Expression of a Chitinase Gene from Metarhizium anisopliae in Tobacco Plants Confers Resistance against Rhizoctonia solani

Abstract

The chit1 gene from the entomopathogenic fungus Metarhizium anisopliae, encoding the endochitinase CHIT42, was placed under the control of the CaMV 35S promoter, and the resulting construct was transferred to tobacco. Seventeen kanamycin-resistant transgenic lines were recovered, and the presence of the transgene was confirmed by polymerase chain reactions and Southern blot hybridization. The number of chit1 copies was determined to be varying from one to four. Copy number had observable effects neither on plant growth nor development. Substantial heterogeneity concerning production of the recombinant chitinase, and both general and specific chitinolytic activities were detected in leaf extracts from primary transformants. The highest chitinase activities were found in plants harboring two copies of chit1 inserts at different loci. Progeny derived from self-pollination of the primary transgenics revealed a stable inheritance pattern, with transgene segregation following a mendelian dihybrid ratio. Two selected plants expressing high levels of CHIT42 were consistently resistant to the soilborne pathogen Rhizoctonia solani, suggesting a direct relationship between enzyme activity and reduction of foliar area affected by fungal lesions. To date, this is the first report of resistance to fungal attack in plants mediated by a recombinant chitinase from an entomopathogenic and acaricide fungus.

Heterologous Expression of Transaldolase Gene Tal from Saccharomyces cerevisiae in Fusarium oxysporum for Enhanced Bioethanol Production

“…6). This CaMV35S promoter has most commonly been used in plant transformation studies [42]. It has also conferred strong gene expression in different fungi, for example, Ganoderma lucidum and Pleurotus citrinopileatus [43].…”

Secondary metabolite gene clusters in the entomopathogen fungus Metarhizium anisopliae: genome identification and patterns of expression in a cuticle infection model

“…Metarhizium spp. are models for host-pathogen interaction studies and virulence factor discovery [2–4] as well as for the development of potential novel applications [5–7]. Additionally, this genus comprises unique evolutionary traits, harboring well-characterized transitional species with varying degrees of host specificity.…”

Clonal variation in Scots pine (Pinus sylvestris) and in transgenic silver birch (Betula pendula)

“…Transferred chitinase genes originate either from plants (Lin et al 1995, Kishimoto et al 2004, Chye et al 2005, Takashaki et al 2005, Tohidfar et al 2005, Vellicce et al 2006, Xiao et al 2007, He et al 2008, fungi (Terakawa et al 1997, Mora and Earle 2001, Kumar et al 2009, Kern et al 2010, Prasad et al 2012 or viruses (Corrado et al 2008). …”