11. Ramirez, J. M. 1977. Influence of data of harvest on yields and 15. Smoot, J. J. and P. W. Hale. 1977.

. 1977. Evaluation of decay control

leaf carbohydrates of citrus. PhD Dissertation, Univ. of Florida,
Gainesville.
12. Rivero, L. G. 1978. Distortion studies of ‘Marsh’ grapefruit. MS
Thesis, Univ. of Florida, Gainesville.
13. Sarig, Y. and D. Nahir. 1973. Deformation characteristics of ‘Va-
lencia’ oranges as an indicator of firmness. HortScience 8:391-392.
1 4 . ---------------- and S. Orlovski. 1974. Viscoelastic properties of
‘shamouti’ orange./. Text. Studies 5:339-349.
treatments and shipping containers for export of grapefruit to
Japan. Proc. Fla. State Hort. Soc. 90:152-154.
16. Soule J., W. Grierson, and J. G. Blair. 1967. Quality tests for citrus
fruit. Fla. Agr. Expt. Sta. Cir. 315.
17. Vines, H. M., W. Grierson, and G. J. Edwards. 1968. Respiration in-
ternal atmosphere, and ethylene evolution of citrus fruit. Proc. Amer
Soc. Hort. Sci. 92:227-234.

/. Amer. Soc. Hort. Sci. 104(4):554-557. 1979.

Anthocyanins of Fruits of Vaccinium, Sub-genera

Cyanococcus and Polycodium 1
Walter E. Ballinger, G. J. Galletta, and E. P. Maness
D epartm ent o f H orticultural Science, N orth Carolina State University, Raleigh, N C 27650
A dditio n a l index words, taxonomy, chemotaxonomy
Abstract. Anthocyanins of ripe fruits of 13 species of Vaccinium (12 species of the Cyanococcus and 1 species
of the P olycodium sub-genera) collected in the eastern United States and grown together near Castle Hayne,
North Carolina were hydrolyzed to yield aglycones and sugars. TLC separation and identification of these hydrol-
ysates indicated that anthocyanins of the 12 species of the sub-genus Cyanococcus (true blueberries; Camp)
contained 5 aglycones (> means “in greater quantity than’’) (delphinidin > cyanidin > malvidin > petunidin >
peonidin) and 2 sugars (galactose > arabinose). Anthocyanins of Polycodium representative V. stam ineum (deer-
berries) differed from those of representatives of Cyanococcus in that they yielded only large amounts of the
aglycone cyanidin and trace amounts of peonidin. Hydrolysis of anthocyanins of fruits of V. stam ineum and 7
species of Cyanococcus yielded glucose. Thus, species of Cyanococcus appeared to have a similar anthocyanin
content. Anthocyanin contents of P olycodium (V. stam ineum ) were different than those of Cyanococcus but
generally the same as those reported for cranberries (V. macrocarpon, sub-genus O xycoccus). Reports in the
literature indicate that anthocyanins of other sub-genera of Vaccinium contain xylose (V. m yrtillis; bilberry;
cyanidin-3-xylosylglycoside) and rhamnose (V. vitis-idaea; cowberry; delphinidin-3-glycoside, 5-glycoside, and
3-rhamno-glycoside). Thus, as indicated from this initial study, fruits of the genus Vaccinium appear to vary in
anthocyanin content with sub-generic classification.

