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A pollen grain undergoes a series of biochemical changes during germination. The technique of cylindrical internal reflectance FTIR was used to examine spectral frequencies associated with respiration, lipid and protein structure, polysaccharide content, and phosphate-containing metabolizes in pollen from pecan, blue spruce, cattail, and pine. Samples of both pollen and germination medium were analyzed at timed intervals. A microscopic evaluation of percent germination was also made at each sampling time. Preliminary analyses indicate that changes in respiration occur as evidenced by the presence of gaseous CO2, and that quantitative changes in lipid and protein occur. FTIR spectroscopy provides a noninvasive method to directly and quantitatively measure metabolic changes associated with pollen germination.
Genetically engineered (GE), virus-resistant papaya cultivars in Hawaii are easily identified by a colorimetric assay for the β-glucuronidase (GUS) marker transgene. We used GUS to track pollen movement from a central 1-acre plot of gynodioecious GE `Rainbow' plants into seeds on surrounding border rows of non-GE `Sunrise' papaya. GUS evidence of cross-pollination occurred in 70% of female plants (43% of assayed seeds), compared with only 13% of the predominantly self-pollinating hermaphrodite plants (7% of seeds) segregating in the gynodioecious `Sunrise' border rows. The percentage of GUS+ seeds in border row plants showed a weak negative correlation (r = –0.32) with distance from the nearest GE tree (30 m maximum). In a non-GE papaya field located less than a mile downwind from the `Rainbow' source, no evidence of GUS was found in 1000 assayed seeds. In a separate study, the origin of GUS+ seed discovered in papaya fruits from an organic farm was investigated. Leaf GUS assays revealed that 70% of trees were GE, indicating that the grower had planted GE seed. The impact of pollen drift from GE trees in the same field was determined by screening seed samples from 20 non-GE hermaphrodites for GUS expression. Only three hermaphrodites (15%) showed GUS+ seeds, at low levels ranging from 3% to 6% of contaminated samples. These data indicate that the major source of GE contamination in organic fields is seeds of unverified origin, rather than pollen drift from neighboring GE fields. Organic growers are advised to: 1) plant only seed that is known to be non-GE, preferably obtained by manual self-pollination of selected non-GE hermaphrodites; 2) avoid open-pollinated seed; and 3) grow only hermaphrodite (self-pollinating) trees, removing any female or male plants from production fields.
A comprehensive educational program focusing on water quality protection was developed for the Oklahoma nursery industry. The program focused on best management practices to limit pesticides and nutrients in irrigation runoff and on capture and recycle technology as a pollution prevention strategy. Key professionals from the departments of entomology and plant pathology, biosystems and agricultural engineering, and horticulture formed a multidisciplinary team within the Oklahoma Cooperative Extension Service (OCES). During 1998, water quality workshops were conducted on-site throughout Oklahoma at leading nursery operations. These workshops were designed to highlight best management practices (BMPs) that were already in place as a foundation on which to implement additional BMPs with the assistance of the OCES team. Training workshops were augmented by written publications, by web-based information, and by videotape instruction. These provided for ongoing education beyond the formal grant period. The written materials included a water quality handbook for nurseries and a fact sheet on capturing and recycling irrigation runoff. The water quality handbook was also made available on the web and a website on disease management for nurseries using recycling irrigation was provided. The water quality video, highlighting successful growers, was designed to show aspects of both best management practices and capture and recycle technology. Results of these 3-year extension efforts will be discussed.
Application of four DNA fingerprint probes to avocado (Persea americana Mill.) resulted in identification of various cultivars, characterization of the three avocado races, and a genetic analysis of family structure. Genomic DNA from 14 cultivars was probed with four DNA fingerprint probes. Three of the probes gave well-resolved bands. The individual-specific patterns obtained for each cultivar validate the use of this technique for definitive cultivar characterization, with the probability of obtaining a similar pattern for two different cultivars being 2 × 10-9. DNA mixes representing either Mexican, Guatemalan, or West-Indian avocado races were hybridized with the DNA fingerprint probes, and a band pattern characteristic for each race was obtained. Progeny of a cross between the cultivars Ettinger and Pinkerton were analyzed. Their DNA fingerprints revealed one pair of linked bands and another band allelic to one of them. The application of these observations to identification, evolutionary studies, and breeding is discussed.
DNA fingerprint information was used for identification of mango (Mangifera indica L.) cultivars for genetic relatedness analysis of20 mango cultivars and for genetic analysis of a family structure. Genomic DNA was extracted from young leaves, digested with Hind III or Dra I, and hybridized with 10 different DNA probes. Jeffreys' minisatellite probe 33.6 was the most useful, resulting in well-resolved bands representing highly polymorphic loci. Specific patterns were obtained for each cultivar. The probability of obtaining a similar pattern for two different cultivars was 9.4 × 10-6. Based on DNA fingerprint information, genetic distances between 20 mango cultivars were evaluated and an evolutionary tree was established. Analysis of DNA fingerprint band patterns of 12 progeny resulting from a cross between `Tommy Atkins' and `Keitt' mango revealed neither linked nor allelic bands. Application of the reported results for identification, genetic analyses, and mango breeding is discussed.
