Characterization of Phaseolus vulgaris L. Landraces Cultivated in Central Italy

Eight Phaseolus vulgaris L. landraces cultivated on farm in marginal areas of Central Italy (Lazio region) were investigated in order to evaluate chemical composition of storage proteins and secondary metabolites fractions. The total protein content showed some differences among landraces; the maximum value was next to 30 g for 100 g of dry weight. The seed storage proteins were screened by polyacrylamide gel electrophoresis (SDS/PAGE): seven landraces exhibited phaseolin patterns type S, one landrace showed a phaseolin pattern type T. A morphological analysis of cotyledon parenchyma performed by scanning electron microscopy (SEM) revealed differences in size of starch granules. Moreover the polyphenolic composition was investigated using HPLC-APCI; from the methanol extracts a flavonoid, kaempferol, and a coumarin, 5,7-dimethoxycoumarin, were identified. To our knowledge, this is the first time that 5,7-dimethoxycoumarin has been reported in P. vulgaris seeds.     

The degradation of lectins,phaseolin and trypsin inhibitors during germination of white kidney beans,Phaseolus vulgaris L.

White kidney beans (Phaseolus vulgaris), cv Processor, contain a relatively high content of phaseolin (storage protein), lectins and a special group of glycoproteins as well as a considerable amount of protein-type trypsin inhibitors. Protein digestion of raw Processor beans in monogastrics, for example pigs, is disturbed by poorly digested, phaseolin lectins, which can bind to carbohydrates in brush border membranes of the small intestinal epithelium, and trypsin inhibitors. The effect of the germination of white kidney beans on lectins, phaseolin and trypsin inhibitors was studied in order to achieve a degradation of lectins, phaseolin and trypsin inhibitors and an increase ofin vitro enzymatic hydrolysis of the protein of bean flour. Therefore, whole bean extracts were examined throughout a germination period of up to seven days for their lectin and phaseolin pattern, lectin content, binding capacities of functional lectins towards brush border membranes and trypsin inhibitor content. In addition thein vitro enzymatic hydrolysis by pepsin and pancreatin of the protein from flours of (un)germinated white kidney beans was studied. SDS-PAGE demonstrated a degradation of E-lectins and a disappearance of L-lectins and phaseolin during germination. Results indicated a decrease of the lectin content by 85%, a loss of binding capacities of functional lectins towards brush border membranes by 91%, and a decrease of trypsin inhibitors by 76%, in bean flour after germination for seven days. A maximum inin vitro enzymatic hydrolysis of protein from bean flour was already established after germination for half a day.     

The use of enzymic hydrolysis in vitro to study the digestibility of some phaseolus seed proteins

In vitro proteolysis was used to study the digestibility of the major seed protein phaseolin, and the trypsin inhibitor, purified fromPhaseolus vulgaris seeds. Whereas denatured phaseolin was fully digested by the enzymic method used [4], its native form was only partially digested. Trypsin inhibitor, both native and after heat treatment, was poorly digested. It was also found that native trypsin had some resistance to digestion conferred upon it by the presence of the inhibitor.     

Denaturation by heat,sodium dodecyl sulphate and dithiothreitol of globulins and phaseolin from dry bean (Phaseolus vulgaris L.)

The electrophoretic pattern of the untreated common bean globulin cv. Flor de Mayo had 7 protein fractions ranging from 620 to 120 kilodaltons (kd). The last molecular weight corresponds to the monomeric form. One of the objectives of the present work was to establish a comparison among denaturation by heat, sodium dodecyl sulphate (SDS) and dithiothreitol (DTT). At pH 6.0, two bands were resistant to heat treatment, after SDS treatment of the untreated globulin several bands disappeared and two new bands with 26 and 15 kd appeared. DTT did not change the electrophoretic pattern, due to the small quantity of free SH groups in the globulin. At pH 6.0, phaseolin is partly resistant to heat denaturation due to aggregation in an acidic environment. Isoelectrofocusing (IEF) and SDS polyacrylamide gel electrophoresis (SDS-PAGE) separated phaseolin into 10 protein fractions. The hypothesis is that phaseolin or globulin aggregation is due to the charge difference of fractions.     

