Public-domain software for root image analysis

In the search for high efficiency in root studies, computational systems have been developed to analyze digital images. ImageJ and Safira are public-domain systems that may be used for image analysis of washed roots. However, differences in root properties measured using ImageJ and Safira are supposed. This study compared values of root length and surface area obtained with public-domain systems with values obtained by a reference method. Root samples were collected in a banana plantation in an area of a shallower Typic Carbonatic Haplic Cambisol (CXk), and an area of a deeper Typic Haplic Ta Eutrophic Cambisol (CXve), at six depths in five replications. Root images were digitized and the systems ImageJ and Safira used to determine root length and surface area. The line-intersect method modified by Tennant was used as reference; values of root length and surface area measured with the different systems were analyzed by Pearson's correlation coefficient and compared by the confidence interval and t-test. Both systems ImageJ and Safira had positive correlation coefficients with the reference method for root length and surface area data in CXk and CXve. The correlation coefficient ranged from 0.54 to 0.80, with lowest value observed for ImageJ in the measurement of surface area of roots sampled in CXve. The IC (95 %) revealed that root length measurements with Safira did not differ from that with the reference method in CXk (-77.3 to 244.0 mm). Regarding surface area measurements, Safira did not differ from the reference method for samples collected in CXk (-530.6 to 565.8 mm²) as well as in CXve (-4231 to 612.1 mm²). However, measurements with ImageJ were different from those obtained by the reference method, underestimating length and surface area in samples collected in CXk and CXve. Both ImageJ and Safira allow an identification of increases or decreases in root length and surface area. However, Safira results for root length and surface area are closer to the results obtained with the reference method.

HEAVY METALS AND MICRONUTRIENTS IN THE SOIL AND GRAPEVINE UNDER DIFFERENT IRRIGATION STRATEGIES

Soils under natural conditions have heavy metals in variable concentrations and there may be an increase in these elements as a result of the agricultural practices adopted. Transport of heavy metals in soil mainly occurs in forms dissolved in the soil solution or associated with solid particles, water being their main means of transport. In this context, the aim of this study was to evaluate the heavy metal and micronutrient content in the soil and in the grapevine plant and fruit under different irrigation strategies. The experiment was carried out in Petrolina, PE, Brazil. The treatments consisted of three irrigation strategies: full irrigation (FI), regulated deficit irrigation (RDI), and deficit irrigation (DI). During the period of grape maturation, soil samples were collected at the depths of 0-10, 10-20, 20-40, 40-60, and 60-80 cm. In addition, leaves were collected at the time of ripening of the bunches, and berries were collected at harvest. Thus, the heavy metal and micronutrient contents were determined in the soil, leaves, and berries. The heavy metal and micronutrient contents in the soil showed a stochastic pattern in relation to the different irrigation strategies. The different irrigation strategies did not affect the heavy metal and micronutrient contents in the vine leaves, and they were below the contents considered toxic to the plant. In contrast, the greater availability of water in the FI treatment favored a greater Cu content in the grape, which may be a risk to vines, causing instability and turbidity. Thus, adoption of deficit irrigation is recommended so as to avoid compromising the stability of tropical wines of the Brazilian Northeast.

Organic and Nitrogen Fertilization of Soil under ‘Syrah’ Grapevine: Effects on Soil Chemical Properties and Nitrate Concentration

ABSTRACT Viticulture is an activity of great social and economic importance in the lower-middle region of the São Francisco River valley in northeastern Brazil. In this region, the fertility of soils under vineyards is generally poor. To assess the effects of organic and nitrogen fertilization on chemical properties and nitrate concentrations in an Argissolo Vermelho-Amarelo (Typic Plinthustalf), a field experiment was carried out in Petrolina, Pernambuco, on Syrah grapevines. Treatments consisted of two rates of organic fertilizer (0 and 30 m3 ha-1) and five N rates (0, 10, 20, 40, and 80 kg ha-1), in a randomized block design arranged in split plots, with five replications. The organic fertilizer levels represented the main plots and the N levels, the subplots. The source of N was urea and the source of organic fertilizer was goat manure. Irrigation was applied through a drip system and N by fertigation. At the end of the third growing season, soil chemical properties were determined and nitrate concentration in the soil solution (extracted by porous cups) was determined. Organic fertilization increased organic matter, pH, EC, P, K, Ca, Mg, Mn, sum of bases, base saturation, and CEC, but decreased exchangeable Cu concentration in the soil by complexation of Cu in the organic matter. Organic fertilization raised the nitrate concentration in the 0.20-0.40 m soil layer, making it leachable. Nitrate concentration in the soil increased as N rates increased, up to more than 300 mg kg-1 in soil and nearly 800 mg L-1 in the soil solution, becoming prone to leaching losses.

