Oxidative stress could be responsible for the recalcitrance of plant protoplasts

Papadakis, Anastasia K.; Roubelakis‐Angelakis, Kalliopi A.

doi:10.1016/s0981-9428(02)01423-7

Cited by 66 publications

(40 citation statements)

References 77 publications

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“…Cultivars with high starch content are known to be less amenable to protoplast isolation, and it is known that no starch or few starchy cells usually yield much higher protoplast numbers (Crowder et al, 1979). The process of protoplast isolation per se is a highly stress-inducing procedure, which may cause more damage in cells with high content of starch (Papadakis et al, 2002). 'Valencia' was the most recalcitrant genotype for protoplast isolation and it was excluded from this study.…”

Section: Resultsmentioning

confidence: 99%

Citrus asymmetric somatic hybrids produced via fusion of gamma-irradiated and iodoacetamide-treated protoplasts

Bona

Gould

Miller

et al. 2009

Pesq. agropec. bras.

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-The objective of this study was to produce citrus somatic asymmetric hybrids by fusing gamma-irradiated protoplasts with iodoacetamide-treated protoplasts. Protoplasts were isolated from embryogenic suspension cells of grapefruit (Citrus paradisi Macfad.) cultivars Ruby Red and Flame, sweet oranges (C. sinensis Osbeck) 'Itaboraí', 'Natal', Valencia', and 'Succari', from 'Satsuma' (C. unshiu Marcow.) and 'Changsha' mandarin (C. reticulata Blanco) and 'Murcott' tangor (C. reticulata x C. sinensis). Donor protoplasts were exposed to gamma rays and receptor protoplasts were treated with 3 mmol L -1 iodoacetamide (IOA), and then they were fused for asymmetric hybridization. Asymmetric embryos were germinated, and the resulting shoots were either grafted onto sour orange, rough lemon or 'Swingle' (C. paradisi x Poncirus trifoliata) x 'Sunki' mandarin rootstock seedlings, or rooted after dipping their bases in indol-butyric acid (IBA) solution. The products were later acclimatized to greenhouse conditions. Ploidy was analyzed by flow cytometry, and hybridity was confirmed by amplified fragment length polymorphism (AFLP) analysis of plantlet DNA samples. The best treatment was the donor-recipient fusion combination of 80 Gy-irradiated 'Ruby Red' protoplasts with 20 min IOA-treated 'Succari' protoplasts. Tetraploid and aneuploid plants were produced. Rooting recalcitrance was solved by dipping shoots' stems in 3,000 mg L -1 IBA solution for 10 min.

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Section: Resultsmentioning

confidence: 99%

Citrus asymmetric somatic hybrids produced via fusion of gamma-irradiated and iodoacetamide-treated protoplasts

Bona

Gould

Miller

et al. 2009

Pesq. agropec. bras.

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“…We also kept in mind that if transcription is still taking place, which is possible for many transcripts, then our estimated stress half-lives represent an underestimation. A contributing factor for this stark difference may be that it is difficult to accurately measure the half-life of lowly expressed transcripts in the absence of stress in a transcriptional shutoff experiment (Narsai et al, 2007), and transcripts could have altered stability in cell culture (Papadakis and Roubelakis-Angelakis, 2002;Xu et al, 2013). Nevertheless, while the most unstable mRNA in cell culture has a half-life of 13.0 min, we identified 71 transcripts with an estimated half-life NL of lower than 13.0 min and 222 transcripts with a half-life NL of under 60.0 min.…”

Section: Rapid Excess-light Recovery Is Underpinned By Highly Unstablmentioning

confidence: 99%

Rapid Recovery Gene Downregulation during Excess-Light Stress and Recovery in Arabidopsis

