Papaya Lethal Yellowing Virus Research Articles

Screening of papaya accessions resistant to Papaya lethal yellowing virus and capacity of Tetranychus urticae to transmit the virus

The objective of this work was to produce a polyclonal antiserum against the coat protein (CP) of Papaya lethal yellowing virus (PLYV) and to determine its specificity and sensibility in the diagnosis of the virus, as well as to evaluate the genetic resistance to PLYV in papaya (Carica papaya) accessions and to investigate the capacity of the two-spotted spider mite Tetranychus urticae to acquire and transmit PLYV to the plants. Sixty-five papaya accessions were evaluated. For each accession, ten plants were mechanically inoculated using PLYV-infected plant extracts, and three plants were mock inoculated with phosphate buffer alone and used as negative controls. Ninety days after inoculation, newly-emerging systemic leaves were collected from the inoculated plants, and viral infection was diagnosed by indirect Elisa, using polyclonal antiserum sensible to the in vitro-expressed PLYV CP. Viral transmission by T. urticae was evaluated in greenhouse. The experiments were repeated twice. Polyclonal antiserum recognized the recombinant PLYV CP specifically and discriminated PLYV infection from infections caused by other plant viruses. Out of the 65 papaya accessions evaluated, 15 were considered resistant, 18 moderately resistant, and 32 susceptible. The two-spotted spider mite T. urticae was capable of acquiring PLYV, but not of transmitting it to papaya.

Analysis of the full-length genome sequence of papaya lethal yellowing virus (PLYV), determined by deep sequencing, confirms its classification in the genus Sobemovirus

Papaya lethal yellowing virus (PLYV) causes an economically important disease in papayas in northeastern Brazil. Based on biological and molecular properties, PLYV has been tentatively assigned to the genus Sobemovirus. We report the sequence of the full-length genome of a PLYV isolate from Brazil, determined by deep sequencing. The PLYV genome is 4,145 nt long and contains four ORFs, with an arrangement identical to that of sobemoviruses. The polyprotein and CP display significant sequence identity with the corresponding proteins of other sobemoviruses. Pairwise comparisons and phylogenetic analysis based on complete nucleotide sequences confirm the classification of PLYV in the genus Sobemovirus.

Genetic variability of papaya lethal yellowing virus isolates from Ceará and Rio Grande do Norte States, Brazil

The papaya (Carica papaya) is a fruit crop of great economic importance throughout the Brazilian northeast, which is responsible for 60% of the national output. Papayas in the states of Ceara and Rio Grande do Norte are affected by lethal yellowing disease, caused by papaya lethal yellowing virus (PLYV). Previous work suggested that PLYV is a putative sobemovirus. To assess the genetic variability of PLYV, foliar samples were collected in October 2008 and October 2009 in commercial fields from Ceara and Rio Grande do Norte states, and total RNA was extracted. Specific primers based on the sequence of a previously characterized PLYV isolate were used for the RT-PCR-based amplification of a 900 bp fragment corresponding to the central region of the viral genome. Fragments from 21 viral isolates were cloned and sequenced. Sequence analyses indicated >97% nucleotide sequence identity among the isolates, 94-100% identity with the previously sequenced PLYV isolate, and a lower but significant identity with sobemoviruses (43-48.5%). These results suggest a low genetic variability among PLYV isolates, and are in agreement with the provisional placement of PLYV in the genus Sobemovirus. Definitive taxonomic conclusions, however, can only be drawn after the determination of the full-length genomic sequence.

Papaya lethal yellowing virus

Papaya lethal yellowing virus

Biological, Physical, and Molecular Properties of a Papaya lethal yellowing virus Isolate.

