Yield loss caused by huanglongbing in different sweet orange cultivars in So Paulo, Brazil

Citrus huanglongbing (HLB) reduces an affected orchard`s economic life. This work aimed to characterize yield loss due to HLB for different sweet orange cultivars and determine the relationship between disease severity and yield. Disease severity and yield were assessed on 949 individual trees distributed in 11 different blocks from sweet orange cultivars Hamlin, Westin, Pera and Valencia. In each block, plants showing a range of HLB severity levels and asymptomatic plants were selected. Total yield (weight of harvested fruit), mean weight of asymptomatic and symptomatic fruit, relative yield (symptomatic tree yield/mean yield of asymptomatic trees from the same block) and relative number of fruits (fruit number from symptomatic tree/mean number of fruits from asymptomatic trees from the same block) were determined. The weight of symptomatic fruit was lower than the weight of asymptomatic fruit, but the weights of asymptomatic and symptomatic fruit were not correlated with disease severity, indicating that the effects of HLB were restricted to symptomatic branches. The relationship of the relative yield with HLB severity can be satisfactorily described by a negative exponential model. The rates of yield decrease as a function of disease severity were similar for all assessed cultivars. A relative yield (up to 19%) was observed even for trees where disease severity was 100%. The strong linear relationship between relative number of fruits per tree and the relative yield per tree suggested that the yield reduction was due primarily to early fruit drop or lack of fruit set on affected branches.

Phytoplasma closely related to the pigeon pea witches`-broom phytoplasma (16Sr IX) is associated with citrus huanglongbing symptoms in the state of Sao Paulo, Brazil

In February 2007, sweet orange trees with characteristic symptoms of huanglongbing (HLB) were encountered in a region of Sao Paulo state (SPs) hitherto free of HLB. These trees tested negative for the three liberibacter species associated with HLB. A polymerase chain reaction (PCR) product from symptomatic fruit columella DNA amplifications with universal primers fDI/rPI was cloned and sequenced. The corresponding agent was found to have highest 16S rDNA sequence identity (99%) with the Pigeon pea witches`-broom phytoplasma of group 16Sr IX. Sequences of PCR products obtained with phytoplasma 16S rDNA primer pairs fU5/rU3, fU5/P7 confirm these result.,;. With two primers D7f2/D7r2 designed based oil the 16S rDNA Sequence of the cloned DNA fragment, positive amplifications were obtained from more than one hundred samples including symptomatic fruits and blotchy mottle leaves. Samples positive for phytoplasmas were negative for liberibacters, except for four samples, which were positive for both the phytoplasma and `Candidatus Liberibacter asiaticus`. The phytoplasma was detected by electron microscopy in the sieve tubes of midribs from symptomatic leaves. These results Show that a phytoplasma of group IX is associated with citrus HLB symptoms ill northern, central, and Southern SPs. This phytoplasma has very probably been transmitted to citrus from an external Source of inoculum, but the Putative insect vector is not yet known.

Distribuição espacial de Huanglongbing (Greening) em citros utilizando a geoestatística

Os objetivos do trabalho foram avaliar a distribuição espacial e a expansão da Huanglongbing (greening) em talhões de citros de uma propriedade agrícola localizada no município de Araraquara-SP, utilizando a geoestatística. Para determinar o número de plantas com greening, foram realizadas inspeções periódicas em intervalos de três meses, no período de março de 2005 a julho de 2007, contando-se, em cada talhão, o número de plantas com os sintomas característicos da doença. Realizou-se a análise descritiva dos dados e, para verificar a distribuição espacial do greening, utilizou-se a geoestatística através do ajuste de semivariogramas e da interpolação dos dados por krigagem. A dependência espacial de plantas com greening apresentou raio de agregação de 300 a 560 m, indicando distribuição agregada da doença. Por meio dos mapas de krigagem, observou-se que o foco inicial de plantas doentes ocorreu nos limites da fazenda, com expansão do greening por toda a área. O intervalo de inspeção de três meses não foi adequado para a redução do greening na fazenda.

