Citrus tristeza virus: characterization of Texas isolates, studies on aphid transmission and pathogen-derived control strategies

Citrus tristeza virus (CTV), an economically important graft-transmissible pathogen of citrus, causes major global declines in citrus production. In the commercial citrus of the Lower Rio Grande Valley of Texas (LRGV), where red grapefruit on tristeza-decline sensitive sour orange rootstocks predominate, incidence of CTV is low. The efficient CTV vector, the brown citrus aphid (BrCA, Toxoptera citricida Kirkaldy) is now established in Mexico and Florida, thus information is needed on the severity of CTV, CTV aphid transmission and the performance of transformed citrus towards CTV before T. citricida arrives in Texas so that appropriate management strategies can be selected. Biological indexing and molecular typing were performed on fifteen Texas CTV isolates. The majority of the CTV isolates tested contained the most severe CTV types known. In Florida, T. citricida were fed on crude CTV preparations in vitro and could transmit CTV to virus-free receptor plants with two CTV isolates, whereas a more highly purified CTV preparation from one CTV isolate was not transmitted by T. citricida. There were no differences in the majority of treatments in infectivity neutralizations using three CTV-derived antibodies (p25, p27 and p20). CTV p20 antibodies significantly enhanced the occurrence of CTV transmission in one test. The CTV genome of isolate H33 was sequenced using 'shot gun' methods. The H33 major component and H33 minor components were phylogenetically compared to the six other full-length CTV sequences. An untranslatable CTV coat protein gene was genetically transformed into the genome of the Texas commercial Rio Red grapefruit variety, and fifty-two independent transgenic lines were produced. CTV challenge responses by the transgenic lines were variable. Individual plants could be identified which had low virus titers by ELISA detection, a temporal decrease in virus titer, or a delay in virus titer accumulation. Comparing all wild types to all transgenic lines over every assessment revealed significant decreases in virus titer in the transgenic lines compared to that of the wild type. An RNA entity with similarities to marafiviruses was identified in a CTV infected plant. The entity appears non-graft transmissible to citrus, and non-mechanically transmissible to a range of herbaceous species.