Construction of ATMT System for Clonostachys rosea and Its Conditional Optimization of Colonization into Glycyrrhiza uralensis Seeds / 中国实验方剂学杂志

ObjectiveAgrobacterium tumefaciens-mediated transformation （ATMT） of Clonostachys rosea， an endophytic fungus of Glycyrrhiza uralensis seeds， was established and optimized， and orthogonal test was designed to optimize the colonization conditions of C. rosea for G. uralensis seeds， so as to lay foundation for the development of biofertilizer and the breeding of high-quality G. uralensis. MethodThe conditions of ATMT were optimized from three aspects， including the concentration of acetosyringone， co-culture time and the concentration of conidia of recipient fungi. Then， high-quality transformants were selected. Orthogonal test was used to optimize the colonization conditions by taking co-culture temperature， co-culture time and spore concentration as factors and colonization rate as index. ResultWhen spore concentration was 1×107 cfu·mL-1， acetosyringone concentration was 150 μmol·L-1 and the co-culture time was 60 h， the transformation efficiency of C. rosea was the highest， which was 135 transformants per 1×107 recipient fungal spores. The accuracy and stability of the transformations were tested by cloning the marker gene green fluorescent protein （GFP） and β-glucuronidase （GUS） staining. When co-culture temperature was 25 ℃， co-culture time was 36 h and the spore concentration was 1×106 cfu·mL-1， the colonizing rate for C. rosea back dyeing into G. uralensis seeds by seed soaking method was the highest， which was 71.11%. ConclusionThis study successfully establishes stable and efficient technical systems not only of ATMT in C. rosea， but also of colonization of the transformants into G. uralensis seeds， which can lay a foundation for the development of biofertilizer of G. uralensis.

Predatory activity of Butlerius nematodes and nematophagous fungi against Haemonchus contortus infective larvae / Atividade predatória de nematóides Butlerius e fungos nematófagos contra larvas infectantes de Haemonchus contortus

Abstract The purpose of this study was to evaluate the predatory activity of the nematode Butlerius spp. and fungal isolates of Duddingtonia flagrans, Clonostachys rosea, Arthrobotrys musiformis and Trichoderma esau against H. contortus infective larvae (L3) in grass pots. Forty-eight plastic gardening pots containing 140 g of sterile soil were used. Panicum spp. grass seeds (200 mg) were sown into each pot and individually watered with 10 mL of tap water. Twelve days after seeding, the pots were randomly divided into 6 groups (n=8). Two thousand H. contortus infective larvae (L3) were added to each group. Additionally, the following treatments were established: Group 1 – 2000 Butlerius spp. larvae; group 2 – A. musiformis (1x107 conidia); group 3 – T. esau (1x107 conidia); group 4 – C. rosea (1x107 conidia), group 5 – D. flagrans (1x107conidia) and Group 6 – no biological controller (control group). The larval population of H. contortus exposed to Butlerius spp. was reduced by 61.9%. Population reductions of 90.4, 66.7, 61.9 and 85.7% were recorded in the pots containing A. musiformis, T. esau, C. rosea and D. flagrans, respectively. The results of this study indicate that the predatory nematode Butlerius spp. and the assessed fungi display an important predatory activity can be considered suitable potential biological control agents.

Resumo O objetivo deste estudo foi avaliar a atividade predatória do nematoide Butlerius spp. e isolados fúngicos de Duddingtonia flagrans, Clonostachys rosea, Arthrobotrys musiformis e Trichoderma esau contra larvas infectantes (L3) de Haemonchus contortus em vasos plantados com Panicum spp. Foram utilizados quarenta e oito potes plásticos de jardinagem contendo 140 g de solo estéril, 200 mg de sementes de Panicum spp.. Cultivar colonião, foi semeado em cada vaso e, diariamente, molhados com 10 mL de água da torneira. Doze dias após, os vasos foram divididos em 6 grupos (n = 8), e duas mil L3 de H. contortus foram adicionadas a cada vaso. Foram estabelecidos os seguintes tratamentos: Grupo 1 - 2.000 larvas de Butlerius spp.; Grupo 2 - A. musiformis (1x107 conídios); grupo 3 - T. esau (1x107 conídios); grupo 4 - C. rosea (1x107 conídios); grupo 5 - D. flagrans (1x107conidia); e Grupo 6 – somente L3 de H. contortus que serviu como controle negativo. A população de L3 de H. contortus expostas a Butlerius spp. foi reduzida em 61,9%. Redução populacional de 90,4, 66,7, 61,9 e 85,7% foram observadas nos vasos contendo A. musiformis, T. esau, C. rosea e D. flagrans, respectivamente. Os resultados deste estudo indicaram que o nematoide Butlerius spp. e os fungos avaliados exibiram importante atividade predatória e podem ser considerados como agentes de controle biológico.

