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Jose Lopez-Medina, B.J. Murphy, and J.N. Moore

Isozyme staining and SDS-PAGE of soluble proteins were performed using leaf homogenates from 6- to 8-month-old field-grown seedlings resulting from the cross of either `Heritage' or `Nova' raspberry with ARK-577 diploid blackberry, the latter used as the pollen-donor parent. Four enzyme systems were tested: ADH, PGM, MDH, and PGI. Of these, ADH and PGM did not show any activity; MDH was monomorphic in both raspberry and blackberry parents, with activity at the same migration distance. PGI was polymorphic in the two raspberry cultivars, showing three dimeric bands, but monomorphic in blackberry; the allele for PGI in blackberry being common to that allele coding for the most cathodal band in raspberry. This phenomenon, in addition to poor resolution of bands (due, perhaps to low enzyme activity) and evidence of accidental self-pollination in our populations, prevented us from positively identifying the hybrid offspring using isozymes. By SDS-PAGE of soluble proteins, two bands were detected that seemed to be unique to ARK-577 blackberry and were also expressed in some of the offspring, suggesting a hybrid origin of these seedlings. Morphological comparisons indicated that those seedlings possessing the two unique bands highly resemble the blackberry male parent, greatly supporting their hybrid origin. Unless additional analysis shows otherwise, SDS-PAGE can be used to identify Raspberry–Blackberry hybrids during their vegetative stage of development, and might prove applicable to other interspecific hybrids of Rubus.

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J.A. Kirkpatrick, J.B. Murphy, and T.E. Morelock

Interest in the health benefits of vegetables prompted an investigation of the levels of carotenoids in commercial varieties and UA breeding lines of spinach. Plant carotenoids perform a critical function as antioxidants, providing protection against a variety of reactive oxygen species generated primarily during photosynthesis. When ingested by humans, these compounds maintain their antioxidant activities and are receiving considerable attention in relation to multiple health benefits, including cancer prevention. While the best-known and most-studied carotenoid is beta-carotene, other carotenoids are now receiving attention due to their higher antioxidant activity compared to beta-carotene. Most dark-green leafy vegetables, such as spinach and kale, are relatively high in carotenoids, especially lutein. In this study, significant differences in average content of both lutein and beta-carotene were found between genetic lines of spinach. Some lines exhibited considerable variation between plants, while others were highly uniform. There was a very high correlation (r 2 = 0.96) between lutein content and betacarotene content. The significant difference between spinach lines suggests that improvement of general carotenoid antioxidants and lutein could be obtained through a breeding program.

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Maria Elena Garcia, C.R. Rom, and J.B. Murphy

The level of dihydroxyphenolics as influenced by cultivar and leaf phenology was determined for 10 apple cultivars. Newly unfolding leaves were tagged weekly. Once per month, the leaves were collected and analyzed to determine their phenolic content. Replicated collections were made at three times during the summer. Diphenyl boric acid ethanol was the reagent used in this spectrophotometric technique. The phenolic content changed throughout the season. However, no trend was observed for phenolic content variation with leaf age. Each cultivar varied independently in phenolic content with leaf phenology. There were variations in the phenolic content associated with cultivar. Possible hypotheses as to factors affecting the production of phenolics in apple are discussed.

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M.E. Garcia, C.R. Rom, and J.B. Murphy

The effects of shading and leaf age on the production of foliar phenolics of two apple (Malus domestica Borkh.) cultivars, `Liberty' and `Red Rome Beauty', were studied. Potted trees were grown outdoors and their leaves tagged weekly when they reached 20 mm in length. This process continued for the duration of the experiment. At 3 weeks from budbreak, the trees were placed in three shade treatments: 0% shade (control), 60% shade, and 90% shade. After 5 weeks, the leaves were collected for phenolic assay. Specific leaf weight (SLW) was determined from the leaf below the tagged leaf. Shade significantly affected the total phenolic content. Leaves in 0% shade had the highest levels of total phenolics. The phenolic content decreased with increasing shade, with trees in 90% shade having a 72% reduction in total phenolics. There was a significant shade by leaf age interaction. There was a decrease in total phenolic content with increasing leaf age except for those leaves whose development occurred before the experiment was started. The 1-week-old leaf had the highest phenolic content, while 4-week-old leaf had the lowest amount. The 5- and 6-week-old leaves that had been tagged prior to the onset of the shade treatments has similar phenolic content in all treatment. SLW significantly decreased with increasing shade and increased with leaf age. Results of this study indicate that light and leaf developmental stage are important factors in the total foliar phenolic content, but, once phenolics are synthesized, shading does not affect their content.

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Hallie G. Dodson, J.B. Murphy, and T.E. Morelock

Anthocyanins are naturally occurring plant pigments that are classified as flavonoids. Anthocyanins have important antioxidant properties which may help in prevention of cancer, arthritis, and cardiovascular disease. Finding common sources and possibly increasing levels of anthocyanins in food could be important to human health. This research project determined amount and type of anthocyanins in 16 cultivars and breeding lines of cowpeas (Vigna unguiculata). The information obtained from this research project will be used to improve anthocyanin content of cowpeas by breeding. Of the 16 cultivars and breeding lines, only a black cowpea breeding line, 95-356, contained measurable levels of 3 types of anthocyanins: delphinidin, peonidin, and an unknown anthocyanin. Total anthocyanin content was 0.00242 mg·g–1, which is equal to 0.21 mg per ½-cup serving. In another study with 95-356 Rabi A. Musah, found a total anthocyanin content of 121.26 mg per serving and also found three other types of anthocyanins. The difference in the studies could be explained by the storage time of two weeks after the anthocyanins were extracted, but before they were eluded in this study. Additional studies are needed to determine if cowpeas can provide anthocyanin levels comparable to other fruits and vegetables.