et al. 2017

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Stress recovery may prove to be a promising approach to increase plant performance and, theoretically, mRNA instability may facilitate faster recovery. Transcriptome (RNA-seq, qPCR, sRNA-seq, and PARE) and methylome profiling during repeated excess-light stress and recovery was performed at intervals as short as 3 min. We demonstrate that 87% of the stress-upregulated mRNAs analyzed exhibit very rapid recovery. For instance, HSP101 abundance declined 2-fold every 5.1 min. We term this phenomenon rapid recovery gene downregulation (RRGD), whereby mRNA abundance rapidly decreases promoting transcriptome resetting. Decay constants (k) were modeled using two strategies, linear and nonlinear least squares regressions, with the latter accounting for both transcription and degradation. This revealed extremely short half-lives ranging from 2.7 to 60.0 min for 222 genes. Ribosome footprinting using degradome data demonstrated RRGD loci undergo cotranslational decay and identified changes in the ribosome stalling index during stress and recovery. However, small RNAs and 5ʹ-3ʹ RNA decay were not essential for recovery of the transcripts examined, nor were any of the six excess light-associated methylome changes. We observed recovery-specific gene expression networks upon return to favorable conditions and six transcriptional memory types. In summary, rapid transcriptome resetting is reported in the context of active recovery and cellular memory.

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“…Moreover, POD activity was found to be significantly higher in dividing protoplasts than in non-dividing (Xu et al, 2013). POD activity could also be considered a predictor of hydrogen peroxide accumulation during isolation and culture (Papadakis and Roubelakis-Angelakis, 2002). In this respect, the accumulation of ROS could be related to reprogramming of the developmental pathway of the cell (Jones and Smirnoff, 2005).…”

Section: Discussionmentioning

confidence: 99%

“…Reactive oxygen species (ROS) play a crucial role in plant morphogenesis and development, acting as signaling molecules at the level of both the plant cell and the whole organism (Jones and Smirnoff, 2005), and therefore may be involved in determining the regeneration ability of the protoplasts. Studies on non-regenerating and regenerating protoplasts revealed that increasing accumulation of toxic oxygen during isolation is likely to cause protoplast recalcitrance (Sinimis et al, 1994;Papadakis et al, 2002;Xu et al, 2013). It has also been shown in protoplasts of barley that a failure of an antioxidative defense mechanism is associated low cell viability and mitotic arrest (Kapur et al, 1993).…”

Section: Introductionmentioning

confidence: 99%

Activity of selected components of antioxidant system in grass pea and yellow lupine protoplasts after enzymatic isolation

Wiszniewska¹,

Piwowarczyk²

2015

bta

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In this study, the performance of the antioxidant system in protoplasts of grass pea (Lathyrus sativus L.) and yellow lupine (Lupinus luteus L.) isolated using solutions of different compositions was evaluated. Protoplasts were isolated from leaves of grass pea and cotyledons of yellow lupine seedlings grown in vitro. Hydrolytic enzymes were dissolved in three different solutions (ultrapure water, inorganic salt solution (CPW), and inorganic salt solution with organic additives (C)). Protoplasts purified using these solutions were subjected to the analyses of peroxidase (POD) and radical scavenging activities. Phenolic profile was also determined. In grass pea, the highest POD was determined in protoplasts isolated in CPW salt solution, while in yellow lupine, the other two solutions were found superior. Protoplasts isolated in CPW salt solution had the highest content of phenolic compounds, ranging from 28.8 to 40.1 and from 32.0 to 58.0 mg/g fresh weight (f.w.) in lupine and grass pea, respectively. Moreover, in protoplasts obtained using CPW solution, the total antioxidant activity was the highest in all tested genotypes as expressed by high proportion of scavenged 2,2,-diphenyl-1-picrylhydrazyl (DPPH) radical. The results showed that changes in composition of the isolation solutions affected the level and activity of selected components belonging to cell antioxidant system in legume protoplasts. Considering the recalcitrance of two studied plants in protoplast culture, this finding is an important piece of information because events occurring at the stage of isolation may affect further development of protoplasts in the culture.

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Oxidative stress could be responsible for the recalcitrance of plant protoplasts

Cited by 66 publications

References 77 publications

Citrus asymmetric somatic hybrids produced via fusion of gamma-irradiated and iodoacetamide-treated protoplasts

Citrus asymmetric somatic hybrids produced via fusion of gamma-irradiated and iodoacetamide-treated protoplasts

Rapid Recovery Gene Downregulation during Excess-Light Stress and Recovery in Arabidopsis

Activity of selected components of antioxidant system in grass pea and yellow lupine protoplasts after enzymatic isolation

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