The complete nucleotide sequence of an Albanian isolate of grapevine leafroll-associated virus 7 (GLRaV-7-Alb) was determined. The viral genome consists of 16,404 nucleotides and has nine open reading frames (ORFs) that potentially encode proteins, most of which are typical for members of the family Closteroviridae. Only the 25-kDa (ORF8) and 27-kDa (ORF9) proteins had no apparent similarity to other viral proteins in the sequence databases. The genome structure of GLRaV-7-Alb closely resembles that of little cherry virus 1 and cordyline virus 1. In phylogenetic trees constructed with HSP70h sequences, these three viruses cluster together in a clade next to that comprising members of the genus Crinivirus, to which they are more closely related than to the clostero- and ampeloviruses. The molecular properties of these three viruses differ sufficiently from those of members of the three extant genera of the family Closteroviridae to warrant their classification in a novel genus.
Grapevine leafroll-associated virus-7 (GLRaV-7) was transmitted from an Albanian grapevine accession to Tetragonia expansa by the parasitic dodder Cuscuta reflexa and to Nicotiana occidentalis by Cuscuta europea. Cuscuta campestris was infected by GLRaV-7 but could not transfer the virus to an experimental host. Transmission of the virus was verified by reverse transcription–polymerase chain reaction (RT-PCR) from total nucleic acid (TNA) and double-stranded RNA (dsRNA) extracts from all five plant species. DsRNA extractions separated on agarose gels showed strong visible bands corresponding to high-molecular-weight virus genome and to subgenomic RNA. GLRaV-7 was maintained in C. reflexa, C. campestris, T. expansa, and N. occidentalis for more than 4 years. Infected T. expansa and the Cuscuta species remained symptomless while N. occidentalis showed severe symptoms leading to stunting and decline of the plants. Quantitative PCR showed great differences in the titer of GLRaV-7 between the tissues of its natural and experimental host plants. This is the first report on a virus of the Closteroviridae that was successfully transmitted to an herbaceous plant by dodder. Virus replication could be demonstrated in Cuscuta. Both the new experimental hosts of GLRaV-7 and Cuscuta allowed extraction of dsRNA for further characterization of the viral genome, which previously required grapevine scraping of phloem. This is time-consuming and does not always lead to satisfactory results. These alternative hosts of GLRaV-7 facilitate nucleic acid extractions and could be used as model plants for etiological studies.
Size-related variation in physiological parameters as diverse as photosynthetic capacity, abscisic acid relationships or the relative water deficit at stomatal closure have been reported for a large number of vascular epiphytes, but the proximate mechanism behind these observations has not been identified. We test four possible reasons for size-related changes in photosynthetic capacity, leaf-N content and specific leaf area: (i) plant size itself, (ii) plant age or developmental stage, (iii) previous nutrition, or (iv) previous water regime. A suite of study species and approaches were used: a 'natural experiment' with the orchid Polystachya foliosa; an experimental field study with another orchid, Dimerandra emarginata; and a study under controlled conditions with the tank bromeliad, Vriesea sanguinolenta. Neither size, age nor differences in water supply caused differences in leaf N and photosynthetic capacity, while low supply of nutrients yielded, and high supply with nutrients completely removed, size-related trends. The observed size-related trends are thus a consequence of in situ differences in nutrient acquisition. Arguably, the improved nutrient status of larger plants under natural conditions results from larger tanks, holding moisture for increasingly longer intervals, which allows longer periods of decomposition of detritus and of nutrient uptake.
Gas exchange of the understorey epiphyte Aspasia principissa was studied in fluctuating light conditions both in the laboratory and in the field, testing the hypothesis that vascular epiphytes differ from most terrestrial understorey plants in showing a higher priority for water conservation. Consequently, a slow response of stomatal conductance to sudden increases in incident photon flux density (PFD) was expected, as was a fast loss of induction after such a light fleck. Results were only partly consistent with these expectations. Full induction of photosynthesis was indeed very slow and was not reached before, respectively, 40 and 60 min of saturating PFD in the field and the laboratory. In contrast, kinetics of induction loss were comparable with those of most terrestrial species studied to date. The overall impact of light flecks on in situ carbon gain again fulfilled expectations, being rather limited: the observed carbon gain was only approx. 66% of the potential carbon gain estimated from a square-wave response model. It is concluded that in the drought-prone epiphytic habitat of a moist lowland forest, water conservation takes priority over carbon gain, which severely limits the use of light flecks for CO(2) fixation in vascular epiphytes.
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