A low-mass impact sensor for high-speed firmness sensing of fruits.

A low-mass impact sensor for high-speed firmness sensing of fruits was built and tested. Results of tests with a rubber ball indicated that the impact measurement was not sensitive to the distance between the impactor and the impacting surface of the sample within the range of 8 to 23 mm, and was not sensitive to how the sample was held. Tests with kiwifruits and peaches show good correlation between firmness readings obtained with the impact sensor and those obtained with the penetrometer. The best correlation was between the slope of the impact curve (at mid-point) and the force-deformation firmness. Preliminary test showed that the sensor could sense fruit firmness at a speed of 5 fruits/s.

Effect of impacting radius of curvature on firmness sensing of fruits in a sorting system.

Fruits of two varieties of both apples and pear were tested to measure their response to small energy impact applied by a impact tester with two spherical tips of different radious of curvature ( RA = 2.48 cm and RB = 0.98 cm) and equal mass were used. In the four varieties studied, the size of bruise was smaller with a spherical tip RA than with tip RB . The non-destructive impact test would cause less damage with a spherical impactor with a radious bigger than 0.98 cm.

Mealiness assessment in fruits using MRI techniques.

Mealiness is a sensory attribute that cannot be defined by a single parameter but through a combination of variables (multidimensional structure). Previous studies propose the definition of mealiness as the lack of crispiness, of hardness and of juiciness. Current aims are focused on establishing non destructive tests for mealiness assessment. MultiSliceMultiEcho Magnetic resonance images (MRI, 64*64pixels) have been taken corresponding to a 3ms of Echo time. Small samples of Top Red apples stored 6 months at controlled atmosphere (expected to be non mealy) and 2°C (expected to be mealy) have been used for MRI imaging. Three out of four apples corresponding to the sample maintained at controlled atmosphere did not develop mealiness while three out of four fruits corresponding to the sample stored at 2°C became mealy after 6 month of storage. The minimum T2 values/image obtained for the mealy apples shows to be significantly lower when compared with non mealy apples pointing that a more dis-aggregated structure leads to a quicker loss of signal Also, there is a significant linear correlation (r=-0.76) between the number of pixels with a T2 value below 35ms within a fruit image and the deformation parameter registered during the Magness-Taylor firmness test. Finally, all the T2 images of the mealy apples show a regional variation of contrast which is not shown for non mealy apples. This variation of contrast is similar to the MRI images of water-cored apples indicating that in these cases there is a differential water movement that may precede the internal browning.

A quality assessment system for fresh fruits in hypermarkets and fruit trading centres.

When the fresh fruit reaches the final markets from the suppliers, its quality is not always as good as it should, either because it has been mishandled during transportation or because it lacks an adequate quality control at the producer level, before being shipped. This is why it is necessary for the final markets to establish their own quality assessment system if they want to ensure to their customers the quality they want to sell. In this work, a system to control fruit quality at the last level of the distribution channel has been designed. The system combines rapid control techniques with laboratory equipment and statistical sampling protocols, to obtain a dynamic, objective process, which can substitute advantageously the quality control inspections carried out visually by human experts at the reception platform of most hypermarkets. Portable measuring equipment have been chosen (firmness tester, temperature and humidity sensors...) as well as easy-to-use laboratory equipment (texturometer, colorimeter, refractometer..,) combining them to control the most important fruit quality parameters (firmness, colour, sugars, acids). A complete computer network has been designed to control all the processes and store the collected data in real time, and to perform the computations. The sampling methods have been also defined to guarantee the confidence of the results. Some of the advantages of a quality assessment system as the proposed one are: the minimisation of human subjectivity, the ability to use modern measuring techniques, and the possibility of using it also as a supplier's quality control system. It can be also a way to clarify the quality limits of fruits among members of the commercial channel, as well as the first step in the standardisation of quality control procedures.

Non-destructive optical characterisation of fruits with time-resolved reflectance spectroscopy.

