Modification of matrix polysaccharides during avocado (Persea americana) fruit ripening: an assessment of the role of Cx-cellulase

The role of C_x-cellulase (EC 3.2.1.4) in fruit ripening and softening is unknown. In the present study, avocado ( Persea americana ) fruit, a rich source of C_x-cellulase, were examined to determine if the enzyme plays a role in ripening-related hemicellulose metabolism. Hemicelluloses (4 M alkali-soluble) from avocado fruit exhibited a very broad distribution of polymer sizes and an overall decrease in M_r during ripening. Polymers affected were primarily those of large M_r (relative molecular mass). The characteristic total hemicellulose M_r distribution and changes with ripening were also evident for xyloglucan (XG), a putative substrate for avocado C_x-cellulase. Hydrolytic activity toward hemicelluloses from preripe fruit was detected in crude buffer-soluble protein extracts derived from ripe avocado mesocarp tissue. XG was also degraded, and in a pattern similar to that observed during ripening. Purified C_x-cellulase also exhibited activity against specific components of isolated hemicelluloses; however, in contrast to the crude protein. C_x-cellulase alone was without influence on the M_r distribution of avocado XG. Protein depleted of C_x-cellulase was capable of moderate XG depolymerization. We conclude from the present studies that the enzyme C_x-cellulase is not involved in the ripening-related depolymerization of XG in avocado fruit.     

Baseline susceptibility of persea mite (Acari: Tetranychidae) to abamectin and milbemectin in avocado groves in Southern California

Persea mite, Oligonychus perseae Tuttle, Baker, and Abatiello, susceptibility to abamectin and milbemectin was evaluated in 2003 to determine baseline susceptibility levels in avocado groves in San Diego and Ventura Counties (California, USA) where more than 70% of the state’s avocado production is concentrated. Milbemectin has yet to be used in avocado production in California and abamectin has been available for use since 1999. Baseline susceptibility ratios (in relation to the most susceptible population) of five persea mite field strains to milbemectin varied 2.1- to 2.8-fold at the LC₅₀ and LC₉₀, respectively. The susceptibility of seven field strains to abamectin varied slightly more (2.1- to 3.5-fold) with one strain subjected to seven sprays over the past 4 years showing slight but significant separation of LC₅₀ and LC₉₀’s from the most susceptible strain, which is suggestive of the early stages of resistance to this product. Based on these data, baseline susceptibility levels are proposed that might be used to monitor for future persea mite resistance to these chemicals as their use in California avocado production continues.     

Evidence for the presence of a native,non-auxinic rooting promoter in avocado (Persea americana Mill.)

The presence of a rooting promoter in paratially purified extracts of avocado (Persea amricana Mill.) organs has been demonstrated using the mung bean rooting bioassay. Extraction with 80% methanol was followed by partition into diethyl ether, paper chromatography (PC) and 3 steps of thin layer chromatography (TLC). The number of roots induced by the rooting promoter in the absence of exogenous auxin was 5 to 7 times higher than that of the water control and 50% higher than by 4-(indol-3-yl) butyric acid (IBA) at its reported optimum concentration. Rooting of tomato, Coleus and young avocado cuttings was also enhanced by the rooting promoter. The rooting promoter was inhibitory in the wheat coleoptile section elongation bioassay for auxins and had slight inhibitory activity in the split pea stem curvature test.The biological properties of the avocado rooting promoter may be comparable to those of -(p-chlorophenoxy) isobutyric acid (PCIB) which acts as an anti-auxin in certain bioassays and, nevertheless, promotes the rooting of mung bean cuttings.     