Papaya (Carica papaya) is an important tropical fruit crop in northeastern Brazil. Lethal yellowing is a disease caused by Papaya lethal yellowing virus (PLYV) that occurs only in northeastern Brazil. The symptoms are characterized by progressive leaf yellowing and greenish circular spots on the fruits. The virus has isometric particles (30 nm), genomic single-stranded (ss)RNA (1.6 × 106 Da), and a coat protein with a single component (35 kDa). Although no biological vector has been confirmed, the virus is spreading every year. In the present study, an isolate of the virus was characterized biologically, serologically, physically, and molecularly, and its survival in dried tissue was evaluated. Approximately 309.5 mg of virus was purified per kilogram of infected papaya leaves, and a high-titer polyclonal antiserum (1:1,024,000) was obtained from an immunized rabbit. The virus was detected in dried roots and leaves maintained up to 120 days. Its physical properties were thermal inactivation point (80°C), longevity in vitro (60 days), and dilution end point (10-6). The virus was inactivated in leaves and roots eradicated from infected plants when they were submitted to a solarization period of 12 days, but maintained its infectivity when leaves and roots were maintained over the soil at natural conditions for 32 days. Among 58 plant species from 13 botanical families, the PLYV isolate infected only C. papaya, Jacaratia heterophylla, J. spinosa, Vasconcella cauliflora, V. quercifolia, and V. monoica, all from the family Caricaceae. A fragment of 1,098 nt of the virus isolate, corresponding to the 3' replicase (RdRp) and 5' coat protein (CP) cistron, was cloned and sequenced. Phylogenetic analysis of the RdRp nucleotide sequences indicated some similarity of PLYV with members of the genus Sobemovirus.

Sorologia e sobrevivência do vírus do amarelo letal do mamoeiro

O vírus do amarelo letal do mamoeiro (Papaya lethal yellowing virus, PLYV) é responsável por uma das principais doenças do mamoeiro (Carica papaya) no Nordeste brasileiro. O PLYV pode ser transmitido através do solo, água, instrumento de corte e mãos contaminadas. A presente pesquisa teve como objetivo estudar as características biológicas, sorológicas e físicas de um isolado do vírus e avaliar sua sobrevivência em tecido seco infetado. O PLYV foi detectado por "Enzyme linked immunosorbent assay" (ELISA) indireto e isolado em mudas de mamoeiro, através da inoculação mecânica. A sobrevivência do PLYV em folhas e raízes secas de mamoeiro infetado foi avaliada por sorologia e inoculação em plantas sadias. A presença do vírus foi detectada em folhas e raízes de mamoeiro secas, indicando que o vírus pode permanecer ativo em restos de cultura por até 120 dias. A purificação do PLYV permitiu a obtenção de 309.5 mg de vírus/kg de folha e o anti-soro obtido mostrou-se altamente específico, com títulos de 1:128 em dupla difusão em Agar e 1:1.024.000 em Elisa indireto. Estudos das propriedades físicas do PLYV em mamoeiro revelaram um ponto de inativação térmica (PIT) em torno de 80 ºC, um ponto máximo de diluição (PMD) de ac. 10-6 e uma longevidade in vitro (LIV) acima de 50 dias.

The presence of Papaya ringspot virus and Papaya lethal yellowing virus in commercialized papaya fruits

Considering the high incidence of virus in papaya (Carica papaya) orchards and their typical symptoms in the fruits, a survey was carried out in the Central Market in the State of Ceara (CEASA) for the presence of papaya fruits infected with Papaya ringspot virus (PRSV) and Papaya lethal yellowing virus (PLYV). CEASAs are the most important fruit commercial centers, and a great number of plant products sold in supermarkets are obtained in CEASA. Fruits were randomly collected from CEASA stalls and brought to the Plant Virus Laboratory at the Federal University of Ceara (UFC) to be analyzed for typical virus symptoms and tested by indirect enzyme linked immunosorbent assay (ELISA) against antisera specific to PRSV and PLYV. According to the results obtained by symptom analysis, which were confirmed by serology, among the 8,400 fruits analyzed 5.5% were infected with PRSV and 1.2% with PLYV, showing the need to implement a control program involving eradication of virus sources in the field, represented by infected plants in papaya orchards, abandoned orchards and infected plants identified in backyards.

