Résumé
Aims@#Metarhizium anisopliae is an entomopathogenic fungus (EPF) that exists naturally in the environment and potentially be used as a biological control agent against many insect pests. This study aims to evaluate the effect of nutrient additives on the yield and viability of M. anisopliae spore and to determine the optimum incubation period for maximum spore production.@*Methodology and results@# In this study, M. anisopliae was cultivated by solid-state fermentation using rice as a growth medium. Three different nutrient additives were examined which aimed to maximize the production of M. anisopliae spores. Among the three nutrient additives evaluated, yeast (1.84 ± 0.04 g) supported better growth and spore production than molasses (0.58 ± 0.04 g) and palm oil (0.47 ± 0.09 g). The incubation period between 2-6 weeks produced higher spore yield (0.97 ± 0.02 g spores) at week 4 with a better spore viability (86.30 ± 0.45%) at week 2. @*Conclusion, significance and impact of study@#Hence, it is suggested that the optimum incubation period is between 2 and 6 weeks after inoculation, and M. anisopliae could be mass produced in large quantities on rice substrate with the addition of yeast as the nutrient additives.
Sujets)
Agents de lutte biologique , Viabilité microbienne , MetarhiziumRésumé
Insecticidal nematodes or entomopathogenic nematodes (EPNs) in the genera Heterorhabditis and Steinernema are lethal obligate parasites of insect pests. These nematodes carry specific pathogenic bacteria which are released into the insect hemocoel after penetration of the insect host. The economic importance of entomopathogenic nematodes (EPNs) is increasing as the nematodes are amenable for mass production, formulation, handling and application on a large scale and is currently marketed worldwide for use. The objective of the review is to discuss the principles of formulation and quality control, latest development and future perspectives of the EPNs formulation for the successful use of EPNs as bio insecticides.
Résumé
Black Sigatoka disease caused by Mycosphaerella fijiensis is the most severe banana disease worldwide. The pathogen is in an invasive phase in Brazil and is already present in most States of the country. The potential of 29 isolates of Trichoderma spp. was studied for the control of black Sigatoka disease under field conditions. Four isolates were able to significantly reduce disease severity and were further tested in a second field experiment. Isolate 2.047 showed the best results in both field experiments and was selected for fungicide sensitivity tests and mass production. This isolate was identified as Trichoderma atroviride by sequencing fragments of the ITS region of the rDNA and tef-1α of the RNA polymerase. Trichoderma atroviride was as effective as the fungicide Azoxystrobin, which is recommended for controlling black Sigatoka. This biocontrol agent has potential to control the disease and may be scaled-up for field applications on rice-based solid fermentation.
A Sigatoka-negra causada por Mycosphaerella fijiensis é a doença mais destrutiva da bananeira em termos mundiais. O patógeno está em uma fase invasiva no Brasil e já se encontra distribuído na maior parte dos Estados do país. O potencial de 29 isolados de Trichoderma spp. para o controle da Sigatoka-negra foi estudado sob condições de campo. Quatro isolados foram capazes de reduzir significativamente a severidade da doença e foram selecionados para um segundo experimento de campo. O isolado 2.047 apresentou os melhores resultados e foi utilizado em testes de sensibilidade a fungicidas e produção massal. Esse isolado foi identificado como Trichoderma atroviride por meio do sequenciamento de fragmentos da regiões ITS do rDNA e tef-1α da RNA polymerase. Trichoderma atroviride foi tão efetivo no controle da Sigatoka-negra quanto o fungicida Azoxystrobin, que é recomendado para o controle da doença. O agente de controle biológico tem potencial para o controle da Sigatoka-negra e pode ser produzido em massa em arroz autoclavado para aplicações no campo.
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The plant parasitic nematodes infect the root tissues of the plant causing root galls that lead to reduced water and mineral uptake in the plant root system. Nematophagous fungus are used as biocontrol for the nematodes. Among those Nematoctonus are one of the species used as bioagent. Nematoctonus species produces an extensive mycelium and capture many nematodes with hour glass shaped adhesive knobs on the hyphae. Nematodes become attached to these adhesive knobs and the cuticle of nematode is penetrated by the infective hyphae. This isolate of Nematoctonus robustus is characterized by hyaline mycelium, dikaryotic in nature containing genetically two different nuclei in each cell, having distinct clamp connection. The fungus has better colonizing ability on natural solid substrates like wheat straw and rice straw. It also show good ability to colonize on different cereal grains and various other waste products like coconut coir and FYM etc. This species is one of the best used for mass production and effective for control of plant parasitic nematodes.
