Potential of rhizobacteria native to Argentina for the control of Meloidogyne javanica / Potencial de rizobacterias nativas de Argentina para el control de Meloidogyne javanica

Abstract Biocontrol of the nematode Meloidogyne javanica was studied using the Argentinean strains Pseudomonas fluorescens MME3, TAE4, TAR5 and ZME4 and Bacillus sp. B7S, B9T and B19S. Pseudomonas protegens CHA0 was used as a positive control. Egg hatching and juvenile mortality were evaluated in vitro by exposure of nematodes to bacterial suspensions or their cell-free supernatants (CFS). The effect of bacteria on nematode infestation of lettuce was also studied. results showed that most of the tested strains and CFS reduced egg hatching and juvenile survival in vitro. The bacterial suspension of Bacillus sp. B9T produced the lowest hatching of eggs. Juvenile mortality was higher when M. javanica was exposed to Bacillus sp. than to Pseudomonas spp. suspensions. Except for CFS of B9T, all filtrates inhibited hatching at levels similar to or higher than the biocontrol strain P. protegens CHA0. The CFS of CHA0 showed the highest level of juvenile mortality followed by Bacillus sp. strains and P. fluorescens TAE4. None of the inoculated rhizobacteria reverted the negative effect of infestation on the aerial dry weight of lettuce plants. However, inoculation impacted on reproduction of M. javanica by reducing the development of galls and egg masses on roots and diminishing the number of individuals both on roots and in the substrate, as well as the reproduction factor. These results show that most of the analyzed native strains can control the nematode M. javanica. Among them, P. fluorescens TAE4 and Bacillus sp. B9T showed the most promising performances for the biocontrol of this pathogen and have a potential use in the formulation of commercial products.

Resumen Se estudiaron las cepas argentinas Pseudomonas fluorescens MME3, TAE4, TAR5 y ZME4 y Bacillus sp. B7S, B9T y B19S para el control del nematodo Meloidogyne javanica. Pseudomonas protegens CHA0 se utilizó como control positivo. La eclosión de huevos y la mortalidad de juveniles se evaluaron in vitro al exponerlos a suspensiones bacterianas y a sus sobrenadantes libres de células (SLC). Asimismo, se estudió la inoculación bacteriana sobre la infestación del nematodo en lechuga. Los resultados in vitro indicaron que la mayoría de las cepas, así como sus SLC redujeron la eclosión y la supervivencia de M. javanica. La suspensión de Bacillus sp. B9T produjo los menores niveles de eclosión. La mortalidad de juveniles fue mayor al exponerlos a suspensiones de Bacillus sp. respecto de Pseudomonas spp. Los SLC inhibieron la eclosión de huevos en niveles similares o superiores a P. protegens CHA0, excepto por el de B9T. La exposición a SLC de CHA0 registró la mayor mortalidad, seguido por las cepas de Bacillus sp. y P. fluorescens TAE4. La inoculación bacteriana no revertió el efecto de la infestación sobre el peso seco aéreo de las plantas, sin embargo, afectó la multiplicación de M. javanica lo que redujo el desarrollo de agallas y las masas de huevos, y disminuyó el número de individuos presentes tanto en la raíz como en el sustrato, así como el factor de reproducción. Los resultados indican que la mayoría de las cepas nativas evaluadas son capaces de controlar a M. javanica. Entre ellas, P. fluorescens TAE4 y Bacillus sp. B9T, se presentan como las más promisorias para el control de este patógeno, con potencialidad para ser utilizadas en la formulación de productos biológicos.

Influence of root-knot nematode, Meloidogyne incognita (Kofoid & White) Chitwood infection on different plant growth parameters in Mungbean, Vigna radiata (L.) Wilczek

Root-knot nematode, Meloidogyne incognita (Kofoid & White) Chitwood, is a major threat to mungbean cultivation. The pest causes a significant reduction in plant growth parameters that ultimately results in loss of grain yield. The present study was carried out under glass house condition to study the effect of different inoculum load of root-knot nematode M. incognita on plant growth, nodulation and nematode development and nutrients status of Mungbean. The results revealed a progressive decline in plant growth parameters viz., fresh and dry shoot weight and shoot length with respect to increase in inoculum level. However, fresh and dry root weight showed the opposite trend. The fresh and dry shoot weight was decreased by 44% and 66%, respectively at 4 J2s/g soil. The chlorophyll content in the leaves also decreased with the increase of inoculum level from 100-6000 J2s/pot. Nutrients contents of the plant viz. N, P, K, Ca and Mg were significantly reduced in shoots while in roots these was increased with an increase of inoculum levels. Nodulation was affected by 80% at the highest inoculum level i.e. 6000 J2s/pot. Also leghaemoglobin, bacteroid content and nitrogenase activity was reduced progressively with increased levels of nematode inoculum. Thus, the root-knot nematode, M. incognita interferes with the process of symbiotic nitrogen fixation between mungbean host and rhizobium and that can affect the quality of produce.

Extracts and purified substances of Cabralea canjerana inhibit hatching and extracts of Schinus terebinthifolius kill juveniles of meloidogyne incognita

Nematicidal substances have been identified from plants and are potentially useful for the management of plant-parasitic nematodes. Cabralea canjerana, (Meliaceae) and Schinus terebinthifolius (Anacardiaceae) produce bioactive compounds during their secondary metabolism and little is known about the effect of such substances on plant-parasitic nematodes. In the present study, we assessed the effect of aqueous and ethanolic extracts of C. canjerana and S. terebinthifolius at 1% (m:v) and purified substances from C. canjerana (gedunin, ocotillone, cabraleadiol, a mixture of ocotillone + cabraleadiol and a mixture of shoreic acid + eichlerianic acid) on hatching and mortality of Meloidogyne incognita juveniles. Aqueous extracts of C. canjerana fruits and seeds reduced hatching by 70.3 to 95.7%. Aqueous extracts of S. terebinthifolius fruits killed 42.8 to 77.1% of juveniles. The purified substances of C. canjerana inhibited the hatching of M. incognita from 57 to 90% and did not increase the mortality of juveniles. Therefore, C. canjerana extracts and its purified substances reduce M. incognita hatching and aqueous extracts of S. terebinthifolius kill J2 of this nematode.