Genetics of intra-gene pool and inter-gene pool hybridization for seed traits in common bean (Phaseolus vulgaris L.) germplasm from Europe

Due to adaptation to new ecological and manmade conditions, the large diversity evolved in the European common bean germplasm is of particular interest for plant breeding. The knowledge of the genetic relationships within and among races and gene pools and their performance per se will provide bean breeders with a starting point in designing crosses using contrasting and complementary parents to broaden the genetic base within the different commercial classes. A genetic study of seed size variation and protein markers in progeny derived from 16 intraracial, interracial and inter-gene pools European common bean populations was conducted. General and specific combining ability (GCA and SCA) values were significant for seed weight, indicating that both additive and nonadditive genetic effects were involved in conditioning seed weight. Interracial populations showed transgressive values due to the accumulation of large-seeded alleles. Genetic variation inside Andean germplasm, and Chile and Peru races in particular, exhibited useful genetic progress in these populations, providing lines with a large seed size, and so, an excellent market potential. The distribution of incompatibility between both gene pools (Mesoamerican and Andean) of the common bean was explored. Inter-gene pool populations provided lower means of inbred segregants than the mid-parent value. Therefore, a good option it would be select for large seed size according to a recurrent or congruity inbred-backcrossing selection programs. Analysis of allele markers frequencies in inter-gene populations showed segregation distortion with a higher than expected frequency of alleles from the Mesoamerican gene pool, many of which were fixed in the F₇ lines. The presence of a great percentage of markers that showed segregation distortion in these populations (87%) indicated that this phenomenon can be amplified by using distance related common bean genotypes. In addition, a high percentage of heterozygotes for the Phs locus (for the seed storage protein phaseolin) was found, which suggest that the Andean homozygous TT could not be expressed in Mesoamerican genetic background due to the action of some form of female specific mechanisms that affected gene exchange between parental germplasm in inter-gene pool populations. The present work provides useful information in the establishment of large seed size germplasm that could have a great deal of interest among breeders and may offer some possibilities to exploit existing variation within and between common bean races.     

Effect of imazamox plus bentazon on dry bean (Phaseolus vulgaris L.)

Imazamox plus bentazon has the potential to provide broad spectrum weed control in dry bean (Phaseolus vulgaris L.). Field trials were conducted in Ontario over a two year period (2003 and 2006) to evaluate the effect of imazamox plus bentazon applied post-emergence at 25 + 600 and 50 + 1200 g ai ha⁻¹ on black, brown, cranberry, kidney, otebo, pinto, white and yellow eye beans. Treatments included a non-treated check. All treatments were maintained weed free during the growing season. The eight market classes responded similarly to imazamox plus bentazon. Imazamox plus bentazon applied post-emergence caused less than 4% visible injury at 7, 14 and 28 days after treatment. Imazamox plus bentazon applied post-emergence did not have any effect on plant height, shoot dry weight, seed moisture content and yield of dry bean. Based on these results, imazamox plus bentazon at the proposed maximum use dose can be safely used for weed management in black, brown, cranberry, kidney, otebo, pinto, white and yellow eye bean production in Ontario. Additional research is needed to determine if cultivars within a market class of dry bean differ in their response to imazamox plus bentazon.     

Methionine-containing proteins in twoPhaseolus vulgaris cultivars with different methionine bioavailabilities

The full utilization of legumes as human food is limited by a deficiency of sulfur amino acids, low protein digestibility, low methionine bioavailability and the presence of anti-nutritional factors. A new cultivar ofPhaseolus vulgaris (Carioca 80) has 56.8% available methionine, compared with 29.3% availability in the parent cultivar Carioca. The total methionine content, denaturing gel electrophoretic patterns of methionine-containing proteins, and the percentage of phaseolins (the major storage proteins in Phaseolus) relative to the total protein are similar in the two cultivars. Although the digestibility of the two cultivars is similar, the increased biological value of Carioca 80 may indicate that there are differences in overall bean composition that affect protein hydrolysis and utilization. We suggest the tentative explanation that this is due to differences in the distribution of methionine in the methionine-containing proteins of the two cultivars.     