Comparison of citrus rooting evaluation methods using root images in soil profiles and root weight

A digitized image method was compared with a standard washing technique for measuring citrus roots in the field. Video pictures of roots were taken in a soil profile. The profile area analyzed was defined by iron rings, which were also used to remove the roots to determine their dry weight. The roots presented in the pictures were quantified using SIARCS software developed by Embrapa. The root length and area determined by digital images provided a good estimate of root quantity present in the profile.

Sample size for measurement of root traits on common bean by image analysis

Evaluation of root traits may be facilitated if they are assessed on samples of the root system. The objective of this work was to determine the sample size of the root system in order to estimate root traits of common bean (Phaseolus vulgaris L.) cultivars by digital image analysis. One plant was grown per pot and harvested at pod setting, with 64 and 16 pots corresponding to two and four cultivars in the first and second experiments, respectively. Root samples were scanned up to the completeness of the root system and the root area and length were estimated. Scanning a root sample demanded 21 minutes, and scanning the entire root system demanded 4 hours and 53 minutes. In the first experiment, root area and length estimated with two samples showed, respectively, a correlation of 0.977 and 0.860, with these traits measured in the entire root. In the second experiment, the correlation was 0.889 and 0.915. The increase in the correlation with more than two samples was negligible. The two samples corresponded to 13.4% and 16.9% of total root mass (excluding taproot and nodules) in the first and second experiments. Taproot stands for a high proportion of root mass and must be deducted on root trait estimations. Samples with nearly 15% of total root mass produce reliable root trait estimates.

Lipoxygenase activities during development of root and nodule of soybean

The objective of this work was to evaluate root and nodule soybean lipoxygenases in Doko cultivar and in a near isogenic line lacking seed lipoxygenases, inoculated and uninoculated with Bradyrhizobium elkanii. The lipoxygenase activities from roots collected at 3, 5, 9, 13, 18 and 28 days post-inoculation and from nodules collected at 13, 18 and 28 days post-inoculation were measured. The pH-activity profiles from root and nodules suggested that the lipoxygenases pool expressed in these organs from Doko cultivar and triple-null near isogenic lines are similar. The root lipoxygenase activity of Doko and triple-null lines, inoculated and uninoculated, reduced over time. The highest lipoxygenase activity observed at the beginning of root formation suggests the involvement of this enzyme in growth and development of this organ. However, for nodules an expressive increase of lipoxygenase activity was noticed 28 days post-inoculation. Root and nodule showed, at least, two mobility groups for lipoxygenases in immunoblottings, with approximately 94 and 97 kDa.

Reaction of arracacha genotypes to the root soft rot caused by Pectobacterium chrysanthemi

The purpose of this paper was to screen thirty-two arracacha genotypes for their reaction to root soft rot. Twenty roots of each genotype were inoculated with two Pectobacterium chrysanthemi isolates in a randomized experiment (10 roots/isolate). After inoculation, roots were individually wrapped with PVC film and kept at 26ºC in closed plastic bags. Soft rot lesions were recorded after 36 hours and genotypes were grouped in four classes of susceptibility by cluster analysis: 10 were less susceptible, 16 intermediate, 3 susceptible and 3 very susceptible. All the tested arracacha genotypes showed only variation in the degree of susceptibility.