Mother plants from strawberry (Fragaria ×ananassa Duch.) genotypes susceptible to and resistant to verticillium (Verticillium dahliae Kleb.) were inoculated with this pathogen in a high elevation nursery. The infection rate for mother plants was 77.3% ± 7.5% and 80.7% ± 5.4% for resistant and susceptible genotypes, respectively. Conversely, the percentage of runner plants identified as infected by postharvest petiole assay that were produced by these inoculated mother plants differed significantly (P < 0.01) between sets of genotypes, 25.1% ± 3.7% and 59.8% ± 5.3% for resistant and susceptible genotypes, respectively. The percentage of runner plants from inoculated mothers that eventually collapsed in the fruit production field was larger than the percentage identified as infected by petiole assay for susceptible genotypes (68.9% ± 4.1%), and substantially less than the pre-plant infected fraction for resistant genotypes (3.5% ± 1.4%). Yield for runner plants from inoculated mothers was reduced by 73% to 75% for susceptible genotypes, and by 7% to 15% for resistant genotypes. The percentage of runner plants identified as infected in the nursery was correlated with the percentage of plants that collapsed in the fruiting field (r = 0.91, P < 0.01) and with yield in infested plots (r = -0.79, P < 0.01). Most of the effect of this disease was expressed as plant collapse, but the presence of yield reductions larger than the rate of plant collapse demonstrated substantial sub-lethal effects as well.
Strawberry cultivars Selva and Camarosa (Fragaria ×ananassa Duch.) were grown at a high elevation nursery in soil that was either naturally infested with Verticillium dahliae or was rendered pathogen-free through preplant fumigation with 2 methyl bromide: 1 chloropicrin (wt/wt) at 392 kg·ha-1. Plants grown in fumigated soil were inoculated with a conidial suspension of V. dahliae, prior to establishment. Just prior to harvest, plants were rated for disease based on symptoms of Verticillium wilt. At the same time, petiole samples were taken from mother plants and each of three generations of runner plants, along with the stolons subtending each of the sampled runner plants. Petioles and stolons were cultured to assay for the presence of V. dahliae, and scored as either infected or not infected. The experiment was conducted in each of two successive years, and the following conclusions were supported by results obtained in both years. First, symptoms of Verticillium wilt on mother plants of both cultivars were highly correlated with recovery of V. dahliae from petioles, but runner plants were consistently free of symptoms even though they were often infected. Second, runner plants sustained lower infection frequencies than mother plants, with the differences being significant in most cases. Lastly, infection of runner plants was due, at least in part, to transfer of inoculum from infected mother plants; in some cases this appears to have been the exclusive mode of infection.
Organic methods for managing striped cucumber beetles (Acalymma vittatum) and spotted (Diabrotica undecimpunctata) cucumber beetles were examined in the production of watermelon (Citrullus lanatus) and muskmelon (Cucumis melo) using sticky traps to monitor beetle populations. In 2002, the numbers of trapped striped and total (striped + spotted) cucumber beetles were significantly (P ≤ 0.05) reduced by the combined use of three companion plants thought to repel cucumber beetles [radish (Raphanus sativus), tansy (Tanacetum vulgare), and nasturtium (Tropaeolum spp.)] or by the combined use of three companion plants known to attract beneficial insects [buckwheat (Fagopyrum esculentum), cowpeas (Vigna unguiculata), and sweetclover (Melilotus officinalis)]. In 2003 and 2004, the single companion plant treatment consisted of the combined use of radish and buckwheat. In 2003, use of aluminum-coated plastic mulch (Al-plastic) or companion plants significantly increased muskmelon yields and vine cover, while significantly reducing numbers of trapped striped, spotted, and total cucumber beetles. The use of pyrethrin insecticide did not significantly affect muskmelon yields or vine cover. In 2004, the beneficial effects of companion plant and Al-plastic treatments on muskmelon yields and vine cover were also significant and similar to those in 2003; however, these treatments only affected early season numbers of trapped beetles. The use of rowcovers significantly increased muskmelon yields and vine cover in 2003 and 2004 and did not affect beetle populations after rowcover removal. It was concluded that use of companion plants and Al-plastic increased muskmelon yields and vine cover while reducing populations of cucumber beetles, particularly striped cucumber beetles. The use of rowcovers also increased muskmelon yields and vine cover.