Zbornik gozdarstva in lesarstva, 39, 1992, s. 117 - 132 
FDC: 174.7 Picea abies (L.) Karst.: 164.5:416.1:161.32+160.201 (497.12 TEŠ) 
MORPHOLOGICAL AND ANATOMICAL CHANGES OF NORWAY 
SPRUCE NEEDLES (Picea abies (L.) Karst.) IN THE ŠOŠTANJ 
STEAM POWER PLANT INFLUENCE AREA 
Alenka KMECL*, Franc BATIČ** 
Abstract 
Morphological and anatomical changes of Norway spruce needles from 16 sampling 
plots in the vicinity of the Steam Power Plant in Šoštanj (TEŠ) were examined. The 
visible needle injuries were analysed and compared to the sulphur contents in the 
same samples. On the basis of this comparison and analysis of needle infections by 
microorganisms a conclusion was made on the possible causes of needle injuries. A 
new method of evaluation of needle injuries was tested. The method was based on 
autofluorescence of chlorophyll in spruce needles. After DCMU treatment of needle 
samples, the fluorescence intensity rose significantly, yet not proportionally to the 
autofluorescence intensity. The difference between the autofluorescence and the 
secondary fluorescence intensity (after DCMU treatment) showed, to which degree 
was damaged the photosynthetic apparatus of the Norway spruce-needles. 
Key words: Norway spruce, needles, injuries, autofluorcsccnce of chlorophyll, 
DCMU, secondary fluorescence of chlorophyll 
MORFOLOŠKE IN ANATOMSKE SPREMEMBE SMREKOVIH IGLIC 
Picea abies (L.) Karsten V VPLIVNEM OBMOČJU 
TERMOELEKTRARNE ŠOŠTANJ 
Izvleček 
Ugotavljali smo morfološke in anatomske spremembe smrekovih iglic s 16 vzorčnih 
mest v okolici TEŠ. Poškodovanost iglic smo primerjali z vsebnostjo žvepla v istih 
vzorcih. Na podlagi omenjene primerjave in analize okuženosti iglic z 
mikroorganizmi smo sklepali o izvoru njihovih poškodb. Preizkusili smo novo metodo 
vrednotenja poškodovanosti iglic na osnovi avtofluorescence klorofila v njih. Po 
dodatku DCMU se je fluorescenca močno povečala, vendar ne sorazmerno z 
avtoflorescenco. Razlika v intenzivnosti avtofluorescence in sekundarne fluorescence 
pokaže, v kolikšni meri je poškodovan fotosintetski aparat pri rastlini. 
Ključne besede: Picea abies, iglice, poškodovanost, avtofluorescenca klorofila, DCMU 
(3 3,4 diklorfenil - 1,1 dimetil urea), sekundarna fluorescenca klorofila 
A. K. Dipl. Biol., Zurich 
Prof. Dr. F. B., Dipl. Biol., Inštitut za gozdno in lesno gospodarstvo, Večna pol 2, 61000 Ljubljana, Slovenija 
Zbornik gozdarstva in lesarstva, 39 
CONTENTS 
1 INTRODUCTION 119 
2 MATERIAL 119 
3 MORPHOLOGICAL ANALYSIS 121 
3.1 Methods 121 
3.2 Results 121 
4 ANALYSIS OF NEEDLE INFECTIONS BY MICROORGANISMS 123 
4.1 Methods 123 
4.2 Results 123 
5 ANALYSIS OF CHLOROPLAST FLUORESCENCE 124 
5.1 Methods 124 
5.2 Results 125 
6 DISCUSSION 127 
7 LITERATURE 128 
118 
Kmecl A., Batič F., Morfological and anatomical.. 
1 INTRODUCTION 
Forest decline has recently been considered one of the major ecological problems of 
the developed countries. Slovenia had to face this problem similarly as almost every 
other European country - due to its geographical position and developmental level. 
The first systematic inventory of forest decline in Slovenia (in 1985) showed, that the 
situation was alarming. The greatest area of declining forest was found to be the 
valley of Šaleška dolina and its surroundings. 
