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Thomas S.C. Li and W.R. Schroeder

Sea buckthorn (Hippophae rhamnoides L.) is a multipurpose, hardy, deciduous shrub, an ideal plant for soil erosion control, land reclamation, wildlife habitat enhancement, and farmstead protection. It has high nutritional and medicinal values for humans. The majority of sea buckthorn research has been conducted in Asia and Europe. It is a promising new crop for North America, and recently it has attracted considerable attention by researchers, producers, and industry.

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Yin-ming Li and W.H. Gabelman

Sixty highly homozygous tomato (Lycopersicon esculentum Mill.) strains, some selected from previous studies and some collected from known low-Ca regions, were screened under a low-Ca culture system (10 mg of Ca per plant). Four strains were selected to represent the extremes for Ca efficiency and used as parents to create a series of F1, F2, and backcross generations for inheritance studies of Ca use under low-Ca stress. Based on total plant dry weight, additive and dominance gene effects were most important for the efficiency of Ca use. Maternal control of efficiency in Ca use was not observed. Estimates of broad-sense heritability ranged from 63% to 79% for total play dry weight. Narrow sense heritabilities, determined in only two of the families, were 47% to 49$ and 68% to 75%.

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Kimberly J. Felcher, D.S. Douches, W.W. Kirk, R. Hammerschmidt, and W. Li

Research was done to determine if enhanced resistance to potato (Solanum tuberosum L.) late blight could be obtained by combining host plant resistance and engineered resistance. Late blight susceptible cultivars, Atlantic, and Spunta and the partially resistant cultivar Libertas were transformed with a fungal glucose oxidase gene, resulting in lines which ranged in transgene copy number from 1 to 8. Glucose oxidase enzyme activity ranged from 0.00 to 96.74×10-5 units/mg plant tissue. There was no correlation between copy number and level of transgene mRNA, level of transgene mRNA and enzyme activity, or between level of enzyme activity and disease resistance. Field and growth chamber evaluation of late blight response demonstrated little to no effect of the glucose oxidase transgene in either late blight susceptible or partially late blight resistant cultivars. However, enzyme activity levels were much lower than levels reported in previous research, which may account for the lack of effect of glucose oxidase against Phytophthora infestans. Twenty-one percent of the transgenic lines were phenotypically off-type compared to nontransgenic controls. Most of the off-type transgenic lines (four out of seven) were derived from `Libertas'. Because several off-type lines did not express the glucose oxidase protein, this phenomenon could not be attributed solely to the glucose oxidase transgene. Based on these results, transgenic lines produced for this study do not increase resistance to P. infestans even in combination with moderate host plant resistance. However, production of greater numbers of transgenic lines with the current construct or, production of transgenic lines in which a different constitutive promoter drives the expression of the glucose oxidase gene might result in greater disease resistance. However, the usefulness of any small increase in resistance would need to be evaluated against the time and cost required for development of transgenic potato cultivars and the potential for off-type tubers and plants.

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Jing-fen Chen, Paul H. Li, and David W. Davis

Exposure of young pepper plants to chilling temperatures delays the development of terminal flower buds to flowering during post-stress growth. Degree of adverse influence depends on chilling intensity, exposure duration and varietal sensitivity. `Ma Belle' pepper plants were grown in a greenhouse (GH) during winter months on the St. Paul campus, No supplemental lighting was provided. When plants were at the 2- to 3-leaf stage, they were foliar sprayed with mefluidide (Technical grade) at 0, 5, 10 and 15 ppm. One day after treatment, some plants were transferred from GH to a cold room (3° ∼4°C day/night) with 12-h photoperiod. Treatad plants remaining in the GH served as the control. Plants were chilled for 1, 2, 4 and 6 days and then brought back to the GH for post-stress growth and development observation. Treated and untreated plants grown in the GH showed no difference in days to flowering, and reached 50% flowering at about 62 days after treatment. When untreated plants were chilled for 1,2,4 and 6 days, they showed a delay of 8, 18, 30 and 34 days, respectively, to flowering, If not killed, as compared to the control The long delay to flowering was due to the injury of the terminal flower buds. After 4 and 6 days of chilling, most terminal flower buds were killed. However, when plants were treated with mefluidide and subsequently chilled days to flowering were significantly shortened. A difference of 10-12 days was observed between chilled untreated plants and chilled treated plants. Concentrations of 5 to 15 ppm were equally effective in protection against chilling.