Host status of eucalypt species to Pratylenchus brachyurus and Meloidogyne incognita

Plant parasitic nematodes are major threats to Brazilian and world agriculture. Among them, Pratylenchus brachyurus and Meloidogyne incognita stand out as major pests for several crops, including corn, cotton, soybean among others, which can be components of integrated crop-livestock-forestry systems (ICLFs). In this context, information about the host status of eucalypts (Corymbia spp. and Eucalyptus spp.) to plant parasitic nematodes becomes more relevant in Brazil, due to the use of eucalypts in ICLFs. If tree components used on this system increase the population density of P. brachyurus and M. incognita, it is possible that these pathogens could damage shorter-cycle crops. Since information about the host status of eucalypts to plant parasitic nematodes is scarce, this study evaluated the host status of some eucalypt species to P. brachyurus and M. incognita. Two greenhouse trials were done to evaluate the reproduction of P. brachyurus and one to M. incognita, using some of the most cultivated species and hybrids of eucalypts in Brazil. The population growth of P. brachyurus increased on Corymbia citriodora, Eucalyptus grandis, Eucalyptus dunnii × Eucalyptus urophylla, and E. grandis × E. urophylla after ~90 days of inoculation. Conversely, despite M. incognita reproducing well in the control plants, no individuals were recovered from C. citriodora, E. urophylla and E. grandis × E. urophylla, which were classified as resistant plants. Based on both obtained and available data, M. incognita poses no threat to eucalypt species today. However, P. brachyurus is suggested to be a threat to eucalypts.

Parasitism of Meloidogyne exigua races 1 and 2 in coffee plants derived from Timor Hybrid / Parasitismo de Meloidogyne exigua raças 1 e 2 em cafeeiros derivados do Híbrido de Timor

ABSTRACT: To investigate the degree of parasitism of two populations of Meloidogyne exigua, the gall index (GI) and the reproduction factor (RF) of M. exigua races 1 (Est E2) and 2 (Est E1) were analyzed in 47 progenies on F3:4 or F4:5 generation derived from the crossing between Coffea arabica cv. Catuaí Amarelo and Timor Hybrid. C. canephora cv. Apoatã IAC 2258 and C. arabica cv. Catuaí Vermelho IAC 144 were used as resistance and susceptibility checks, respectively. The genotypes that were classified as resistant or susceptible by RF were similarly classified by GI, showing a close relationship between both methodologies. The data also indicated no differences in virulence between the nematode populations, since the progenies showed similar resistance reactions to the M. exigua races 1 and 2. According to GI from the 47 mother plants evaluated, 27 progenies (57.4%) were classified as resistant to M. exigua races 1 and 2, with GI ranging from 0.0 to 1.4 and 20 progenies (42.6%) were susceptible with GI from 2.6 to 4.4. These results showed that most of the evaluated germplasm was very promising in relation to the development of new Arabica coffee cultivars with resistance to M. exigua.

RESUMO: Com o objetivo de investigar o grau de parasitismo de duas populações de Meloidogyne exigua, o índice de galhas (IG) e o fator de reprodução (FR) de M. exigua raças 1 (Est E2) e 2 (Est E1) foram analisados em 47 progênies na geração F3:4 ou F4:5, derivadas do cruzamento entre Coffea arabica cv. Catuaí Amarelo e Híbrido de Timor. Plantas de C. canephora cv. Apoatã IAC 2258 e C. arabica cv. Catuaí Vermelho IAC 144 foram usadas como padrão de resistência e de suscetibilidade, respectivamente. Os genótipos que foram classificados como resistentes ou suscetíveis pelo FR foram similarmente classificados pelo IG, mostrando uma estreita relação entre as duas metodologias para a avaliação da resistência. Os dados também indicaram que não houve diferenças quanto à virulência entre as duas populações do nematoide, uma vez que as progênies mostraram similar reação de resistência a M. exigua raça 1 e 2. De acordo com o IG, das 47 plantas-mãe avaliadas, 27 progenies (57,4%) foram classificadas como resistentes a M. exigua raças 1 e 2, com IG variando de 0,0 a 1,4 e 20 progenies (42,6%) foram suscetíveis, com IG variando de 2,6 a 4,4. Esses resultados mostraram que a maioria dos germoplasmas avaliados foi muito promissora em relação ao desenvolvimento de novas cultivares de café Arábica com resistência a M. exigua.

