In Vitro Rearing of Perkinsiella saccharicida and the Use of Leaf Segments to Assay Fiji disease virus Transmission

Fiji leaf gall (FLG) is caused by the Reovirus, Fiji disease virus (FDV), which is transmitted to sugarcane by planthoppers of the genus Perkinsiella. Low vector transmission rates and slow disease symptom development make experimentation within the FDV-Perkinsiella-sugarcane system inherently difficult. A laboratory-based technique was devised to rear the vector using sugarcane leaves as a food source. Planthoppers were reared on sugarcane leaf segments embedded in agarose enclosed within plastic containers. To provide a nondestructive assay for determination of the inoculation potential of planthoppers, FDV was detected by reverse transcription-polymerase chain reaction (RT-PCR) in newly infected sugarcane leaf segments following exposure to viruliferous planthoppers. Leaf segment inoculation correlated with development of FLG symptoms in whole plants that were fed on by the same planthoppers. Analysis of FDV RNAs within the planthopper, measured by quantitative RT-PCR (qRT-PCR), indicated that FDV RNA concentration was associated with successful inoculation of the leaf segment, transmission of FDV to sugarcane and subsequent development of FLG in plants. Quantification of FDV RNA within planthoppers provided an additional measure to assess vector competence in individuals.

Improving Fiji disease resistance screening trials in sugarcane by considering virus transmission class and possible origin of Fiji disease virus

Fiji disease virus is a propagative, persistently transmitted virus that multiplies in species of the delphacid planthopper genus Perkinsiella, and in sugarcane, the feeding host of the insect. Efforts to improve and modify the disease rating system for Fiji disease have largely focussed on the planthopper as individual vectors of the virus, rather than as a population of the principal, or at least an alternative, host of the virus. This perspective has resulted in key parameters of disease incidence resulting from plant infection by propagative, persistently transmitted viruses being largely overlooked or misunderstood during efforts to improve the rating system. These parameters include the relatively long acquisition, latency, and transmission times, the percentage of the population containing virus, or viruliferous, in the above periods, and the effects of population density and number of plants visited on disease incidence. Suggestions to modify trial design to improve virus transmission to the plant, based on the disease incidence parameters of the propagative, persistent transmission class, are presented and the practical difficulties of implementing these proposals are discussed. In the context of fully understanding the underlying biology of this virus–insect–plant system, the hypothesis that Fiji disease virus, as a plant-infecting member of the Reoviridae, is primarily an insect virus with a secondary plant host, and may have diverged from an insect-infecting virus relatively recently is proposed and compared with other members of the family Reoviridae.

Production and evaluation of transgenic sugarcane containing a Fiji disease virus (FDV) genome segment S9-derived synthetic resistance gene

A transgenic line of the sugarcane cultivar Q124 with significantly enhanced resistance to Fiji disease was produced by microprojectile-mediated transformation with a transgene encoding a translatable version of Fiji disease virus (FDV) segment 9 ORF 1 under the control of the maize polyubiquitin promoter. Sixty-four transgenic lines were tested in glasshouse trials by caging the plants with viruliferous Perkinsiella saccharicida planthoppers. After 2 weeks, the planthoppers were removed and the plants monitored for symptoms. One transgenic line showed significantly enhanced resistance to Fiji disease compared with the Q124 parent and other lines showed varying levels of resistance. The molecular phenotypes of the transgenic plants at both the DNA and RNA levels were not entirely consistent with a resistance mechanism based on post-transcriptional gene silencing but were consistent with reports from other sugarcane-virus resistance systems. This is the first report of transgenic sugarcane containing an FDV-derived synthetic resistance gene showing resistance to FDV, although the mechanism of resistance has not yet been elucidated.

Fiji disease virus. [Distribution map].

A new distribution map is provided for Fiji disease virus Viruses: Reoviridae: Fijivirus Hosts: Sugarcane (Saccharum officinarum) and other Saccharum spp. Information is given on the geographical distribution in ASIA, Indonesia, Irian Jaya, Sulawesi, Malaysia, Peninsular Malaysia, Philippines, Thailand, AFRICA, Madagascar, OCEANIA, Australia, New South Wales, Queensland, Fiji, New Caledonia, Papua New Guinea, Samoa, Solomon Islands, Tonga, Vanuatu.