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- Author or Editor: Lan Liu x
- HortScience x
A scale-up process of lettuce (Lactuca sativa L.) suspension culture in a 2-liter bioreactor was investigated. Factors that influenced cell growth and differentiation, including foaming, the wall effect (inoculum adhering onto the vessel wall above the medium level), aeration, and dissolved oxygen (DO), were tested. The wall effect resulted in severe inoculum loss (10%) in 24 hours. Inoculum loss significantly decreased shoot regeneration. The wall effect was caused by two factors: 1) foaming caused by the interaction between air bubbles and inoculum, and 2) the bubbles produced by aeration. Foaming could be prevented by sieving the inoculum through a 400-pm screen filter and then rinsing the inoculum thoroughly with distilled water to remove single cells, cell debris, and the contents of broken cells. The wall effect caused by air bubbles could be prevented by putting a 150-μm screen column in the center of the bioreactor to isolate the aeration area from the inoculum. After the wall effect was removed, shoot regeneration in the bioreactor increased significantly to a level similar to that in 125-ml flasks at an aeration rate of 1 to 2 vvm (liters air/liters medium per rein). DO for this shoot regeneration level was ≈ 70% to 80%of saturation at the end of bioreactor culture.
An efficient method for the regeneration of shoots directly from cell suspensions of three commercial cultivars of lettuce (Lactuca sativa L. cv. Great Lakes 659-700, Salad Bowl, and Prize Head) is described. Cell suspensions were prepared by osterizing cotyledon-derived callus for 60 seconds. The effects of callus quality, light intensity, carbohydrate type and concentration, auxins, and cytokinins on cell growth and differentiation in the suspension culture were examined. Among these factors, callus quality and carbohydrates were the most critical. The optimal medium for regeneration of shoots in suspension culture was SH (Schenk and Hilderbrandt) basal medium containing 1000 mg myo -inositol/liter, 1.5% glucose, 0.44 μm BA, and 0.54 μm NAA. The pH of the medium was adjusted to 5.8. Under such condition, hundreds of shoots could be produced from 50 to 55 mg (dry wt) of cell aggregates within 2 weeks. Chemical names used: α-] naphthaleneacetic acid (NAA); indole3-acetic acid (IAA); 6-benzylaminopurine (BA).
A bioreactor was used to establish a scale-up system for somatic embryogenesis in `Scarlet' carrot (Daucus carota L.). At a cell density of 1–2 × 106 cells/ml, mature and germinating embryos could be observed within 4 to 5 weeks. As cell density exceeded 2 × 106 cells/ml, the culture turned darker yellow, and embryo development was inhibited. Cell densities below 106 cells/ml resulted in abnormal embryos. Bioreactor design had a critical impact on somatic embryogenesis due to various types and the strength of shear forces generated. In this study, an air-lift bioreactor was selected from three different types (spinner flask, screen column bioreactor, and air lift) because it resulted in the highest biomass production and somatic embryogenesis. Foaming was eliminated by preculture of embryogenic cells in flasks; cells were then sieved on a 60-μm polyester screen and thoroughly rinsed with distilled water before being transferred to the bioreactor. Such preculture for at least 10 days significantly increased the regeneration of somatic embryos. During somatic embryogenesis, dissolved O2 concentrations decreased to 33% of saturation, and then increased up to 80% when embryo development approached maturity and mature embryos germinated. Bioreactor-cultured embryos germinated with relatively short cotyledons and long roots, whereas flask-cultured embryos germinated with relatively long cotyledons and short roots.
Dendrobium wardianum is a key ornamental plant and a valuable traditional Chinese medicine. This research aimed to find the optimal protocol for in vitro inducement of polyploidy in D. wardianum by treating protocorms with colchicine (an antimitotic agent). The experiment consisted of two series of treatments. For the first treatment, the protocorms were subjected to colchicine concentrations of 25, 75, 125, 250, and 500 μM (weight/volume) for 6, 12, and 24 hours. For the second treatment, protocorms were cultivated in culture medium with colchicine (25, 75, 125, and 250 μM) for 30 days. A total of 18 polyploids were confirmed by chromosome counts and anatomical parameters. Polyploids had broad, dark green leaves with increased stem lengths compared with those of diploids. The optimal protocol for these two methods consisted of soaking in 250 μM of colchicine solution for 12 hours, resulting in inductivity of 26%, and cultivating in 75 μM for 30 days, resulting in a mutation rate of 34%. A comparison of these two protocols showed that the latter one induced more stable polyploids, but that the survival rate was slightly lower. The survival and induced mutation rates of these plants were significantly influenced by the colchicine concentration and exposure time. Higher concentrations for longer periods of time resulted in greater mortality rates and longer-lasting side effects. The protocol involving a solid medium and colchicine is worth considering. It will be intriguing to examine this methodology for the induction of stable polyploids of other orchid species.
