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Vanhoutte's spiraea has been propagated in vitro using explants from softwood growth of dormant stems forced in a solution containing 200 mg/l 8-hydroxyquinoline citrate (8-HQC) and 2% sucrose (Yang and Read, 1989). Objectives to further utilize this system were to determine the feasibility of applying plant growth regulators (PGR) via the forcing solution to softwood growth from forced dormant stems and to study the resulting influence on in vitro culture. BA and GA3 were placed in the forcing solution at various concentrations, including a zero PGR control. Explants were cultured on Linsmaier and Skoog (LS) medium containing zero PGR or different amounts of BA or thidiazuron (TDZ) or combinations of BA and IAA. Control explants placed on LS medium supplemented with 5uM BA with or without 1 or 5uM IAA, or with 0.5 or 0.75 uM TDZ alone produced the best shoot proliferation. BA in the forcing solution stimulated micropropagation, while GA3 caused less proliferation than explants from control solutions. Forcing solutions containing PGR are useful for manipulating responses of plant tissues cultured in vitro and for studying PGR influence on woody plant physiology.
American chestnut (Castanea dentata) is one of the United States' most valuable resources for its nuts and timber. Many scientists are exploring genetic transformation techniques to improve chestnut blight resistance in addition to conventional breeding. In vitro shoot production must be first obtained and optimized in order to establish an efficient transformation system. Although shoot proliferation has been achieved, chestnut is still considered difficult for tissue culture with poor rooting. Therefore, this research has focused on improving rooting ability of micropropagated chestnut shoots. In vitro shoot production was established and maintained in WPM supplemented with 0.1 mg/l BA, 3% sucrose, and 0.7% agar with the pH adjusted to 5.8. The shoots were then transferred to rooting medium containing the same components as for shoot proliferation plus an auxin at various concentrations. Right after placing shoots onto rooting medium, a very thin layer (5 ml) of the same auxin (diluted) was added to provide a quick stimulation of rooting. Detailed discussion will be presented.
Chestnut (Castanea spp.) is considered difficult to micropropagate. The timing for harvesting explant materials from forced stems is critical, although many factors need to be considered for successful micropropagation. Previous research with spirea and five-leaf aralia demonstrated that forcing solution techniques extended the availability of high-quality explant material, thus expediting micropropagation. However, preliminary research illustrated that chestnut is very difficult to force and the new forced softwood growth is very short-lived, which made micropropagation difficult. It was found that, at about 7 days from budbreak, the forced chestnut softwood growth (about 2 cm long) served as the best explant material. If longer than this timing window, the new growth would die. If shorter, the explants had a high contamination rate, exudation of purported phenolic compounds, and explants would not regenerate. Shoot proliferation and callus regeneration were achieved by culturing good-quality explants on Woody Plant Medium supplemented with 0.1 mg BA/liter. The new shoots grew vigorously in vitro with apparent normal morphology.
The goal of this study was to expedite galax seed germination in vitro. Galax seeds were collected from Yancey County, N.C., at an elevation of about 1100 m. Aseptic cultures were established using the tiny rust-colored seeds. In vitro seed germination was achieved under different pH conditions (4.2, 5.0, and 5.8). Seeds cultured in the medium with pH 4.2 tended to germinate early with a better rate than those cultured with a higher pH of 5.0 or 5.8 at the very beginning. Gradually, seeds from media with pH 5.0 and 5.8 caught up in germination. Eventually, seeds from all pH treatments produced a very similar germination rate. Attempts to use the matted and scaly rhizomes and very tender new growth as explant materials to establish aseptic cultures were not successful, due to severe contamination. However, our observations suggested that the very tender new growth could be a good source of explants once the optimum sterilization time is established.
To overcome the limitations of traditional propagation, this research was initiated to develop an alternative means for efficient production of Alexandrian laurel (Danae racemosa L. Moench). An in vitro propagation protocol has been developed for Danae racemosa L. Moench using seeds as a source of material for culture initiation. Seedlings were produced after seeds were cultured for 3 month on MS () medium. Shoot multiplication occurred on MS medium with or without 6-benzylaminopurine (BAP) with 100% multiplication percentage. However, shoot number was significantly increased from an average of 2.8 to more than six with the addition of 5 or 25 μM BAP. Among two indole-3-butyric acid (IBA) treatments tested for rooting of seedlings, incorporation of 5 μM IBA in MS medium significantly increased rooting percentage to 86.4% compared with 71.2% without IBA. The greatest number of roots (three) was produced by 5-minute IBA pulse. However, both IBA treatments significantly reduced root length. The longest root (12.8 mm) was observed on MS medium without any IBA treatment and the shortest (6.1 mm) was produced by IBA pulse. In vitro-propagated plantlets grew well after transfer to a substrate of peat and pine bark (1:1) in the greenhouse. No morphological variation was observed.
This study investigated antimicrobial effects of guava products on the survival and growth of Escherichia coli O157:H7 in liquid medium. Seven strains of E. coli O157:H7 (944, 380, E0019, F4546, H1730, Cider, 9727) were tested. These strains were maintained in BHI broth. Guava fruits were sliced into small pieces and blended using a blender. Guava juice and leaves were then extracted using three solvents: water, methanol and hexane. Fruit extracts were dissolved in 10 ml BHI broth tubes to make a fruit solution of 5% (w/v). E. coli O157:H7 was inoculated into fruit solutions at 2 log cfu/mL. After incubation at 37 °C for 24 h, samples were serially diluted 10 folds. The proper diluent was spread-plated on TSA in duplicate. After incubation at 35 °C for 24 h, viable cell counts were obtained. The experiment was replicated three times in a randomized complete-block design. Results demonstrated that guava products (fruit, juice, and leaf extracts) significantly reduced survival and growth of the tested foodborne pathogen strains. Water extract showed the highest antimicrobial activity, followed by methanol and hexane. These results indicate guava extracts are a potential antimicrobial agent to ensure food safety.
