Chemical and biological control of Sclerotinia stem rot in the soybean crop / Controle químico e biológico da podridão branca da haste na cultura da soja

It was evaluated the effect of fungicides and the microbial control agent Trichoderma harzianum on the inhibition of the carpogenic and ascospore germination of Sclerotinia sclerotiorum. This study also evaluated the chemical, fungicidal and microbial control of white mold or Sclerotinia stem rot of soybean in the field. Three experiments were conducted, as follows: 1) inhibition of carpogenic germination of sclerotia, 2) inhibition of ascospore germination, and 3) control of Sclerotinia stem rot in a soybean crop under field conditions. The treatments evaluated were fluazinam, procymidone, iprodione, thiophanate-methyl, carbendazim, benzalkonium chloride + fluazinam, and T. harzianum. Procymidone resulted in an inhibition of 13.5% and benzalkonium chloride in an inhibition of 13.9% in an ascospore germination test. Fluazinam and procymidone were the most effective in reducing the production of ascospores/apothecium, representing 65.6% and 82.4% of inhibition. Procymidone and fluazinam if combined or not with benzalkonium chloride were the most effective in controlling sclerotinia stem rot under field conditions when applied at the onset of flowering and 15 days later. In the 2009-10 harvest, these two fungicides reduced the incidence of Sclerotinia stem rot by 73.1 and 71.6% and in the 2010-11 harvest by 75.7 and 77.6%, respectively.

Avaliaram-se o efeito de fungicidas e agente de controle biológico Trichoderma harzianum na inibição da germinação carpogênica e de ascósporos de Sclerotinia sclerotiorum. Também foram avaliados os controles químico e biológico do mofo branco ou podridão branca da haste na cultura da soja em condições de campo. Foram desenvolvidos três ensaios, sendo: 1) inibição da germinação carpogênica; 2) inibição da germinação de ascósporos; e 3) controle da podridão branca da haste na cultura da soja em condições de campo. Os tratamentos avaliados foram fluazinam, procimidona, iprodione, tiofanato metílico, carbendazim, cloreto de benzalcônio + fluazinam e T. harzianum. O procimidona resultou em inibição de 13,5% e o cloreto de benzalcônio + fluazinam em 13,9% de inibição no teste de inibição da germinação dos ascósporos. Fluazinam e procimidona foram os mais eficientes na redução da produção de ascósporos/apotécio representando 65,6% e 82,4% de inibição. O procimidona e fluazinam se combinados com cloreto de benzalcônio ou não foram os mais eficientes no controle da podridão branca da haste em condições de campo, quando aplicados no início do florescimento e 15 dias após. Na safra 2009-10, esses dois fungicidas reduziram a incidência da podridão branca da haste, em 73,1 e 71,6% e, na safra 2010-11, reduziram em 75,7 e 77,6%, respectivamente.

Fruiting Body Formation of Cordyceps militaris from Multi-Ascospore Isolates and Their Single Ascospore Progeny Strains

Interest in commercial cultivation and product development of Cordyceps species has shown a recent increase. Due to its biochemical and pharmacological effects, Cordyceps militaris, commonly known as orange caterpillar fungus, is being investigated with great interest. Cultivation of C. militaris has been practiced on a large scale in order to fulfill a demand for scientific investigation and product development. Isolates of C. militaris can be easily established from both spores and tissue. For isolation of spores, ascospores released from mature stromata are trapped in sterile medium. Multi-ascospore isolates, as well as combinations of single ascospore strains, are used for production of fruiting bodies. Progeny ascospore strains can be isolated from artificial fruiting bodies, thus, the cycle of fruiting body production can be continued for a long period of time. In this study, we examined fruiting body production from multi-ascospore isolates and their progeny strains for three generations. F1 progeny strains generally produced a larger number of fruiting bodies, compared with their mother multi-ascospore isolates; however, F2 and F3 progeny strains produced fewer fruiting bodies. Optimum preservation conditions could help to increase the vitality of the progeny strains. In order to retain the fruiting ability of the strains, further testing of various methods of preservation and different methods for isolation should be performed.

Growth of Byssochlamys nivea in pineapple juice under the effect of water activity and ascospore age

The study of thermal resistant mould, including Byssochlamys nivea, is of extreme importance since it has been associated with fruit and fruit products. The aim of this work is to analyze the influence of water activity (a w) and ascospore age (I) on the growth of Byssochlamys nivea in pineapple juice. Mold growth was carried out under different conditions of water activity (a w) (0.99, 0.96, 0.95, 0.93, 0.90) and ascospore age (I) (30, 51, 60, 69, 90 days). Growth parameters as length of adaptation phase (λ), maximum specific growth rate (µmax) and maximum diameter reached by the colony (λ) were obtained through the fit of the Modified Gompertz model to experimental data (measuring radial colony diameter). Statistica 6.0 was used for statistical analyses (significance level α = 0.05). The results obtained clearly showed that water activity is statistically significant and that it influences all growth parameters, while ascospore age does not have any statistically significant influence on growth parameters. Also, these data showed that by increasing a w from 0.90 to 0.99, the λ value substantially decreased, while µmax and λ values rose. The data contributed for the understanding of the behavior of B. nivea in pineapple juice. Therefore, it provided mathematical models that can well predict growth parameters, also helping on microbiological control and products' shelf life determination.

Effect of Preservation Periods and Subcultures on Fruiting Body Formation of Cordyceps militaris In Vitro

Effects of various preservation periods and subcultures on fruiting body formation of Cordyceps militaris were investigated using EFCC C-10995 single ascospore strains. Fruiting body formation by original strains was profuse when preserved at 4degrees C for 5~6 months. Fruiting from subcultures was stable till second to sixth subcultures, after which it decreased sharply. The more the colony color of subcultures changed, the less the fruiting bodies formed. Liquid inoculum preparation of single ascospore strains in the same or separate broths did not affect fruiting body formation. Similarly, two strains C-10995-3 and C-10995-6 in different numbers during liquid inoculum preparation produced similar fruiting bodies.

Genetic Analysis of Pigmentation in Cordyceps militaris

Pigmentation of ascospore-derived isolates from seven different natural specimens of Cordyceps militaris EFCC C-5888, EFCC C-7159, EFCC C-7833, EFCC C-7991, EFCC C-8021, EFCC C-8023 and EFCC C-8179 was observed on the plates of Sabouraud Dextrose agar plus Yeast Extract at 25degrees C under continuous illumination (500 lux). Pigmentation of the wild-type isolates of C. militaris was diverse ranging from yellowish white to orange, while white color was believed as a mutant. Inheritance of pigmentation was found to be controlled by both parental isolates when F1 progeny were analyzed. Pigmentation and mating type were shown to be either independent or distantly linked each other due to the high percentage of non-parental phenotypes among F1 progeny. Crosses between white mutant isolates of C. militaris yielded progeny with wild type pigmentations, indicating that the albino mutations in the parents were unlinked to each other.