The steam power plant in Šoštanj (TEŠ) is the greatest source of air pollution in 
Slovenia, and the relief of the valley of Šaleška dolina contributes greatly to the bad 
pollution circumstances. The local inventorization of 10140ha of forest showed, that 
98% of the forest had already been damaged (HRČEK et ali. 1988). 
Because of the worrying results of the forest decline inventory, numerous 
investigations of air pollution and damage to forests in Šaleška dolina were 
undertaken. Our investigation was a part of this wide research project, and 
contributed some data and results to it. Beside this, a new method of evaluation of 
Nmway spruce needle injuries was tested. The method was based on the difference 
of autofluorescence and secondary fluorescence of chlorophyll in differently injured 
needles, and was expected to show indirectly the quantity of chlorophyl in the Nor-
way spruce needles. 
It has been possible to test "our" methods of evaluation of chlorophyl quantity, be-
cause at the same tirne a biochemical investigation of the chlorophyl contents has 
been carried out and we could compare the results of the latter investigation (courte-
sy of Missis RIBARIČ - LASNIK - see Appendix 3) with the results of ours. 
In this article, much less attention is dedicated to the other two parts of 
investigation - to the morphological analysis of the needles and to the analysis of 
needle infections by microorganisms as this was routine work and has been exactly 
described elsewhere (ŠLIBAR 1990). · 
2 MATERIAL 
Norway spruce needles for the investigation were sampled at the same tirne and from 
the same trees as the needles, use.d for the analyses of chlorophyll and sulphur 
contents. Therefore the comparison of results of ali the three analyses could give an 
answer about the influence of air pollution on Norway spruce trees. 
The goal of this study was to analyse the injuries of needles from the following 16 
1 ocali ti es: 
119 
Zbornik gozdarstva in lesarstva, 39 
Sampling localities Height above Distance from Exposition 
sea leve! (m) TEŠ (m) 
Vrh Smrekovca 1577 12.750 NW 
Koča na Smrekovcu 1400 12.700 NW 
Kramarice 1200 12.750 NW 
Slanica 950 9.500 NW 
Zavodnje 770 8.250 NW 
Pod Zavodnjami 690 6.000 NW 
Lajše 400 2.000 NW 
Lokovica 505 500 sw 
Veliki vrh 560 2.750 SE 
Andraž 410 6.000 SE 
Graška gora 770 7.200 NE 
Šmiklavž 485 10.500 NE 
Podgorica 460 11.000 NE 
Kope 1542 20.200 NE 
Komisija 1300 21.750 NE 
Paški Kozjak 1108 10.500 NE 
The Šoštanj Steam Power Plant (TEŠ) is situated at height of 360m above sea leve!, 
at the bottom of the valley of Šaleška dolina. 
The sampling plots were located at various distances from TEŠ, at different altitudes 
and in variously damaged regions of forest. On each sampling plot, the most vita! 
two Norway spruce trees of the age of 60-80 years were chosen for the investigation. 
From these trees, the current year needles and the one year old needles were 
sampled from twigs of the seventh spruce whirl, counted from the tree crown top 
downwards. The sampling took place five times: in spring, summer and autumn 1989 
and in winter and spring 1990. 
120 
Kmecl A., Batič F., Morfological and anatomical.. 
3 MORPHOLOGICAL ANALYSIS 
3.1 Methods 
From each tree, 60 needles of the mentioned two age classes were analysed micro-
scopically (stereomicroscope ZEISS, JENA). The injured surface of every needle was 
evaluated and the data were registered. Later, the total damaged surface was 
summed for each group of 60 needles. Among injuries, only the necroses and 
chloroses were taken into account. The mechanical injuries were not taken into 
consideration, because they did not result from the influence of gas pollutants. 
However, the mechanical injuries occured only seldom. 
After assessment of injuries, the needles were dried until no further loss of weight 
could be noticed and then weighcd. The data on the dry weight of needles were used 
for comparison of needle injuries, originating from different sampling plots. 
