Evaluation of Likelihood of Co-Occurrence of Erwinia amylovora with Mature Fruit of Winter Pear

ABSTRACT Phytosanitary concerns about fire blight prohibit export of U.S.-grown pears to some countries without this disease. To examine these concerns, we evaluated the potential for co-occurrence of Erwinia amylovora with mature, symptomless winter pear fruit by inoculation experiments and by survey of commercial orchards. Immature pear and apple fruit were inoculated in orchards with E. amylovora strain 153N as resuspended lyophilized cells or as ooze from diseased tissues. Regardless of inoculum source, population size of Ea153N on fruit declined by an order of magnitude every 3 to 4 days during the first 2 weeks after inoculation; at 56 days after inoculation, Ea153N was not detected, except on 1 of 450 fruit with 4 colony forming units (CFU). After inoculation of flowers, calyx-end survival of Ea153N on pear and apple fruit declined from high populations at petal fall to a few cells at harvest, with no detection of the pathogen after a 7-week cold storage. Migration of Ea153N into symptomless pear fruit from diseased branches was evaluated by enrichment assay and nested polymerase chain reaction of internal fruit core tissues; these assays failed to detect the pathogen in healthy fruit from diseased trees. At harvest, E. amylovora could not be detected on 5,599 of 5,600 fruit of d'Anjou pear sampled from commercial orchards in major production areas of the Pacific Northwest; one fruit yielded 32 CFU of the pathogen. Postharvest, mature pear fruit contaminated with Ea153N and subsequently wounded required a dose of >10,000 cells at the wound site to allow for persistence of the pathogen through a 7-week-cold storage. We conclude that epiphytic E. amylovora shows similar survival characteristics on both pear and apple fruit, this pathogen is not an endophyte within mature symptomless pear fruit, its presence is exceptionally rare on commercially produced fruit, and that epiphytic survival of E. amylovora through a postharvest chilling period is unlikely given the unrealistically high population size required for persistence.     

Comparison of lignin and cellulose solubilities in ionic liquids by COSMO-RS analysis and experimental validation

In this work, the solubilities of lignin and cellulose in ionic liquids (ILs) have been analyzed and compared by COSMO-RS simulation. The excess enthalpy of the IL + lignin/cellulose mixtures is evaluated as reference property to describe the affinity of lignin and cellulose for different ionic liquids, in terms of the intermolecular interactions contributing to their mixing properties. Thus, the anion plays the main role in the dissolution process of both lignin and cellulose, in which the hydrogen bonding forces are the major interactions. The most promising anions for the cellulose and lignin dissolution are acetate, formate and chloride, whereas different cations can be employed. Computational results indicated that a rational combination of the anions and cations allows designing ILs able to dissolve with different selectivity lignin and cellulose. Some of these solutions are tested in the laboratory to validate the model employed and the regenerated solids are analyzed by FTIR spectroscopy.     

Enhanced molecular identification of Botrytis spp. from New Zealand onions

Botrytis spp. associated with neck rot disease were isolated from New Zealand onions. The fungi were identified using molecular sequences of the ribosomal internal transcribed spacer (ITS) and intergenic spacer (IGS) regions, and the glyceraldehyde-3-phosphate dehydrogenase (G3PDH) gene. Analyses of the sequences showed that the majority of the isolates gathered in 2005–07 were B. aclada. A new high resolution melting analysis (HRMA) assay was developed that allowed fast and simple discrimination between B. aclada and other Botrytis spp. causing onion neck rot in New Zealand. To further verify these results, Botrytis isolates from New Zealand onions, stored in the International Collection of Microorganisms from Plants (ICMP), were also examined. Only a single isolate from the ICMP collection was B. aclada while two isolates were B. byssoidea, one B. squamosa and another closely related to Botryotinia porri. Identification of the remaining Botrytis isolates was more difficult; while IGS and ITS sequences indicated a close relationship to B. allii or B. byssoidea, a previously unreported intron insertion was observed at the 3′ end of the ribosomal small subunit gene in these isolates. No evidence of heterogeneity was observed in the G3PDH gene sequences, as might have been expected of the allodiploid B. allii, but the G3PDH sequence ruled out B. byssoidea as the identity of these isolates.     

