Published September, 1980
NOTES
age capacity. We simply have the computer store only the
number of root intersects in a vertical scan and then dump
the rest of the data for that scan, thus drastically reducing
storage requirements. Rotating the photographic image 90°
did not affect the goodness of fit, thus indicating the validity
of using nonrandom vertical scans. This may not hold true
for all types of root systems, but for corn and soybeans, we
have encountered sufficient root branching to prevent a biased
orientation of root placement during photography.
This method of determining root length is fast. Table 1
shows the time required to complete three scanning replications.
Included in this time interval is the calculation of root length
plus hard copy printout of each of the three replications and
their average. A scanning interval of 14 columns (35 vertical
scans per replication) provides accurate and fast sampling.
Routine Root Scanning Procedure
Once the photography technique has been developed for a
given root system (image with proper contrast between root
and background), a set of calibration pictures is taken using
known lengths of string or thread with size and color similar
to the roots, keeping all conditions constant (i.e., camera focal
length, size of root photography area, lighting conditions, film
type, and development). When root pictures are to be digitized,
the calibration pictures are digitized first. The computer then
calculates the calibration regression equation and stores it for
later conversion of root intersects into root length. Root trans-
parencies can be mounted in individual slides or left uncut in a
continuous strip.
The scanning and digitizing takes only 5 sec per picture.
Eye fatigue is minimal using this technique. One person can
easily digitize over 500 root pictures in 8 hours under ideal
operating conditions that require a minimum of adjustment
on the digitizer. This compares with about 25 pictures a day
for one person using older versions of Newman's technique.
Roots can be scanned directly, rather than using photographic
images. We use the photographic technique for three reasons:
1) it generally is not feasible to have the computer and digitiz-
ing equipment in the same laboratory where the roots are
being prepared; 2) too much computer "dead time" would be
wasted waiting for the next root sample to be prepared and
positioned prior to scanning; and 3) the photographic tech-
nique provides a hard copy, permanent record of the roots.
Original cost of the system (excluding computer) in 1974 was
~$15,000. While this cost is substantial, it does represent a
tremendous savings as compared with the manual labor and
time needed to determine root length by other line-intersects
methods. This equipment can also be used for other measure-
ments, like leaf area determinations and other image processing
procedures.
851
A SCREEN-CAGED THERMOCOUPLE
PSYCHROMETER AND CALIBRATION
CHAMBER FOR MEASUREMENTS OF PLANT
AND SOIL WATER POTENTIAL
Ray W. Brown and James M. Collins1
ABSTRACT
Estimates of water potential with thermocouple psy-
chrometers are subject to errors due to temperature gradi-
ents. The objective here is to describe the design and
construction procedures of a new screen-cage single-junc-
tion psychrometer that minimize these errors. The screen-
cage design restricts entry of contaminants into the psy-
chrometer cavity; allows rapid vapor equilibration be-
tween the sample and the psychrometer; and provides a
360° concentric evaporation surface that reduces the in-
fluence of vapor condensation due to temperature gradi-
ents. In addition, a calibration chamber that can also
be used for plant and soil samples is described.
Additional index words. Thermocouple psychrometer,
Temperature gradient effects, Water potential estimates,
Psychrometer design criteria, Screen-caged psychrometers.
ECENT evidence shows that temperature gradi-
R ents cause spurious estimates of water potential
with thermocouple psychrometers (15, 16). Errors due
to temperature gradients are particularly severe with
double-junction units and with those that are enclosed
within end-window type chambers. Double-junction
psychrometers (5, 8) mask the detection of tempera-
ture gradients by reducing the magnitude of zero-offset
voltages to near zero, hence errors are not readily
evident (16). Psychrometers with end-window geome-
try enclosed in solid chambers of either Teflon (4) or
stainless steel can trap condensed water within the
chamber. Voltage outputs from these units can be
either erroneously low or high as a result of tempera-
ture gradients (15). Psychrometers with both the dou-
ble-junction and end-window design features are most
subject to error.
Single-junction thermocouple psychrometers with a
concentrically located junction surrounded by a cy-
lindrical sample surface appear to be least subject
to temperature gradient errors (15). Ceramic units,
such as those manufactured by Wescor (Wescor, Inc.,
Logan, UT 84321)2 and screen-caged units (2) gen-
erally meet these criteria; however, both models have
some advantages. Ceramic cup units lag behind actual
changes in soil water potential, particularly under
drier conditions when liquid contact is lost between
soil particles and the ceramic (2). Also, salt adsorp-
tion by the ceramic matrix may be a problem in saline
soils. The original screen-caged psychrometers (2)
were designed with a large-mesh screen that may have
allowed soil particles or other matter to contaminate
the thermocouple junction (6), although recent re-
search showed this to be insignificant (4). In addition,
1
Plant physiologist and forestry technician for the Intermoun-
tain For. and Range Exp. Stn., Forest Service, USDA, Ogden,
Utah 84401, respectively, stationed at the Forestry Sciences Lab.,
Logan, Utah. Received 5 July 1979.
"Use of trade or firm names is for reader information only,
and does not constitute endorsement by the USDA Forest Ser-
vice of any commercial product or service.
 a52 AGRONOMY JOURNAL, VOL. 72, SEPTEMBER-OCTOBER 1980