Immunity of sugarcane cultivars to Meloidogyne enterolobii / Imunidade de cultivares de cana-de-açúcar a Meloidogyne enterolobii

Brazil is currently the world's largest producer and exporter of sugarcane, and the crop has high socioeconomic importance in the country. Root-knot nematodes (Meloidogyne spp.) are one of the major limiting factors in sugarcane production. These plant parasites have wide geographic distribution, high damage potential, and are difficult to control. Recently, the species Meloidogyne enterolobii was identified in sugarcane crops in the state of Pernambuco, Brazil. Given the importance of genetic resistance for integrated nematode management and the lack of research on the M. enterolobii­sugarcane pathosystem, this study aimed to assess the response of sugarcane cultivars to M. enterolobii. Thirteen cultivars were evaluated for their resistance to M. enterolobii based on the nematode reproduction factor. The experiment was conducted in a greenhouse, in a completely randomized design, with 14 replicates. Pre-sprouted sugarcane seedlings were transplanted to 5 L pots, and each pot was considered an experimental unit. At 15 days after transplanting, the seedlings were inoculated with 5,000 eggs and second-stage juveniles of M. enterolobii. Tomato and okra plants were also inoculated to test the viability of the inoculum. At 240 days after inoculation, plant roots were processed and evaluated for nematode number. This parameter was used to calculate the nematode reproduction factor on each cultivar. All sugarcane cultivars were found to be immune to M. enterolobii, with a reproduction factor of 0.

O Brasil é atualmente o maior produtor e exportador mundial de cana-de-açúcar, tendo a cultura grande impacto socioeconômico no país. Dentre os diversos fatores que podem afetar sua produção se destacam os nematoides de galha (Meloidogyne spp.), por apresentarem ampla distribuição geográfica, alto potencial danoso e difícil controle. Recentemente a espécie Meloidogyne enterolobii foi identificada parasitando a cultura da cana-de-açúcar em Pernambuco. Dada à importância da resistência genética no manejo integrado de fitonematoides e a falta de trabalhos envolvendo o estudo desta espécie na cultura, o presente trabalho teve como objetivo avaliar a resposta de cultivares de cana-de-açúcar a M. enterolobii. Foram avaliadas 13 cultivares quanto a sua resistência a esta espécie com base no fator de reprodução do nematoide. O experimento foi conduzido em delineamento inteiramente casualizado, com 14 repetições, e mantido em casa de vegetação. Foram utilizadas mudas pré-brotadas de cana-de-açúcar, transplantadas para vasos de 5L, sendo cada vaso considerado uma unidade experimental. Cada unidade experimental foi inoculada com 5.000 ovos e juvenis de segundo estádio de M. enterolobii 15 dias após os transplantio. Plantas de tomateiro e quiabeiro também foram inoculadas para atestar a viabilidade do inoculo utilizado no experimento. Decorridos 240 dias da inoculação as raízes foram processadas para obtenção da população final e cálculo do fator de reprodução do nematoide de cada tratamento. Todas as cultivares de cana-de-açúcar avaliadas foram resistentes/imunes à M, enterolobii, apresentando fator de reprodução igual a zero.

Development of quantitative detection method for Meloidogyne incognita by qPCR / Desenvolvimento do método de detecção quantitativo para Meloidogyne incognita por qPCR

The root-knot nematode (Meloidogyne spp.) is the most important plant-parasitic nematode genus, they are the most common and destructive pathogens in this group. They produce some of the most drastic symptoms in plants and can significantly reduce the yield of crops. In order to achieve deploy an efficient method of plant-parasitic nematode management, is necessary an identification and quantification accurate and reliable of plant-parasitic nematodes. The aim of this study was to analyze samples in qPCR to detect and quantify M. incognita, in the field samples, comparing different methods of extraction of DNA and its efficacy in establishing the number of individuals. For this purpose the effectiveness of different DNA methods of extraction was compared through the values of CT intervals. For standard curve and method comparisons, we used nematodes multiplied in a greenhouse and carefully separated in the specific quantities of the experiments. For the number of individuals experiment field samples previously counted under an optical microscope were used. The DNA extraction was made from 100 nematodes by the methods: CTAB, Phenol: Chloroform and commercial kit (PureLink® Genomic DNA Kit, Invitrogen). In the comparative analysis using the three methods of DNA extracting from 100 nematodes, it was observed that commercial kit and CTAB methods obtained CT values similar. The CTAB method of extraction, showed less variation in the repeats and greater linearity of standard curve in comparison with other methods tested. So, it was possible to quantify the samples through the CT value intervals, established from different numbers of individuals (1, 10, 25, 100, 250, 500 and 750), in field samples. This study demonstrated that qPCR technique is an alternative sensitive and reliable for the quantification of M. incognita to support laboratories of diagnose and field survey.

Os nematoides-das-galhas (Meloidogyne spp.) é o gênero de fitonematoide mais importante, são os patógenos mais comuns e destrutivos deste grupo. Eles produzem alguns dos sintomas mais drásticos nas plantas e podem reduzir significativamente o rendimento das culturas. Para conseguir implantar um método eficiente de manejo de nematoides parasitas de plantas, é necessária a identificação e quantificação precisa e confiável dos fitonematoides. O objetivo deste estudo foi analisar amostras em qPCR para detectar e quantificar M. incognita, em amostras de campo, comparando diferentes métodos de extração do DNA e sua eficácia no estabelecimento do número de indivíduos. Para este propósito, a eficácia de diferentes métodos de extração de DNA foi comparada através dos valores dos intervalos de Ct. Para comparações padrão de curvas e métodos, usamos nematoides multiplicados em casa de vegetação e cuidadosamente separados nas quantidades específicas dos experimentos. Para o número de indivíduos, foram utilizadas amostras de campo previamente contadas sob um microscópio óptico. A extração de DNA foi realizada a partir de 100 nematoides, pelos métodos: CTAB, Phenol: Clorofórmio e kit comercial (PureLink® Genomic DNA Kit, Invitrogen). Na análise comparativa utilizando os três métodos de extração de DNA a partir de 100 nematoides, observou-se que o kit comercial e os métodos de CTAB obtiveram valores de CT semelhantes. O método de extração CTAB apresentou menor variação nas repetições e maior linearidade da curva padrão em comparação com os demais métodos testados. O coeficiente de correlação (R2) da curva padrão foi de 0,98 indicando uma relação linear entre o valor de Ct e a quantidade de padrões de DNA variando de 90 a 0,00009 ng.µL-1. Assim, foi possível quantificar as amostras através dos intervalos de valores de CT, estabelecidos a partir de diferentes números de indivíduos (1, 10, 25, 100, 250, 500 e 750), em amostras de campo. Este estudo demonstrou que a técnica de qPCR é uma alternativa sensível e confiável para a quantificação de M. incognita, para apoiar laboratórios de diagnóstico e levantamentos de campo.

