Molecular Characterization of Rhynchosia mosaic virus-Puerto Rico Associated with Symptomatic Rhynchosia minima and Cajanus cajan in Puerto Rico.

A begomovirus (family Geminiviridae) has long been suspected to be associated with Rhynchosia mosaic (RhM) disease of Rhynchosia minima (L.) DC., a weed that is widespread in Puerto Rico (PR). The suspect virus has been transmitted by the Sida biotype of Bemisia tabaci (Genn.) and has been designated RhM virus-PR (RhMV-PR) (1) (synonym, Rhynchosia mosaic virus [RMV]). RhM symptoms in R. minima included yellow foliar mosaic and stunting. The virus has a broad experimental host range and infects species in the Fabaceae, including R. minima, pigeon pea (Cajanus cajan (L.) Millsp.), and Clitoria falcata L. (1). However, until now RhMV has not been identified from naturally infected pigeon pea or Clitoria falcata. R. minima and C. cajan plants exhibiting yellow foliar mosaic and stunting symptoms were collected in Puerto Rico. Using the B biotype of B. tabaci as the vector, their whitefly transmissibility from the respective source plant to R. minima and C. cajan test plants was confirmed, and symptoms in inoculated host were indistinguishable for both isolates. Using polymerase chain reaction (PCR) and primers (2), three amplicons were obtained and cloned for each isolate. PCR products (1.1 and 2.1 kbp) were assembled (200 nucleotide [nt] overlap) to yield an apparent full-length DNA A component (2.6 kbp) containing the diagnostically informative viral coat protein gene (CP) and common region (CR-A). PCR primers were used to amplify the DNA B component segment (0.7 kbp) containing the CR-B (2). The DNA sequence for the core CP (533 nt) and full CP (750 nt) were compared with analogous sequences for well-studied begomoviruses, and CR-A and CR-B (153 nt) were compared for RhMV isolates. All isolates noted were obtained from GenBank. The core CP for isolates from R. minima (AF442117) and C. cajan (AY062025) shared 97.9% nucleotide identity (100% AA similarity) and the CR-A (AF442118) and CR-B (AF442119) sequences for R. minima and C. cajan isolates were ~96% identical, indicating the A and B components are of the same begomovirus. Comparison of the core CP sequence for an independent isolate from C. cajan from PR (AY028308) (4) with those for R. minima and C. cajan isolates indicated 95.5% (99.4% AA) and 96.2% (99.4% AA) nucleotide identity, respectively, indicating association of RhMV with both C. cajan samples. The recently archived core CP (533 nt) (AY028308) is actually of RhMV-PR, rather than a distinct begomovirus species, as indicated (4). Interestingly, the core CP of R. minima (AF442117) and C. cajan (AY062025) isolates were 91.7% (98.9% AA) and 92.3% (98.9% AA) identical, respectively, with a PR isolate from Clitoria falcata (AF070924), also confirming that RhMV-PR naturally infects Clitoria falcata. Analysis of the full CP for the R. minima and C. cajan isolates revealed that their closest relatives were Macroptilium mosaic virus (MaMV-PR) (AF176092) and Bean golden mosaic virus (BGMV-PR) (M10070) at 89 and 84% nucleotide identity, respectively. Applying the 90% CP rule (3) to RhMV CP sequences, RhMV is a distinct begomovirus species. At least three begomoviruses, BGMV-PR, MaMV-PR, and RhMV-PR, naturally infect leguminous species in Puerto Rico. References: (1) J. Bird. Phytopathology 52:286, 1962. (2) A. M. Idris and J. K. Brown. Phytopathology 88:648, 1998. (3) M. A. Mayo and C. R. Pringle. J. Gen. Virol. 79:649, 1998. (4) R. L. Rodriguez et al. Plant Dis. 85:1119, 2001.