Biomass distribution,maturity acceleration and yield in drought-stressed common bean cultivars

Common bean (Phaseolus vulgaris L.) is an important food crop grown under rainfed conditions in Latin America where drought is a major limiting factor for production. The objective of this study was to assess the role of phenological adjustment and shoot biomass distribution on seed yield of drought-stressed common bean. Four cultivars differing in growth habit, gene pool origin, and contrasting responses under drought, were tested during 2001 at two locations in Mexico: Cotaxtla, Veracruz (lowlands) where the effect of terminal drought (TD) (end-of-season) was evaluated, and Texcoco, State of Mexico (highlands), where the effects of intermittent and terminal drought were evaluated. Seed yield, plant shoot biomass, and days to flowering and to physiological maturity were recorded. Leaf relative water content (RWC) was recorded after the onset of the intermittent drought (ID) treatment in Texcoco. The drought intensity index was 0.37 in Cotaxtla compared to 0.49 and 0.58 under TD and ID, respectively in Texcoco. Days to flowering and to physiological maturity showed a negative and significant relationship with seed yield. Under drought stress, a significant reduction in the harvest index was observed in susceptible cultivars. All cultivars showed higher values of shoot biomass accumulation, pod and seed number, seed weight and RWC at the basal nodes of the plant across locations and moisture treatments. Cultivar Pinto Villa exhibited the highest biomass accumulation and seed yield across treatments and locations. Significant reduction in number of days to maturity was observed under drought, mainly in resistant cultivars, Pinto Villa and G4523. Maturity acceleration, coupled with a high seed filling rate, contributed to lessen the impact of drought stress in resistant common bean cultivars.     

Nutritional potential ofVigna minima (Roxb.) Ohwi and Ohashi:

Quantitative variation in different fractions of seed proteins and their amino acid levels in populations ofVigna minima (Roxb.) Ohwi and Ohashi and inV. umbellata cv. IC 1568 — the rice bean — were investigated. Globulin I fraction, together with globulin II, constitutes 38 to 54 per cent of the total seed protein. The alkali soluble (glutelin) fraction is the second largest fraction. Both these fractions show broad range of variation, suggesting a broad genetic base. The profiles are population specific; the coastal population, which contains higher seed protein also possesses maximum levels of globulin I and glutelin fraction suggesting its potentiality for breeding lines with high protein content, high nutritive value, and salt tolerance. Protein content is positively correlated with globulin I and glutelin fractions, which are in turn positively correlated with each other. The amino acid profiles are specific not only to the fractions but also to the populations. The range of variation in the levels of all amino acids in different fractions is broad suggesting substantial genetic diversity. The average levels of lysine and sulphur amino acids are high in globulin I and glutelin fractions.     

Divergent selection at ultra-low plant density for seed protein and oil content within soybean cultivars

Soybean [Glycine max (L.) Merr.] seed is a major source of protein for animal feed and oil for human consumption. Selection within elite soybean cultivars for the improvement of agronomic and seed traits is assumed to be ineffective due to the belief that cultivars are highly homogeneous. Previously reported data suggest that latent variation among the single plant selections within a cultivar exists and that mechanisms that generate de novo variation may also be present. The main objective of this study was to perform divergent single-plant selection at ultra-low plant density and investigate the presence of genetic variation for seed protein and oil within three elite soybean cultivars. A secondary objective was to investigate the variation for fatty acid composition. In 1995, single plants from the three cultivars were grown in a honeycomb design using a plant-to-plant spacing of 0.9 m. A total of 333 plants from ‘Benning’, 392 plants from ‘Haskell’, and 371 plants from ‘Cook’ were evaluated. Divergent single-plant selection for protein and oil content was performed to select a total of 20 plants for high or low protein and 20 plants for high or low oil from each cultivar. The selected plants were further evaluated in replicated row-plot experiments for 3 years. Our results indicate that single-plant selection at low plant density was successful in discovering significant variation for seed protein and oil within each of the three soybean cultivars. For protein content, the magnitude of intra-cultivar variation between the highest- and lowest-protein lines averaged 19 g kg⁻¹ across the three cultivars and ranged from 13 to 24 g kg⁻¹. For oil composition, the magnitude of variation between the most divergent lines averaged 12 g kg⁻¹ across the three cultivars and ranged from 9 to 14 g kg⁻¹. Significant variation among the selected progeny lines was also discovered for specific fatty acid composition. The magnitude of intra-cultivar variation averaged from 6 to 29 g kg⁻¹ across the five fatty acids of soybean. The genetic variation discovered within the soybean cultivars is most likely due to latent variation and/or newly created variation. Our data provide evidence that single-plant selection at ultra-low plant density within elite cultivars can be effective in improving the seed composition of a soybean cultivar.     

Identification of sources of resistance to damping-off and early root/hypocotyl damage from Rhizoctonia solani in common bean (Phaseolus vulgaris L.)

