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C S Bozarth

Publications and source records attributed to C S Bozarth.

4 recordsLinked to original sources

The turnip yellow mosaic virus tRNA-like structure cannot be replaced by generic tRNA-like elements or by heterologous 3' untranslated regions known to enhance mRNA expression and stability.

The tRNA-like structure (TLS) at the 3' end of the turnip yellow mosaic virus genome was replaced with heterologous tRNA-like elements, and with a poly(A) tail, in order to assess its role. Replacement with the valylatable TLSs from two closely related tymoviruses resulted in infectious viruses. In contrast, no systemic symptoms on plants, and only low viral accumulations in protoplasts, were observed for three chimeric genomes with 3' sequences known to enhance mRNA stability and translatability. One of these chimeras had a poly(A) tail, and the others had the TLS with associated upstream pseudoknot tracts from the 3' ends of brome mosaic and tobacco mosaic viruses. The latter two chimeric RNAs were shown to be appropriately folded by demonstrating their aminoacylation in vitro with tyrosine and histidine, respectively. The results show that enhancement of genome stability or gene expression is not the major role of the turnip yellow mosaic virus TLS. The major role is likely to be replicational, dependent on features present in tymoviral TLSs but not in generic tRNA-like structures.

Base Sequence↗

Expression of ORF-69 of turnip yellow mosaic virus is necessary for viral spread in plants.

Turnip yellow mosaic virus (TYMV) RNA has two extensively overlapping open reading frames (ORFs) encoding noncapsid proteins. The longest of these, ORF-206, is essential for RNA replication. We have investigated the expression and role of ORF-69 (encoding a protein with a MW of 69K) using specific antiserum and mutant genomic RNAs designed to interrupt ORF-69 expression. TYC69 antiserum immunoprecipitated a protein with an apparent MW of 75K (p69) from in vitro translations of TYMV RNA. Mutant RNAs with stop codons inserted at nucleotides 139, 178, and 178/224 in ORF-69 expressed ORF-69 at very low levels in vitro. These mutants replicated to wild-type levels in turnip or Chinese cabbage protoplasts, but were not recovered from any leaves of inoculated plants. These results suggest that ORF-69 products are dispensible for replication, but are required for viral spread. Revertant and pseudorevertant viruses with uninterrupted ORF-69 were recovered from plants showing delayed onset of symptoms after inoculation with two of the above mutants, indicating the importance of ORF-69 expression in establishing a systemic infection in plants. ORF-69 expression could be detected using Western blots in extracts from young, symptomatic leaves of infected plants. A single band with an apparent MW of 75-80K was detected in leaves infected with cDNA-derived viral RNAs, while a doublet was detected after infection with type strain viral RNA, suggesting the presence of two ORF-69 alleles or differential post-translational modification.

Blotting, Western↗

Cell wall proteins at low water potentials.

We investigated the proteins extractable from cell walls of stem tissues when plants were subjected to low water potentials (low psi(w)). Dark-grown soybean seedlings (Glycine max [L.] Merr.) showed decreased stem growth when the roots were exposed to vermiculite having low water content (psi(w) = -3 bar). After a time, growth resumed but at a reduced rate relative to the controls. The extractable protein increased in the cell walls as psi(w) decreased, especially a 28-kilodalton protein in the young tissue. In contrast, a 70 kilodalton protein, mainly extractable from mature cell walls, appeared to decrease slightly at low psi(w). No hydroxyproline was present in either protein, which shows that neither protein is related to extensin. The level of the 28 kilodalton protein increased in the cell wall of the dividing region soon after the initial growth inhibition, and it appeared in the elongating tissue at about the time growth resumed. The correlation between growth and these protein changes suggests that the two events could be related.

Journal Article↗

Photosynthetic Development of Anaerobically Grown Rice (Oryza sativa) after Exposure to Air.

During anaerobic germination, rice produces a coleoptile devoid of carotenoid and chlorophyll. Further development and greening of the shoot occur upon exposure of the seedlings to air. In this study, a comparison was made between anaerobically (N(2)) germinated rice, greened upon exposure to air, and air/dark (A/D) germinated seedlings, greened upon exposure to light. After exposure to air, N(2)-grown seedlings had a 76-hour lag before net oxygen evolution occurred compared to a 6-hour lag for A/D-grown seedlings. After 98 h of greening, N(2)-grown seedlings reached a rate of oxygen evolution equivalent to that of A/D-grown seedlings after 24 hours. Chlorophyll and carotenoid content showed a similar lag, but did not reach the level found in A/D-grown seedlings even after 124 hours of exposure to air. RuBPcase activity also lagged in N(2)-grown seedlings, ultimately reaching greater values than in the ;greened' A/D-grown seedlings. Phosphoenolpyruvate carboxylase activity was constant and low in all treatments except for a transient increase after 24 hours of greening of the N(2)-grown seedlings.

Journal Article↗