Changes in DNA synthesis during cell growth and chloroplast replication in greening spinach leaf disks

Abstract

Spinach leaf disks grown initially in the dark, show increased cell expansion and chloroplast replication when transferred to the light. These changes are accompanied by increases in the total amount of DNA and the incorporation of [3H]thymidine (3H-TdR). Autoradiography of EDTA-separated cells dried on to glass slides was used to follow changes in 3H-TdR incorporation in both chloroplasts and nuclei. Specificity of incorporation was confirmed by nuclease studies. DNA synthesis occurs in both the chloroplasts and nuclei, and is highest just prior to, and during the period of most rapid cell growth and chloroplast replication which occurs shortly after the transfer to the light. Light, however, appears to have a greater and more immediate effect on nuclear DNA synthesis. Though nuclear and chloroplast DNA syntheses follow similar patterns during disk growth, in a given cell, chloroplast DNA synthesis can be separate in time from nuclear DNA synthesis. The increased nuclear DNA synthesis is possibly required to support the increased population of chloroplasts, while chloroplast DNA synthesis is associated with chloroplast division. If the disks are not transferred to the light but kept in darkness, chloroplast 3H-TdR incorporation remains high, though chloroplast division is reduced. Epidermal cells in light-grown tissue also show 3H-TdR incorporation but low rates of chloroplast division. It would appear that chloroplast DNA synthesis in mesophyll cells from light-grown tissue shows a general relation to chloroplast division, but there does not appear to be an obligatory close coupling between the 2 processes.