Generation of chemiluminescence during enzymatic isolation of protoplasts from leaves of Oryza sativa

Butt, Adrian D.; Bestwick, Charles

doi:10.1016/s0176-1617(97)80290-8

Cited by 6 publications

(3 citation statements)

References 30 publications

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“…The isolation procedure can also induce further chromatin condensation and the apoptotic pathway [72]. This condensation is accompanied by a reduction in the scavenging capacity of reactive oxygen species (ROS) [73], which leads to an increased ROS accumulation [74,75]. Recently, the presence of chromatin condensation has also been demonstrated during protoplast isolation followed by subsequent cultivation in a buffer without PGRs [76].…”

Section: The Protoplast Isolation Step As the Key For Reprogrammingmentioning

confidence: 99%

From Single Cell to Plants: Mesophyll Protoplasts as a Versatile System for Investigating Plant Cell Reprogramming

Pasternak

Lystvan

Betekhtin

et al. 2020

IJMS

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Plants are sessile organisms that have a remarkable developmental plasticity, which ensures their optimal adaptation to environmental stresses. Plant cell totipotency is an extreme example of such plasticity, whereby somatic cells have the potential to form plants via direct shoot organogenesis or somatic embryogenesis in response to various exogenous and/or endogenous signals. Protoplasts provide one of the most suitable systems for investigating molecular mechanisms of totipotency, because they are effectively single cell populations. In this review, we consider the current state of knowledge of the mechanisms that induce cell proliferation from individual, differentiated somatic plant cells. We highlight initial explant metabolic status, ploidy level and isolation procedure as determinants of successful cell reprogramming. We also discuss the importance of auxin signalling and its interaction with stress-regulated pathways in governing cell cycle induction and further stages of plant cell totipotency.

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Section: The Protoplast Isolation Step As the Key For Reprogrammingmentioning

confidence: 99%

From Single Cell to Plants: Mesophyll Protoplasts as a Versatile System for Investigating Plant Cell Reprogramming

Pasternak

Lystvan

Betekhtin

et al. 2020

IJMS

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“…45 Also, high levels of antioxidant response machinery in protoplasts were discovered in multiple species. [46][47][48][49] To these studies, we add that the protoplast cytosol is likely acidified, affecting the chloroplast membrane lipid composition, and having the potential to interfere with a wide swathe of protein functions.…”

Section: Protoplast Cytosolic Ph Is Decreasedmentioning

confidence: 99%

An Arabidopsis protoplast isolation method reduces cytosolic acidification and activation of the chloroplast stress sensor SENSITIVE TO FREEZING 2

Barnes

Elowsky

Roston

2019

Plant Signaling & Behavior

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Chloroplasts adapt to freezing and other abiotic stresses in part by modifying their membranes. One key-remodeling enzyme is SENSITIVE TO FREEZING2 (SFR2). SFR2 is unusual because it does not respond to initial cold stress or cold acclimation, instead it responds during freezing conditions in Arabidopsis. This response has been shown to be sensitive to cytosolic acidification. The unique lipid products of SFR2 have also been detected in response to non-freezing stresses, but what causes SFR2 to respond in these stresses is unknown. Here, we investigate protoplast isolation as a representative of wounding stress. We show that SFR2 oligogalactolipid products accumulate during protoplast isolation. Notably, we show that protoplast cytosol is acidified during isolation. Modification of the buffers reduces oligogalactolipid accumulation, while prolonged incubation in the isolated state increases it. We conclude that SFR2 activation during protoplast isolation correlates with cytosolic acidification, implying that all SFR2 activation may be dependent on cytosolic acidification. We also conclude that protoplasts can be more gently isolated, reducing their stress.

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“…Previous study revealed that ROS were generated during enzymatic isolation of oat mesophyll protoplasts [ 40 ]. Thereafter ROS accumulation was also reported in isolated protoplasts of other plants, such as grapevine, tobacco, rapeseed, and rice [ 41 – 43 ]. Furthermore, increased activity of antioxidant machinery was observed in isolated protoplasts of most plants [ 40 , 41 , 44 , 45 ].…”

Section: Introductionmentioning

confidence: 99%

Identification of protoplast-isolation responsive microRNAs in Citrus reticulata Blanco by high-throughput sequencing

Zhou

et al. 2017

PLoS ONE

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Protoplast isolation is a stress-inducing process, during which a variety of physiological and molecular alterations take place. Such stress response affects the expression of totipotency of cultured protoplasts. MicroRNAs (miRNAs) play important roles in plant growth, development and stress responses. However, the underlying mechanism of miRNAs involved in the protoplast totipotency remains unclear. In this study, high-throughput sequencing technology was used to sequence two populations of small RNA from calli and callus-derived protoplasts in Citrus reticulata Blanco. A total of 67 known miRNAs from 35 families and 277 novel miRNAs were identified. Among these miRNAs, 18 known miRNAs and 64 novel miRNAs were identified by differentially expressed miRNAs (DEMs) analysis. The expression patterns of the eight DEMs were verified by qRT-PCR. Target prediction showed most targets of the miRNAs were transcription factors. The expression levels of half targets showed a negative correlation to those of the miRNAs. Furthermore, the physiological analysis showed high levels of antioxidant activities in isolated protoplasts. In short, our results indicated that miRNAs may play important roles in protoplast-isolation response.

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Generation of chemiluminescence during enzymatic isolation of protoplasts from leaves of Oryza sativa

Cited by 6 publications

References 30 publications

From Single Cell to Plants: Mesophyll Protoplasts as a Versatile System for Investigating Plant Cell Reprogramming

From Single Cell to Plants: Mesophyll Protoplasts as a Versatile System for Investigating Plant Cell Reprogramming

An Arabidopsis protoplast isolation method reduces cytosolic acidification and activation of the chloroplast stress sensor SENSITIVE TO FREEZING 2

Identification of protoplast-isolation responsive microRNAs in Citrus reticulata Blanco by high-throughput sequencing

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