Cloning and characterization of avocado fruit mRNAs and their expression during ripening and low-temperature storage

Differential sereening of a cDNA library made from RNA extracted from avocado (Persea americana Mill cv. Hass) fruit stored at low temperature (7°C) gave 23 cDNA clones grouped into 10 families, 6 of which showed increased expression during cold storage and normal ripening. Partial DNA sequencing was carried out for representative clones. Database searches found homologies with a polygalacturonase (PG), endochitinase, cysteine proteinase inhibitor and several stress-related proteins. No homologies were detected for clones from six families and their biological role remains to be elucidated. A full-length cDNA sequence for avocado PG was obtained and the predicted amino acid sequence compared with those from other PGs. mRNA encoding PG increased markedly during normal ripening, slightly later than mRNAs for cellulase and ethylene-forming enzyme (EFE). Low-temperature storage delayed ripening and retarded the appearance of mRNAs for enzymes known to be involved in cell wall metabolism and ethylene synthesis, such as cellulase, PG and EFE, and also other mRNAs of unknown function. The removal of ethylene from the atmosphere surrounding stored fruit delayed the appearance of the mRNAs encoding cellulase and PG more than the cold storage itself, although it hardly affected the expression of the EFE mRNA or the accumulation of mRNAs homologous to some other unidentified clones.AFRC Research Group in Plant Gene Regulation     

Acetoacyl-acyl carrier protein synthase from avocado: its purification,characterisation and clear resolution from acetyl CoA:ACP transacylase

-ketoacyl-ACP synthetase III (KAS III) has been purified from avocado using a six-step purification procedure. The enzyme, which is cerulenin-insensitive and thiolactomycin-sensitive, was assayed using a partial component reaction: acetyl CoA:ACP transacylase (ACAT) activity. KAS III activity is distinguished from ACAT activity on the basis that the former is highly stimulated by the addition of malonyl CoA in the presence of malonyl-CoA:ACP transacylase, and the latter is not. KAS III and ACAT activity have been separated from each other thus providing the first evidence that these two discrete activities exist in higher plants. Both of these enzymes have been implicated in the initial reactions of fatty acid synthesis.KAS III was purified 134-fold using a combination of PEG precipitation, Fast Q, ammonium sulphate precipitation, Phenyl Sepharose and ACP-affinity chromatography. The enzyme requires Triton X-100 for solubility and is highly salt sensitive. The subunit molecular mass of 37 kDa has been identified by SDS-PAGE. The results of gel filtration analysis are consistent with the native enzyme being homodimeric. The native molecular mass of KAS III is 69 kDa and that of ACAT 18.5 kDa. The enzyme has a pH optimum of 7.0–7.5, which is similar to the pH optimum of the ACAT reaction. The Km for acetyl CoA is 12.5 M and the Km for malonyl-ACP is 14M. Both KAS III and ACAT are sensitive to thiolactomycin inhibition. The results are discussed with respect to the potential role of acetyl CoA:ACP transacylase in plants.     

Isolation, culture and regeneration of avocado (Persea americana Mill.) protoplasts

Protoplasts were isolated from embryogenic suspension cultures derived from avocado (Persea americana Mill.) zygotic embryos and nucellus in an enzyme digestion solution consisting of 1% cellulase Onozuka RS, 1% Macerase R10, 0.2% Pectolyase Y-23, 0.7 M mannitol. 24.5 mM CaCl₂, 0.92 mM NaH₂PO₄ and 6.25 2-[N-morpholino]ethanesulfonic acid (1.5 ml) mixed with 0.7 M MS^–8P (2.5 ml). MS^-8P medium consisted of Murashige and Skoog salts without NH₄NO₃, 1 mg l^–1 thiamine HCl, 100 mg l^–1 myo-inositol, 3.1 g l^–1 glutamine and 8P organic addenda. Medium osmolarity was adjusted with 0.15 M sucrose and 0–0.55 M mannitol. Protoplast yields of 3.5×10⁶ protoplasts g^–1 were obtained. Growth and development of the protoplasts were significantly affected by osmolarity, nitrogen source, plating density and culture medium dilution. Under optimum conditions, proembryos developed directly from embryogenic protoplasts and subsequently into somatic embryos. Optimum conditions for somatic embryo development included the culture of protoplasts at a density of 0.8–1.6×10⁵ ml^–1 in 0.4 M MS^–8P for 2–3 weeks, followed by subculture in 0.15 M MS^–8P at a diluted density of 20–40× for 1 month in darkness to obtain somatic embryos. Mature somatic embryos were recovered on semisolid medium; however, a low frequency of plantlet recovery (≤1%) from protoplast-derived somatic embryos was observed. Received: 9 February 1998 / Revision received: 4 May 1998 / Accepted: 15 May 1998     

Differences among avocado cultivars in susceptibility to polyphagous shot hole borer (Euwallacea spec.)

