Welcome
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110/09/13
Man,Man,
Happy ManHappy Man
210/09/13
Plant!!?Plant!!?
310/09/13
KITTUR RANI CHANNAMMA COLLEGE OF
HORTICULTURE, ARABHAVI
Programmed Cell Death in PlantsProgrammed Cell Death in Plants
410...
TOPIC DIVISION
510/09/13
Definition
Programmed cell death (PCD) is a physiological cell death process
involved in the selective elimination of unwa...
10/09/13 7
History of Programmed cell death / apoptosis
1800s - Numerous observation of cell death
1842 - Carl Vogt, first time descr...
Death by Injury vs. Death by Suicide
(Necrosis vs. Apoptosis)
(Kerr et al., 1992, Germany) 910/09/13
Fig -1 : Difference b...
Necrosis vs. ApoptosisNecrosis vs. Apoptosis
• Cellular condensation
• Membranes remain intact
• Requires ATP
• Cell is ph...
10/09/13 11
http://science.howstuffworks.com/life/cellular-microscopic/apoptosis.htm
Fig -2 : Phases of PCD in a Vascular ...
Why should a cell decide to commit suicide?Why should a cell decide to commit suicide?
PCD is needed for proper developmen...
PCD Regulators
Caspase
(Cysteinyl aspartate-specific
proteases)
Bcl-2 family Signal transduction
pathway
Caspase-8
Caspase...
10/09/13 14
Programmed cell death
Programmed cell death : Pathways
Death
Ligands
Effector
Caspase 3
Death
Receptors
Initiator
Caspase 8
PCDPCD
Mitochondria/...
Fig-3 : Activation of apoptosis from outside the cell.
1610/09/13
APOPTOSIS: important in embryogenesis
Morphogenesis (eliminates excess cells):
Selection (eliminates non-functional cells)...
APOPTOSIS: important in embryogenesis
Immunity (eliminates dangerous cells):
Self antigen
recognizing cell
Organ size (eli...
APOPTOSIS: important in adults
Tissue remodeling (eliminates cells no longer needed):
Virgin mammary gland Late pregnancy,...
Fig -4 : Sites of PCD in a Vascular Plant.
The orange spheres represent internal dead cells, and the branched structures
o...
Fig -5 : Functions of PCD in Plants. The red regions represent cells that have been targeted
for PCD and the orange region...
Aleuron layer
In seeds , aleuron cells form a secretary tissue
that releases hydrolases to digest the endosperm
and nouris...
Root Cap CellsRoot Cap Cells
Cell death occurs in root caps when roots are grown,Cell death occurs in root caps when roots...
(A) Electron micrograph of a living cell.
(B) Electron micrograph of a dead cell. In the dead cell, the plasma membrane (p...
Senescence
 Senescence is the final phase of plant vegetative
and reproductive development, preceding the widespread
deat...
10/09/13 26
Vascular plants transport water in columns of specialized dead cells termed TEs.
Differentiation of TEs involv...
INTERACTIONS WlTH THE ENVIRONMENT
Hypoxia (O2 deprivation)
 Cell death can occur in the cortex of the root and stem base ...
10/09/13 28
Plant-Pathogen interactions
Plants can recognize certain pathogens and activate defenses
(called the resistanc...
Case studies
2910/09/13
Tomato: Lycopersicon esculantum cv. Zhonghusihao
Grown in GH 260
C / 180
C , day/ night and natural day length and stored ...
Fig-7: Measurement of relative ion leakage from pericarp discs of tomato fruit as
an indicator of cell death by measuring ...
Fig-8: In-situ detection of DNA fragmentation by TUNEL analysis in
tomato fruit. (Gui et al., 2009, U.S.A)
a–c Pericarp sa...
Fig-9 : In-situ detection of DNA fragmentation by TUNEL analysis in tomato fruit.
(Gui et al., 2009, U.S.A)10/09/13
Caspase-1 (Ac-YVAD-AMC), caspase-3 (Ac-DEVD-AMC), caspase-8 (Ac-IETD-
AMC)
and caspase-9 (Ac-LEHD-AMC) , HS - Treated at 4...
