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Shoot tip and single-node cutting explants of `Hamawy' and `El-Amar' apricot cultivars were initiated from forced shoots of field-grown, virus-free trees. Explants were cultured on Murashige & Skoog (MS) Nitsch & Nitsch and Anderson media. Different modifications of MS medium were also evaluated. Antioxidant pretreatment reduced phenolic compounds and decreased necrosis. Modified MS was the best medium for plantlets regeneration, with positive effectiveness of adenine sulfate addition to the modified MS. Shoot multiplication was best on 2.0 mg·L–1 BAP and 1.0 mg·L–1 thidiazuron (TDZ). Also, half-strength MS medium was superior for shoot elongation Surface coverage, 16 hours light/8 hours dark cycle, and 2.0 mg·L–1 IBA induced good rooting. Rooted plantlets were successfully acclimated ex vitro.

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In previous abstracts (HortScience 23:707;24:121), ABA when added throughout the in vitro production cycle, reversed the tissue culture-induced rejuvenation of the day neutral strawberry `Fern'. Compared to benzyl adenine (BA) proliferated plants, ABA treated tissue culture-produced plants flowered earlier and had more adult leaf patterns. In the present study, we analysed endogenous ABA concentrations in the apices and unexpanded leaves of BA treated tissue culture-propagated plants, selved seedlings and propagated adult runner tip plants at 3, 7 and 15 weeks ex vitro, after germination or after runner tip propagation. Using pentadeuterated standards and single ion monitoring, ABA concentrations in tissue culture produced and juvenile seedling plants were significantly lower than adult plants at 3 and 7 weeks. By 7 weeks, only the adult plants were flowering. At 15 weeks, no differences in ABA concentration were significant and all three types flowered.

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Hesperaloe parviflora is a useful xeric landscape plant. Two methods of shoot culture initiation were developed. Shoots were initiated indirectly through the use of callus derived from pieces of young inflorescences. Callus was initiated on modified Murashige and Skoog medium with 4.52 μM 2,4-dichlorophenoxyacetic acid and 2.32 μM kinetin. Callus produced shoots when placed on modified MS medium with 6.0 μM zeatin riboside. Direct initiation of shoots was also accomplished using the bottom four floral buds of young inflorescences. Buds were placed on modified MS media containing either benzylaminopurine or kinetin at 0.1, 1.0, or 10.0 μM or zeatin riboside at 6.0 μM. The most shoots were produced by the medium containing 6.0 μM zeatin riboside. Preliminary results indicate optimum shoot production with 2.0 to 4.0 μM BA or 6.0 μM zeatin riboside.Hesperaloe parviflora micro-shoots were rooted on one quarter strength MS medium and ex vitro.

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Experiments were conducted on tissue proliferation (TP) development and in vitro and ex vitro growth of tissues from plants with (TP+) and without TP (TP-). In 1993 the increase in TP in one-, two-, and three-yr-old `Holden' and `Besse Howells' was 3%, 52%. and 32% and 10%, 26% and 21%, respectively. No differential mortality was observed. Shoot tip cultures initated from TP+ and TP- `Montego' showed 10-12 mo were required for miniaturiziation and multiplication in TP- shoot tips and 4 mo in TP+ shoot tips. TP- cultures require 10 uM 2-iP for normal shoot proliferation; whereas TP+ cultures had to be transferred to hormone-free medium after 6 mo to maintain normal shoot morphology. Cutting propagation from TP- and TP+ plants older than 5 yr, showed persistence of morphological aberrations associated with TP+ plants.

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Cotyledons from developing embryos 6 to 8 weeks old of Liatris spicata (blazing star) were cultured on Murashige-Skoog (MS) medium containing 0, 0.4, 4.4, and 44.4 μ M benzyladenine (BA) or 0, 0.2, 2.2, and 22.2 μ M thidiazuron (TDZ) to induce adventitious shoot formation. The highest percent of cotyledons forming shoots with highest shoot counts was on medium containing 2.2 μ M TDZ. Vitreous shoots formed on medium with 22.2 μ M TDZ. Callus derived from cotyledons and cultured on medium containing 4.44 μ M BA or 2.2 μ M TDZ formed adventitious shoots with highest shoot counts on 4.44 μ M BA. Adventitious shoots derived from cotyledons and callus were rooted on MS medium with 5.0 μ Mindole-3-butyric acid, acclimatized and grown ex vitro. All micropropagated plants appeared similar to each other.

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Use of a liquid media during micropropagation has promoted improved proliferation and rooting response in several species. In this experiment, a double phase system (a combination of liquid and agar solidified medium) was applied to three cultivars of miniature roses (Rosa chinensis var. minima) to determine the effects on shoot quality and subsequent ex-vitro rooting. Applications of liquid media to the surface of agar solidified media were made at 0, 2, and 4 weeks. Evaluation via computerized image analysis after eight weeks of proliferation revealed equal or greater values for shoot length, area and weighted density (equivalent to fresh weight) for cultures receiving overlay, regardless of timing, compared to the solid media control. Additionally, application of a liquid overlay improved rooting response by up to 20% over the control and resulted in a tendency for a greater number of roots of greater length and area than the treatment without liquid media overlay.

