Citrus tristeza virus (CTV) exists in field isolates as a complex of virus isolates. This complex may contain both mild and severe CTV. Using single and multiple aphid transmissions, subiso-lates of the various field isolates were separated. Some CTV isolates that tested negative with the monoclonal antibody MCA13 consisted of MCA13-positive subisolates. Using primers to specific and variable regions of the CTV genome, molecular profiles of the isolates and subisolates were generated and compared. The profiles of the subisolates sometimes were very different from the parent field isolates from which they were transmitted.
Some strains of the Citrus tristeza virus (CTV) cause stem pitting in sweet orange (Citrus sinensis (L.) Osbeck). This abnormality causes tree decline and reduction in fruit size and yield of affected citrus trees. Stem-pitting symptoms can occur on trunks, on all sizes of limbs, and on the twigs where fruit are produced. Variously sized pits or grooves in the wood often contain a yellow gum. Irregular growth of the phloem occurs in the area of these xylem pits. The histology of stem pitting caused by an Australian CTV isolate was studied in sweet orange using light and electron microscopy. Using scanning electron microscopy, details of the wood pits containing the gumming material were revealed. In thin sections of bark tissue, outgrowths of the phloem tissue were found at various intervals that corresponded to the pits in the wood. Higher numbers of viral inclusions were detected in the phloem outgrowths than were present in the other sieve elements.
Greasy spot, caused by Mycosphaerella citri, is a serious disease of citrus in the Caribbean basin. M. citri is a loculoascomycete and produces pseudothecia in decomposing leaves after intermittent wetting and drying. A new in vitro mating technique was developed for production of pseudothecia on sterilized leaf disks in petri dishes. Of the single-ascospore cultures that were recovered from individual asci, four were one mating type and four were a second mating type (tentatively designated mat+ and mat-), indicating that M. citri probably is heterothallic and bipolar like most other loculoascomycetes. Most populations of ascospores recovered from individual leaves or from leaves from groves of different citrus species and various locations had a 1:1 ratio of mating types consistent with random mating. Cytological studies demonstrated that the ontogeny of pseudothecial development was similar to other loculoascomycetes. The formation of mature pseudothecia required 30 to 45 cycles of wetting and drying of infected, dead leaves which required approximately 60 to 90 days. The in vitro system for pseudothecial production and the knowledge of the mating system in M. citri will facilitate genetic studies of this important pathogen.
Citrus chlorotic dwarf (CCD) is a virus-like disease discovered in Turkey in the mid 1980s. Infected plant material was sent to the USDA, ARS exotic citrus quarantine facility in Beltsville, MD and was grafted into Citrus macrophylla seedlings. Symptoms of CCD include various chlorotic patterns and leaf crinkling or other types of distortion. Leaves are small and often have a notch near the apex. Leaves showing symptoms of chlorosis, leaf crinkling and distortion, and bark from the symptomatic shoots were collected and prepared for ultrathin sectioning and transmission electron microscopy. In sections of tissue, high numbers of ribosomes and endoplasmic reticulum (ER), especially rough ER, were observed in both leaf and bark tissues. Chloroplasts showed partial to complete breakdown of the grana in all tissues and very little starch was observed. Cellular membranes and cell walls remained intact and most cell contents were recognizable. In bark sections, there were numerous membrane-bound vesicles and some phloem cells were found to contain an unknown filamentous material, which appeared virus-like.
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