Transmissão por mãos contaminadas e ausência de transmissão embrionária do vírus do amarelo letal do mamoeiro

O vírus do amarelo letal do mamoeiro ("Papaya lethal yellowing virus", PLYV) é responsável por uma das principais doenças do mamoeiro (Carica papaya) no Nordeste brasileiro. O PLYV pode ser transmitido por solo, água, instrumentos de corte contaminados e por inoculação mecânica em condições experimentais. Na presente pesquisa, avaliou-se a transmissão do PLYV por mãos contaminadas e sua presença em embriões de sementes de frutos infetados. O PLYV foi transmitido por mãos contaminadas, e mãos contaminadas e lavadas em água corrente revelando elevada estabilidade. Experimentos com 1.128 embriões de sementes obtidas de frutos infetados com o vírus revelaram a ausência do PLYV, cuja presença foi constatada em 112 tegumentos das 670 sementes analisadas. O PLYV não foi perpetuado por plantio direto de sementes, envolvendo 310 plântulas e 362 mudas, comprovando não ser o mesmo transmitido por semente de forma embrionária. De outra parte, a presença do PLYV foi comprovada por Elisa indireto em 49 de 456 sementes per si testadas.

Etiologia e estratégias de controle de viroses do mamoeiro no brasil

As viroses constituem o principal grupo de doenças do mamoeiro (Carica papaya), ocasionando grandes perdas na produção, podendo chegar à destruição total das plantações afetadas. Embora mais de dez vírus tenham sido constatados infetando naturalmente o mamoeiro, em todo o mundo, no Brasil, até o presente, foram assinaladas apenas as ocorrências do vírus da mancha anelar do mamoeiro (Papaya ringspot virus, PRSV), do vírus do amarelo letal do mamoeiro (Papaya lethal yellowing virus, PLYV) e do vírus da meleira que se encontra em fase de caracterização. A mancha anelar causada pelo PRSV é, inquestionavelmente, o mais importante problema sanitário do mamoeiro. O controle do PRSV mostra-se imprescindível, apesar de bastante difícil, em razão da sua forma de disseminação rápida e eficiente por diversas espécies de afídeos e ausência de resistência genética em C. papaya. Na tentativa de controlar o PRSV, várias medidas já foram testadas, não existindo, até o momento, nenhuma estratégia eficiente e duradoura para seu controle no Brasil. O desenvolvimento de plantas transgênicas de mamoeiro expressando o gene da capa protéica (cp) do PRSV, imunes ao mesmo, abriu nova possibilidade para solução do problema.

Production of policlonal antisera specific to plant viruses by rabbit oral immunization

Serological techniques are of great importance for plant virus identification and characterization. The major limiting factor for using these techniques for plant virus identification is the requirement of a good virus purified preparation to be used in immunizing animals for antiserum production. In the present study, two New Zealand rabbits were orally immunized with extracts from cowpea (Vigna unguiculata) plants systemically infected with Cowpea severe mosaic virus (CPSMV) and with extracts from papaya (Carica papaya) infected with Papaya lethal yellowing virus (PLYV). The leaf extracts were prepared in saline solution 0.15 M in the rate of 1:1 (w/v) and clarified by a centrifugation of 10,000 g for 10 min. The clarified extracts containing the viruses were orally administered to the New Zealand rabbits in two series of five daily doses of 1.0 ml each. The obtained policlonal antisera were shown to be very specific to their respective viruses in double immunodiffusion and indirect ELISA. These seem to be the first antisera specific for plant virus obtained by rabbit oral immunization. The results open up some possibilities for producing antisera to plant viruses of difficult purification. It is a simple, fast and inexpensive method for production of antisera for plant viruses when compared to the traditional techniques that involve rabbit injections with purified virus preparations.