In an attempt to improve the micropropagation protocol for lingonberry (Vaccinium vitis-idaea L.) developed at the Centre, two lingonberry clones were compared for in vitro shoot proliferation on two different media supplemented with varying levels of thidiazuron (TDZ). TDZ supported proliferation at low concentrations (0.1 to 1 μm) but inhibited shoot elongation. However, usable shoots were obtained within 4 weeks by transferring shoot cluster to medium containing 1 μm zeatin. Genotypes differed significantly with respect to multiplication rate with `EL1' producing the most shoots per explant. In both genotypes, shoot proliferation was greatly influenced by explant orientation. Changing the orientation of explants from vertically upright to horizontal increased axillary shoot number, but decreased shoot height and leaf number per shoot. Proliferated shoots were rooted on a 2 peat: 1 perlite (v/v) medium, and the plantlets were acclimatized and eventually established in the greenhouse.
Sachiko Matsubara and Hegazi H. Hegazi
Callus initiation and growth and plantlet regeneration were studied using eight cultivars of Raphanus sativus L., including six Japanese radishes, one Chinese and one small `Comet' radish. The basal medium was composed of Murashige and Skoog inorganic salts, 2.0 mg myo-inositol/liter, 0.5 mg each of nicotinic acid and pyridoxine·HCl/liter, and 0.1 mg thiamine·HCl/liter, 30 g sucrose and 2 g Gelrite/liter. High callus yields were obtained on basal medium containing (mg·liter-1) 0.1 2,4-D and 1.0 BA for two Japanese radishes and 0.1 NAA and 1.0 kinetin for `Comet' radish. Shoots were regenerated from callus by subculturing on basal medium containing 0.1 or 1.0 mg BA/liter and then transferring to basal medium. Rooting occurred on basal medium. Although callus was obtained in all eight cultivars, shoots and plantlets were regenerated only from `Moriguchi', `Nerima Shirinaga', and `Comet'. Chemical names used: 2-(l-naphthyl) acetic acid (NAA); N-(phenylmethyl)-lH-purine-6-amine (BA); 2,4-dichlorophenoxy acetic acid (2,4-D); 6-(furfurylamino)purine (kinetin).
Makoto Nakatani, Masaru Tanaka, and Masaru Yoshinaga
A late-storage root-forming mutant (`KM95-A68') of sweetpotato [Ipomoea batatas (L.) Poir.] was characterized to clarify the genetic and physiological mechanisms of storage root formation. This mutant originated from a somaclonal mutation of `Kokei No. 14'. Storage roots of `KM95-A68' are rare and, when formed, develop 2 or 3 weeks later than those of `Kokei No. 14' from which it originated. Morphological characteristics of the canopy and leaf photosynthetic rates of `KM95-A68' were similar to those of `Kokei No. 14'. No apparent differences were observed in the anatomy of root cross sections of `KM95-A68' and `Kokei No. 14'. An apparent increase in the root zeatin riboside (ZR) levels were observed in `Kokei No. 14' at storage root formation. Root ZR levels differed between `Kokei No. 14' and `KM95-A68'. The onset of increase in root ZR levels was delayed by 2 or 3 weeks in `KM95-A68' in comparison to `Kokei No. 14'. Maximum root ZR levels in `Kokei No. 14' were 2.2 times higher in comparison to `KM95-A68'. This appeared to be a factor in delayed storage root formation of `KM95-A68'. Results of reciprocal grafts of `KM95-A68' and `Kokei No. 14' indicated that the late storage root-forming characteristic of `KM95-A68' is a characteristic that arises from the root itself.
Grzegorz Bartoszewski, Cesar V. Mujer, Katarzyna Niemirowicz-Szczytt, and Ann C. Smigocki
A Lycopersicon esculentum Mill. (tomato) cDNA clone with high similarity to a Nicotiana plumbaginifolia Viv. (tobacco) cytochrome P450 gene was isolated using 5' and 3' rapid amplification of cDNA ends (RACE). The isolated cDNA (GenBank Accession No. AF249329) has an open reading frame of 1494 base pairs (bp) and encodes a protein of 498 amino acids with 75% identity to the N. plumbaginifolia cytochrome P450 (CYP72A2) and 45% to a Catharanthus roseus G. Don (Madagaskar periwinkle) CYP72A1 protein sequence. By Southern-blot analysis, one or two highly homologous genes were detected in the L. esculentum genome. Expression of the cloned P450 gene was regulated by circadian rhythm and enhanced by wounding. Leaf transcripts were detected in the light but not dark. Highest transcript levels were observed 3 hours after mechanical wounding. No increase in expression was seen in response to applications of zeatin as with the N. plumbaginifolia gene. Of the tissues analyzed, shoot tips and young leaves and fruit had the highest detectable transcript levels. Attempts to transform more than 1400 cotyledon explants of L. esculentum with sense or antisense CYP72A2 gene constructs produced no transgenic plants.