3510/09/13
Fig -11 : Effect of Z-VAD-FMK pretreatment on cell death in heat treated tomato
pericarp discs. . (Gui et al., ...
Materials and methods:
Tomato ( Lycopersicon esculentum Mill.) cell suspension cultures, line Msk 8
(Koornneef et al., 198...
Fig -12 : Induction of cell death in tomato suspension cells by Fluorescine
diacetate (FDA )staining method.
□ - Staurospo...
STAU- Staurosporin-2µM
CPT - Camptothesin-5µM
FUM - Fumonisin-20µM
FeSo4 -100mM
STAU- Staurosporin-2µM
CPT - Camptothesin-...
5µM -Camptothecin (CPT),
50µM- N-ethylmaleimide (NEM),
5µM -iodoacetamide (IA),
200µM- Nm Nα-p-tosyl- L-lysine
chloromethy...
CPT - Camptothesin-5µM
(CPT+ YVAD) – 100nM Caspase
specific peptide inhibitor- Acyl -Tyr-
Val-Ala-Asp- chloromethylketone ...
4110/09/13
Tomato seeds were incubated in the dark @ 250
c prior to use. Isolation of
endosperm tissue was performed asept...
Fig -16 : Post-germinative tomato endosperm cells display features of reserve mobilization
and PCD including SlCysEP-conta...
Conti….
d)Localization of SlCysEP within ricinosomes is
evidenced by the white arrowheads lying within
the ricinosomes,
e)...
Fig-18 : SlCysEp transcript and enzyme accumulation in tomato seeds.
(Christoper et al.,2013, Heidelberg)
Fig-18 : SlCysEp...
Fig -19 : Tissue specific SlCysEP expression and enzyme accumulation using
Northern-blot analysis
(Christoper et al.,2013,...
a) Non-germinated (NG), b) short seedling (SS), c) long seedling (LS)
Endosperm halves from NG (d), SS , LS (f) seeds. (*)...
4710/09/13
Fig-21 :Quantification of protien and SlCysEP profile isolated on en equal tissue basis.
(Christoper et al.,201...
Tomato (Lycopersicon esculentum Mill.) cell suspension culture,
line Msk8, was grown in a liquid MS medium + vitamins and ...
Fig -22: Effect of cadmium salts on cell death in tomato suspension cells 24 h after
the treatments.
(Iakimova et al., 200...
Fig -23: Morphological features of living (A) and dead (B) tomato cells 24 h after
treatment with 100 μM CdSO4.
( Iakimova...
Fig-24: Kinetic of hydrogen peroxide production in tomato suspension cells
exposed to 100 μM CdSO4. ( Iakimova et al., 200...
Fig -25: Kinetic of ethylene production in tomato suspension cells exposed
to 100 μM CdSO4.
( Iakimova et al., 2009, Bulga...
Objectives : Elicitors of different origin (fumonisin B1, fungal toxin), camptothecin
(alkaloid from Camptotheca acuminata...
Fig-26 : Effect of 20 μM Fumonisin B1 (FumB1), 5 μM camptothecin (CPT),
100 μM CdSO4 or 5 μM mastoparan (MP) on cell death...
Fig -27 : Effect of 100 nM Ac-YVAD-CMK, 100 nM Z-Asp-CH2-DCB and 1mM (AEBSF) on
cell death in tomato suspension cells indu...
5610/09/13
 PCD is essential for ensuring the proper
development of plant
 Interaction with environment which results ro...
10/09/13 57
GERMAN SCIENTIST CARL VOGT WAS FIRST TO
DESCRIBE THE PRINCIPLE OF APOPTOSIS IN 1842.
GERMAN SCIENTIST CARL VOG...
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  • WELCOME
  • Prospects and Retrospectes of oil palm cultivation in India
  • (Ellis et al., 1991).
  • Is it having long history
  • (Kerr et al., 1992, Germany)
  • Jean et al., 1996, USA
  • Xylogenesis
  • Case studies
  • Tomato: Lycopersicon esculantum cv. Zhonghusihao Grown in GH 26 0 C / 18 0 C , day/ night and natural day length and stored @ 25 0 C . Devided in to groups (5 in each). In the heat shock treatment, fruit bag sub merged in water bath for 20 min @ 45 0 C, the control was placed @ 25 0 C for 20 min.