Increasing attention is being paid to the possible development of non-invasive tests for the assessment of the quality of fruits We propose a novel non-destructive method for the measurement of the internal optical properties of fruits and vegetables by means of time resolved reflectance spectroscopy in the visible and NIR range. A fully automated instrumentation for time-resolved reflectance measurements was developed It is based on mode-locked laser sources and electronics for time-correlated single photon counting, and provides a time-resolution of 120-160 ps The system was used to probe the optical properties of several species and varieties of fruits and vegetables in the red and NIR range (650-1000 nm). In most fruits, the absorption line shape is dominated by the absorption peak of water, centred around 970 nm Generally, the absorption spectra also show the spectral features typical of chlorophyll, with maximum at 675 nm In particular, for what concerns apples, variations in peak intensity are observed depending on the variety, the degree of ripeness as well as the position on the apple. For all the species and varieties considered, the transport scattering coefficient decreases progressively upon increasing the wavelength.

Non-destructive optical characterization of fruits in the red and near infrared.

Increasing attention is being paid to the possible development of non-invasive tests for the assessment of the quality of Fruits. We propose a novel non-destructive method for the measurement of the internal optical properties of fruits and vegetables by means of lime-resolved reflectance spectroscopy in the visible and NIR range. A Fully automated instrumentation for time-resolved reflectance measurements was developed. It is based on mode-locked laser sources and electronics for time-correlated single photon counting, and provides a time-resolution of 120-160 ps. The system was used to probe the optical properties of several species and varieties of Fruits and vegetables in the red and NIR range (650-1000 nm). In most Fruits, the absorption line shape is dominated by the absorption peak of water, centred around 970 nm. Generally, the absorption spectra also show the spectral features typical of chlorophyll, with maximum at 675 nm. In particular, for what concerns apples, variations in peak intensity are observed depending on the variety, the degree of ripeness as well as the position on the apple. For all the species and varieties considered, the transport scattering coefficient decreases progressively upon increasing the wavelength.

New version of a laboratory impact device for firmness sensing of fruits.

Results of previous studies conducted by different researchers have shown that impact techniques can be used to evaluate firmness (Delwiche et al., 1989; Delwiche et al.;1996; Jaren et al., 1992; Ruiz Altisent et al., 1996). To impact the fruit with a small spherical impactor of known mass and radius of curvature and measure the acceleration of the impactor is a technique described by Chen et al. (1985) and used by several researchers for sensing fruit firmness (Jaren et al., 1992; Correa et al.; 1992). The advantages of this method vs. a force sensor that measures the force as a function of time is that the measured impact-acceleration response is independent of the fruit mass and is less sensitive to the variation in the radius of curvature of the fruit (Chen et al., 1996). Ruiz Altisent et al. (1993) developed and used a 50 g impactor with a 19 mm diameter spherical tip, dropping from different height for fruits (apples, pears, avocados, melons, peaches ...). Another impact device for firmness sensing of fruits was developed by Chen and Ruiz Altisent (1996). They designed and fabricated an experimental low-mass impact sensor for high-speed sensing of fruit firmness. The impactor consisted of a semi-spherical impacting tip attached to the end (near the centre of percussion) of a pivoting arm. Impact is done by swinging the impactor to collide with the fruit. It has been implemented for on-line use. In both devices a small accelerometer is mounted behind the impacting tip. Lateral impactor and vertical impactor have been used in laboratory and the results from non-destructive impact tests have contributed to standardise methods to measure fruit firmness: Barreiro (1992) compared impact parameters and results of Magness-Taylor penetration tests for apples, pears, apricots [and peaches; Agulheiro (1994) studied the behaviour of the impact parameters during seven weeks of cold storage of two melon varieties; Ortiz (1998) used low energy impact and NIR procedures to segregate non crispy, non firm and soft peaches. Steinmetz (1996) compared various non-destructive firmness sensors, based on sound, impact and micro-deformation.

Lateral impact sensor for measuring firmness of fruits in an experimental packing line.

Different parameters are used to quantify the maturity of fruits at or near harvest (shape, color, flesh texture and internal composition). Flesh firmness is a critical handling parameter for fruits such as peach, pear and apple. Results of previous studies conducted by different researchers have shown that impact techniques can be used to evaluate firmness of fruits. A prototype impact system for firmness sorting of fruits was developed by Chen and Ruiz-Altisent (Chen et al, 1996). This sensor was mounted and tested successfully on a 3 m section of a commercial conveyor belt (Chen et al, 1998). This is a further development of the on-line impact system for firmness sorting of fruits. The design of the sensor has been improved and it has been mounted on a experimental fruit packing line (Ortiz-Cañavate et al 1999).

