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Allan B. Woolf, Christopher B. Watkins, Judith H. Bowen, Michael Lay-Yee, John H. Maindonald, and Ian B. Ferguson

`Hass' avocados (Persea americana Mill.) were heated in air at 25 to 46C for 0.5 to 24 hours and stored at 0, 2, or 6C. After storage, fruit were ripened at 20C and their quality was evaluated. In unheated fruit, external chilling injury occurred in fruit stored at 0 or 2C, hut not 6C. Chilling injury was also evident after storage at 2C in fruit heated at 34C, and to a lesser extent in fruit heated at 36C. A heat treatment (HT) of 38C for 3, 6, or 10 hours and 40C for 0.5 hour further reduced external chilling injury induced by storage at 2C. These HTs did not reduce internal fruit quality and resulted in more marketable fruit than unheated fruit stored at 6C. Low-temperature storage and HT slowed avocado ripening, resulting in longer shelf life after storage. In flesh tissue sampled directly after selected HTs, the levels of mRNA homologous to cDNA probes for two plant heat-shock protein (HSP) genes (HSP17 and HSP70) increased to a maximum at 40C and declined at higher temperatures. These increases in gene expression coincided with the extent to which HTs prevented chilling injury. Hot-air HTs confer significant protection against low-temperature damage to avocados.

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Diana L. Lange and Adel A. Kader

Preclimacteric avocado [Persea americana (Mill.) cv. Hass] fruit or fruit disks as well as fruit harvested in either June (midseason) or August (late season) and partially ripened were kept in air (21% O2 + 78% N2), 20% CO2 + 17% O2 (63% N2), or 40% CO2 + 13% O2 (47% N2) at either 10 or 20 °C. Ethylene production by preclimacteric fruit was completely inhibited during CO2 exposure, whereas there was only partial inhibition of ethylene production when partially ripened fruit were exposed. Compared to the fruit stored in air, O2 uptake of fruit stored in 20% CO2 was decreased by 20%, whereas the fruit stored in 40% CO2 showed 25% more O2 uptake than air-stored fruit. Fruit subjected to a storage regime of 40% CO2 at 10 °C followed by 2 d in air had the best visual quality. In general, climacteric fruit treated with 20% CO2 at 10 °C showed increased pyruvate dehydrogenase (PDH) activity and decreased cytochrome oxidase (CytOx) activity. Fruit stored in 40% CO2 had reduced CytOx activity compared to air-stored fruit, and PDH activity was variable depending on the harvest season of the fruit. Our results show that the effect of elevated CO2 on a given enzyme depends on concentration of CO2, duration of exposure, physiological state of the fruit, and type of tissue exposed.

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Diana L. Lange and Adel A. Kader

Partially ripened avocado [Persea americana (Mill.) cv. Hass] fruit harvested in either June or Aug. 1994 were kept at 10 °C in air (21% O2), 20% CO2 (17% O2, balance N2), or 40% CO2 (13% O2, balance N2) for 7 to 12 days and then were transferred to air at 10 °C for 2 to 3 days. Mitochondrial respiration was stimulated in response to elevated CO2 treatments at 10 °C. A shift to alternative pathway (Alt) respiration occurred on day 4 in experiments using avocados from both harvest dates, with a return to initial levels in only the 20% CO2-treated fruit (June-harvested fruit after return to air). Elevated CO2 at 20 °C decreased the in vitro O2 consumption of isolated mitochondria compared to mitochondria kept in air. The Alt pathway contributed less to the total O2 uptake of CO2-treated mitochondria compared to mitochondria kept in air. The respiratory control ratios of the CO2-treated fruit and mitochondria were higher and lower, respectively, than the air controls. Induction of 33 to 37 kD proteins (corresponding to the size of the alternative oxidase proteins) occurred in avocados after 4 days in 40% CO2. These results indicate that elevated CO2 has various effects depending on concentration, duration and temperature of exposure, and mitochondrial function of avocado fruit, such as increased and altered respiratory oxidation and up-regulation of alternative oxidase proteins.

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Diana L. Lange and Adel A. Kader

Changes in cytosolic and vacuolar pH, ATP, ADP, and the ATP : ADP ratio were measured in whole fruit or mesocarp disks of avocado [Persea americana (Mill.) cv. Hass] during brief exposures to elevated CO2. Intact climacteric fruit exposed to air (21% O2), 20% CO2 (17% O2, balance N2), or 40% CO2 (13% O2, balance N2) had cytosolic pH values of 7.0, 6.6, and 6.4, respectively, while mesocarp disks had cytosolic pH values of 6.9, 6.7, and 6.4, respectively. The ß-ATP levels of intact climacteric fruit exposed to 20% CO2 or 40% CO2 for 2 h were reduced by 25% or 43%, respectively, relative to air-exposed fruit. HPLC analysis of nucleotide phosphates from preclimacteric avocados revealed that ATP levels and the ATP : ADP ratio increased in 40% compared to the air-stored fruit. However, 1 day after transfer to air, the effects of elevated CO2 had dissipated. These modifications in cellular state could alter the activity of respiratory enzymes in fruit exposed to elevated CO2 atmospheres.