Harborne (7) considered the pattern of the anthocyanin
contents of ericaceous plants to be very consistent and the
berries of plants of species of genus Vaccinium in the Ericaceae
family to have a complex range of anthocyanins. Timberlake
and Bridle (8) later wrote that arabinosides and galactosides
frequently occur in the Ericaceae. Francis, Harborne, and Barker
(4) reported that one of the extensively-grown fruit crops in
Northeastern United States, lowbush blueberries (V. angusti-
folium), has one of the most complex anthocyanin contents
of any plant. They extracted and identified 15 anthocyanins
from these fruits. These yielded 5 of the 6 commonly occurring
aglycones (delphinidin, petunidin, malvidin, peonidin, and
cyanidin) and 3 sugars (3-monoglycosides) (arabinose, galac-
tose, and glucose). Ballinger, Maness, and Kushman (1) later
found a similar content of anthocyanins in highbush blueberries
(V. corymbosum). Cranberries (V. macrocarpon) reportedly
contain 6 anthocyanins (cyanidin-3-glucoside, peonidin-3-
galacto side, cy anidin-3 -arabinoside, cyanidin-3 -galactoside,
peonidin-3-arabinoside and peonidin-3-glucoside (5).
From a chemotaxonomic point of view, the genus Vaccinium
appears to be characterized by the general presence in it’s fruits
iReceived for publication May 10, 1978. Paper No. 5514 of the Journal
Series of the North Carolina Agricultural Experiment Station, Raleigh,
NC 27650.
The cost of publishing this paper was defrayed in part by the payment of
page charges. Under postal regulations, this paper must therefore be
hereby marked advertisement solely to indicate this fact.
^USDA New Crops Branch exploration grant (SR-9).
of some or all of the various 3-monoglycosidic combinations of
the aglycones delphinidin, petunidin, malvidin, peonidin, and
cyanidin with the sugars galactose, arabinose, and gluocse.
Therefore, a study was conducted to test the hypothesis of
Harborne concerning the consistently complex anthocyanin
profiles of Vaccinium on the basis of a new and richer col-
lection of germplasm.
Methods and Materials
Clonal and seed samples of 30 Vaccinium species reported
growing in the eastern United States were collected by Dr. G.
J. Galletta.2 These were planted in a field at Castle Hayne, N.C.
This is a “middle latitude” of their natural distribution, a geo-
graphical location where most of the plants from New England
to Florida could survive and grow. When the planting was old
enough to permit a uniform, representative sampling of fruits,
12 species of the sub-genus Cyanococcus, including 2 species
originally collected from more than one state and one species of
the sub-genus Polycodium (Table 1), were selected. These
species were considered to have potential for broadening the
genetic base in commercial blueberry improvement.
Each week in 1972 from June 5 to August 12, ripe fruits of
5 seedlings each of the above species, plus 3 bushes each of 4
cultivars of V. corymbosum/V. australe and V. ashei included
in the planting as “internal standards,” were sampled when the
fruits of a given bush had reached their peak of ripening. The
berries were returned to Raleigh, where the number of fruit
in 50-g of ripe fruit of each sample was counted before the
samples were frozen. Later, the anthocyanins were extracted
quantitatively in 250-ml or less of 1% HC1 in methanol. Extracts

554 J. Amer. Soc. Hort. Sci. 104(4):554-557. 1979.

Table 1. Sugars hydrolyzed from anthocyanins of fruit of species of 2 sub-genera of Vaccinium.
Sugar (visual rating)2
Fruit wt Total ACY
(g/berry) (mg/ 1 0 0 -g) Arb Gal Glu^
Species & sample Ploidy Fruit
Sub-genus source (state) no. color Mean SD Mean SD Mean SD Mean SD Mean SD