Method for evaluation of root hairs of common bean genotypes

The objective of this work was to test a simple method for root hair evaluation of 21 common bean (Phaseolus vulgaris) genotypes, most of them used in breeding programs in Brazil. Hairs of basal and primary roots of 5-day old seedlings, produced on germination paper with no phosphorus addition, were visually evaluated by a rating scale after staining with 0.05% trypan blue. The method reveals variability among the genotypes, and the standard error of the mean is relatively low.

Root traits of common bean genotypes used in breeding programs for disease resistance

The objective of this work was to assess root traits of 19 common bean genotypes, used in breeding programs for disease resistance. Genotypes DOR 364 and G 19833 were used as deep and shallow basal root checks, respectively. The number of whorls and basal roots were assessed on five-day old seedlings grown in germination paper. Growth pouch studies were conducted to evaluate basal root gravitropism and lateral root length from primary roots, in seven-day old seedlings. The following root gravitropic traits were estimated: basal growth angle, shallow basal root length (localized in the top 2 cm), and relative shallow basal root growth. Number of whorls varied from 1.47 to 3.07, and number of basal roots ranged from 5.67 (genotype TO) to 12.07 (cultivar Jalo MG-65). Cultivars BRS MG Talismã, Carioca, BRS Pioneiro, and Diamante Negro exhibited shallow basal roots, while genotypes Vi-10-2-1, TU, AB 136, and México 54 showed deep basal roots. Cultivar Jalo MG-65 showed more lateral roots from the primary root than the other genotypes. Genotypes used on common bean breeding programs for disease resistance have great variability on basal and primary root traits.

Production of polyclonal antisera using recombinant coat proteins of Grapevine leafroll-associated virus 2 and Grapevine virus B

The objective of this work was to produce and characterize specific antisera against Brazilian isolates of Grapevine leafroll-associated virus 2 (GLRaV-2) and Grapevine virus B (GVB), developed from expressed coat proteins (CPs) in Escherichia coli, and to test their possible use for the detection of these two viruses in diseased grapevines. The coat protein (CP) genes were RT-PCR-amplified, cloned and sequenced. The CP genes were subsequently subcloned, and the recombinant plasmids were used to transform E. coli cells and express the coat proteins. The recombinant coat proteins were purified, and their identities were confirmed by SDS-PAGE and Western blot and used for rabbit immunizations. Antisera raised against these proteins were able to recognize the corresponding recombinant proteins in Western blots and to detect GLRaV-2 and GVB in infected grapevine tissues, by indirect ELISA, discriminating healthy and infected grapevines with absorbances (A405) of 0.08/1.15 and 0.12/1.30, respectively. Expressing CP genes can yield high amount of viral protein with high antigenicity, and GLRaV-2 and GVB antisera obtained in this study can allow reliable virus disease diagnosis.

Closed soilless growing system for producing strawberry bare root transplants and runner tips

The objective of this work was to test a closed soilless growing system for producing bare root transplants and runner tips of two strawberry clones, using two categories of substrates. The system used corrugated roofing panels of fiber-cement, over which a substrate layer was used as a growing bed. The nutrient solution was pumped from a reservoir toward the upper end of the roofing panels and drained back to a reservoir. Plant growth and development were determined for two advanced strawberry clones, grown in sand or in Plantmax organic substrate. Growth of the stock plants and the number and dry mass of bare root transplants were similar in the substrates, but bare roots differed in their crown diameters by substrate. For number of runner tips, no significant differences were found in total, small, and medium categories in the substrates. A mean production of about 590 runner tips per square meter and 145 bare root transplants per square meter was obtained. For both clones, a large number of bare root transplants and runner tips of adequate size were produced in the closed soilless growing system using sand or organic substrate.

