Felix Camarena

Background: γ-Conglutin is a hemagglutinating lectin found in the seeds of the Andean legume Lupinus mutabilis. Leishmania (Viannia) peruviana is the etiological agent of cutaneous leishmaniasis, which is endemic in Peru. Leishmaniasis is a neglected disease that lacks an effective and non-toxic treatment. The aim of this study was to evaluate the effect of γ-conglutin from L. mutabilis Patón Grande ecotype (CoPG) on human monocyte cultures before infection with L. (V.) peruviana and to determine the production of molecules that stimulate cellular immunity. Methods: CoPG was isolated by chromatography from a hexane extract of L. mutabilis seeds. Monocytes were obtained by density gradient centrifugation. The effect of CoPG on their viability was determined by the MTT assay. To assess superoxide and nitric oxide production, monocyte cultures were pre-treated with CoPG and then infected with L. (V.) peruviana. Transcriptional expression of TNF-α and IL-1α in CoPG-treated and parasite-infected monocytes was determined by RT-PCR. Result: CoPG retained its hemagglutinating activity throughout the isolation process. The purified protein had a molecular weight of 47 KDa. Besides, CoPG at a 20 µg mL-1 induced the production of superoxide on the treated monocytes. This CoPG concentration also promoted the overexpression of IL-1α and TNF-α on the infected monocytes. The present study showed that CoPG has a novel in vitro immunomodulatory effect. The discovery of bioactive proteins from L. mutabilis may uncover a wide range of new applications for this Andean plant.

Lupinus mutabilis Sweet (Fabaceae), “tarwi” or “chocho”, is an important grain legume in the Andean region. In Peru, studies on tarwi have mainly focused on morphological features; however, they have not been molecularly characterized. Currently, it is possible to explore the genetic parameters of plants with reliable and modern methods such as genotyping by sequencing (GBS). Here, for the first time, we used single nucleotide polymorphism (SNP) markers to infer the genetic diversity and population structure of 89 accessions of tarwi from nine Andean regions of Peru. A total of 5922 SNPs distributed along all chromosomes of tarwi were identified. STRUCTURE analysis revealed that this crop is grouped into two clusters. A dendrogram was generated using the UPGMA clustering algorithm and, like the principal coordinate analysis (PCoA), it showed two groups that correspond to the geographic origin of the tarwi samples. AMOVA showed a reduced variation between clusters (7.59%) and indicated that variability within populations is 92.41%. Population divergence (F_st) between clusters 1 and 2 revealed low genetic difference (0.019). We also detected a negative F_is for both populations, demonstrating that, like other Lupinus species, tarwi also depends on cross-pollination. SNP markers were powerful and effective for the genotyping process in this germplasm. We hope that this information is the beginning of the path towards a modern genetic improvement and conservation strategies of this important Andean legume.

Background: Lupinus mutabilis Sweet is a species of Andean legume with a high content of proteins, which present hemagglutinating lectins. Here, we demonstrated that the γ-conglutin lectin isolated from L. mutabilis seeds is responsible for the hemagglutinating activity by evaluating this activity at each step of the isolation process. Methods: Seeds of three ecotypes of L. mutabilis were used. Saline protein extraction size exclusion and ionic exchange chromatography were performed to isolate the lectin. Carbohydrate specificity, thermostability and resistance to chelating and reducing agents of the lectin were tested. SDS-page and mass spectrometry were performed to characterize the isolated hemagglutinating lectin. Rabbit erythrocyte hemagglutination test was performed at each step. Result: Patón Grande ecotype had higher hemagglutinating titers and therefore was selected for further purification steps. Hemagglutinating activity of the purified lectin, which was identified as a γ-conglutin, was cation-independent and optimal between 15-20°C. Besides, it resisted temperatures up to 70°C, its activity was lost in basic pH and remained active under reducing and chelating conditions.