A. Wild species representatives

Polycodium staminium (Georgia) 2 light 0.54 0.13 161 63 1.4 0.5 2.4 0.5 U.O u.u
Cyanococcus ashei (1) (Florida) 6 med 0.52 0 .1 0 521 82 2.2 0.5 3.0 0.0 0.2 0.5
ashei (hybr.) (Georgia) 6 med 0.52 0 .1 0 501 89 2.2 0.5 3.0 0.0 0.2 0.5
ashei (2) (Florida) 6 med 0.48 0.04 556 146 2.0 0.0 3.0 0.0 0 .0 0.0
amoenum (hybr.) (Georgia) 6 dark 0.52 0.09 675 138 1.0 0.9 3.0 0.0 1.2 1.1
amoenum (Arkansas) 6 dark 0.40 0.06 570 79 2.4 0.9 2.8 0.5 1.2 0.8
corymbosum (New Jersey) 4 med 0.51 0.14 503 123 2.4 0.5 2.8 0.5 1.6 0.5
australe (North Carolina) 4 med 0.48 0.05 638 74 2.4 0.5 3.0 0.0 0 .0 0. 0
australe (Florida) 4 med 0.40 0.03 454 81 2.2 0.5 3.0 0.0 0.0 0.0
australe (New Jersey) 4 med 0.37 0.09 506 109 2.8 0.5 3.0 0.0 0.2 0.5
australe (hybr.) (North (Carolina) 4 med 0.35 0.08 623 73 1.8 0.5 2.8 0.5 1.2 1.1
myrsinites (Florida) 4 dark 0.30 0 .0 2 522 12 2.0 0.0 2.8 0.5 0.0 0.0
angustifolium (Maine) 4 light 0.28 0.06 295 63 1.0 1.0 1.0 1.0 3.2 0.5
pallidum (Arkansas) 4 light 0.33 0.11 273 110 2.0 0.0 2.3 0.6 1.7 0.6
tenellum (North Carolina) 2 dark 0.29 _ X
454 - 3.0 3.0 0.0
darrowi (Florida) 2 light 0.25 0.09 355 183 1.6 0.9 3.0 0.0 0 .0 0.0
myrtilloides (Maine) 2 light 0.19 0.05 254 54 0.8 0.5 0.3 0.5 3.0 0.0
atrococcum (Georgia) 2 dark 0.17 0.04 563 133 2.8 0.5 3.0 0.0 0.0 0.0
elliottii (Florida) 2 med 0.11 0.03 764 107 2.6 0.5 3.0 0.0 0 .0 0.0

B. Cultivars (improved)
‘HomebelT ashei (Georgia) 6 dark 1.22 _ X
470 - 3.0 - 3.0 - 0. 0 -
‘Tifblue’ ashei (Georgia) 6 light 1.35 _ X 272 - 2.0 - 3.0 - 1.0 -
‘Croatan’ corymbosum (North Carolina)w 4 med 0.89 _ X
276 - 3.0 - 3.0 - 0 .0 -
‘Bluecrop’ corymbosum (New Jersey)w 4 light 1.56 _ x 140 - 3.0 - 1.0 - 1.0 -

zGal = galactose; Arb = arabinose; Glu = glucose; SD = standard deviation (5 seedlings at random/species); the higher the rating, the larger the relative
size of the spot on the TLC plates for a given species.
yRatings of 0.0 for Glu do not necessarily indicate the complete absence of Glu on the anthocyanins of fruit of a given species, rather, that Glu was
not detectable in the quantities of hydrolysate used for the determinations. This was especially true for species whose fruit had anthocyanins with
small amounts of Glu.
xOne sample only, therefore no SD.
wMost improved highbush blueberry cultivars are given the original Latin name assigned by Linnaeus, V. corymbosum, even though they are species
hybrids that are preponderantly derived from V. australe. Small with somegermplasm from V. corymbosum L. and V. augustifolium aiton.

