Sequences of the coat protein gene from brazilian isolates of Papaya ringspot virus

Papaya ringspot virus (PRSV) is the causal agent of the main papaya (Carica papaya) disease in the world. Brazil is currently the world's main papaya grower, responsible for about 40% of the worldwide production. Resistance to PRSV on transgenic plants expressing the PRSV coat protein (cp) gene was shown to be dependent on the sequence homology between the cp transgene expressed in the plant genome and the cp gene from the incoming virus, in an isolate-specific fashion. Therefore, knowledge of the degree of homology among the cp genes from distinct PRSV isolates which are present in a given area is important to guide the development of transgenic papaya for the control of PRSV in that area. The objective of the present study was to assess the degree of homology among the PRSV cp genes of several Brazilian isolates of this virus. Papaya and PRSV are present in many different ecosystems within Brazil. Twelve PRSV isolates, collected in eight different states from four different geographic regions, were used in this study. The sequences of the cp gene from these isolates were compared among themselves and to the gene used to generate transgenic papaya for Brazil. An average degree of homology of 97.3% at the nucleotide sequence was found among the Brazilian isolates. When compared to 27 isolates from outside Brazil in a homology tree, the Brazilian isolates were clustered with Australian, Hawaiian, and Central and North American isolates, with an average degree of homology of 90.7% among them.

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Development of virus resistant transgenic papayas expressing the coat protein gene from a Brazilian isolate of Papaya ringspot virus

Fitopatologia Brasileira ◽

10.1590/s0100-41582005000400004 ◽

2005 ◽

Vol 30 (4) ◽

pp. 357-365 ◽

Cited By ~ 15

Author(s):

Manoel T. Souza Júnior ◽

Osmar Nickel ◽

Dennis Gonsalves

Keyword(s):

Coat Protein ◽

Carica Papaya ◽

Protein Gene ◽

Transformation Efficiency ◽

Ringspot Virus ◽

Zygotic Embryos ◽

Papaya Ringspot Virus ◽

Secondary Somatic Embryo ◽

Embryo Cultures ◽

Transgenic Lines

Translatable and nontranslatable versions of the coat protein (cp) gene of a Papaya ringspot virus (PRSV) isolate collected in the state of Bahia, Brazil, were engineered for expression in Sunrise and Sunset Solo varieties of papaya (Carica papaya). The biolistic system was used to transform secondary somatic embryo cultures derived from immature zygotic embryos. Fifty-four transgenic lines, 26 translatable and 28 nontranslatable gene versions, were regenerated, with a transformation efficiency of 2.7%. Inoculation of cloned R0 plants with PRSV BR, PRSV HA or PRSV TH, Brazilian, Hawaiian and Thai isolates, respectively, revealed lines with mono-, double-, and triple-resistance. After molecular analysis and a preliminary agronomic evaluation, 13 R1 and R2 populations were incorporated into the papaya-breeding program at Embrapa Cassava and Tropical Fruits, in Cruz das Almas, Bahia, Brazil.

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Differential Protection Against Papaya Ringspot Virus Isolates in Coat Protein Gene Transgenic Papaya and Classically Cross-Protected Papaya

Phytopathology ◽

10.1094/phyto-84-1359 ◽

1994 ◽

Vol 84 (11) ◽

pp. 1359 ◽

Cited By ~ 97

Author(s):

P. F. Tennant

Keyword(s):

Coat Protein ◽

Coat Protein Gene ◽

Protein Gene ◽

Ringspot Virus ◽

Differential Protection ◽

Papaya Ringspot Virus ◽

Transgenic Papaya ◽

Virus Isolates

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Comparative reactions of recombinant papaya ringspot viruses with chimeric coat protein (CP) genes and wild-type viruses on CP-transgenic papaya

Journal of General Virology ◽

10.1099/0022-1317-82-11-2827 ◽

2001 ◽

Vol 82 (11) ◽

pp. 2827-2836 ◽

Cited By ~ 32

Author(s):

Chu-Hui Chiang ◽

Ju-Jung Wang ◽

Fuh-Jyh Jan ◽

Shyi-Dong Yeh ◽

Dennis Gonsalves

Keyword(s):