Estimativa dos impactos econômicos decorrentes de eventual introdução do huanglongbing (HLB) no estado da Bahia

A Bahia é o segundo mais importante Estado produtor de citros do Brasil, responsável por 5,5% da produção do País. 80% da produção vem de propriedades de base familiar, as quais dependem desse cultivo para seu sustento econômico. O Huanglongbing (HLB) nunca foi registrado na Bahia, mas está disseminando-se em outros três Estados do Brasil (São Paulo, Paraná e Minas Gerais), um dos quais faz fronteira com a Bahia. Assim, este estudo teve como objetivo estimar o impacto econômico potencial de uma eventual introdução do HLB na Bahia. Foram utilizados os modelos de Gompertz e logístico associados a diferentes estratos de idade das plantas para estimar, respectivamente, a evolução da incidência e da severidade da doença no parque citrícola baiano, num horizonte de 20 anos. O progresso da doença foi usado para estimar os danos à produção em três cenários. No cenário-base (A), os esforços da agência estadual de defesa agropecuária previniria a entrada do HLB. No cenário B, a doença seria introduzida e não haveria esforços para seu controle, levando a uma disseminação nos anos seguintes. No cenário C, após a detecção da doença, os produtores adotariam medidas de controle: eliminação de plantas sintomáticas e supressão das populações do inseto vetor. Os custos do controle foram calculados considerando-se a necessidade de pulverizações, inspeções periódicas e eliminação das plantas doentes. O valor presente líquido (VPL) foi usado para comparar os três cenários. Os resultados indicam que as perdas seriam muito significativas caso o HLB fosse introduzido na Bahia. Assim sendo, se as medidas de controle não fossem aplicadas, os prejuízos poderiam superar R$1,8 bilhão.

Detección de los agentes causales de la enfermedad huanglongbing de los cítricos, candidatus liber.

Tesis (Maestría en Ciencias en Producción Agrícola) UANL, 2012.

Estrategias biotecnológicas para el control de diaphorina citri vectror de la bacteria candidatus liberibacter asiaticus, agente causal de huanglongbing.

Tesis (Doctor en Ciencias con acentuación en Química de Productos Naturales) UANL, 2014.

Distribuição espacial de Huanglongbing (Greening) em citros utilizando a geoestatística

Os objetivos do trabalho foram avaliar a distribuição espacial e a expansão da Huanglongbing (greening) em talhões de citros de uma propriedade agrícola localizada no município de Araraquara-SP, utilizando a geoestatística. Para determinar o número de plantas com greening, foram realizadas inspeções periódicas em intervalos de três meses, no período de março de 2005 a julho de 2007, contando-se, em cada talhão, o número de plantas com os sintomas característicos da doença. Realizou-se a análise descritiva dos dados e, para verificar a distribuição espacial do greening, utilizou-se a geoestatística através do ajuste de semivariogramas e da interpolação dos dados por krigagem. A dependência espacial de plantas com greening apresentou raio de agregação de 300 a 560 m, indicando distribuição agregada da doença. Por meio dos mapas de krigagem, observou-se que o foco inicial de plantas doentes ocorreu nos limites da fazenda, com expansão do greening por toda a área. O intervalo de inspeção de três meses não foi adequado para a redução do greening na fazenda.

Ineffectiveness of pruning to control citrus huanglongbing caused by Candidatus Liberibacter americanus

The huanglongbing (HLB) disease of citrus trees, caused by Candidatus Liberibacter asiaticus and Ca. Liberibacter americanus, was first reported in Brazil in March, 2004. The presence of the disease has caused serious concerns among growers. Pruning experiments were conducted to determine if removal of symptomatic branches or the entire canopy (decapitation) would eliminate infected tissues and save HLB-affected trees. Pruning was done in five blocks on a total of 592 3- to 16 year-old 'Valencia', 'Hamlin' or 'Pera' sweet orange trees showing no symptoms or with two levels of symptom severity. Ten decapitated trees per block were caged and all trees were treated with insecticides to control the psyllid vector, Diaphorina citri. Mottled leaves reappeared on most symptomatic (69.2%) as well on some asymptomatic (7.6%) pruned trees, regardless of age, variety, and pruning procedure. Presence of the pathogen (Ca. Liberibacter americanus) in all symptomatic trees was confirmed by PCR. In general, the greater the symptom severity before pruning the lower the percentage of trees that remained asymptomatic after pruning.

Candidatus Liberibacter americanus, associated with citrus huanglongbing (greening disease) in São Paulo State, Brazil