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Samuel Salazar-García, Elizabeth M. Lord, and Carol J. Lovatt

The developmental stage at which the shoot primary axis meristem (PAM) of the `Hass' avocado (Persea americana Mill.) is committed to flowering was determined. Three-year-old trees were subjected to low-temperature (LT) treatments at 10/7 °C day/night with a 10-h photoperiod for 1 to 4 weeks followed by 25/20 °C day/night at the same photoperiod. Before LT treatment, apical buds of mature vegetative shoots consisted of a convex PAM with two lateral secondary axis inflorescence meristems lacking apical bracts each associated with an inflorescence bract. Apical buds did not change anatomically during LT treatment. However, the 3- and 4-week LT treatments resulted in inflorescences at 17% and 83% of apical buds, respectively. Trees receiving 2 weeks or less LT, including controls maintained at 25/20 °C, produced only vegetative shoots. Apical buds of 2-year-old trees receiving 3 weeks at 10/7 °C plus 1 week at 20/15 °C produced 100% inflorescences. GA3(100 mg·L-1) applied to buds 2 or 4 weeks after initiation of this LT treatment did not reduce the number of inflorescences that developed. `Hass' avocado apical buds were fully committed to flowering after 4 weeks of LT, but were not distinguishable anatomically from those that were not committed to flowering.

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Zeinah A. El-Hamalawi and John A. Menge

At monthly intervals, plants and stem cuttings of avocado (Persea americana Miller) `Hass' grafted on `Barr Duke' rootstock and `Topa Topa' growing in a lathhouse were wounded and inoculated with the stem canker pathogen, Phytophthora citricola Sawada. The seasonal changes (measured monthly) in the extent of colonization of the avocado plants by P. citricola followed a periodic pattern, with two peaks of colonization during an annual growth cycle. Concentration of free amino acids and total soluble carbohydrates in the plant tissues followed a periodic pattern with two peaks similar to that of canker growth. Months were significantly different for canker size, free amino acids, and total soluble carbohydrates of the bark tissues. The extent of colonization was highest during May-June, after the first vegetative flush, and during November-December, after the second vegetative flush. Total free amino acids of the hark tissue was highly correlated with canker size (r = 0.89). Although the total soluble carbohydrate of the bark tissue was also elevated during the periods of canker development, it showed lower positive correlation (r = 0.45) with canker size. Plants were relatively resistant to colonization through March-April, during the first vegetative flush, and through August-September, during the second vegetative flush. Cankers formed on stem cuttings were generally larger than those of intact plants.

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Shimon Meir, Sonia Philosoph-Hadas, Giora Zauberman, Yoram Fuchs, Miriam Akerman, and Nehemia Aharoni

Fluorescent products (lipofuscin-like compounds) of lipid peroxidation, which accumulate with age, were extracted from `Fuerte' avocado (Persea americana Mill.) peels during ripening. Fractionation and analysis of these fluorescent compounds (FCs) was carried out by an improved method, based on separation of FCs from-chlorophyll by Sep-Pak silica cartridges. A sharp rise in FCs content was found 2 days after harvest in avocado fruits stored at 22C, and ethylene enhanced this rise 3-fold on the 4th day. The accumulation of FCs preceded by at leasts days the onset of climacteric ethylene and respiration and by 2 days the decrease in fruit firmness. Moreover, a 6-foId increase in the FCs concentration occurred during 1 to 2 weeks of storage at SC, but the avocado fruits did not show any other detectable signs of ripening. These results suggest that lipid peroxidation may be regarded as one of the earliest detectable processes occurring during fruit ripening. Thus, an increase of FCs in peel may be employed as a horticultural characteristic for estimating initiation of ripening in avocado fruit.