Expression of xyloglucan endotransglucosylase/hydrolase (XTH) genes and XET activity in ethylene treated apple and tomato fruits.

Xyloglucan endotransglucosylase/hydrolase (XTHs: EC 2.4.1.207 and/or EC 3.2.1.151), a xyloglucan modifying enzyme, has been proposed to have a role during tomato and apple fruit ripening by loosening the cell wall. Since the ripening of climacteric fruits is controlled by endogenous ethylene biosynthesis, we wanted to study whether XET activity was ethylene-regulated, and if so, which specific genes encoding ripening-regulated XTH genes were indeed ethylene-regulated. XET specific activity in tomato and apple fruits was significantly increased by the ethylene treatment, as compared with the control fruits, suggesting an increase in the XTH gene expression induced by ethylene. The 25 SlXTH protein sequences of tomato and the 11 sequences MdXTH of apple were phylogenetically analyzed and grouped into three major clades. The SlXTHs genes with highest expression during ripening were SlXTH5 and SlXTH8 from Group III-B, and in apple MdXTH2, from Group II, and MdXTH10, and MdXTH11 from Group III-B. Ethylene was involved in the regulation of the expression of different SlXTH and MdXTH genes during ripening. In tomato fruit fifteen different SlXTH genes showed an increase in expression after ethylene treatment, and the SlXTHs that were ripening associated were also ethylene dependent, and belong to Group III-B (SlXTH5 and SlXTH8). In apple fruit, three MdXTH showed an increase in expression after the ethylene treatment and the only MdXTH that was ripening associated and ethylene dependent was MdXTH10 from Group III-B. The results indicate that XTH may play an important role in fruit ripening and a possible relationship between XTHs from Group III-B and fruit ripening, and ethylene regulation is suggested.

`Candidatus Liberibacter asiaticus´ titers in citrus and acquisition rates by Diaphorina citri are decreased by higher temperature

`Candidatus Liberibacter asiaticus´ is the most prevalent Liberibacter sp. associated with huanglongbing (HLB) in Brazil. Within São Paulo state (SP), HLB has spread more rapidly to and reached higher incidence in regions with relatively mild (cooler) summer temperatures. This suggests that climate can influence disease spread and severity. ?Ca. L. asiaticus? titers on soft, immature leaves from infected ?Valencia? sweet orange plants exposed to different temperature regimes and adult Diaphorina citri fed for 48 h on these plants for ?Ca. L. asiaticus? acquisition were determined by quantitative polymerase chain reaction in two experiments. The first experiment included plants with three levels of infection, three incubation periods (IPs), and air temperatures favorable (14.6 to 28°C) and unfavorable (24 to 38°C) to ?Ca. L. asiaticus?. The second included plants with severe late-stage infections, 10 IPs (based on 3-day intervals over 27 days), and three air temperature regimes (12 to 24, 18 to 30, and 24 to 38°C). Overall, ?Ca. L. asiaticus? titers and the percentages of ?Ca. L. asiaticus?-positive psyllids were lower in plants maintained at the warmer temperature regime (24 to 38°C) than in plants maintained in the cooler regimes. The results suggest that the lower incidence and slower spread of ?Ca. L. asiaticus? to warmer regions of SP are related to the influence of ambient temperatures on titers of ?Ca. L. asiaticus? in leaves.

Elimination of variability sources in NIR spectra for the determination of fat content in olive fruits

Models for prediction of oil content as percentage of dried weight in olive fruits were comput- ed through PLS regression on NIR spectra. Spectral preprocessing was carried out by apply- ing multiplicative signal correction (MSC), Sa vitzky–Golay algorithm, standard normal variate correction (SNV), and detrending (D) to NIR spectra. MSC was the preprocessing technique showing the best performance. Further reduction of variability was performed by applying the Wold method of orthogonal signal correction (OSC). The calibration model achieved a R 2 of 0.93, a SEPc of 1.42, and a RPD of 3.8. The R 2 obtained with the validation set remained 0.93, and the SEPc was 1.41.