Influence of Meloidogyne incognita race 1 on the development of clones of Coffea canephora, variety "Jequitibá Incaper 8122" / Influência de Meloidogyne incognita raça 1 no desenvolvimento de clones de Coffea canephora, variedade "Jequitibá Incaper 8122"

Root-knot nematode is one of the most important phytosanitary problems for Conilon coffee, as it reduces productivity and is difficult to handle. We aimed at studying the infectivity and damage caused by M. incognita race 1 in the "Jequitibá Incaper 8122" intermediate maturity coffee variety. The experiment was conducted in a greenhouse, in completely randomized design, with five replicates. The clones composing the variety "Jequitibá Incaper 8122" were inoculated with 2,000 eggs + second-stage juveniles of M. incognita race 1. Uninoculated plants were the control. Evaluations were performed 180 days after inoculation, considering the plant height (H), stem diameter (SD), number of leaves (NOL), leaf area (LA), number of plagiotropic branches (NPB), number of nodes (NN), chlorophyll content (CHLO), shoot dry matter (SDM), root fresh matter (RFM), final population (FNP), and reproduction factor (NRF). The nematode reduced NOL in clones 208 and 209, NRF in clones 201, 203, 207 and 208, NN in clones 203, 207, 208 and 209, CHLO in clones 201, 204, 206, 207 and 209, SDM in clones 201, 203, 204 and 205 and RFM in clones 205 and 207. M. incognita race 1 FNP and NRF were larger in clones 208, 201, 207 and 203. Clone 202 had FNP and NRF equal to zero, being immune to the nematode. Clone 206 presented the lowest NRF value among clones parasitized by M. incognita.(AU)

O nematoide-das-galhas é um dos mais importantes problemas fitossanitários para o cafeeiro conilon, por reduzir a produtividade e ser de difícil manejo. Objetivou-se estudar a infectividade e os danos causados por M. incognita raça 1 na variedade de café conilon de maturação intermediária "Jequitibá Incaper 8122". O experimento foi conduzido em casa de vegetação, em DIC, com cinco repetições. Os clones que compõem a variedade "Jequitibá Incaper 8122" foram inoculados com 2.000 ovos + juvenis de segundo estádio de M. incognita raça 1. Plantas não inoculadas constituíram a testemunha. As avaliações foram realizadas 180 dias após a inoculação, sendo avaliados: altura da planta (ALT), diâmetro do caule (DCA), número de folhas (NFO), área foliar (AFO), número de ramos plagiotrópicos (NRP), número de nós (NN), teor de clorofila (CLO), massa seca da parte aérea (MSA), matéria fresca da raiz (MFR), população final (PFN) e fator de reprodução (FRE). O nematoide reduziu o NFO nos clones 208 e 209, NRP nos clones 201, 203, 207 e 208, NN nos clones 203, 207, 208 e 209, CLO nos clones 201, 204, 206, 207 e 209, MSA nos clones 201, 203, 204 e 205 e MFR nos clones 205 e 207. PFN e FRE de M. incognita raça 1 foram maiores nos clones 208, 201, 207 e 203; o clone 202 teve a PFN e a FRE igual a zero, apresentando-se imune ao nematoide. O clone 206 apresentou o menor valor de FRE entre os clones parasitados por M. incognita.(AU)

Effect of Shea Nut Shell Biochar on Root Knot Nematodes (Meloidogyne spp.) of Tomato (Solanum lycopersicum L.)

Effect of shea nut shell biochar on root knot nematodes and performance of tomato was investigated under nematode inoculated soils. Steam sterilized soil was admixed with biochar, which was later inoculated with 1000 second stage juveniles (J2) two weeks after transplanting. Tomato variety (Petomech-GH) was planted in potting medium of soil to biochar ratio of one part of biochar (250 g) is to one part of soil (1B1S), one part of biochar is to two parts of soil (1B2S), two parts of biochar is to one part of soil (2B1S), and no biochar application (control). Steam sterilized soil amended with biochar inoculated with 1000 second stage juveniles (J2). The result indicated that, biochar increased the pH of the soil, lessened the adverse effects of Meloidogyne spp., resulting in decline in galling and improvement in growth and yield of tomato. Increased biochar concentration resulted in decreased nematode gall formation on the roots of the tomato plant. Biochar amended soils resulted in lower egg masses. Increased biochar concentration resulted in decreased performance of tomato plant. Tomato plants treated with low biochar concentrations (1B2S and 1B1S) produced higher fruit numbers and weights, and plant biomass.

