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  • Author or Editor: Mark Rieger x
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The blueberry industry experiences significant losses every year due to environmental factors with a dehydrative component, such as drought and freezing stress. In a recent survey of blueberry research and extension scientists in the United States, lack of cold hardiness and susceptibility to spring frosts were identified as the most important genetic limitations of current cultivars. For these reasons, cloning and characterization of expression of dehydration-responsive genes (dehydrins) have been ongoing in our laboratory. To date, one full-length (2.0 kb bbdhn1 gene) and four partial-length dehydrin cDNAs have been cloned and sequenced. Very high homology at the DNA and protein levels were found among the blueberry dehydrin clones, particularly at the 3' ends. From DNA blots, it appears that blueberry dehydrins are encoded by about three genes with high homology to the full-length 2.0 kb bbdhn1 cDNA clone and a few other less related genes. The 2.0 kb bbdhn1 gene was mapped in a blueberry population segregating for cold hardiness and chilling requirement. Expression studies indicated that dehydrins are induced by cold and drought stress. In general, dehydrins were induced in all organs examined in response to cold stress, including floral buds, leaves, stems, and roots. Under drought conditions, dehydrins were induced primarily in stems and their levels declined in roots. Some of the induced dehydrins were the same for drought and cold stress, whereas others were unique to a given stress. Levels of dehydrin accumulation correlated positively with cold hardiness; however, levels of dehydrins did not correspond precisely to the degree of drought avoidance.

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Abstract

Two-year-old ‘Hamlin’ orange trees [Citrus sinensis (L.) Osb.],on Poncirus trifoliata (L.) Raf. rootstock were used to study the effects of various microsprinkler irrigation rates and spray patterns on trunk temperatures and microclimate during freeze conditions. Irrigation treatments consisted of a factorial combination of two spray patterns (90° and 360°) × three application rates (38, 57, and 87 liters hr−1), plus an unirrigated control. All trunks were wrapped with fiberglass insulation. Trunk temperatures were 1° to 5°C higher for irrigated than unirrigated trees, regardless of spray pattern or application rate and were 1° to 2°C higher with the 90° than 360° spray pattern. The 57 and 87 liter·hr−1 rates produced the highest trunk temperatures, although 38 liter·hr−1 maintained trunk temperatures above −2.5°C with air temperatures as low as − 12°C. Outgoing radiation was higher for irrigated than unirrigated treatments during advective freeze conditions only, but air temperature and vapor pressure were unaffected by microsprinker irrigation.

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In a series of experiments, gibberellic acid (GA3) was applied to rabbiteye blueberries (Vaccinium ashei Reade) under field and greenhouse conditions to determine if fruit set could be improved following physical or freeze injury to flowers. In field experiments, physically damaged flowers (i.e., corollas and styles removed, styles only removed, or ovaries lanced) of `Climax' and `Tifblue' treated with GA3 (4% ProGib at 250 mg·liter–1) set substantially more fruit than nontreated, damaged flowers. Under green-house conditions, GA3 applied postfreeze to `Tifblue' and `Brightwell' resulted in increased fruit set compared to unsprayed control plants of the same cultivars. Freeze-damaged plants had substantially reduced fruit set overall but to a much lesser extent for GA3-treated plants than for those not treated with GA3. Individual fruit weight was reduced by GA3 applications, as was berry seediness. Results from these greenhouse and field trials suggest that GA3 can be used to salvage a blueberry crop following a moderate freeze during bloom.

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Container-grown Viburnum plicatum var. tomentosum `Mariesii' were planted in tilled beds and tilled beds amended with aged pine bark. After transplanting, plants were fertilized at three different rates: no fertilizer, 18.4 g of N m-2, and 36.8 g of N m-2. A 31 day drought was begun 73 days after planting. Fertilization of tilled plots induced ammonium toxicity, which caused a linear reduction in leaf area, shoot dry weight, and root dry weight. Fertilization of amended plots had no effect on shoot growth but reduced mot growth by 54%; thus, amendments ameliorated ammonium toxicity. Between 10 and 28 days after beginning the drought, plants in unfertilized-amended plots maintained higher relative leaf water contents (RLWC) and relative leaf expansion rates (RLER) than plants in unfertilized-tilled plots. Amendment induced nitrogen deficiencies contributed to the increased drought tolerance of plants from unfertilized-amended plots. Since fertilized plants developed symptoms of ammonium toxicity, we were unable to determine if increasing fertility would counteract the drought tolerance conferred by pine bark soil amendments.

