures ranging from 4 to 41 degrees C. An increased level of expression was detected when the fungus was grown at extreme temperatures (4, 10 or 41 degrees C). A high-expression level was also observed when the fungus was grown in 10% ethanol for 4 h. The hsp23 gene was present as a unique copy in the T. virens genome, and a homologous gene was also present in other five investigated Trichoderma species. Strain T. harzianum T34 was transformed with the hsp23 gene from T. virens T59 under the control of the pki (pyruvate kinase) promoter from T. reesei and the ble (phleomycin resistance) gene as selection marker. Statistically significant differences were detected between the strains T34 and two selected transformants in the biomass quantities obtained after heat shock treatment and in the colony diameters after incubation at 4 degrees C for 2 months.
Montero-Barrientos
Marta
M
Centro Hispano-Luso de Investigaciones Agrarias (CIALE), Departamento de Microbiología y Genética, Universidad de Salamanca, Plaza Doctores de la Reina s/n, 37007 Salamanca, Spain.
Cardoza
Rosa E
RE
Gutiérrez
Santiago
S
Monte
Enrique
E
Hermosa
Rosa
R
D000595:Amino Acid Sequence; D015153:Blotting, Western; D015966:Gene Expression Regulation, Fungal; D015723:Gene Library; D006360:Heat-Shock Proteins / Q000235:genetics*; D008969:Molecular Sequence Data; D010692:Phleomycins / Q000494:pharmacology; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011770:Pyruvate Kinase / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000254:growth & development; D017386:Sequence Homology, Amino Acid; D013696:Temperature; D014242:Trichoderma / Q000235:genetics*
D006360:Heat-Shock Proteins; D010692:Phleomycins; D011770:Pyruvate Kinase