Molecular typing of ‘<Emphasis Type="Italic">Candidatus</Emphasis> Phytoplasma mali’ and epidemic history tracing by a combined T-RFLP/VNTR analysis approach

Apple proliferation caused by ‘Candidatus Phytoplasma mali’ is a disease of apple trees gaining increasing importance in Europe. The present study describes a high-throughput method for simultaneous typing of ‘Ca. P. mali’ at two genetic loci. This novel approach combines terminal restriction fragment length polymorphism (T-RFLP) analysis of a putative rhodanese-like protein gene and the analysis of the variable number of tandem repeats (VNTR) of the ribosomal protein L22 gene. The typing approach was applied to analyse a collection of DNA isolates from 310 apple trees tested positive for ‘Ca. P. mali’. Samples were taken between 2002 and 2010 in South Tyrol (Northern Italy). In addition, 15 samples of Cacopsylla melanoneura and 19 of C. picta were typed. Seven combined genetic profiles were found in the samples of infected apple trees: AT-2/rpX-A (81.0%), AT-1/rpX-D (8.4%), AT-1/rpX-E (4.2%), AT-1/rpX-A (3.2%), AT-1/rpX-B (1.6%), AT-1/rpX-C (1.0%) and AP/rpX-A (0.3%), and one mixed infection AP + AT-1/rpX-A + rpX-D (0.3%). Subtype rpX-E was discovered for the first time. In C. melanoneura samples the most frequent subtype was AT-1/rpX-E, followed by AT-1/rpX-D and AT-1/rpX-C. All C. picta samples displayed subtype AT-2/rpX-A. Analysis of the temporal distribution of subtype frequencies in apple trees revealed that exclusively subtype AT-1 in combination with four rpX subtypes was present in South Tyrol in the period from 2002 to 2004. From 2006 onwards subtype AT-2/rpX-A became dominant with an average frequency of 90%. The data obtained suggest that there may be a co-adaptation of particular ‘Ca. P. mali’ subtypes with different insect vector species.     

Seasonal colonisation of apple trees by ‘Candidatus Phytoplasma mali’ revealed by a new quantitative TaqMan real-time PCR approach

Apple proliferation (AP), caused by ‘Candidatus Phytoplasma mali’, is an economically important disease affecting many apple-growing areas in Europe. A new TaqMan real-time PCR assay was established for absolute quantification of ‘Ca. P. mali’ by using a single-copy gene of the host plant as a reference, which is amplified with the pathogen DNA in a single-tube reaction. Normalised estimates of phytoplasma concentration are ultimately expressed as the number of phytoplasma cells per host plant cell. The assay was used to monitor the ‘Ca. P. mali’ titre over the course of two growing seasons in roots and branches of symptomatic and asymptomatic but AP-positive apple trees. All 252 root samples from symptomatic and asymptomatic trees tested positive, with an average number of 59.8 ± 5.68 (standard error) and 55.1 ± 9.83 ‘Ca. P. mali’ per host cell, respectively. From the 378 shoot samples analysed, 81% of the symptomatic and only 20% of the asymptomatic samples were AP-positive with an average number of 9.4 ± 1.04 and 0.7 ± 0.13 ‘Ca. P. mali’ per host cell, respectively. This strengthens evidence that not the pathogen occurrence alone but the presence of a certain quantity of ‘Ca. P. mali’ in the aerial tree sections is involved in symptom expression. In addition, pronounced seasonality of the phytoplasma concentration was found, not only in branches, but also for the first time in roots of symptomatic and asymptomatic apple trees. Highest phytoplasma levels in roots were detected from December to May.     

Waiting reveals waning weight: Monitoring over 54 years shows a decline in body condition of a long-lived reptile (tuatara, Sphenodon punctatus)

Knowledge gained from monitoring has been the basis for many critical decisions in threatened and endangered species and ecosystem management. Long-term monitoring has been recognized as a necessity for elucidating population trends and community interactions, particularly for long-lived species or ecosystems with slow rates of change. We examine seasonal and annual cycles of morphological changes in a threatened, long-lived, insular reptile, the tuatara (Sphenodon punctatus), on Stephens Island, New Zealand. We used body condition indices as a surrogate measure of fitness, and examined seasonal fluctuations, using data from an intensive mark-recapture study, and long-term trends using a dataset that spans 54 years. In spite of seasonal and annual fluctuations, body condition of tuatara has declined significantly between 1949 and 2003; the decline was only evident after >22 years of monitoring. We hypothesize that increasing numbers of tuatara have resulted in a density-dependent population response driven by past habitat modification on Stephens Island. We emphasize the need for long-term monitoring and suggest that potentially costly management decisions, particularly for long-lived species or ecosystems with slow rates of change, should not be based solely on short-term monitoring.     