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Jing-fen Chen, Paul H. Li, and David W. Davis

Exposure of young pepper plants to chilling temperatures delays the development of terminal flower buds to flowering during post-stress growth. Degree of adverse influence depends on chilling intensity, exposure duration and varietal sensitivity. `Ma Belle' pepper plants were grown in a greenhouse (GH) during winter months on the St. Paul campus, No supplemental lighting was provided. When plants were at the 2- to 3-leaf stage, they were foliar sprayed with mefluidide (Technical grade) at 0, 5, 10 and 15 ppm. One day after treatment, some plants were transferred from GH to a cold room (3° ∼4°C day/night) with 12-h photoperiod. Treatad plants remaining in the GH served as the control. Plants were chilled for 1, 2, 4 and 6 days and then brought back to the GH for post-stress growth and development observation. Treated and untreated plants grown in the GH showed no difference in days to flowering, and reached 50% flowering at about 62 days after treatment. When untreated plants were chilled for 1,2,4 and 6 days, they showed a delay of 8, 18, 30 and 34 days, respectively, to flowering, If not killed, as compared to the control The long delay to flowering was due to the injury of the terminal flower buds. After 4 and 6 days of chilling, most terminal flower buds were killed. However, when plants were treated with mefluidide and subsequently chilled days to flowering were significantly shortened. A difference of 10-12 days was observed between chilled untreated plants and chilled treated plants. Concentrations of 5 to 15 ppm were equally effective in protection against chilling.

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D.S. Douches, W. Li, K. Zarka, J. Coombs, W. Pett, E. Grafius, and T. El-Nasr

The potato tuber moth (Phthorimaea operculella Zeller) is the primary insect pest of cultivated potato (Solanum tuberosum L.) in tropical and subtropical regions, causing both foliar and tuber damage. In contrast, the Colorado potato beetle (Leptinotarsa decemlineata Say) is the most important insect pest in the northern potato production latitudes. The codon-modified Bacillus thuringiensis Bt-cry5 gene (revised nomenclature cry1IaI), specifically toxic to Lepidoptera and Coleoptera, was transformed into cultivar Spunta using an Agrobacterium vector to provide resistance to both potato tuber moth and Colorado potato beetle. The Bt-cry5 gene was placed downstream from the constitutive CaMV35S promoter. Two transgenic 'Spunta' clones, G2 and G3, produced high levels of mortality in first instars of potato tuber moth in detached-leaf bioassays (80% to 83% mortality), laboratory tuber tests (100% mortality), and field trials in Egypt (99% to 100% undamaged tubers). Reduced feeding by Colorado potato beetle first instars was also observed in detached-leaf bioassays (80% to 90% reduction). Field trials in the United States demonstrated that the horticultural performance of the two transgenic lines was comparable to 'Spunta'. These Bt-cry5 transgenic potato plants with high potato tuber moth resistance have value in integrated pest management programs.

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D.S. Douches, T.J. Kisha, J.J. Coombs, W. Li, W.L. Pett, and E.J. Grafius

The Colorado potato beetle, Leptinotarsa decemlineata (Say), is the most serious insect pest of potatoes throughout the eastern and north central United States. Host plant resistance to the Colorado potato beetle has been identified in wild Solanum species and Bt-transgenic potato lines. Detached-leaf bioassays (72 h) were conducted on insecticide-resistant, first instar Colorado potato beetles to study the effectiveness of individual and combined host plant resistance traits in potato. Potato lines tested include non-transgenic cultivars (`Russet Burbank', `Lemhi Russet', and `Spunta'), a line with glandular trichomes (NYL235-4), a line with high foliar leptines (USDA8380-1), and transgenic lines expressing either codon-modified Bt-cry3A or Bt-cry5 (Bt-cry1Ia1). Bt-cry3A transgenic lines, foliar leptine line, and foliar leptine lines with Bt-cry5 had reduced feeding compared to non-transgenic cultivars. Glandular trichome lines and glandular trichome lines with Bt-cry5 did not reduce feeding in this no-choice feeding study. Some Bt-cry5 transgenic lines, using either the constitutive promoters CaMV35s or (ocs)3mas (Gelvin super promoter), were moderately effective in reducing larval feeding. Feeding on Bt-cry5 transgenic lines with the tuber-specific patatin promoter was not significantly different than or greater than feeding on the susceptible cultivars. Mortality of first instars was highest when fed on the Bt-cry3A lines (68% to 70%) and intermediate (38%) on the Bt-cry5 `Spunta' line SPG3 where the gus reporter gene was not included in the gene construct. Host plant resistance from foliar leptines is a candidate mechanism to pyramid with either Bt-cry3A or Bt-cry5 expression in potato foliage against Colorado potato beetle. Without multiple sources of host plant resistance, long-term sustainability is questionable for a highly adaptable insect like the Colorado potato beetle.

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Wei Li, Rongcai Yuan, Jacqueline K. Burns, L.W. Timmer, and Kuang-Ren Chung