were brought to volume with methanol and HC1 to yield a pH
meter reading of 1.0. Total anthocyanin content on a 100-g of
fruit basis was calculated using the optical density of the ex-
tracts determined spectrophotometrically at 535 nm (1).
Subsequently, the extracts were concentrated and purified
using paper chromatography (1). Ten-mg of the purified and
dried anthocyanins per sample was hybrolyzed; sugars and
aglycones were separated and later identified using thin layer
chromatography (TLC) ( 1 ) . For this, 20 x 20 cm, microcrystal-
line cellulose plates (Schleicher and Schuell, Inc.) were scored
to provide 20 1-cm wide channels. Two concentrations of each
composite sample of aglycones plus standard cyanidin were
spotted in duplicate on a plate which was then developed
first in formic:HC1:H20 (10:1:3; by volume) and then in
amyl:HAc:H20 (2:1:1; by volume) (1). The plates were dried.
The proportions of separated aglycones were rated visually from
1 to 5 (the larger the spot, the higher the score). The mixture of
sugars hydrolyzed from each sample plus standards of glucose,
galactose, and arabinose were spotted on 4 plates. Two of
these plates were developed with the solvent phenol:H20
(4:1; by volume) (1). The other 2 were developed with butanol:
benzene:pyridine:H20 (5:1:3:3; by volume) (1). The developed,
dried plates were sprayed with 2.5% aniline hydrogen phthalate
for visualization of sugar spots and visually rated for relative
distribution of sugars (1). Since quantities of aglycones and
sugars on the plates were rated visually (subjectively) over a
period of several weeks, a quantitative comparison of ratings
across species would not be valid.
Data for the 5 samples per species were used to calculate
means and standard deviations of berry weight, total antho-
cyanin content, and types of aglycones and sugars resulting
from the hydrolysis of the anthocyanins.
Results
Berry weight (Table 1). Weight of berries of the sub-genus
Cyanococcus generally followed published ploidy number for
the species (Camp, 1945) with the hexaploids associated with
some of the largest-sized fruits and the diploids associated with
some of the smallest-sized fruits. Vaccinium stamineum fruits
were as large as any of the unselected seedlings in the test.
Total anthocyanin content (Table 1). All samples of the
sub-genus Cyanococcus seemed to have as much or more total
anthocyanin content as ‘Bluecrop’ fruit. Cultivars selected for
light blue color ( ‘Bluecrop’ and ‘Tifblue’) and species known
for light colored fruit (V. stamineum, angustifolium, myrtil-
loids, darrowi, and pallidum) generally had less anthocyanin
content (average for this group was 250 mg/100-g) than dark
fruited clones such as ‘HomebelT and species like V. amoenum,
myrsinites, tenellum and atrococcum (group average 542 mg/
100-g). Interestingly, the species and clones which are normally
considered a medium blue in color showed as much anthocyanin
(534 mg/g average) as the dark fruited taxa. However, it is
common knowledge that color of blueberries is influenced as
much by the amount of waxy bloom on their surface as by
anthocyanin content.
Sugars hydrolyzed from anthocyanins (Table 1). Without

J. Amer. Soc. Hort. Sci. 104(4):554-557. 1979. 555

Table 2. Aglycone content (mean and standard deviation) of the acid hydrolysates of anthocyanins of fruit species of two sub-genera of Vaccinium.

Aglycone (visual rating) 2

Species & source P lo id y ______ __________ ^ y _______ Mv_______ P t______ Pn
Sub-genus (state) no. Mean SD Mean SD Mean SD Mean SD Mean SD
A. Wild species representatives
Polycodium staminium (Georgia) 2 0.0 0.0 4.0 0.0 0.0 0.0 0.0 0.0 0.1 0.0
Cyanococcus ashei ( 1 ) (Florida) 6 3.8 0.5 3.8 0.5 2.2 0.5 1.8 0.9 1.0 0.0
ashei (hybr.) (Georgia) 6 3.6 0.5 3.6 0.9 2.4 0.9 1.8 0.5 0.8 0.4
ashei (2) (Florida) 6 2.8 0.5 4.0 0.0 2.2 0.5 1.6 0.5 1.0 0.0
amoenum (hybr.) (Georgia) 6 3.2 0.7 3.2 1.0 2.8 0.7 1.7 0.5 1.5 0.5
amoenum (Arkansas) 6 2.8 0.5 3.4 0.9 2.0 0.0 1.8 0.5 1.0 0.0
corymbosum (New Jersey) 4 3.8 0.5 F8 0.9 2.8 0.9 2.4 0.5 0.5 0.5
australe (North Carolina) 4 3.8 0.5 F8 0.5 2.8 0.5 2.8 0.5 0.8 0.4
australe (Florida) 4 4.0 0.0 3.0 0.7 2.2 0.5 1.8 0.5 0.1 0.0
australe (New Jersey) 4 4.0 0.0 2.8 0.9 2.2 0.9 2.0 0.0 0.5 0.5
australe (hybr.) (North Carolina) 4 3.8 0.5 3.4 0.5 1.4 0.5 2.0 0.0 0.6 0.5
myrsinites (Florida) 4 3.6 0.5 2.4 0.9 2.6 0.5 2.2 0.9 0.6 0.5
angustifolium (Maine) 4 3.0 0.7 2.8 0.5 3.0 1.0 2.0 0.0 1.0 0 .0
pallidum (Arkansas) 4 3.0 0.0 2.0 0.0 2.7 0.6 1.7 0.6 1.0 0.0
tenellum (North Carolina) 2 4.0 _y 1.0 - 2.0 — 2.0 — 0.1 —