Mari Tahara, Takeshi Yasuda, Naotsugu Uchida, and Tadashi Yamaguchi
Somatic embryos were regenerated from protoplasts isolated from embryogenic callus on young leaf explants from mature coffee trees. Embryos were regenerated on modified Murashige and Skoog medium supplemented with 5 μm BA. Somatic embryos developed into intact plants. Mannitol at 0.5 m was adequate as an osmoticum for isolating protoplasts, but subsequent culture required 0.3 m mannitol. A culture system in which osmolality was decreased gradually accelerated formation of colonies and somatic embryogenesis. Chemical name used: N-(phenylmethyl)-1H-purine-6-amine (BA).
Michael A. Arnold and Eric Young
CuCO3 at 100 g·liter-1 in a paint carrier applied to interior container surfaces effectively prevented root deformation in container-grown Malus domestica Borkh. and Fraxinus pennsylvanica Marsh. seedlings. CuCO3 treatments nearly doubled the number of white unsuberized root tips in both species. CuCO3 treatment increased some measures of root and shoot growth before and after transplanting to larger untreated containers. Root pruning at transplanting tended to reduce root and shoot fresh and dry matter accumulation in F. pennsylvanica seedlings and shoot extension in M. domestica seedlings. In some cases, root pruning of M. domestics at transplanting from CuCO3-treated containers increased root growth compared to unpruned CuCO3-treated and untreated seedlings. Changes in growth induced by CuCO3 and root pruning were not related to changes in trans -zeatin riboside-like activity in the xylem sap of-apple.
Don C. Elfving and Dwayne B. Visser
The height above the bud union at which induced feathers develop on fruit trees in the nursery is an important determinant of tree quality for an intended market. The bioregulators cyclanilide (CYC; Bayer Environmental Science, Research Triangle Park, NC) and a proprietary formulation of 6-benzyladenine and gibberellins A4 and A7 (Promalin [PR]; Valent BioSciences, Walnut Creek, CA) affected the final height above the union of the lowest induced sylleptic shoot (feather) differently in apple and sweet cherry trees in the nursery. In apple, both products resulted in the lowest induced feather developing at approximately 4 to 20 cm below the height of the central leader shoot tip at the time of bioregulator application. In sweet cherry, the lowest induced feather typically originated starting approximately 2 to 20 cm above the central leader shoot tip height at the time of bioregulator application. Nursery tree height can serve as a suitable criterion for timing bioregulator applications to obtain feathers starting within a specific range of height above the bud union as long as species-specific feathering response characteristics are taken into account. Chemical names used: 1-(2,4-dichlorophenylaminocarbonyl)-cyclopropane carboxylic acid (Cyclanilide), N-(phenylmethyl)-1H-purine-6-amine + gibberellins A4A7 (Promalin), polyoxyethylenepolypropoxypropanol, dihydroxypropane, 2-butoxyethanol (Regulaid).
Yasuyoshi Hayata, Yoshiyuki Niimi, and Naoto Iwasaki
Applying a 200 ppm solution of CPPU to pollinated ovaries of watermelon (Citrullus lunatus Matsum) at anthesis increased fruit set from 26.9% (control) to 95%. Applying CPPU solutions to nonpollinated ovaries at anthesis induced parthenocarpy, yielding 65% and 89.5% fruit set, respectively with 20 and 200 ppm applications. However, 64% of the 20 ppm CPPU-treated parthenocarpic fruit stopped growth 10 days after treatment. Growth of CPPU-treated, pollinated, and nonpollinated fruit increased significantly compared with growth of control fruit during the first 10 days after treatment, but, except for the 20 ppm CPPU parthenocarpic fruit, growth subsequently slowed, resulting in fruit equal in size to the control by harvest. CPPU application did not affect soluble solids content of pollinated fruit, but reduced content of parthenocarpic fruit treated with 20 ppm. Fructose content was generally higher than glucose and sucrose at harvest. However, in pollinated fruit treated with 20 ppm CPPU, sucrose levels were higher than glucose and fructose. These results suggest that CPPU is practical for promoting fruit set and seedless fruit without adversely affecting fruit quality and development.