  • Tomato ( Lycopersicon esculentum Mill. ) cell suspention cultures, line Msk 8 (Koornneef et al 1987 Murashige and Skoog liquid medium Cells were used for experiments 5 d after sub culture. Cell death inducers and inhibiters were added simultaniously to 5 ml of suspension culture. 5% percent buffer formalin for detection of nuclear changes and DNA fragmentation
  • Figure: Induction of cell death in tomato suspension cells by FDA staining method. □ - ∆ - Anke et al., 2000, Netherland X - O -
  • 5 µM camptothesin (CPT), 50 µM N-ethylmaleimide (NEM), 5 µM iodoacetamide (IA), 200 µM Nm N α -p-tosyl- L-lysine chloromethylekitone (TLCK), 1mM 4,2,amiloethyle benzensulfonyl fluoride (AEBSF) 20 µM fumonisin BI (FUM) 2 µM staurosporine (STAU)
  • Materials
  • Conti….
  • Iakimova et al., 2009, Bulgaria
  • Conti….
  • Programmed cell death in plants by shivanand b. koppad

    1. 1. Welcome ²æà ¹zÉÝñÀég À 110/09/13
    2. 2. Man,Man, Happy ManHappy Man 210/09/13
    3. 3. Plant!!?Plant!!? 310/09/13
    4. 4. KITTUR RANI CHANNAMMA COLLEGE OF HORTICULTURE, ARABHAVI Programmed Cell Death in PlantsProgrammed Cell Death in Plants 410/09/13
    5. 5. TOPIC DIVISION 510/09/13
    6. 6. Definition Programmed cell death (PCD) is a physiological cell death process involved in the selective elimination of unwanted cells. Ellis et al., (1991) Necrosis is a non physiological process involving cell swelling, lysis, and the inflammatory leakage of cell contents. Cohen, (1993) Programmed Cell DeathProgrammed Cell Death (PCD ……….?)Programmed Cell DeathProgrammed Cell Death (PCD ……….?) Programmed cell death (PCD)/ apoptosis Programmed cell death occurs via apoptosis - a Greek word meaning “falling leaves” - John et al., (1972)
    7. 7. 10/09/13 7
    8. 8. History of Programmed cell death / apoptosis 1800s - Numerous observation of cell death 1842 - Carl Vogt, first time describe the principles of apoptosis. 1885 - Anatomist Walther Flemming delivered a more precise description of the process of programmed cell death. 1908 - Mechnikov wins Nobel prize (phagocytosis). 1930-40 - Studies of metamorphosis. 1948-49 - Cell death in chick limb. 1955 - Beginning of studies of Ricinosomes. 1964-66 - Necrosis & PCD described. 1971 - Term apoptosis coined in Greece. 1977 - Cell death genes in C. elegans. 1980-82 - DNA ladder observed & ced-3 identified. 1989-91 - Apoptosis genes identified, including bcl-2, fas/apo1 & p53, ced-3 sequenced. 2002 - Jone E Sultan, won the nobel prize in medicine for his poineering research810/09/13
    9. 9. Death by Injury vs. Death by Suicide (Necrosis vs. Apoptosis) (Kerr et al., 1992, Germany) 910/09/13 Fig -1 : Difference between necrosis and PCDFig -1 : Difference between necrosis and PCD
    10. 10. Necrosis vs. ApoptosisNecrosis vs. Apoptosis • Cellular condensation • Membranes remain intact • Requires ATP • Cell is phagocytosed, no tissue reaction • Ladder-like DNA fragmentation • In vivo, individual cells appear affected • Cellular swelling • Membranes are broken • ATP is depleted • Cell lyses, eliciting an inflammatory reaction • DNA fragmentation is random, or smeared • In vivo, whole areas of the tissue are affected Necrosis Apoptosis (Richerd et.al., 2001)
    11. 11. 10/09/13 11 http://science.howstuffworks.com/life/cellular-microscopic/apoptosis.htm Fig -2 : Phases of PCD in a Vascular PlantFig -2 : Phases of PCD in a Vascular Plant
