Characterization of the Cell Wall Microdomain Surrounding Plasmodesmata in Apple Fruit.

In fleshy fruits ripening is generally associated with a loss in tissue firmness resulting from depolymerization of wall components and separation of adjacent cells. In the regions of the wall that contain plasmodesmata, the usual sequences of ripening events, i.e. depolymerization of the middle lamellae and splitting of the walls, are not observed. In the present study we attempted to characterize in apple (Malus domestica Borkh.) fruit the structural microdomain of the cell wall that surrounds the plasmodesmata by in muro visualization of the cell wall components. Anionic sites of galacturonic acids were labeled with cationic gold. Low-esterified homogalacturonans were labeled with the monoclonal antibody JIM 5. In addition, a polyclonal antibody directed toward [beta](1->3)-glucopyranose was used to target callose in situ. The results indicated that the plasmodesmata-wall complexes were surrounded by a pectic microdomain. This domain was composed of low-esterified homogalacturonans that were not involved in calcium cross-bridging but were probably surrounded by a cationic environment. These structural features may result in the prevention of normal cell wall separation in regions containing plasmodesmata. However, observations by low-temperature scanning electron microscopy suggested that splitting of these walls ruptured the plasmodesmata and ultimately resulted in the spatial separation of adjacent cells.

Growth of Listeria monocytogenes Restricted by Native Microorganisms and Other Properties of Fresh-Cut Spinach †.

A study was undertaken to investigate the cause of the bacteriostatic activity of fresh-cut spinach leaves against Listeria monocytogenes . L. monocytogenes was cultivated in pure tryptic soy broth for use as a monoculture, in tryptic soy broth containing 10 mg ml-1 of autoclaved or nonautoclaved freeze-dried spinach powder, and in tryptic soy broth in mixed cultures with various microorganisms isolated from fresh-cut spinach, including Pseudomonas fluorescens biovar I, P. fluorescens biovar III, Staphylococcus xylosus , and an undefined culture of mesophilic aerobic microorganisms (MAMs) isolated from freeze-dried spinach powder. These microorganisms were inoculated at 4.4 log CFU ml-1 and L. monocytogenes was inoculated at 2.4 and 4.4 log CFU ml-1 After 24 h of incubation at 30°C, the populations of the two inoculum levels L. monocytogenes increased to 9.0 and 9.6 log CFU ml-1 in the tryptic soy broth control, to 5.4 and 7.5 in nonautoclaved spinach powder cultures, and to 8.8 and 9.1 log CFU ml-1 in autoclaved spinach powder cultures; In mixed cultures with biovar I of P.fluorescens , L. monocytogenes increased to 7.4 and 8.6 log CFU ml-1; with biovar III to 7.7 and 9.1, with S. xylosus to 7.8 and 9.2, and with the MAMs to 7.1 and 8.0 CFU ml-1 in the low and high listerial inoculum cultures respectively. The LSD(0.05)of the means were 0.5 and 0.6, respectively. The freeze-dried spinach powder had an inhibitory effect on the growth of L. monocytogenes . The inhibitory effect was greatly decreased when the native microorganisms were almost eliminated by heating or irradiation. These results indicate that if L. monocytogenes is present as a contaminant on fresh-cut spinach, its growth probably will be restricted by native microorganisms.

Changes in microbial populations on fresh cut spinach.

The microbial populations found on fresh-cut spinach leaves that were stored in gas permeable bags at 10 degrees C for 12 days were examined and identified. The microorganisms consisted of mesophilic aerobic bacteria, psychrotrophic bacteria, Pseudomonadaceae, Enterobacteriaceae, Micrococcaceae, lactic acid bacteria and yeasts. Populations of mesophiles, psychrotrophs, Pseudomonadaceae and Enterobacteriaceae increased sharply during the storage period. The initial populations were 10(7), 10(6), 10(6) and 10(4) CFU.g-1 respectively. Populations reached 10(10) for the mesophiles, psychrotrophs and Pseudomonadaceae and 10(7) CFU.g-1 for Enterobacteriaceae after 12 days of storage. Micrococcaceae, lactic acid bacteria and yeasts remained constant (10(3)-10(4) CFU.g-1. The majority of the bacterial isolates were identified as Pseudomonas fluorescens, Aeromonas caviae and Staphylococcus xylosus. The yeasts, which were most frequently isolated, were classified in the genus Cryptococcus. No pathogens such as Listeria monocytogenes and Salmonella were detected. Observations with low temperature scanning electron microscopy (LTSEM) indicated that the microorganisms were not present on the surface of healthy unbroken leaves. Alternatively, they were found in areas where the cuticle was broken and could be seen infecting the internal palisade parenchyma.

Quantitative high-performance liquid chromatography analysis of plant phospholipids and glycolipids using light-scattering detection.

Application of the evaporative light-scattering principle to quantitative high-performance liquid chromatography (HPLC) analyses of plant membrane lipids has received little study. Light-scattering detection response curves were generated for nine classes of plant membrane phospholipid and glycolipids. Quantitative results obtained by HPLC/light-scattering detection and conventional lipid analytical methods (thin-layer chromatography and lipid-P assay) were in close agreement, confirming the reliability of HPLC/evaporative light-scattering detection (ELSD) analyses. Only three of the nine plant lipid classes gave linear detector response functions above 10 micrograms injected lipid mass. This finding contradicts earlier precepts involving light-scattering detection of lipids. At a given mass, appreciable variation in ELSD signal intensity and detection limit was found to exist among the various plant membrane lipid classes. The variation in detector response among plant lipid classes is an important consideration in achieving accurate quantitative results in plant lipid analyses.

AN ACCURATE, LOW COST FIRMNESS MEASURING INSTRUMENT.

A low-cost, automatic firmness measuring instrument was developed using commercially available components such as: dial gage, dashpot lowering mechanism, weight sets and probe tips. The details of the assembly and the names of parts manufacturers are given. The instrument is an automated modification of the original P-L Meter developed by Parker and Levin at Michigan State University. The application of the instrument to firmness measurements on cherries and tomatoes is summarized and illustrated with typical results. Tomato studies covered the relationship between firmness and variables such as: fruit size, cultivar type, changes in protopectin content, polygalactu-ronase and cellulase activity. The readings compared favorably with those obtained with the Instron Universal Testing machine.

Growth and respiration patterns of snap bean fruits.

The relationship of respiration and growth of seed, pericarp tissue and whole fruit of snap beans (Phaseolus vulgaris L.) was studied. The whole fruit exhibited an apparent climacteric type of respiration pattern. This pattern resulted from an increase in CO(2) production by the enlarging seed followed by a rapid decrease in CO(2) evolution by the pericarp tissue, and the pattern was not associated with any concomitant increase in ethylene production. Therefore, the apparent climacteric respiration pattern of a developing bean fruit is not comparable to the phenomenon that occurs in other ripening fruits.