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Isolite is a ceramic-like, porous soil amendment purported to sustain plant growth under reduced irrigation and increase plant survival during drought. The purpose of this greenhouse experiment was to determine the effect of an Isotite-amended soilless container medium on: (1) growth under reduced irrigation frequency and (2) water stress during drought of Impatiens × hybrids `Accent Red'. On 2 June 1993, seedlings were transplanted into 13.2 liter black plastic pots containing a 4:1 composted pine bark:coarse sand (vol.) medium amended with Isolite CG-1 granules at rates of 0%, 10%, 15%, and 20% (vol.). Study I. Seedlings were. irrigated with 500 ml tap water every two days for two weeks followed by a 4 week schedule of 500 ml tap water every 1, 2, 3, 4, or 5 days. In general, growth parameters were explained by irrigation treatment effects and did not differ with Isolite rate. Growth indices ranged from +54% to + 143%, while final visual quality grades ranged from 2.4 to 5.0 (5-point scale), shoot dry weight from 8.7 to 30.7 g, root dry weight from 2.0 to 7.9 g, and leaf area from 0.14 to 0.48 m2. Study II. Seedlings were irrigated with 500 ml tap water every 2 days for rive weeks followed by a two week drought. Plant water status parameters were similar at all rates of Isolite. Leaf expansion rates ranged from + 89% to +98%, white a final mid-day xylem pressure potential of -0.4 MPa and a final visual quality grade of 2.0 were uniform across all treatments. Under these conditions, Isolite did not limit water stress of container-grown Impatiens `Accent Red'.

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Sorbitol is the major photosynthetic product in peach. In sink tissues, sorbitol is converted to fructose via the NAD-dependent enzyme sorbitol dehydrogenase (SDH). A new assay is described that allows rapid, simple quantitation of SDH activity in growing shoot tips, root tips, and fruits. The activity was measured on the crude extract desalted with a Saphadex G-25 column to eliminate small molecules such as sugars and nucleotides. Optimum buffer type and pH for the enzyme as well as degradation by proteolytic enzymes and stability over time were determined in the present study. Inhibition by dithiothreitol (DTT) was detected at an inhibitor concentration as low as 2 mM, proving the similarity with mammalian SDH. Storage of samples at 4°C overnight resulted in significant loss of enzyme activity. Using this assay, we also correlated SDH activity with sink strength in peach.

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Sixteen cultivars of citrus (Citrus spp.) and close citrus relatives were planted in Savannah, Georgia to evaluate their potential as fruiting landscape trees in an area that routinely experiences minimum temperatures of 15 to 20 °F (-9.4 to -6.7 °C) during winter. Three to six trees of each cultivar were planted in 1998, and stem dieback and defoliation data were collected in 1999, 2001, and 2002. During the 4 years of the study, air temperatures fell below 32 °F (0.0 °C) 27 to 62 times per season, with absolute minima ranging from 13 to 18 °F (-10.6 to -7.8 °C), depending on year. In general, kumquats (Fortunella spp.), represented by `Meiwa', `Nagami', and `Longevity', were completely killed (or nearly so) in their first year in the field after air temperature minima of 13.5 °F (-10.28 °C). Others experiencing 100% dieback were `Meyer' lemon (Citrus limon × C. reticulata) and `Eustis' limequat (C. aurantifolia × Fortunella japonica), which were tested twice during the study. Kumquat hybrids, including procimequat [(C. aurantifolia × F. japonica) × F. hindsii), `Sinton' citrangequat [(C. sinensis × Poncirus trifoliata) × unknown kumquat], `Mr John's Longevity' citrangequat [(C. sinensis × P. trifoliata) × F. obovat], razzlequat (Eremocitrus glauca × unknown kumquat), and `Nippon' orangequat (C. unshiu × F. crassifolia) survived freezing, but all experienced at least some defoliation and stem dieback. `Owari' satsuma (C. unshiu), `Changsha' mandarin (C. reticulata), nansho daidai (C. taiwanica) and ichang papeda (C. ichangensis) experienced only minor stem dieback but substantial defoliation in most years, except that ichang papeda was substantially damaged in the last year of the study. Seven cultivars produced fruit at least once during their first 4 years: nansho daidai, ichang papeda, `Nippon' orangequat, `Mr John's Longevity' citrangequat, `Owari' satsuma, `Changsha' mandarin, and procimequat. Based on cold hardiness, fruiting, and growth characteristics, `Owari' satsuma, `Changsha' mandarin, `Mr John's Longevity' citrangequat, and `Nippon' orangequat provided the hardiest, most precocious and desirable fruiting landscape trees in this study.