Molecular characterization of grapevine from Santa Catarina, Brazil, using microsatellite markers

The objective of this work was to characterize the grape germplasm in Santa Catarina, Brazil, using microsatellite DNA markers (simple sequence repeats - SSR). The DNA samples were collected from leaves and shoots of accessions of public and private collections from the counties Urussanga, Nova Trento, Rodeio, São Joaquim, Campos Novos, Videira, and Água Doce. Ten SSR loci (VVS2, VVMD5, VVMD7, VVMD27, VrZAG62, VrZAG79, VVMD25, VVMD28, VVMD31, and VVMD32) were analysed by capillary electrophoresis. Molecular profiling was conducted for 190 grapevines (European, American, and hybrids), and 67 genotypes were obtained. The data were compared with each other and with those from the literature and from online databases, in order to identify varieties and discover cases of synonymy and homonymy. Forty molecular profiles corresponded to known varieties, while 27 genotypes were described for the first time. The existence of typical germplasm composed mainly of American and hybrid varieties is an important finding for local viticulture. Applications of the results rely on quality control and certification at the nursery level. Increasing precision in the characterization of grapevine genotypes may help breeding programs.

Leaf mineral composition of grafted eggplant grown in soil infested with Verticillium and root-knot nematodes

The objective of this work was to determine differences in leaf mineral composition between ungrafted and grafted onto (Solanum torvum) eggplant (Solanum melongena), cultivars 'Faselis' and 'Pala', grown in a soil infested with Verticillium dahliae and Meloidogyne incognita, or in a noninfested soil. Grafting increased leaf P and Mn concentrations, and decreased N concentrations, in both soils. Grafting also enhanced leaf Ca concentration of 'Pala', but it did not affect that of 'Faselis' depending on the cropping year. Leaf Mg concentration of grafted plants in infested soil was lower than that of ungrafted ones in noninfested soil. Results showed that, under the same fertilization program, the grafted 'Faselis' plants used the nutrients more efficiently than the 'Pala' ones. Use of S. torvum as a rootstock for 'Faselis' resulted in an effective protection against multiple pathogen infestation. Fertilization may be necessary when grafted 'Faselis' plants are grown in a soil infested with the pathogens, since grafting and infestation generally decrease leaf N, Mg, Ca and Fe concentrations, either by reducing the nutrient concentrations directly or by increasing leaf Mn concentration.

Obtaining interspecific hybrids, and molecular analysis by microsatellite markers in grapevine

The objective of this work was to assess the potential of interspecific hybridization of Vitis labruscana and Muscadinia rotundifolia by using artificial cross-pollinations. Microsatellite markers were used to confirm interspecific hybridizations and the identity of the parental genotypes. In crosses in which M. rotundifolia was used as the female parent, no true hybrids were obtained. In the reciprocal crosses, 114 seedlings were identified as true V. labruscana x M. rotundifolia hybrids. Self pollination occurred in direct and in reciprocal crosses. The crossings between 'Bordo' x 'Carlos', 'Magnolia', 'Regale' and' Roanoke', and between' Isabel' x 'Bountiful', 'Carlos', 'Magnolia', 'Regale' and 'Roanoke' were confirmed. The 15 markers evaluated showed that two M. rotundifolia parental genotypes had the same fingerprint profile, indicating a like lyplanting error. The success of hybridization depends mainly on the species and on the cultivar used as the female parent. Microsatellite markers are efficient to confirm the paternity of interspecific F1 hybrids and to determine the correct identity of M. rotundifolia cultivars.

Shoot growth of Merlot and Cabernet Sauvignon grapevine varieties

The objective of this work was to evaluate shoot growth of the grapevine varieties Merlot and Cabernet Sauvignon, during 2006/2007, and Cabernet Sauvignon, during 2008/2009, in São Joaquim, SC, Brazil. The experiment was carried out in a commercial vineyard trained on a vertical trellis system. The shoots of the central part of the plants were selected, and the lengths from the base to the apex of 20 shoots per cultivar were evaluated. In 2006/2007, monitoring began at pruning, on 9/15/2006, and ended on 2/6/2007, totalizing 144 days of evaluation. During the 2008/2009 cycle, phenology and shoot growth for 'Cabernet Sauvignon' were assessed from grape development (1/13/2009) (pea-sized grapes) until shoot vegetative growth had ceased. Budburst occurred in the second half of September, and shoot-growth cessation occurred during ripening. Higher growth rates (about 4 cm per day) were observed in pre- and post-flowering, followed by reduction due to the competition for photosynthates for the formation of flowers and bunches. Temperature and photoperiod induce grapevine shoots to cease growth in the highland regions of Santa Catarina State, Brazil.