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Armillaria root rot (ARR) of peach caused by the soil-borne basidiomycete fungus Armillaria tabescens is causing premature decline and mortality of peach trees on most southeastern U.S. peach farms. Soil inoculum may be present both in former peach orchard sites and on sites that were once in hardwood forest. The fungus is protected under the bark of dead root pieces and may survive up to 100 years at various depths in the soil profile. No commercially available rootstocks are resistant to ARR. Since 2002, we have embarked on a multipronged strategy to develop control options to combat ARR. First, we have two replicated trials on commercial grower replant sites with a history of ARR. Trial 1 compares four preplant fumigation treatments (none, Telone II, methyl bromide, and Enzone), three rootstocks (Lovell, Halford, and Guardian) and preplant root dips with endomycorrhizal fungi. Trial 2 compares the use of raised beds, root collar excavation and preplant root dips. Both trials examine long-term productivity and tree survival. Second, we are examining the use of systemic fungicide injection into infected trees to protect trees around infection foci. Third, we are trying to develop a genetically modified ARR-resistant rootstock. We have inserted the gene encoding the gastrodia antifungal protein (GAFP—a low molecular weight lectin that binds mannose and chitin) from a Chinese orchid into tobacco (model herbaceous system) and plum (model Prunus system). GAFP has antifungal activity against several basidiomycete root rot pathogens. Pathogenicity tests with transformed tobacco plants show enhanced tolerance to several root rot pathogens when compared to nontransformed plants. Transformed plums are being multiplied for pathogenicity tests.
Rooted cuttings of `Halford' and `Redhaven' peaches [Prunus persica (L.) Batsch] and `Stanley' (Prunus domestica L.) and `Marianna 2624' (P. cerasifera × P. munsoniana) plums were planted in soil containing ≈38 tomato ringspot virus-(TomRSV) infested nematodes (Xiphinema americanum sensu lato Cobb) per 100 cc. Test- and control-plant sap extracts were made from root and leaf tissues after 10, 22, and 34 weeks. Aliquots of these samples were assayed by mechanical inoculation to Chenopodium quinoa Willd. Total nucleic-acid extracts prepared from the remainder of each sample were analyzed by dot blot hybridization using a cRNA probe for TomRSV. The bioassay identified one `Stanley' and two `Redhaven' infected plants. Hybridization results indicated that two of two `Stanley', three of three `Halford', five of five `Redhaven', and zero of six `Marianna 2624' were infected. Our results demonstrate the sensitivity of molecular hybridization for TomRSV detection in Prunus and substantiate the TomRSV resistance of `Marianna 2624'.
The evergreen (EVG) peach, first described in Mexico, was used as a parent with deciduous (DE) peaches to develop F1 and F2 hybrid populations in Mexico, Florida, Georgia, and West Virginia. F1 trees were DE and F2 plants segregated 3 DE: 1 EVG. In West Virginia, the most temperate location, the heterozygous class could be distinguished in the first few years of growth by late leaf abscission in the fall. Segregation ratios suggest that the EVG trait is controlled by a single gene, evg, the EVG state being homozygous recessive. Evergreen trees were characterized by insensitivity of shoot tips to daylength and failure of terminal growth to cease growth until killed by low temperature. Lateral buds of EVG trees went dormant in the fall. Deep supercooling occurred in both EVG and DE trees, but it appeared later in EVG trees, was of shorter duration, and occurred to a lesser extent. Evergreen germplasm may be useful in developing peach cultivars for frost-free subtropic and tropical areas. It also presents a useful system for studying dormancy and cold hardiness.
Abstract
Sample sizes for detection of differences of flower bud survival in peach and nectarine [Prunus persica (L.) Batsch] were chosen on the basis of theoretical confidence intervals (Cl) and least detectable differences (LDD) for the binomial distribution. Theoretical Cl and LDD for 1000-bud samples were comparable to Cl and Duncan's multiple range test separation computed from an analysis of variance for 1000 buds, based upon 10 replicates of 100 buds. Variability in survival was a function of eultivar, height of bud in canopy, and bud type. Variability may be minimized by sampling a given bud type (single, double, distal) at >1.5 m above ground level.
Tobacco (Nicotiana tabacum cv Wisconsin 38) leaf discs were transformed with the disarmed Agrobacterium tumefaciens strain EHA101 carrying the Rol C gene from A. rhizogenes (Oono et al., Jpn. J. Genet. 62:501-505, 1987), NPT II and GUS. Shoots that regenerated on kanamycin-containing medium were confirmed transgenic through GUS assays, Southern analyses and transmission of foreign genes through the sexual cycle. Transgenic plants were as short as half the height of control plants, earlier flowering by up to 35 days, had smaller leaves, smaller seed capsules, fewer seeds, smaller flowers and reduced pollen viability. The number of seed capsules, leaf number and root density were similar between transgenic and control plants. Transgenic clones varied in the expression of the Rol C gene and transgenic plants similar or only slightly different from controls were identified. Transformation with the Rol C gene presents a potentially useful method of genetically modifying horticultural crops, particularly for flowering date, height, and leaf and flower size.
