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Engineering rice with Trichoderma chitinase gene for the management of sheath blight disease

Trichoderma viride strain, MNT7 produced chitinases which were subjected to differential precipitation with ammonium sulphate and SDS-PAGE. A42 kDa protein from the 0-40% ammonium sulphate precipitation (ASP) fractions and a 30 kDa protein from the 40-80% ASP fractions were subjected to chitin affinity romatography and were immunologically detected by western blots using anti-barley chitinase antiserum. These chitinases were found to exhibit antifungal activity against the rice sheath blight pathogenic fungus,Rhizocotonia solani (Strain, RS7). N-terminal amino acid sequencing of these two proteins were performed and the N-terminal sequence of the 42KDa chitinase exhibited a very high homology of fungal chitinase hither to reported, while the 30kDa chitinase appeared to have no sequence homology to any of the chitinase reported up till now. The MNT7 42 k Da chitinase characterized in the present study appears to be the first of its kind to be reported from the biocontrol fungus, T. viride.

To strengthen the defensive capabilities into rice that mare naturally active in the plant, an effort has been made to introduce tlp (thaumatin like protein) and chi 11 (chitinase) genes in to rice cultivars namely Taipei309, EYI105 (japonica cultivars), ASD16, IR50, Basmati 370 and White Ponni (indica cultivars)using Biolistic method.

Four putative transgenic lines of Taipei 309 and a single putative transgenic line of White Ponni were recovered with tlp gene. Those evolved putative transgenic lines of taipei 309 and White Ponni were proved to be positive in PCR and southern for the presence of hph gene and also it was found that an induction of a new protein of size (23 kDa) was confirmed by Western analysis. EYI105 variety with chi11 ,one putative transgenic line proved to be positive to histochemical GUS assay. 
 

Source

Agriculture Sprayers and Dusters by ASPEE Group Of Companies, India. www.aspee.com

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This article is written by Ms. R. Swarnambigai of Tamil Nadu Agricultural University, Coimbatore on behalf of ASPEE Agriculture Research and Development Foundation. ASPEE Group of Companies is the leading manufacturer and exporter of Agriculture Sprayers and Dusters

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