Rhizoctonia solani causes economically important root and hypocotyl diseases in common bean throughout the world. Root health is a vital factor in plant development and root diseases would negatively influence water and nutrient uptake as well as cause direct stand reduction and root rot damage to the crop. An efficient common bean screening method to evaluate damping-off and early root/hypocotyl damage from R. solani was developed and used to identify dry bean lines with levels of resistance to this disease. Two sets of 163 and 111 lines previously evaluated for drought tolerance in Nebraska and Puerto Rico were evaluated for damping-off resistance and early root/hypocotyl damage under greenhouse conditions. Disease severity on plants was identified based on above-ground symptoms, seedling survival and root lesions using a rating scale of 1 (resistant) to 9 (susceptible). In the first set of lines representing commonly grown dry bean cultivars, germplasm and sources of damping-off resistance, the Rhizoctonia mean rating ranged from 1.7 to 3.9; Phaseolus vulgaris lines PI 310668 and PI 533249 had the highest damping-off resistance. In the second set of the best lines from a drought tolerance shuttle breeding program the Rhizoctonia mean rating was between 2.6 and 5.7. The availability of drought tolerant dry bean lines allowed the testing of the hypothesis that there was a correlation between selecting for drought tolerance and R. solani damping-off resistance. No correlation between mean disease rating and drought tolerance was found, but adapted dry bean lines such as NE14-08-176 released as SB-DT1, and NE14-08-225 were identified with moderate damping-off resistance and drought tolerance. Lines with both traits and other attributes will facilitate development of resistant bean cultivars to manage damping-off caused by R. solani.     

Seed Weight of Nodulating and Non-nodulating Soybeans at Different Nitrogen Levels and Years

Abstract

Seed weight (weight per seed) is an important trait in soybean [Glycine max (L) Merr.] that affects its production, processing, marketing and consumer preferences. To determine the effects of nitrogen fertilizers and climatic conditions on seed weight, a five-year field experiment was conducted by growing four nodulating cultivars and 13 non-nodulating lines at 4 levels of nitrogen fertilizer (0, 2, 10 and 20 g m^–2). The variation in seed weight due to the difference in the fertilizer level and climatic condition of the year was greater in non-nodulating lines than in nodulating cultivars. This resulted in a lower heritability estimate in non-nodulating lines (0.49) than in the nodulating cultivars (0.85). Nitrogen fertilizer increased the seed weights of non-nodulating lines but did not affect the seed weights of the nodulating cultivars. The high response of the non-nodulating lines to nitrogen fertilizer may be attributed to their strong dependence on fertilizer and soil nitrogen due to their lack of ability to fix nitrogen in symbiosis. The small seeds produced during the years with low temperatures and early onset of frost may have been caused by the slow seed growth rate and short seed filling duration.     

Preparation and antinutritional characteristics of dry bean (Phaseolus vulgaris L.) protein concentrates

Protein concentrates and starches were prepared by a wet extraction process from five dry bean (Phaseolus vulgaris L.) cultivars. The protein contents ranged from 69.7–76.4%. Concentrates prepared from dehulled beans under similar conditions had higher protein contents (80.6–87.9%). Each additional washing of the concentrates with distilled water increased their protein content. However, the protein recovery progressively decreased. The yield of starch ranged from 48.0–51.1% of the starting material. The solubility of bean proteins was minimal at pH 4.0, and under alkaline conditions, it was influenced by the tannin contents of the concentrates. Protein concentrates had lower trypsin, chymotrypsin, and amylase inhibitory activities as well as lower phytic acid and tannin contents compared to whole bean flours.     

Tannins,Trypsin Inhibitors and Lectin Cytotoxicity in Tepary (<Emphasis Type="Italic">Phaseolus acutifolius</Emphasis>) and Common (<Emphasis Type="Italic">Phaseolus vulgaris</Emphasis>) Beans