Coat Protein ◽

Sequence Similarity ◽

Transcriptional Gene Silencing ◽

Papaya Ringspot Virus ◽

Wild Type ◽

Coding Region ◽

Transgenic Papaya ◽

Cp Gene ◽

Post Transcriptional Gene Silencing

Transgenic papaya cultivars SunUp and Rainbow express the coat protein (CP) gene of the mild mutant of papaya ringspot virus (PRSV) HA. Both cultivars are resistant to PRSV HA and other Hawaii isolates through homology-dependent resistance via post-transcriptional gene silencing. However, Rainbow, which is hemizygous for the CP gene, is susceptible to PRSV isolates from outside Hawaii, while the CP-homozygous SunUp is resistant to most isolates but susceptible to the YK isolate from Taiwan. To investigate the role of CP sequence similarity in overcoming the resistance of Rainbow, PRSV HA recombinants with various CP segments of the YK isolate were constructed and evaluated on Rainbow, SunUp and non-transgenic papaya. Non-transgenic papaya were severely infected by all recombinants, but Rainbow plants developed a variety of symptoms. On Rainbow, a recombinant with the entire CP gene of YK caused severe symptoms, while recombinants with only partial YK CP sequences produced a range of milder symptoms. Interestingly, a recombinant with a YK segment from the 5′ region of the CP gene caused very mild, transient symptoms, whereas recombinants with YK segments from the middle and 3′ parts of the CP gene caused prominent and lasting symptoms. SunUp was resistant to all but two recombinants, which contained the entire CP gene or the central and 3′-end regions of the CP gene and the 3′ non-coding region of YK, and the resulting symptoms were mild. It is concluded that the position of the heterologous sequences in the recombinants influences their pathogenicity on Rainbow.

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Engineered Resistance Against Papaya ringspot virus in Venezuelan Transgenic Papayas

Plant Disease ◽

10.1094/pdis.2004.88.5.516 ◽

2004 ◽

Vol 88 (5) ◽

pp. 516-522 ◽

Cited By ~ 28

Author(s):

Gustavo Fermin ◽

Valentina Inglessis ◽

Cesar Garboza ◽

Sairo Rangel ◽

Manuel Dagert ◽

...

Keyword(s):

Amino Acid ◽

Agrobacterium Tumefaciens ◽

Coat Protein ◽

Amino Acid Sequence ◽

Sequence Similarity ◽

Ringspot Virus ◽

Amino Acid Sequence Similarity ◽

Papaya Ringspot Virus ◽

Local Varieties ◽

Cp Gene

Local varieties of papaya grown in the Andean foothills of Mérida, Venezuela, were transformed independently with the coat protein (CP) gene from two different geographical Papaya ringspot virus (PRSV) isolates, designated VE and LA, via Agrobacterium tumefaciens. The CP genes of both PRSV isolates show 92 and 96% nucleotide and amino acid sequence similarity, respectively. Four PRSV-resistant R0 plants were intercrossed or selfed, and the progenies were tested for resistance against the homologous isolates VE and LA, and the heterologous isolates HA (Hawaii) and TH (Thailand) in greenhouse conditions. Resistance was affected by sequence similarity between the transgenes and the challenge viruses: resistance values were higher for plants challenged with the homologous isolates (92 to 100% similarity) than with the Hawaiian (94% similarity) and, lastly, Thailand isolates (88 to 89% similarity). Our results show that PRSV CP gene effectively protects local varieties of papaya against homologous and heterologous isolates of PRSV.

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Virus Resistant Papaya Plants Derived from Tissues Bombarded with the Coat Protein Gene of Papaya Ringspot Virus

Nature Biotechnology ◽

10.1038/nbt1192-1466 ◽

1992 ◽

Vol 10 (11) ◽

pp. 1466-1472 ◽

Cited By ~ 145

Author(s):

Maureen M. M. Fitch ◽

Richard M. Manshardt ◽

Dennis Gonsalves ◽

Jerry L. Slightom ◽

John C. Sanford

Keyword(s):

Coat Protein ◽

Coat Protein Gene ◽

Protein Gene ◽

Ringspot Virus ◽

Papaya Ringspot Virus

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Stability of PRV Resistance in Transgenic Papaya

HortScience ◽

10.21273/hortsci.30.4.904a ◽

1995 ◽

Vol 30 (4) ◽

pp. 904A-904

Author(s):

R. Manshardt ◽

S. Lius ◽

D. Gonsalves ◽

M. Fitch ◽

J. Slightom ◽

...

