A cell wall-localized NLR confers resistance to Soybean mosaic virus by recognizing viral-encoded cylindrical inclusion protein

Potato virus A (PVA) particles were purified by centrifugation through a 30 % sucrose cushion and the pellet (P1) was resuspended and sedimented through a 5–40 % sucrose gradient. The gradient separation resulted in two different virus particle populations: a virus fraction (F) that formed a band in the gradient and one that formed a pellet (P2) at the bottom of the gradient. All three preparations contained infectious particles that retained their integrity when visualized by electron microscopy (EM). Western blotting of the P1 particles revealed that the viral RNA helicase, cylindrical inclusion protein (CI), co-purified with virus particles. This result was confirmed with co-immunoprecipitation experiments. CI was detected in P2 particle preparations, whereas F particles were devoid of detectable amounts of CI. ATPase activity was detected in all three preparations with the greatest amount in P2. Results from immunogold-labelling EM experiments suggested that a fraction of the CI present in the preparations was localized to one end of the virion. Atomic force microscopy (AFM) studies showed that P1 and P2 contained intact particles, some of which had a protruding tip structure at one end, whilst F virions were less stable and mostly appeared as beaded structures under the conditions of AFM. The RNA of the particles in F was translated five to ten times more efficiently than RNA from P2 particles when these preparations were subjected to translation in wheat-germ extracts. The results are discussed in the context of a model for CI-mediated functions.

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Strain-Specific Cylindrical Inclusion Protein of Soybean mosaic virus Elicits Extreme Resistance and a Lethal Systemic Hypersensitive Response in Two Resistant Soybean Cultivars

Molecular Plant-Microbe Interactions ◽

10.1094/mpmi-22-9-1151 ◽

2009 ◽

Vol 22 (9) ◽

pp. 1151-1159 ◽

Cited By ~ 42

Author(s):

Jang-Kyun Seo ◽

Suk-Ha Lee ◽

Kook-Hyung Kim

Keyword(s):

Amino Acid ◽

Mosaic Virus ◽

Amino Acid Substitution ◽

Hypersensitive Response ◽

Soybean Mosaic Virus ◽

Cylindrical Inclusion ◽

Single Amino Acid ◽

Extreme Resistance ◽

Soybean Genotypes ◽

Pathogenic Determinant

In the Soybean mosaic virus (SMV)–soybean pathosystem, three independent genes (Rsv1, Rsv3, and Rsv4) conferring resistance to SMV have been identified. Recently, we constructed infectious cDNA clones of SMV G7H and G5H strains and found that these two strains differ in their ability to infect soybean genotypes possessing different SMV resistance genes despite a difference of only 33 amino acids. In particular, pSMV-G7H induced mosaic symptoms systemically in L29 (Rsv3) and provoked a lethal systemic hypersensitive response (LSHR) in Jinpumkong-2, whereas pSMV-G5H could not infect these soybean genotypes. To identify the responsible pathogenic determinants of SMV, we exploited the differential responses of pSMV-G7H- and pSMV-G5H-derived chimeric viruses and amino acid substitution mutant viruses in several soybean genotypes and demonstrated that cylindrical inclusion (CI) protein is the elicitor of Rsv3-mediated extreme resistance and a pathogenic determinant provoking LSHR in Jinpumkong-2. A single amino acid substitution in CI was found to be responsible for gain or loss of elicitor function of CI. Our finding provides a role for CI as a pathogenic determinant in the SMV–soybean pathosystem, and increases the understanding of the basis of the different disease responses of SMV strains.

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Detection of the movement protein of red clover necrotic mosaic virus in a cell wall fraction from infected Nicotiana clevelandii plants

Journal of General Virology ◽

10.1099/0022-1317-72-11-2853 ◽

1991 ◽

Vol 72 (11) ◽

pp. 2853-2856 ◽

Cited By ~ 17

Author(s):

T. A. M. Osman ◽

K. W. Buck

Keyword(s):

Cell Wall ◽

Mosaic Virus ◽

Movement Protein ◽

Red Clover ◽

Cell Wall Fraction ◽

A Cell ◽

Nicotiana Clevelandii

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The roles of the cylindrical inclusion protein of a potyvirus in the induction of vesicles and in cell-to-cell spread

Journal of Structural Biology ◽

10.1016/1047-8477(90)90099-x ◽

1990 ◽

Vol 105 (1-3) ◽

pp. 62-66 ◽

Cited By ~ 8

Author(s):

Victoria L. Calder ◽

Manfred Ingerfeld

Keyword(s):

Cylindrical Inclusion ◽

Inclusion Protein ◽

Cylindrical Inclusion Protein ◽

Cell Spread

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Sequential appearance of capsid protein and cylindrical inclusion protein in root-tip cells following infection with passion fruit woodiness virus

Journal of Ultrastructure and Molecular Structure Research ◽

10.1016/0889-1605(88)90037-7 ◽

1988 ◽

Vol 100 (3) ◽

pp. 201-211 ◽

Cited By ~ 2

Author(s):

Na-Sheng Lin ◽

Nancy Wang ◽

Yau-Heiu Hsu

Keyword(s):

Capsid Protein ◽

Passion Fruit ◽

Cylindrical Inclusion ◽

Root Tip ◽

Inclusion Protein ◽

Cylindrical Inclusion Protein ◽

Tip Cells ◽

Root Tip Cells ◽

Passion Fruit Woodiness Virus ◽

Sequential Appearance

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Identification of Soybean mosaic virus Strains by RT-PCR/RFLP Analysis of Cylindrical Inclusion Coding Region

Plant Disease ◽

10.1094/pdis.2004.88.6.641 ◽

2004 ◽

Vol 88 (6) ◽

pp. 641-644 ◽

Cited By ~ 9

Author(s):

Yul-Ho Kim ◽

Ok-Sun Kim ◽

Jae-Hwan Roh ◽

Jung-Kyung Moon ◽

Soo-In Sohn ◽

...

Keyword(s):

Mosaic Virus ◽

Soybean Mosaic Virus ◽

Rflp Analysis ◽

Cylindrical Inclusion ◽

Enzyme Linked Immunosorbent Assay ◽

Strain Identification ◽

Coding Region ◽

Rt Pcr ◽

Detection And Identification ◽

Pcr Rflp

A reverse-transcriptase polymerase chain reaction/restriction fragment length polymorphism (RT-PCR/RFLP) was employed successfully for detection and identification of Soybean mosaic virus (SMV) strains. A primer pair amplifying a 1,385-bp fragment of the cylindrical inclusion (CI) coding region was used for RT-PCR and the RFLP profiles of the RT-PCR products were compared after restriction digestion with RsaI, EcoRI, or AccI restriction endonucleases. These enzymes were chosen based on the nucleotide sequences of SMV strains G2, G5, G5H, G7, and G7H in the CI coding region. These five strains, as well as seedborne SMV isolates from local soybean cultivars, could be differentiated by RT-PCR/RFLP analysis. The results correlated well with strain identification by symptom phenotypes produced on differential cultivars inoculated with strains and isolates. The sensitivity of RT-PCR enabled detection of SMV from plants with necrotic symptoms in which the number of virus particles was too low to be detected by enzyme-linked immunosorbent assay.

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