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The transfer of multigenic traits into tomato has been slow due to interspecific barriers (hybrid breakdown) found in the F2 of the Lycopersicon esculentum × L. pennellii cross (esc × pen), including blocks in normal reproductive development and nonfecundity. In a typical (esc × pen) F2 population, failure to flower and premeiotic blocks in pollen development occurred in 2% and 11% of the population, respectively. The remaining plants showed a mean of 37% stainable pollen. Twenty three percent of the F2 plants set seed, with an average of 4.5 seeds/fruit. An average of 33% of the stainable pollen from the 7 F2 plants with the highest stainable pollen measurements germinated in vitro, but only 4 of these 7 plants set seed. Thus, percent stainable pollen is not an adequate predictor of fecundity, and the non-fecundity in the F2Le plants must involve barriers occurring after pollen germination.
A method was developed which greatly reduces or eliminates each of the F2 barriers. The method and its efficacy on each of the aspects of hybrid breakdown will be discussed.
Protoplast isolation and culture protocols were developed for leaf tissue from 6 kenaf cultivars [Everglades 41 (E41), E71, Guatemala 4 (G4), G45, G51, and Tainung 1]. For protoplast isolation, the best combination of hydrolytic enzymes was cellulysin (1% w/v; Calbiochem) plus macerase (0.5% w/v; Calbiochem), with a 24 hour digestion at 30°C in the dark. Yields reached 7.2 (10)6 protoplasts/g leaf tissue. Protoplast viabilities ranged from 65% to 96%. Minor cultivar differences were observed related to protoplast yield, but all viability estimates were in an acceptable range. Greatest cell division frequencies and plating efficiencies were obtained when protoplasts were initially cultured in liquid medium at a density of 1.0 (10)5 protoplasts/ml. Electrofusion protocols were developed for kenaf protoplasts testing the range from 1200 to 3000 V/cm. A fusion voltage of 2000 V/cm yielded the highest fusion frequency and retained viability above 80%.
Abstract
Leaf anatomy and shoot growth of ‘Harrolds Red Delicious’ (‘HRD’) apple trees treated with succinic acid 2,2-dimethylhydrazide (Alar) were compared with those of an untreated compact mutant strain, ‘Starkrimson Delicious’ (‘SD’). Leaf anatomy of the untreated ‘HRD’ and ‘SD’ were not significantly different. Alar treatments of ‘HRD’ increased thickness of spongy parenchyma, palisade parenchyma, the length but not the no. of palisade cells and total leaf thickness. The site most affected by Alar was the palisade parenchyma. Alar also suppressed shoot growth rate and the treatment effect became noticeable 1 week after treatment. The shoot growth curves were quite similar for ‘SD’ and Alar-treated ‘HRD’.
The fine fescues are generally considered to be acid-tolerant compared to many other cool-season turfgrasses. However, there is a lack of documentation on aluminum tolerance of fine fescues at both the species and cultivar levels. A total of 58 genotypes belonging to five species or sub-species were screened under greenhouse conditions using solution culture, sand culture, and acid Tatum subsoil. This soil had 69% exchangeable Al and a pH of 4.4. An Al concentration of 640 μM and a pH 4.0 were used in solution screening and sand screening. Differences in Al tolerance were identified at both species and cultivar levels based on relative growth. The genotypes with endophyte infection generally exhibited greater Al tolerance than endophyte-free genotypes. The results indicate that fine fescues vary in Al tolerance and there is potential to improve Al tolerance with breeding and to refine management recommendations for fine fescues regarding soil pH.
Lipase activity was studied during endodormancy in low-chilling-requiring `Anna' and high-chilling-requiring `Northern Spy' apples (Malus domestica Borkh.). Lipase activity greatly increased in bud axes when the chilling requirement of buds was almost satisfied regardless of the absolute chilling needed. Lipase activity greatly increased in `Anna' after 400 chill units (CU) and in `Northern Spy' after 2600 CU. This corresponded with an increase in budbreak at 22 to 24C. The increase in lipase activity also coincided with the release of water in buds from the bound to the free form. We propose that lipase(s) activity is an integral part of breaking dormancy and that lipase participates in causing changes in membrane lipid composition that coincides with releasing water into the free form.
Cucurbit leaf crumple geminivirus (CuLCrV) is transmitted by sweet-potato whitefly (Bemisia tabaci) biotype B (SPWF-B) and occurs on cucurbits in Arizona, California, Texas, and Mexico. This virus is identical to Cucurbit leaf curl virus, and their symptoms are similar to Squash leaf curl virus on squash (Cucurbita sp.) and Melonleaf curl virus on melon (Cucumis melo L.). Melon has been reported to be either susceptible to CuLCrV, or to have the ability to recover from infection. Twenty-three melon cultigens were inoculated with CuLCrV in greenhouse tests using SPWF-B. Eighteen of the cultigens tested were highly susceptible to CuLCrV (≥60% infected plants) and generally exhibited pronounced CuLCrV symptoms: `Amarillo', `Edisto 47', `Esteem', `Fuyu 3', `Impac', `Moscatel Grande', `Negro', `Perlita', PI 234607, PI 236355, PI 414723, `PMR 5', `Seminole', `Sol Dorado', `Sol Real', `Top Mark', `Vedrantais', and WMR 29. Five cultigens were resistant to CuLCrV (<40% infected plants that exhibited restricted, mild symptoms): MR-1, PI 124111, PI 124112, PI 179901, and PI 313970. Symptoms abated with time in both groups although infected plants remained positive for the virus. Ten of the cultigens (`Edisto 47', `Fuyu 3', `Impac', MR-1, PI 124112, PI 313970, PI 414723, `PMR 5', `Top Mark', and WMR 29) were included in field tests in 2003 and 2004 that were naturally infected with CuLCrV. With the exception of PI 414723, the greenhouse and field data were consistent for reaction to CuLCrV.