Phenotypic recurrent selection in ideotype breeding of pinto beans

Summary Plant growth habit (type) plays a major determining role in the acceptance of a dry bean cultivar by commercial growers. Under the threat of wet fall conditions in the midwestern U.S., growers show a preference for upright plant types-I and II over the more prostrate type-III growth habit of commercial pinto cultivars. The prostrate habit creates considerably more risk of harvest losses and potential of white mold (Sclerotinia sclerotiorum) development during a wet growing season.The breeding strategy being utilized to develop better adapted pinto bean cultivars for the humid midwest is based on the concept of ideotype breeding which has been successfully applied to small seeded navy and black beans. A system of phenotypic recurrent selection using S₁ selection is being followed in order to recombine desirable traits from two diverse germplasm pools. One germplasm pool possesses the desirable architectural plant form and disease resistance traits of the small-seeded types while the medium-seeded germplasm source possesses the desirable seed size, shape and color characteristics of the pinto class. Since a repulsion phase linkage appears to exist between type-II architecture (architype) and medium seed size (40 g/100 seeds), recurrent selection is shown to be the most effective procedure for breaking up undesirable linkage groups and for increasing the frequency of desirable genes, thus enhancing the chance for the desirable genetic recombinations to occur.Journal Article No. 11667 of the Michigan Agricultural Experiment Station.     

Sorption of cupric,dichromate and arsenate ions in some New Zealand soilds

Concentrated solutions of copper (Cu²⁺), dichromate (Cr₂O^2? ₇) and aresenate (AsO₄ ^3?) ions (CCA solutions) are used extensively in the New Zealand timber preservation industry. These ions are therefore, potential soil pollutants at timber treatment sites. Sorption of these three ions was examined by the surface and sub-surface horizons of two free-draining New Zealand soils over a range of soil solution pH values. Copper sorption by both soils increased substantially with increasing pH and was greater in the surface compared with the sub-surface horizons. Less dichromate was sorbed than the other two ions and wa similar in both surface and sub-surface horizons for each soil. Dichromate sorption increased with decreasing pH. Arsenate sorption from solutions containing all three ions was not greatly different to influenced by changes in soil solution pH. Arsenate sorption was generally greater in the sub-surface horizons of both soils. Sorption from solutions containing all three ions was not greatly different to sorption from solutions containing the single metal ions. Sorption behaviour for each ion is related to its chemistry and the soil chemical properties of each horizon. Results suggest that in the event of soil contamination by CCA solution, the immediate leaching potential of the initial ions species present would increase in the following order: Cu²⁺ < HAsI ₄ ^? ? Cr₂O ₇ ^2? .     

Influence of strawberry yogurt composition on aroma release

The primary objective of this study was to determine how yogurt ingredients affect aroma release in the mouth during eating. A model strawberry flavor consisting of ethyl butanoate, ethyl 3-methylbutanoate, (Z)-hex-3-enol, 2-methylbutanoic acid, 5-hexylhydro-2(3H)-furanone, and 3-methyl-3-phenylglycidic acid ethyl ester was added to unflavored, unsweetened yogurt that had different added sweeteners and hydrocolloids. In all, 12 yogurt formulations were examined to determine the effects of gelatin, modified food starch, pectin, sucrose, high-fructose corn syrup, and aspartame on aroma release. Aroma release was monitored by breath-by-breath analysis (proton-transfer reaction-mass spectrometry) during eating of the test yogurts. Results showed aroma release of the ethyl butanoate, (Z)-hex-3-enol, and ethyl 3-methylbutanoate to be suppressed by sweeteners, with 55 DE high-fructose corn syrup having the greatest effect. Addition of thickening agents had no significant effect on the aroma release profiles of the compounds under study.     

Effect of roasting on the antioxidant activity of coffee brews

Colombian Arabica coffee beans were roasted to give light, medium, and dark samples. Their aqueous extracts were analyzed by gel filtration chromatography, UV-visible spectrophotometry, capillary electrophoresis, and the ABTS(*)(+) assay. A progressive decrease in antioxidant activity (associated mainly with chlorogenic acids in the green beans) with degree of roasting was observed with the simultaneous generation of high (HMM) and low molecular mass (LMM) compounds possessing antioxidant activity. Maximum antioxidant activity was observed for the medium-roasted coffee; the dark coffee had a lower antioxidant activity despite the increase in color. Analysis of the gel filtration chromatography fractions showed that the LMM fraction made a greater contribution to total antioxidant activity than the HMM components.