Home > XIX International Symposium on Virus and Virus-like Diseases of Temperate Fruit Crops – Fruit Tree Diseases > EXPRESSION OF SINGLE-CHAIN VARIABLE FRAGMENTS AGAINST STRUCTURAL AND NON-STRUCTURAL PROTEINS OF PLUM POX VIRUS IN TRANSGENIC NICOTIANA BENTHAMIANA PLANTS TO INTERFERE WITH VIRAL INFECTION

Articles

EXPRESSION OF SINGLE-CHAIN VARIABLE FRAGMENTS AGAINST STRUCTURAL AND NON-STRUCTURAL PROTEINS OF PLUM POX VIRUS IN TRANSGENIC NICOTIANA BENTHAMIANA PLANTS TO INTERFERE WITH VIRAL INFECTION

Article number

657_54

Pages

341 – 347

Language

English

Abstract

Single chain variable fragments (scFv) specific to the coat protein (CP) (scFv4C) and the RNA replicase NIb (scFv2A) from Plum pox virus (PPV) were engineered.
Diverse expression versions addressed to different cell compartments were produced by addition of several peptides.
Two cytosolic versions of scFv4C with and without KDEL tetrapeptide (scFv4CKDEL) were generated, as well as, three scFv2A versions: one cytosolic (scFv2A), one nuclear (NLS-scFv2A), and a cytosolic facing endoplasmic reticulum membrane targeted version (6K2-scFv2A). Transgenic Nicotiana benthamiana plants expressing these different scFv versions were obtained and some lines from each construct were selected for viral challenge.
Challenge experiments were performed by mechanical inoculation with a PPV-D/NAT isolate at moderate inoculum doses (< 0.36 μg/ml viral concentration). The percentage of infected plants was significantly lower in some transgenic lines compared to control plants.
Nuclear version of scFv2A and cytosolic version with KDEL of scFv4C showed the highest reduction in viral infection.

Publication

XIX International Symposium on Virus and Virus-like Diseases of Temperate Fruit Crops – Fruit Tree Diseases

Authors

M. Gil, O. Esteban, M.C. Martínez, M.T. Gorris, L. Peña, M. Cambra, J.A. García

Keywords

Single-chain antibody, Plum pox virus, viral resistance, NIb replicase, coat protein, transgenic plants

Full text

https://doi.org/10.17660/ActaHortic.2004.657.54

Online Articles (94)

2. PRODUCTION AND USE OF POLYCLONAL ANTIBODIES TO THE COAT PROTEIN OF APPLE STEM GROOVING VIRUS EXPRESSED IN ESCHERICHIA COLI

O. Nickel | T.V. Fajardo | A.P. Trivilin | M.L.N.P. Targon | M.A. Machado

3. STUDIES ON THE INFLUENCE OF APPLE STEM GROOVING VIRUS ON TREE GROWTH OF VARIOUS APPLE CULTIVARS IN THE NURSERY

A. Maxim | L. Zagrai | I. Zagrai | M. Isac

4. APPLE STEM PITTING VIRUS DETECTION FROM DORMANT POME FRUITS BY RT-PCR

S. Paunović | D. Jevremović

5. APPLE VIRAL DISEASES DIAGNOSED IN THE GERMPLASM COLLECTION AT I.C.D.P. MARACINENI-ROMANIA

S. Popescu | G. Constantin | CR. Mazilu

6. A PRELIMINARY ACCOUNT OF THE PRESENCE OF POME FRUIT VIRUSES IN ALBANIA

7. PRODUCTION OF MONOCLONAL ANTIBODIES FOR THE SEROLOGICAL IDENTIFICATION AND RELIABLE DETECTION OF APPLE STEM PITTING AND PEAR YELLOW VEIN VIRUSES IN APPLE AND PEAR

P. Gugerli | M.E. Ramel

8. COMPARATIVE RESULTS BETWEEN DIFFERENT DETECTION METHODS OF VIRUS AND PHYTOPLASMAS FOR A PEAR AND APPLE CERTIFICATION PROGRAM

9. IDENTIFICATION OF A NEW TRICHOVIRUS IN SOME FRUIT TREES INFECTED BY APPLE CHLOROTIC LEAF SPOT VIRUS (ACLSV)

10. MOLECULAR CHARACTERIZATION OF A NEW FOVEAVIRUS IN PRUNUS ACCESSIONS OF ASIAN ORIGIN

A. Marais | L. Svanella-Dumas | X. Foissac | T. Candresse

11. DETECTION OF PLUM BARK NECROSIS STEM PITTING-ASSOCIATED VIRUS (PBNSPAV) FROM DIFFERENT STONE FRUIT SPECIES AND OPTIMISATION OF DIAGNOSTIC TOOLS