Mecanismos implicados en las etapas iniciales de infección en la cancrosis de los cítricos provocada por Xanthomonas citri subsp. citri

La cancrosis o chancro bacteriano de los cítricos (CBC) causada por Xanthomonas citri subsp. citri (Xcc) y X. fuscans subsp. aurantifolii, afecta a un gran número de especies dentro de la familia de las rutáceas, especialmente cítricos. Esta enfermedad produce graves pérdidas económicas allí donde está presente, principalmente porque la comercialización de cítricos desde las zonas afectadas hacía zonas libres de cancrosis, está sujeta a fuertes medidas cuarentenarias. La cancrosis se encuentra distribuida a nivel mundial pero no se ha localizado ni en la Unión Europea ni en ningún área del Mediterráneo. Se han descrito tres tipos de cancrosis en función de la gama de huésped y de las características fenotípicas y genotípicas de las bacterias que las producen. La más extendida es la cancrosis tipo A producida por Xcc, dentro de la cual se distinguen los subtipos Aw y A*, originarios de Florida y Sudeste Asiático, respectivamente, que de forma natural solo son capaces de producir enfermedad en lima mejicana. En este trabajo se presentan estudios sobre mecanismos implicados en las primeras etapas de la infección, como la quimiotaxis y formación de biopelículas, en la cancrosis de los cítricos. La quimiotaxis es el proceso por el cual las bacterias se dirigen hacia zonas favorables para su supervivencia y desarrollo. Los perfiles quimiotácticos obtenidos frente a distintas fuentes de carbono, así como los estudios en relación al contenido de proteínas aceptoras de grupos metilo (MCPs), permitieron agrupar a las cepas de Xanthomonas estudiadas en este trabajo, de acuerdo a la enfermedad producida y a su gama de huésped. Todas las cepas mostraron quimiotaxis positiva frente a extractos de hoja y apoplasto de diferentes especies, sin embargo, Xcc 306, X. alfalfae subsp. citrumelonis (Xac) y X. campestris pv. campestris (Xc) manifestaron respuestas más específicas frente a extractos de apoplasto de hojas de naranjo dulce, lima y col china, respectivamente. Dicho resultado nos permite asociar el mecanismo de quimiotaxis con la capacidad de las cepas de Xanthomonas para colonizar estos huéspedes de forma específica. Las cepas estudiadas fueron capaces de realizar movimiento tipo swimming, twitching y sliding en distintos medios, siendo el movimiento swimming el único en el que se encontraron diferencias entre las cepas de Xcc con distinta gama de huésped. En este trabajo se ha estudiado además la formación de biopelículas en superficies bióticas y abióticas, un mecanismo importante tanto para la supervivencia en superficie vegetal como para el desarrollo de la infección. Las cepas de Xanthomonas estudiadas fueron capaces de formar biopelículas in vitro, siendo mayor en un medio que simula el apoplasto y que contiene una baja concentración de nutrientes en comparación con medios que contenían alta concentración de nutrientes. La formación de biopelículas en superficie vegetal se encontró relacionada, en las cepas patógenas de cítricos, con la capacidad para infectar un tejido o huésped determinado. Se han caracterizado algunos de los componentes de la matriz extracelular producida por Xcc, que compone hasta un 90% de las bipoelículas. Entre ellos destaca el ADN extracelular, que tiene un papel como adhesina en las primeras etapas de formación de biopelículas y estructural en biopelículas maduras. Además, se han identificado el pilus tipo IV como componente importante en las biopelículas, que también participa en motilidad. Finalmente, se han realizado estudios sobre la expresión de genes implicados en motilidad bacteriana y formación de biopelículas que han confirmado las diferencias existentes entre cepas de Xcc de amplia y limitada gama de huésped, así como el papel que juegan elementos como el pilus tipo IV o el flagelo en estos procesos. ABSTRACT Xanthomonas citri subsp. citri (Xcc) and X. fuscans subsp. aurantifolii are the causal agents of Citrus Bacterial Canker (CBC) which is one of the most important citrus diseases. CBC affects all Citrus species as well as other species from Rutaceae family. CBC produces strong economic losses; furthermore the commercialization of plants and fruits is restricted from infested to citrus canker free areas. The disease is worldwide distributed in tropical and subtropical areas, however it is not present in the European Union. Three types of CBC have been described according to the host range and phenotypic and genotypic characteristics. CBC type A caused by Xcc is he widest distributed. Within CBC A type two subtypes Aw and A* were described from Florida and Iran respectively, both infecting only Mexican lime. Herein mechanisms connected to early events in the citrus bacterial canker disease such as chemotaxis and biofilm formation, were studied. Chemotaxis allows bacteria to move towards the more suitable environments for its survival, host colonization and infection. Studies performed on citrus pathogenic Xanthomonas and X. campestris pv. campestris (Xc), a crucifer pathogen, have shown different chemotactic profiles towards carbon compound as well as different MCPs profile, which clustered strains according to host range and disease caused. Every strain showed positive chemotaxis toward leaf extracts and apoplastic fluids from sweet orange, Mexican lime and Chinese cabbage leaves. However, a more specific response was found for strains Xcc 306, X. alfalfae subsp. citrumelonis and Xc towards sweet orange, Mexican lime and Chinese cabbage apoplastic fluids, respectively. These results relate chemotaxis with the higher ability of those strains to specifically colonize their proper host. Xanthomonas strains studied were able to perform swimming, sliding and twitching motilities. The ability to swim was variable among CBC strains and seemed related to host range. Biofilm formation is an important virulence factor for Xcc because it allows a better survival onto the plant surface as well as facilitates the infection process. The studied Xanthomonas strains were able to form biofilm in vitro, on both nutrient rich and apoplast mimicking media, furthermore the biofilm formation by all the strains was higher in the apoplast mimicking media. The ability to form biofilm in planta by Xcc and Xac strains was dependent of the host and the tissue colonized. The wide host range CBC strain was able to form biofilm onto several citrus leaves and fruits, however the limited host range CBC strain produced biofilm solely onto Mexican lime leaves and fruits. Furthermore Xac strain, which solely infects leaves of young plants, was not able to develop biofilms on fruits. Some components of the extracellular matrix produced by Xcc strains have been characterized. Extracellular DNA acted as an adhesin at the very early stages of biofilm formation and as structural component of mature biofilm for citrus pathogenic Xanthomonas. Furthermore type IV pilus has been identified as a component of the extracellular matrix in biofilm and motility. Transcriptional studies of genes related with biofilm formation and motility have confirmed the differential behavior found among wide and limited host range CBC strains as well as the role of type IV pili and flagellum on those processes.