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The effects of natural ventilation and CO2 enrichment during the rooting stage on the growth and the rates of photosynthesis and transpiration of in vitro cauliflower (Brassica oleracea L.) plantlets were investigated. In vitro plantlets were established in airtight or ventilated vessels with or without CO2 supplied (≈1200 μg·L-1) through gas permeable films attached to the vessel's cap for 15 days before transplanting ex vitro. Leaves generated in vitro in ventilated vessels had a higher photosynthetic rate than those produced in airtight vessels, which lead to greater leaf expansion and shoot and root dry matter accumulation during in vitro culture and acclimatization. Enhanced photosynthesis in leaves of ventilated plantlets was positively correlated with chlorophyll content. Increasing photosynthetically active radiation from 70 to 200 μmol·m-2·s-1 enhanced the growth of in vitro plantlets under ventilated conditions but it depressed photosynthesis of the leaves grown photomixotrophically with sugar and CO2 enrichment which might be due to the feedback inhibition caused by marked accumulations of sucrose and starch. Higher CO2 levels during in vitro culture enhanced photosynthesis under photoautotrophic conditions, but inhibited it under photomixotrophic conditions. Fifteen days after transplanting ex vitro, high photosynthetic ability and stomatal resistance to transpiratory water loss of ventilated plantlets in vitro had important contributions to rooting and acclimatization. Our findings show that the ventilated culture is effective for accelerating photoautotrophic growth of plantlets by increasing photosynthesis, suggesting that, especially for plantlets growing in vitro without sugar, CO2 enrichment may be necessary to enhance photosynthetic ability.

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A two-stage micropropagation system was devised for cranberries (Vaccinium macrocarpon Ait.). Shoot-tip explants taken from four cultivars of greenhouse-grown plants were placed on media composed of Anderson's major salts, Murashige and Skoog's (MS) minor salts and organics, plus various concentrations of 2iP, IBA, and GA3. In other experiments, explant source, salt formulations for media, and rooting treatments were studied. Optimal multiplication and shoot quality occurred when nodal explants taken from greenhouse-grown or micropropagated plants were placed on medium containing 150 μm 2iP, 1.0 μm IBA, and no GA3. Histological examination revealed that the initial response of nodes to culture is axillary bud proliferation, but adventitious shoot formation occurred after 4 to 6 weeks. Cultures that contained only axillary shoots were not evident unless low levels of 2iP were used, at which point only axillary buds present on the explants were released. Proliferated shoots could be rooted ex vitro without auxin treatment. Optimal rooting occurred under high-light conditions. Plants were transplanted to the field for comparison to conventionally propagated material. Chemical names used: gibberellic acid (GA3), N-(3-methyl-2-butenyl)-1H-purin-6-amine (2iP), 1H-indole-3-butanoic acid (IBA).

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Abstract

Foliar anatomical comparisons were made between in vitro-grown plantlets and greenhouse-grown plants of ‘Queen Elizabeth’ rose (Rosa sp.) using scanning and light microscopy. Each acuminate leaf apex and marginal serration had a terminal hydathode region composed of a glandular tip and a subterminal, adaxial group of sunken water pores. Leaf apices of greenhouse-grown plants had up to 35 water pores per hydathode, while cultured plantlets had < 20. Hydathodes of leaf serrations had up to 10 water pores in both sample groups. Water pores and stomata of plantlet leaves were open, while those of greenhouse-grown plants had smaller apertures or were completely closed. Internally, hydathodes were delimited by a bundle sheath extending below the vascular tissues and approaching the adaxial epidermis on each side of the water pore zone. Files of tracheary elements extended various distances into the leaf teeth. Small, irregularly shaped parenchyma cells (epithem) abutted on the xylem parenchyma cells, filling the space between the files of tracheary elements and the adaxial epidermis. The hydathodes of plantlet leaves were smaller with fewer water pores and reduced epithem than those of greenhouse-grown plants. Ex vitro guttation probably occurs as a result of increased water potential and high relative humidity when plantlets are transferred from culture to soil.

Open Access

Abstract

The hypothesis was tested that the growth of in vitro-propagated, nonrooted grape shoots (Vitis L. hybrid) directly after transfer from culture (ex vitro) is limited by photosynthetic C supply and that growth would be stimulated by CO2 enrichment (CDE). Plantlets were grown for 30 days in air with 350 or 1200 ppm (v/v) CO2 in humidified, flow-through chambers at 26°C. Destructive growth analyses were made at 0, 10, 20, and 30 days after transfer from culture to soil. CDE had no significant effect on total plant dry weight increase in the first 10 days. By 20 and 30 days, CDE-treated plants were 2 and 4 times greater in dry weight, respectively, than controls. Root growth was most improved by CDE, being almost 6 times greater than controls by 30 days. Leaf area per plant and root : shoot ratio were both doubled by CDE at 20 and 30 days. Since these results were under nonstress conditions, the use of CDE for growth stimulation needs to be evaluated under stress-hardening regimes.

Open Access