Elizabeth de Oliveira

Growth and Nutrition of Mollicute-Infected Maize

Maize bushy stunt phytoplasma (MBSP) and corn stunt spiroplasma (CSS) diseases are widespread in Brazil. The leafhopper Dalbulus maidis is the insect vector for these pathogenic mollicutes. The effects of these diseases on the development of maize plants and the possible interaction of soil water availability on these effects were evaluated in two experiments carried out on potted plants. Experiment 1 was carried out in a 2 × 4 factorial, where factor 1 corresponded to healthy and mollicute-infected plants and factor 2 to the maintenance of 40, 60, 80, and 100% of the total soil water availability. Leafhoppers collected from a field with high incidence of these diseases were used to inoculate plants with mollicutes. There were three treatments in experiment 2: healthy plants, plants infected with phytoplasma, and plants infected with spiroplasma. MBSP was predominant in experiment 1. The infected plants grew less and lowered nutrient uptake, in distinct proportions, indicating a differential effect of mollicutes on nutrient uptake independent of available soil water. Soil water availability did not significantly affect plant growth and nutrient uptake or mollicute infection. The results indicated that plants infected by mollicutes contained less protein than healthy plants. Experiment 2 showed a reduction in growth of plants infected with mollicutes and less nutrient uptake by spiroplasma-infected plants. The results showed a detrimental effect of the spiroplasma on Mg uptake. Both experiments showed more water retention by infected plants than by healthy ones. These experiments clearly demonstrated that reduced plant growth and nutrient uptake are major effects on plants infected with MBSP and CSS.

Occurrence of Maize rayado fino virus in Maize in Argentina

Symptoms of fine chlorotic stipple-striping of the veins, chlorosis, numerous dots and stripes, formation of holes in the leaf blade, and ears reduced in size, bearing few grains, were observed in maize crops in Tafí del Valle (Tucumán Province), Orán, El Galpón (Salta Province), Tilcara and Yaví (Jujuy Province), the subtropical area of northwest Argentina where the leafhopper vector Dalbulus maidis (DeLong & Wolcott) is present. Maize rayado fino virus (MRFV) was detected in these samples by a positive reaction in double antibody sandwich-enzyme-linked immunosorbent assay (DAS-ELISA) using an AGDIA kit. Electron microscopy revealed abundant isometric particles about 30 nm in diameter in the cytoplasm and vacuoles of phloem cells and xylem parenchyma cells. The virus was also detected by reverse transcription polymerase chain reaction (RT-PCR) using a primer pair MRFV-09/MRFV-10. Primers and PCR conditions were as previously described (1). Virus amplification was observed only in samples from symptomatic plants. In 1981 (2), the presence of MRFV in Argentina was revealed by serological assay in plants from temperate central areas. No further reports were released since then. This is the first evidence of MRFV in subtropical areas of Argentina and identification of the virus by combining DAS-ELISA, particle size, relation with plant tissues, and RTPCR. References: (1) R. W. Hammond et al. J. Gen. Virol. 78:3153, 1997. (2) S. F. Nome et al. RIA XIX:257, 1984.

Genetic variability of Brazilian phytoplasma and spiroplasma isolated from maize plants

The objective of this work was to characterize the genetic variability of phytoplasma and Spiroplasma kunkelii isolated from maize plants showing symptoms of stunt collected from different Brazilian geographic regions. A DNA fragment of 500 base pairs (bp) was amplified from the spiralin gene in S. kunkelii and one fragment of 1,200 bp was generated from 16S rDNA gene in phytoplasma. The partial sequences of the spiralin gene showed similarity of 98% among the isolates of S. kunkelii analyzed. These sequences were compared with the sequence of the spiralin gene from other Spiroplasma species deposited in the GenBank, resulting in a similarity varying from 76.9% to 88.1%. The 16S rDNA sequence from the phytoplasma were completely similar within the Brazilian isolates and showed up to 98% of the similarity with sequences already found from other phytoplasmas. A very narrow genetic variability was detected by these gene fragments within phytoplasma and Spiroplasma analyzed. However, other genomic regions with higher polymorphic levels shall be identified in order to better evaluate the genetic diversity within these microorganisms population.