Insect deterrent and nematicidal screening of microfungi from Mexico and anti-aphid compounds from Gliomastix masseei / Evaluación de la actividad disuasiva de insectos y nematotóxica de microhongos de México y compuestos anti-áfidos de Gliomastix masseei

Fungal metabolites are promising alternatives for the development of biorational pesticides. In this sense, microfungi from tropical regions are valuable sources of natural compounds for pest management. With the aim of broadening the search for new eco-friendly products to manage plant pests, this study was carried out to evaluate the biological activity of 23 tropical fungal extracts on three species of phytophagous insects and a plant parasitic nematode. In addition, the active principles of the most effective extract were identified. The insect deterrent activity of fungal extracts was evaluated on the settling of aphids Myzus persicae and Rhopalosiphum padi, and on the feeding of lepidoptera larva Spodoptera littoralis; the nematostatic activity was evaluated on the mobility of Meloidogyne javanica. Active metabolites from Gliomastix masseei were identified by GC-MS techniques and by comparison with commercial standards. Results showed seven extracts with strong effect on the settling of M. persicae and R. padi (settling inhibition >80%). The calculated median of effective concentration (EC50) values ranged from 8 to 38 µg/cm² for the extracts of Clonostachys rosea and G. masseei, respectively. Bioassay-guided separation of the ethyl acetate extract of G. masseei revealed the presence of fatty acids and their derivatives, where methyl 9-octadecenoate was the most active compound with EC50 values of 16 µg and 35 µg/cm² for M. persicae and R. padi, respectively. Extracts of C. rosea and G. masseei could be a promising option in the control of pest aphids in agriculture.

Los metabolitos fúngicos son agentes prometedores para el desarrollo de plaguicidas biorracionales. En este sentido, los hongos microscópicos de zonas tropicales representan una valiosa fuente de compuestos naturales para el manejo de plagas. Con la finalidad de ampliar la investigación en productos amigables con el medio ambiente, en este estudio se evaluó la actividad biológica de 23 extractos de hongos sobre 3 especies de insectos fitófagos y un nematodo fitoparásito; además se identificaron los componentes del extracto más activo. El efecto disuasivo de los extractos fúngicos se evaluó en el asentamiento de Myzus persicae y Rhopalosiphum padi, así como en la alimentación de Spodoptera littoralis; la actividad nematostática se evaluó sobre la movilidad de Meloidogyne javanica. Los metabolitos activos de Gliomastix masseei se identificaron por cromatografía de gases-espectrometría de masas y por comparación con muestras comerciales. Los resultados mostraron 7 extractos con fuerte efecto en la inhibición del asentamiento (> 80%) de M. persicae y R. padi. Los valores de la concentración efectiva media (CE50) estuvieron en el rango de 8 a 38 µg/cm² para los extractos de Clonostachys rosea y G. masseei, respectivamente. El extracto de acetato de etilo de G. masseei se fraccionó por un proceso biodirigido y reveló la presencia de ácidos grasos y sus derivados, donde el 9-octadecenoato de metilo fue el más activo, con una CE50 de 16 µg/cm² para M. persicae y 35 µg/cm² para R. padi. Los extractos fúngicos de C. rosea y G. masseei pueden ser una alternativa promisoria en el control de áfidos que son plaga en la agricultura.

Screening and Identification of Endophytic Fungi from Schisandra chinensis with Antioxidant Activity / 中国药房

OBJECTIVE:To screen and identify endophytic fungi from Schisandra chinensis with antioxidant activity. METH-ODS:The tissue isolation skill was used to isolate endophytic fungi from roots,leaves,stems and fruits of S. chinensis. And anti-oxidant activity of endophytic fungi were screened by DPPH radical scavenging assay and hydroxyl radical scavenging assay. The to-tal DNA were extracted;the 18S rDNA ITS were amplified and sequenced with primer ITS1 and ITS4;the results of sequencing were analyzed comparatively based on homology to confirm the classification of active strains. RESULTS:23 strains were isolated from S. chinensis. GSR-12,isolated from roots of S. chinensis,had strong antioxidant activities. The scavenging rate on DPPH and the hydroxyl radical were 87.96% and 82.31% respectively. GSR-12 strain was identified as Clonostachys rosea by analyzed com-paratively. CONCLUSIONS:1 strain of C. rosea,isolated from roots of S. chinensis,has strong antioxidant activity.

From the ancient times of the agriculture to the biological control in plants: a little of the history / Dos primórdios da agricultura ao controle biológico em plantas: um pouco de história

The advancement of technology in agriculture driven by the need to increase efficiency in production transformed agriculture in an eminently anti-ecological activity, with large-scale use of industrialized products. Human beans still live the paradigm of agriculture based on the use of chemical inputs, which often brings harmful consequences to the environment. As an alternative to u the use of pesticides to control pests and diseases, biological control is a practice that has been increasingly encouraged. With it, it is possible to think on changes that promote the conservation of natural resources and of the planet humans live in, contributing to strength the new era in biotechnology education.

O avanço da tecnologia na área agrícola, induzido pela necessidade de aumento da eficiência na produção, transformou a agricultura numa atividade eminentemente antiecológica, com uso em larga escala de insumos industrializados. Vivemos ainda o paradigma da agricultura baseada no uso de insumos químicos, a qual muitas vezes traz consequências danosas ao meio ambiente. Como alternativa ao uso de agrotóxicos para o controle de pragas e doenças, o controle biológico é uma prática que vem sendo cada vez mais incentivada. Com ele, pode-se pensar em transformações que, visando a conservação de recursos naturais e do planeta em que vivemos, contribuam para o fortalecimento da nova era biotecnológica em formação.