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  • Author or Editor: Z. Li x
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Industrial-scale cultivation of plant cells for valuable product recovery (e.g. natural pigments, pharmaceutical compounds) can only be considered commercially-feasible when a fully-automated, predictable bioprocess is achieved. Automation of cell selection, quantification, and sorting procedures, and pinpointing of optimal microenvironmental regimes can be approached via machine vision. Macroscopic staging of Ajuga reptans callus masses (ranging between 2-6 g FW) permitted simultaneous rapid capture of top and side views. Area data used in a linear regression model yielded a reliable, non-destructive estimate of fresh mass. Suspension culture images from the same cell line were microscopically imaged at 4x (with an inverted microscope). Using color machine vision, the HSI (hue-saturation-intensity) coordinates were used to successfully separate pigmented cells and aggregates from non-pigmented cells, aggregates, and background debris. Time-course sampling of a routine suspension culture consistently allowed pigmented cells to be detected, and intensity could be correlated with the degree of pigmentation as verified using spectrophotometer analysis of parallel samples.

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Plukenetia volubilis Linneo, a tropical recurrent woody oilseed plant native to South America, was successfully introduced in China. A field experiment was conducted to determine the effect of the dry-season foliar sprays once every 2 weeks with 50 μm water or five different plant growth regulators (PGRs) viz., gibberellic acid (GA3), kinetin (KIN), indole-3-acetic acid (IAA), abscisic acid (ABA), and salicylic acid (SA) on the growth and yield of P. volubilis plants in Xishuangbanna, southwest China. Results showed that PGRs affected the leaf stomatal conductance (g S) and water-use efficiency (WUEi), rather than the net photosynthetic rate (PN). The phenological development of P. volubilis plants, including the time of flowering and maturity, and the dynamic pattern of fruit ripening, was not altered by PGR treatments. ABA and SA resulted in highest fruit set, seed oil content, and total fruit or seed oil yield, whereas GA3, IAA, and KIN were effective in increasing seed size. The nonstructural carbohydrates (NSC) are related to subsequent abscission or retention of the developing fruit, which was indicated by the positive relationship between carbohydrate concentration and fruit set across PGR treatments. The positive influences of PGRs on the total fruit yield (increased 4.3% to 15.2%) and total seed oil yield (increased 4.9% to 24.9%) per unit area throughout a growing season were found when compared with the control, depending to a great extent on the balance between vegetative and reproductive growth during the reproductive stage. This study suggests that PGRs, especially for ABA and SA, can become a valuable tool for promoting the seed oil yield of P. volubilis plants while maintaining high seed quality in the field.

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Fertilization is among the most important factors influencing fruit quality of citrus. Effects of Individual element such as N, P, or K on fruit quality have been well-documented. Much less has been done on the interactions of N, P, and K in relation to citrus fruit quality. A field experiment was conducted from 1994 to 1999 in a commercial grove on a Riviera fine sand (Loamy, siliceous, hyperthermic Arenic Glossaqualf) to investigate the effects of fertilizer rates and sources on fruit quality of 26-year-old `White Marsh' grapefruit trees (Citrus paradisi Macfad.) on Sour Orange rootstock (Citrus aurantium Lush). Fertilizer was applied as water-soluble dry granular broadcast (three applications/year) at N rates of 0, 56, 112, 168, 224, and 336 kg/ha per year using a N;P:K blend (1.0:0.17:1.0). There was a quadratic relationship between fruit weight or peel thickness and fertilizer rates. Fruit weight per piece increased with fertilizer rates from 0 to 168 kg N/ha per year, but decreased from 168 to 336 kg N/ha per year. Fruit size was small at zero or low fertilizer rates due to nutrient deficiencies. Large fruit sizes of `White Marsh' grapefruit in the sandy soil were achieved at fertilizer rate around 168 kg N/ha per year. Increasing fertilizer application rates higher than 168 kg/ha per year greatly increased the number of fruit per tree, but decreased the size of fruit. Peel thickness, which is related to the fruit size, declines at higher fertilizer rates. Increase in fertilizer rate from 0 to 336 kg N/ha per year increased solids content and fruit acid concentration of the grapefruit. Fertilization rate effect on fruit Brix concentration was more complicated. Brix concentration was not affected by increasing fertilizer rates from 0 to 168 kg N/ha-per year, but was increased at higher fertilization rates (168 to 336 kg N/ha per year). As a result, the Brix/acid ratio was, in general, decreased by increasing fertilizer rates.

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Pierce's disease (PD), caused by the xylem-limited bacterium Xylella fastidiosa, is endemic to the coastal plain of the southeastern United States. Although native southern grapevines are tolerant to X. fastidiosa, all varieties of Vitisvinifera grown in the region will succumb to PD. Genetic transformation to add disease resistance genes, while not disturbing desirable phenotypic characters, holds promise for expanding the southeastern U.S. grape industry by allowing use of established fruit and wine varieties. We utilize embryogenic cell cultures and Agrobacterium strain EHA105 to refine transformation systems for Vitis species and hybrids. V. vinifera`Thompson Seedless' is employed as a model variety to test various transgenes for disease resistance, since as many as 150 independent transgenic plant lines routinely are produced from 1 g of embryogenic culture material. Transgenic plants are stringently screened for PD resistance in greenhouses by mechanical inoculation with X. fastidiosa. Transgenic plants are compared with both susceptible and resistant control plants by assessing typical PD symptom development and by assaying bacterial populations in xylem sap over time. Using these procedures, nine putative PD resistance genes have been inserted into grapevine and over 900 unique transgenic lines have been evaluated. A range of susceptible-to-resistant responses has been catalogued. Thus far, the best construct for PD resistance contains a grape codon-optimized hybrid lytic peptide gene in a high-performance bi-directional 35S promoter complex. Certain transgenic plant lines containing this construct exhibit better resistance than that of resistant control vines.

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