Summary
When cotton (Gossypium hirsutum, DPL 90ne) ovules are removed from the carpel on the day of anthesis and placed on an appropriate medium, fiber cells will differentiatein vitro from the ovule epidermis. Alpha-amanitin, an inhibitor of poly (A)+ RNA synthesis was added to ovule cultures at selected times after culture initiation to determine the timing of events in fiber development. Replicate cultures were initiated from day of anthesis, cotton ovules with α-amanitin being added to media at final concentrations ranging from 0.05μg/ml to 5.0 μg/ml. After 21 d in culture, ovules were evaluated for fiber length, percentage of ovules producing fiber, fiber dry weight, and fiber cellulose content. Cultures treated with α-amanitin on the day of anthesis to 2 d postanthesis were prevented from forming normal numbers of fiber cells. Addition of α-amanitin to cultures between 2 and 10 d postanthesis reduced the length that fiber cells attained in culture after 21 d. Addition of α-amanitin to cultures at 4 d postanthesis had as much an effect on the accumulation of cellulose in the fiber cell wall as the later addition of the inhibitor at 8 to 10 d postanthesis. The inhibitory effect of α-amanitin on fiber growth was reversible when ovules were transferred to amanitin-free medium within 2 d after culture initiation. Ovules placed on media containing α-amanitin for 5 d or longer could not overcome the inhibitory effects when transferred to media lacking the inhibitor. These results indicate critical time periods when RNA transcripts important for specific phases of development are being synthesized.
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Beasley, C. A.; Ting, I. P. Effects of plant growth substances onin vitro fiber development from unfertilized cotton ovules. Am. J. Bot. 60:188–194; 1974.
Beasley, C. A.; Ting, I. P.; Linkins, A. E., et al. Cotton ovule culture: a review of progress and a preview of potential. In: Street, H. E., ed. Tissue culture and plant science. New York: Academic Press; 1974:169–192.
Berlin, J. D. The outer epidermis of the cottonseed. In: Mauney, J. R.; Stewart, J. M., eds. Cotton physiology. The Cotton Foundation Reference Book Series. Memphis, TN: The Cotton Foundation; 1986:375–414.
Cheung, C. P.; Wu, J.; Suhadolnik, R. J. Dependence of protein synthesis on RNA synthesis during early hours of germination of wheat embryos. Nature 277:66–67; 1979.
Cochet-Meilhac, M.; Chambon, P. Animal DNA-dependent RNA polymerases: 11. Mechanism of the inhibition of RNA polymerases B by amatoxins. Biochim. Biophys. Acta 353:160–184; 1974.
Dhindsa, R. S. Hormonal regulation of cotton ovule and fiber growth: effects of bromodeoxyuridine, AMO-1618 andp-chlorophenoxyisobutyric acid. Planta 141:269–272; 1978.
Dhindsa, R. S.; Beasley, C. A.; Ting, I. P. Effects of abscisic acid on in vitro growth of cotton fiber. Planta 130:197–201; 1976.
Graves, D.; Stewart, J. M. Chronology of the differentiation of cotton (Gossypium hirsutum L.) fiber cells. Planta 175:254–258; 1988.
Jendrisak, J. The use of α-amanitin to inhibitin vivo RNA synthesis and germination in wheat embryos. J. Biol. Chem. 255:8529–8533; 1980.
Koizuka, N.; Tanaka, Y.; Morohashi, Y. Expression of alpha-amylase in response to wounding in mung bean. Planta 195:530–534; 1995.
Lohmann, J.; Schickle, H.; Bosch, R. REN display, a rapid and efficient method for nonradioactive differential display and mRNA isolation. BioTechniques 18:200–202; 1995.
Meinert, M.; Delmer, D. Changes in the biochemical composition of the cell wall of the cotton fiber during development. Plant Physiol. 59:1088–1097; 1977.
Pitto, L.; Lo Schiavo, F.; Giuliano, G., et al. Analysis of the heat-shock protein pattern during somatic embryogenesis of carrot. Plant Mol. Biol. 2:231–237; 1983.
Schubert, A. M.; Benedict, C. R.; Berlin, J. D., et al. Cotton fiber developmental kinetics of cell elongation and secondary wall thickening. Crop Sci. 13:704–709; 1973.
Spiegel, S.; Marcus, A. Polyribosome formation in early wheat embryo germination independent of either transcription or polyadenylation. Nature 256:228–230; 1975.
Triplett, B. A.; Timpa, J. Characterization of cell-wall polymers from cotton ovule culture fiber cells by gel permeation chromatography. In Vitro Cell. Dev. Biol. 31P:171–175; 1995.
Updegraff, D. M. Semi-micro determination of cellulose in biological materials. Anal. Biochem. 32:420–424; 1969.
Weiland, T.; Faulstich, H. Amatoxins, phallotoxins, phallolysin, and antamanide: the biologically active components of poisonousAmanita mushrooms. Crit. Rev. Biochem. 5:185–260; 1978.
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Triplett, B.A. Stage-specific inhibition of cotton fiber development by adding α-amanitin to ovule cultures. In Vitro Cell.Dev.Biol.-Plant 34, 27–33 (1998). https://doi.org/10.1007/BF02823119
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DOI: https://doi.org/10.1007/BF02823119