Three isolates of binucleate Rhizoctonia (BNR) were tested for biological control of damping-off of cucumber seedlings caused by Rhizoctonia solani AG 2-2 and AG 4. BNR isolates L2 (AG Ba) and W1 and W7 (AG A) provided protection of 58 to 71% against virulent isolate C4 of AG 4 and 64 to 75% protection against virulent isolate RH 65 of AG 2-2. Varying protection was provided to the seedlings by the BNR isolates against the virulent R. solani from the two AGs depending on their combination. The BNR isolates did not vary in providing protection to the seedling when tested against virulent C4 when both isolates were inoculated using three different methods, viz. in water agar, combination of water agar and soil and using soil alone. Protection of 58 to 71% was provided by the isolates when inoculation was done on the hypocotyl using water agar, 62.8 to 75% using the combination of water agar and soil, and 75 to 85% when inoculation of both isolates was done in soil. Pre-incubation of BNR W7 or delayed inoculation of C4 (from 0.5 day to longer duration) using the different methods provided an increased protection to the seedlings to give complete inhibition of damping-off disease. Simultaneous inoculation of both BNR W7 and C4 using the three methods failed to provide protection to the seedlings. Among the BNR isolates, BNR W7 showed plant growth promotion in terms of significant increase in plant height (P = 0.0I) and fresh weight (P = 0.05).

Binucleate Rhizoctonia (BNR) W7 was evaluated for the efficacy to control damping-off disease in cucumber seedlings. Results revealed that Lobate appresoria and dome-shaped infection cushion were produced by virulent Rhizoctonia solani C9 AG4 on the surface of the cucumber seedling hypocotyls, causing damping-off, 12 h after inoculation. A significant positive correlation was observed between disease severity rating and number of infection cushions (r = 0.94) and appresoria (r = 0.97) on the hypocotyls of seedlings inoculated with virulent R. solani C9 alone. Inoculation of a nonpathogenic species of BNR prior to virulent R. solani significantly reduced the number of appresoria on hypocotyls surface and inhibited the formation of infection cushions that accompanied by highly significant (P ≤ 0.01) reduction in disease severity in comparison with seedling inoculated with R. solani C9 alone. Formation of mucilaginous material upon recognition of the nonpathogenic isolate of BNR followed by hyphal lysis of BNR W7 6 h after inoculation was responsible for reduction in appresorium formation and complete inhibition of infection cushion formation. However, the number of infection structures produced on cucumber was not significantly different for hypocotyls of seedlings inoculated with R. solani C9 with and without the nonpathogenic BNR W7.

The interaction of binucleate Rhizoctonia (BNR) anastomosis group (AG)-A isolate W7, Rhizoctonia solani AG-4 and cucumber seedlings were investigated to elucidate the mechanism of biocontrol of Rhizoctonia solani by BNR. Hypocotyls of Cucumis sativus L. cv. Jibai were inoculated with a virulent isolate of R. solani AG-4 isolate C4 and examined with light microscopy and scanning and transmission electron microscopy. The hyphae of the virulent isolate colonized the outer surface of the hypocotyl and penetrated the epidermal and cortical cells to the pith of hypocotyls. The hyphae of a non-pathogenic species of BNR isolate W7 did not penetrate the cortical cells but instead colonized the outer epidermal cells 12 h after inoculation. Accumulation of mucilage occurred on the surface after dense colonization of the hypocotyls surface by the BNR. Observation of the hypocotyls surface pre-inoculated with BNR and challenged inoculated with R. solani showed constricted and deformed hyphae of R. solani, which were prevented to penetrate the epidermal cells. Pre-inoculating of BNR isolate caused profused formation of the mucilage which lysed BNR hyphae and restricted the growth of R. solani. The mucilage was identified as pectic substances. Analysis of pectin contents from dried hypocotyls tissues showed highly methylated pectin with low uronic acid contents in the seedlings inoculated with R. solani and unprotected by BNR while BNR-treated seedlings showed less methylated pectin and high amount of uronic acid. Furthermore, the BNR-inoculated seedlings showed significant (P=0.05) increase in calcium, indicating high amount of wall-bound cations in cell walls. Pectic substances accumulation and increased calcium in the cucumber tissues greatly contributed in the protection of cucumber seedlings against Rhizoctonia damping off by a non-pathogenic species of Rhizoctonia.