Keyword(s):

Coat Protein ◽

Coat Protein Gene ◽

Transgene Expression ◽

Protein Gene ◽

Papaya Ringspot Virus ◽

Mendelian Segregation ◽

Transgenic Papaya ◽

Aphid Inoculation ◽

High Level ◽

Level Resistance

Transgenic papaya lines carrying the coat protein gene (CP) of papaya ringspot virus (PRV) strain HA 5-1 display PRV reactions ranging from complete susceptibility (39-3 & 39-4), to slight delay in onset of symptoms (39-1) and attenuation of symptoms (60-3), to high-level resistance (55-1, 63-1). Normal Mendelian segregation of transgene expression was lost in R1 of 39-3 and 39-4, and inbred R1 60-3 gave an aberrant 1:1 ratio. R0 55-1 plants were resistant in the field (Hawaii) for 2 years following manual and/or aphid inoculation, and the high-level resistance remained stable in the R1 after repeated manual inoculations in the greenhouse and graft inoculation for up to 1 year (Cornell). However, inoculation with PRV HA-Oahu strain produced symptoms in some plants at Cornell (9% after 6 weeks) and in Hawaii (50% after 1 year). Two 55-1 and one 60-3 plant subsequently underwent remission of symptoms and became ELISA-negative (Hawaii). Transmission of PRV isolates from symptomatic 55-1 plants to other CP+ 55-1 bioassay plants was unsuccessful.

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RELATIONSHIP BETWEEN PAPAYA RINGSPOT VIRUS COAT PROTEIN TRANSGENE EXPRESSION LEVELS AND AGE-DEPENDENT RESISTANCE IN TRANSGENIC PAPAYA

Acta Horticulturae ◽

10.17660/actahortic.2002.575.5 ◽

2002 ◽

pp. 75-83 ◽

Cited By ~ 1

Author(s):

D.A. Gaskill ◽

K.Y. Pitz ◽

S.A. Ferreira ◽

D. Gonsalves

Keyword(s):

Coat Protein ◽

Transgene Expression ◽

Ringspot Virus ◽

Papaya Ringspot Virus ◽

Transgenic Papaya ◽

Expression Levels ◽

Virus Coat Protein ◽

Age Dependent ◽

Virus Coat

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Genetic variability of Papaya ringspot virus isolates in Norte de Santander - Colombia

Agronomía Colombiana ◽

10.15446/agron.colomb.v33n2.50095 ◽

2015 ◽

Vol 33 (2) ◽

pp. 184-193 ◽

Cited By ~ 3

Author(s):

Giovanni Chaves-Bedoya ◽

Luz Yineth Ortiz-Rojas

Keyword(s):

Coat Protein ◽

Genetic Variability ◽

Carica Papaya ◽

Local Level ◽

Bioinformatic Analysis ◽

Ringspot Virus ◽

Limiting Factor ◽

Papaya Ringspot Virus ◽

Phylogenetic Groups ◽

Valle Del Cauca

The Papaya ringspot virus (PRSV), a member of the potyvirus that is transmitted by aphids within the Potyviridae family, is the main limiting factor for papaya (Carica papaya L.) and Cucurbits worldwide and causes losses of up to 100%. In this study, we conducted research on the genetic diversity of PRSV isolates collected from two locations in the department of Norte de Santander, Colombia. The analysis was performed by comparing the nucleotide sequences of the region that encode the coat protein (CP) of nine PRSV isolates from the Villa del Rosario location and 12 isolates from the Campo Hermoso location. The analysis included three sequences of the CP of PRSV isolates reported in the Colombian departments of Arauca and Valle del Cauca. The bioinformatic analysis suggested that the PRSV isolates from the locations in Norte de Santander were different from each other, grouping into different phylogenetic groups. Anexistence of recombination events in the coat protein was observed. This is the first study of PRSV genetic variability that has been conducted at the local level in Colombia.

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