Effect of arcelin protein on the biology of Zabrotes subfasciatus (Boheman 1833), in dry beans

Arcelin is a seed protein found in wild beans (Phaseolus vulgaris) which gives resistance to Mexican bean weevil, Zabrotes subfasciatus (Boheman 1833) (Coleoptera: Bruchidae). Studies were carried out with the objective of estimating the effect of four alleles of protein arcelin (Arc1, Arc2, Arc3 and Arc4) on the biology of Z. subfasciatus. The experiment was carried out in laboratory at Embrapa-Centro Nacional de Pesquisa de Arroz e Feijão, in Santo Antônio de Goiás, GO, Brazil, under non controlled conditions. The highest levels of antibiosis to Z. subfasciatus were observed in Arc1, with reduction in the number of eggs, number of emerged adults, adults longevity. In the line Arc2 only reduction in the number of emerged adults was observed. The lines Arc3 and Arc4 showed low efficiency on the reduction of progeny of Z. subfasciatus and effects in the longevity and egg-adult cycle were not detected. Insect sexual ratio was not altered by the presence of Arc1, Arc2, Arc3 and Arc4 in the seeds.

Variability of ricin content in mature seeds of castor bean

The objective of this work was to evaluate ricin concentration in castor bean seeds (Ricinus communis) of 20 accessions from the Banco de Germoplasma de Mamoneira of the Embrapa Algodão, Campina Grande, PB, Brazil, using the Enzyme Linked Immunosorbent Assay. Significant differences were observed among accessions. BRA 3271 had the highest ricin concentration in seeds (32.18 ng µg-1), and BRS Paraguaçu had the lowest (3.53 ng µg-1). There is the possibility of selecting genotypes with different ricin concentrations, which can be used according on the interest of the breeding programs.

Growth, nodulation, and yield of soybean and associated weeds as affected by weed management

Two field experiments were conducted at the experimental farm of the National Research Centre at Shalakan, Kalubia Governorate, Egypt, during 2006 and 2007 seasons, to study the effect of three pre-emergence herbicides, prometryn [at the rate of 0.75, 1.5 and 2.25 kg ha-1], oxadiargyl [at the rate of 240, 480 and 720 g ha-1] and butralin [at the rate of 1.20, 2.40 and 3.36 kg ha-1], two hand hoeing treatments and a nonweeded check, on weed infestation, nodulation, growth, yield and yield attributes of soybean plants. Two hand hoeing treatments resulted in the highest weed depression expressed as the lowest fresh and dry weights of broadleaved, grassy and total weeds. The reduction percentage in weed dry matter compared to the nonweeded treatment was 98.3, 92.64 and 96.9% in broadleaved, grassy and total weeds, respectively. Application of the three herbicides at higher or recommended doses significantly reduced fresh and dry weight of the weeds compared to the nonweeded treatment. The results indicated that all the three herbicides at rates higher than the recommended markedly decreased the number, fresh and dry weight of nodules as well as root, shoot and total dry weight plant-1, while application of two hand hoeing treatments significantly increased these traits. Two hand hoeing treatments and pre-emergence herbicides at the recommended rates markedly increased soybean yield and its attributes. Two hand hoeing treatments gave the highest values of number of pods per plant-1, weight of pods per plant-1 and number of seeds per plant-1 by 140.7, 150.0 and 59.8%, respectively, compared to the nonweeded treatment. On the other hand, oxadiargyl at the recommended rate (480 g ha-1) was the best treatment for promoting seed yield (g plant-1), seed yield (kg ha-1) and biological yield (g plant-1) compared to the nonweeded treatment by 87.3, 85.0 and 88.2%, respectively. Prometryn at the rate of 1.50 kg ha-1, followed by two hand hoeing treatments, produced the highest shoot and seed protein percentage as well as seed oil percentage, compared to the other weed control treatments.

