Purification of a PHA-Like chitin-binding protein from Acacia farnesiana seeds: A time-dependent oligomerization protein

A lectin-like protein from the seeds of Acacia farnesiana was isolated from the albumin fraction, characterized, and sequenced by tandem mass spectrometry. The albumin fraction was extracted with 0.5 M NaCl, and the lectin-like protein of A. farnesiana (AFAL) was purified by ion-exchange chromatography (Mono-Q) followed by chromatofocusing. AFAL agglutinated rabbit erythrocytes and did not agglutinate human ABO erythrocytes either native or treated with proteolytic enzymes. In sodium dodecyl sulfate gel electrophoresis under reducing and nonreducing conditions, AFAL separated into two bands with a subunit molecular mass of 35 and 50 kDa. The homogeneity of purified protein was confirmed by chromatofocusing with a pI=4.0+/-0.5. Molecular exclusion chromatography confirmed time-dependent oligomerization in AFAL, in accordance with mass spectrometry analysis, which confers an alteration in AFAL affinity for chitin. The protein sequence was obtained by a liquid chromatography quadrupole time-of-flight experiment and showed that AFAL has 68% and 63% sequence similarity with lectins of Phaseolus vulgaris and Dolichos biflorus, respectively.

Purification, Characterization, and Preliminary X-Ray Diffraction Analysis of a Lactose-Specific Lectin from Cymbosema roseum Seeds

The unique carbohydrate-binding property of lectins makes them invaluable tools in biomedical research. Here, we report the purification, partial primary structure, carbohydrate affinity characterization, crystallization, and preliminary X-ray diffraction analysis of a lactose-specific lectin from Cymbosema roseum seeds (CRLII). Isolation and purification of CRLII was performed by a single step using a Sepharose-4B-lactose affinity chromatography column. The carbohydrate affinity characterization was carried using assays for hemagglutination activity and inhibition. CRLII showed hemagglutinating activity toward rabbit erythrocytes. O-glycoproteins from mucine mucopolysaccharides showed the most potent inhibition capacity at a minimum concentration of 1.2 A mu g mL(-1). Protein sequencing by mass spectrometry was obtained by the digestion of CRLII with trypsin, Glu-C, and AspN. CRLII partial protein sequence exhibits 46% similarity with the ConA-like alpha chain precursor. Suitable protein crystals were obtained with the hanging-drop vapor-diffusion method with 8% ethylene glycol, 0.1 M Tris-HCl pH 8.5, and 11% PEG 8,000. The monoclinic crystals belong to space group P2(1) with unit cell parameters a = 49.4, b = 89.6, and c = 100.8 A....

The action of the testa upon the germination of seeds of Cucumis anguria L.

In negatively photoblastic seeds of Cucumis anguria L. the scarification of micropyle overcame the inhibitory effect of white light , although far-red radiation inhibited the germination in both intact and scarified seeds. The moistened testa powder transmitted more far-red than red radiation. Thus the testa could act as a filter of radiation and maintain a high far-red/red ratio at the photosensitive site of the embryo, which must be localized near the micropyle. © 1988 Academia.

Electrical conductivity of soybean soaked seeds. I. Effect of genotype

During the period from October/92 to September/94 experiments were carried out at the Seed Laboratory, FCAV/UNESP, Jaboticabal, SP, Brazil, using soybean seeds of different genotypes in order to evaluate the effect of genotype on the electrical conductivity (bulk conductivity) of soaked seeds. Seed moisture content (105 ± 3°C, 24 h), standard germination (four 50-seed samples, paper towel, 30°C), and vigor-accelerated aging (42°C, 48 h) were first determined. Undamaged soybean seeds were soaked in deionized water (four 50-seed samples, 75 ml, 25°C, 24 h) and electrical conductivity (μmhos.cm+1.g+1) was measured. Significant differences in conductivity were observed among genotypes having the same pattern of germination and vigor. The results have showed that electrical conductivity can be significantly influenced by genotype.

Oxalate determination in urine using an immobilized enzyme on Sorghum vulgare seeds in a flow injection conductimetric system

A flow-injection (FI) method was developed for the determination of oxalate in urine. It was based on the use of oxalate oxidase (E.C. 1.2.3.4) immobilized on ground seeds of the BR-303 Sorghum vulgare variety. A reactor was filled with this activated material, and the samples (200 μL) containing oxalate were passed through it, carried by a deionized water flow. The carbon dioxide produced by the enzyme reaction permeated through a microporous PTFE membrane, and was received in a water acceptor stream, promoting conductivity changes proportional to the oxalate concentration in the sample. The results obtained showed a useful linear range from 0.05 to 0.50 mmol dm-3. The proposed method, when compared with the Sigma enzymatic procedure, showed good correlation (Y = 0.006(±0.016) + 0.98(±0.019)X; r = 0.9995, Y = conductivity in μS, and X = concentration in mmol dm-3), selectivity, and sensitivity. The new immobilization approach promotes greater stability, allowing oxalate determination for 6 months. About 13 determinations can be performed per hour. The precision of the proposed method is about ± 3.2 % (r.s.d).