T. Amenduni | C. Hobeika | A. Minafra | V. Savino

12. NUCLEOTIDE SEQUENCE ANALYSIS AND DETECTION OF CHERRY RASP LEAF VIRUS

D. James

13. BIOLOGICAL PROPERTIES AND PARTIAL MOLECULAR CHARACTERIZATION OF AN APRICOT STRAIN OF CGRMV

14. VERTICAL TRANSMISSION OF PRUNUS NECROTIC RINGSPOT VIRUS BY GAMETES IN APRICOT

K. Amari | M.A. Sánchez-Pina | V. Pallás

15. THE OCCURRENCE AND MOLECULAR CHARACTERISATION OF PNRSV ISOLATES IN TURKEY

C. Ulubas | F. Ertunc

16. APPLE CHLOROTIC LEAF SPOT VIRUS INFECTION INDUCES PLUM POX VIRUS-LIKE SYMPTOMS ON PLUM IN NEW ZEALAND

17. DETECTION OF SOME VIRUSES OF STONE FRUITS IN MOTHER PLANT BLOCKS IN EASTERN MEDITERRANEAN REGION OF TURKEY

G. Sertkaya | K. Caglayan | C. Ulubas

18. DETECTION AND QUANTIFICATION OF PLUM POX POTYVIRUS IN APHID VECTORS BY REAL-TIME FLUORESCENT REVERSE TRANSCRIPTION-PCR

W.L. Schneider | D.J. Sherman | A.L. Stone | V.D. Damsteegt | R.D. Frederick

19. REAL-TIME RT-PCR OF PPV WITH R.A.P.I.D. A FIELD-HARDENED PCR UNIT FOR IN-FIELD DETECTION

V.A. Mavrodieva | L. Levy

20. REAL-TIME RT-PCR FOR QUANTITATIVE DETECTION OF PLUM POX VIRUS

A. Olmos | E. Bertolini | M. Gil | M. Cambra

21. DIRECT NESTED RT-PCR FOR THE DETECTION OF PLUM POX VIRUS

C. Varveri

22. FIRST DETECTION AND MOLECULAR CHARACTERISATION OF PPV-M STRAINS IN PLUM ORCHARDS IN SOUTH-WESTERN GERMANY

W. Jarausch | A. Baβler | N. Molla | G. Krczal

23. DETECTION AND CHARACTERIZATION OF PLUM POX VIRUS ISOLATES IN LOMBARDIA AND VENETO REGIONS (ITALY)

24. STRATEGY TO IDENTIFY AND TO CHARACTERIZE NEW ISOLATES OF PLUM POX VIRUS

M. Ravelonandro | P. Briard | M. Monsion

25. PRELIMINARY MOLECULAR CHARACTERIZATION OF PLUM POX POTYVIRUS ISOLATE W3174: EVIDENCE OF A NEW STRAIN

D. James | A. Varga

26. CHARACTERIZATION OF PLUM POX POTYVIRUS (PPV) BY DAS-ELISA AND RT-PCR/RFLP ANALYSIS IN TURKEY

K. Caglayan | G. Sertkaya | C. Ulubas | M. Kölber | L. Krizbai

27. PLUM POX VIRUS IN TUNISIA: SOURCES AND PRELIMINARY CHARACTERISTICS

M. Boulila | P. Briard | M. Ravelonandro

28. EPIDEMIOLOGY OF PLUM POX VIRUS IN JAPANESE PLUMS IN SPAIN

29. POTENTIAL PRUNUS HOST RANGE OF PPV-PENN ISOLATES BY APHID TRANSMISSION

V.D. Damsteegt | A.L. Stone | W.L. Schneider | D.G. Luster | F.E. Gildow

30. TRANSMISSION OF THREE NORTH AMERICAN ISOLATES OF PLUM POX VIRUS: IDENTIFICATION OF APHID VECTORS AND SPECIES-SPECIFIC TRANSMISSION FROM INFECTED STONE FRUITS

31. SPATIALLY ARRANGED DIVERSITY OF PLUM POX VIRUS (PPV) ISOLATES IN A CLUSTER OF ORCHARDS IN SOUTHERN FRANCE

V. Marie-Jeanne | G. Thébaud | J. Peyre | G. Labonne

32. APHID TRANSMISSION OF NATURAL RECOMBINANT PLUM POX VIRUS ISOLATES TO DIFFERENT PRUNUS SPP.-A CONTRIBUTION FOR UNDERSTANDING THE EPIDEMIOLOGY OF AN ATYPICAL PPV