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Illiciums, or star-anises, have increased in popularity in the nursery and landscape industries. However, confusion exists as to which taxa are tolerant of high light intensities during production and subsequent establishment in the landscape. We investigated the effect of two light intensity treatments, 45% and 100% full sunlight, on gas-exchange parameters of five Illicium taxa: Illicium anisatum L., I. floridanum Ellis. `Pebblebrook', I. henryi Diels., I. lanceolatum A.C. Sm., and I. parviflorum Michx. Ex. Vent. `Forest Green'. Light-response curves were determined for individual leaves, and mean response parameters calculated. Chlorophyll and total carotenoids were analyzed after extraction in acetone, with total chlorophyll also estimated with a SPAD chlorophyll meter. In general, highest rates of CO2 assimilation (Amax) and lowest rates of dark respiration (Rd) were found in the 45% light treatment for all taxa. Both Illicium anisatum and I. floridanum `Pebblebrook' had substantial reductions in Amax in 100% light, 94% and 81% respectively, compared to plants grown in the 45% light treatment. Illicium henryi failed to survive the 100% light treatment. Illicium lanceolatum and I. parviflorum `Forest Green' were least affected by the 100% light treatment. Severe photooxidative bleaching was noted and confirmed by SPAD and pigment data, although SPAD readings were a poor predictor of total chlorophyll. For taxa of Illicium in our study, photosynthetic gas-exchange parameters and foliage pigment characteristics were improved in the low light treatment, suggesting optimal growth occurs in shaded conditions.

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Terminal portions of `Flordaguard' peach roots [Prunus persica (L.) Batsch] were divided into six segments and the activities of NAD+-dependent sorbitol dehydrogenase (SDH), sorbitol oxidase (SOX), sucrose synthase (SS), soluble acid invertase (AI), and soluble neutral invertase (NI) were measured in each segment 10, 15, and 20 days after seed germination. The same type of experiment was conducted with terminal portions of `Flordaguard' and `Nemaguard' peach shoots except that one of the six segments consisted of the leaflets surrounding the apex. Independent of the age of individual roots, activities of SDH and AI were consistently highest in the meristematic portion and decreased with tissue maturation. In shoots, AI was the most active enzyme in the elongating portion subtending the apex, whereas SDH was primarily associated with meristematic tissues. A positive correlation between SDH and AI activities was found in various developmental zones of roots (r = 0.96) and shoots (r = 0.90). Sorbitol and sucrose contents were low in roots regardless of distance from tip, while sucrose showed a decreasing trend with distance and sorbitol, fructose, and glucose increased with distance from the meristem in shoots. Activity of SDH in internodes, but not apices, correlated with shoot elongation rate of both cultivars, whereas activities of other enzymes did not correlate with shoot elongation rate. We conclude that AI and SDH are the predominant enzymes of carbohydrate catabolism and the best indicators of sink growth and development in vegetative sinks of peach.

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Sorbitol is the major photosynthetic product in peach [Prunus persica (L.) Batsch.]. In sink tissues, sorbitol is converted to fructose via NAD+-dependent SDH. A new procedure is described that allows rapid, simple quantification of SDH activity in growing tissues. The procedure uses only 0.01 to 5 g of fresh tissue per sample, such that a single shoot tip, a single root tip, or ≈5 g of fruit flesh can be assayed for SDH activity. Storage of samples at 4 or -20 °C overnight resulted in significant loss of enzyme activity. Thus, freshly harvested tissues were ground with sand in buffer at 2 °C in a mortar and pestle, and the homogenate was centrifuged at 3000 g n to remove particulate matter and sand. The supernatant was desalted on a Sephadex G-25 column, and the eluent was assayed for SDH activity immediately. Activity was determined by measuring the production of NADH per minute in the assay mixture using a spectrophotometer (340 nm). Tris buffer at pH 9.0 was the best for extraction of peach SDH. Activity of SDH was strongly inhibited by dithiothreitol (DTT) in the extraction mixture and by DTT, L-cysteine, or SDI-158 in the assay mixture, similar to results reported for SDH from mammalian tissues. Peach SDH has a Km of 37.7 mm for sorbitol and a pH optimum of 9.5, similar to those reported for apple (Malus × domestica Borkh.) SDH. Unlike older protocols for SDH activity in plant tissues, the new procedure features reduced sample size (1/10 to 1/100 of that which was previously used), smaller volumes of buffer, fewer buffer ingredients, greatly reduced time for sample preparation, yet comparable or higher values of SDH specific activity. Following the same procedure, SDH activity was also measured in Prunus fremontii Wats., Prunus ilicifolia (Nutt.) Walp., and Marianna 2624 plum (P. cerasifera Ehrh. × P. munsoniana Wight & Hedr.).

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