Rosaceous crops (e.g., almond, apple, apricot, caneberry, cherry, pear, peach, plum, rose, and strawberry) contribute to human health and well-being and collectively constitute the economic backbone of numerous North American rural communities. We conducted a survey of U.S. and Canadian rosaceous fruit crop breeders to assess priority setting in their programs, sources of information for setting priorities, and challenges in making technical and management decisions. Input from producers and consumers was most important in establishing breeding program targets, although respondents’ direct interaction with consumers was not frequent. Breeding targets and management decisions were mostly associated with the breeder’s type of organization, scope and range of crops, and intended use of the crop (fresh, processed, or both).
Hydrogen sulfide (H2S) was recently recognized as an endogenous gaseous molecule involved in seed germination, root organogenesis, abiotic stress tolerance, guard cell movement, and delay of senescence in plants. In the present study, we show that H2S participates in the regulation of postharvest ripening and senescence in fresh-cut kiwifruit, Actinidia deliciosa. Fumigation of fresh-cut kiwifruit with the H2S donor sodium hydrosulfide (NaHS) solution prolonged kiwifruit storage time and alleviated senescence and tissue softening in a dose-dependent manner at an optimal concentration of 1.0 mmol·L−1 NaHS. H2S treatment maintained higher levels of reducing sugars, soluble proteins, free amino acids, ascorbate, and chlorophyll and lowered carotenoid levels. H2S treatment also significantly decreased the contents of malondialdehyde (MDA), hydrogen peroxide (H2O2) and superoxide anion (•O2 −) during fruit storage compared with water controls. Furthermore, the activities of guaiacol peroxidase (POD), catalase (CAT), ascorbate peroxidase (APX), and glutathione reductase (GR) were increased by H2S treatment, whereas the activity of lipoxygenase (LOX) was decreased compared with untreated controls. Taken together, these results suggest that H2S is involved in prolonging postharvest shelf life and plays an antioxidative role in fresh-cut kiwifruit.
Hydrogen sulfide (H2S) has been shown to be a gaseous molecule in the regulation of many processes in plants such as abiotic stress tolerance, root organogenesis, stomatal movement, and postharvest fruit senescence. We studied the role of H2S in the regulation of senescence and fungal decay in fresh-cut sweetpotato (Ipomoea batatas L., cv. Xushu 18) roots. H2S donor sodium hydrosulfide (NaHS) alleviated senescence in fresh-cut sweetpotato root tissue in a dose-dependent manner with the optimal concentration of 2.0 mmol·L−1 NaHS solution. At the optimal concentration of 2.0 mmol·L−1 NaHS, H2S fumigation maintained higher levels of reducing sugar in sweetpotato fresh-cut root. H2S treatment also significantly increased the activities of guaiacol peroxidase (POD) and decreased those of polyphenol oxidase (PPO) in sweetpotato during storage. Further investigation showed that H2S treatment maintained a lower level of lipoxygenase (LOX) activity compared with water control. Consistently, the accumulation of malondialdehyde (MDA) was reduced in H2S-treated groups. Three fungal pathogens, Rhizopus nigricans, Mucor rouxianus, and Geotrichum candidum, were isolated from sweetpotato tissue infected with black rot or soft rot. H2S fumigation at 1 to 2.5 mmol·L−1 NaHS resulted in effective inhibition of the three fungi when grown on medium. When the three fungi were inoculated on the surface of sweetpotato slices, H2S fumigation greatly reduced the percentage of fungal infection. In conclusion, these data suggest that H2S effectively alleviated the senescence and decay in sweetpotato slices and might be developed into a novel fungicide for reduction of black rot or soft rot in sweetpotato.
Hydrogen sulfide (H2S) has been proven to be a multifunctional signaling molecule in plants. In this study, we attempted to explore the effects of H2S on the climacteric fruit tomato during postharvest storage. H2S fumigation for 1 d was found to delay the peel color transition from green to red and decreased fruit firmness induced by ethylene. Further investigation showed that H2S fumigation downregulated the activities and gene expressions of cell wall–degrading enzymes pectin lyase (PL), polygalacturonase (PG), and cellulase. Furthermore, H2S fumigation downregulated the expression of ethylene biosynthesis genes SlACS2 and SlACS3. Ethylene treatment for 1 d was found to induce the expression of SlACO1, SlACO3, and SlACO4 genes, whereas the increase was significantly inhibited by H2S combined with ethylene. Furthermore, H2S decreased the transcript accumulation of ethylene receptor genes SlETR5 and SlETR6 and ethylene transcription factors SlCRF2 and SlERF2. The correlation analysis suggested that the fruit firmness was negatively correlated with ethylene biosynthesis and signaling pathway. The current study showed that exogenous H2S could inhibit the synthesis of endogenous ethylene and regulate ethylene signal transduction, thereby delaying fruit softening and the ripening process of tomato fruit during postharvest storage.