Black cohosh (Actaea racemosa L.), a medicinal herb commonly used in herbal supplements for the treatment of various ailments, is a perennial herb that grows naturally under shade conditions in temperate forest regions. This project studied the growth and rhizome yield of Black cohosh under shade conditions of 0%, 40%, 60%, and 80% in a high tunnel (9.1 m wide × 29.3 m long) on the North Carolina Agricultural and Technical University Farm. Seed rhizomes were planted in raised beds incorporated with 9070 kg/acre compost and preplant fertilizer on 29 May 2016. There was one row per bed, with in-row spacing at 45.7 cm, and one drip line per bed for irrigation. Fertigation was done weekly through the drip tapes with Multi-K 13–0–46 (27.2 kg N/acre) during the growing season. Beds were mulched after sprouting. Growth data of fully mature plants were collected on canopy width and length, total number of stems per plant, stem diameter, and length/height; and rhizome fresh and dry weight. Data were analyzed at the 0.05 level of significance. Plant canopy, stem diameter, and length/height were significantly greater in 40% shade (average, 504.7 × 472.6 mm, 3.7 mm, and 135.9 mm, respectively) than in other shade conditions, with the smallest sizes in 0% shade (average, 255.8 × 255.7 mm, 2.1 mm, and 95.4 mm, respectively). There were no significant differences between the 60% and 80% shade conditions in plant canopy, stem diameter, and length/height. However, the total number of stems per plant (4.9) in 0% shade was significantly more than those in other shade conditions, with the least of stems per plant (2.9) in 80% shade. Rhizome fresh and dry weight per plant were the greatest (164.6 and 48.1 g, respectively) in 40% shade, and the least (77.8 and 22.5 g, respectively) in 0% shade. The results indicate that optimum growing conditions for Black cohosh was in 40% shade with a Daily light integral (DLI) between 15 and 0 mol/m2/day, and a day- and nighttime temperature difference between 8.3 and 2.7 °C.
Plug seedlings of delphinium (Delphinium elatum L. `Guardian') were planted on 19 Nov. 2004 in four production systems (harvest lugs, lay-flat bags, pots, and polystyrene trays). Production systems were randomized in a Latin-square design with four replications of each system. Each treatment plot was 0.7 × 1.1 m. Planting density was 31 plants/m2. The harvest lugs were 55 × 37 × 16 cm. The lay-flat bags were 114 × 30 × 3 cm. The pots were 25 cm bulb pans. The polystyrene trays were 67 × 34 × 5 cm and contained 32 square cells. All of the containers were filled with the same tobacco germination media. The plants in the harvest lugs, lay-flat bags and pots were irrigated daily with 150 mg N/kg from 20N–4.4P–16.6K. The plants in the polystyrene trays were floated on a solution of 150 mg N/kg from 20N–4.4P–16.6K. Float solutions were monitored and adjusted weekly for volume and fertilizer concentration. Individual stems were harvested at the one-third bloom stage of development. The final harvest was on 1 Apr. 2005. Fewer stems were harvested from float trays and lay-flat bags than from pots and harvest lugs. The stems harvested from float tray plots were shorter than those from the other three systems. Stem fresh weight from greatest to least was lay-flat bags, harvest lugs, pots, and float trays. Stem dry weight was less for float trays than the other three systems.
This research was initiated to study different culture media and plant growth regulators for their influences on callus initiation and production, with a research goal of developing an efficient in vitro callus regeneration protocol for guava (Psidium guajava L.). Guava is an important tropical fruit species that is rich in vitamins and vitamin precursors, minerals, organic acids, and pectins. Seventy-nine phytochemicals provide guava with many unique properties and actions, including anti-microbial, astringent, bactericidal, cicatrizant, emmenagogue, hypoglycemic, laxative, nutritive, and spasmolytic. Different concentrations of various plant growth regulators (PGR), such as 6-benzyladenine (BA), kinetin, or 2,4-dichlorophenoxyacetic acid (2,4-D), and naphthaleneacetic acid (NAA) were added to basic Murashige and Skoog (MS) and woody plant medium (WPM) and tested for their influences. Differences in callus initiation and morphology were noticed between MS and WPM, and among PGR concentration treatments.
Plug seedlings of Tagetes erecta L. `Gold Coin Mix' were planted in four production systems (harvest lugs, lay-flat bags, pots, and polystyrene trays) on 5 May 2005. Production systems were randomized in a Latin-square design with four replications of each system. Each treatment plot was 0.7 m × 1.1 m. Planting density was 31 plants/m2. The harvest lugs were 55 cm × 37 cm × 16 cm. The lay-flat bags were 114 cm × 30 cm × 3 cm. The pots were 25-cm bulb pans. The polystyrene trays were 67 × 34 × 5 cm and contained 32 square cells. All of the containers were filled with the same tobacco germination media. The plants in the harvest lugs, lay-flat bags, and pots were irrigated on alternate days with 150 mg·kg-1 N from 20N–4.4P–16.6K. The plants in the polystyrene trays were floated on a solution of 150 mg·kg-1 N from 20N–4.4P–16.6K. Float solutions were monitored and adjusted weekly for volume and fertilizer concentration. Individual stems were harvested at the appropriate stage of development for market. The fresh weight, stem length, and dry weight of individual stems were recorded. The rate of growth and maturation differed between production systems and locations in the greenhouse. Detailed results will be presented.