RESUMO
BACKGROUND: Since its introduction to the Anglophone Caribbean in 1793, breadfruit has had a diverse history in the region, and there is a considerable repository of traditional knowledge about the crop, that is undocumented. Consequently, it remains underutilized as a food source, despite recognition of its potential to contribute to food and nutrition security. Understanding the folk taxonomy and traditional knowledge associated with its diversity and uses is a prerequisite to develop programs for its commercial production and utilization. METHOD: This study was conducted among 170 respondents who were selected across four Anglophone Caribbean countries and provided information on the ethnobotany and traditional knowledge associated with breadfruit biodiversity, including systems of naming, identification and classification of breadfruit cultivars or types. RESULTS: Breadfruit has socio-cultural and economic value and is produced for both home use and sale by most respondents (68%). The genetic diversity of breadfruit managed by the respondents is also important, as a total of 51 vernacular names were identified, with nine of those names recorded for the first time in this study. Breadfruit types were identified by morphological and agronomical characteristics, with other important traits relating to use and cooking quality. Classification of breadfruit cultivars or types was based on eating-quality, most suitable methods of preparation and ease of cooking. CONCLUSION: The ethnobotanical and traditional knowledge obtained from this study may be useful in assessing the genetic diversity of breadfruit and guiding future community-based conservation and classification studies of this important crop resource in the Caribbean. This is crucial to support the commercialization of breadfruit to improve its contribution to food and nutrition security.
Assuntos
Artocarpus , Fabaceae , Doença de Parkinson , Humanos , Artocarpus/genética , Etnobotânica , Conhecimento , BiodiversidadeRESUMO
Artocarpus incisa (breadfruit) seeds contain three different lectins (Frutalin, Frutapin (FTP) and Frutackin) with distinct carbohydrate specificities. The most abundant lectin is Frutalin, an α-D-galactose-specific carbohydrate-binding glycoprotein with antitumour properties and potential for tumour biomarker discovery as already reported. FTP is the second most abundant, but proved difficult to purify with very low yields and contamination with Frutalin frustrating its characterization. Here, we report for the first time high-level production and isolation of biologically active recombinant FTP in Escherichia coli BL21, optimizing conditions with the best set yielding >40 mg/l culture of soluble active FTP. The minimal concentration for agglutination of red blood cells was 62.5 µg/ml of FTP, a process effectively inhibited by mannose. Apo-FTP, FTP-mannose and FTP-glucose crystals were obtained, and they diffracted X-rays to a resolution of 1.58 (P212121), 1.70 (P3121) and 1.60 (P3121) Å respectively. The best solution showed four monomers per asymmetric unit. Molecular dynamics (MD) simulation suggested that FTP displays higher affinity for mannose than glucose. Cell studies revealed that FTP was non-cytotoxic to cultured mouse fibroblast 3T3 cells below 0.5 mg/ml and was also capable of stimulating cell migration at 50 µg/ml. In conclusion, our optimized expression system allowed high amounts of correctly folded soluble FTP to be isolated. This recombinant bioactive lectin will now be tested in future studies for therapeutic potential; for example in wound healing and tissue regeneration.
Assuntos
Artocarpus/genética , Expressão Gênica , Glucose/química , Manose/química , Lectinas de Plantas , Clonagem Molecular , Cristalografia por Raios X , Escherichia coli/química , Escherichia coli/genética , Escherichia coli/metabolismo , Lectinas de Plantas/biossíntese , Lectinas de Plantas/química , Lectinas de Plantas/genética , Domínios ProteicosRESUMO
Artocarpus heterophyllus Lam., commonly called jackfruit, is a medium-sized evergreen tree that bears high yields of the largest known edible fruit. Yet, it has been little explored commercially due to wide variation in fruit quality. The genetic diversity and genetic relatedness of 50 jackfruit accessions were studied using amplified fragment length polymorphism markers. Of 16 primer pairs evaluated, eight were selected for screening of genotypes based on the number and quality of polymorphic fragments produced. These primer combinations produced 5976 bands, 1267 (22%) of which were polymorphic. Among the jackfruit accessions, the similarity coefficient ranged from 0.137 to 0.978; the accessions also shared a large number of monomorphic fragments (78%). Cluster analysis and principal component analysis grouped all jackfruit genotypes into three major clusters. Cluster I included the genotypes grown in a jackfruit region of Karnataka, called Tamaka, with very dry conditions; cluster II contained the genotypes collected from locations having medium to heavy rainfall in Karnataka; cluster III grouped the genotypes in distant locations with different environmental conditions. Strong coincidence of these amplified fragment length polymorphism-based groupings with geographical localities as well as morphological characters was observed. We found moderate genetic diversity in these jackfruit accessions. This information should be useful for tree breeding programs, as part of our effort to popularize jackfruit as a commercial crop.
Assuntos
Artocarpus/classificação , Artocarpus/genética , Variação Genética , Análise do Polimorfismo de Comprimento de Fragmentos Amplificados , DNA de Plantas/genética , ÍndiaRESUMO
KM+, a mannose-binding lectin present in the seeds of Artocarpus integrifolia, has interesting biological properties and potential pharmaceutical use [A. Panunto-Castelo, M.A. Souza, M.C. Roque-Barreira, J.S. Silva, KM(+), a lectin from Artocarpus integrifolia, induces IL-12 p40 production by macrophages and switches from type 2 to type 1 cell-mediated immunity against Leishmania major antigens, resulting in BALB/c mice resistance to infection, Glycobiology 11 (2001) 1035-1042. ; L.L.P. daSilva, A. Panunto-Castelo, M.H.S. Goldman, M.C. Roque-Barreira, R.S. Oliveira, M.D. Baruffi, J.B. Molfetta-Machado, Composition for preventing or treating appearance of epithelia wounds such as skin and corneal wounds or for immunomodulating, comprises lectin, Patent number WO20041008.]. Here, we have isolated clones encoding the full-length KM+ primary sequence from a cDNA library, through matrix PCR-based screening methodology. Analysis of KM+ nucleotide and deduced amino acid sequences provided strong evidence that it neither enters the secretory pathway nor undergoes post-translational modifications, which is in sharp contrast with jacalin, the more abundant lectin from A. integrifolia seeds. Current investigations into the KM+ properties are often impaired by the difficulty in obtaining sufficient quantities of jacalin-free KM+ through direct seed extraction. To obtain active recombinant protein (rKM+) in larger amounts, we tested three different expression systems. Expression vectors were constructed to produce: (a) rKM+ in E. coli in its native form, (b) rKM+ with GST as an N-terminal tag and (c) native rKM+ in Saccharomyces cerevisiae. The presence of the GST-tag significantly improved the overall rKM+ yield; however, most of the obtained rGST-KM+ was insoluble. Production of rKM+ in the yeast host yielded the highest quantities of soluble lectin that retained the typical high-mannose oligosaccharide-binding properties of the natural protein. The possible biotechnological applications of recombinant KM+ are discussed.