    12. 12. Why should a cell decide to commit suicide?Why should a cell decide to commit suicide? PCD is needed for proper development Examples:  Deletion of suspensor cells in embryos  Deletion of stamen primordia cells in unisexual flowers  The resorption of the tadpole tail  The formation of the fingers and toes of the fetus PCD is needed to destroy cells Examples: – Cells infected with viruses – Cells with DNA damage – Cancer cells – Galls 1210/09/13
    13. 13. PCD Regulators Caspase (Cysteinyl aspartate-specific proteases) Bcl-2 family Signal transduction pathway Caspase-8 Caspase-9 Caspase-3 Caspase-6 Caspase-7 Death initiators Effectors Bcl-2, Bcl-X (antiapoptotic) Bax, Bid and Bad (Promoters) Fas- ligand induced apoptosis (Jean et al., 1996, USA) 1310/09/13
    14. 14. 10/09/13 14 Programmed cell death
    15. 15. Programmed cell death : Pathways Death Ligands Effector Caspase 3 Death Receptors Initiator Caspase 8 PCDPCD Mitochondria/ Cytochrome C Initiator Caspase 9 “Extrinsic Pathway” “Intrinsic Pathway” DNA damage & p53DNA damage & p53 between G1between G1 →→ SS 1510/09/13
    16. 16. Fig-3 : Activation of apoptosis from outside the cell. 1610/09/13
    17. 17. APOPTOSIS: important in embryogenesis Morphogenesis (eliminates excess cells): Selection (eliminates non-functional cells): (Kerr et al., 1992, Germany) 1710/09/13
    18. 18. APOPTOSIS: important in embryogenesis Immunity (eliminates dangerous cells): Self antigen recognizing cell Organ size (eliminates excess cells): (Kerr et al., 1992, Germany)1810/09/13
    19. 19. APOPTOSIS: important in adults Tissue remodeling (eliminates cells no longer needed): Virgin mammary gland Late pregnancy, lactation Involution (non-pregnant, non-lactating) Apoptosis Apoptosis - Testosterone Prostate gland (Kerr et al., 1992, Germany)
    20. 20. Fig -4 : Sites of PCD in a Vascular Plant. The orange spheres represent internal dead cells, and the branched structures on the leaves represent trichomes (Roger et al., 1997, California) Fig -4 : Sites of PCD in a Vascular Plant. The orange spheres represent internal dead cells, and the branched structures on the leaves represent trichomes (Roger et al., 1997, California)
    21. 21. Fig -5 : Functions of PCD in Plants. The red regions represent cells that have been targeted for PCD and the orange regions represent cells that have died by PCD. Fig -5 : Functions of PCD in Plants. The red regions represent cells that have been targeted for PCD and the orange regions represent cells that have died by PCD. Deletion of suspensor cells in embryos Deletion of stamen primordia cells in unisexual flowers Deletion of root cap cells Deletion of leaf cells during leaf lobing Deletion of cells during cell specialization in TEs cells in an HR cells in uninfected leaves in response to HR-derived signals Deletion of aleuron cells in seeds (Roger et al., 1997, California)10/09/13
    22. 22. Aleuron layer In seeds , aleuron cells form a secretary tissue that releases hydrolases to digest the endosperm and nourish the embryo. Unnecessary Die (Kuo et al., 1996)
    23. 23. Root Cap CellsRoot Cap Cells Cell death occurs in root caps when roots are grown,Cell death occurs in root caps when roots are grown, showing that cell death is a normal part of development andshowing that cell death is a normal part of development and notnot a consequence ofa consequence of abrasionabrasion during soil penetration.during soil penetration. Shrink,Shrink, DNA staining - nuclei - becomeDNA staining - nuclei - become condensed.condensed. (Wang(Wang et alet al., 1996)., 1996)
    24. 24. (A) Electron micrograph of a living cell. (B) Electron micrograph of a dead cell. In the dead cell, the plasma membrane (pm) is separated from the cell wall (cw), and the protoplast is shrunken and condensed. Fig-6 : PCD in an Embryogenic Suspension Culture of Carrot (Roger et al., 1997, California)
