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[Role of a protein inhibitor of polygalacturonase in ripening of apples and their resistance of infection with Monilia fructigena, the causative agent of fruit rot].

We investigated the dynamics of the activity of the polygalacturonidase inhibitor protein (PGIP) in apple fruits of six varieties differing in ripening time and correlated it with the degree of damage by the causative agent of fruit rot, Monilia fructigena. The apple varieties studied differed significantly in PGIP activity and degree of damage by Monilia fructigena. The rate of dissemination over fruit tissues was inversely related to PGIP activity. The resistance of apples to M. fructigena increased with ripening. The simultaneous increase in PGIP activity suggests its important role in the reduction of apple damage by fruit rot.

Antifungal Agents↗

Cloning of polygalacturonase (PG) cDNA and inhibition effects of its antisense RNA on the expression of PG gene in transgenic tomato plants.

We have amplified a 1.5-kb PG cDNA including its complete open reading frame, using two primers specific to PG gene and PCR technique. Its fidelity and correctness were confirmed by restriction mapping and partial DNA sequencing. The PG gene was then inserted into a binary vector, pBin437, in an inverted orientation between the 35S promoter with duplicated enhancers and the Nos3' transcriptional termination sequence, and the resulting plasmid pBPG was introduced into LBA 4404. The A. tumefaciens harboring pBPG was used to transform tomato plants. Sixty kanamycin-resistant transformants were selected. PCR detection showed two-third of the transformed plants tested (30 plants) contain the exogenous PG gene. The PG activity assay and SDS-PAGE analysis of proteins from ripening fruit cell wall showed that the PG activity in transgenic tomato plants were inhibited in different extent. One of them, plant 3, gave only 7% PG activity compared with the control (100%). These results suggest that the expression of the anti-PG gene has effectively inhibited the expression of endogenous PG gene. The effect of low-level PG activity on tomato fruit ripening and storage is being investigated.

Base Sequence↗

Effect of various nitrogen salts in beet pulp medium on polygalacturonase activity of Penicillium sp. 7/4B/EI 1 mutant.

The highest PG activity was obtained in beet pulp with (NH4)2HPO4 in amount equivalent to 0.14--0.28% N after 7 days of growth; At optimal concentration, (NH4)2HPO4 did not influence the biomass production, but increased the pH of the culture media to above 6.0. The PG was an extracellular enzyme, acting mainly on highly esterified pectin. The attack was of a random manner, characteristic for the endo-enzymes. The enzyme was active at temperature 20 degrees C--40 degrees C and pH 5.0--6.0.

Culture Media↗