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Characterization of the complete chloroplast genome of Rohdea wattii (Asparagaceae).

ABSTRACT: As a kind of rare plant with important medicinal value and breeding value, Rohdea wattii usually grow in dark and damp places. In this study, we reported the complete chloroplast genome of R. wattii. We used a variety of bioinformatics analysis methods to analyze and visualize the chloroplast genome and phylogenetic relationships. A typical quadripartite structure was observed in the chloroplast genome of R. wattii with a genome size of 156,487 bp. The length of the large single-copy region (LSC) was 84,969 bp, which was the longest region among the four regions; the second longest was the Inverted repeat region (IR) with 26,451 bp in length; and the smallest was the small single-copy region (SSC) with 18,616 bp in length. The overall GC content of R. wattii chloroplast genome is 37.61%. In addition, a total of 132 genes were identified in the whole genome of R. wattii, which include 86 protein-coding genes, 38 tRNA genes, and eight rRNA genes. Finally, by constructing a phylogenetic tree to analyze the phylogeny of R. wattii, and it indicated that R. wattii and Rohdea chinensis are a close evolutionary relationship.

SUBMITTER: Wang Y

PROVIDER: S-EPMC8405109 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Erigeron breviscapus is a famous medicinal plant. However, the limited chloroplast genome information of E. breviscapus, especially for the chloroplast DNA sequence resources, has hindered the study of E. breviscapus chloroplast genome transformation. Here, the complete chloroplast (cp) genome of E. breviscapus was reported. This genome was 152,164bp in length, included 37.2% GC content and was structurally arranged into two 24,699bp inverted repeats (IRs) and two single-copy areas. The sizes of the large single-copy region and the small single-copy region were 84,657 and 18,109bp, respectively. The E. breviscapus cp genome consisted of 127 coding genes, including 83 protein coding genes, 36 transfer RNA (tRNA) genes, and eight ribosomal RNA (rRNA) genes. For those genes, 95 genes were single copy genes and 16 genes were duplicated in two inverted regions with seven tRNAs, four rRNAs, and five protein coding genes. Then, genomic DNA of E. breviscapus was used as a template, and the endogenous 5' and 3' flanking sequences of the trnI gene and trnA gene were selected as homologous recombinant fragments in vector construction and cloned through PCR. The endogenous 5' flanking sequences of the psbA gene and rrn16S gene, the endogenous 3' flanking sequences of the psbA gene, rbcL gene, and rps16 gene and one sequence element from the psbN-psbH chloroplast operon were cloned, and certain chloroplast regulatory elements were identified. Two homologous recombination fragments and all of these elements were constructed into the cloning vector pBluescript SK (+) to yield a series of chloroplast expression vectors, which harbored the reporter gene EGFP and the selectable marker aadA gene. After identification, the chloroplast expression vectors were transformed into Escherichia coli and the function of predicted regulatory elements was confirmed by a spectinomycin resistance test and fluorescence intensity measurement. The results indicated that aadA gene and EGFP gene were efficiently expressed under the regulation of predicted regulatory elements and the chloroplast expression vector had been successfully constructed, thereby providing a solid foundation for establishing subsequent E. breviscapus chloroplast transformation system and genetic improvement of E. breviscapus.

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Characterization of the complete chloroplast genome of Rohdea wattii (Asparagaceae).

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