Five greenhouse and two Geld experiments were conducted to evaluate tissue culture-propagated (TC) raspberry (Rubus idaeus cv. Heritage) sensitivity to preemergent herbicides. Plant performance was measured by plant vigor, above-ground fresh weight, root development, and primocane number. Simazine and oryzalin caused significant injury to newly planted TC raspberry plants in greenhouse and field experiments. The severity of injury was generally linear with respect to herbicide rate, but no appreciable differences in injury were observed between the granular and spray applications. Napropamide wettable powder caused some foliar injury, but plants recovered within one growing season and growth was equal or superior to the hand-weeded controls. The granular formulation of napropamide produced similar results, but did not cause the initial foliar burn. Pre-plant dipping of roots into a slurry of activated carbon did not prevent simazine or oryzalin injury, but injury was reduced when herbicide applications were delayed. Simazine applied 4 weeks after planting was not Injurious, and oqzalin applied 2 or 4 weeks after planting caused some foliar injury, hut no reduction in plant fresh weight. Delayed treatments of napropamide increased foliar injury. Herbicide tolerance of tissue-cultured plantlets appeared to be less than that of conventionally propagated plants. Chemical names used: N,N-diethyl-2-(1-napthalenyloxy)propanamide (napropamide), 4-(dipropylamino)-3,5-dinitrobenzenesulfonamide (oryzalin), 6-chloro-N,N'diethyl-1,3,5-triazine-2,4-diamine (simazine).
Joseph C. Neal, Marvin P. Pritts, and Andrew F. Senesac
Sitheswary Logendra, Mei-Mann Hsueh, and Harry W. Janes
The effect of root mass on tomato fruit size in tissue culture was studied. The root mass of the ovaries was changed either by growing in culture media containing different concentrations of NAA (α– napthaleneacetic acid) or by culturing the ovaries with and without sepals. The root mass increased with a decrease in NAA concentration from 10.0 to 2.5 μM and the ovaries with sepals developed more roots. The tomato fruit size was affected by the root mass. The greater the root mass, the larger was the fruit size. However, the larger fruit size from ovaries cultured with sepals could be attributed either to the presence of more roots (greater absorption of sucrose) or to the sepal (additional carbon fixation by photosynthesis), or to both the sepals and more roots. Moreover, it is possible that the presence of sepals induce root development. These results indicate that the presence of sepals and total root mass are two important factors that influence the fruit size in vitro.
Nguyen Phuc Huy, Vu Quoc Luan, Le Kim Cuong, Nguyen Ba Nam, Hoang Thanh Tung, Vu Thi Hien, Dung Tien Le, Kee Yoeup Paek, and Duong Tan Nhut
-elongated ex vitro explants as the source under dark–light cycles for plant regeneration through internode tissue cultures. The results of this study provide a new approach to micropropagation of P. callosum for commercial propagation. Materials and Methods
Christopher J. Currey and Nicholas J. Flax
(photoperiod) or cool temperatures (vernalization) for flower induction and development. The new ‘Ladyslippers’ streptocarpus series are propagated using tissue culture ( Uhl, 2012 ). While these plants produce showy finished flowering plants, they regularly
Dae-Geun Oh, Maria Chrzastek, and Edward C. Tigchelaar
Heterozygous multiple marker genetic stocks were synthesized by crossing three multiple genetic marker stocks to a common inbred parent PU812. The four parents and 3 F1's were cultured to obtain regenerants from leaf discs. Fifty four regenerants were derived from 3 F1's and 12 from the 4 parents, Among the regenerants, 16 plants were identified as tetraploid (24.2%); low fertility was usually associated with tetraploidy, however there were a few exceptions.
Selfed seeds, identified by cluster number, were harvested from sexual F1's and R0 plants for F2 progeny tests for the known marker genes. While there were no abnormal segregations for marker genes in the sexual progenies, 13 of 46 progenies from tissue culture derived regenerants showed significant deviations from expected normal segregations for a number of markers. Two of the abnormal progenies were identified as tetraploids by root-tip examinations; segregation ratios fit duplex random chromatid segregation for gene a on chromosome 11 and random chromosome segregation for gene c on chromosome 6. The cams of abnormal segregations in other progenies remain unknown. Results suggest that unknown genetic events arising during tissue culture may distort segregations for marker genes in the subsequent sexual progeny of tissue culture regenerants.
