Changes in cell cycle status and expression of p34cdc2 kinase during potato tuber meristem dormancy

Michael A. Campbell, Jeffrey C. Suttle, Thomas W. Sell

Research output: Contribution to journalArticle

39 Citations (Scopus)

Abstract

Potato (Solanum tuberosum L. cv. Russet Burbank) tubers undergo a period of endodormancy that is characterized by cell division arrest. At the time of harvest, the tubers used in this study were completely endodormant (i.e. 0% sprouting). After 120 days of storage at 3°C, tubers transferred to 20°C had begun to exit endodormancy and exhibited ca 50% sprouting. After 223 days of 3°C storage, tubers transferred to 20°C were completely nondormant and exhibited 100% sprouting. Based on flow cytometry, about 70% of nuclei isolated from endodormant meristems are arrested in the G1/G0 phase of the cell cycle. Storage of tubers at 3°C did not alter the cell cycle position nor did transfer of tubers from 3 to 20°C for 7 days prior to analysis unless tubers had been stored for at least 223 days. After 223 days of cold (3°C) storage, tubers transferred to 20°C for 7 days showed sprout growth in excess of 5 mm and an increase in the percentage of nuclei in the G2 phase of the cell cycle. Uptake and incorporation of 3H-thymidine into DNA was low in all tubers up until 120 days postharvest. After that time, only tubers incubated at 20°C for 7 days prior to analysis exhibited an increase in 3H-thymidine incorporation. This increase coincided with visible sprout growth, demonstrating that cell cycle shifts in tuber meristems relate directly to sprout growth and not the breakage of the endodormancy per se. Using degenerate primers, a portion of a p34cdc2 homolog was amplified from RNA isolated from log-phase potato suspension culture cells by polymerase chain reaction. Northern analysis with this probe demonstrated that mRNA levels for two p34cdc2 homologs were present throughout the endodormant period. Imrnunoblot analysis demonstrated that levels of at least four proteins containing a PSTAIRE epitope (i.e. cdc2-like) were present at reduced levels in endodormant meristems and increased in reactivated tuber meristems that showed a shift in cell cycle kinetics based on flow cytometry and increased 3H-thymidine incorporation. These results indicate that the temporal shift in competence for cell division in potato meristems induced by dormancy is not accompanied by alterations in the level of mRNA for p34cdc2 homologues but is correlated with a change in the level of PSTAIRE-containing proteins. This suggests that during endodormancy cell division in potato tuber meristems is regulated indirectly by post-transcriptional regulation of genes controlling the cell cycle.

Original languageEnglish (US)
Pages (from-to)743-752
Number of pages10
JournalPhysiologia Plantarum
Volume98
Issue number4
DOIs
StatePublished - Jan 1 1996

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Meristem
Solanum tuberosum
meristems
dormancy
cell cycle
Cell Cycle
tubers
phosphotransferases (kinases)
Phosphotransferases
potatoes
Cell Division
Thymidine
Flow Cytometry
Growth
thymidine
sprouting
cdc Genes
Cell Cycle Resting Phase
Messenger RNA
cell division

All Science Journal Classification (ASJC) codes

  • Physiology
  • Genetics
  • Plant Science
  • Cell Biology

Cite this

@article{d14aab20f1384c94b503f1867608c8a6,
title = "Changes in cell cycle status and expression of p34cdc2 kinase during potato tuber meristem dormancy",
abstract = "Potato (Solanum tuberosum L. cv. Russet Burbank) tubers undergo a period of endodormancy that is characterized by cell division arrest. At the time of harvest, the tubers used in this study were completely endodormant (i.e. 0{\%} sprouting). After 120 days of storage at 3°C, tubers transferred to 20°C had begun to exit endodormancy and exhibited ca 50{\%} sprouting. After 223 days of 3°C storage, tubers transferred to 20°C were completely nondormant and exhibited 100{\%} sprouting. Based on flow cytometry, about 70{\%} of nuclei isolated from endodormant meristems are arrested in the G1/G0 phase of the cell cycle. Storage of tubers at 3°C did not alter the cell cycle position nor did transfer of tubers from 3 to 20°C for 7 days prior to analysis unless tubers had been stored for at least 223 days. After 223 days of cold (3°C) storage, tubers transferred to 20°C for 7 days showed sprout growth in excess of 5 mm and an increase in the percentage of nuclei in the G2 phase of the cell cycle. Uptake and incorporation of 3H-thymidine into DNA was low in all tubers up until 120 days postharvest. After that time, only tubers incubated at 20°C for 7 days prior to analysis exhibited an increase in 3H-thymidine incorporation. This increase coincided with visible sprout growth, demonstrating that cell cycle shifts in tuber meristems relate directly to sprout growth and not the breakage of the endodormancy per se. Using degenerate primers, a portion of a p34cdc2 homolog was amplified from RNA isolated from log-phase potato suspension culture cells by polymerase chain reaction. Northern analysis with this probe demonstrated that mRNA levels for two p34cdc2 homologs were present throughout the endodormant period. Imrnunoblot analysis demonstrated that levels of at least four proteins containing a PSTAIRE epitope (i.e. cdc2-like) were present at reduced levels in endodormant meristems and increased in reactivated tuber meristems that showed a shift in cell cycle kinetics based on flow cytometry and increased 3H-thymidine incorporation. These results indicate that the temporal shift in competence for cell division in potato meristems induced by dormancy is not accompanied by alterations in the level of mRNA for p34cdc2 homologues but is correlated with a change in the level of PSTAIRE-containing proteins. This suggests that during endodormancy cell division in potato tuber meristems is regulated indirectly by post-transcriptional regulation of genes controlling the cell cycle.",
author = "Campbell, {Michael A.} and Suttle, {Jeffrey C.} and Sell, {Thomas W.}",
year = "1996",
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Changes in cell cycle status and expression of p34cdc2 kinase during potato tuber meristem dormancy. / Campbell, Michael A.; Suttle, Jeffrey C.; Sell, Thomas W.

