Diurnal Changes in Microscopic Structures of Mesophyll Cells of Norway Spruce, Picea abies(L.) Karst., and the Effects of Ozone and Drought

Microscopy has been used to diagnose the effects of air pollutantson conifers. As sampling may be slow, it is important to knowwhether the structure of needles changes during the day. Usinglight- and electron microscopy, diurnal changes in the structureof mesophyll cells of current year needles from 6-year-old Norwayspruce saplings were studied on one day in early autumn, 1995.Saplings had been grown in open-top chambers and exposed toozone and drought stress during the 1992–1995 growingseasons. In all treatments, the proportion of cells with smallcytoplasmic vacuoles and with large cytoplasmic lipid depositsdecreased during the day. Mitochondria increased in size fromearly morning to midday, after which time they shrank progressively.Ozone affected the shape of mitochondria: the proportion oftube-shaped mitochondria decreased from early morning in controlsaplings, whereas it increased in ozone-treated saplings. Starchaccumulated during the day, but to a lesser extent in drought-stressedsaplings than well-watered controls. Microbodies were smallerin drought-stressed saplings than in controls early in the morning,but their size then increased compared to that of the controls.Cellular and sub-cellular disorganization, probably due to sub-optimalfixation procedures, were detected in all treatments, especiallyin the morning, but these decreased later, particularly in well-wateredsaplings. Results show that microscopy is a sensitive and validtool for studying stress responses in plants. However, the timeof sample collection must be considered and documented to avoidmisleading conclusions due to actual diurnal changes and fixationproblems. Copyright 2001 Annals of Botany Company Diurnal changes, drought, light microscopy, mesophyll cells, needles, Norway spruce, Picea abies, ozone, transmission electron microscopy