    25. 25. Senescence  Senescence is the final phase of plant vegetative and reproductive development, preceding the widespread death of cells and organs.  Involves the active turnover and recapture of cellular material for use in other organs.  Membrane integrity maintained until late into the senescence process, suggesting that there is little or no leakage of cellular contents. (Nooden, 1988)
    26. 26. 10/09/13 26 Vascular plants transport water in columns of specialized dead cells termed TEs. Differentiation of TEs involves cell elongation, the deposition of cell wall components, including lignin, and autolysis. Autolysis begins as the cytoplasm and nuclei become lobed, condensed, and shrunken and ends as the cytoplasm breaks into small packets Xylogenesis- TE’sXylogenesis- TE’s (Roger et al., 1997, California)
    27. 27. INTERACTIONS WlTH THE ENVIRONMENT Hypoxia (O2 deprivation)  Cell death can occur in the cortex of the root and stem base in response to water logging and hypoxia.  The hypoxia leads to ROS (H2O2, Ethylene)  Ethylene – trigger apoptotic path way - eliminate some cells- aerenchyma.  The interna1 air spaces generated by cell death facilitate more efficient transfer of O2 from aerial organs to waterlogged stem bases and roots. (Armstrong, 1979)
    28. 28. 10/09/13 28 Plant-Pathogen interactions Plants can recognize certain pathogens and activate defenses (called the resistance response) that result in the limitation of pathogen growth at the site of infection. One dramatic hallmark of the resistance response is the induction of a localized cell death response (the hypersensitive response or HR) at the site of the infection. The HR is likely to be important for limiting a pathogen's nutrient supply, since the dying tissue rapidly becomes dehydrated. The HR appears to be a form of PCD in plants. since apoptotic features such as DNA breaks with 3'OH ends, blebbing of the plasma membrane as well as nuclear and cytoplasmic condensation are present in some cells undergoing the HR. (Jean et al., 1996, USA)
    29. 29. Case studies 2910/09/13
    30. 30. Tomato: Lycopersicon esculantum cv. Zhonghusihao Grown in GH 260 C / 180 C , day/ night and natural day length and stored @ 250 C . Devided in to groups (5 in each). In the heat shock treatment, fruit bag sub merged in water bath for 20 min @ 450 C, the control was placed @ 250 C for 20 min. Objectives: To identify which caspase- like enzymes are induced in the tomato fruit heat stress response and to monitor the effect of caspase like –proteases on heat stress- induced cell death.
    31. 31. Fig-7: Measurement of relative ion leakage from pericarp discs of tomato fruit as an indicator of cell death by measuring ion leakage from pericarp discs. (Gui et al., 2009,U.S.A.) Fig-7: Measurement of relative ion leakage from pericarp discs of tomato fruit as an indicator of cell death by measuring ion leakage from pericarp discs. (Gui et al., 2009,U.S.A.)
    32. 32. Fig-8: In-situ detection of DNA fragmentation by TUNEL analysis in tomato fruit. (Gui et al., 2009, U.S.A) a–c Pericarp sample from control tomato fruit. d–f Pericarp sample from tomato fruit that was treated at 45°C for 20 min g–i Pericarp sample from tomato fruit 6 h after treatment at 45°C for 20 min. j–l Pericarp sample from tomato fruit 12 h after treatment at 45°C for 20 min. Samples were stained using the TUNEL method (a, d, g, j), DAPI (b, e, h, k) bright-field (c, f, i, l). Arrows indicate TUNEL-positive nuclei.
