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Pesquisa : G05.360.340.370.200 [Categoria DeCS]
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Id: biblio-1177370
Autor: Arévalo Gallegos, Sigifredo; Varela Rodríguez, Hugo; Lugo Aguilar, Héctor; Siqueiros Cendón, Tania S; Iglesias Figueroa, Blanca F; Espinoza Sánchez, Edward A; Aguado Santacruz, Gerardo A; Rascón Cruz, Quintín.
Título: Transient expression of a green fluorescent protein in tobacco and maize chloroplast
Fonte: Electron. j. biotechnol;45:1-9, May 15, 2020. ilus.
Idioma: en.
Resumo: BACKGROUND: Maize is one of the most important crops worldwide and has been a target of nuclear-based transformation biotechnology to improve it and satisfy the food demand of the ever-growing global population. However, the maize plastid transformation has not been accomplished due to the recalcitrant condition of the crop. RESULTS: In this study, we constructed two different vectors with homologous recombination sequences from maize (Zea mays var. LPC13) and grass (Bouteloua gracilis var. ex Steud) (pZmcpGFP and pBgcpGFP, respectively). Both vectors were designed to integrate into rrn23S/rrn16S from an inverted repeat region in the chloroplast genome. Moreover, the vector had the mgfp5 gene driven by Prrn, a leader sequence of the atpB gene and a terminator sequence from the rbcL gene. Also, constructs have an hph gene as a selection marker gene driven by Prrn, a leader sequence from rbcL gene and a terminator sequence from the rbcL gene. Explants of maize, tobacco and Escherichia coli cells were transformed with both vectors to evaluate the transitory expression­an exhibition of green and red fluorescent light under epifluorescence microscopy. These results showed that both vectors were expressed; the reporter gene in all three organisms confirmed the capacity of the vectors to express genes in the cell compartments. CONCLUSIONS: This paper is the first report of transient expression of GFP in maize embryos and offers new information for genetically improving recalcitrant crops; it also opens new possibilities for the improvement in maize chloroplast transformation with these vectors.
Descritores: Tabaco/metabolismo
Cloroplastos/genética
Cloroplastos/metabolismo
Zea mays/genética
Proteínas de Fluorescência Verde/metabolismo
-Transformação Genética
Biotecnologia
Reação em Cadeia da Polimerase
Plantas Geneticamente Modificadas
Plastídeos/genética
Proteínas de Fluorescência Verde/genética
Escherichia coli
Genoma de Cloroplastos
Responsável: CL1.1 - Biblioteca Central


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Id: biblio-1124206
Autor: Li, Bin; Lin, Furong; Huang, Ping; Guo, Wenying; Zheng, Yongqi.
Título: Development of nuclear SSR and chloroplast genome markers in diverse Liriodendron chinense germplasm based on low-coverage whole genome sequencing
Fonte: Biol. Res;53:21, 2020. tab, graf.
Idioma: en.
Projeto: National Natural Science Foundation of China; . National Forest and Grass Germplasm Resources Bank Program.
Resumo: BACKGROUND: Liriodendron chinense ranges widely in subtropical China and northern Vietnam; however, it inhabits several small, isolated populations and is now an endangered species due to its limited seed production. The objective of this study was to develop a set of nuclear SSR (simple sequence repeats) and multiple chloroplast genome markers for genetic studies in L. chinense and their characterization in diverse germplasm. RESULTS: We performed low-coverage whole genome sequencing of the L. chinense from four genotypes, assembled the chloroplast genome and identified nuclear SSR loci by searching in contigs for SSR motifs. Comparative analysis of the four chloroplast genomes of L. chinense revealed 45 SNPs, 17 indels, 49 polymorphic SSR loci, and five small inversions. Most chloroplast intraspecific polymorphisms were located in the interspaces of single-copy regions. In total, 6147 SSR markers were isolated from low-coverage whole genome sequences. The most common SSR motifs were dinucleotide (70.09%), followed by trinucleotide motifs (23.10%). The motif AG/TC (33.51%) was the most abundant, followed by TC/AG (25.53%). A set of 13 SSR primer combinations were tested for amplification and their ability to detect polymorphisms in a set of 109 L. chinense individuals, representing distinct varieties or germplasm. The number of alleles per locus ranged from 8 to 28 with an average of 21 alleles. The expected heterozygosity (He) varied from 0.19 to 0.93 and the observed heterozygosity (Ho) ranged from 0.11 to 0.79. CONCLUSIONS: The genetic resources characterized and tested in this study provide a valuable tool to detect polymorphisms in L. chinense for future genetic studies and breeding programs.
Descritores: Polimorfismo Genético/genética
Genoma de Planta/genética
Liriodendron/genética
Genoma de Cloroplastos/genética
-Primers do DNA/genética
DNA de Plantas/genética
Repetições de Microssatélites
Alelos
Sequenciamento Completo do Genoma
Genótipo
Responsável: CL1.1 - Biblioteca Central



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