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Recently, the Q biotype of tobacco whitefly has been recognized as the most hazardous strain of Bemisia tabaci worldwide, because of its increased resistance to some insecticide groups. As an alternative control agent, we selected an Isaria javanica isolate as a candidate for the development of a mycopesticide against the Q biotype of sweet potato whitefly. To select optimal mass production media for solid-state fermentation, we compared the production yield and virulence of conidia between 2 substrates (barley and brown rice), and we also compared the effects of various additives on conidia production and virulence. Barley was a better substrate for conidia production, producing 3.43 x 10(10) conidia/g, compared with 3.05 x 10(10) conidia/g for brown rice. The addition of 2% CaCO3 + 2% CaSO4 to barley significantly increased conidia production. Addition of yeast extract, casein, or gluten also improved conidia production on barley. Gluten addition (3% and 1.32%) to brown rice improved conidia production by 14 and 6 times, respectively, relative to brown rice without additives. Conidia cultivated on barley produced a mortality rate of 62% in the sweet potato whitefly after 4-day treatment, compared with 53% for conidia cultivated on brown rice. The amendment of solid substrate cultivation with additives changed the virulence of the conidia produced; the median lethal time (LT50) was shorter for conidia produced on barley and brown rice with added yeast extract (1.32% and 3%, respectively), KNO3 (0.6% and 1%), or gluten (1.32% and 3%) compared with conidia produced on substrates without additives.
Sujets)
Caséines , Fermentation , Glutens , Hemiptera , Hordeum , Ipomoea batatas , Mortalité , Spores fongiques , Nicotiana , Virulence , LevuresRésumé
New method for rearing Spodoptera frugiperda in laboratory shows that larval cannibalism is not obligatory. Here we show, for the first time, that larvae of the fall armyworm (FAW), Spodoptera frugiperda (Lepidoptera, Noctuidae), can be successfully reared in a cohort-based manner with virtually no cannibalism. FAW larvae were reared since the second instar to pupation in rectangular plastic containers containing 40 individuals with a surprisingly ca. 90% larval survivorship. Adult females from the cohort-based method showed fecundity similar to that already reported on literature for larvae reared individually, and fertility higher than 99%, with the advantage of combining economy of time, space and material resources. These findings suggest that the factors affecting cannibalism of FAW larvae in laboratory rearings need to be reevaluated, whilst the new technique also show potential to increase the efficiency of both small and mass FAW rearings.
Résumé
Clamidósporos são estruturas de sobrevivência do fungo nematófago Pochonia chlamydosporia. Objetivou-se, com este trabalho, avaliar diferentes substratos, teores de água e formas de inóculo para a produção in vitro de clamidósporos de P. chlamydosporia. Inicialmente, testaram-se os substratos grãos de milho triturado, grãos de arroz e casca de café e os tipos de inóculo meio líquido concentrado ou diluído (1:40) e discos de cultura, colonizados por P. chlamydosporia. Posteriormente, testou-se o substrato grãos de milho triturado suplementado ou não com caldo de batata com os inóculos do fungo nas formas de discos de micélio ou suspensão aquosa. As maiores produções de clamidósporos g-1 de substrato foram obtidas nos substratos grãos de milho triturado e casca de café, e as melhores formas de inóculo foram meio líquido diluído (1:40) e discos de micélio. A suplementação do substrato grãos de milho triturado com caldo de batata não aumentou a produção de clamidósporos do fungo e a melhor forma de inóculo do fungo foi a de discos de micélio. Além disso, testaram-se os substratos canjica, grãos de arroz, casca de café e fibra de coco, umedecidos com diferentes quantidades de água; o substrato casca de café umedecido por diferentes períodos; a casca de café enriquecida ou não com sacarose e farinha de arroz e o substrato grãos de arroz esterilizado no forno micro-ondas com diferentes quantidades de água. Apenas o substrato grãos de arroz, em todas as quantidades de água testadas, apresentou maior produção de clamidósporos. Não houve diferença na produção de clamidósporos no substrato casca de café umedecido por diferentes períodos e apenas quando a casca de café foi enriquecida com farinha de arroz, apresentou maior média de clamidósporos g-1 de substrato. Todos os tratamentos apresentaram grande produção de clamidósporos g-1 quando o substrato utilizado foi o de grãos de arroz tratados no forno de micro-ondas. O melhor meio de cultivo de P. chlamydosporia para a produção de clamidósporos foi o substrato contendo grãos de arroz.
Chlamydospores are survival structures of the nematophagous fungus Pochonia chlamydosporia. The objective of this study was to evaluate different substrates, different contents of water and kinds of inoculum for the production of Pochonia chlamydosporia chlamydospores. The substrates evaluated were: milled maize, rice grains and coffee husk; all of them were inoculated with disks of culture, concentrated liquid media or diluted liquid media (1:40) colonized by P. chlamydosporia. Besides, the substrates milled maize, supplemented or not with potato broth, and inoculated with fungus disks or aqueous suspension were evaluated. Milled maize and coffee husk were the best substrates for chlamydospores production. The best inoculum kinds were disks of culture and diluted liquid media (1:40) colonized by P. chlamydosporia. The supplementation of milled maize with potato broth did not improve the production of chlamydospores, and the best inoculum form was disks of fungus. Moreover, it was also studied the substrates rice grains, coffee husk and coconut fibers with different rates of water; the substrate coffee husk moistened by different periods; the supplementation of coffee husk with rice flour or dextrose; and the substrate grains of rice sterilized in microwave oven with different rates of water. The substrate grains of rice, in all of the amounts of water tested showed increase in production of chlamydospores. There was no difference in the chlamydospores production of the coffee husk substrate, when moistened by different periods of time and only when it was supplemented with rice flour it showed higher mean of clamydospores per gram of substrate. All treatments evaluated showed higher production of chlamydospores in the substrate grains of rice treated in the microwave oven. The best substrate for the production of chlamydospores of P. chlamydosporia was the rice grains.
