J.R. Montealegre, F. Ochoa, X. Besoain, R. Herrera, and L.M. Pérez. 2014. In vitro and glasshouse biocontrol of Rhizoctonia solani with improved strains of Trichoderma spp. Cien. Inv. Agr. 41(2):197-206. The potential of Trichoderma spp. fusants for the biocontrol of Rhizoctonia solani was compared with the ability of their corresponding parental strains. Their effect was tested in vitro using two R. solani strains, 509 (AG 2-1) and 618 (AG 4). The highest inhibitions in growth in dual cultures were obtained with the ThF2-1 (89.79%), ThF3-3 (90.55%), ThF4-15 (91.75%) and ThF5-8 (77.67%) fusants on R. solani 509; only ThF2-1 was able to inhibit the growth of R. solani 618 (60.19%). The inhibitory effect on growth was mainly due to diffusible metabolites. Percent mortality and canker level in tomato plants were evaluated in glasshouse experiments where all of the evaluated fusants suppressed plant mortality, but only ThF2-1 and ThF5-8 significantly decreased the canker level.

J.R. Montealegre, F. Ochoa, X. Besoain, R. Herrera y L.M. Pérez. 2014. Biocontrol de Rhizoctonia solani in vivo y en invernadero, con cepas mejoradas de Trichoderma spp. Cien. Inv. Agr. 41(2):197-206. Se comparó la capacidad de cepas de Trichoderma spp., obtenidas previamente por fusión de protoplastos, con la de sus correspondientes cepas parentales, para biocontrolar a Rhizoctonia solani. El efecto biocontrolador se analizó sobre dos cepas de R. solani: 509 (GA 2-1) y 618 (GA 4). Las cepas producto de fusión de protoplastos de Trichoderma spp. fueron más efectivas que al menos una de las correspondientes cepas parentales. La inhibición más alta en experimentos de cultivos duales, se observó con las cepas ThF2-1 (89,79%), ThF3-3 (90,55%), ThF4-15 (91,75%) y ThF5-8 (77,67%) sobre R. solani 509; mientras que el efecto biocontrolador sobre R. solani 618, sólo logró un 60,19% de inhibición con la cepa ThF2-1. El efecto inhibidor del desarrollo se debió principalmente a la secreción de metabolitos difusibles. El porcentaje de mortalidad y nivel de cancro en plantas de tomate se evaluó en experimentos de invernadero, en los que todas las cepas producto de la fusión de protoplastos de Trichoderma spp. suprimieron la mortalidad de plantas de tomate, y solamente ThF2-1 y ThF5-8 disminuyeron significativamente el nivel de cancro.

Biorend SC (quitosana), BC-1000 EC (extrato de toranja mais flavonóides), e ECO-100 SC (bioflavonóides mais ácidos orgânicos, fitoalexinas cítricas, ácidos graxos glicerídeos e açúcares) inibiram em 80,1%, 79,0% e 76,5%, respectivamente, a podridão causada por Botrytis cinerea quando utilizados no tratamento pós-colheita de frutos de maçã sob condições controladas.Tratamento combinado de Biorend SC, com aplicação tanto em pré como no pós-colheita, proporcionou 49,9% de inibição da podridão, enquanto BC-1000 e ECO-100 EC não foram efetivos. Nenhum desses produtos inibiu a podridão cinzenta, quando utilizados em tratamento de pré-colheita em condições controladas ou em tratamento de pós-colheita em condições comerciais. O polissacarídeo algal ulvana, utilizado nos tratamentos de pós-colheita, reduziu em 56% a podridão cinzenta das maçãs em condições controladas, mas não teve efeito inibitório nos tratamentos combinados de pré e pós-colheita. A incapacidade dos produtos em ativar mecanismos de defesa (quitinases e peroxidases) nos frutos, após o tratamento em pré-colheita, foi consistente com a falta de controle da podridão nesse tipo de ensaio. Pelos resultados, sugere-se que os produtos naturais utilizados apresentam potencial para a utilização no manejo integrado da podridão de Botrytis quando aplicados em pós-colheita.