植物生长调节剂对虎眼万年青'Chesapeake Snowflake'开花的影响

对从鳞茎生长的虎眼万年青'ChesapeakeSnowflake'植株,施加10,50μL2%的Fascina-tionTm,植株提早开花12d;而施加100,200μL2%的FascinationTM,植株提早开花9~10d;施加200mg/L的Pro-Gibb^?,植株提早开花12d,而施加25,50和100mg/L的Pro-Gibb^?,植株提早开花6~7d.从小鳞茎生长的虎眼万年青'ChesapeakeSnowflake'植株,施加10,50μL2%的FascinationTM,植株提早开花14d,而施加100,200μL2%的FascinationTM,植株提早开花10~11d;施加200mg/L的Pro-Gibb^?,植株提早开花14d,而施加25,50mg/L的Pro-Gibb^?,植株提早开花8~9d.两种植物生长调节剂对花茎的长度没有明显的影响。     

A contribution to the maintenance of grapevine diversity: The rescue of Tinta Castañal (Vitis vinifera L.), a variety on the edge of extinction

The arrival of powdery mildew, phylloxera, and then downy mildew from America in the late 19th/early 20th century, was one of the most important causes of the loss of grapevine diversity in Europe. Many varieties traditionally grown in small winemaking areas of Europe were substituted by direct producing hybrids or by a small number of varieties from other vine-growing areas. For geographic, economic, sociological, and cultural reasons, the north and northwest of Spain acts as a kind of refuge area where grapevine diversity is still high. At the Misión Biológica de Galicia (CSIC) research station a collection of living vines with a number of dispersed individuals belonging to old varieties was established in 1992. Many of these varieties had practically disappeared from vineyards and the majority existed only as centuries-old individuals.     

Estimation of daily milk yield of Nellore cows grazing tropical pastures

Beef cows’ milk yield is typically determined by measuring milk yield once daily and then doubling this value to estimate daily production. However, it is not known whether this is accurate. Thus, we aimed to determine the association between morning and afternoon milk yield in grazing Nellore cows. Eighty Nellore cows were used, with initial weight of 516.0?±?1.0 kg. The experiment was a completely randomized factorial scheme, with 20 replications and four treatments (i.e., +?or ??pre-partum supplementation in combination with +?or ??post-partum supplementation): PRMM—1 kg of supplement/cow/day for 90 days pre-partum; MMPS—1 kg of supplement/cow/day for 90 days post-partum; PRPS—1 kg of supplement/cow/day for 90 days pre-partum and 90 days post-partum; and MM—only mineral mix ad libitum during pre- and post-partum. Milk was sampled on days 45, 135, and 225 post-partum (early, middle, and late lactation, respectively). No effects were observed of pre- and post-partum supplementation on milk yield (P?>?0.05). The afternoon/morning proportion of 0.45 in the early third of lactation was higher than other stages, which had a proportion of 0.41 (P?<?0.05). Post-partum supplementation increased milk protein in the morning and afternoon milking (P?<?0.05). There was also no effect of pre- and post-partum supplementation on afternoon-morning proportion other milk components (P?>?0.05). We conclude that estimating daily milk production of grazing beef cattle by multiplying a once daily milking amount times two is not accurate. Under the conditions of this study, proportion of total daily production represented by the ratio of afternoon/morning milking was 0.45 in early lactation (first third) and 0.41 in mid- and late lactation.     

Bioavailability of Iron to Pseudomonas fluorescens Strain A506 on Flowers of Pear and Apple

ABSTRACT The addition of 0.1 mM FeCl(3) to a defined culture medium induces the bacterial epiphyte Pseudomonas fluorescens strain A506 (A506) to produce an antibiotic toxic to the fire blight pathogen, Erwinia amylovora. Consequently, because A506 is registered and applied as a commercial product to suppress E. amylovora before floral infection of pear and apple, the relative availability of iron to A506 on surfaces of pear and apple flowers is of potential significance. An 'iron biosensor' construct of A506 was developed by transformation with an iron-regulated promoter (pvd) fused to a promoterless ice nucleation reporter gene (inaZ). This construct, A506 (pvd-inaZ), established high populations on pear and apple flowers, ranging from 10(4) to 10(6) CFU/flower. In seven trials on pear and apple trees, A506 (pvd-inaZ) expressed high ice nucleation activity (INA) on flowers, indicating limited iron bioavailability or a low-iron environment unlikely to induce antibiotic production by A506. A506 (pvd-inaZ) also colonized flowers when mixed with chemicals containing iron: FeSO(4) or the iron chelates ferric ethylenediaminedi-(o-hydroxyphenyl-acetic) acid (FeEDDHA) and ferric diethylenetriamine pentaacetate (FeDTPA). These compounds represent an array of commercial iron formulations applied to foliage to avert iron chlorosis. Treatment of flowers with a mixture of A506 (pvd-inaZ) and 3 mM FeEDDHA or FeDTPA significantly decreased INA compared with flowers treated with A506 (pvd-inaZ) in water. Lower concentrations (0.3 mM) of FeEDDHA, however, did not consistently suppress INA. These results indicate that apple and pear flowers represent an iron-limited environment to A506 and that treatment with 3 mM FeEDDHA is needed to increase significantly the level of iron available to this bacterium.