As part of a program to develop transgenic peach (Prunus persica L. Batsch) cultivars with resistance to Prunus necrotic ringspot virus (PNRSV), we are testing a system for measuring virus in peach shoot cultures. Micrografting in vitro is used for inoculation and slot-blot hybridization, with a digoxigenin (DIG)-labeled cRNA probe complementary to the 5′ open reading frame (ORF) of PNRSV RNA 3, for detection. In this study, we investigated whether infected shoots maintain virus infection over long periods of culture at 4 °C and if PNRSV-infected `Suncrest' shoot cultures can serve as graft bases to transmit virus equally well into cultivars Nemaguard, Springcrest, and Suncrest. The results of RNA hybridization analysis showed that virus was present in extracts of leaf samples from 2-year-old PNRSV-infected `Suncrest' shoots that had been subjected to varying lengths of incubation at 4 °C in the dark, suggesting that infected shoots can be maintained for repeated use. Rates of graft success were higher in heterografts between `Suncrest' bases and tips of `Springcrest' or `Nemaguard' than in autografts between `Suncrest' and `Suncrest', and there was equal efficacy of graft inoculation from `Suncrest' into these three cultivars.
Pepper (Capsicum annuum L. `Early Calwonder') leaf disks were vacuum-infiltrated in distilled water (control), anisomycin, aurintricarboxylic acid, cycloheximide, ethionine, norvanine, or puromycin to determine whether protein synthesis inhibitors blocked high-temperature acclimation. After infiltration, one-half of the leaf disks were placed in an incubator at 24C as a control, and the other half were kept in a water bath at 38C for 2 h to induce acclimation. Test tubes containing the disks then were placed in a water bath at 50.5C for 0, 1, 5, 10, 15, 25, 35, or 50 minutes. Thermotolerance was evaluated using electrolyte leakage. High-temperature acclimation was blocked in all six protein synthesis-inhibitor treatments. Only control disks infiltrated with distilled water acclimated. It seems that protein synthesis is required for high-temperature acclimation in bell pepper leaves.
Household detergents were evaluated in field studies on fresh-market tomato (Lycopersicon esculentum Mill.) for insecticidal and phytotoxic effects. Laboratory bioassays were used to examine the toxicity of a household liquid dish detergent on small nymphs of silverleaf whitefly, Bemisia argentifolii Bellows and Perring. The detergents tested proved to be more toxic to whitefly nymphs than the commercial insecticidal soap. Detergent treatments were applied to tomato with a commercial high pressure hydraulic sprayer at 0%, 1%, 2%, 4%, and 8% (by volume) initially and at 0%, 0.25%, 0.5%, 1.0%, and 2.0% (by volume) in subsequent tests. As detergent rate, frequency of application, or both increased, plant dry weight accumulation and fruit yield decreased. Applying detergent also increased time to fruit maturity. A once-a-week application of 0.25% to 0.5% detergent initially applied 2 weeks after transplanting alleviated phytotoxicity and yield reduction problems.
Avocado (Persea americana Mill.) tissues contain high levels of the seven-carbon (C7) ketosugar mannoheptulose and its polyol form, perseitol. Radiolabeling of intact leaves of `Hass' avocado on `Duke 7' rootstock indicated that both perseitol and mannoheptulose are not only primary products of photosynthetic CO2 fixation but are also exported in the phloem. In cell-free extracts from mature source leaves, formation of the C7 backbone occurred by condensation of a three-carbon metabolite (dihydroxyacetone-P) with a four-carbon metabolite (erythrose-4-P) to form sedoheptulose-1,7-bis-P, followed by isomerization to a phosphorylated d-mannoheptulose derivative. A transketolase reaction was also observed which converted five-carbon metabolites (ribose-5-P and xylulose-5-P) to form the C7 metabolite, sedoheptulose-7-P, but this compound was not metabolized further to mannoheptulose. This suggests that C7 sugars are formed from the Calvin Cycle, not oxidative pentose phosphate pathway, reactions in avocado leaves. In avocado fruit, C7 sugars were present in substantial quantities and the normal ripening processes (fruit softening, ethylene production, and climacteric respiration rise), which occurs several days after the fruit is picked, did not occur until levels of C7 sugars dropped below an apparent threshold concentration of ≈20 mg·g-1 fresh weight. The effect of picking could be mimicked by girdling the fruit stalks, which resulted in ripening on the tree. Again, ripening followed a decline in C7 sugars to below an apparent threshold level. Taken together, these data indicate that the C7 sugars play important roles in carbon allocation processes in the avocado tree, including a possible novel role as phloem-mobile ripening inhibitors.