Others are held in NCBI database with accession number KX595275.įunding: The project was funded by the Natural Science Foundation of Shanghai (18ZR1417400) (CC), the Shanghai Port and Offshore Ecological Environment Technology Service Platform Project (19DZ2292500) (CC), the Key Laboratory of Integrated Marine Monitoring and Applied Technologies for Harmful Algal Blooms, S.O.A. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.ĭata Availability: The mainly relevant data are contained within the paper and its Supporting information files. Received: AugAccepted: ApPublished: June 1, 2021Ĭopyright: © 2021 Cai et al. PLoS ONE 16(6):Įditor: Berthold Heinze, Austrian Federal Research Centre for Forests BFW, AUSTRIA compressa, the findings of this study have implications for the rapid non-sequencing detection of this species during the occurrence of green tides in the region.Ĭitation: Cai C, Gu K, Zhao H, Steinhagen S, He P, Wichard T (2021) Screening and verification of extranuclear genetic markers in green tide algae from the Yellow Sea. Owing to the high morphological plasticity of U. ![]() mutabilis but not in other algal species of the Yellow Sea. ![]() Finally, the tandem repetition was identified in the mitogenome of U. Interestingly, a few tandem repetitions were significant for some Ulva subspecies and relatively more evident in mitochondria than in chloroplasts. Complementary/forward/palindromic interval repetitions were more abundant in chloroplast genomes than in mitogenomes. From phylogenetic analysis, we identified the petA chloroplast genes as potential genetic markers that are similar to the tufA marker. In addition, phylogenetic analyses of the aligned genes from their chloroplast genomes and mitogenomes confirmed the evolutionary trends of the extranuclear genomes. ohnoi were more divergent, especially in sharing complementary/palindromic repetitions. Chloroplast genomes were clustered together in the entire phylogenetic tree and shared several forward/palindromic/tandem repetitions, similar to those in U. mutabilis chloroplast genomes had similar gene queues, with individual genes exhibiting high homology levels. ![]() The generated phylogenetic tree was analyzed based on single and aligned genes in the chloroplast genome of Ulva compared to mitogenome genes to detect evolutionary trends. We compared the chloroplast genome with the mitogenome. compressa (alpha-numeric code: RD9023) and focused on the assessment of genome length, homology, gene order and direction, intron size, selection strength, and substitution rate. In the present study, we sequenced and annotated the complete chloroplast genome of U. Therefore, GFP could be used as a vital marker to improve the transformation efficiency and to easily monitor the segregation of marker genes, thus facilitating screening of marker-free progeny.Over the past decade, Ulva compressa, a cosmopolitan green algal species, has been identified as a component of green tides in the Yellow Sea, China. Co-expression of mgfp5 and nptII was found in 28 out of 29 T1 lines, and segregation of the reporter beta-glucuronidase gene, gusA, from mgfp5 to nptII was found in 12 out of 29 T1 lines. The frequency of co-transformants produced by this vector was 56.3%. A dual marker double T-DNA vector was then constructed by assembling the green fluorescent protein (GFP) gene mgfp5 and the neomycin phosphotransferase gene nptII into the same T-DNA. Co-transformation methods using a double T-DNA vector or using mixture of two Agrobacterium tumefaciens strains were compared, and showed that the double T-DNA vector method could produce marker-free transgenic tobacco (Nicotiana tabacum L.) plants more efficiently. We investigated the potential of a novel double T-DNA vector for generating marker-free transgenic plants.
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