Search

Several of the campus heritage plans funded by the Getty Foundation served as laboratories for applying the relatively new field of cultural landscape preservation to campus planning. With a strong landscape component, the heritage plans of The University of Kansas, Cranbrook Academy, the University of California, Berkeley, and elsewhere remind people that landscape preservation is inherently different from architectural preservation--and that the young field remains far richer in edifying puzzles than proven answers. The Getty Foundation-funded plans also remind everyone that planners should always challenge the assumed wisdom about what is "historic" and how to steward it. In several of the completed campus heritage plans, one can find site-specific decisions regarding some of these landscape preservation dilemmas. Yet one should note that these strategies are "choices made" and not universal solutions. The author argues that there remains no one set of best practices in campus landscape preservation--and this lack of resolution may be why its puzzles can enrich heritage preservation discourse overall. (Contains 11 figures and 1 note.)

Macrophomina crown and root rot has become a significant soil-borne disease issue in California. For many locations in the state, the disease is associated with fields that are no longer pre-plant, flat field fumigated with methyl bromide + chloropicrin. Inoculation experiments indicated that some differences in strawberry cultivar susceptibility toMacrophomina phaseolinawere seen a short time after the inoculation, but as disease progressed such differences did not persist. Preliminary characterization studies ofMacrophomina phaseolinaisolates from strawberry indicated that such isolates may have a host preference for strawberry.Macrophomina phaseolinaisolates from watermelon, thyme, and apple failed to cause disease in strawberry. Five cover crop species, which can be rotated with strawberry, did not develop disease when inoculated with strawberry isolates. In preliminary analysis using simple sequence repeat markers, isolates obtained from strawberry formed a separate group compared to isolates recovered from other knownMacrophomina phaseolinahosts. [ABSTRACT FROM PUBLISHER]

The activity of commercial soil fumigants on some key soilborne pathogens was assessed in sandy loam soil under controlled conditions. Seven soil fumigants that are registered in California or are being or have been considered for registration were used in this study: dimethyl disulfide (DMDS) mixed with chloropicrin (Pic) (79% DMDS and 21% Pic), Tri-Con (50% methyl bromide and 50% Pic), Midas Gold (33% methyl iodide [MI] and 67% Pic), Midas Bronze (50% MI and 50% Pic), Midas (MI, active ingredient [a.i.] 97.8%), Pic (a.i. 99% trichloronitromethane) and Pic-Clor 60 (57% Pic and 37% 1,3-dichloropropene [1-3,D]). Dose-response models were calculated for pathogen mortality after 24 hours of exposure to fumigants. Overall, the tested fumigants achieved good efficacy with dosages below the maximum label rate against the tested pathogens. In this study, Pythium ultimum and citrus nematode were sensitive to all the fumigants and Verticillium dahliae was resistant. For most fumigants, California regulations restrict application rates to less than the maximum (federal) label rate, meaning that it is possible that the fumigants may not control major plant pathogens. This research provides information on the effectiveness of these alternatives at these lower application rates. The results from this study will help growers optimize application rates for registered fumigants (such as Pic and 1,3-D) and will help accelerate the adoption of new fumigants (such as DMDS) if they are registered in California. [ABSTRACT FROM AUTHOR]

A 5-year project to facilitate the adoption of strawberry production systems that do not use methyl bromide initially focused on fumigant alternatives and resulted in increased use of barrier films that reduce fumigant emissions. The focus shifted in year 3 to evaluating and demonstrating nonfumigant alternatives: soilless production, biofumigation, anaerobic soil disinfestation (ASD) and disinfestation with steam. In the 2010-2011 strawberry production season, fruit yields on substrates were comparable to fruit yields using conventional methods. Anaerobic soil disinfestation and steam disinfestation also resulted in fruit yields that were comparable to those produced using conventionally fumigated soils. Additional work is in progress to evaluate their efficacy in larger-scale production systems in different strawberry production districts in California. [ABSTRACT FROM AUTHOR]