John Erwin, Rene O’Connell, and Ken Altman
Photoperiod, irradiance, cool temperature (5 °C), and benzyladenine (BA) application effects on Echinopsis ‘Rose Quartz’ flowering were examined. Plants were placed in a 5 °C greenhouse under natural daylight (DL) for 0, 4, 8, or 12 weeks, then moved to a 22/18 °C (day/night temperature) greenhouse under short days (SD, 8-hour DL) plus 0, 25, 45, or 75 μmol·m−2·s−1 supplemental lighting (0800–1600 hr; 8-hour photoperiod), long days (LD) delivered with DL plus night-interruption lighting (NI) (2200–0200 hr), or DL plus 25, 45, or 75 μmol·m−2·s−1 supplemental lighting (0800–0200 hr) for 6 weeks. Plants were then grown under DL only. Percent flowering plants increased as irradiance increased from 0–25 to +75 μmol·m−2·s−1 on uncooled plants, from 0% to 100% as 5 °C exposure increased from 0 to 8 weeks under subsequent SD and from 25% to 100% as 5 °C exposure increased from 0 to 4 weeks under subsequent LD. As 5 °C exposure duration increased from 0 to 12 weeks (SD-grown) and from 0 to 8 weeks (LD-grown), flower number increased from 0 to 11 and from 5 to 21 flowers per plant across irradiance treatments, respectively. Total production time ranged from 123 to 147 days on plants cooled from 8 to 12 weeks (SD-grown) and from 52 to 94 days on plants cooled for 0–4 weeks to 119–153 days on plants cooled for 8–12 weeks (LD-grown). Flower life varied from 1 to 3 days. BA spray application (10–40 mg·L−1) once or twice after a 12-week 5 °C exposure reduced flower number. Flower development was not photoperiodic. High flower number (17–21 flowers/plant) and short production time (including cooling time, 120–122 days) occurred when plants were grown at 5 °C for 8 weeks, then grown under LD + 45–75 μmol·m−2·s−1 for 6 weeks (16 hours; 10.9–12.8 mol·m−2·d−1) at a 22/18 °C day/night temperature. Taken together, Echinopsis ‘Rose Quartz’ exhibited a facultative cool temperature and facultative LD requirement for flowering.
Thomas J. Zabadal and Martin J. Bukovac
The effects of CPPU [forchlorfenuron, N-(2-chloro-4-pyridinyl)-N-phenylurea] on berry development of Vitis labrusca and V. labrusca × V. vinifera cultivars was evaluated under field conditions. A concentration response was initially established by spraying clusters of `Himrod' at a mean berry diameter of about 5 mm with 0, 5, 10, or 15 mg·L–1 CPPU. Berry enlargement was monitored (16, 30, 44, and 59 days after treatment) during development. Cluster mass, number of berries per cluster, berry mass and firmness, and °Brix were determined at harvest. Berry mass was dramatically increased (2.3 versus about 3.6 g/berry) at harvest by all concentrations of CPPU. Cluster mass and compactness were also increased and berry firmness was linearly related to CPPU concentration (r 2 = 0.997). There was no significant effect on number of berries per cluster (79 to 86). °Brix, rachis necrosis at harvest, and berry abscission after 30 days of refrigerated storage (1 °C) were significantly reduced. Effect of time of CPPU application (0, 5, and 10 mg·L–1) was established by treatment of clusters at mean berry diameters of about 4, 5, 7, and 9 mm. Response was indexed by following berry enlargement at 14, 28, 42, and 56 (maturity) days after treatment. Maximum berry size for both 5 and 10 mg·L–1 was obtained from applications at 4 to 7 mm berry diameter. Relative response of seedless and seeded cultivars was compared by application of CPPU at 0, 5, 10, or 15 mg·L–1 to clusters (4 to 6 mm berry diameter) of seedless `Vanessa' and `Lakemont' and seeded `Concord' and `Niagara'. Bioresponse was determined by a time course of berry enlargement and berry and cluster mass, number of berries per cluster, and rating cluster compactness at maturity. Except for `Lakemont' at the 5 mg·L–1 concentration, CPPU at all concentrations increased seedless berry diameter significantly from the first measurement at 14 through 56 days after application. Berry and cluster mass and cluster compactness were significantly increased in `Vanessa'. In contrast, the only effect of CPPU on the two seeded cultivars was an increase in berry size in `Concord' and an initial increase in berry size 14 days after application in `Niagara'.