Conditions for Strict Autotrophic Culture of Tobacco Callus

McHale, Neil A.

doi:10.1104/pp.77.1.240

Cited by 13 publications

(6 citation statements)

References 6 publications

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“…In small Acer pseudoplatanus cultures, a sharp peak in growth occurs at CO 2 concentrations as low as 1 % (v/v), others (Table 54). Photoautotrophic Chenopodium rubrum cultures also correspond to such cells in their membrane potential which exceeds -200 m V, their hexose transport supported by carriers and dependent on protons, and their content of cell wall-bound acidic invertases (Hiisemann and Barz 1977;McHale 1985;Rogers et al 1988).…”

Section: Carbon Sourcementioning

confidence: 93%

Plant Cell Biotechnology

Endreß

1994

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Section: Carbon Sourcementioning

confidence: 93%

Plant Cell Biotechnology

Endreß

1994

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“…They were transferred to polyurethane pads in Petri plates (15) containing liquid medium without sucrose and were grown in air for 1 week in light until they were green. Small (5 mm) plantlets were screened for 02-resistant growth by transferring them to illuminated (100 umol photons m-2 s-') Plexiglas chambers that were flushed continuously for 2 weeks with moistened 42% 02 containing sufficicnt CO2 to maintain the concentration in the chamber at about 160 ,uL/L at 290C.…”

Section: Growth Of Plantsmentioning

confidence: 99%

Selection and Characterization of Tobacco Plants with Novel O₂-Resistant Photosynthesis

Zelitch

1989

Plant Physiol.

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Plants were obtained with novel O2-resistant photosynthetic characteristics. At low CO2 (250-350 MiL CO2 L-1) and 300C when 02 was increased from 1% to 21% to 42%, the ratio of net CO2 uptake in 02-resistant whole plants or leaf discs compared to wild type increased progressively, and this was not related to stomatal opening. Dihaploid plantlets regenerated from anther culture were initially screened and selected for 02-resistant growth in 42% 02/ 160 ML CO2 L-1 and 0.18% of the plantlets showed 02-resistant photosynthesis. About 30% of the progeny (6 of 19 plants) of the first selfing of a fertile plant derived from a resistant dihaploid plant had 02-resistant photosynthesis, and after a second selfing this increased to 50% (6 of 12 plants). In 21% 02 and low C02, net photosynthesis of the resistant plants was about 15% greater on a leaf area basis than wild type. Net photosynthesis was compared in leaf discs at 30 and 380C in 21% 02, and at the higher temperature 02-resistant plants showed still greater photosynthesis than wild type. The results suggest that the O2-resistant photosynthesis described here is associated with a decreased stoichiometry of CO2 release under conditions of rapid photorespiration. This view was supported by the finding that leaves of 02-resistant plants averaged 40% greater catalase activity than wild type.Photosynthesis in C3 plants is inhibited 33 to 55% by the 21% 02 in the atmosphere compared with rates at 1 to 3% 02. To call attention to the importance of this effect, I have referred to it as oxygen stress (28). Inhibition by 21% 02 is completely reversed by high C02, showing that it is related to photorespiration. One obvious way of decreasing this adverse effect of 02 in normal CO2 would be to increase the C02/02 specificity of ribulose bisphosphate carboxylase/oxygenase (10).In the oxidative photorespiratory pathway, two glycine molecules condense to produce one seine and one C02; hence, the stoichiometry should be 25% of the carbon flux if this is the sole source of photorespiratory CO2 (26). However, recent experiments conducted at low CO2 concentrations (5, 6) indicate that the stoichiometry of photorespiration in vivo can exceed 25% when the ratio of net photorespiration/ net photosythesis is elevated by high 02 (above 21%) or higher temperatures. This suggests an alternative approach to the improvement of photosynthetic efficiency in C3 plants based on identification of mutants that avoid this increase in stoichiometry of CO2 release under conditions of rapid photorespiration.Here it is demonstrated that atmospheres of 42% 02/low CO2 inhibit growth of tobacco seedlings, and that the inhibition is reversed by high CO2. I have therefore screened a population of dihaploid tobacco plantlets for survival and growth in a 42% 02/low CO2 atmosphere and studied the characteristics of selected plants with 02-resistant photosynthesis in populations of two generations of fertile plants derived from a dihaploid. These plants exhibit superior net photosynthesis under certain co...