Lupinus mutabilis Sweet (Fabaceae), “tarwi” or “chocho”, is an important grain legume in the Andean region. In Peru, studies on tarwi have been mainly focused on morphological features, however, the have not been molecularly characterized. Currently, it is possible to explore genetic parameters of plants with reliable and modern methods like genotyping-by-sequencing (GBS). We here for the first time used single nucleotide polymorphisms (SNPs) markers to infer the genetic diversity and population structure of 89 accessions of tarwi from nine Andean regions of Peru. A total of 5922 SNPs distributed along all chromosomes of tarwi were identified. STRUCTURE analysis revealed that this crop is grouped into two clusters. A dendrogram was generated using the UPGMA clustering algorithm and, similar to the principal coordinate analysis (PCoA), it showed two groups that correspond to the geographic origin of the tarwi samples. AMOVA showed a reduced variation between clusters (7.59 %) and indicated that variability within populations is 92.41 %. Population divergence (Fst) between clusters 1 and 2 revealed low genetic difference (0.019). We also detected a negative Fis for both populations, demonstrating that, similar to other Lupinus species, tarwi also depends on cross-pollination. SNPs markers were powerful and effective for the genotyping process in this germplasm. We hope that this information is the beginning of the path towards a modern genetic improvement and conservation strategies of this important Andean legume.

Tarwi (Lupinus mutabilis Sweet) is an Andean legume that has attracted international interest due to its high nutritional value. This has resulted in an increase in its conventional production, which leads to an ecological imbalance. In this context, the application of biotechnologies, based on the use of bacterial inoculants, is of utmost importance. This work aimed to evaluate the effects of a consortium of 2 strains of Bradyrhizobium spp. (BR) and 1 strain of Bacillus sp. (BA) on tarwi var. Andenes. The treatments tested were BR + BA, BR + Organic Matter, BR + Agrochemical (Azoxystrobin y Difenoconazole), and the control (without application). The crop was located in Marcara-Ancash (altitude 3254 masl), Peru. The experiment involved the inoculation of BR in the seeds and a re-inoculation 30 days later. BA was inoculated every 30 days in the neck of the plant and aerial part, 5 times during plant development. The inoculation with BR + BA significantly increased the aerial fresh weight (413.2%), plant height (13.5%), and diminished the anthracnose (38.4%) of plants 110 DAS (days after the sowing). Also, this treatment produced the best-harvested emergence percentage (97.9%), morpho-agronomic characteristics, and an increase in the yield (171%) compared to the control. In conclusion, the application of the Bacillus sp. strain and the Bradyrhizobium spp. consortia improved the productivity of tarwi var. Andenes. The interaction of these strains have the potential to be used in tarwi field planting programs.

Lupinus mutabilis protein-rich seeds must be debittered before consumption. The aim of this research was to assess free phenolic compounds, antioxidant capacity and FT-NIR spectra of flours from debittered seeds of 33 Andean ecotypes of L. mutabilis, and five varieties belonging to L. luteus, L. angustifolius and L. albus, as controls. The free phenolics were quantified by RP-HPLC, while the antioxidant capacity was evaluated spectrophotometrically through the Reducing Power, ABTS, FRAP and DPPH methods. The free phenolics of L. mutabilis were mostly (85.5–99.6%) flavonoids (genistein and genistein derivatives, apigenin, catechin and naringenin). Other compounds, detected in low quantities, were phenylethanoids (tyrosol and tyrosol derivative) and phenolic acids (cinnamic acid derivatives). The highest total free phenolic concentration was observed in H6 INIA BP (1393.32 mg/kg DM), followed by Chacas, Moteado beige, Huánuco and Lircay. The antioxidant capacity of the L. mutabilis ecotypes exceeded that of the controls and was correlated to flavonoids content. Additionally, a relationship between free phenolic compounds and spectral bands was established by FT-NIR, paving the way for a fast, reliable and non-destructive approach to lupin seeds characterisation. Even after debittering, lupin flours maintained high free phenolic concentrations and antioxidant capacity.

SOSTENIBILIDAD SOCIAL DE LOS SUBSISTEMAS PRODUCTIVOS DE TOMATE DE ÁRBOL (Solanum betaceum Cav) EN EL CANTÓN GUACHAPALA, PROVINCIA DE AZUAY - ECUADOR