Nutritional quality and bioactive compounds of partially defatted baru almond flour

This study aimed to investigate the nutritional quality and bioactive potential of partially defatted baru (Dipteryx alataVog.) almond flour (BAF). The flour’s proximate and mineral compositions, total phenolic, tocopherols and carotenoids contents, antioxidant capacity, trypsin inhibitor and amino acid analyses were performed. An experiment was conducted with 24 male Wistar rats in order to evaluate the flour’s protein quality. BAF has high protein, fiber and mineral contents (iron, zinc, magnesium and copper), and it is a source of calcium. BAF presented relevant amounts of total phenolics (625 mg/100g) and good antioxidant capacity (130 µmol/Trolox eq). Autoclaved BAF showed essential amino acids profile, digestibility and protein quality better than in natura BAF. Autoclaved BAF might be used for human consumption as a source of quality protein and bioactive compounds, in healthy diets and processed foods.

Grain yield and seed quality of soybean selected for high protein content

The objective of this work was to evaluate the effects of selection for high protein on seed physiological quality and grain yield of soybean. Four populations of BC1F4 and four of F4, each from a cross between a commercial variety and a line bearing high protein seeds, were used. The high protein content selection has a tendency to affect negatively the seed physiological quality. Estimates of correlation coefficients between protein content and grain yield were mostly negative but varied among populations. It is possible to obtain lines with high protein content, keeping the grain yield and the seed physiological quality of their respective recurrent progenitors.

Subunit composition of seed storage proteins in high-protein soybean genotypes

The objective of this work was to quantify the accumulation of the major seed storage protein subunits, β-conglycinin and glycinin, and how they influence yield and protein and oil contents in high-protein soybean genotypes. The relative accumulation of subunits was calculated by scanning SDS-PAGE gels using densitometry. The protein content of the tested genotypes was higher than control cultivar in the same maturity group. Several genotypes with improved protein content and with unchanged yield or oil content were developed as a result of new breeding initiatives. This research confirmed that high-protein cultivars accumulate higher amounts of glycinin and β-conglycinin. Genotypes KO5427, KO5428, and KO5429, which accumulated lower quantities of all subunits of glycinin and β-conglycinin, were the only exceptions. Attention should be given to genotypes KO5314 and KO5317, which accumulated significantly higher amounts of both subunits of glycinin, and to genotypes KO5425, KO5319, KO539 and KO536, which accumulated significantly higher amounts of β-conglycinin subunits. These findings suggest that some of the tested genotypes could be beneficial in different breeding programs aimed at the production of agronomically viable plants, yielding high-protein seed with specific composition of storage proteins for specific food applications.

Influence of wheat straw and rhizosphere on seed germination, early seedling growth and bio-chemical attributes of Trianthema portulacastrum

Decomposing wheat (Triticum aestivum) straw and rhizosphere-infested soil were evaluated for their suppressive activity against horse purslane (Trianthema portulacastrum), a noxious summer weed in Pakistan. Two separate pot studies were carried out. Wheat straw was incorporated at 4, 6 and 8 g kg-1 soil five days before the sowing of horse purslane. Pots without straw incorporation were maintained as control. In a second study, soil was taken from 15 and 30 cm depths from a previously cropped wheat field immediately after its harvest and was used as growing medium. Soil from an intentionally uncropped area of the same field was used as control. Suppressive activity was measured in terms of germination dynamics, seedling growth, and biochemical attributes such as chlorophyll contents, total soluble phenolics, soluble protein and antioxidant enzymes. Germination, seedling growth, chlorophyll contents and soluble protein of horse purslane were all negatively influenced. Higher phenolics and enhanced activities of antioxidant enzymes were noticed in response to wheat residues incorporation and its rhizosphere soil. Both studies established that the phytotoxic influence of wheat straw and wheat-infested rhizosphere soil on horse purslane can further be exploited for horse purslane management as a sustainable approach.