Nematicidal potential of essential oils of Ageratum fastigiatum, Callistemon viminalis and Schinus terebinthifolius / Potencial nematicida de óleos essenciais de Ageratum fastigiatum, Callistemon viminalis e Schinus terebinthifolius

Some essential oils are rich in nematicidal compounds and can be used for the management of plant-parasitic nematodes. Laboratory and field studies aiming to evaluate the efficiency of this compounds are necessary. The objective of this research was to assess the nematicidal potential of essential oils from leaves of Ageratum fastigiatum and Callistemon viminalis, and green and mature fruits of Schinus terebinthifolius on the root-knot nematode Meloidogyne javanica. In laboratory, the essential oil of S. terebinthifolius green fruits reduced hatching by more than 80% and increased juvenile mortality by 300% when compared to Tween 20 + water. In the field, none of the essential oils controlled M. javanica in lettuce. In conclusion, the essential oil of S. terebinthifolius green fruits reduces the egg hatching and kills J2 of M. javanica in laboratory. In an infested field with an average of 555 J2/100 cm3 of soil, the application of the essential oils of S. terebinthifolius, C. viminalis and A. fastigiatum does not control M. javanica in lettuce.

Alguns óleos essenciais são ricos em compostos nematicidas e poderiam ser usados no manejo de fitonematoides. Estudos laboratoriais e de campo que visam avaliar a eficiência desses compostos são necessários. O objetivo desta pesquisa foi avaliar o potencial nematicida de óleos essenciais de folhas de Ageratum fastigiatum e Callistemon viminalis e frutos verdes e maduros de Schinus terebinthifolius sobre o nematoide de galhas Meloidogyne javanica. Em laboratório, o óleo essencial de frutos verdes de S. terebinthifolius reduziu mais de 80% a eclosão e aumentou em 300% a mortalidade de juvenis em comparação com Tween 20 + água. No campo, nenhum óleo essencial controlou M. javanica em alface. Em conclusão, o óleo de frutos verdes de S. terebinthifolius reduz a eclosão e mata J2 de M. javanica em laboratório. Em campo com infestação média de 555 J2/100 cm3 de solo, a aplicação dos óleos essenciais de S. terebinthifolius, C. viminalis e A. fastigiatum não controla M. javanica em alface.

Evaluation of root-knot nematode disease control and plant growth promotion potential of biofertilizer Ning shield on Trichosanthes kirilowii in the field

Abstract Biofertilizer Ning shield was composed of different strains of plant growth promotion bacteria. In this study, the plant growth promotion and root-knot nematode disease control potential on Trichosanthes kirilowii in the field were evaluated. The application of Ning shield significantly reduced the diseases severity caused by Meloidogyne incognita, the biocontrol efficacy could reached up to 51.08%. Ning shield could also promote the growth of T. kirilowii in the field by increasing seedling emergence, height and the root weight. The results showed that the Ning shield could enhance the production yield up to 36.26%. Ning shield could also promote the plant growth by increasing the contents of available nitrogen, phosphorus, potassium and organic matter, and increasing the contents of leaf chlorophyll and carotenoid pigment. Moreover, Ning shield could efficiently enhance the medicinal compositions of Trichosanthes, referring to the polysaccharides and trichosanthin. Therefore, Ning shield is a promising biofertilizer, which can offer beneficial effects to T. kirilowii growers, including the plant growth promotion, the biological control of root-knot disease and enhancement of the yield and the medicinal quality.

Nematicidal Compounds from the Leaves of Schinus terebinthifolius Against Root-knot Nematode, Meloidogyne incognita Infecting Tomato

The root-knot nematode, Meloidogyne incognita caused a serious damage to many plants. The phenolic components of the leaves of Schinus terebinthifolius were investigated as potential nematicidal agents for M. incognita. Nine compounds were isolated and characterized as viz., 1,2,3,4,6-pentagalloyl glucose (1), kaempferol-3-O-α-L-rhamnoside (Afzelin) (2), quercetin-3-O-α-L-rhamnoside (Quercetrin) (3), myricetin (4), myricetin-3-O-α-L-rhamnoside (Myricetrin) (5), methylgallate (6), protocatechuic acid (7), quercetin (8), and gallic acid (9) using nuclear magnetic resonance (NMR) spectroscopy. Compound 1 showed pronounced nematicidal activity compared to Oxamyl as a positive control. It showed the lowest eggs-hatchability (34%) and the highest mortality in nematode population (21% after 72 hours of treatment) at a concentration of 200 µg/mL. It exhibited the best suppressed total nematode population, root galling and number of eggmasses in infected tomato plants. The total carbohydrates and proteins were also significantly induced by 1 with reduction in total phenolics and increase in defense-related proteins. Thus, compound 1 could be a promising, more safe and effective natural nematicidal agent for the control of root-knot nematodes.

Caenorhabditis elegans as an indicator of toxicity of Bacillus thuringiensis strains to Meloidogyne incognita race 3 / Caenorhabditis elegans como indicador de toxicidade de estirpes de Bacillus thuringiensis a Meloidogyne incognita raça 3

ABSTRACT: The cotton plant (Gossypium hirsutum) is affected by several diseases of economic importance, among them root-knot nematode (Meloidogyne incognita races 3 and 4). Methods to control this disease include the application of nematicides, solarization, deep plowing, crop rotation and use of antagonistic microorganisms. Among species of Bacillus, there are strains that act as bioregulators and antagonists of several pathogens. Tests to identify these strains are hampered by the difficulty of obtaining large populations of the pathogen and by the time of execution of the in vivo tests that should be conducted for about 90 days. The objective of this research was to compare the toxicity of B. thuringiensis strains to two nematodes, M. incognita and Caenorhabditis elegans, evaluating the possibility of using C. elegans as an indicator for the selection of strains with biocontrol potential against M. incognita. Therefore, the toxicity of nine B. thuringiensis strains on C. elegans and M. incognita was evaluated under laboratory and greenhouse conditions. Most strains toxic to C. elegans in vitro were also toxic to M. incognita, and three of them (S906, S1192, S2036) significantly reduced the populations of the two nematodes. The toxic effect of B. thuringiensis strains on C. elegans was like that reported for the same bacterial isolates on M. incognita in vivo. These results suggested that it is plausible to use C. elegans as an indicator of toxicity for selection of B. thuringiensis strains toxic to M. incognita.