M. Glasa | M. Boeglin | G. Labonne

33. RISK ASSESSMENT OF CONTAMINATION OF CHERRY TREES BY PLUM POX VIRUS IN FRANCE

M. Boeglin | J.B. Quiot | G. Labonne

34. SOUR CHERRY ISOLATE OF PLUM POX VIRUS (PPV-SOC): MONITORING OF VIRAL DISTRIBUTION IN INFECTED PRUNUS ROOTSTOCKS

M. Bodin | D. Verger | E. Costes | F. Dosba | M. Glasa

35. DETECTION AND ERADICATION OF PLUM POX VIRUS MARCUS TYPE IN ARAGÓN (SPAIN)

36. THE OCCURRENCE OF PPV IN CHERRY TREES IN THE CZECH REPUBLIC

37. CURRENT STATUS OF SHARKA DISEASE IN SLOVENIA

M. Virček | I. Mavrič

38. DETECTION OF PLUM POX POTYVIRUS IN WEEDS

M. Virček | I. Mavrič | M. Urbančič-Zemljič | V. kerlavaj

39. EVALUATION OF THREE ORNAMENTAL PRUNUS AS RESERVOIRS OF PPV

G. Labonne | M. Boeglin | B. Monsion

40. VARIABILITY IN SUSCEPTIBILITY TO PLUM POX VIRUS IN NATURAL WOODY HOSTS, MYROBALAN AND BLACKTHORN

J. Polák

41. RELATIONSHIP BETWEEN PLUM POX VIRUS AND PLUM BARK SPLIT VIRUS (ACLSV) IN PLUM

N. Minoiu | D. Vladianu | I. Oprean | M. Ravelonandro | P. Stegerean

42. IDENTIFICATION OF PLUM POX VIRUS AND SOME ILAR VIRUSES OF STONE FRUITS IN KAHRAMANMARAS DISTRICT OF TURKEY

E. Tolay Arikan | K. Caglayan | M.H. Gazel

43. RESISTANCE TO PLUM POX VIRUS: A MOLECULAR APPROACH

44. MOLECULAR GENETIC MAPPING OF THE PLUM POX VIRUS RESISTANCE GENES IN APRICOT

45. UTILIZATION OF ALMOND AS SOURCE OF PLUM POX VIRUS RESISTANCE IN PEACH BREEDING

P. Martínez-Gómez | M. Rubio | F. Dicenta | T.M. Gradziel

46. LOCALIZATION OF COAT PROTEIN AND NUCLEIC ACID OF PLUM POX VIRUS IN PRUNUS PETIOLE AND STEM TISSUES

47. IN SITU LOCALISATION OF PLUM POX PARTICLES IN SUSCEPTIBLE AND RESISTANT APRICOT CULTIVARS

48. EVALUATION OF THE SUSCEPTIBILITY OF SEEDLINGS FROM APRICOT CROSSES TO THE MARCUS STRAIN OF PLUM POX VIRUS

T. Amenduni | A. Bazzoni | A. Minafra | V. Savino

49. EVALUATION OF CHERRY CULTIVARS FOR THEIR RESPONSE TO INFECTION BY PLUM POX VIRUS SWEET CHERRY STRAIN

A. Fanigliulo | S. Comes | A. Crescenzi

50. ANALYSIS OF METHYLATION AND POST-TRANSCRIPTIONAL GENE SILENCING IN EMBRYOS AND SEEDLINGS OF C5, A TRANSGENIC PLUM POX VIRUS RESISTANT PLUM PRUNUS DOMESTICA L.

R. Scorza | K. Webb | M. Ravelonandro

51. POTENTIAL USE OF TRANSGENIC PLUMS RESISTANT TO PLUM POX VIRUS FIELD INFECTION

R. Scorza | T. Malinowski | N. Minoiu | M. Ravelonandro | M. Cambra

52. INVESTIGATIONS OF POTENTIAL ENVIRONMENTAL IMPACTS IN THE RELEASE OF TRANSGENIC PLUMS

M. Ravelonandro | N. Minoiu | R. Scorza

53. PRUNUS/PPV INTERACTIONS: A MOLECULAR APPROACH

54. THE USE OF GREEN FLUORESCENT PROTEIN-TAGGED RECOMBINANT PLUM POX VIRUS (PPV) TO STUDY VIRAL INVASION IN PPV/PRUNUS INTERACTION

55. EXPRESSION OF SINGLE-CHAIN VARIABLE FRAGMENTS AGAINST STRUCTURAL AND NON-STRUCTURAL PROTEINS OF PLUM POX VIRUS IN TRANSGENIC NICOTIANA BENTHAMIANA PLANTS TO INTERFERE WITH VIRAL INFECTION

