A mechanics model for the compression of plant and vegetative tissues

The mechanics analysis of plant or vegetable tissue under a compressive stress has been developed based on large deformation elasticity theory. The tissue was treated as a lattice of regular perfect three-dimensional hexagonal cells. The cell walls were assumed to be impermeable under the time-scale of the loading. The cell walls of plants and vegetables are polymeric composite materials, consisting of a relatively amorphous matrix and a highly structured network of microfibrils embedded in the cell wall matrix. The micromechanical features of the individual cells have been related to the macroscopic properties of the whole tissue. The effects of microfibrillar stiffening factors k(1) and k(2), the cell wall matrix property alpha and the initial cell expansion ratio nu(i) on the compressive behaviour of a plant or vegetable tissue have been investigated. The predicted results have also been related to some experimental evidence.     

A sensitive method for determining quinolinic acid in animal tissues

A sensitive chromatographic method for isolation and measurement of quinolinic acid from rat liver and kidney is described. The method is based on the isolation of quinolinic acid by ion-exchange chromatography. The extraction of quinolinic acid consisted of the freeze clamping of the organ in liquid nitrogen, followed by deproteinization in perchloric acid. The neutralized extract was concentrated by freeze-drying and submitted to the action of concentrated perchloric acid to hydrolyze the nucleotides which interfered in the chromatographic separation of quinolinic acid. The sample was applied to a column of Dowex (HCOO^?) and eluted with a linear gradient of formic acid. The eluted fraction containing quinolinic acid was quantitatively measured by its absorbance at pH 2 and 268 nm in a spectrophotometer.     

A quantitative cytochemical method for phosphofructokinase in plant tissues

A quantitative cytochemical method for the demonstration of phosphofructokinase has been successfully applied to a range of plant tissues. The findings indicate that this enzyme system may be assayed as an indicator of glycolytic activity in plant cells, and furthermore tha the very high endogenous phosphoenolpyruvate concentrations may not be rate limiting in vivo.     

Digital photography provides a fast,reliable, and noninvasive method to estimate anthocyanin pigment concentration in reproductive and vegetative plant tissues

Anthocyanin pigments have become a model trait for evolutionary ecology as they often provide adaptive benefits for plants. Anthocyanins have been traditionally quantified biochemically or more recently using spectral reflectance. However, both methods require destructive sampling and can be labor intensive and challenging with small samples. Recent advances in digital photography and image processing make it the method of choice for measuring color in the wild. Here, we use digital images as a quick, noninvasive method to estimate relative anthocyanin concentrations in species exhibiting color variation. Using a consumer‐level digital camera and a free image processing toolbox, we extracted RGB values from digital images to generate color indices. We tested petals, stems, pedicels, and calyces of six species, which contain different types of anthocyanin pigments and exhibit different pigmentation patterns. Color indices were assessed by their correlation to biochemically determined anthocyanin concentrations. For comparison, we also calculated color indices from spectral reflectance and tested the correlation with anthocyanin concentration. Indices perform differently depending on the nature of the color variation. For both digital images and spectral reflectance, the most accurate estimates of anthocyanin concentration emerge from anthocyanin content‐chroma ratio, anthocyanin content‐chroma basic, and strength of green indices. Color indices derived from both digital images and spectral reflectance strongly correlate with biochemically determined anthocyanin concentration; however, the estimates from digital images performed better than spectral reflectance in terms of r² and normalized root‐mean‐square error. This was particularly noticeable in a species with striped petals, but in the case of striped calyces, both methods showed a comparable relationship with anthocyanin concentration. Using digital images brings new opportunities to accurately quantify the anthocyanin concentrations in both floral and vegetative tissues. This method is efficient, completely noninvasive, applicable to both uniform and patterned color, and works with samples of any size.     

Spectrophotometric method for determining the stability of lysosomes from animal tissues depending on the hydrogen ion concentration

Rapid spectrophotometric method of lysosome stability determination depending on hydrogen ion concentration is described. The time of analysis is decreased by 5-6 h in comparison with enzymic method. The process of lysosome degradation was linear at pH 6. The incubation mixture acidity dependence curve of lysosome lysis extend was complex. The lysosome lysis rate rapidly increased at pH much less than 6 less than pH. Lysosome incubation at 0-4 degrees C during 24 h decreased its sensitivity to incubation mixture acidity within the whole investigated pH range. Isolated lysosome acid resistance may be used as an index of its stability and lability in vivo and in vitro by various physicochemical factors. Percentage of initial absorbtion (A520) and initial lysosome lysis rate (delta A520/min) may be index of such effect.     

Rates of production and internal levels of ethylene in the vegetative cotton plant

The internal levels of ethylene in and the production ratesof ethylene by various parts of the cotton plant {Gossypiumhirsutum L. cultivar Acala 4–42) were determined by separatetechniques. Wounding of tissue, which resulted from both techniques,increased ethylene production. A peak in wound-induced ethyleneoccurred around the second to third hour after wounding andgradually declined until the sixth or seventh hour when ethyleneproduction began to level off. Using two techniques, with numerousobservations from each technique, ethylene levels were foundto be relatively uniform throughout the vegetative cotton plant.There was a small trend for levels of ethylene to increase fromthe bottom (oldest tissue) to the top of the plant (youngesttissue). Of major interest was the observation that the productionrates and internal concentrations of ethylene in the petiolewere two to six times higher than those in the leaf blade. Thisrelative concentration of ethylene in the petiole supports thehypothesis that ethylene is a natural regulator of abscissionwhich acts, in part, through modification of auxin transport. ¹ A contribution of die Texas Agricultural Experiment Station.Supported in part by Grant GB-5640, National Science Foundationand grants from the Cotton Producers Institute. The data forthis paper was taken from a Master of Science Thesis by J. A.M. submitted to Texas A & M University January 1970. ² Present address: Department of Agronomy, Purdue University,Lafayette, Indiana, U.S.A. (Received June 29, 1971; )     

