Regeneration of plants from cultured guard cell protoplasts of Nicotiana glauca (Graham)

Sahgal, Prateek; Martinez, Gregory V.; Roberts, C H; Tallman, Gary

doi:10.1016/0168-9452(94)90057-4

Cited by 21 publications

(21 citation statements)

References 23 publications

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“…Vol. 109, 1995, the viability of cultured cells (Cupples et al, 1991), and the survival of cells in culture (Sahgal et al, 1994). Among the more important determinants of survival was the temperature at which GCP were cultured (this study).…”

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confidence: 81%

“…GCP are totipotent (Sahgal et al, 1994;Hall et al, 1995), suggesting that a subset of the genome is used to maintain guard cells in their uniquely differentiated state. Neither genes that differentiate guard cells from other plant cells nor the mechanisms resulting from the activity of those genes have been identified.…”

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confidence: 99%

“…As culture temperatures were increased from 24 to 32"C, survival for 1 week in culture increased from approximately 20% to approximately 80% of the cells used to initiate the culture. At a11 of those temperatures, approximately 90% of the cells surviving underwent cytokinininduced cell division to form the type of callus tissue that could be used to regenerate plants (Sahgal et al, 1994). GCP cultured at 32°C increased nearly 30-fold in "footprint area" (hereafter called "cell growth").…”

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confidence: 99%

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Temperature and Abscisic Acid Can Be Used to Regulate Survival, Growth, and Differentiation of Cultured Guard Cell Protoplasts of Tree Tobacco

et al. 1995

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Cuard cell protoplasts isolated from leaves of Nicofiana g h c a (Craham) were cultured. Conditions were sought that would maximize survival and maintain cells i n their differentiated state. Temperature was an important determinant of survival, growth, and differentiation. As temperatures were increased from 24 to 32'C, survival for 1 week in culture increased from approximately 20% to approximately 80% of cells used to initiate cultures. At all of these temperatures, approximately 90% of surviving cells divided to form callus tissue. "Footprint" areas of cells cultured for 1 week at 32°C increased almost 30-fold. Cells cultured for 1 week at 34 t o 40°C also survived i n high percentages (approximately 80%), but they retained a morphology similar to that of guard cells and they did not divide. Footprint areas of cells cultured for 1 week at 38°C increased 6-fold. Cells cultured at 36 t o 4OoC in media containing 0.1 or 1.0 W M abscisic acid survived in high percentages and did not divide. At 38°C their footprint areas did not increase, but cells so cultured increased in diameter when treated with fusicoccin. Morphologies and electrophoretic profiles of total sodium dodecyl sulfate-extractable proteins suggest that cells cultured at 38°C in media containing abscisic acid remain differentiated. ~-a-(2-Aminoethoxyviny1)-glycine reduced survival t o <1 O/O at 26 or 32°C but had no effect at 38OC. At lower temperatures, cell growth and survival appear to be ethylene dependent.

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confidence: 81%

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confidence: 99%

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confidence: 99%

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Temperature and Abscisic Acid Can Be Used to Regulate Survival, Growth, and Differentiation of Cultured Guard Cell Protoplasts of Tree Tobacco

et al. 1995

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“…Totipotency of protoplasts from guard cells of tobacco has also recently been reported (Sahgal et al, 1994). However, from both of these papers it is unclear as to what potential role the physical process of protoplast isolation played in this phenomenon.…”

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confidence: 99%

Stomatal Guard Cells Are Totipotent

et al. 1996

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It has been successfully demonstrated, using epidermis explants of sugar beet (Befa vulgaris L.), that stomatal guard cells retain full totipotent capacity. Despite having one of the highest degrees of morphological adaptation and a unique physiological specialization, it is possible to induce a re-expression of full (embryogenic) genetic potential in these cells in situ by reversing their highly differentiated nature to produce regenerated plants via a callus stage. The importance of these findings both to stomatal research and to our understanding of cytodifferentiation in plants is discussed.

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“…At 32°C, in a medium containing an auxin (1-naphthaleneacetic acid [NAA]) and a cytokinin (6-benzylaminopurine [BAP]), GCP survive in high percentages (70%-80%) for up to 1 month (Roberts et al, 1995). Under the influence of NAA and BAP, GCP expand 20-to 30-fold, regenerate cell walls, dedifferentiate, and divide to form callus from which plants can be regenerated (Sahgal et al, 1994). Both NAA and BAP must be supplied exogenously before GCP will survive and undergo the extensive cellular remodeling that accompanies dedifferentiation and division (Gushwa et al, 2003).…”

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Heat Suppresses Activation of an Auxin-Responsive Promoter in Cultured Guard Cell Protoplasts of Tree Tobacco

et al. 2007

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Cultured guard cell protoplasts (GCP) of tree tobacco (Nicotiana glauca) comprise a novel system for investigating the cell signaling mechanisms that lead to acquired thermotolerance and thermoinhibition. At 32°C in a medium containing an auxin (1-naphthaleneacetic acid [NAA]) and a cytokinin (6-benzylaminopurine), GCP expand, regenerate cell walls, dedifferentiate, and divide. At 38°C, GCP acquire thermotolerance within 24 h, but their expansion is limited and they neither regenerate walls nor reenter the cell cycle. These putative indicators of auxin insensitivity led us to hypothesize that heat suppresses induction of auxin-regulated genes in GCP. Protoplasts were transformed with BA-mgfp5-ER, in which the BA auxin-responsive promoter regulates transcription of mgfp5-ER encoding thermostable green fluorescent protein (GFP) or with a similar 35S-cauliflower mosaic virus constitutive promoter construct. Heat suppressed NAA-mediated activation of BA. After 21 h at 32°C in media with NAA, 49.0% 6 3.9% of BA-mgfp5-ER transformants strongly expressed GFP; expression percentages were similar to those of 35S-mgfp5-ER transformants at 32°C or 38°C. After 21 h at 38°C in media with NAA, 7.9% 6 1.6% of BA-mgfp5-ER transformants weakly expressed GFP, similar to GCP cultured at 32°C in media lacking NAA. Expression at 38°C was not increased by incubating for 48 h or increasing NAA concentrations 20-fold. After 9 to 12 h at 38°C, BA was no longer activated when cells were transferred to 32°C. Heat-stressed cells accumulate reactive oxygen species, and hydrogen peroxide (H 2 O 2 ) suppresses auxin-responsive promoter activation in Arabidopsis (Arabidopsis thaliana) mesophyll protoplasts. H 2 O 2 did not suppress BA activation at 32°C, nor did superoxide and H 2 O 2 scavengers prevent BA suppression at 38°C.

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Regeneration of plants from cultured guard cell protoplasts of Nicotiana glauca (Graham)

Cited by 21 publications

References 23 publications

Temperature and Abscisic Acid Can Be Used to Regulate Survival, Growth, and Differentiation of Cultured Guard Cell Protoplasts of Tree Tobacco

Temperature and Abscisic Acid Can Be Used to Regulate Survival, Growth, and Differentiation of Cultured Guard Cell Protoplasts of Tree Tobacco

Stomatal Guard Cells Are Totipotent

Heat Suppresses Activation of an Auxin-Responsive Promoter in Cultured Guard Cell Protoplasts of Tree Tobacco

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