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M. Elena Garcia, C.R. Rom, and J.B. Murphy

Two experiments were conducted to determine the effects of leaf age and shading on the phenolic content and composition of apple foliage. In the first study, it was determined that the phenolic content of `Liberty', at increasing leaf developmental stages, was leaf age—dependent. Early during leaf development, there was an increase in the phloridzin (the primary glycoside identified) and in total phenolics, reaching a maximum when the leaf is 6 days from 20-mm blade length. After this stage, the phenolic content decreased with increasing leaf age. In the second study, the leaves of two cultivars, `Liberty' and `Starkspur Law Rome', were tagged weekly when the leaf was two-thirds unfolded. Three weeks after budbreak, the trees were placed under three shade cloth treatments (0%, 60%, and 90% shade). After 4 weeks under the shade treatments, the tagged leaves were collected to determine their phenolic content. Shade significantly affected the foliar phenolic content. Leaves in 0% shade had the highest phenolic content, whereas the lowest content was found in leaves exposed to 90% shade. There was a significant leaf age × shade interaction. The phenolic content decreased with increasing leaf age except for those leaves whose development occurred before the experiment was started. The results indicate that light and leaf developmental stage are important factors in determining the phenolic content of apple leaves, but shading appears to have a stronger influence than leaf developmental stage. E-mail mgarcia@zoo.uvm.edu; phone (802) 656-2824.

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R.E. Gaussoin, J.A. Murphy, and B.E. Branham

A method for measuring soil water potential in field soils was adapted for use in turfgrass soils. The system uses tensiometers installed flush with the soil surface and permits a measuring depth as shallow as 2.5 to 5.0 cm. Water potential within a tensiometer was measured with a portable pressure transducer. Linear relationships between water potential measured with mercury manometers or vacuum gauge-equipped tensiometers and the pressure transducer were obtained (r2 = 0.99 and 0.97, respectively). The system accurately measures soil water potential of turfgrass soils, while permitting routine cultural practices to be performed.

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W.J. Hill, J.R. Heckman, B.B. Clarke, and J.A. Murphy

Take-all patch, caused by Gaeumannomyces graminis (Sacc.) Arx. & D. Olivier var. avenae (E.M. Turner) Dennis (Gga), is a disease of creeping bentgrass (Agrostis stolonifera Huds.), which most often is associated with golf courses. Synthesis of ligneous and phenolic compounds by plants requires adequate Mn+2 and Cu+2 nutrition and may be a factor in disease resistance. An experiment was conducted on a creeping bentgrass fairway naturally infested with Gga to determine if foliar applications of Mn+2 (1.02 and 2.04 kg·ha–1 per application) and Cu+2 (0.68 kg·ha–1 per application) would reduce take-all severity. Prior to initiating treatments, soil pH was 6.4 and Mehlich-3 extractable Mn+2 and Cu+2 were 5 mg·kg–1 and 1.7 mg·kg–1, respectively. Manganese and copper sulfate treatments were initiated in July 1995 and foliarly applied every 4 weeks through 1997 with the exception of December, January, and February. Disease incidence was decreased from 20% on untreated turf to 5% with the high rate of MnSO4. For both years, turf treated with the high rate of Mn+2 had less disease than turf receiving the low rate of Mn+2. The application of CuSO4, however, did not influence disease development.

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M. Elena Garcia, C.R. Rom, J.B. Murphy, and K. Kugler

Light is important in the production of phenolic compounds because key enzymes in phenolic biosynthesis are induced by light, and because products of photosynthesis are used in the synthesis of phenolic compounds. It is known that light intensity decreases with increasing depth in apple tree canopies. The objective of this experiment was to determine how leaf position on a limb affects the total foliar phenolic content. Leaves from `Stark Spur Supreme Red Delicious' on C6 and M26 rootstocks were collected on 28 July and 2 Aug. 1996. Each tree was divided into two sides, east and west. Each side was divided into 3 areas; exterior, middle, and interior. From each area, leaves were collected and PAR, SLW, assimilation, total N, and total phenolics were measured. Leaf position on a limb was a significant parameter for all of the measured variables. PAR, SLW, assimilation, total N, and total phenolics were highest in leaves at the exterior of the canopy. The total foliar phenolic content of the exterior canopy leaves was 20% higher than that found in the interior canopy leaves. There was a significant correlation between SLW and total phenolic content/cm2(r 2 = 0.77; P < 0.05). Assimilation may be a limiting factor in phenolics production in apple trees because of the correlation between assimilation and total phenolic content/cm2 (r2=0.56, P < 0.05).

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M.E. Garcia, R.C. Rom, J.B. Murphy, and G.W. Felton

The foliar phenolic content of 21 apple cultivars was evaluated. Ten leaves were sampled randomly from three positions on current-season terminal shoots. Shoots were divided as tip, middle, and basal positions. The phenolic content was determined by spectrophotometric method (390 nm) using diphenylboric acid 2-aminoethyl ester as the reagent and caffeic acid as standard. Cultivars varied significantly in phenolic content. `Stark Ultra Red' had the highest amount, and `Liberty' had the lowest amount. Significant variations in the phenolic content due to leaf position were observed. Phenolic content was highest in leaves from the tip position, and it decreased toward the basal portion of the shoot. Factors affecting the phenolic content of apple cultivars will be investigated to determine apple × insect interactions.