This study compared the levels of antinutritional components and cytotoxic effect of extracts, from tepary (Phaseolus acutifolius) and common (Phaseolus vulgaris) beans. Antinutritional factors were evaluated by determining their effect on the viability of epithelial cells isolated from rat small intestine. The protein and carbohydrates content were similar in all the genotypes studied (20 and 60%, respectively). Common beans presented higher content of trypsin inhibitors, tannins and lectins than tepary beans. There was not a significant correlation between tannins and cooking time. However, water absorption and cooking time correlated significantly (p < 0.05). Considerable variation was observed in lectin activity (1302–18161 Ul/mg) of extracts from different beans. Tannins, lectins, trypsin inhibitors and fat content differed between bean varieties whereas protein content was similar. The percent cellularity on rat epithelial cells was significantly different among protein extracts from different bean cultivars and ranged between 53.5% and 87.4% (p < 0.05). These results suggest that the incorporation of tepary beans in the diet would not alter the current nutritional contribution of common beans or introduce adverse toxic effects. The agronomic characteristics of tepary beans make them attractive for cultivation. However, the harder to cook phenomenon may be a limiting factor that needs further consideration.     

Variability of Nutritional and Cooking Quality in Bean (Phaseolus vulgaris L) as a Function of Genotype

Screening of natural biodiversity for the better quality traits are of prime importance for quality breeding programs. The objective of this investigation was to select candidate accession of bean having high concentrations of protein as well as macro and micro minerals with good cooking quality for use as parents in breeding programme for these compounds. Thirty-five accessions of bean (Phaseolus vulgaris L) were field grown and their seeds were analyzed for their cooking quality and nutritional composition. Wide variations were observed in most of the measurements e.g. protein (18.7–26.2%), iron (79.4–137.6 ppm) and hardness after cooking (4.65–9.88 Kg) suggesting that there are considerable levels of genetic diversity. Across all accessions the concentration of potassium was negatively correlated with protein (r = −0.43, P < 0.05). Concentrations of protein was significantly greater in accessions VIII, XIII and XIX compared to other accessions analyzed. Iron concentrations were greatest (137 ppm) in XIX and lowest (79 ppm) in XXVII. Lines with less cooking time were line III, X, XXVI, XXX and XXXI. Bean line XIX contains high protein (24.9%) with high zinc (33.3 ppm) and highest iron (137.6 ppm), but it has high hardness after cooking (7.32 kg). Four clusters were computed by cluster analysis that explained quite a good variation in the traits. The great variability for these attributes suggests that these selected accessions may be useful as parents in hybridization programs to produce bean with value-added traits. This information was also potentially useful for pulse breeders working on the development of new varieties.     

IgE-Mediated Cross-Reactivity among Leguminous Seed Proteins in Peanut Allergic Children

The immunological cross-reactivity among major protein- and oil-crops, including lupin, lentil, pea, peanut, kidney bean and soybean, has been studied by a combination of in vitro and in vivo experimental approaches: SDS-PAGE separations of legume protein extracts and immuno-blot revelations with 12 peanut-sensitive subjects’ sera, Immuno-CAP and Skin Prick tests on the same subjects. The immuno-blotting data showed a wide range of IgE-binding responses both displayed by one subject towards different plant extracts and among subjects. Differences were both quantitative and qualitative. The prevalent responses of most subjects’ sera were seen with peanut polypeptides, as expected, as well as with various polypeptides of the other legumes, the most recurrent of which were the basic subunits of the 11S globulins. The distribution of in vivo responses generally paralleled those obtained by in vitro approaches with strong responses elicited by peanut, lentil and pea protein extracts, especially by most sensitive subjects, thus providing a consistent overall set of results. In this work, the comparison of various approaches has allowed us to get an overall broad picture of the immunological cross-reactivities among proteins of widely used different seed species and to hypothesize the role of most conserved specific polypeptides.     

Aluminum contents of the edible portions of the winged bean,Psophocarpus tetragonolobus (L.) DC. (Fabaceae): Field study and caveat

The winged bean,Psophocarpus tetragonolobus (L.) DC has been widely recognised as a potentially useful source of protein for tropical regions of the world. A neglected aspect of the plants nutritional quality has been the reporting of the accumulation of aluminum (Al) in the edible parts of the plant; the leaves, pods, seeds and tubers. The accumulation of Al in the plant on acidic tropical soils and the implication of Al to human health problems further justified the inclusion of Al in nutritional analysis. Field experiments on two varieties of the winged bean, the USDA-releaseHi-Flyer and an unnamed variety from the Phillipines, showed that all edible portions of the plant accumulate Al from high to very high levels when compared to an average of usually less than 300 ppm in other crop plants. Aluminum accumulation is generally highest in the youngest tissues particularly in the young roots with levels recorded as high as 25,000 ppm and these contents are as high as levels for recognised Al accumulators such as the leaves ofPinus and tea. Future improvement breeding programs for the winged bean will necessarily require identifying and taking advantage of possible variation in Al accumulation between varieties.     