`Wisconsin 38' tobacco (Nicotiana tabacum L.) leaf discs were transformed with the disarmed Agrobacterium tumefaciens strain EHA101 carrying the rolC gene from A. rhizogenes (Oono et al., 1987) and NPT II and GUS genes. Shoots that regenerated on kanamycin-containing medium were confirmed as transgenic through GUS assays, polymerase chain reaction (PCR), Southern blot analyses, and transmission of the foreign genes through the sexual cycle. Transgenic plants were as short as half the height of control plants; were earlier flowering by up to 35 days; and had smaller leaves, shorter internodes, smaller seed capsules, fewer seeds, smaller flowers, and reduced pollen viability. The number of seed capsules, leaf number, and specific root length were similar between transgenic and control plants. Transgenic clones varied in the expression of the rolC-induced growth alterations as did the first generation of seedlings from these clones. Such differences suggested the potential for selecting for different levels of expression. Transformation with the rolC gene presents a potentially useful method of genetically modifying horticultural crops, particularly for flowering date, height, and leaf and flower size. Chemical names used: neomycin phosphotransferase (NPTII), β-glucuronidase (GUS).
Peach [Prunus persica (L.) Batsch.] is considered the best genetically characterized species of the genus Prunus. We therefore used it as a model in our study of the genome organization in Prunus by means of restriction fragment length polymorphisms (RPLPs). Initial results indicated that 60% of cloned DNA sequences examined occur at low copy number within the peach genome. After selecting and examining these sequences, polymorphisms sufficient for RPLP mapping were found. We determined that ≫33% of our cDNA clones and 20% of our genomic clones detected RPLPs among peach cultivars. Analysis of RPLP segregation in two families, both of which segregate for known morphological characters, revealed segregation in 12 RFLP markers for one family and 16 for the other. Although we have not detected linkage between RFLP and morphological markers, preliminary analyses indicate possible linkage between two RPLP markers.
We have constructed a genetic linkage map of peach consisting of RFLP, RAPD, and morphological markers, based on 78 F2 individuals derived from the self-fertilization of four F1 individuals originating from a cross between `New Jersey Pillar' and KV 77119. This progeny set was chosen because parental genotypes exhibit variation in canopy shape, fruit flesh color, and flower petal color, size, and number. The segregation of 81 markers comprised of RFLP, RAPD and morphological loci was analyzed. Low copy genomic and cDNA probes were used in the RFLP analysis. The current genetic map for the WV family contains 57 markers assigned to 9 linkage groups, which cover 520 cM of the peach nuclear genome. The average distance between two adjacent markers was 9 cM. Linkage was detected between Pillar (Pi) and double flowers (Dl). RFLP markers loosely linked to Pi, flesh color (Y), and white flower (W) loci were found. Twenty-four markers remain unassigned.
Transgenic grape plants were regenerated from somatic embryos derived from leaves of in vitro-grown plants of `Thompson Seedless' grape (Vitis vinifera L.) plants. Somatic embryos were either exposed directly to engineered Agrobacterium tumefaciens or they were bombarded twice with 1-μm gold particles and then exposed to A. tumefaciens. Somatic embryos were transformed with either the lytic peptide Shiva-1 gene or the tomato ringspot virus (TomRSV) coat protein (CP) gene. After cocultivation, secondary embryos proliferated on Emershad/Ramming proliferation (ERP) medium for 6 weeks before selection on ERP medium containing 40 μg·mL-1 kanamycin (kan). Transgenic embryos were identified after 3 to 5 months under selection and allowed to germinate and develop into rooted plants on woody plant medium containing 1 μm 6-benzylaminopurine, 1.5% sucrose, 0.3% activated charcoal, and 0.75% agar. Integration of the foreign genes into these grapevines was verified by growth in the presence of kanamycin (kan), positive β-glucuronidase (GUS) and polymerase chain-reaction (PCR) assays, and Southern analysis.
Abstract
‘Bounty’ peach [Prunus persica (L.) Batsch] was released because of its large fruit size, excellent flavor (as judged by us), and productivity, particularly under dry soil conditions of eastern Texas. Its ability to produce fruit of uniform maturity throughout the canopy makes it especially suitable for once-over harvesting. ‘Bounty’ has outstanding potential as a mid-season fresh-market peach for the south-central United States, particularly Texas, and is suggested for trial in the mid-Atlantic and eastern United States.