ABUNDANCE AND SPECIES RICHNESS OF LEAFHOPPERS AND PLANTHOPPERS (HEMIPTERA: CICADELLIDAE AND DELPHACIDAE) IN BRAZILIAN MAIZE CROPS

Abstract Insects in the Cicadellidae and Delphacidae families, common in grasses, are an important group of vectors of viruses and mollicutes, which cause diseases in several plant species. The goal of this study was to evaluate the abundance and species richness of Cicadellidae and Delphacidae and the presence of potential vectors of viruses and mollicutes in maize crops in Brazil. Insects were collected using sweep nets in maize crops in 48 counties of 8 states, distributed in 4 regions of Brazil in the yr 2005, 2006 and 2007, with a total of 198 samples. The collected material was screened, and the leafhoppers and planthoppers were identified at the species level. A total of 4233 Cicadellidae specimens, including 30 species, and 205 Delphacidae specimens, including 9 species, were collected. The most abundant species was Dalbulus maidis (Delong & Wolcott) representing 90.1% of leafhoppers and planthoppers. Abundance and species richness differences were observed between the northeast and center-south regions of Brazil. Eight species of leafhoppers and planthoppers were identified as vectors or potential vectors of viruses and mollicutes in maize, although some of these viruses are not reported in Brazil. Among these species, we identified the planthoppers, Caenodelphax teapae (Fowler), Peregrinus maidis Ashmed, Pyrophagus tigrinus Remes Lenicov & Varela and Toya propinqua (Fieber) are experimental vectors of Mal de Río Cuarto virus, which is a quarantine virus in Brazil. Two Cicadellidae species and 3 Delphacidae species were reported for the first time in Brazil.

Simultaneous transmission of phytoplasma and spiroplasma by Dalbulus maidis leafhopper and symptoms of infected maize

Two experiments were carried out to study simultaneous transmission of phytoplasmas and spiroplasmas by the leafhopper Dalbulus maidis from double infected popcorn-plants. The double infection was obtained through only one leafhopper from field per plant. The results indicated variability among isolates and combinations of phytoplasmas and spiroplasmas for transmission by D. maidis and maize leaf symptoms expression.

Abundance of the insect vector of two different Mollicutes plant pathogens in the vegetative maize cycle

The abundance of Dalbulus maidis leafhopper was evaluated in intervals of 20 days in the vegetative cycle of maize and popcorn hybrids simultaneously cultivated in three areas at four different sowing dates. The amount of leafhoppers increased from the first until the third sampling, around the flowering, and decreased after that. The highest amount of leafhoppers detected in the late sowings was attributed to effect of their concentration derived from oldest maize crops.

Fatores relacionados à incidência e disseminação do vírus do mosaico comum do milho

In recent years, mosaic became one of the most important diseases affecting maize (Zea mays) crops. It is caused by a potyvirus complex transmitted by aphid species. The aim of this work was to identify factors that may contribute to increase the incidence and dissemination of this disease. Seedlings of 115 cultivars from three different National Maize Cultivars Trials were tested for their susceptibility to the virus complex via mechanical inoculation of five plants of each cultivar, between October/97 and February/98. Most cultivars were susceptible and showed mosaic symptoms 15 days after inoculation. Several graminaceous species were also inoculated and showed to be hosts of the potyvirus complex. Using the dot-ELISA test, the virus complex could be detected in maize plants collected from different regions of Sao Paulo, Minas Gerais and Goias states. Between March/97 and February/98 two maize inbred lines were planted monthly and incidence of mosaic was evaluated weekly. The highest incidence was observed in the summer, coinciding with the increase of temperature and the levels of rainfall. These results may contribute to recommending control measures.

Occurrence and Epidemiological Aspects of Phytoplasmas in Cereals

The symptoms of the main cereal diseases associated with phytoplasma presence such as barley deformation, maize bushy stunt, maize redness, green ear, oat proliferation, oat stunt, oat yellows, rice orange leaf, rice yellow dwarf, triticosecale stunt, wheat blue dwarf and wheat streak, yellowing, and stunting are described. Moreover the phytoplasmas associated with each disease are described together with their geographical distribution. In some cases, factors that affect the occurrence, development, and severity of these diseases are discussed. The insect vectors involved in transmission of some of these diseases are presented, along with the main morphological characteristics adopted for their identification.

Genetic control of maize resistance to corn stunt spiroplasma

The corn stunt spiroplasma (CSS) is transmitted by Dalbulus maidis leafhopper. The disease causes severe losses on maize production. One complete diallel genotype of two corn stunt resistant maize lines, and two corn stunt susceptible maize lines were tested by spiroplasma inoculation under insect-proof screenhouse condition, in order to elucidate the maize genetic control to this disease. The percentage of reduction on plant dry weight caused by the spiroplasma inoculation in relation to the dry weight average of the healthy plants, was calculated for each plant spiroplasma inoculated, to express maize resistance. The disease incidence for each treatment was represented by the percentage of plants with CSS symptoms. The disease severity was scored based on CSS symptoms severity, for each plant. Reductions on the dry weight were caused by the disease; the disease severity data showed a positive and strong correlation (r = 0.80). The statistical analyses indicated that genetic control of maize resistance to spiroplasma includes genes with additive effects and genes with dominance or epistatic effects, suggesting complex resistance inheritance. One line was better performing than all the others, reducing the disease damage, as well per se as in crosses.

