First and third authors: International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Patancheru 502 324, India; and second author: Scottish Crop Research Institute, Invergowrie DD2 5DA, Scotland, U.K.
ABSTRACT
The agent of sterility mosaic, a disease that is a major constraint on pigeonpea (Cajanus cajan) production in the Indian subcontinent, is transmitted by the eriophyid mite, Aceria cajani. This agent has remained elusive for decades despite intensive efforts but we report the isolation of highly flexuous filamentous virus-like particles (VLPs) of 3 to 10 nm in width and of undefined lengths from sterility mosaic disease (SMD)-affected pigeonpea plants. Purified VLP preparations from virus-infected pigeonpea and Nicotiana benthamiana had a buoyant density in cesium chloride of 1.22 to 1.23 g cm-3 and contained a major virus-specific protein species of ≈32 kDa and 5 to 7 RNA species of ≈6.8 to 1.1 kb. The sequence of some complementary DNA clones to RNA from purified VLP preparations had no significant matches in database searches. Two oligonucleotide primers derived from one such sequence, when used in reverse transcriptase-polymerase chain reaction assays, amplified a product of 321 bp specifically from SMD-affected pigeonpea plants. Purified VLP preparations were used to produce polyclonal antibodies that, in infected plants, detected the virus using enzyme-linked immuno-sorbent assay (ELISA) and the virus-specific 32-kDa protein in western immunoblotting (WIB). In such assays, the virus was detected consistently in all SMD-affected pigeonpea plant samples from several different locations in India, but not in samples from symptom-free pigeonpea plants from the same locations. In experimental studies, all pigeonpea plants inoculated with viruliferous A. cajani and those plants graft-inoculated with SMD-affected tissue were infected with the virus as assessed by ELISA and WIB, but not any uninfected pigeonpea plants. This virus, tentatively named Pigeonpea sterility mosaic virus (PPSMV), has some properties similar to virus species in the genera Tospovirus and Tenuivirus and with the eriophyid mite-transmitted High plains virus (HPV) but is distinct from these and from all other characterized viruses. The combination of novel properties shown by PPSMV and HPV suggest that they may constitute species in a new genus of plant viruses.