A Gene Encoding a Hevein-Like Protein from Elderberry Fruits Is Homologous to PR-4 and Class V Chitinase Genes1

We isolated SN-HLPf (Sambucus nigra hevein-like fruit protein), a hevein-like chitin-binding protein, from mature elderberry fruits. Cloning of the corresponding gene demonstrated that SN-HLPf is synthesized as a chimeric precursor consisting of an N-terminal chitin-binding domain corresponding to the mature elderberry protein and an unrelated C-terminal domain. Sequence comparisons indicated that the N-terminal domain of this precursor has high sequence similarity with the N-terminal domain of class I PR-4 (pathogenesis-related) proteins, whereas the C terminus is most closely related to that of class V chitinases. On the basis of these sequence homologies the gene encoding SN-HLPf can be considered a hybrid between a PR-4 and a class V chitinase gene.

Optimisation of analytical methods for the characterisation of oranges, clementines and citrus hybrids cultivated in Spain on the basis of their composition in ascorbic acid, citric acid and major sugars

Three HPLC methods were optimised for the determination of citric acid, succinic acid and ascorbic acid using a photodiode array detector and fructose, glucose and sucrose using a refractive index in twenty eight citrus juices. The analysis was completed in <16 min. Two different harvests were taken into account for this study. For the season 2011, ascorbic acid content was comprised between 19.4 and 59 mg vitamin C/100 mL; meanwhile for the season 2012, the content was slightly higher for most of the samples ranging from 33.5 to 85.3 mg vitamin C/100 mL. Moreover, the citric acid content in orange juices ranged between 9.7 and 15.1 g L−1, while for clementines the content was clearly lower (i.e. from 3.5 to 8.4 g L−1). However, clementines showed the highest sucrose content with values near to 6 g/100 mL. Finally, a cluster analysis was applied to establish a classification of the citrus species.