The mitochondrial genome of an isolate of Phytophthora ramorum from Europe (EU) was sequenced and compared to the previously published genome sequence of an isolate from California (NA). The EU mitochondrial genome had the identical gene order and encoded for the same suite of genes as the NA mitochondrial genome, but had 13 single nucleotide polymorphisms (SNPs) and at 39,494 bp was 180 bp longer. This length difference was due to an increase in the size of the spacer region between the nad5 and nad6 genes caused by a chimeric region containing duplication of the spacer sequence and additional sequences from the flanking genes. Recombination between the 1,150 bp-inverted repeats (IR) generated orientational isomers where the gene order was reversed between the IR. A total of seven primer pairs were developed for amplification of regions where the SNPs were located and two other regions where additional SNPs were encountered when a larger number of isolates were examined. Sequence data for a total of 5,743 bp for 40 isolates collected from a range of geographic areas was compared and 28 loci were found to be polymorphic. The combination of these polymorphisms revealed a total of 4 mitochondrial haplotypes; the traditional EU (haplotype I), the traditional NA (haplotype IIa), the third nuclear lineage of the pathogen recovered from a nursery in Washington State (haplotype III) and a new haplotype representing a subgroup of NA isolates from an Oregon forest (haplotype IIb). Phylogenetic analysis using the sequences generated from the haplotype analysis supported a high affinity for haplotypes IIa and IIb, both of which were distinct from haplotype I, with haplotype I basal to these and haplotype III representing the ancestral state. [ABSTRACT FROM AUTHOR]

The recent phase-out of the soil fumigant methyl bromide (MB) due to its impact on stratospheric ozone presents a huge challenge to strawberry nursery producers. We evaluated the effectiveness of alternative fumigants on soil pests and plant productivity, as well as production costs in California strawberry nurseries. Our trials followed nursery stock through low- and high-elevation phases of runner-plant propagation and a complete cycle of fruit production in coastal fields. Plant yields from the nurseries and fruit yields from Oxnard and Watsonville indicated that nursery plots treated with iodomethane plus chloropicrin, with 1,3-dichloropropene followed by dazomet, and with chloropicrin followed by dazomet produced runner-plant yields that were similar to methyl bromide plus chloropicrin. However, our economic analysis suggests that nursery profitability may nonetheless suffer from the loss of methyl bromide. [ABSTRACT FROM AUTHOR]

Sudden oak death is a disease currently devastating forest ecosystems in several coastal areas of California. The pathogen causing this is Phytophthora ramorum, although species such as P. nemorosa and P. pseudosyringae often are recovered from symptomatic plants as well. A molecular marker system was developed based on mitochondrial sequences of the coxI and II genes for detection of Phytophthora spp. in general, and P. ramorum, P. nemorosa, and P. pseudosyringae in particular. The first- round multiplex amplification contained two primer pairs, one for amplification of plant sequences to serve as an internal control to ensure that extracted DNA was of sufficient quality to allow for polymerase chain reaction (PCR) amplification and the other specific for amplification of sequences from Phytophthora spp. The plant primers amplified the desired amplicon size in the 29 plant species tested and did not interfere with amplification by the Phytophthora genus-specific primer pair. Using DNA from purified cultures, the Phytophthora genus-specific primer pair amplified a fragment diagnostic for the genus from all 45 Phytophthora spp. evaluated, although the efficiency of amplification was lower for P. lateralis and P. sojae than for the other species. The genus-specific primer pair did not amplify sequences from the 30 Pythium spp. tested or from 29 plant species, although occasional faint bands were observed for several additional plant species. With the exception of one plant species, the resulting amplicons were smaller than the Phytophthora genus-specific amplicon. The products of the first-round amplification were diluted and amplified with primer pairs nested within the genus-specific amplicon that were specific for either P. ramorum, P. nemorosa, or P. pseudosyringae. These species-specific primers amplified the target sequence from all isolates of the pathogens under evaluation; for P. ramorum, this included 24 isolates from California, Germany, and the Netherlands. Using purified pathogen DNA, the limit of detection for P. ramorum using this marker system was ≈2.0 fg of total DNA. However, when this DNA was spiked with DNA from healthy plant tissue extracted with a commercial miniprep procedure, the sensitivity of detection was reduced by 100- to 1,000-fold, depending on the plant species. This marker system was validated with DNA extracted from naturally infected plant samples collected from the field by comparing the sequence of the Phytophthora genus-specific amplicon, morphological identification of cultures recovered from the same lesions and, for P. ramorum, amplification with a previously published rDNA internal transcribed spacer species-specific primer pair. Results were compared and validated with three different brands of thermal cyclers in two different laboratories to provide information about how the described PCR assay performs under different laboratory conditions. The specificity of the Phytophthora genus-specific primers suggests that they will have utility for pathogen detection in other Phytophthora pathosystems. [ABSTRACT FROM AUTHOR]

