Enhancing the pathogenicity of Arthrobotrys conoides and A. oligospora against Meloidogyne javanica J2 by transferring of protease (AcI) gene and evaluation of antagonistic capability of transgenic isolates

Publication date: Available online 31 March 2018 Source:Biological Control Author(s): Seyedeh Lavin Nourani, Ebrahim Mohammadi Goltapeh, Naser Safaie, Mokhtar Jalali Javaran, Ebrahim Pourjam Antagonistic fungi are well-known as viable alternatives to chemical control of root-knot nematodes. In this paper, serine protease Ac1 as an important pathogenicity factor was used to enhance the antagonistic activity of Arthrobotrys conoides and A. oligospora against Meloidogyne javanica J2 (second stage juveniles). Ac1 gene was extracted from A. conoides and cloned in pCAMBIA1304 vector. The recombinant plasmid was then transferred to these fungi using two strains of Agrobacterium tumefaciens (LBA4404 and AGL1). Transgenic isolates were confirmed by PCR amplification of Hygromycin B resistance gene (hph) as selectable marker, protease assay using casein substrate and in vitro bioassay. Transferring of Ac1 gene by homologous recombination to A. conoides, increased protease activity. Our results showed that production of protease in both transgenic species was increased compared to wild type. Bioassay results indicated that pathogenicity rates of transformants, in both the number of traps formed and the number of trapped J2, increased compared to the wild type. The results showed that inhibition of root-knot nematode, M. javanica, was markedly increased by transgenic isolates of Arthrobotrys spp. compared to the wild types. Graphical abstract

Source: Biological Control - March 31, 2018 Category: Biology Source Type: research