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Thomas L. Davenport, Petra Parnitzki, Sabine Fricke, and Melanie S. Hughes

Pollination was investigated in five avocado (Persea americana Mill.) cultivars during two seasons. In the first year, `Simmonds' and `Hardee' branches with inflorescences were covered with cheesecloth bags to prevent pollination by large flying insects during either or both the first (Stage I) and second (Stage II) floral openings. Adjacent, tagged branches were left open as controls. The proportion of pollinated Stage I flowers ranged from <1% in `Simmonds' to 9% in `Hardee.' Pollination rates in Stage II ranged from 15% in `Simmonds' to nearly 69% in `Hardee'. Pollination during Stage II was proportional to the number of white stigmas available during that stage. Stage II pollination rates for bagged flowers and open flowers were similar, even though large flying insects were barred from bagged flowers. In the second year, similar experiments on cultivars Simmonds, Tonnage, Tower 2, and Choquette provided results consistent with those obtained the previous year. Virtually no pollination occurred in bagged Stage I flowers in all cultivars tested, and ≈1% of the open Stage I flowers were pollinated. Pollination of bagged and open Stage II flowers was generally the same within cultivars. The percent pollination of Stage II flowers ranged from a mean of 4.3% to 35%, depending on cultivar. The results show that self-pollination during the Stage II floral opening is the primary means of pollination of commercial cultivars grown in Florida. Moreover, the presence of developing fruits on branches bagged during the flowering season demonstrated that fruit set can occur without pollination by large flying insects.

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David E. Crowley, Woody Smith, Ben Faber, and John A. Manthey

Methods for Zn fertilization of `Hass' avocado (Persea americana Mill.) trees were evaluated in a 2-year field experiment on a commercial orchard located on a calcareous soil (pH 7.8) in Ventura County, Calif. The fertilization methods included soil- or irrigation-applied ZnSO4; irrigation-applied Zn chelate (Zn-EDTA); trunk injection of Zn(NO3)2, and foliar applications of ZnSO4, ZnO, or Zn metalosate. Other experiments evaluated the influence of various surfactants on the Zn contents of leaves treated with foliar-applied materials and on the retention and translocation of radiolabeled 65ZnSO4 and 65Zn metalosate after application to the leaf surface. In the field experiment, tree responses to fertilization with soil-applied materials were affected significantly by their initial status, such that only trees having <50 μg·g–1 had significant increases in foliar Zn contents after fertilization. Among the three soil and irrigation treatments, ZnSO4 applied at 3.2 kg ZnSO4 per tree either as a quarterly irrigation or annually as a soil application was the most effective and increased leaf tissue Zn concentrations to 75 and 90 μg·g–1, respectively. Foliar-applied ZnSO4, ZnO, and Zn metalosate with Zn at 5.4, 0.8, and 0.9 g·liter–1, respectively, also resulted in increased leaf Zn concentrations. However, experiments with 65Zn applied to leaves of greenhouse seedlings showed that <1% of Zn applied as ZnSO4 or Zn metalosate was actually taken up by the leaf tissue and that there was little translocation of Zn into leaf parenchyma tissue adjacent to the application spots or into the leaves above or below the treated leaves. Given these problems with foliar Zn, fertilization using soil- or irrigation-applied ZnSO4 may provide the most reliable method for correction of Zn deficiency in avocado on calcareous soils.

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Robert L. Heath, Michael V. Mickelbart, Mary Lu Arpaia, Claudia Fassio, and Ruby Miller

Vapor pressure deficit (VPD) is the driving force for plant water loss. However, air relative humidity (RH) can be used as a surrogate for VPD. While plants can adapt to environments with varying RH, little is known about how they respond to sudden shifts in RH. Areas of Southern California can experience drastic shifts in RH, from 60% or greater to less than 20% in just a few hours. The effect of these shifts on avocado (Persea americana Mill.) tree productivity is a major concern to growers. We studied the effect of shifts in RH on `Hass' avocado leaf stomatal conductance (g s) and branch sap flow in trees grafted on Duke 7 clonal rootstock. Under many conditions, the avocado assimilation rate is governed by g s. When g s is high in morning (>150 mmol·m-2·s-1), the water loss generally leads to some stomatal closure in the afternoon (50% or more). Conversely, low morning g s results in a higher g s rate in the afternoon (10% to 20% stomatal closure). This relationship between morning and afternoon g s is intensified by a shift from high to low RH in the afternoon. Therefore, in a drier atmosphere in the afternoon, the afternoon depression in g s is greater, leading to an impaired assimilation capacity. We hypothesize that the afternoon decrease in g s is due to low root/shoot hydraulic conductivity since soil water is readily available. While it is possible that low hydraulic conductivity on g s is exacerbated at the graft union, sap flow of grafted trees in greenhouse studies was nearly equal to trees on their own roots (ungrafted); in fact, often the depression in the afternoon was less on grafted trees. This suggests that while avocado is not suited to areas with low RH, water flow through the roots could be an additional criterion in selecting improved rootstocks.