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“…Callus cultures were initiated on a Linsmaier and Skoog (LS) medium (7) from leaf sections of a dihaploid plant obtained from anther cultures (6) of Nicotiana tabacum, L. (cv Havana Seed). Subsequent establishment of autotrophic cultures on polyurethane pads was as previously described (8). Autotrophic callus was grown (liquid LS medium; 0.3 mg-L-' NAA,' no sucrose) in plexiglas chambers (31 L) flushed (100 ml.…”

Section: Methodsmentioning

confidence: 99%

Effects of CO₂ and O₂ on Photosynthesis and Growth of Autotrophic Tobacco Callus

McHale¹,

Zelitch²,

Peterson³

1987

Plant Physiol.

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Autotrophic culture of plant cells in vitro represents a potentially valuable system for genetic manipulation ofphotosynthesis. This system is particularly amenable to alteration of photosynthetic functions governed by chloroplast-encoded enzymes, as seen in the recent recovery of maternally inherited triazine resistant mutants from photomixotrophic cultures oftobacco (3).We have continued to investigate photorespiratory characteristics ofautotrophic tobacco callus as a step toward development of systems for in vitro selection of mutants with alterations in photorespiratory metabolism. Previous demonstrations of inhibition of "'CO2 uptake by 100% 02, light-dependent 02 stimulation of "'CO2 release into C02-free air, and glycolate accumulation after treatment with a a-hydroxy-2-pyridinemethanesulfonate (1) provided evidence for the existence ofphotorespiration in autotrophic callus. In addition, it was shown that 60% 02 completely inhibited autotrophic growth in tobacco callus, but had no effect on heterotrophic growth.To investigate the suitability of high 02 as a selective agent for identification of photorespiratory mutants, we have examined the effects of various 02 levels on growth and net photosynthesis in autotrophic tobacco callus, with particular attention to CO2 reversal of 02 effects. We show that inhibition of net photosynthesis by 40% 02 is fully reversible by high CO2. In contrast, the inhibition ofautotrophic growth by 40% 02 is not fully reversible by high C02, indicating that a portion of this 02 toxicity arises from a mechanism other than photorespiration. Results showing no 02 inhibition of heterotrophic growth, however, confirmed previous observations that 02 toxicity is mediated through photosynthesis, perhaps through Mehler-type reactions in the chloroplasts. MATERIALS AND METHODSCulture Method. Callus cultures were initiated on a Linsmaier and Skoog (LS) medium (7) from leaf sections of a dihaploid plant obtained from anther cultures (6) of Nicotiana tabacum, L. (cv Havana Seed). Subsequent establishment of autotrophic cultures on polyurethane pads was as previously described (8). Autotrophic callus was grown (liquid LS medium; 0.3 mg-L-' NAA,' no sucrose) in plexiglas chambers (31 L) flushed (100 ml. min-') with 1% C02/21% 02 under continuous illumination (150 E.m 2. s') for 2 weeks prior to measurements of net photosynthesis. Growth experiments were conducted in chambers flushed with the specified atmospheres, and data were collected on total fresh weight and sample dry weight at the end of three weeks.Gas Exchange Analysis. Measurements of net photosynthesis and dark respiration were performed on autotrophic callus at 30°C in one liter Plexiglas chambers fitted with stopcocks, a fan, and a rubber serum stopper. The chamber was flushed (1 L min-') with moistened gas of known composition, and measurements were begun by closing the stopcocks and removing gas samples at intervals ofseveral minutes through the serum stopper with a hypodermic syringe. Assimilation of CO2 was determined...

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Conditions for Strict Autotrophic Culture of Tobacco Callus

Cited by 13 publications

References 6 publications

Plant Cell Biotechnology

Plant Cell Biotechnology

Selection and Characterization of Tobacco Plants with Novel O₂-Resistant Photosynthesis

Effects of CO₂ and O₂ on Photosynthesis and Growth of Autotrophic Tobacco Callus

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Conditions for Strict Autotrophic Culture of Tobacco Callus

Cited by 13 publications

References 6 publications

Plant Cell Biotechnology

Plant Cell Biotechnology

Selection and Characterization of Tobacco Plants with Novel O2-Resistant Photosynthesis

Effects of CO2 and O2 on Photosynthesis and Growth of Autotrophic Tobacco Callus

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Product

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Selection and Characterization of Tobacco Plants with Novel O₂-Resistant Photosynthesis

Effects of CO₂ and O₂ on Photosynthesis and Growth of Autotrophic Tobacco Callus