RESUMO
Rhodophiala bifida (R. bifida) is a representative of the Amaryllidaceae plant family and is rich in montanine, an alkaloid with high pharmaceutical potential. Despite the interest in these compounds, many steps of the biosynthetic pathway have not been elucidated. In this study, we identified the alkaloids produced in different organs of R. bifida under different growth conditions, set up the conditions for in vitro R. bifida regeneration and initiated the molecular characterization of two R. bifida genes involved in alkaloids biosynthesis: the Norbelladine 4'-O-Methyltransferase (RbN4OMT) and the Cytochrome P450 (RbCYP96T). We show that montanine is the main alkaloid produced in the different R. bifida organs and developed a direct organogenesis regeneration protocol, using twin-scale explants cultivated on media enriched with naphthalene acetic acid and benzyladenine. Finally, we analyzed the RbN4OMT and RbCYP96T gene expressions in different organs and culture conditions and compared them to alkaloid production. In different organs of R. bifida young, adult and regenerated plants, as well as under various growing conditions, the transcripts accumulation was correlated with the production of alkaloids. This work provides new tools to improve the production of this important pharmaceutical compound and for future biotechnological studies.
Assuntos
Alcaloides de Amaryllidaceae/metabolismo , Amaryllidaceae/metabolismo , Isoquinolinas/metabolismo , Amaryllidaceae/genética , Vias Biossintéticas , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Isoquinolinas/químicaRESUMO
The tribe Leucocoryneae is taxonomically and cytogenetically complex, mainly due to its extraordinary morphological and karyological variation. Robertsonian translocations had long been recognized as a central factor contributing to karyotype diversity within the Leucocoryneae, but so far no major tendency prevailing on the observed complexity of karyotype formula among species has been identified. The assessment of nuclear DNA contents by flow cytometry using propidium iodide in 23 species, representing all genera within the tribe, showed a monoploid genome size variation of 1Cx = 9.07-30.46 pg denoting a threefolds fluctuation. A highly significant linear association between the average DNA content per chromosome arm (2C/FN) and the monoploid genome size (1Cx) is reported for the first time and identified as a novel indicator of a trend governing karyotype diversity within Leucocoryneae. This trend shows that a reduction in DNA content per chromosome arm is influencing and has shaped karyotype evolution of different monophyletic groups within the tribe despite the complex karyotype diversity and apparently contrasting patterns of genome sizes.
Assuntos
Amaryllidaceae/genética , Evolução Molecular , Tamanho do Genoma , Genoma de Planta , Cariótipo , Citometria de Fluxo , PropídioRESUMO
Agapanthus (Agapanthaceae) has 10 species described. However, most taxonomists differ respect to this number because the great phenotypic plasticity of the species. The cytogenetic has been an important tool to aid the plant taxon identification, and to date, all taxa of Agapanthus L'Héritier studied cytologically, presented 2n = 30. Although the species possess large chromosomes, the group is karyologically little explored. This work aimed to increase the cytogenetic knowledge of Agapanthus africanus (L.) Hoffmanns by utilization of chromosome banding techniques with DAPI / CMA3 and Fluorescent in situ Hybridization (FISH). In addition, flow cytometry was used for determination of DNA content and the percentage of AT / GC nitrogenous bases. Plants studied showed 2n = 30 chromosomes, ranging from 4.34 - 8.55 µm, with the karyotype formulae (KF) = 10m + 5sm. Through FISH, one 45S rDNA signal was observed proximally to centromere of the chromosome 7, while for 5S rDNA sites we observed one signal proximally to centromere of chromosome 9. The 2C DNA content estimated for the species was 2C = 24.4 with 59% of AT and 41% of GC. Our data allowed important upgrade for biology and cytotaxonomy of Agapanthus africanus (L.) Hoffmanns.