A new method for determining the minimum inhibitory concentration of essential oils

A new microdilution method has been developed for determining the minimum inhibitory concentration (MIC) of oil-based compounds. The redox dye resazurin was used to determine the MIC of a sample of the essential oil of Melaleuca alternifolia (tea tree) for a range of Gram-positive and -negative bacteria. Use of 0·15% (w/v) agar as a stabilizer overcame the problem of adequate contact between the oil and the test bacteria and obviated the need to employ a chemical emulsifier. A rapid version of the assay was also developed for use as a screening method. A comparison of visual and photometric reading of the microtitre plates showed that results could be assessed without instrumentation; moreover, if the rapid assay format was used, rigorous asepsis was not necessary. Accuracy of the resazurin method was confirmed by plate counting from microwells and MIC values were compared with results obtained using an agar dilution assay. The MIC results obtained by the resazurin method were slightly lower than those obtained by agar dilution.     

A standardized method for determining buffering capacity of plant foliage

Summary A method for determining the buffering capacity (B.C.) of foliage extracts was standardized and evaluated. Sources of variations (biological, field and laboratory) were identified. These variations were reflected in inter-specific differences, seasonal fluctuations, age of the foliage and duration and the conditions of storage of the extracts. Procedures have been recommended to eliminate or minimize sources of variations (other than inherent specific) by standardizing the field sampling, laboratory processing and methods, and calculations of the buffering capacity. Plants such as lichens, known to be sensitive to air pollutants, had very low B.C. whereas species of intermediate sensitivity such as balsam fir had higher B.C. The B.C. being inherited and significantly different among species, has potential for its use in indexing the relative sensitivity of species to air pollutants especially in areas where large numbers of species are to be compared.     

A solid phase method of determining DNA sequence

A solid-phase method for DNA sequencing has been developed which involves immobilization of the terminally labeled DNA fragment on the DEAE-paper followed by chemical modification and cleavage at G, A + G, C + T, and C sites. As compared to the Maxam and Gilbert method, the new technique is more rapid and less laborious, being of the same efficiency.     

A reliable method for the estimation of DNA in higher plant tissues.

A method for the isolation of the calcinogenic principle of Solanum malacoxylon is described. The procedure includes preliminary purification of an aqueous leaf extract by dialysis and treatment with ion-exchange resins. Final purification is achieved on Sephadex G-15 and G-10 columns and by paper chromatography.     

A new method for determining cross-sectional shape and area of soft tissues

Assessment of the mechanical properties of soft tissues requires accurate measurement of the cross-sectional area. To date, techniques for determining cross-sectional areas of ligaments and tendons have been less than ideal due to the tissues' complex geometries and the fact that they deform easily under an applied external load. A new procedure has been developed for determining the cross-sectional area by means of an image reconstruction technique based on measurements from collimated laser beams. Using this procedure, the actual shape of the specimen cross-section can also be determined. The results are demonstrated to be highly accurate, and this methodology does not require mechanical contact with the specimen.     

A rapid spectrophotometric method of determining heat-resistant lysosomes in animal tissues

Rapid spectrophotometric method for the determination of thermoresistance in tissue animal lysosomes is described. The of analysis is decreased by 5-6 h, in comparison with enzymatic technique. The determination regimen was chosen in such a way that the process of lysosomal lysis was linear. The dependence of the incubation mixture temperature on the degree of lysosomal lysis was complex. The rate of lysosomal lysis rapidly increased at greater than 37 degrees C. Lysosome incubation at 0-4 degrees C for 24 h decreased its hypothermal (t = 10-30 degrees C), but not hyperthermal (t greater than 37 degrees C) sensitivity. Isolated lysosome thermoresistance may be used as an index of its stability and labialization in vivo and in vitro by various physico-chemical factors. The percentage of initial absorption (A520) and the initial rate of lysosomal lysis (delta A520/min), as well as melting temperature (Tmel) and biological half-life (t1/2) may be the measurements of such effect.     

A simplified method for determining 1-aminocyclopropane-1-carboxylic acid (ACC) in plant tissues using a mass selective detector

A sensitive and specific method is described for the routine assay of the ethylene precursor 1-aminocyclopropane-1-carboxylic acid (ACC) in 100–200 mg fresh weight samples of green or etiolated tissue. The method involves high performance liquid chromatography (HPLC) and gas chromatography linked to mass spectrometry (GCMS) and uses ¹⁴C-labelled ACC as an internal standard, N-benzoyl n-propyl ACC as an easily prepared derivative for HPLC and GCMS, and N-benzoyl isobutyl ACC as an internal standard for GCMS. The procedure is faster and safer than an existing GCMS method and more specific and reliable than indirect assays widely in use. The method has been used to measure ACC in maize roots, young leaves of cucumber, and aerobic or anaerobic seedlings of rice.