Author: Tromas, Nicolas; Zwart, Mark P.; Forment, Javier; Elena, Santiago F.
Title: Shrinkage of Genome Size in a Plant RNA Virus upon Transfer of an Essential Viral Gene into the Host Genome Document date: 2014_2_20
ID: 5fejitls_12
Snippet: Two evolution experiments were performed with these viruses. In the first experiment, conceived to study the appearance of genomic deletions de novo, four 3-week passages of TEV and TEV-ÃNIb were performed in N. tabacum 35S::NIb plants. At 21 dpi, all symptomatic leaves of each of the five lineages per virus were collected. Five hundred milligrams of symptomatic tissue was ground in 500 ml of grinding buffer (50 mM potassium phosphate pH ¼ 7.0,.....
Document: Two evolution experiments were performed with these viruses. In the first experiment, conceived to study the appearance of genomic deletions de novo, four 3-week passages of TEV and TEV-ÃNIb were performed in N. tabacum 35S::NIb plants. At 21 dpi, all symptomatic leaves of each of the five lineages per virus were collected. Five hundred milligrams of symptomatic tissue was ground in 500 ml of grinding buffer (50 mM potassium phosphate pH ¼ 7.0, 3% polyethyleneglycol 6000). Ten microliters of the resulting sap was then mixed with inoculation buffer and used to infect a batch of five healthy transgenic plants by abrasion for each TEV genotype. In the second experiment, conceived to evaluate the subsequent evolution and fitness optimization of genotypes missing the NIb cistron, 30 one-week passages of TEV and TEV-ÃNIb were performed in N. tabacum 35S::NIb plants. At 7 dpi, symptomatic leaves were collected for each of the five lineages per virus. Infectious sap was prepared as described earlier.
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