In: Physiologia Plantarum, Vol. 98, No. 4, 01.01.1996, p. 743-752.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Changes in cell cycle status and expression of p34cdc2 kinase during potato tuber meristem dormancy

AU - Campbell, Michael A.

AU - Suttle, Jeffrey C.

AU - Sell, Thomas W.

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Y1 - 1996/1/1

N2 - Potato (Solanum tuberosum L. cv. Russet Burbank) tubers undergo a period of endodormancy that is characterized by cell division arrest. At the time of harvest, the tubers used in this study were completely endodormant (i.e. 0% sprouting). After 120 days of storage at 3°C, tubers transferred to 20°C had begun to exit endodormancy and exhibited ca 50% sprouting. After 223 days of 3°C storage, tubers transferred to 20°C were completely nondormant and exhibited 100% sprouting. Based on flow cytometry, about 70% of nuclei isolated from endodormant meristems are arrested in the G1/G0 phase of the cell cycle. Storage of tubers at 3°C did not alter the cell cycle position nor did transfer of tubers from 3 to 20°C for 7 days prior to analysis unless tubers had been stored for at least 223 days. After 223 days of cold (3°C) storage, tubers transferred to 20°C for 7 days showed sprout growth in excess of 5 mm and an increase in the percentage of nuclei in the G2 phase of the cell cycle. Uptake and incorporation of 3H-thymidine into DNA was low in all tubers up until 120 days postharvest. After that time, only tubers incubated at 20°C for 7 days prior to analysis exhibited an increase in 3H-thymidine incorporation. This increase coincided with visible sprout growth, demonstrating that cell cycle shifts in tuber meristems relate directly to sprout growth and not the breakage of the endodormancy per se. Using degenerate primers, a portion of a p34cdc2 homolog was amplified from RNA isolated from log-phase potato suspension culture cells by polymerase chain reaction. Northern analysis with this probe demonstrated that mRNA levels for two p34cdc2 homologs were present throughout the endodormant period. Imrnunoblot analysis demonstrated that levels of at least four proteins containing a PSTAIRE epitope (i.e. cdc2-like) were present at reduced levels in endodormant meristems and increased in reactivated tuber meristems that showed a shift in cell cycle kinetics based on flow cytometry and increased 3H-thymidine incorporation. These results indicate that the temporal shift in competence for cell division in potato meristems induced by dormancy is not accompanied by alterations in the level of mRNA for p34cdc2 homologues but is correlated with a change in the level of PSTAIRE-containing proteins. This suggests that during endodormancy cell division in potato tuber meristems is regulated indirectly by post-transcriptional regulation of genes controlling the cell cycle.

AB - Potato (Solanum tuberosum L. cv. Russet Burbank) tubers undergo a period of endodormancy that is characterized by cell division arrest. At the time of harvest, the tubers used in this study were completely endodormant (i.e. 0% sprouting). After 120 days of storage at 3°C, tubers transferred to 20°C had begun to exit endodormancy and exhibited ca 50% sprouting. After 223 days of 3°C storage, tubers transferred to 20°C were completely nondormant and exhibited 100% sprouting. Based on flow cytometry, about 70% of nuclei isolated from endodormant meristems are arrested in the G1/G0 phase of the cell cycle. Storage of tubers at 3°C did not alter the cell cycle position nor did transfer of tubers from 3 to 20°C for 7 days prior to analysis unless tubers had been stored for at least 223 days. After 223 days of cold (3°C) storage, tubers transferred to 20°C for 7 days showed sprout growth in excess of 5 mm and an increase in the percentage of nuclei in the G2 phase of the cell cycle. Uptake and incorporation of 3H-thymidine into DNA was low in all tubers up until 120 days postharvest. After that time, only tubers incubated at 20°C for 7 days prior to analysis exhibited an increase in 3H-thymidine incorporation. This increase coincided with visible sprout growth, demonstrating that cell cycle shifts in tuber meristems relate directly to sprout growth and not the breakage of the endodormancy per se. Using degenerate primers, a portion of a p34cdc2 homolog was amplified from RNA isolated from log-phase potato suspension culture cells by polymerase chain reaction. Northern analysis with this probe demonstrated that mRNA levels for two p34cdc2 homologs were present throughout the endodormant period. Imrnunoblot analysis demonstrated that levels of at least four proteins containing a PSTAIRE epitope (i.e. cdc2-like) were present at reduced levels in endodormant meristems and increased in reactivated tuber meristems that showed a shift in cell cycle kinetics based on flow cytometry and increased 3H-thymidine incorporation. These results indicate that the temporal shift in competence for cell division in potato meristems induced by dormancy is not accompanied by alterations in the level of mRNA for p34cdc2 homologues but is correlated with a change in the level of PSTAIRE-containing proteins. This suggests that during endodormancy cell division in potato tuber meristems is regulated indirectly by post-transcriptional regulation of genes controlling the cell cycle.

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