Chitinases protect and defend plants against infection, especially fungal one. Chitinase identification and purification protocols have to consider methods' income and outcome. The study suggested a modified zymogram that enabled enzyme identification and purification. Reliability of the suggested protocol was documented by immunoblotting pattern of the purified chitinases. Also, biological activity of the purified chitinase was assured by its hydrolytic and antifungal activities. The modified protocol enabled separation of both constitutive and induced chitinases from supper marmand tomato cultivar that was challenged with Alternaria solani. Constitutive expression of chitinase was identified as 26 and 27 kDa while facultative expression of chitinase that induced by A. solani inoculation was detected at 30 kDa. Activity assessment indicated that the purified 30 kDa was differentially expressed as an induced protein to contribute in plant defense. Chitinase of 30 kDa has much more chitinolytic and antifungal activities than the constitutive one that already expressed for protection. Sequencing of the purified 30 kDa documented its belonging to family Chi9 gene.
Key words: Antifungal activity, Chitinlytic activity, Constitutive expression, Induced expression, Pathogenesis Related proteins (PRs)
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