darrowi (Florida) 2 3.0 0.7 2.4 1.1 1.2 0.8 1.6 0.5 1.2 0.8
myrtilloides (Maine) 2 2.7 0.5 3.0 1.1 3.0 0.0 1.0 0.0 0.8 0.5
atrococcum (Georgia) 2 3.8 0.5 1.6 0.5 2.8 0.5 2.6 0.5 0.1 0.0
elliottii (Florida) 2 3.6 0.5 2.8 0.9 2.0 0.0 1.8 0.5 0.5 0.5

B. Cultivars (improved)
‘HomebelF ashei (Georgia) 6 4.0 - 3.0 - 2.0 - 2.0 - 1.0 -

‘Tifblue’ ashei (Georgia) 6 2.0 - 4.0 - 1.0 - 1.0 - 1.0 -

‘Croatan’ corymbosum (North Carolina) 4 4.0 - 3.0 - 2.0 - 4.0 - 0.1 -

‘Bluecrop’ corymbosum (New Jersey) 4 4.0 - 2.0 - 3.0 - 4.0 - 0.1 -

zDp = delphinidin; Cy = cyanidin; Mv = malvidin; Pt = petunidin; Pn = peonidin; SD = standard deviation (5 seedlings at random/species).
yOne sample only, therefore no SD.

exception, the anthocyanins of all samples of fruit yielded
arabinose and galactose; 7 of the 12 Cyanococcus species
yielded detectable amounts of glucose. Fruits of 2 species,
V. myrtilloids and V. angustifolium, had anthocyanins with
unusually large quantities of glucose, much more than arabinose
or galactose. Geographic origin of the plants of V. ashei and
V. australe did not appear to influence sugar content of the
anthocyanins. Sugars other than the three above were not
detected.
Aglycones (Table 2). The anthocyanins of fruit of V stam-
ineum (sub-genus Poly codiurn) had a much different content
of agly cones than that of the other spp. (sub-genus Cyanococ-
cus). Vaccinium stamineum was associated with large amounts
of cyanidin, a small amount of peonidin, but no delphinidin,
malvidin, or petunidin. Anthocyanins of other species yielded
5 algycones. No aglycones other than the 5 aglycones above
were detected in any of the samples. Overall, relative quantities
of aglycones detected from anthocyanins of fruit of the sub-
genus Cyanococcus appeared to be in the order, delphinidin >
cyanidin > malvidin > petunidin > peonidin. Aglycones hy-
drolyzed from the anthocyanins of the cultivars included in the
planting as internal standards (V. ashei and V. australe) were
generally the same as those of the fruits of the sub-genus Cyano-
coccus plants obtained from the wild. The geographic origin
of the spp. V. ashei and V. australe did not appear to influence
the aglycone contents of the anthocyanins of their fruits.
Discussion
Fruits of all species of Cyanococcus studied thus far contain
anthocyanins which, upon hydrolysis, yield the 5 aglycones,
delphinidin, petunidin, malvidin, cyanidin, and peonidin, plus
the 2 sugars, arabinose and galactose; glucose is often present
(1, 4). Vaccinium myrtilloides and V. angustifolium, since
their anthocyanins yielded glucose in quantities obviously
greater than those of other species of the sug-genus Cyanococcus,
should perhaps be studied further to determine if they have a
phylogenetic relationship. The data offered herein did not shed
any light on the putative origins of the polyploid Vaccinum spp.
deduced by Camp (3) on the basis of morphological, distribu-
tional, and chromosome level finding [see Galletta (6) for a
summary and updated treatment of Cyanococcus].
Polycodium representative V. stamineum appears to differ
from Cyanococcus representatives in anthocyanin content.
Of the 5 aglycones hydrolyzed from anthocyanins of Cyano-
coccus, only cyanidin was present in appreciable amounts in
anthocyanins of V. stamineum. Vaccinium stamineum antho-
cyanins had more galactose than arabinose which was true also
for anthocyanins of Cyanococcus. Anthocyanins of V. stam-
ineum appear to resemble those of cranberries (V. macro-
carpon, sub-genus Oxycoccus (5) more than they do the antho-
cyanins of Cyanococcus.
Anthocyanins of fruits of V. vitis-idaea and V. myrtillis
appear different from those of all 3 sub-genera above. The
anthocyanin of V. myrtillis (bilberry - Euvaccinium sub-genus)
has been reported to be cyanidin-3-xylosylglycoside while the
anthocyanins of V. vitis-idaea (ling or cowberry — Vitis-idaea
sub-genus) reportedly are delphinidin-3-glycoside,-5-glycoside,
and -3-rhamnoglycoside (9). Upon acid hydrolysis of the type
used in our present study, anthocyanin extracts of these 2 fruits
would have yielded the sugars xylose (cowberry) and rhamnose
(bilberry). Our study did not reveal the presence of any sugars
other than arabinose, galactose, and glucose.
Thus, fruits of the genus Vaccinium do not appear to have as
uniform and consistent an anthocyanin content as hypothesized.
Differences in anthocyanins appear to be associated with the
sub-generic classification proposed by Camp (3).