The most common soilborne diseases affecting the strawberry industry in California include Verticillium wilt due to Verticillium dahliae, charcoal root rot due to Macrophomina phaseolina, and Fusarium wilt due to Fusarium oxysporum f. sp. fragariae. Detection of these pathogens in soil is an important facet of disease management and fumigation recommendations. Whereas the soil populations of both M. phaseolina and V. dahliae can be readily quantified with quantitative PCR (qPCR) assays using DNA extractions with 500 mg of soil, the single-cell nature of the F. oxysporum chlamydospore does not provide enough pathogen DNA from 500-mg extractions to be reliably quantified. Here, we describe an improved DNA extraction protocol from 10 to 15 g of soil that allows for the quantification of F. oxysporum f. sp. fragariae populations below 10 CFU/g. The relationship between results from the TaqMan qPCR assay and pathogen population density in soil was determined by using this extraction method in pathogen-free soils artificially infested with a hygromycin-resistant strain of F. oxysporum f. sp. fragariae to facilitate accurate colony counts when plated on a selective medium. Although the protocol was developed for F. oxysporum f. sp. fragariae, it is applicable for detection and quantification of other soilborne pathogens. [ABSTRACT FROM AUTHOR]

The effects of broccoli and lettuce rotations on population densities of Verticillium dahliae and Pythium spp. in soil and on strawberry (Fragaria x ananassa) growth, yield, and Verticillium wilt were evaluated in conventional and organic production systems in California for 2 years. Under both management systems, strawberry was planted after two successive crops of broccoli or lettuce. The control treatment in the conventional field was strawberry planted in soils fumigated with methyl bromide + chloropicrin. Preplant densities of V. dahliae and Pythiuni sp. did not differ in these fields. At the end of the second broccoli crop, V. dahliae densities in conventional plots had declined by 44% in both years. In contrast, after the second broccoli crop, densities in organic fields decreased 47% in 2000 and 25% in 2001. In general, there were no differences in V. dahliae inoculum densities in organic and conventional plots following lettuce rotations. After the second vegetable production cycle, population densities of V. dahliae in broccoli rotated organic (24 CFU/g of soil in 2000 and 27 CFU/g of soil in 2001) or conventional (23 CFU/g of soil in 2000 and 19 CFU/g of soil in 2001) fields were significantly lower than those in lettuce rotated organic (40 CFU/g of soil in 2000 and 42 CFU/g of soil in 2001) or conventional (39 CFU/g of soil in 2000 and 35 CFU/g of soil in 2001) fields. However, crop rotation treatments had no consistent effect on the inoculum densities of Pythium spp. Canopy diameters of strawberry plants grown in rotation with broccoli were not different from those in fumigated control plots, whereas those from lettuce plots were 10% smaller. Strawberry plant nutrient analysis showed that fertilizer inputs into organic or conventional production were not responsible for the observed differences in plant size. Increases in strawberry yields were not consistent between years. Verticillium wilt incidence on strawberry was 12 to 24% lower in fields rotated with broccoli compared with fields rotated with lettuce. Wilt severity on strawberry was 22 to 36% lower in fields rotated with broccoli compared with those rotated with lettuce. The strategy of using broccoli rotation coupled with postharvest incorporation of broccoli residue continues to show promise as a tool in the management of Verticillium wilt in both conventional and organic strawberry production systems. [ABSTRACT FROM AUTHOR]