Résumé
O objetivo deste trabalho foi avaliar o crescimento de cultivares de morangueiro em sistema de cultivo orgânico, observando a influência da posição da planta no canteiro sobre a produção acumulada e a partição de massa seca entre os órgãos aéreos das plantas. O experimento foi realizado na Embrapa Clima Temperado, Estação Experimental Cascata, Pelotas, RS. Os tratamentos constaram da combinação de dois fatores experimentais: cultivar ('Albion', 'Aromas', 'Camarosa' e 'Camino Real') e a posição das plantas no canteiro (Central e Bordadura). O delineamento experimental adotado foi o de blocos completos casualizados com parcelas divididas e quatro repetições. A cultivar 'Camarosa' apresentou maior produção de massa seca dos órgãos aéreos das plantas (62,3; 27,5 e 88,0g planta-1 para folha, coroa e frutos, respectivamente) e, juntamente com a 'Aromas', maior produtividade (64,2 e 59,1Mg ha-1, respectivamente). A cultivar 'Aromas' apresentou a maior contribuição proporcional dos frutos para a composição do total de massa seca da planta (60,6 por cento) e o maior índice de colheita (88 por cento). A posição das plantas não influenciou o crescimento e a produtividade, indicando ausência de efeito de bordadura para as cultivares 'Albion', 'Aromas', 'Camarosa' e 'Camino Real'.
The aim of this research was to evaluate the growth of strawberry cultivars growing in organic crop system observing the influence of plant position in the bed on accumulated dry matter production and its partitioning among the aboveground plant organs. The experiment was conducted in Embrapa Clima Temperado, Estação Experimental Cascata, Pelotas, RS. The treatments were compose by the combination of two experimental factors: cultivar ('Aromas', 'Albion', 'Camarosa' and 'Camino Real') and plant position in the bed (Central and Border). The experimental design used was randomized blocks with split-plots and four replications. 'Camarosa' cultivar presented highest dry mass production of aboveground organs (62.3; 27.5 and 88.0g plant-1 for leaves, crown and fruits, respectively), and together 'Aromas', highest yield (64.2 and 59.1Mg ha-1, respectively). 'Aromas' cultivar presented highest fruit dry mass contribution for the total aboveground dry mass production (60.6 percent) and the highest harvest index (88 percent). The plant position in the bed did not influence the growth and yield of the plants, which indicate no border effect for the 'Albion', 'Aromas', 'Camarosa' and 'Camino Real' strawberry cultivars.
Résumé
Using the tissue culture technology to achieve the fast propagation of P.ternata and through the photosynthetic characters and yield comparison,two high-yield indivaduals were obtained. The result showed that the coefficent of propagation can get to 1∶6.23 per generation, through 3 month culture the average diameter of the in vitro tuber can get 0.88 cm. P.ternata was a typical shade-plant, of which LCP was only 700?mol?photons/s?m2. Photo-inhibition was observed among all the three leaf-types of P.ternata, and the willow leaf type showed the most obvious photo-inhibition. Both of the light response Pn curve and single tuber weight were significant different among leaf styles. Result for studying the chlorophyll fluorescence parameters showed that all the difference of NPQ, qP and qN among three leaf-types of P.ternata were significant and the difference of potential-photosynthetic productivity among them was significant. The willow leaf style had the strongest light-tolerance capacity.The individuals of T2(peach-leaf type) had the highest Pn and single tuber weight among 11 superior individuals and the individuals of L2(willow-leaf type) showed the strongest light-tolerance capacity. Through 6 month propagation ten thousands in virto tuber of T2 and L2 each were gotten. And after growing in Wenzhou experimental base for one year, the total gross weight of the high-yield individual tuber get to 100kg each.
Résumé
Platelet activating factor (PAF; 1-O-alkyl-2-acetyl-sn-glycero-3-phosphocholine) is a potent lipid mediator in a variety of physiological events. PAF is also involved in various pathological events including allergy and inflammation. PAF-acetylhydrolase (PAF-AH) hydrolyzes PAF to produce inactive lyso-PAF. Thus, overproduction of PAF-AF will be useful for the therapeutic valuation of the enzyme. In this study, we established an overproduction method of bovine PAF-AH in Escherichia coli system. We used bovine mammary gland for cDNA cloning. The cDNA had two mismatches of amino acid sequences (Thr-247 to Met and Ile-431 to Thr) compared with the previously reported PAF-AH cDNA (bovine spleen, NM_174578). The recombinant PAF-AH of 43 kDa in molecular size reacted with human PAF-AH polyclonal antibody and showed a strong PAF-AH enzyme activity in an in vitro assay system. The recombinant PAF-AH produced by this study can be applied for various experiments including in vivo models to test its protective activity against PAF-related diseases.