556 J. Amer. Soc. Hort. Sci. 104(4):554—557. 1979.

 Literature Cited
1. Ballinger, W. E., E. P. Maness, and L. J. Kushman. 1970. Antho-
cyanins in ripe fruit of the highbush blueberry. J. Amer. Soc. Hort.
Sci. 95:283-285.
2. ___________ , ____________ , G. J. Galletta, and L. J. Kushman.
1972. Anthocyanins of ripe fruit of a “pink-fruited” hybrid of
highbush blueberries. J. Amer. Soc. Hort. Sci. 97:381-384.
3. Camp, W. H. 1945. The North American blueberries with notes on
other groups of Vacciniaceae. Brittonia 5:203-275.
4. Francis, F. J., J. B. Harborne, and W. G. Barker. 1966. Anthocyanins
in the lowbush blueberry, Vaccinium angustifolium. J. Food Sci.
31:583-587.
5. Fuleki, T. and F. J. Francis. 1967. The co-occurrence of monogluco-
sides and monogalactosides of cyanidin and peonidin in the Ameri-
can cranberry, Vaccinium macrocarpom. Phytochemistry 6:1705-
1708.
6. Galletta, G. J. 1975. Blueberries and Cranberries, p. 154-196. In
J. Janick and J. N. Moore (eds), Advances in fruit breeding, Purdue
Univ. Press, West Lafayette, Ind.
7. Harborne, J. B. 1967. Comparative Biochemistry o f the Flavonoids.
Academic Press, New York.
8. Timberlake, C. F. and P. Bridle. 1975. The anthocyanins. Chap. 5
of The Flavonoids, ed. by J. B. Harborne, T. J. Mabry, and H. Mabry.
Academic Press, New York.
9. Troyan, A. V. and I. F. Borukh. 1968. Biologically active substances
of wild-growing berries of the Carpathians. Tr. Vses. Semin. Biol.
Aktiv. (Lech.). Veshchestvam Plodov. Yagod., 3rd. [Chem. Abstr.
73, 119394; 1968.]