56. EFFECT OF PLUM POX VIRUS INFECTION ON PHOTOSYNTHESIS AND ANTIOXIDANT ENZYME ACTIVITY IN PEACH ‘GF305’

57. EXPERIMENTAL EVIDENCE THAT APPLE DIMPLE FRUIT VIROID DOES NOT SPREAD NATURALLY

M. Malfitano | D. Alioto | A. Ragozzino | F. Di Serio | R. Flores

58. DETECTION OF APPLE SCAR SKIN VIROID IN APPLE TREES BY THE ISOTHERMAL NUCLEIC ACID AMPLIFICATION AND ELECTROCHEMILUMINESCENCE

59. IDENTIFICATION AND MOLECULAR CHARACTERIZATION OF PEAR BLISTER CANKER VIROID ISOLATES IN CAMPANIA (SOUTHERN ITALY)

M. Malfitano | M. Barone | E. Ragozzino | D. Alioto | R. Flores

60. DETECTION OF PEAR BLISTER CANKER VIROID IN POME FRUIT ORCHARDS ACROSS AUSTRALIA

P. Joyce | B. Rodoni | J.M. Crosslin | W.E. Howell | K.C. Eastwell

61. DETECTION BY TISSUE PRINTING OF STONE FRUIT VIROIDS, FROM EUROPE, THE MEDITERRANEAN AND NORTH AND SOUTH AMERICA

62. MOLECULAR CHARACTERIZATION OF HOP STUNT VIROID (HSVD) SEQUENCE VARIANTS FROM PRUNUS SPECIES

E. Ragozzino | F. Faggioli | M. Barba | M. Malfitano

63. DEVELOPMENT OF A PCR METHOD OF PEACH LATENT MOSAIC VIROID AND HOP STUNT VIROID DETECTION FOR CERTIFICATION OF PLANTING MATERIAL

M. Hassan | M. Zouhar | P. Rysanek

64. MIXED INFECTION OF PEACH TREES WITH PEACH LATENT MOSAIC VIROID AND HOP STUNT VIROID IN THE CZECH REPUBLIC

M. Hassan | P. Rysanek | F. Di Serio

65. INTERACTIONS BETWEEN CITRUS CACHEXIA VIROID AND CLOSELY RELATED SEQUENCE VARIANTS MAY IMPAIR EXPRESSION OF CACHEXIA SYMPTOMS

66. OVERWINTERING ADULTS AND SPRINGTIME GENERATION OF CACOPSYLLA PICTA (SYNONYM C. COSTALIS) CAN TRANSMIT APPLE PROLIFERATION PHYTOPLASMAS

B. Jarausch | N. Schwind | W. Jarausch | G. Krczal

67. ESTABLISHMENT OF A QUANTITATIVE REAL-TIME PCR ASSAY FOR THE QUANTIFICATION OF APPLE PROLIFERATION PHYTOPLASMAS IN PLANTS AND INSECTS

W. Jarausch | T. Peccerella | N. Schwind | B. Jarausch | G. Krczal

68. ANALYSIS OF THE DISTRIBUTION OF APPLE PROLIFERATION PHYTOPLASMA SUBTYPES IN A LOCAL FRUIT GROWING REGION IN SOUTHWEST GERMANY

W. Jarausch | N. Schwind | B. Jarausch | G. Krczal

69. GENETIC VARIABILITY OF AP PHYTOPLASMAS DETECTED IN THE APPLE GROWING AREA OF TRENTINO (NORTH ITALY)

C. Cainelli | C. Bisognin | M.E. Vindimian | M.S. Grando

70. PEAR DECLINE PHYTOPLASMA DETECTION IN PEAR TREES AND INSECT VECTORS BY DOT-BLOT HYBRIDIZATION AND NESTED-PCR

M. García-Chapa | A. Batlle | A. Laviña | L. Galipienso | G. Firrao

71. TOLERANCE INCREASE TO PEAR DECLINE PHYTOPLASMA IN MYCORRHIZAL OHF-333 PEAR ROOTSTOCK

72. IDENTIFICATION OF PHYTOPLASMAS ON POMACEOUS FRUIT TREE SPECIES IN HUNGARY

73. TRANSMISSION OF EUROPEAN STONE FRUIT YELLOWS PHYTOPLASMA TO PRUNUS SPECIES BY USING VECTOR AND GRAFT TRANSMISSION

L. Carraro | F. Ferrini | P. Ermacora | N. Loi

74. DISTRIBUTION OF EUROPEAN STONE FRUIT YELLOWS PHYTOPLASMA IN APRICOT TREES IN WESTERN SWITZERLAND

M. Genini | M.E. Ramel

75. EPIDEMIOLOGICAL SURVEY OF EUROPEAN STONE FRUIT YELLOWS PHYTOPLASMA IN TWO ORCHARDS IN WESTERN SWITZERLAND

M.E. Ramel | P. Gugerli

76. DATA ON THE LIFE CYCLE OF CACOPSYLLA PRUNI, PSYLLIDAE VECTOR OF EUROPEAN STONE FRUIT YELLOWS (ESFY) PHYTOPLASMA, IN FRANCE