ENFEZAMENTOS EM MILHO: EXPRESSÃO DE SINTOMAS FOLIARES,

Com o objetivo de verificar a multiplicacao, expressao de sintomas foliares e efeito dos molicutes na producao de diferentes genotipos de milho, fitoplasma e Spiroplasma kunkelii foram inoculados, isolada ou simultaneamente, em cinco cultivares, mantidas em vasos ate a producao. Plantas sadias de todas as cultivares foram utilizadas como controle. Em cada vaso, foram cultivadas duas plantas, sendo uma utilizada para a deteccao dos molicutes na ultima folha completamente expandida, aos 30, 60 e 100 dias apos a inoculacao. A outra planta foi utilizada para deteccao dos molicutes nas folhas inferiores, medianas e apicais, aos 100 dias apos a inoculacao. O percentual de reducao causado pelos molicutes na altura e na producao de graos foi determinado nas duas plantas, em relacao as plantas sadias. A expressao de sintomas foliares e os resultados positivos obtidos na deteccao dos molicutes foram mais evidentes aos 100 dias apos a inoculacao. O periodo de enchimento de graos foi considerado a fase mais adequada para a deteccao desses patogenos. Os molicutes foram detectados nas folhas inferiores, medianas e apicais, observando-se maior frequencia nas folhas apicais. Verificou-se predominância de infeccao por espiroplasma e maior efeito detrimental desse patogeno no crescimento e producao das plantas em relacao ao fitoplasma. Nao foi detectado efeito sinergistico significativo dos dois molicutes sobre o crescimento e producao das plantas. Observou-se que o efeito prejudicial desses patogenos sobre o crescimento e producao das cultivares foi determinado pela frequencia de plantas infectadas.Apesar do amplo uso de deltametrina para controlar pragas do milho, no Brasil, existem poucas informacoes sobre o seu impacto nos artropodes associados a cultura. Este trabalho teve como objetivo avaliar os efeitos da pulverizacao de deltametrina em atropodes pragas e predadores mais comuns na parte aerea do milho. O delineamento foi em blocos casualizados, com quatro repeticoes. No estadio de dez folhas expandidas, o inseticida (Decis 25 CE, 7,5 gi.a./ha) foi pulverizado no cartucho das plantas e amostragens realizadas em dez plantas/parcela (1 dia antes, 7, 10, 14, 21 e 28 dias apos a pulverizacao) determinaram a densidade populacional ou a ocorrencia dos artropodes e a percentagem de plantas atacadas por Spodoptera frugiperda . O inseticida controlou a infestacao de S. frugiperda ate sete dias apos a pulverizacao e, nesse periodo, tambem reduziu em 64% a densidade populacional da cigarrinha Dalbulus maidis nas parcelas tratadas. O inseticida nao afetou negativamente o complexo de artropodes predadores associados a parte aerea do milho, mostrando toxicidade seletiva em favor de ninfas e adultos de Doru luteipes , de um grupo nao identificado de formigas e de aranhas. Esses resultados sao favoraveis ao uso de deltametrina em programas de manejo de S. frugiperda na cultura do milho, principalmente nos agroecossistemas com elevada abundância de predadores.Com o objetivo de estudar o efeito da omissao de P no meio de cultura nas caracteristicas fisiologicas de absorcao de P e no comprimento de raizes de oito genotipos de milho, quatro previamente classificados como eficientes e quatro como ineficientes para P, foram desenvolvidos dois experimentos. No primeiro, sementes dos oito hibridos de milho foram germinadas em rolos de papel-toalha e transplantadas para recipientes de plastico contendo solucao nutritiva completa de Steinberg, pH 5,5. Sete dias apos o transplante, o P foi retirado de metade das bandejas. Apos tres dias de omissao de P, realizou-se a cinetica de absorcao de P, iniciando-se a retirada de aliquotas de 10 ml de cada vaso, a cada 30 minutos, durante oito horas. No segundo experimento, aos seis dias apos a omissao do P no meio de cultivo, raizes das plantas dos tratamentos com e sem omissao de fosforo foram coletadas para avaliacao do comprimento total do sistema radicular. A omissao de fosforo por tres dias aumentou significativamente a velocidade maxima de absorcao de fosforo (Vmax) e a taxa de influxo em seis genotipos de milho, quatro ineficientes e dois eficientes para P. O Km (Constante de Michaelis) foi significativamente modificado pela omissao de fosforo em apenas dois genotipos. Conclui-se que aumentos na Vmax e taxa de influxo de P com a omissao de P no meio de cultivo e um mecanismo geral de resposta das plantas a deficiencia de P, mas que nao confere eficiencia diferencial entre genotipos de milho. A omissao de P na solucao nutritiva, por seis dias, aumentou significativamente o comprimento radicular em tres dos genotipos eficientes para P. Nos genotipos ineficientes para P, nao houve modificacao significativa dessas caracteristicas, indicando que o crescimento diferencial do sistema radicular pode ser um dos possiveis mecanismos da eficiencia para fosforo em milho.