Comparison of Effects of Acidic Electrolyzed Water and NaOCl on Tilletia indica Teliospore Germination

Abstract

Definitive identification of free teliospores of Tilletia indica, causal agent of Karnal bunt of wheat, requires polymerase chain reaction (PCR)-based diagnostic tests. Since direct PCR amplification from teliospores has not been reliable, teliospores first must be germinated in order to obtain adequate DNA. We have routinely surface-sterilized teliospores for 2 min with 0.4% (vol/vol) sodium hypochlorite (NaOCl) to stimulate germination and produce axenic cultures. However, we observed that some spores were killed even with a 2-min NaOCl treatment, the shortest feasible duration. Decreasing the NaOCl concentration in our study from 0.4% to 0.3 and 0.2%, respectively, increased teliospore germination, but treatment times longer than 2 min still progressively reduced the germination percentages. In testing alternative methods, we found “acidic electrolyzed water” (AEW), generated by electrolysis of a weak solution of sodium chloride, also surface-sterilized and increased the rate of T. indica teliospore germination. In a representative experiment comparing the two methods, NaOCl (0.4%) for 2 min and AEW for 30 min increased germination from 19% (control) to 41 and 54%, respectively, by 7 days after treatment. Because teliospores can be treated with AEW for up to 2 h with little, if any, loss of viability, compared with 1 to 2 min for NaOCl, treatment with AEW has certain advantages over NaOCl for surface sterilizing and increasing germination of teliospores of suspect T. indica.

Application of electrolyzed water in the food industry

“…EO water can also reduce germination of many fungal species, such as Alternaria spp., Bortrytis spp., Cladosporium spp., Colletotrichum spp., Curvularia lunata, Didymella bryonaie, Epicoccum nigrum, Fusarium spp., Helminthosporium spp., Pestalotia spp., Phomopsis longicolla, Rhodosporidium toruloides, Stagonospora nodorum, Thielaviopsis basicola, Trichoderma spirale, Acidovorax avenae subsp., Erwinia chrysanthemi, Pantoea ananatis, Pseudomonas syringae (Buck, Iersel, Oetting, & Hung, 2002), Aspergillus spp. (Buck et al, 2002;Suzuki et al, 2002b), Botryosphaeria berengeriana (Al-Haq et al, 2002), Monilinia fructicola (Al-Haq et al, 2001;Buck et al, 2002), Penicillium expansum (Okull & Laborde, 2004) and Tilletia indica (Bonde et al, 1999).…”

Observations of the Germination Behavior of <i>Tilletia indica</i> Teliospores on the Soil Surface under Varying Simulated Environmental Conditions

“…The suspension was incubated at 17˚C for 72 hrs; the debris was pelleted by centrifugation at 600×G for 5 min, the supernatant The pellet was suspended by vortex in 0.1% water agar, and 100 μl of the suspension transferred to a 15-ml conical centrifuge tube. Ten milliliters of acidic electrolytic water [11] was added to the 15-ml centrifuge tube and placed on a rocker for 30 min. It was then centrifuge-washed twice for 5 minutes at 600×G, suspended in 300 μl of sterile water, and seeded onto 2% water agar in 10-μl aliquots by the method of Inman et al [20].…”

“…In this study, the direct observation on soil and quantification of germinating teliospores was not determined, but rather the viability of the teliospore recovered from the soil. It should be noted that the process of surface sterilizing the teliospores with acidic electrolytic water [11] prior to seeding water agar dishes has been shown to increase teliospore germination. This limits our conclusion to the observation that over the course of the experimental temperature simulations a portion of the teliospores are capable of germination during the time period of the study.…”

“…to determine the longevity of T. indica survival in soil [10] [11] [12] [13]. Studies were conducted using teliospores mixed in soil contained in micromesh bags.…”

Eficiência de tratamentos alternativos no controle de Pyricularia grisea em sementes de trigo

“…O hipoclorito de sódio já havia sido empregado com sucesso no controle de diferentes patógenos como os de pós-colheita Botrytis cinerea, Mucor piriformis e Penicillium expansum em peras (Spotts & Peters, 1980), na germinação de patógenos da parte aérea, Tilletia indica, em trigo (Bonde et al, 1999), no tratamento de sementes de brássicas para Xanthomonas campestris pv. campestris (Babadoost et al, 1996), de arroz para bactérias e fungos (Chun et al, 1997), de ornamentais para Nimbya gomphrenae, Stemphylium vesicarium e Alternaria tagetica (Wu et al, 2001) e de tubérculos de batata para Rhizoctonia solani e Streptomyces scabies (Errampalli & Johnston, 2001).…”