G. Labonne | J. Lichou

77. SPATIO-TEMPORAL ANALYSIS OF DISEASE SPREAD PROVIDES INSIGHTS INTO THE EPIDEMIOLOGY OF EUROPEAN STONE FRUIT YELLOWS

G. Thébaud | C. Castelain | G. Labonne | J. Chadoeuf

78. SURVIVAL OF EUROPEAN STONE FRUIT YELLOWS PHYTOPLASMA OUTSIDE FRUIT CROP PRODUCTION AREAS: A CASE STUDY IN SOUTHEASTERN FRANCE

M. Yvon | G. Labonne | G. Thébaud

79. EPIDEMIOLOGY OF EUROPEAN STONE FRUIT YELLOWS PHYTOPLASMA IN THE CZECH REPUBLIC

80. OCCURRENCE AND EPIDEMIOLOGY OF EUROPEAN STONE FRUIT YELLOWS PHYTOPLASMA IN SPAIN

A. Laviña | J. Sabaté | M. García-Chapa | A. Batlle | E. Torres

81. IMPROVED MOLECULAR METHODS FOR DETECTION OF EUROPEAN STONE FRUIT YELLOWS (ESFY) PHYTOPLASMAS FROM IN VITRO SHOOTS OF FRUIT TREES

82. THREE YEARS OF MOLECULAR MONITORING OF PHYTOPLASMA SPREADING IN A PLUM GROWING AREA IN ITALY

S. Paltrinieri | A. Bertaccini | A. Lugli | W. Monari

83. PHYTOPLASMA DETECTION IN EMPOASCA DECEDENS AND EMPOASCA SPP. AND THEIR POSSIBLE ROLE AS VECTORS OF EUROPEAN STONE FRUIT YELLOWS (16SRX-B) PHYTOPLASMA

84. DETECTION OF EUROPEAN STONE FRUIT YELLOWS PHYTOPLASMA (ESFYP) IN HOMOPTERA INSECTS AND IN WILD STONE FRUIT TREES COLLECTED IN PEACH ORCHARDS IN NORTHERN ITALY

85. FIRST DETECTION OF EUROPEAN STONE FRUIT YELLOWS PHYTOPLASMA (ESFY) IN BELGIUM

T. Olivier | J. Kummert | S. Steyer

86. PHYTOPLASMA DISEASES OF STONE FRUIT TREES IN POLAND

M. Cieślińska | H. Morgaś | T. Jakubowski

87. PHYLOGENETICAL CHARACTERIZATION AND PCR DETECTION OF A NEW PHYTOPLASMA IN ALMOND (PRUNUS AMYGDALUS) AND PEACH (PRUNUS PERSICAE) IN THE MEDITERRANEAN AREA

88. OCCURRENCE OF PSYLLIDS IN APPLE ORCHARDS AND TRANSMISSION OF APPLE PROLIFERATION BY CACOPSYLLA PICTA (SYN. C. COSTALIS) IN GERMANY

E. Seemüller | E. Dickler | C. Berwarth | W. Jelkmann

89. SAMPLING PROTOCOLS AND RISK OF ERROR SIGNIFICANCE IN MOLECULAR DETECTION TESTS FOR FRUIT TREES CERTIFICATION

90. DEVELOPMENT OF RT-PCR ASSAYS USING FLUOROGENIC 3′ MINOR GROOVE BINDER DNA PROBES FOR DETECTION OF FRUIT TREE VIRUSES

91. MULTIPLEX DETECTION OF PLANT (QUARANTINE) PATHOGENS BY MICRO-ARRAYS: AN INNOVATIVE TOOL FOR PLANT HEALTH MANAGEMENT

92. NEW APPROACH IN THE IDENTIFICATION OF THE CAUSAL AGENT OF FIG MOSAIC DISEASE

93. IDENTIFICATION OF CITRUS TRISTEZA VIRUS STRAINS IN LOUISIANA AND ALABAMA

94. INTERNATIONAL WORKING GROUP ON FRUIT TREE VIRUSES

W. Jelkmann