Symptoms of huanglongbing (HLB) were reported in São Paulo State (SPS), Brazil, in March 2004. In Asia, HLB is caused by 'Candidatus Liberibacterasiaticus'and in Africa by 'Candidatus Liberibacter africanus'. Detection of the liberibacters is based on PCR amplification of their 16S rRNA gene with specific primers. Leaves with blotchy mottle symptoms characteristic of HLB were sampled in several farms of SIPS and tested for the presence of liberibacters. 'Ca. L. asiaticus' was detected in a small number of samples but most samples gave negative PCR results. Therefore, a new HLB pathogen was suspected. Evidence for an SPS-HLB bacterium in symptomatic leaves was obtained by PCR amplification with universal primers for prokaryotic 16S rRNA gene sequences. The amplified 16S rRNA gene was cloned and sequenced. Sequence analysis and phylogeny studies showed that the 16S rRNA gene possessed the oligonucleotide signatures and the secondary loop structure characteristic of the alpha-Proteobacteria, including the liberibacters. The 16S rRNA gene sequence phylogenetic tree showed that the SPS-HLB bacterium clustered within the a-Proteobacteria, the liberibacters being its closest relatives. For these reasons, the SPS-HLB bacterium is considered a member of the genus 'Ca. Liberibacter'. However, while the 16S rRNA gene sequences of 'Ca. L. asiaticus' and 'Ca. L. africanus' had 98-4% similarity, the 16S rRNA gene sequence of the SPS-HLB liberibacter had only 96(.)0% similarity with the 16S rRNA gene sequences of 'Ca. L. asiaticus'or'Ca. L. africanus'. This lower similarity was reflected in the phylogenetic tree, where the SPS-HLB liberibacter did not cluster within the 'Ca. L asiaticus'/'Ca. L. africanus group', but as a separate branch. Within the genus 'Candidatus Liberibacter' and for a given species, the 16S/23S intergenic region does not vary greatly. The intergenic regions of three strains of 'Ca. L. asiaticus', from India, the People's Republic of China and Japan, were found to have identical or almost identical sequences. In contrast, the intergenic regions of the SPS-HLB liberibacter, 'Ca. L. asiaticus' and 'Ca. L. africanus' had quite different sequences, with similarity between 66(.)0 and 79(.)5%. These results confirm that the SPS-HLB liberibacter is a novel species for which the name 'Candidatus Liberibacter americanus' is proposed. Like the African and the Asian liberibacters, the 'American' liberibacter is restricted to the sieve tubes of the citrus host. The liberibacter could also be detected by PCR amplification of the 16S rRNA gene in Diaphorina citri, the psyllid vector of 'Ca. L. asiaticus', suggesting that this psyllid is also a vector of 'Ca. L. americanus' in SPS. 'Ca. L. americanus' was detected in 216 of 218 symptomatic leaf samples from 47 farms in 35 municipalities, while 'Ca. L. asiaticus' was detected in only 4 of the 218 samples, indicating that 'Ca. L. americanus' is the major cause of HLB in SIPS.

Flutuação populacional e distribuição espacial de Diaphorina citri Kuwayama (Hemiptera: Liviidae) em propriedades citrícolas com manejo intensivo do Huanglongbing

Pós-graduação em Agronomia (Entomologia Agrícola) - FCAV

Estatística espacial e redes neurais aplicadas no estudo de epidemias de huanglongbing e mancha preta na cultura dos citros

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Parâmetros fisiológicos de laranjeiras com Huanglongbing

Pós-graduação em Agronomia (Produção Vegetal) - FCAV

Weedy Hosts and Prevalence of Potential Leafhopper Vectors (Hemiptera: Cicadellidae) of a Phytoplasma (16SrIX group) Associated With Huanglongbing Symptoms in Citrus Groves

Huanglongbing (HLB) is a severe citrus (Citrus spp.) disease associated with the bacteria genus Candidatus Liberibacter, detected in Brazil in 2004. Another bacterium was found in association with HLB symptoms and characterized as a phytoplasma belonging to the 16SrIX group. The objectives of this study were to identify potential leafhopper vectors of the HLB-associated phytoplasma and their host plants. Leafhoppers were sampled every other week for 12 mo with sticky yellow cards placed at two heights (0.3 and 1.5 m) in the citrus tree canopy and by using a sweep net in the ground vegetation of two sweet orange, Citrus sinensis (L.) Osbeck, groves infected by the HLB-phytoplasma in Sao Paulo state. Faunistic analyses indicated one Agalliinae (Agallia albidula Uhler) and three Deltocephalinae [Balclutha hebe (Kirkaldy), Planicephalus flavicosta (Stal), and Scaphytopius (Convelinus) marginelineatus (Stal)] species, as the most abundant and frequent leafhoppers (Hemiptera: Cicadellidae). Visual observations indicated an association of leafhopper species with some weeds and the influence of weed species composition on leafhopper abundance in low-lying vegetation. S. marginelineatus and P. flavicosta were more frequent on Sida rhombifolia L. and Althernantera tenella Colla, respectively, whereas A. albidula was observed more often on Conyza bonariensis (L.) Cronq. and B. hebe only occurred on grasses. DNA samples of field-collected S. marginelineatus were positive by polymerase chain reaction and sequencing tests for the presence of the HLB-phytoplasma group, indicating it as a potential vector. The association of leafhoppers with their hosts may be used in deciding which management strategies to adopt against weeds and diseases in citrus orchards.