Establishment and characterization of Pinus pinaster suspension cell cultures

Azevedo, Herlânder; Dias, Alberto Carlos Pires; Tavares, R. M.

doi:10.1007/s11240-008-9349-1

Cited by 16 publications

(9 citation statements)

References 24 publications

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“…The calculated t d for the cell suspension culture was 6.6 days and their l was 0.104 days -1 . A similar behavior was previously reported in the establishment of other cell suspension cultures (Azevedo et al 2008;Estrada-Zuñiga et al 2009). On the other hand, the cell viability remained around 70% throughout the 21 days of culture.…”

Section: Callus Inductionsupporting

confidence: 86%

Establishment and characterization of Prosopis laevigata (Humb. & Bonpl. ex Willd) M.C. Johnst. cell suspension culture: a biotechnology approach for mesquite gum production

et al. 2011

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This study presents a protocol for the establishment of Prosopis laevigata cell suspension culture as a strategy to obtain an in vitro mesquite gum productive cell line. The callus used for this purpose was obtained with hypocotyls from 15-day-old plantlets, placed on Murashige-Skoog medium with two different plant growth regulators (PGRs), 2,4,5-trichlorophenoxy acetic acid (2,4,5-T; 5.0 lM) and kinetin (KIN; 5.0 lM). With this PGRs treatment, after four subcultures (30 days each) an exuded gum-like substance was observed on the callus surface. The growth kinetics of the cell suspension culture showed a specific cell growth rate (l) of 0.14 d -1 and doubling time (t d ) of 6.6 days, respectively. The gum-like substance from callus culture and the broth from cell suspension culture were subjected to chemical analysis and compared with the mesquite gum exuded from wild trees. Both, gum-like substance from callus culture and the broth from cell suspension culture showed the presence of Arabinogalactanproteins, and their polysaccharide fraction presented the same monosaccharides as those isolated from mesquite gum. In addition, the emulsifying properties of gum-like substance from callus culture and the broth from cell suspension culture were compared to those of mesquite gum and all three samples exhibited similar emulsifying capacity and emulsification stability.

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Section: Callus Inductionsupporting

confidence: 86%

Establishment and characterization of Prosopis laevigata (Humb. & Bonpl. ex Willd) M.C. Johnst. cell suspension culture: a biotechnology approach for mesquite gum production

et al. 2011

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“…Carbon starvation in cultures of marine pine ( Pinus pinaster Ait.) was suggested to induce PCD events (Azevedo et al, 2008; Azevedo et al, 2014).…”

Section: Discussionmentioning

confidence: 99%

Vacuolar processing enzyme translocates to the vacuole through the autophagy pathway to induce programmed cell death

Teper‐Bamnolker

Danieli

Peled-Zehavi

et al. 2019

Preprint

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One sentence summary: Carbon starvation induced programmed cell death 26 by trafficking vacuolar processing enzyme through the autophagy pathway to 27 the vacuole. 28 29The author responsible for distribution of materials integral to the findings 30 presented in this article in accordance with the policy described in the 31 Instructions for Authors (www.plantcell.org) is: Dani Eshel (dani@agri.gov.il). 32 33 34 2 35 Abstract 36 37 The caspase-like vacuolar processing enzyme (VPE) is a key factor in 38 programmed cell death (PCD) associated with plant stress responses. Growth 39 medium lacking a carbon source and dark conditions caused punctate labeling 40 of 35S::VPE1-GFP (StVPE1-GFP) in potato leaves. Carbon starvation of BY-2 41 cells induced higher VPE activity and PCD symptoms. Growing VPE-RNAi BY-42 2 cells without sucrose reduced VPE activity and prevented PCD symptoms. 43 During extended exposure to carbon starvation, VPE expression and activity 44 levels peaked, with a gradual increase in BY-2 cell death. Histological analysis 45 of StVPE1-GFP in BY-2 cells showed that carbon starvation induces its 46 translocation from the endoplasmic reticulum to the central vacuole, through 47 tonoplast engulfment. Exposure of BY-2 culture to the autophagy inhibitor 48 concanamycin A caused autophagic bodies accumulation in the cell vacuole. 49 Such accumulation did not occur in the presence of 3-methyladenine, an 50 inhibitor of early-stage autophagy. BY-2 cells constitutively expressing 51 StATG8IL-RFP, an autophagosome marker, showed colocalization with the 52 StVPE1-GFP protein in the cytoplasm and vacuole. RNAi silencing of the core 53 autophagy component ATG4 in BY-2 cells reduced VPE activity and cell death. 54 These results are the first to suggest that VPE translocates to the cell vacuole 55 through the autophagy pathway, leading to PCD. 56 endopeptidase activity, unlike caspases that cleave their substrates at aspartic 91 acid residues (Silva et al., 2005; Van Durme and Nowack, 2016). The VPE 92 proteins belong to a family of cysteine proteinases that are well conserved 93 among a variety of organisms, including many plant and animal species (Cai 94 and Gallois, 2015; Hatsugai et al., 2015; Sueldo and van der Hoorn, 2017). 95 VPEs were originally found to be responsible for the maturation of seed 96 storage proteins and various other vacuolar proteins in plants (Hara-Nishimura 97 et al., 1991; Hara-Nishimura et al., 1993; Hatsugai et al., 2004). VPE, which is 98 released into the vacuole during PCD, triggers the degradation of other 99 proteins (Hara-Nishimura et al., 2005; Kuroyanagi et al., 2005; Van Durme and 100 Nowack, 2016). VPEs, which exhibit caspase-1-like activity, play important 101 roles in plant PCD, be it developmental or in response to biotic or abiotic stress 102 (reviewed by Hatsugai et al., 2015; Vorster et al., 2019). Specifically, VPE has 103 been characterized as a major factor in the HR. By silencing the gene 104 encoding VPE, Hatsugai et al. (2004) showed that vacuolar collapse, caused ...