J. Amer. Soc. Hort. Sci. 104(4):557—560. 1979.

Penetration of Photosynthetically Active Radiation as a

Measurement of Canopy Density of Citrus Trees1
Otto L. Jahn2
Agricultural Research , Science and Education A dm inistration , U.S. D epartm ent o f Agriculture,
Orlando , FL 32803
Additional index words, defoliation, leaf area, Citrus spp.
Abstract. Citrus trees were defoliated to obtain measurements of radiation penetration at various canopy densi-
ties. Tree size measurements were used to calculate the leaf-area index (LAI) and leaf-area to canopy-area ratio
(LAC) of trees. Penetration of photosynthetically active radiation (PAR) increased curvilinearly as defoliation
increased and LAC decreased. Observed relationships between PAR penetration and LAC were used to estimate
LAC values from PAR penetration measurements. Correlations between PAR penetration and LAC were slightly
higher than with LAI. LAI and LAC appeared to vary independently from tree size as measured by tree diameter
or leaf area.

Heinicke (3, 4) showed that light penetration in apple trees
was nonlinear, with a change in leaf density affecting light
penetration more at low than at high leaf densities. More
recently, measurements of light penetration have been used to
estimate the LAI of apple trees (1). In citrus a dense canopy
develops at the outer periphery of the tree, resulting in little
penetration of light beyond the first 1 m of canopy depth (2).
The density of this canopy is indicated by counts of 24,446
leaves on a 6-year-old tree and 142,728 on a 15-year-old orange
tree. These trees had estimated leaf areas of 74 and 296 m2,
respectively (Selhime, personal communication). Turrell (9)
obtained leaf counts of 16, 37, 93, and 173 thousand on 3-,
6-, 12- and 29-year-old orange trees, respectively. Leaf areas
were reported as 34, 59, 146 and 203 m2. Formulas were de-
veloped for estimating leaf area from tree age and trunk diam.
In preharvest degreening studies with ethephon, defoliation
losses of more than 10% may occur (10). Recent work showed
that most of these losses were of older, inside leaves (5). Al-
though losses of over 20% were found, no consistent effect on
PAR penetration was shown and there was no reduction in
yield the following year. These observations prompted further
studies on methods of measuring canopy density and its rela-
tion to PAR penetration with the objective of developing a
rapid method of estimating leaf density.
1Received for publication November 3, 1978.
The cost of publishing this paper was defrayed in part by the payment of
page charges. Under postal regulations, this paper must therefore be
hereby marked advertisement solely to indicate this fact.
^Research Horticulturist.
Materials and Methods
Trees were selected for uniformity of canopy density and for
a range in size. A total of 16 trees of 7 cultivars were studied
in 1975, 1976, and 1977. Repeated ethephon applications were
used to defoliate selected trees. Fallen leaves were collected,
air-dried, and weighed at frequent observation dates. Separate
samples were counted, measured with an area meter, and dried.
Calculated leaf area or number per gram dry weight from these
samples was then used to estimate the number and area of
leaves for each tree and date of collection. For dry weight
determinations, leaves were oven-dried at 70°C. In 1976 and
1977, estimates of defoliation were obtained by counting leaves
on 8 to 20 twigs per tree at each observation date.
Radiation penetration measurements were taken on each
tree initially and at each leaf collection date. Photosynthetically
active radiation (PAR) was measured as juE m"2s_1 on a meter
with a quantum sensor (400-700 nm sensitivity). The sensor
was held 25 cm above and aimed at the center of a 0.4-m2
reflectance board painted white. In use, the board was placed
horizontally at preselected points on the ground around the
tree trunk. In 1975, 4 positions per tree were used. In 1976,
additional measurements were made on the north side of 3
of the preselected points. In 1977, an additional 4 points
were added about 1 m inside the edge of the canopy. Measure-
ments of direct sunlight and unshaded reflected PAR were also
recorded.
Light measurement time varied from tree to tree, but mea-
surements for each tree were made at the same time for all
observation dates. All trees were measured within an hour of
12 noon. No attempt was made to adjust for-the changing
sun angle occurring over the 4- to 6-week duration of a test.

J. Amer. Soc. Hort. Sci. 104(4):557-560. 1979. 557