    33. 33. Fig-9 : In-situ detection of DNA fragmentation by TUNEL analysis in tomato fruit. (Gui et al., 2009, U.S.A)10/09/13
    34. 34. Caspase-1 (Ac-YVAD-AMC), caspase-3 (Ac-DEVD-AMC), caspase-8 (Ac-IETD- AMC) and caspase-9 (Ac-LEHD-AMC) , HS - Treated at 45°C for 20 min, Fig-10 : Substrate specificity of HS-induced caspase-like activities in tomato fruit. (Gui et al., 2009, U.S.A)10/09/13
    35. 35. 3510/09/13 Fig -11 : Effect of Z-VAD-FMK pretreatment on cell death in heat treated tomato pericarp discs. . (Gui et al., 2009, U.S.A) Fig -11 : Effect of Z-VAD-FMK pretreatment on cell death in heat treated tomato pericarp discs. . (Gui et al., 2009, U.S.A) caspase-3 (Ac-DEVD-AMC), caspase-9 (Ac-LEHD-AMC) , Heat - Treated at 45°C for 20 min and Z-VAD-FMK (Z-Val-Ala-Asp Flouromethylkitone).
    36. 36. Materials and methods: Tomato ( Lycopersicon esculentum Mill.) cell suspension cultures, line Msk 8 (Koornneef et al., 1987) Murashige and Skoog liquid medium (5µMNAA, 1µM BA and vitamins) Cells were sub-cultured every 7 days and used for experiments 5 d after sub culture. Cell death inducers (Camptothecin-5µM, Staurosporin-2µM, Fumonisin-20µM) . 5% percent buffer formaline for detection of nuclear changes and DNA fragmentation. 36 Objectives: To study the PCD by treatment with known inducers of apoptosis in mammalian cells.
    37. 37. Fig -12 : Induction of cell death in tomato suspension cells by Fluorescine diacetate (FDA )staining method. □ - Staurosporin-2µM ∆ - Camptothecin-5µM X - Fumonisin-20µM O –Untreated control □ - Staurosporin-2µM ∆ - Camptothecin-5µM X - Fumonisin-20µM O –Untreated control (Anke et al., 2000, Netherland )
    38. 38. STAU- Staurosporin-2µM CPT - Camptothesin-5µM FUM - Fumonisin-20µM FeSo4 -100mM STAU- Staurosporin-2µM CPT - Camptothesin-5µM FUM - Fumonisin-20µM FeSo4 -100mM Fig-13: Chemical induced DNA fragmentation in tomato suspension cells by using DNA laddering pattern. (Anke et al., 2000, Netherland )10/09/13
    39. 39. 5µM -Camptothecin (CPT), 50µM- N-ethylmaleimide (NEM), 5µM -iodoacetamide (IA), 200µM- Nm Nα-p-tosyl- L-lysine chloromethylekitone (TLCK), 1mM -4,2,amiloethyle benzensulfonyl fluoride (AEBSF) 20µM -Fumonisin BI (FUM) 2µM -Staurosporine (STAU) 5µM -Camptothecin (CPT), 50µM- N-ethylmaleimide (NEM), 5µM -iodoacetamide (IA), 200µM- Nm Nα-p-tosyl- L-lysine chloromethylekitone (TLCK), 1mM -4,2,amiloethyle benzensulfonyl fluoride (AEBSF) 20µM -Fumonisin BI (FUM) 2µM -Staurosporine (STAU) Fig -14: Inhibition of chemical induced cell death in tomato suspension cells by protease inhibitors. (Anke et al., 2000, Netherland )39
    40. 40. CPT - Camptothesin-5µM (CPT+ YVAD) – 100nM Caspase specific peptide inhibitor- Acyl -Tyr- Val-Ala-Asp- chloromethylketone . (CPT+ AAPV) – 100nM control peptide - MethoxySuccinyl-Ala-Ala- Pro-Val-chloromethylketone . CPT - Camptothesin-5µM (CPT+ YVAD) – 100nM Caspase specific peptide inhibitor- Acyl -Tyr- Val-Ala-Asp- chloromethylketone . (CPT+ AAPV) – 100nM control peptide - MethoxySuccinyl-Ala-Ala- Pro-Val-chloromethylketone . Fig-15: Inhibition of DNA fragmentation by caspase specific peptide inhibitors by DNA laddering pattern. (Anke et al., 2000, Netherland )10/09/13
    41. 41. 4110/09/13 Tomato seeds were incubated in the dark @ 250 c prior to use. Isolation of endosperm tissue was performed aseptically and were surface sterilized for 10 s with 70% ethanol. Then endosperm halves were made and used for the experiment.