3.2 Results 
The results are presented in figures 1 and 2. 
lID 
'Xl 
~oo 
~ 
tl 'iU 
:;; 
~m 
] 50 
:~ 
..... 40 
o 
,.830 
e 
:::, 
"'~ 
10 --two-year needles 
o 
.,i 
e e 
(/) 
~ 
.,i " "' " -d >~ tl ~ 
>N 
~ 
>~ "' 8.. u o '8' !i1 
., 
·~ e ·5 ·a .,, 3' ] ,a "' :;! e "' o " ~ :;;i t ~ e vi :;; N ~ ·s ., .s >U ~ N 1 "" o ..,. 
Figure 1: Comparison of injuries of current-year and one-year old needles 
(autumn, 1989) 
121 
.. ~ :;:::.o 
.!/) .... 
e ~ o :..: i,; 
Zbornik gozdarstva in lesarstva, 39 
The comparison of injuries of current-year and one-year old needles is shown, 
according to the number of injured needles to a needle-group (the 60 needles of the 
same age ). It could be seen from the diagram, that in the majority of cases the cur-
rent-year needles are less damaged than the one-year needles. The difference in 
damage among sampling plots varies significantly. The highest degree of injury oc-
curs by needles from the localities of higher altitude, although the sulphur contents 
in these same needles is the lowest ( courtesy of Mr. Kalan - see Appendix 1 ). 
4!XXl 
500 
-E 
> 
o 
'i:' 
-o 
o ;; 
N 
"O ~ 
~ 5 "' N 
-o o 
o. 
--one-year needles 
--0--two-year needles 
.. .C >N C! >N -~ 8.. "' "' 4 C! ;; '.[ ·,; > -o c3 .. ~ ~ C ::;;; o c5 ·a .. 3 -;; < -o ~ > >{/J o P-. 
Figure 2: Comparison of injuries of current-year needles and one-year old needles 
(autumn, 1989) 
~ . .., 
~ 
i,.; 
The comparison of injuries of current-year and one-year old needles from different 
sampling plots is shown as a quotient: injured surface/dry weight. Except for one 
case, the current-year needles are less injured than the one-year old needles, and 
again the extent of injuries varies significantly from one locality to another. 
The course of the curves on both diagrams is surprisingly similar. It had been 
expected that after dry weight of needles was taken into consideration, the relations 
between sampling plots would change significantly - yet this had not happened. 
Neither in Figure 2, the connection between external damage to needles and the 
contents of sulphur could be noticed. 
It is interesting to compare our results with the results of the analysis of sulphur 
contents in Norway spruce needles (KALAN). For this reason, the diagram of 
sulphur contents has also been appended to the article (Appendix 1). 
122 
Kmecl A., Batič F., Morfological and anatomical.. 
The analysis of external injuries of the Norway spruce needles of different age 
confirmed, that the current year needles were Iess damaged than the older ones. This 
had been expected, because the injuries increase with the longer tirne of exposure to 
damaging substances. 
In comparison of injuries to needles from various sampling plots it was expected, that 
the visible injuries would mainly depend on the sulphur contents in the needles. This 
proved not to be trne. Far the most injured were the needles from higher elevations, 
in which the sulphur contents was rather low. These injuries could therefore not be a 
result of damaging effects of sulphur, and the cause should be sought elsewhere. As 
high concentrations of ozone have often been measured in the surroundings of TEŠ 
it is possible for this gas to be the major pollutant in this case. lts concentrations 
increase with the growing altitude - probably as a consequence of greater number of 
days without fog, stronger UV radiation and possibly also the vicinity of stratosphere 
(which makes passing of the ozone to the lower air layers easier). Therefore the 
trees' injuries on these elevations are most possibly caused by ozone. 
4 ANALYSIS OF NEEDLE INFECTIONS BY 
MICROORGANISMS 
4.1 Methods 
Parts of fresh needles with different types of injuries were sterilized and inoculated 
on nutrient medium. Some days Iater the colonies were sterilly transfered to fresh 
media. After four weeks of incubation, the species of fungi were determined 
(ŠLIBAR 1990). 