Brazilian soybean Glycine max (L.) Merr. cultivars adapted to low latitude regions: seed composition and content of bioactive proteins

Among the goals of the Brazilian soybean improvement programmes, the breeding strategies for cultivars adapted to low latitudes have been included to extend crop areas and to increase production. Seeds of nine Brazilian soybean cultivars adapted to low latitudes were investigated regarding to their composition, and amino acid and antinutritional/toxic protein contents. Protein (394.5 ± 13.1 to 445.3 ± 8.0 g kg-1 dry matter) and oil (200.6 ± 1.2 to 232.3 ± 4.7 g kg-1 dry matter) contents showed low correlation to each other (r = -0.06). The total carbohydrate (141.7 ± 6.1 to 211.1 ± 15.0 g kg-1 dry matter) and ash contents (48.2 ± 4.2 to 52.2 ± 0.5 g kg-1 dry matter) were similar to data available for other soybean cultivars. All soybean cultivars presented low levels of tryptophan and sulphur amino acids. The lectin (1,152 to 147,456 HU kg-1 flour), trypsin inhibitor (34.45 ± 2.28 to 77.62 ± 2.63 g trypsin inhibited kg-1 flour), toxin (6,210 ± 134 to 34,650 ± 110 LD50 kg-1 flour) and urease (0.74 ± 0.02 to 1.22 ± 0.10 g kg¹ flour) presented variations in their contents amongst the cultivars. Compared to other soybean cultivars, urease was higher, the acute toxicity lower and the lectin and trypsin inhibitor contents similar to data available. In general, soybean cultivars showed similar biochemical composition to those developed in different geographic regions. The relevance of these findings to the agronomic features and to choice of soybean cultivars to be used as food or feed is discussed.

Isolation and characterization of a reserve protein from the seeds of Opuntia ficus-indica (Cactaceae)

We describe here the isolation and characterization of a major albumin from the seeds of Opuntia ficus-indica (Cactaceae). This protein has a molecular mass of 6.5 kDa and was isolated by a combination of gel filtration chromatography and reverse-phase HPLC. The amino acid composition of this protein was determined and it was shown to have similarities with the amino acid composition of several proteins from the 2S albumin storage protein family. The N-terminal amino acid sequence of this protein is Asp-Pro-Tyr-Trp-Glu-Gln-Arg.

Purification and partial characterization of Phaseolus vulgaris seed aminopeptidase

The aminopeptidase activity of Phaseolus vulgaris seeds was measured using L-Leu-p-nitroanilide and the L-aminoacyl-ß-naphthylamides of Leu, Ala, Arg and Met. A single peak of aminopeptidase activity on Leu-ß-naphthylamide was eluted at 750 µS after gradient elution chromatography on DEAE-cellulose of the supernatant of a crude seed extract. The effluent containing enzyme activity was applied to a Superdex 200 column and only one peak of aminopeptidase activity was obtained. SDS-polyacrylamide gel electrophoresis (10%) presented only one protein band with molecular mass of 31 kDa under reducing and nonreducing conditions. The aminopeptidase has an optimum pH of 7.0 for activity on all substrates tested and the highest Vmax/KM ratio for L-Leu-ß-naphthylamide. The enzyme activity was increased 40% by 0.15 M NaCl, inhibited 94% by 2.0 mM Zn2+, inhibited 91% by sodium p-hydroxymercuribenzoate and inhibited 45% by 0.7 mM o-phenanthroline and 30 µM EDTA. Mercaptoethanol (3.3 mM), dithioerythritol (1.7 mM), Ala, Arg, Leu and Met (70 µM), p-nitroaniline (0.25 mM) and ß-naphthylamine (0.53 mM) had no effect on enzyme activity when assayed with 0.56 mM of substrate. Bestatin (20 µM) inhibited 18% the enzyme activity. The aminopeptidase activity in the seeds decayed 50% after two months when stored at 4oC and room temperature. The enzyme is leucyl aminopeptidase metal- and thiol group-dependent.

Lima bean (Phaseolus lunatus) seed coat phaseolin is detrimental to the cowpea weevil (Callosobruchus maculatus)

The presence of phaseolin (a vicilin-like 7S storage globulin) peptides in the seed coat of the legume Phaseolus lunatus L. (lima bean) was demonstrated by N-terminal amino acid sequencing. Utilizing an artificial seed system assay we showed that phaseolin, isolated from both cotyledon and testa tissues of P. lunatus, is detrimental to the nonhost bruchid Callosobruchus maculatus (F) (cowpea weevil) with ED50 of 1.7 and 3.5%, respectively. The level of phaseolin in the seed coat (16.7%) was found to be sufficient to deter larval development of this bruchid. The expression of a C. maculatus-detrimental protein in the testa of nonhost seeds suggests that the protein may have played a significant role in the evolutionary adaptation of bruchids to legume seeds.