Biorend SC (chitosan), BC-1000 EC (grapefruit extract plus bioflavonoids) and ECO-100 SC (bioflavonoids plus organic acids, citric phytoalexins, fatty acids, glycerides and sugars), respectively, suppressed grey rot of apple caused by B. cinerea by 80.1%, 79.0% and 76.5% when used as post-harvest treatments under controlled conditions. When applied as combined pre- and post-harvest treatments Biorend SC inhibited fruit rot by 49.9 %, while BC-1000 EC and ECO-100 SC were ineffective. None of the products inhibited fruit rot when applied as pre-harvest treatments under controlled conditions or as post-harvest treatments under commercial conditions. The algal polysaccharide ulvan used in post-harvest treatments suppressed grey rot by 56.0% under controlled conditions, but had no inhibitory effect on combined pre- and post-harvest treatments. The inability of products to activate defense mechanisms (chitinase and peroxidase) of fruits was consistent with the unsuccessful control of rot by pre-harvest treatment. The results suggest that the natural products used have potential for use in integrated management of Botrytis rot when applied after harvest.

The obtainment of 30 new strains from native Trichoderma harzianum after UV light irradiation (UV-A and UV-C), and of 82 strains resulted from protoplast fusion were accomplished. The new strains, initially selected for their growing rate under low temperature and high pH conditions, as well as for their innocuousness on tomato plants, were tested for in vitro inhibition of Pyrenochaeta lycopersici in dual cultures and due to secretion of volatile and diffusible metabolites. All the UV-A and UV-C selected candidate mutants were innocuous to tomato plants, but none of them showed improvement in their biocontrol activity on P. lycopersici. Th12A20.1 increased 1.3 and 1.9 fold the total fresh weight of Fortaleza tomato plants when compared to its parental strains Th12 and Th11, respectively. The selected candidate mutants obtained through protoplast fusion were also innocuous to tomato plants, but only ThF1-2 and ThF4-4 inhibited 1.3 fold (in dual cultures) and 5 fold (due to secretion of volatile metabolites) the growth of P. lycopersici, respectively, in relation to the mean inhibitory effect of both parents. Therefore, these candidate mutants could be included in experiments under field conditions.

Trichoderma harzianum 650 (Th650) and Paenebacillus lentimorbus 629 (Pl629) selected earlier for their ability to control Rhizoctonia solani, Fusarium solani and F. oxysporum in vitro, were applied alone or combined with solarization (summer assay) and/or with methyl bromide (MeBr) (summer and winter assays) to a soil with a high inoculum level, for the control of tomato root rot caused by the complex F. oxysporum f. sp. lycopersici - Pyrenochaeta lycopersici - Rhizoctonia solani. Evaluations were also performed independently for root damage caused by P. lycopersici, and also for R. solani in the summer assay. MeBr decreased tomato root damage caused by the complex from 88.7% to 21.2% and from 78.4% to 35.7% in the summer and in the winter assay, respectively. None of the bio-controllers could replace MeBr in the winter assay, but Th650 and Pl629 reduced root damage caused by this complex in the summer assay. Treatments with bio-controllers were improved by their combination with solarization in this season. Independent evaluations showed that the positive control of Th650 towards R. solani and the lack of effect on P. lycopersici correlates well with the endochitinase pattern expressed by Th650 in response to these phytopathogens. Root damage caused by R. solani can be controlled at a similar level as it does MeBr in summer assays, thus representing an alternative to the use of this chemical fungicide for the control of this phytopathogen

Bacteria from the rhizoplane and surrounding soil of healthy and Rhizoctonia solani diseased tomato plants, cropped in greenhouse of the V Region of Chile, were collected. The best bacterial strains, based on their ability to control development of three R. solani isolates (identified as belonging to the anastomosis groups AG-2-1, AG‑4), were identified as B. subtilis (one isolate) and B. lentimorbus (two different isolates). All bacterial isolates resulted effective for the in vitro control of growth of all R. solani isolates, where the control mechanisms used by the bacteria do not involve the secretion of fungal cell wall hydrolytic enzymes. R. solani AG-2-1 was more sensitive than R. solani AG-4. On the other hand, all bacteria grew well in conditions similar to those that can be found at the field level (considering pH, salinity, Fe3+ and temperature) and showed a good capacity of tomato root colonization. These results suggest that the B. subtilis and B. lentimorbus isolates studied have an excellent potential to be used as biocontrol agents of R. solani in tomato greenhouses at the field level.