Polyphagous shot hole borer (PSHB) (Euwallacea spec.; Coleoptera: Curculionidae: Scolytinae) has recently invaded southern California, USA. Along with its associated fungi, the beetle causes branch dieback and tree mortality in more than 200 host tree species, including avocado (Persea americana Mill., Lauraceae) and many important native, urban landscape, and nursery stock trees. As PSHB spreads into the avocado‐growing regions of southern California, there is a pressing need for more information for effective management of the insect/fungus complex. The objective of this study was to examine avocado cultivars to see whether there were differences in susceptibility to attack and gallery development from PSHB. We conducted choice and no‐choice trials with cut branches in the laboratory, artificially infested avocado saplings with PSHB, and also surveyed attack rates for two cultivars of field‐grown avocado subject to natural attack by PSHB. Laboratory and field trials showed similar patterns for preference among avocado cultivars indicated by beetle attack rates and gallery formation. Among the common commercial cultivars, Fuerte, Gwen, and Bacon most often had lower attack rates and lower rates of gallery formation. Zutano most frequently indicated high attack rates and much gallery formation, indicating that it may be the most susceptible to PHSB of the commercial cultivars.     

Twenty years of vegetation change in five long-unburned Florida plant communities

Abstract. Rates and directions of change over a 20-yr interval in five long-unburned (> 60 yr) plant communities were studied using multivariate analyses and compositional vectors. The study sites were located in fire and summer-drought adapted, xerophytic vegetation with many endemics on acidic, nutrient-poor, sandy soils in south-central peninsular Florida. Sizes of individual stems from 72 sets of nested permanent quadrats were measured in 1969, 1979, and 1989. Patterns of vegetation change differed by community. Flatwood and bayhead quadrats showed rapid increases in densities and basal areas of Persea borbonia (red bay). In the southern ridge sandhill community, evergreen clonal Quercus species (oaks) and Pinus clausa (sand pine) increased in dominance and grasses declined. Oaks (especially Q. geminata) also increased in importance in scrubby flatwoods. Sand pine scrub was relatively stable in composition, but experienced marked structural changes due to substantial sand pine mortality (18% during 1969–1979, 39% during 1979–1989). Compositional changes in the absence of fire were greatest whereas structural changes were least in southern ridge sandhill and scrubby flatwoods, both communities which normally receive frequent, recurrent fire. Compositional changes were lowest in sand pine scrub, which is normally infrequently burned. Classic successional patterns such as species replacement, decreases in density, and increases in basal area were generally lacking. Tree densities increased in two of four community types (southern ridge sandhill, scrubby flatwoods); while basal area declined in the flatwoods/bayhead and sand pine scrub sites. Directions of compositional vectors included divergent, opposing, and complex patterns, suggesting vegetation change in the absence of fire has a strong stochastic component.     

Un gisement a plantes des formations pliocènes des environs de Rasteau près Vaison-la-Romaine (Vaucluse): Stratigraphie,faune, étude des macrorestes végétaux,palynologie

A new Pliocene deposit with vegetal impressionswas discovered at Rasteau (near Vaison-la-Romaine, Vaucluse. The study of these impression and the sporopollinic analysis from many levels allows us to specify its stratigraphical position in the Pliocene of the Rhone valley. So, the above mentioned levels settled just before the Hautimagne deposit (Neogene zone MN 14 for the mammals). The macroflora and the microflora appear to be related with a temperate warm and wet climate.