Molecular cloning of chitinase 33 (chit33) gene from Trichoderma atroviride

Abstract

In this study Trichoderma atroviride was selected as over producer of chitinase enzyme among 30 different isolates of Trichoderma sp. on the basis of chitinase specific activity. From this isolate the genomic and cDNA clones encoding chit33 have been isolated and sequenced. Comparison of genomic and cDNA sequences for defining gene structure indicates that this gene contains three short introns and also an open reading frame coding for a protein of 321 amino acids. The deduced amino acid sequence includes a 19 aa putative signal peptide. Homology between this sequence and other reported Trichoderma Chit33 proteins are discussed. The coding sequence of chit33 gene was cloned in pEt26b(+) expression vector and expressed in E. coli.

Impact of Trichoderma spp. on soybean seed germination and potential antagonistic effect on Sclerotinia sclerotiorum

“…Nowadays, numerous commercial biofungicides, such as Bio-Fungus (Belgium), Trichodex (Israel), Trieco (India), Supresivit (U.S., Europe), Promot (California), Binab T (U.K., Sweden), Root Shield (USA, Europe), etc., are based on Trichoderma viride, T. harzianum and T. koningii strains that are efficient biocontrol agents of several phytopathogenic fungi (https://attra.ncat.org/attra-pub/summaries/summary.php?pub=146). So far, Trichoderma species have been mainly registered as effective antagonists of soilborne pathogens, such as Sclerotinia sclerotiorum (Mansour et al, 2008;Matroudi et al, 2009;Ibarra-Mediná et al, 2010), S. minor (Jones and Stewart, 1997;Ibarra-Mediná et al, 2010), Macrophomina phaseolina (Ilyas et al, 1985;Larralde-Corona et al, 2008), Rhizoctonia solani (Maisura and Patel, 2009;da Silva et al, 2012), Fusarium spp. (Sempere and Santamarina, 2009;Zang and Wang, 2012), Pythium spp.…”

The Antagonism Mechanism Of Trichoderma spp. Towards Fusarium solani Mold

“…This mold species suggested can produce the highest β-1,3 glucanase enzyme compared with another Trichoderma species [7]. Trichoderma species are antagonistic to some phytopathogenic fungi because they have the ability to suppress the disease they cause [8]. Trichoderma uses several biocontrol mechanisms such as mycoparasitism, antibiosis, and competition for space and nutrients between the Trichoderma and F. solani in vitro.…”

Transformation of canola by chit33 gene towards improving resistance to Sclerotinia sclerotiorum

“…The DNA fragment containing the chit33 cDNA was isolated from pUCSM2 (Matroudi et al 2008) and subcloned into plant binary expression vector pBI121. Due to the presence of a SacI site in chit33 cDNA, the fragment containing the chit33 open reading frame (ORF) was subcloned into XbaI site of pBI121GUS -(originated from pBI121 vector which has lost its gus gene and SacI site) (Esfahani et al 2012).…”

“…The A. tumefaciens strain LBA4404 harbouring the recombinant binary vector pBITS1 with the chit33 gene was used in the experiments. The chit33-cDNA from T. atroviride was transferred from pUCSM2 (Matroudi et al 2008) …”