Interrelationships between certain physical and chemical properties of dry bean (Phaseolus vulgaris L.)

The interrelationships among certain physical and chemical properties of ten cultivars of dry beans (Phaseolus vulgaris L.) were investigated. The length/breadth ratio was similar (1.51–1.65) except for kidney beans (>2.0) and sanilac (1.37) cultivars. The breadth/thickness ratio ranged from 1.17–1.65. The 100-bean weight indicated a wide variation of 15.03–50.33 g. The density, bulk density, and porosity characteristics were within a narrow range of 1.18–1.36 g/cc, 68–75 g/100 cc, and 40.7–48.5%, respectively. Water uptake rates during the first 6 h of soaking at room temperature (21 °C) were characteristic of the cultivar. At the end of 24-h soaking, however, all cultivars had absorbed similar amounts of water (approximately 1 g/g bean). Leaching losses (g solids leached/100 g beans) had characteristic trends and ranged from 0.54 for cranberry to 3.46 for sanilac cultivars after 24-h soaking. Most correlations between selected chemical constituents and physical characteristics of the dry bean were relatively low.Journal Paper No. 2751 of Utah Agricultural Experiment Station and a contribution of Western Regional Project W-150.     

The Null-4A Allele at the Waxy Locus in Durum Wheat affects Pasta Cooking Quality

Granule-bound starch synthase, also known as the waxy protein catalyses the synthesis of amylose in wheat endosperm starch. In durum wheats, the genes encoding GBSS are present at the two Wx loci on chromosome 7A and 4A (a segment of 7B that has been translocated). Several null Wx-B1 (missing GBSS protein from chromosome 4A) durum lines were produced from crosses with null-4A bread wheats backcrossed to durum wheats. Semolina milled from 4 normal and 7 null-4A durum wheat lines grown over two seasons (1999 and 2000) in South Australia were analysed for amylose content, starch pasting properties as measured by the Rapid Viscoanalyzer (RVA), swelling power and starch damage, protein content and electrophoretic protein analysis. Spaghetti was prepared with a micro-scale extruder and the cooked pasta evaluated for cooking loss, firmness, stickiness and water absorption. The null-4A lines had significantly lower (ca. 5%) amylose content, higher starch peak viscosities and semolina swelling power. The pasta derived from the null-4A lines had lower cooking loss and in 1999 was more adhesive than the non-waxy lines. Cooking loss was correlated with amylose content, peak starch viscosity, swelling power of semolina and cooked pasta adhesiveness. Semolina swelling power was highly correlated with RVA peak viscosity. Waxy durum wheats appear to have an advantage over the normal types in terms of lower cooking loss, widely used as an indicator of pasta cooking quality.     

Variation in parameters related to leaf thickness in common bean (Phaseolus vulgaris L.)

Greater leaf thickness is usually associated with greater leaf carbon assimilation rate, but the tradeoff between thicker versus larger leaves also affects canopy structure and light interception. Many studies equate the ratio of dry leaf mass to area (leaf specific mass) with leaf thickness. This approximation has utility but ignores differences in true thickness due to variation in water or air content and other factors. To understand better variation in leaf thickness in common bean (Phaseolus vulgaris L.), 14 parameters related to thickness were compared for a diverse set of bean cultivars grown in three field environments in Colombia. The parameters included leaf thickness per se as measured anatomically or with a micrometer, leaf specific mass expressed on dry and fresh weight bases, leaf optical density measured at 670 nm, total leaf chlorophyll concentration expressed on a leaf area basis, leaf tissue density, and nitrogen concentration expressed on fresh and dry weight bases. Relative contributions of air, water, and dry matter to leaf thickness were also estimated. Most parameters showed large differences among lines, consistent with previous reports that cultivars from the Mesoamerican genepool have thicker leaves than those of the Andean genepool. Parameters varied greatly with environment and sampling date. The results support the need to control for leaf water content when studying leaf thickness. Furthermore, including mid and lateral rib tissue in leaf samples appeared to bias data sufficiently enough to double estimates of leaf thickness and specific mass. Based on time required for measurement and ability to detect cultivar differences, measuring leaf thickness with a micrometer and leaf optical density appeared the most promising for rapid characterization of leaf thickness.