Characterization of a methionine-rich protein from the seeds of Cereus jamacaru Mill. (Cactaceae)

We describe here the isolation and characterization of a major albumin from the seeds of Cereus jamacaru (Cactaceae), to which we gave the trivial name of cactin. This protein has a molecular mass of 11.3 kDa and is formed by a light chain (3.67 kDa) and a heavy chain (7.63 kDa). This protein was isolated using a combination of gel filtration chromatography and reverse-phase HPLC. The amino acid composition of cactin was determined and found to resemble that of the 2S seed reserve protein from the Brazil nut, a protein remarkable for its high methionine content. The usefulness of cactin as a molecular marker in the taxonomy of the Cactaceae is discussed.

A protein with amino acid sequence homology to bovine insulin is present in the legume Vigna unguiculata (cowpea)

Since the discovery of bovine insulin in plants, much effort has been devoted to the characterization of these proteins and elucidation of their functions. We report here the isolation of a protein with similar molecular mass and same amino acid sequence to bovine insulin from developing fruits of cowpea (Vigna unguiculata) genotype Epace 10. Insulin was measured by ELISA using an anti-human insulin antibody and was detected both in empty pods and seed coats but not in the embryo. The highest concentrations (about 0.5 ng/µg of protein) of the protein were detected in seed coats at 16 and 18 days after pollination, and the values were 1.6 to 4.0 times higher than those found for isolated pods tested on any day. N-terminal amino acid sequencing of insulin was performed on the protein purified by C4-HPLC. The significance of the presence of insulin in these plant tissues is not fully understood but we speculate that it may be involved in the transport of carbohydrate to the fruit.

Identification of Albizia lebbeck seed coat chitin-binding vicilins (7S globulins) with high toxicity to the larvae of the bruchid Callosobruchus maculatus

Seed coat is a specialized maternal tissue that interfaces the embryo and the external environment during embryogenesis, dormancy and germination. In addition, it is the first defensive barrier against penetration by pathogens and herbivores. Here we show that Albizia lebbeck seed coat dramatically compromises the oviposition, eclosion and development of the bruchid Callosobruchus maculatus. Dietary supplementation of bruchid larvae with A. lebbeck seed coat flour causes severe weight loss and reduces survival. By means of protein purification, mass spectrometry and bioinformatic analyses, we show that chitin-binding vicilins are the main source of A. lebbeck tegumental toxicity to C. maculatus. At concentrations as low as 0.1%, A. lebbeck vicilins reduce larval mass from 8.1 ± 1.7 (mass of control larvae) to 1.8 ± 0.5 mg, which corresponds to a decrease of 78%. Seed coat toxicity constitutes an efficient defense mechanism, hindering insect predation and preventing embryo damage. We hypothesize that A. lebbeck vicilins are good candidates for the genetic transformation of crop legumes to enhance resistance to bruchid predation.

Thermal properties of defatted meal, concentrate, and protein isolate of baru nuts (Dipteryx alata Vog.)

Baru (Dipteryx alata Vog.), a species of legume found in the Brazilian savannas, was investigated in this study for the composition of its flesh and seed. Thermal analyses, Thermogravimetry (TG), and Differential Scanning Calorimetry (DSC) were used to investigate the proteins in defatted meal, concentrate, and protein isolate. The protein concentrate was extracted at pH 10, followed by a precipitation at the isoelectric point to obtain the isolate that was spray dried. The thermogravimetric curves were obtained under a nitrogen atmosphere with a 100 mL/minutes flow. The initial, final and peak temperatures and mass loss were analyzed. Within the performed temperature ranges studied, the defatted meal and concentrate presented four steps of mass loss, while the isolate showed only two steps. The protein content of defatted meal from Baru nuts was higher than that of the isolate. On the other hand, there was a reduction in enthalpy, which suggests that the process applied to obtain the baru concentrate and isolate led to protein denaturation.