Germinative pretreatments to dormancy break in Guazuma ulmifolia Lam. seeds

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Evaluation of naturally and artificially aged seeds of Phaseolus vulgaris L.

Seed ageing is natural phenomenon that occurs in all seeds, including those stored in dry and low temperature rooms. Phaseolus vulgaris L. cv. 'Carioca' seeds harvested in five different years were stored in at 15°C. Seed samples were germinated and evaluated according to The Brazilian Rules for Testing Seeds. The newest seed sample was submitted to artificial ageing. Small (∼1g) samples of all materials (naturally and artificially aged) were ground and proteins extracted. Equal quantities of protein were loaded onto the gels and electrophoresis carried out. Although seeds submitted to 24, 48, 72 and 96 hours of artificial ageing did not show any statistical difference (5%) in relation to unaged seeds in physiological parameters related to emergence, some statistically different parameters related to dry matter and length of shoots and roots occurred. A 24 h of artificial ageing at 41°C improved these factors. Protein patterns were changed after 72 h of artificial ageing and naturally aged seeds showed alterations after two years storage at 15°C. Aged seeds, naturally and artificially, had decreasing germination, vigour, and changes in the characteristic banding pattern of proteins. Physiological parameters and electrophoresis examination showed that for naturally aged seeds physiological parameters were more sensitive while artificially aged seeds, (41°C/100% RH), had electrophoretic profiles that were more efficient for seed lot discrimination.

Content of essential oil of fennel seeds in two different stages of development

This experiment was carried out at Plant Production Sector, Agronomical Science College-Botucatu, S.P., Brazil, in March, 2000. The aim of this assay was to determine the yield of essential oil of fennel (Foeniculum vulgare Miller) in different stages of development. Essential oils were prepared by hydrodistillation from the seeds using of Clevenger apparatus. The water utilized for the extraction of essential oil was sufficient to cover 100 g of seeds and the mixture was distilled for three hours. The volume of essential oil in the graduated side -arm of Clevenger apparatus was observed. There were no significative difference statistic was observed (Tukey 5%) in percentage (v/m) of oil content, based on dry weight of green seeds compared with dry weight of mature seeds, when they were harvested in two different stages of development. There was significative difference statistic between data obtained of humidity content of green seeds when these were compared with mature seeds. These results shows that others specifics studies about adaptation of fennel in tropical conditions are necessary, because the obtained data were different of data described on literature.

Phytochrome and light influence on germination and vigor of ocimum gratissimum L. (Lamiaceae) seeds under high irradiance conditions

Ocimum gratissimum seed germination (% germination and vigor) has been found as phytochrome dependent, having a typical High Irradiant Response (HIR). Seven treatments were tested: daylight (DL), red (R), far-red (FR), blue (B), green (G), dark (DK), and reversible (RVB). No statistical difference among the DL, R, FR, B, and G were found. DK and RVB were statistically equal and presented the lowest results. The germination also occurred in the DK treatment but in non-useful rates, and it was nonreversible in the RVB treatment. It allows these seeds to be classified as positively photoblastic. The minimum energy need to initiate the germination was evaluated by a fluency-response curve. It plotted four different exposition times to R light (1 second, 60 seconds, 1 hour, and 13 hours) against percent germination. Useful germination occurred only after 1 hour, confirming the high energy needed to incite the process. The germination rate increased with the raise of the photoequilibrium (j). The high positive correlation index found confirms the phytochrome influence in this process. Facing all the results presented here, it is suggested to sow these seeds under direct and highly intense sunlight. It is preferable to avoid places exposed to variations in the shading, because inhibition induced by dense shade effects (low R/FR ration and consequently low j established) were demonstrated irreversible, and it can lead to undesirable loss of the germination power.

Presence of Xylella fastidiosa in sweet orange fruit and seeds and its transmission to seedlings