Colletotrichum acutatum J. H. Simmonds infects citrus flower petals, causing brownish lesions, young fruit drop, production of persistent calyces, and leaf distortion. This suggests that hormones may be involved in symptom development. To identify the types of hormones, cDNA clones encoding proteins related to ethylene and jasmonate (JA) biosynthesis, indole-3-acetic acid (IAA) regulation, cell-wall modification, signal transduction, or fruit ripening were used to examine differential gene expressions in calamondin (Citrus madurensis Lour) and/or `Valencia' sweet orange (Citrus sinensis Osbeck) after C. acutatum infection. Northern-blot analyses revealed that the genes encoding 1-aminocyclopropane-1-carboxylate (ACC) oxidase and 12-oxophytodienoate required for ethylene and JA biosynthesis, respectively, were highly up-regulated in both citrus species. Both gene transcripts increased markedly in petals, young fruit and stigmas, but not in calyces. The transcripts of the genes encoding IAA glucose transferase and auxin-responsive GH3-like protein, but not IAA amino acid hydrolyase, also markedly increased in both species 5 days after inoculation. The expansin and chitinase genes were slightly up-regulated, whereas the senescence-induced nuclease and ß-galactosidase genes were down-regulated in calamondin. No differential expression of transcripts was detected for the genes encoding expansin, polygalacturonase, and serine-threonine kinase in sweet orange. As compared to the water controls, infection of C. acutatum increased ethylene and IAA levels by 3- and 140-fold. In contrast, abscisic acid (ABA) levels were not significantly changed. Collectively, the results indicate that infection by C. acutatum of citrus flowers triggered differential gene expressions, mainly associated with IAA, ethylene, and JA production and regulation, and increased hormone concentrations, consistent with the hypothesis of the involvement of phytohormones in postbloom fruit drop.

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Li Lu, Kirk W. Pomper, Jeremiah D. Lowe, and Sheri B. Crabtree

Pawpaw (Asimina triloba) is a tree fruit native to eastern North America, which is in the early stages of domestication. Most early 20th century pawpaw cultivars have been lost; however, recent cultivar releases and potential new releases may have enhanced genetic diversity. The objective of this study was to compare the genetic variation exhibited among older and new pawpaw cultivars and Kentucky State University (KSU) advanced selections using simple sequence repeat (SSR) markers. Polymorphic microsatellite marker analysis was conducted with nine older pawpaw cultivars, six recently released PawPaw Foundation (PPF) cultivars, and nine KSU advanced selections. Using 18 microsatellite loci, a total of 179 alleles were amplified in the set of 24 genotypes. The major allele frequency (0.13 to 0.96), number of genotypes (two to 23), and allele size (96 to 341 bp) varied greatly by locus. Eighteen loci were highly polymorphic, as indicated by high expected heterozygosity (He = 0.71) and observed heterozygosity (Ho = 0.65) values as well as high polymorphism information content (polymorphism information content = 0.69). The dinucleotide SSR (GA and CA motifs) loci were more polymorphic than trinucleotide (ATG and AAT motifs) SSRs. The PPF cultivars and KSU advanced selections were more closely grouped genetically than with older cultivars. Older cultivars displayed the greatest genetic diversity (Ho = 0.69). The pawpaw cultivar base of older and PPF cultivars does appear to be genetically diverse. However, KSU advanced selections contain unique pawpaw germplasm that should enhance the genetic base of cultivars if these selections are released to the public.

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Fumiomi Takeda, Gerard Krewer, Changying Li, Daniel MacLean, and James W. Olmstead

Northern highbush (NH) blueberry (Vaccinium corymbosum) and southern highbush (SH) blueberry (V. corymbosum hybrids) have fruit that vary in firmness. The SH fruit is mostly hand harvested for the fresh market. Hand harvesting is labor-intensive requiring more than 500 hours/acre. Rabbiteye blueberry (V. virgatum) tends to have firmer fruit skin than that of NH blueberry and has been mostly machine harvested for the processing industry. Sparkleberry (V. arboreum) has very firm fruit. With the challenges of labor availability, efforts are under way to produce more marketable fruit using machine harvesting. This could require changing the design of harvesting machine and plant architecture, and the development of cultivars with fruit that will bruise less after impact with hard surfaces of machines. The objectives of this study were to determine the fruit quality of machine-harvested SH blueberry, analyze the effect of drop height and padding the contact surface on fruit quality, investigate the effect of crown restriction on ground loss, and determine the effect of plant size on machine harvestability. The fruit of ‘Farthing’, ‘Scintilla’, ‘Sweetcrisp’, and several selections were either hand harvested or machine harvested and assessed during postharvest storage for bruise damage and softening. Machine harvesting contributed to bruise damage in the fruit and softening in storage. The fruit of firm-textured SH blueberry (‘Farthing’, ‘Sweetcrisp’, and selection FL 05-528) was firmer than that of ‘Scintilla’ after 1 week in cold storage. Fruit drop tests from a height of 20 and 40 inches on a plastic surface showed that ‘Scintilla’ was more susceptible to bruising than that of firm-textured ‘Farthing’ and ‘Sweetcrisp’. When the contact surface was cushioned with a foam sheet, bruise incidence was significantly reduced in all SH blueberry used in the study. Also, the fruit dropped 40 inches developed more bruise damage than those dropped 20 inches. Ground loss during machine harvesting was reduced from 24% to 17% by modifying the rabbiteye blueberry plant architecture. Further modifications to harvesting machines and plant architecture are necessary to improve the quality of machine-harvested SH and rabbiteye blueberry fruit and the overall efficiency of blueberry (Vaccinium species and hybrids) harvesting machines.