    42. 42. Fig -16 : Post-germinative tomato endosperm cells display features of reserve mobilization and PCD including SlCysEP-containing ricinosomes. (Christoper et al.,2013, Fig -16 : Post-germinative tomato endosperm cells display features of reserve mobilization and PCD including SlCysEP-containing ricinosomes. (Christoper et al.,2013, a) Cells farthest from the embryo show less reserve mobilization and no signs of PCD. Cells closer to the embryo contain fewer reserves; cells next to the embryo are crushed or have recently died b)The top cell farthest from the embryo appears disorganized suggesting it is undergoing PCD; cells closer to the embryo (below frame) appear dead with large lipid aggregates c) A cell showing signs of reserve mobilization, but not PCD, contains a developing ricinosomes like structure 4210/09/13
    43. 43. Conti…. d)Localization of SlCysEP within ricinosomes is evidenced by the white arrowheads lying within the ricinosomes, e) A labelled-ricinosome in a cell with few reserves next to a dead cell (top left) illustrating the asynchronous nature of endosperm PCD Fig -17 : Post-germinative tomato endosperm cells display features of reserve mobilization and PCD, including SlCysEP-containing ricinosomes. (Christoper et al.,2013, Heidelberg) Fig -17 : Post-germinative tomato endosperm cells display features of reserve mobilization and PCD, including SlCysEP-containing ricinosomes. (Christoper et al.,2013, Heidelberg)4310/09/13
    44. 44. Fig-18 : SlCysEp transcript and enzyme accumulation in tomato seeds. (Christoper et al.,2013, Heidelberg) Fig-18 : SlCysEp transcript and enzyme accumulation in tomato seeds. (Christoper et al.,2013, Heidelberg) 4410/09/13 HAI 34 k BP
    45. 45. Fig -19 : Tissue specific SlCysEP expression and enzyme accumulation using Northern-blot analysis (Christoper et al.,2013, Heidelberg) Fig -19 : Tissue specific SlCysEP expression and enzyme accumulation using Northern-blot analysis (Christoper et al.,2013, Heidelberg)4510/09/13
    46. 46. a) Non-germinated (NG), b) short seedling (SS), c) long seedling (LS) Endosperm halves from NG (d), SS , LS (f) seeds. (*) the micropylar endosperm collar Fig-20: SlCysEP accumulation pattern change in response to development @144HAI. (Christoper et al.,2013, Heidelberg) Fig-20: SlCysEP accumulation pattern change in response to development @144HAI. (Christoper et al.,2013, Heidelberg)4610/09/13
    47. 47. 4710/09/13 Fig-21 :Quantification of protien and SlCysEP profile isolated on en equal tissue basis. (Christoper et al.,2013, Fig-21 :Quantification of protien and SlCysEP profile isolated on en equal tissue basis. (Christoper et al.,2013,
    48. 48. Tomato (Lycopersicon esculentum Mill.) cell suspension culture, line Msk8, was grown in a liquid MS medium + vitamins and minerals, supplemented with 5 μM a-NAA, 1 μM 6-BA and 3% (w/v) sucrose . Cell death inducers CdSO4 and CdCl2 were added to 5 ml of suspension culture, Inhibitors were applied 30 min before CdSO4. In general, Inhibitors were tested in a range of concentrations (from nM to mM) with and without the addition of 100 μM CdSO4. Lowest concentrations of inhibitors giving significant reduction of CdSO4-induced cell death are presented. The next chemicals have been tested for their potency to suppress Cd-induced cell death: Cell death determination by fluorescence microscopy. Hydrogen peroxide assay Ethylene production measurement 4810/09/13 Biotechnology & Biotechnological Equipment, 2009 23(2): 538-541