RESUMO: O algodoeiro (Gossypium hirsutum) é acometido por várias doenças de importância econômica, dentre as quais a meloidoginose (Meloidogyne incognita raças 3 e 4). Entre os métodos de controle dessa doença, destacam-se as aplicações de nematicidas, a solarização, a aração profunda, a rotação de culturas e o uso de microrganismos antagonistas. Dentre as espécies do gênero Bacillus, existem estirpes que atuam como biorreguladores e antagonistas de vários patógenos. Os testes para identificação dessas estirpes são prejudicados pela dificuldade de se obter grandes populações do patógeno e pelo tempo de execução dos testes in vivo que devem ser conduzidos por cerca de 90 dias. Diante disso, o presente trabalho teve como objetivo comparar a toxicidade de estirpes de B. thuringiensis a dois nematoides, M. icognita e Caenorhabditis elegans, verificando a possibilidade de empregar C. elegans como indicador para a seleção de estirpes com potencial de biocontrole contra M. incognita. Para tanto, a toxicidade de nove estirpes de B. thuringiensis para C. elegans e M. incognita foi avaliada em laboratório e em casa de vegetação. A maioria das estirpes tóxicas ao C. elegans in vitro, também foi tóxica ao M. incognita, sendo que três delas (S906, S1192, S2036) reduziram significativamente as populações dos dois nematoides. O efeito tóxico apresentado pelas estirpes de B. thuringiensis contra C. elegans foram similares aos apresentados pelos mesmos isolados contra M. incognita in vivo. Esses resultados sugerem que é plausível o uso do C. elegans como indicador de toxicidade para seleção de estirpes de B. thuringiensis tóxicas a M. incognita.

Morphological, biochemical, and molecular characterization of Meloidogyne spp. populations from Brazilian soybean production regions / Caracterização morfológica, bioquímica e molecular de populações de Meloidogyne spp. de regiões brasileiras produtoras de soja

ABSTRACT: Soybean is a commodity of great economic importance worldwide, particularly in Brazil, world’s second largest producer. Nematodes, especially those of the Meloidogyne genus, severely limit productivity. Identification of nematode species is important for effective soybean management. Here, 26 populations of root-knot nematode (Meloidogyne spp.) from 15 municipalities in the states of Bahia, Mato Grosso, Goias, and Minas Gerais were characterized based on the morphology of the female perineal region, esterase profile, and identification based on amplification of specific regions of the population genome. Among the Meloidogyne spp. populations obtained, M. incognita and M. javanica, were identified. No mixed populations were present in the samples. Diagnosis based on molecular analysis was shown to be reliable and the fastest for characterization of nematode populations compared to other methods analyzed.

RESUMO: A soja é uma commodity de grande importância econômica em todo mundo, especialmente no Brasil, segundo maior produtor mundial. Os nematoides, em especial os do gênero Meloidogyne, causam grandes limitações na produtividade. A identificação das espécies de nematoides é uma informação importante para o manejo adequado. Neste trabalho, 26 populações do nematoide das galhas (Meloidogyne spp.), provenientes de municípios do estado da Bahia, Mato Grosso, Goiás e de Minas Gerais, foram caracterizados com base na morfologia da região perineal das fêmeas de Meloidogyne spp., seu perfil de esterase e identificação baseada em amplificação de regiões específicas do genoma dessas populações. Entre as populações de Meloidogyne spp. obtidas, identificou-se M. incognita e M. javanica. Não houve presença de populações mistas nas amostras analisadas. O diagnóstico baseado em análise molecular se mostrou mais rápido e confiável comparado ás outras análises.

Biocontrol agents in the management of Meloidogyne incognita in tomato / Agentes de biocontrole no manejo de Meloidogyne incognita em tomateiro

ABSTRACT: Recognized as prominent among the principal pests of tomato crop, the root-knot nematode (Meloidogyne incognita) has generated much study with regards to control methods. This study aimed to assess the efficiency of biological products in decreasing the M. incognita population in tomato. Four experiments were conducted under greenhouse conditions. Two of these experiments had the purpose to determine the efficiency of NemOut™ (Bacillus subtilis + B. licheniformis + Trichoderma longibrachiatum), and the other two to assess the efficiency of other different biological products in M. incognita management. Tomato plants of the cultivar Santa Cruz Kada Giant were inoculated using 2000 eggs and J2 of M. incognita. In the first two experiments doses corresponding to 0, 4, 6, 8 and 10kg ha-1 of NemOut™ were applied after inoculation. In the second study, the treatments consisted of the use of biological products containing different other biocontrol agents. Assessments were made at 45 and 65 days after inoculation (DAI) to evaluate plant height, fresh shoot and fresh root weigh, gall index and egg mass index, population density and reproduction factor (RF). The NemOut™ doses reduced the M. incognita population density and RF at 45 DAI. The agent Pochonia chlamydosporia showed greater efficiency in controlling M. incognita on tomato. Biological products and the different doses of NemOut™ had no influence on the tomato plant development.

