ABSTRACT
PURPOSE: Poncirus trifoliata has been reported to have anti-inflammatory, antioxidant, and immune activities. However, its anti-obesity activity and the mechanism by which the water extract of dried, immature fruit of Poncirus trifoliata (PF-W) acts are not clear. This study suggests a potential mechanism associated with the anti-obesity activity of PF-W. METHODS: We measured the effect of PF-W on lipoprotein lipase (LPL) regulation using enzyme-linked immunosorbent assay (ELISA) and an activity assay. The LPL regulation mechanism was examined by reverse transcription polymerase chain reaction (RT-PCR) to measure the mRNA expression of biomarkers related to protein transport and by western blot for analysis of the protein expression of the transcription factor CCAAT-enhancer-binding protein (C/EBPbeta) RESULTS: The total polyphenol and flavonoid content of PF-W was 52.15 +/- 4.02 and 6.56 +/- 0.47 mg/g, respectively. PF-W treatment decreased LPL content in media to 58 +/- 5% of that in control adipocyte media, and increased LPL content to 117 +/- 3.5% of that in control adipocytes, but did not affect the mRNA expression of LPL. PF-W also increased the mRNA expression of sortilin-related receptor (SorLA), a receptor that induces endocytosis and intracellular trafficking of LPL, in a concentration- and time-dependent manner. Finally, cell fractionation revealed that PF-W treatment induced the expression of C/EBPbeta, a SorLA transcription factor, in the nuclei of 3T3-L1 adipocytes. CONCLUSION: The LPL secretion and activity assay showed PF-W to be an LPL secretion inhibitor, and these results suggest the potential mechanism of PF-W involving inhibition of LPL secretion through C/EBPbeta-mediated induction of SorLA expression.
Subject(s)
Adipocytes , Biomarkers , Blotting, Western , CCAAT-Enhancer-Binding Proteins , Cell Fractionation , Endocytosis , Enzyme-Linked Immunosorbent Assay , Fruit , Lipoprotein Lipase , Polymerase Chain Reaction , Poncirus , Protein Transport , Reverse Transcription , RNA, Messenger , Transcription Factors , WaterABSTRACT
Avaliou-se o desenvolvimento vegetativo inicial de porta-enxertos cítricos cultivados em casa de vegetação, por meio de semeadura e de repicagem de raiz nua. O experimento foi conduzido em casa de vegetação, na Estação Experimental Agronômica da UFRGS, em Eldorado do Sul, RS, a partir do mês de setembro de 2003, afim de testar a viabilidade de aproveitamento de plântulas oriundas do desbaste. O delineamento experimental adotado foi o de blocos casualizados, com quatro repetições, em esquema de parcela subdividida e cada subparcela constituída por 22 tubetes. Nas parcelas principais foram testados dois sistemas de propagação: semeadura e repicagem no momento do desbaste. Nas subparcelas foram testados três porta-enxertos cítricos: Trifoliata (Poncirus trifoliata [L.] Raf.), citrangeiro "C37" (P. trifoliata x Citrus sinensis [L.] Osbeck.) e limoeiro "Cravo" (C. limonia Osbeck.). A semeadura foi realizada em tubetes cônicos de polietileno preto (capacidade de 120cm³), com duas a três sementes por tubete. A repicagem foi feita no momento do desbaste da bandeja semeada quando as plântulas estavam, em média, com duas a quatro folhas. Os resultados demonstraram que há viabilidade de aproveitamento das plântulas resultantes do desbaste, sem que ocorra diferenças no desenvolvimento vegetativo. Além disso, foi observado que o porta-enxerto "C37" apresentou um vigor inicial maior que o Trifoliata e o limoeiro "Cravo".
The initial vegetative growth of citrus seedling rootstocks was evaluated by means of sowing and transplantation of bare-root seedling. The experiment was conduced in September 2003 in greenhouse conditions in the Estação Experimental Agronômica/UFRGS in Eldorado do Sul, Brazil. The objective of this study was to test the viability of use of seedlings deriving from thinning. The experiment was in randomized blocks with four replications in a split plot design and each sub-plot consisted of 22 tubs. In the main plot were tested two propagation systems: sowing and transplantation at the thinning moment. In sub-plots were tested three citrus rootstocks: Trifoliate orange (Poncirus trifoliata [ L. ] Raf.); 'C37' citrange (P. trifoliata x Citrus sinensis [ L. ] Osbeck.), and 'Rangpur' lime (C. limonia Osbeck.). The sowing was carried in conical tubs of black polyethylene (capacity of 120cm³) with two to three seeds in each tub. Transplantation was made at the thinning moment, when the seedlings had, in average, two to four leaves. The results demonstrate that it is possible to use seedlings resulting from thinning, without differences in the vegetative development comparatively to sowing. The 'C37' citrus rootstock presents a larger initial vigor when compared to the other citrus rootstocks tested.
ABSTRACT
Nearly 65,000 citrus EST (Expressed Sequence Tags) have been investigated using the CitEST project database. Microsatellites were investigated in the unigene sequences from Citrus spp. and Poncirus trifoliata. From these sequences, approximately 35 percent of the non-redundant ESTs contained SSRs. The frequencies of different SSR motifs were similar between Citrus spp and trifoliate orange. In general, mononucleotide repeats appeared to be the most abundant SSRs in the CitEST database, but we also identify di-, tri-, tetra-, penta- and hexanucleotide repeats. The AG/CT and AAG/CTT were the most common dinucleotide and trinucleotide motifs, with frequencies of 54.4 percent and 25.2 percent, respectively. Primer sequences flanking SSR motifs were successfully designed and synthesized. After in silico polymorphism analysis, a subset of sixty-eight primers was validated in different Citrus spp. and Poncirus trifoliata. PCR-amplification revealed polymorphism in citrus with all tested primer pairs and showed the potential of these markers for linkage mapping. Our study showed that the CitEST database can be exploited for the development of SSR markers that can amplify Citrus spp. and related genus for comparative mapping and other genetic analyses.
ABSTRACT
Object To establish an effective system for Agrobacterium mediated genetic transformation of Poncirus trifoliata Raf Methods The explants used for transformation were the epicotyls from P trifoliata; the Agrobacterium tumefaciens strain was EHA101, provided with the vector plasmid pGA482GG; the coat protein gene (CTV cp gene) was introduced into the transformation plasmid The transformation was proved by PCR and histochemical GUS assay Results In trans for mation of P tri foliata, 20 days epicotyls were suitable for transformation Shoot regeneration frequency was high when cocultivation time was 2-3 days The presence of acetosyringone during cocultivation could enhance the efficiency of transformation Histochemical GUS assay showed that 70 0% of the resistant plants were GUS positive Extra gene was proved to be transformed into P trifoliata plant by PCR analysis Conclusion An effective genetic transformation system has been established for P trifoliata This system will be useful for resistant breeding of P trifoliata