Isolates of the Fusarium oxysporum species complex have been characterized as plant pathogens that commonly cause vascular wilt, stunting, and yellowing of the leaves in a variety of hosts. F. oxysporum species complex isolates have been grouped into formae speciales based on their ability to cause disease on a specific host. F. oxysporum f. sp. fragariae is the causal agent of Fusarium wilt of strawberry and has become a threat to production as fumigation practices have changed in California. F. oxysporum f. sp. fragariae is polyphyletic and limited genetic markers are available for its detection. In this study, next-generation sequencing and comparative genomics were used to identify a unique genetic locus that can detect all of the somatic compatibility groups of F. oxysporum f. sp. fragariae identified in California. This locus was used to develop a TaqMan quantitative polymerase chain reaction assay and an isothermal recombinase polymerase amplification (RPA) assay that have very high sensitivity and specificity for more than 180 different isolates of the pathogen tested. RPA assay results from multiple field samples were validated with pathogenicity tests of recovered isolates.

Macrophomina phaseolina is a broad-host-range fungus that shows some degree of host preference on strawberry, and causes symptoms that include crown rot and root rot. Recently, this pathogen has affected strawberry production as fumigation practices have changed, leaving many growers in California and around the world in need of accurate, rapid diagnostic tools for M. phaseolina in soil and infected plants. This study uses next-generation sequencing and comparative genomics to identify a locus that is unique to isolates within a main genotype shared by a majority of isolates that infect strawberry. This locus was used to develop a quantitative single-tube nested TaqMan polymerase chain reaction assay which is able to quantify as little as 2 to 3 microsclerotia/g of soil with 100% genotype specificity. An isothermal assay using recombinase polymerase amplification was developed from the same locus and has been validated on over 200 infected strawberry plants with a diagnostic sensitivity of 93% and a diagnostic specificity of 99%. Together, this work demonstrates the value of using new approaches to identify loci for detection and provides valuable diagnostic tools that can be used to monitor soil and strawberry plant samples for M. phaseolina.

Sudden oak death has been an emerging disease problem in coastal California and has caused significant losses in forest ecosystems in some regions of the state. The causal agent of this disease has been described as Phytophthora ramorum with two other less aggressive species, P. nemorosa and P. pseudosyringae, recovered from some symptomatic plants. The phylogenetic relationship of these species with other members of the genus was examined by sequence alignment of 667 bp of the mitochondrially-encoded cytochrome oxidase II gene and the nuclear encoded rDNA internal transcribed spacer region. P. ramorum was most closely related to P. hibernalis and P. lateralis in trees from both regions, although the specific relationship among species differed depending on the tree. In the cox II tree these species were on a single clade with P. lateralis basal to a group containing P. ramorum and P. hibernalis. On the maximum parsimony ITS tree P. ramorum was most closely affiliated with P. lateralis and in the same clade as P. hibernalis, but with maximum likelihood analysis P. ramorum was basal to a grouping of P. hibernalis and P. lateralis. While bootstrap support was strong for the grouping of these species together, it was not for determining the relationship among them. In contrast to the cox II tree, the clade containing these three species grouped with P. cryptogea, P. drechsleri, P. erythroseptica, and P. syringae in the ITS tree. Since the same isolates of these species were used for both the cox II and ITS sequence analysis, this difference in species grouping suggests either a differential rate of evolutionary divergence for these two regions, incorrect assumptions about alignment of ITS sequences, or different evolutionary histories of the regions under study. Analysis of combined cox II and ITS data sets gave trees where the relationships among these species were the same as for the ITS tree alone, although the results of the partition homogeneity test (P=0.072) suggest caution should be used in interpretation of this data. All analyses supported a close relationship between P. ilicis, P. nemorosa and P. pseudosyringae, although the analysis did not clarify the evolutionary relationships among these three species. Interestingly, these three species had a unique 6 bp deletion in the cox II gene just before the termination codon. While there was some similarity in phylogenetic grouping of these species and morphological characteristics, this was not consistent across all comparisons in the genus. Data would suggest that P. ramorum, P. nemorosa and P. pseudosyringae are phylogenetically distinct new species and not the result of interspecific hybridization.