RESUMO: O nematoide das galhas (Meloidogyne incognita) é relatado como uma das principais pragas da cultura do tomate, sendo importante o estudo de alternativas para o seu manejo. O presente estudo objetivou avaliar a eficiência de produtos biológicos na redução da população de M. incognita em tomate. Foram conduzidos quatro experimentos em casa de vegetação, dois visando avaliar a eficiência de doses de NemOut™ (Bacillus subtilis + B. licheniformis + Trichoderma longibrachiatum), e outros dois para avaliar a eficiência de diferentes produtos biológicos no controle de M. incognita. Plantas de tomate Santa Cruz Kada Gigante foram inoculadas com 2000 ovos e J2 de M. incognita. No primeiro estudo foram utilizadas doses equivalentes a 0, 4, 6, 8 e 10kg ha-1 de NemOut™, aplicadas após a inoculação. No segundo estudo os tratamentos constituíram de produtos biológicos com diferentes agentes de biocontrole. As avaliações foram realizadas 45 e 65 dias após a inoculação (DAI), determinando-se altura de plantas, massa fresca da parte aérea e da raiz, índice de galhas e de massas de ovos, densidade populacional e fator de reprodução (FR). Doses crescentes de NemOut™ reduziram a densidade populacional e o FR de M. incognita, aos 45 DAI. O agente Pochonia chlamydosporia mostrou-se mais eficiente no controle de M. incognita no tomateiro. Os produtos biológicos e as diferentes doses de NemOut™ não afetaram o desenvolvimento das plantas de tomate.

Disease of root-knot nematode and control strategy in medicinal plants / 中国中药杂志

Root-knot nematode (Meloidogyne spp.) is one of the important disease of medicinal plant cultivation and seriously hinders the sustainable development of traditional Chinese medicine industry. We introduced the main species, identification methods and control strategies of root-knot nematode diseases in the medicinal plants in this study. Identifications of morphology and molecular were the main tools for the distinction of root-knot nematodes at present. This study stated that integrated system was established for root-knot nematode control, including that integrated control technique was the first step, disease-resistant varieties with high yield were the basis, and normalized patterns of cultivation and management were the measure. These strategies would improve the sustainable development of medicinal plants and environmental protection.

Grafting guava on cattley guava resistant to Meloidogyne enterolobii / Enxertia da goiabeira sobre araçazeiros resistentes a Meloidogyne enterolobii

The use of resistant rootstocks could be a promising method to control nematodeMeloidogyne enterolobiiin commercial plantations of guava. The present study aimed to evaluate the success of grafting guava as a scion on accessions of cattley guava as rootstocks resistant to M. enterolobii.The treatments consisted of the rootstocks cattley guava plants (three accessions of Psidium cattleyanum) and common guava (control). In the apical wedge grafting method, scion of Paluma cultivated variety was used. The experiment was arranged in a randomized block design with four treatments and five replicates, and eight plants per plot. The saplings produced as described before were planted in the field where the initial growth of the different combinations were evaluated. Graft success was observed for the control (common guava) and for accessions 115 and 117 of cattley guava plants, with success rates of 63, 32 and 29%, respectively. In the field, the cattley guava used as rootstocks hampered Paluma canopy development and caused death of plants. Incompatibility of P. cattleyanumas rootstocks for P. guajavaPaluma was confirmed one year after cultivation in field.

O uso de porta-enxertos resistentes poderá ser um método promissor para o controle do nematoide Meloidogyne enterolobiiem plantios comerciais de goiaba, em caso de compatibilidade de enxertia. O presente trabalho teve como objetivo avaliar o pegamento via enxertia por garfagem de topo em fenda cheia, entre a goiabeira e acessos de araçazeiros resistentes a M. enterolobii. Os tratamentos foram constituídos pelos porta-enxertos utilizados, araçazeiros (três acessos de Psidium cattleyanum) e a goiabeira. O delineamento estatístico adotado foi em blocos casualizados (DBC), com quatro tratamentos, cinco repetições, e oito plantas por parcela. Após a enxertia, foi avaliada a percentagem de pegamento dos enxertos. As mudas produzidas foram plantadas no campo e avaliadas quanto ao crescimento inicial das diferentes combinações. Constatou-se pegamento de enxertia entre a goiabeira 'Paluma' e os porta-enxertos de goiabeira e araçazeiros dos acessos 115 e 117, ocorrendo uma taxa de pegamento de 63, 32 e 29%, respectivamente. No campo, os araçazeiros utilizados como porta-enxertos induziram menor desenvolvimento de copa e resultaram em morte de plantas. A incompatibilidade entre a goiabeira 'Paluma' e o porta-enxerto P. cattleyanumfoi confirmada um ano após plantio no campo.

Use of the arbuscular mycorrhizal fungus Glomus intraradices as biological control agent of the nematode Nacobbus aberrans parasitizing tomato

The plant-parasitic nematode Nacobbus aberrans is an endoparasite that induces gall formation in the roots and causes severe losses to diverse crops. Some populations of this nematode show preference for certain hosts, revealing the existence of "races/groups" with different behaviour and making nematode management difficult. A possible biological control alternative to reduce the damage caused by this species may be the use of arbuscular mycorrhizal fungi (AMF). In the present work, the effect of Glomus intraradices on tomato plants inoculated with the nematode at transplanting and three weeks later was tested. At 60 days, the following parameters were estimated: percentage of AMF colonization, root and aerial dry weight, number of galls and egg masses, and reproduction factor (RF=final population/initial population) of N. aberrans. AMF colonization was higher in the presence of the nematode. The use of AMF favoured tomato biomass and reduced the number of galls and RF on the plants inoculated with the nematode at transplanting.