In Pelargonium, the plastid mutation in three independent cell layers L1, L2, and L3, can produce plastid chimeras with visible shoot colour difference such as GWG (green-white-green) and GGW (green-green-white). Chimera can be used to trace the relationship between the cell layers of different genotypes during shoot development and the effect of the mutated genes on shoot development. In this study, we have obtained different adventitious shoots with GGG, GWG, GGW, and WWW combinations of cell layers through tissue culture of petioles and internodes from GGW and GWG chimeras of Pelargonium zonale `Mrs Pollock'. Much higher percentage (14.9%) of chimeral adventitious shoots was obtained from GGW tissues than from GWG tissues (4.2%). Of the 10.8% chimeral adventitious shoots regenerated in this experiment, 8.6% are different from the original type of explants. This result indicated that cells at least in both L2 and L3 of the explants were involved in the regeneration of the adventitious shoots. The number of shoot types regenerated is likely dependent on the number and the type of cells that were in direct contact with the culture medium. It is suggested that the mixed cells can be used to produce the chimera by tissue culture. Three possible ways to form the chimeras in vitro culture were discussed. Chemical names used: TDZ =1-phenyl-3-(1,2,3-thiadiazol-5-yl)urea (Thidiazuron); IAA = Indole-3-acetic acid; PVP = polyvinylpyrrolidone.
Hazel Y. Wetzstein, Choongsik Kim, and Harry E. Sommer
Effects of autoclaving volume, gelling agent (Bactoagar, Gel-gro, Phytagar), and basal salts [Murashige and Skoog (MS); Woody Plant Medium (WPM); Gamborg B5 (GB)] on gel strength and pH of tissue culture media were tested. Gel strength was significantly affected by gelling agent and basal medium. MS media were generally softer than comparable WPM or GB media. As the vessel volume during autoclaving decreased, gel strength significantly decreased with Phytagar and Bactoagar gelling agents; Gel-gro had greater gel strength at the intermediate volume of medium autoclave. In all cases, autoclaving resulted in a pH decrease of 0.2 to 0.5 pH units. Lower pH values were associated with softer gels. The type of gelling agent did not greatly affect the postautoclave pH; mean values among gelling agents were within 0.05 pH units. Postautoclave pH of MS medium was lower than that of WPM or GB. This study verifies the need to observe uniform sterilization protocols to maintain consistency in the chemical and physical properties of media.
Sitheswary Logendra, Mei-Mann Hsueh, and Harry W. Janes
Growing tomato fruits in tissue culture, using ovaries, could be used as a model system to study fruit development and sink strength/activity. Producing a “normal and healthy” fruit is essential in developing this system. Many factors affect the growth and development of the fruit. The objective of this study was to investigate the effect of the age of the ovary (i.e., the number of days after pollination) on growth and final fruit size. The results indicate that the fruit size, root development, and uniformity in growth of the fruit were affected by the initial age of the ovary. The older the ovary, the greater was the final fruit size and uniformity. The development of root mass was not affected by the age of the ovary until 7 days of pollination. However, root development was suppressed in ovaries that were of 9 days after pollination. The fruits from younger ovaries were more irregular in shape. All the fruits from ovaries harvested at 9 days after pollination were more uniform and round as compared to other treatments.
Ahmed El-Shiekh, David K. Wildung, James J. Luby, Kay L. Sargent, and Paul E. Read
Plants of `Northblue' blueberry, propagated in tissue culture (TC) or from softwood, single-node cuttings (ST), were evaluated in field plantings established in 1984 at Becker and Grand Rapids, in central and northern Minnesota, respectively. Plantings were observed from 1987 through 1994 to determine the persistence of such effects as increased vigor, more spreading growth habit, and higher yield observed for TC plants during the initial 3 years after planting. TC plants had significantly higher yields at Grand Rapids in 1989 and 1994. At Grand Rapids, the consistently greater plant spread (bearing area) of TC plants resulted in higher yields of TC plants over all years combined. At Becker, TC and ST plants did not differ for plant height or spread after 10 years and, in 2 of 5 years, ST plants had heavier average berry weights. At Grand Rapids, TC plants did not differ consistently in height, or subjective ratings of the amount of bloom or crop. The effects of propagation method on yield and growth habit of `Northblue' are limited to early years in warmer locations, but can be of longer-term significance in colder areas with shorter growing seasons and lower winter temperatures, where plant spread is a more important factor than plant height in determining yield.
F.A. Hammerschlag and R. Scorza
Abbreviations: TC, tissue culture; TCSA, trunk cross-sectional area. Mention of a trademark, proprietary product, or vendor by the U.S. Dept. of Agriculture does not imply its approval to the exclusion of other products or vendors that may also be