    49. 49. Fig -22: Effect of cadmium salts on cell death in tomato suspension cells 24 h after the treatments. (Iakimova et al., 2009, Bulgaria ) Fig -22: Effect of cadmium salts on cell death in tomato suspension cells 24 h after the treatments. (Iakimova et al., 2009, Bulgaria ) 4910/09/13
    50. 50. Fig -23: Morphological features of living (A) and dead (B) tomato cells 24 h after treatment with 100 μM CdSO4. ( Iakimova et al., 2009, Bulgaria ) Fig -23: Morphological features of living (A) and dead (B) tomato cells 24 h after treatment with 100 μM CdSO4. ( Iakimova et al., 2009, Bulgaria ) A - living cell with diffused nucleus, preserved cytoskeleton and intact cytoplasm; B – dead cell showing compacted nucleus and shrunken cytoplasm separated from the cell wall. cyt-cytoplasm, cw- cell wall, nu- nucleus. 5010/09/13
    51. 51. Fig-24: Kinetic of hydrogen peroxide production in tomato suspension cells exposed to 100 μM CdSO4. ( Iakimova et al., 2009, Bulgaria ) Fig-24: Kinetic of hydrogen peroxide production in tomato suspension cells exposed to 100 μM CdSO4. ( Iakimova et al., 2009, Bulgaria ) 5110/09/13
    52. 52. Fig -25: Kinetic of ethylene production in tomato suspension cells exposed to 100 μM CdSO4. ( Iakimova et al., 2009, Bulgaria ) Fig -25: Kinetic of ethylene production in tomato suspension cells exposed to 100 μM CdSO4. ( Iakimova et al., 2009, Bulgaria ) 5210/09/13
    53. 53. Objectives : Elicitors of different origin (fumonisin B1, fungal toxin), camptothecin (alkaloid from Camptotheca acuminata), mastoparan (wasp venom) and the heavy metal (cadmium) were tested for their ability to induce programmed cell death (PCD) in a model system of tomato cell culture, line MsK8. 5310/09/13
    54. 54. Fig-26 : Effect of 20 μM Fumonisin B1 (FumB1), 5 μM camptothecin (CPT), 100 μM CdSO4 or 5 μM mastoparan (MP) on cell death induction in tomato suspension cells. (Elena et al., 2007, Bulgaria) Fig-26 : Effect of 20 μM Fumonisin B1 (FumB1), 5 μM camptothecin (CPT), 100 μM CdSO4 or 5 μM mastoparan (MP) on cell death induction in tomato suspension cells. (Elena et al., 2007, Bulgaria) 5410/09/13
    55. 55. Fig -27 : Effect of 100 nM Ac-YVAD-CMK, 100 nM Z-Asp-CH2-DCB and 1mM (AEBSF) on cell death in tomato suspension cells induced by 20 μM FumB1, 5 μM CPT, 100 μM CdSO4 or 5 μM MP. Cell death was calculated after 24 h of treatment followed by FDA staining of the living cells and is expressed as per cent of inhibition. Fig -27 : Effect of 100 nM Ac-YVAD-CMK, 100 nM Z-Asp-CH2-DCB and 1mM (AEBSF) on cell death in tomato suspension cells induced by 20 μM FumB1, 5 μM CPT, 100 μM CdSO4 or 5 μM MP. Cell death was calculated after 24 h of treatment followed by FDA staining of the living cells and is expressed as per cent of inhibition. 5510/09/13
    56. 56. 5610/09/13  PCD is essential for ensuring the proper development of plant  Interaction with environment which results robust defence response against biotic and abiotic stresses
    57. 57. 10/09/13 57 GERMAN SCIENTIST CARL VOGT WAS FIRST TO DESCRIBE THE PRINCIPLE OF APOPTOSIS IN 1842. GERMAN SCIENTIST CARL VOGT WAS FIRST TO DESCRIBE THE PRINCIPLE OF APOPTOSIS IN 1842. IN 1885, ANATOMIST WALTHER FLEMMING DELIVERED A MORE PRECISE DESCRIPTION OF THE PROCESS OF PROGRAMMED CELL DEATH. JONE E. SULSTON WON THE NOBEL PRIZE IN MEDICIN IN 2002 FOR HIS POINERING RESRACH ON APOPTOSIS JONE E. SULSTON WON THE NOBEL PRIZE IN MEDICIN IN 2002 FOR HIS POINERING RESRACH ON APOPTOSIS

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