4.2 Results 
10 types of micelia grew from the needles with various types of injuries. The micelia 
were not characteristical for different types of injuries - no connection was 
discovered between the type of injury and the fungi species. Ali the determined 
species of fungi were saprofitic, with an exception of the Sirococcus strobilinus 
Preuss (a parasitic species). Therefore it could be infered, that needle injuries did 
not occur as a consequence of infection. Most probably the microorganisms infected 
the already feeble needles. No hiphae or sporangies were noticed in the needle 
tissue, so that it could also be possible, that the presence of fungi on the needle 
surface was merely casual. It has been anticipated, that the only parasitic fungus, 
Sirococcus strobilinus Preuss, had not led to extensive needle injuries, for it only 
appeared once among the 75 inoculated samples (ŠLIBAR 1990). 
123 
Zbornik gozdarstva in lesarstva, 39 
5 ANALYSIS OF CHLOROPLAST FLUORESCENCE 
5.1 Methods 
If chlorophyll is exposed to UV light, strong red fluorescence appears. As a 
consequence of chlorophyll degradation, the red colour changes to orange-yellow. 
The healthy needles fluorescred, the chlorotic needles orange-yellow and the necrotic 
needles have no fluorescence at ali. The fluorescence intensity depends on the 
quantity of chlorophyll in the needles. 
By intensive illumination with UV light, the autofluorescence fades away very 
quickly. The discolouration can be delayed, if a drop of 3 (3,4dichloro-phenil)-1,1 
dimethyl urea (DCMU) solution is added to the sample instead of water. The 
fluorescent colour is blood-red at the beginning and soon changes to lighter red, 
which is more persistent. 
DCMU is a herbicide that specifically blocks the photosystem II (DENFFER, 
ZIEGLER 1982). Due to the blockade, the resonance transfer (the transfer of 
energy from one molecule to another) is made impossible. The energetically rich 
chlorophyll molecules can not transfer their energy surplus to the acceptor 
molecules, but loose this surplus in the form of photones. This is reflected in great 
increasment of fluorescence intensity. 
My goal was to evaluate the efficiency of photosynthetic apparatus in the needles on 
the basis of intensity of primary and secondary fluorescence of chlorophyll. Fresh 
needles without visible external injuries were used for this part of investigation. One 
current-year and one one-year old needle from each tree were choosen for the analy-
sis. The needles were always taken at the distance of 1cm below the twig top or 
below the limit between the current and the one-year old needles, always from the 
upper side of a twig. Cross-sections of the middle part of needles ( 40 x 10-'m thick) 
were done with a criomicrothome (HANOVIA). Autofluorescence of objects in 
water and their secondary fluorescence after addition of DCMU solution was 
observed and photographed. The DCMU solution was prepared by dissolving 0.02g 
DCMU (M=233.1) in 100ml of ethyl alcohol. 
The microscop OL YMPUS - BHS and the photographic equipment of the same 
company were used for the work. The majority of photographs were taken by the 
10x10 magnification, only some details were photographed by the magnification of 
10x20. For the fluorescent microscopy, the exciter filter U was applied (transmits the 
light of the wavelenghts of 334nm and 365nm), the dichroic mirror U (DM-400 + 
L-420) and the supplement barrier filter L-435 and up. The film AGFA CHROME, 
CT 100, DIA was used for ali the photographs. The values on the automatic 
photographic equipment were by ali photographs adjusted to: ASA/400, 
RECIPROCITY/4, EXPOSURE ADJUSTMENT/1. 30% of the eye-field was 
highlighted for each photography. The efficiency of photosynthetic apparatus was 
evaluated on the basis of luminosity of objects. That was also a measure of chloro-
124 
Kmecl A., Batič F., Morfological and anatomical.. 
plast darnage caused by air pollution or other external factors. 
