Susceptibility of tomato (Lycopersicon esculentum Mill) genotpyes to the root-knot nematode Meloydogyne incognita and to heat stress can be evaluated in a single labor- and time-saving operation using a nondestructive in vitro excised root technique. Seeds are sterilized and germinated for 2 days on 1% water agar. Five-mm root sections are grown at 28 and 35 C for 30 days on Gamborg-B medium with and without nematode inoculum. Evaluation criteria include fresh and dry weight and the appearance of juveniles, adults, gulls, and egg masses. Evidence will be presented on the breakdown of resistance to M. incognita under high temperature stress.
Aref A. Abdul-Baki, S. A. Haroon, and R. N. Huettel
Sarbagh Salih, Howard Waterworth, and Daniel A. Thompson
Mingbo Qin, Chiwon W. Lee, Alex Y. Borovkov, and Murray E. Duysen
A study was initiated to characterize key enzymes that influence sweetness in carrot (Daucus carota L.) roots. Sucrose synthase (SS), sucrose phosphate synthase (SPS), and UDP-glucose pyrophosphorylase (UDPL) genes were isolated from potato (Solanum tuberosum L.) and cloned in an anti-sense orientation into Agrobacterium tumefaciens Bin19, which has a CaMV 35S promoter. Seedling hypocotyl sections of selected carrot lines were pre-incubated on B5 medium for 2 days, co-cultivated with A. tumefaciens Bin 19 for additional 3 days, and then transferred to a modified B5 medium containing 50 g/mL kanamycin and 400 g/mL carbenicillin. In 4 weeks, 18.6%, 33.3%, and 26.7% of the cultures from a breeding line (W204-C) were found to be transformed, respectively, with SS, SPS, and UDPL as determined by kanamycin resistance. In contrast, no kanamycin-resistant calli were obtained from a commercial cultivar (Navajo) in these transformation studies. The transformed calli proliferated in the medium containing 50 g/mL kanamycin and 400 g/mL carbenicillin, whereas non-transformed calli died in the same medium. These transformed calli are currently being used to regenerate plants via asexual embryogenesis using a suspension culture. The influence of these additional genes on sugar metabolism and accumulation in root tissues of transformed carrots will be characterized in the future.
Dirk R. Vuylsteke and Rodomiro Ortiz
In vitro-propagated plants of plantain (Musa spp., AAB group) did not manifest consistently superior horticultural performance compared to conventional propagules. Tissue culture plants grew vigorously and taller than sucker-propagated plants, but higher yield was not obtained, probably because of severe disease and suboptimal husbandry input. Phenotypic variation was higher in tissue culture plants, although this increase was not always statistically significant. There were no other detrimental effects of in vitro propagation on field performance. Botanical seed set rates for the two types of propagules were similar. The advantages of tissue-culture-derived plants as improved planting material would be most relevant for establishing field nurseries for further clean, conventional propagation of newly bred or selected genotypes.
Joe-Ann McCoy and N.D. Camper
Hypericum perforatum L. (St. John's Wort) has an extensive history as an important medicinal herb used for the treatment of neurological and depressive disorders (Linde et al., 1996). The objective of this study was to establish an in vitro tissue culture protocol for St. John's Wort. Nodal segments, axillary buds, and leaf disc explants produced multiple shoots and callus on Murashige and Skoog minimal organics medium supplemented with combinations of indoleacetic acid (IAA; 0.57, 2.85, 5.71 μm) and benzylaminopurine (BA; 2.22, 4.44, 8.88 μm). Shoot production occurred on all combinations of IAA/BA tested and was significantly less in treatments without hormones. Callus production was higher on treatments containing 2.85 μm IAA + 4.44 μm BA, or 5.71 μm IAA + 8.88 μm BA. Shoots transferred to hormone-free medium at 8 weeks formed roots by 12 weeks. A micropropagation protocol was established for St. John's Wort using mature plants as the explant source.
Marianela Ramirez, Marek J. Krasowski, and Judy A. Loo
Heuser, 1987 ). The purpose of this study was to further the development of vegetative propagation techniques for disease-free American beech, including tissue culture and grafting. This was done by: Experimenting with techniques to decrease in
Mohd Faisal, Naseem Ahmad, and Mohammad Anis
1 To whom reprint requests should be addressed; e-mail email@example.com . The authors thank A K. Sharma, Deputy Director and Head, Tissue Culture Laboratory, National Botanical Research Institute (CSIR
Xabier Barandiaran, Nieves Martín, María Fernanda Rodríguez-Conde, Antonio Di Pietro, and Jesus Martín
The influence of different callus induction media on the regeneration process in garlic was tested. The auxin 2,4 dichlorophenoxyacetic acid frequently used in garlic tissue culture was found to be detrimental when used at the levels described in the literature. However, combinations of growth regulators commonly used for dicot tissue culture produced high levels of callus induction and regeneration that could be used efficiently in a transformation program.
Ravindra K. Hajela, Neerja Hajela, Mark G. Bolyard, Wayne M. Barnes, and Mariam B. Sticklen
A simple gene transfer method based on Agrobacterium -mediated transformation of adventitious multiplication of Juneberry (Amelanchier laevis L.) basal shoots is described. Evidence is presented for successful integration and expression of a transformed gene in greenhouse-grown transgenic plants. This method can transform woody perennials that are difficult to regenerate from leaf disks, protoplasts, or other tissue culture regimens.