Incorporação ao solo de substrato contendo micélio e conídios de Pochonia chlamydosporia para o manejo de Meloidogyne javanica / Soil amendment with substrate containing mycelium and conidia of Pochonia chlamydosporia for the management of Meloidogyne javanica

Os clamidósporos são os principais propágulos utilizados como fonte de inóculo de Pochonia chlamydosporia em experimentos envolvendo o biocontrole do nematoide das galhas. O presente trabalho teve por objetivo avaliar o controle de Meloidogyne javanica em tomateiro por meio da aplicação ao solo de grãos de arroz colonizados pelo fungo contendo apenas micélio e conídio, sem a presença de clamidósporos. O isolado de P. chlamydosporia Pc-10 foi cultivado por 15 dias a 26°C em arroz previamente esterilizado em forno microondas. Dois experimentos foram conduzidos simultaneamente em casa de vegetação. No experimento 1, vasos de 2L de capacidade foram preenchidos com mistura solo:areia (1:1, v:v), contendo 3g kg-1 de solo de grãos de arroz colonizados pelo antagonista. No experimento 2, o fungo foi adicionado ao solo de vasos de 0,5L nas doses de 1, 5, 10, 15, 20, 25 ou 30g kg-1 de solo. Em seguida, o substrato de cada vaso foi infestado com 4.000 ovos de M. javanica e, após 15 dias, uma plântula de tomate foi transplantada. No experimento 1, a aplicação do fungo ao solo reduziu o número de galhas e de ovos do nematoide em 40% e 72,83%, respectivamente. No experimento 2, houve redução do número de ovos a partir de doses de 5g kg 1 de solo e no número de galhas, principalmente, nas doses de 25 e 30g kg-1 de solo. Conclui-se que P. chlamydosporia Pc-10 controlou M. javanica em tomateiro, mesmo quando aplicado ao solo na forma de grãos de arroz colonizados e sem a presença de clamidósporos.

Chlamydospores are the main propagules used as source of inoculum of Pochonia chlamydosporia in biocontrol experiments of root-knot nematodes. The objective of this study was to evaluate the control of Meloidogyne javanica on tomato plants by the soil application of rice grains colonized by Pochonia chlamydosporia containing just mycelium and conidia, without chlamydospores. The fungus (isolate Pc-10) was grown for 15 days at 26°C on grains of rice, previously sterilized in microwave oven. Two experiments were simultaneously carried out under greenhouse conditions. In the experiment 1, 2-L pots were filled with a soil:sand mixture (1:1, v:v) containing 3g kg-1 of soil of rice grains colonized by the antagonist. In the experiment 2, the fungus was added into the soil of 0.5L pots at the doses of 5, 10, 15, 20, 25 or 30g kg-1 of soil. The soil of each pot was infested with 4,000 eggs of M. javanica and one tomato seedling was transplanted in each pot after fifteen days. In the experiment 1, the application of the fungus into the soil reduced the number of galls and eggs of the nematode by 40% and 72.83%, respectively. In the experiment 2, it was observed the reduction of the number of eggs from the dose of 5 g kg-1 of soil and of the number of galls, particularly at the doses of 25 and 30g kg-1 of soil. As a conclusion, P. chlamydosporia controlled M. javanica on tomato plants even when applied into the soil as colonized-rice grains and without chlamydospores.

Essential oil of Rosmarinus officinalis in the control of Meloidogyne javanica and Pratylenchus brachyurus in soybean / ÓLeo essencial de Rosmarinus officinalis no controle de Meloidogyne javanica E Pratylenchus brachyurus em soja

The aim of this study was to assess the nematicidal activity of the essential oil of Rosmarinus officinalis applied directly to the soil to control Meloidogyne javanica and Pratylenchus brachyurus in soybean. The experiments were conducted in the greenhouse in a fully randomized design with a 4 x 2 factorial arrangement consisting of four concentrations of oil (0, 1, 2 and 3%) and two treatment frequencies (fortnightly and monthly), with four replications. Plantlets of soybean cv. CD 206 were transplanted to 1-liter pots and the first treatment applied one day after transplanting. One week after this treatment, the pots were inoculated with 2000 eggs of M. javanica or 1000 specimens of P. brachyurus per plant. The plants were collected 60 days after inoculation and plant growth variables assessed (height, aerial part fresh weight, root fresh weight and aerial part dry weight). The galls number and eggs per gram of root was assessed for M. javanica and the number of nematodes per gram of root and per 100 cm3 soil for P. brachyurus. Was observed that the population of M. javanica was reduced by R. officinalis essential oil, but none of the concentrations studied affected the population of P. brachyurus.

Objetivou-se avaliar a atividade nematicida do óleo essencial de Rosmarinus officinalis, aplicado diretamente ao solo, visando o controle de Meloidogyne javanica e Pratylenchus brachyurus em soja. Os ensaios foram conduzidos em casa de vegetação, em delineamento inteiramente casualizado, em esquema fatorial 4 x 2, sendo quatro concentrações do óleo (0, 1, 2 e 3%) e duas frequências de tratamento (quinzenal e mensal), com quatro repetições. Plântulas de soja cv. CD 206 foram transplantadas para vasos de 1 L, recebendo o primeiro tratamento um dia após o transplantio e inoculadas uma semana após esse tratamento com 2000 ovos de M. javanica ou 1000 espécimes de P. brachyurus por planta. As plantas foram coletadas 60 dias após a inoculação, avaliando-se variáveis vegetativas da cultura (altura, massa fresca de parte aérea e de raiz e massa seca de parte aérea); número de galhas e ovos por grama de raiz para M. javanica e nematoides por grama de raiz e por 100 cm3 de solo para P. brachyurus. Observou-se que a população de M. javanica foi reduzida pelo óleo essencial de R. officinalis, porém nenhuma das concentrações estudadas afetou a população de P. brachyurus.