Xylella fastidiosa, a xylem-limited bacterium, causes several economically important diseases in North, Central, and South America. These diseases are transmitted by sharpshooter insects, contaminated budwood, and natural root-grafts. X. fastidiosa extensively colonizes the xylem vessels of susceptible plants. Citrus fruit have a well-developed vascular system, which is continuous with the vascular system of the plant. Citrus seeds develop very prominent vascular bundles, which are attached through ovular and seed bundles to the xylem system of the fruit. Sweet orange (Citrus sinensis) fruit of cvs. Pera, Natal, and Valencia with characteristic symptoms of citrus variegated chlorosis disease were collected for analysis. X. fastidiosa was detected by polymerase chain reaction (PCR) in all main fruit vascular bundles, as well as in the seed and in dissected seed parts. No visual abnormalities were observed in seeds infected with the bacterium. However, the embryos of the infected seeds weighed 25% less than those of healthy seeds, and their germination rate was lower than uninfected seeds. There were about 2,500 cells of X. fastidiosa per infected seed of sweet orange, as quantified using real-time PCR techniques. The identification of X. fastidiosa in the infected seeds was confirmed by cloning and sequencing the specific amplification product, obtained by standard PCR with specific primers. X. fastidiosa was also detected in and recovered from seedlings by isolation in vitro. Our results show that X. fastidiosa can infect and colonize fruit tissues including the seed. We also have shown that X. fastidiosa can be transmitted from seeds to seedlings of sweet orange. To our knowledge, this is the first report of the presence of X. fastidiosa in seeds and its transmission to seedlings.

Productivity of beet and roquette cultivation as a function of time of establishing intercropping

With the aim of evaluating the effect of intercropping on the productivity of beet and roquette crops, a study was conducted in the Section of Vegetable and Aromatic-Medicinal Plants, FCAV-UNESP, Jaboticabal/SP. The treatments consisted of co-cultivations of roquette (Cultivada) and beets (Early Wonder) established at 0, 7, 14 and 21 days after the transplanting of beets, and as monocultures of beets and roquette. The experiments were delineated in a randomized block design. The productivity of beets in monoculture was not superior to that obtained with intercropping with roquette. The production of dry mass of roquette with intercropping was less than that with monoculture and the reduction was more marked with more delayed establishment of co-cultivation. Intercropping established at 0 and 7 days after the transplanting of beets was the most efficient, with indices of efficient use of land of 1.26 and 1.27, respectively.

Growth and yield analysis in different methods of beet (Cv. 'tall top early wonder') plantation

This objective of this study was to evaluate the effects of different methods of red beet seedling production and direct sowing on the development of the plant in field conditions. 'Tall Top Early Wonder' was used as the cultivar. The experimental design was a randomized blocks with 4 replications of each treatment: T1 - seedling produced in trays of 288 cells; T2 - 200 cells; T3 - 128 cells; T4 - 128 cells; T5 - direct sowing and T6 - bare-root seedling. The seedlings produced in trays and bare root seedlings were transplanted in the field 28 DAS with spacing of 0.20 x 0.10m. The mean height of plants, leaf area, leaf dry matter, petiole dry matter, shoot dry matter, root dry matter, relationship between shoot dry matter and root dry matter, absolute growth rate, relative growth rate, net assimilation rate, leaf area ratio and specific leaf area were determined. Storage root fresh matter and mean storage root diameter were determined starting from 77 DAS. Initial growth of the plants were superior for the direct sowing, resulting in smaller RGR and NAR than the other treatments. Independent of the production method, an increase of the cycle of the crop was verified. T6 had larger delay in the initial development. There was no difference for productivity. Method T1, had less expenses with respect to substrate and space in the vegetation home, without reduction in production.

Automatic extraction of road seeds from high-resolution aerial images

This article presents an automatic methodology for extraction of road seeds from high-resolution aerial images. The method is based on a set of four road objects and another set of connection rules among road objects. Each road object is a local representation of an approximately straight road fragment and its construction is based on a combination of polygons describing all relevant image edges, according to some rules embodying road knowledge. Each one of the road seeds is composed by a sequence of connected road objects, in which each sequence of this type can be geometrically structured as a chain of contiguous quadrilaterals. Experiments carried out with high-resolution aerial images showed that the proposed methodology is very promising in extracting road seeds. This article presents the fundamentals of the method and the experimental results, as well.

Storage in cerrado soil and germination of Psychotria vellosiana (Rubiaceae) seeds

The regeneration of plant communities from seed depends, to a large extent, on the capacity of the seed remaining viable in the soil. The viability and germination of artificially buried Psychotria vellosiana seeds in cerrado soil were studied, with the purpose of discovering some physio-ecological aspects of dispersed seeds and evaluating their potential to constitute a soil seed bank. Seed samples were placed in nylon envelopes and buried in the soil of a Cerrado reserve at two different depths and sites. Buried seeds were retrieved periodically and tested for germination along with dry-stored seeds. In general, there was a reduction in seed germination with storage time, both in soil and dry stored conditions, and in some assays exhumed seeds germinated faster than dry stored ones. In general the soil storage favoured seed viability of ungerminated seeds as compared to dry stored ones, with the seeds remaining partially viable after 10 months of storage. The lack of germination of viable seeds suggests that seeds showed true dormancy and/or required an extended time to germinate. It was observed that some seeds had germinated while buried and such in situ germination tended to increase with rainfall. The water availability in the soil might be a limiting factor for successful germination of P. vellosiana in the field, and the seeds may constitute a persistent soil seed bank in the cerrado as dispersed seeds remain viable in the soil until the following period of seed dispersal.