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“…Pinus pinaster heterotrophic suspension cells have been successfully used to study plant-pathogen interactions and changes in nutrient uptake profiles (Azevedo et al, 2008a;2008b). In the present study, this model was used to analyse the homeostasis of ROS during salt stress under nonphotosynthetic conditions.…”

Section: Discussionmentioning

confidence: 99%

“…A heterotrophic Pinus pinaster suspension cell culture was previously established in our laboratory from root segments of maritime pine seedlings (Azevedo et al, 2008a). Suspension cells were maintained in Murashige and Skoog (MS) medium (Murashige and Skoog, 1962) supplemented with 3% glucose, 5 mg L −1 dithiothreitol, 0.1 g L −1 myo-inositol, 1 mg L −1 BA and 2 mg L −1 2,4-D, at pH 6.…”

Section: P Pinaster Suspension Culture Maintenance and Characterizationmentioning

confidence: 99%

Effect of salt on ROS homeostasis, lipid peroxidation and antioxidant mechanisms in Pinus pinaster suspension cells

2009

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Abstract• In the Pinus genus, information on the effectiveness of oxidative defence mechanisms during exposure to salt is lacking. The effect of salt stress imposition on ROS homeostasis was investigated using maritime pine (Pinus pinaster Ait.) suspension cells as a model system.• Cells were maintained in MS-based medium, exposed to salt (50, 100 and 150 mM NaCl) and analysed for biomass production, evidencing a decreasing growth capacity. Use of 100 mM NaCl imposed severe salt stress without affecting cell viability, being chosen for subsequent studies on the ROS homeostasis of salt shock-treated suspension cells.• Increased total ROS levels were evident on the second day of salt exposure, but a superoxide ion transient burst was immediately noticeable. Additionally, lipid peroxide formation seemed to correlate with superoxide ion breakdown. In-gel superoxide dismutase activity evidenced a FeSOD homodimer with strongly increasing activity between hours 12-48 of salt stress imposition. Subsequently, P. pinaster Fe-Sod1 and csApx1 genes were isolated from a cDNA library and expression was shown to increase within 12-24 h.• Results show that severe salt treatment generates oxidative stress in P. pinaster cells despite the induction of antioxidant systems, and suggest a putative involvement of ROS in salt stress signalling. Mots-clés :H 2 O 2 / pin maritime / NaCl / superoxide Résumé -Effet du stress salin sur l'homéostasie des formes réactives d'oxygène, la peroxydation des lipides et les mécanismes antioxydants dans des suspensions cellulaires de Pinus pinaster.• Les informations sur les mécanismes de défense oxydative du pin en réponse à un stress salin sont rares. L'effet d'une exposition au sel sur l'homéostasie des formes réactives d'oxygène (FRO) a été étudié en utilisant une suspension cellulaire de pin maritime (Pinus pinaster Ait.) comme modèle.

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Establishment and characterization of Pinus pinaster suspension cell cultures

Cited by 16 publications

References 24 publications

Establishment and characterization of Prosopis laevigata (Humb. & Bonpl. ex Willd) M.C. Johnst. cell suspension culture: a biotechnology approach for mesquite gum production

Establishment and characterization of Prosopis laevigata (Humb. & Bonpl. ex Willd) M.C. Johnst. cell suspension culture: a biotechnology approach for mesquite gum production

Vacuolar processing enzyme translocates to the vacuole through the autophagy pathway to induce programmed cell death

Effect of salt on ROS homeostasis, lipid peroxidation and antioxidant mechanisms in Pinus pinaster suspension cells

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