The object on dark background fluoresces red. Its luminosity depends on the size of 
the object (the fluorescing surface) and on its fluorescence intensity. None of the two 
parameters could have been directly measured. 
Size of the object - Each of the objects was photographcd. The objects' contours were 
than copied from the photographs to paper sheets of invariable quality, the shapes 
were cut out and weighed. The size of a whole photograph represented the value of 
100% and the contours of the objects represented corresponding shares of the 
surface. So the relative values of size of the objects were obtained. 
Fluorescence intensity - Because there was no light sensitive cell (photocell) available, 
with which the fluorescence could have been accurately measured, the fluorescence 
was measured with help of the automatic photographic equipment. We assurned, that 
the camera needed equal quantity of light for each photograph. If light was faint, the 
tirne of photography had to be longer. The tirne (t), which was automatically 
adjusted by the photographic equipment, is therefore inversely proportional to the 
object's luminosity (S): 
t = 1/S 
Luminosity is a product of fluorescing surface (P) and fluorescence intensity (IF). 
According to this, the following expression is gotten: 
t = 1/(P x IF) 
For each photograph, the tirne value was registered. The fluorescence intensity was 
then calculated with the following equation: 
IF ;,= 1/(P x t) 
Besides the photographs needed for the analysis of fluorescence intensity in healthy 
needles, some photographs of chlorotic and necrotic needles were also taken. 
5.2 Results 
The results are presented in figures 3 and 4 (the diagrams have been made after the 
values presented in the table - Appendix 2). 
The comparison of autofluorescence intensity and secondary fluorescence intensity 
(after DCMU treatment) in current year needles from different sampling plots is 
presented in the diagram (Figure 3). The course of both curves is contrasting, with 
the assumption, that the extreme values are much more emphasized by the secondary 
fluorescence curve. The greatest difference between the curves occurs for the 
125 
Zbornik gozdarstva in lesarstva, 39 
localities, where the chloroplasts in needles are the least damaged. 
100 --water 
-<)-DCMU 
20 
o 
-"' "\'! -~ -~ 
u .,,; J;l " ~ 
>N E '!: 
'~ " " ~ 
·a-
! 5 .8 E o " l 
::-
e C, "' > ..,, -~ " "' ~ ] " C ~ o e e ~ "" "' ~ v'i "' .s < 5 8 l ;.;l ~ ~ 
N "' >V) '" o ] Q. 
Figure 3: Comparison of autoflorescence intensity and secondary fluorescence 
intensity in current-year needles (spring 1990) 
100 
-<}--DCM 
o 
-"' -"' "' -~ " 
,,;; o 
-~ -E ] ~ >N f.l 
.., 
~ l:! -~ 'i:' ! š 
> P. 
'O ;> "' ·c ~ e e "' "' o lil " " ::. o "' ~ e ii:l i:i -ti < ~ ·e "" ., .s 'O -E ~ N 'O ;> >V) o >O o o.. 
;> o p.. ... 
Figure 4: Comparison of autoflorescence intensity and secondary fluorescence 
intensity in one-year old needles (spring 1990) 
126 
':! ·;:r 
;.;l 
,:,.; 
.. -"' 
:la' .. .... e ~ 
~ o; 
Kmecl A., Batič F., Morfological and anatomicaL 
The comparison of autofluorescence intensity and secondary fluorescence intensity 
(after DCMU treatment) in one year old needles from different sampling plots is 
presented in the diagram (Figure 4). The course of both curves is similar as in 
Figure 3, with identical assumption. In this case as well the greatest difference 
occurs, where the chloroplasts are the least injured and chlorophyll contents the 
highest 
When comparing the curves of autofluorescence of the current-year and one-year old 
needles it can be seen that their courses are similar, but the curve representing the 
current-year needles lies lower than the curve of one-year old needles. That 
indicates, that the chlorophyll quantity in one-year needles is lower than in two-year 
needles. This is normal, and corresponds to our earlier expectations. 
