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求大神帮忙翻译三段话
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Seed Germination and Seedling Establishment Assays After surface sterilization of the seeds, stratification was conducted in the dark at 4°C for 3 d. Next, ;100 seeds of each genotype were sown on MS plates lacking or supplemented with 0.5 mM ABA, 150 mM NaCl, or 400 mM mannitol. In the analyses of iDDA1 lines, 1 mMABAwas used and b-estradiol was added to medium at 10 mM final concentration as stated. To score seed germination, radicle emergence was analyzed at 72 and 96 h after sowing. Seedling establishment was scored after 5 d as the percentage of seeds that developed green expanded cotyledons and the first pair of true leaves. Root Growth Assays Seedlings were grown on vertically oriented MS plates for 4 to 5 d. Afterward, 20 plants were transferred to new MS plates lacking or supplemented with 10 mM ABA. The plates were scanned on a flatbed scanner after 10 d to produce image files suitable for quantitative analysis of root growth using ImageJ version 1.37 software Phylogenetic Analysis DDA1 homologs were searched in different databases, Plaza (https:// bioinformatics.psb.ugent.be/plaza/), Phytozome (https://www.phytozome.net), Gramene (https://www.gramene.org), TAIR (https://www.arabidopsis.org), and GenBank (https://www.ncbi.nlm.nih.gov), using web-hosted BLAST applications (https://blast.ncbi.nlm.nih.gov/Blast.cgi). All truncated DDA1 sequences were excluded in subsequent analyses. Sequences were aligned using ClustalW2 (https://www.ebi.ac.uk/Tools/msa/clustalw2/) with the Gonnet protein comparison matrix and default parameters (gap opening penalty of 10 and gap extension penalty of 0.1 for initial pairwise alignment; gap opening penalty of 10 and gap extension penalty of 0.2 for the multiple alignment; with residue-specific and hydrophilic residue penalties, a gap separation distance of 4 and a 30% delay divergent cutoff) and then manually inspected to check for possible misalignments (Supplemental Data Set 1). The best model fitting the alignment was selected using ProtTest (Abascal et al., 2005), which corresponded to a Jones-Taylor- Thornton model for amino acid substitution with a discrete gamma distribution (four categories), a proportion of invariant, and an initial BIONJ tree. This model was used to conduct a maximum likelihood analysis with bootstrapping using 1000 replicates using the MEGA 5 (https://www. megasoftware.net/) software. The gaps/missing treatment parameter was set to complete deletion. The more distant homologs, from Selaginella moellendorffii and Picea glauca, were used as an outgroup to root the resulting phylogenetic tree. |
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玉砌的强: 金币+15, 翻译EPI+1, ★★★很有帮助 2014-12-09 21:54:10
玉砌的强: 金币+15, 翻译EPI+1, ★★★很有帮助 2014-12-09 21:54:10