6 DISCUSSION 
Interesting results were obtained with the analysis of chloroplast injuries on the basis 
of chlorophyll fluorescence. It had been expected, that the fluorescence intensity 
depended on the quantity of chlorophyll and integrity of chloroplasts in needles. The 
fluorescence intensity in current-year needles with lower chlorophyll contents (Figure 
3), should therefore be weaker from the fluorescence intensity of one-year old 
needles (Figure 4). 
Despite some deviations and relatively small difference between the fluorescence 
intensities of current-year and one-year old needles it could be said, that the course 
of curves presented in the figures 3 and 4 corresponds to our expectations. Yet 
unusual it is, that our results do not agree with the results of the analysis of 
chlorophyll contents (courtey of Missis Ribarič - Lasnik; Appendix 3). It seems, that 
the mechanism of electron transport has to be taken into consideration. As already 
mentioncd, the DCMU is a herbicide, blocking the photosystem II of photosynthesis. 
That means, that it causes the electron transport breakdown, and this is reflected as 
strong fluorescence increasment. This increasement is the greatest by the objects, by 
which the electron transport before DCMU addition was the least damaged. By the 
objects, where the electron transport had been damaged alrcady before the DCMU 
treatment, the fluorescence increasernent could be almost negligible (a typical 
cxample of this kind is the sampling site Slanica; see Figure. 3). 
With the rnethod tested it is not possible to evaluate the quantity of chlorophyll in 
the needles. The most interesting result, obtained from the curves of 
autofluorescence and secondary fluorescence (after DCMU addition) would certainly 
be their difference. Let us suppose, that DCMU leads to complete (100%) electron 
transport breakdown. If this was trne, the secondary fluorescence would show the 
extent of undamaged electron transport (100% electron transport). Autofluorescence 
represents the actual leve) of damage to elcctron transport rnechanisrn. The 
difference of both values shows, to what degree the mechanism of electrone 
transport is damaged by the examined tree. As the electron transport is of a vita! 
importance for photosynthesis, the results actually directly show the leve! of 
127 
Zbornik gozdarstva in lesarstva, 39 
disturbance in plant's photosynthetic apparatus. Most probably this is one of the best 
presentations of vita! force of a plant which shows, to what degree the plant was able 
to adapt to the environmental changes. 
The tested method should be worked out more precisely. One of the greatest 
deficiencies was, that it was not possible to measure the fluorescence intensity 
directly. Por this purpose, automatic photographic equipment was applied, while for 
completely reliable results highly accurate measuring equipment (photocell) would be 
necessary. Another deficiency could have been, that the needle cuts were too thick 
(40 x l0-6m). Such thickness was necessary, because entire (not torn or otherwise 
damaged) needle cuts were needed for the calculations, but the needles tear very 
easily if the cut is not thick enough. Tearing could probably be prevented by pre-
cooling of the criomicrothome hlade. 
7 LITERATURE 
DENFFER, ZIEGLER 1982: Botanika - morfologija i fiziologija (Botany -
Morphology and Physiology). Škalska knjiga, Zagreb. 
HRČEK D., CIGLAR R., GREGORČIČ B., LEŠNJAK M., PEČENKO A., 
PLANINŠEK T., VEDENIK - NOVAK M., 1988: Proučitev mezoklimatskih 
razmer v občini Velenje (A study of mesoclimatic conditions in Velenje). 
Hidrometeorološki zavod SRS, Ljubljana. 
KALAN, J., in preparation: Vpliv TEŠ na tla in vegetacijo (The influence of Šoštanj 
steam power plant on the soil and vegetation). 2nd part. Inštitut za gozdno in 
lesno gospodarstvo, Ljubljana. 
RIBARIČ - LASNIK, C., in print: Ekofiziološkc lastnosti smreke /Picea abies (L) 
Karsten/ na vplivnem območju Termoelektrarne Šoštanj (Ecophysiological 
characteristics of Norway spruce /Picea abies (L) Karsten/ in the influence 
area of the Šoštanj Steam Power Plant). Magistrsko delo. Ljubljana. 
ŠLIBAR, A., 1990: Morfološke in anatomske spremembe smrekovih iglic (Picea 
abies /L/ Karsten) v vplivnem območju Termoelektrarne Šoštanj 
(Morphological and anatomiceal changes of Norway spruce needles (Picea 
abies /L/ Karsten) in the Šoštanj steam power plant influence area). 
Diplomska naloga. Ljubljana. 
128 
Kmecl A., Batič F., Morfological and anatomical.. 
Appendi.x 1: Sulphur contents (% ); Kalan, 1990 
(l,35 
Wl5 
o 
i -"' -~ "' " -ci -~ "' .C: >N "' 'S! -~ " "' "" ~ -~ '2' o j' " > e ~ p, 1 ., C 'O ,. ·,; 'O o ., ~ 'N' s "' "' ~ "' o C C.:, ;f,J o o "' s v5 N .,. < ti on :.: r;;; "' 3 .; s 'O ~ 
~ :.:: 
N 'O > '"' o ~ ] o 11. p, 
129 
Zbornik gozdarstva in lesarstva, 39 
Appendi.x 2: Fluorescence intensity in current year and one year old needles; 
IF = 1/(P x t). 
Sampling plot Fluorescence intensity 
One year needles Two year needles 
water DCMU water DCMU 
Vrh Smrekovca 11,53 34,87 16,79 73,12 
Koča na Smrekovcu 9,42 88,21 14,44 90,72 
Kramarice 10,62 71,38 13,26 106,44 
Slanica 38,63 48,18 14,87 117,02 
Zavodnje 12,54 63,31 14,91 66,22 
Pod Zavodnjami 13,98 64,06 17,87 53,03 
Lajše 7,87 72,22 14,20 102,80 
Lokovica 15,76 38,99 16,22 51,10 
Veliki vrh 18,17 27,94 13,63 43,79 
Andraž 13,52 77,20 16,23 57,55 
Graška gora 13,55 66,61 16,20 94,07 
Šmiklavž 13,34 54,28 12,02 29,13 
Podgorica 8,81 62,29 12,18 77,42 
Kope 10,54 87,90 12,99 81,02 
Komisija 38,39 76,49 27,19 83,41 
Paški Kozjak 20,16 106,12 16,23 85,32 
130 
Kmecl A., Batič F., Morfological and anatomicaL. 
Appendix 3: Chlorophyll contents spring, 1989 (Ribarič-Lasnik, 1990) 
4 
o.s 
o 
-"' -"' " "' " ,:j " -~ 
..C >N \'! '~ "' " "' -"' i:: ~ u <.> '2 !? ?~ > "' " o. :;r "' 't:: '2 "O -o o "' ·;:: o 'N' s s "' "' ~ " ..J o C q ~ o ~ e o (/,) s ;;; N -"' eJ> ~ l,'.l "' 'i) <( o E "O o 
-E " ~ N -o j > ,v:, o ~ o.: '" o o.. > lil o. 
Explanation to the photographs 1 and 2 
Both photographs show a cross-section of a Norway spruce needles by fluorescence 
rnicroscopy, 
On Photograph 1, the autofluorescence of chlorophyll in needle cells can be scen 
and on J:lhotograph 2 the seconda1y fluorescence of the same object after addition of 
fluorochrorne DCMU ( dichloro-phenyl dime til urea). 
The difference between fluorescence intensities shows to which leve! is damaged the 
photosynthetis system of the needlc;, under examination, The lowest the difference, 
the more chlorophyll has been damaged, or in other words, the smaller is photosyn-
thetis activity of the needle and of the whole tree. 
This rnethod of evaluation shows, to which degrce is damaged the photosynthetic 
rnechanism of a tree and thus directly illustrates the leve] of tree's vitality. 
The method is used on camparative basis. The photographs were taken on OL YM-
PUS BHS microscope, with photographyc equipment of the same producer (10 x 10 
magnification, fluorescence michroseopy), 
131 
Zbornik gozdarstva in lesa rstva, 39 
Photographs l and 2: 
132