the original screen-caged model was difficult to cali-

brate and did not provide for measurements of am- E PVC INSULATED CABLE

bient temperature.
Increased use of thermocouple psychrometers to .MELTABLE,SHRIINK
TUBING LINER
measure soil and plant water potentials in both the
field and the laboratory requires a more reliable de- POLYPROPYLENE’ SHRINK
TUBING LINER
sign. Also, it would be advantageous to measure the
MYLAR SHIELD
water potential of in situ soil, soil samples, or plant &COLOR-CODED INSULATION
tissues interchangeably. We present design criteria for
a new screen-caged psychrometer that appears more re- EPOXY RES IN
liable and versatile, together with design features for
a unique calibration chamber to be used with this
psychrometer. -TEFLON PLUG

U
Design and Construction

T h e single-junction screen-caged thermocouple psychrometer

is illustrated in Fig. 1. T h e cable (Revere Corp. of America,
I’ ’7
1; COPPER-CONSTANTAN JUNC TlON
(soldered)

Wallingford, CN 06492)* is custom constructed, and consists of il \ I I!

two copper leads and one constantan lead, each 26 AWG and
separately insulated with color-coded PVC (polyvinyl chloride).
T h e three leads are wrapped with a n aluminized mylar shield
and a 26 AWG bare Cu drain wire. T h e cable is coated with a n
C H R cm)
(0.0025
WELDED
O M E L - v
II I
I/’ CONSTANTAN
(0.0025 cml

outer layer of PVC insulation. JUNCTl ON .I 11,400 MESH INNER LINER

T h e stripped ends of the Cu and constantan lead wires are II 002 . 11 MESH OUTER LINER
inserted through a prepunched Teflon plug (0.47 cm diam) II
and allowed to extend about 2 cm out the other end. One of
the protruding Cu leads (designated as electrically negative) is
bent over 90” and wrapped once around the constantan wire.
IIII li
!I
Unlike previous designs, this Cu-constantan junction provides
for measurements of ambient temperature. T h e remaining Cu
lead (positive) and the constantan wire are trimmed to a length -------STAINLESS STEEL SCREER CAP
of about 2 mm beyond the Teflon plug surface. I.Ocm
T h e chromel-constantan (Omega Engineering Co., Stamford, I

CN 06907)* welded thermocouple is soldered to the extruding Scale

Cu and constantan lead wires using stainless steel solder and
flux. Our thermocouples are constructed by similar methods to
those described by Lopushinsky (7), except that a Cu electrode
is used instead of graphite. T h e chrome1 wire of the thermo-
couple is attached to the positive Cu lead and the constantan
wire to the constantan lead. T h e thermocouple is attached so
that the junction can be positioned 5 mm beyond the Teflon
plug, a point corresponding to the center of the psychrometer
cavity suggested by Wiebe et al. (16). T h e thermocouple is
first carefully soldered onto the protruding lead wires, then
the trailing ends of the thermocouple wires are wrapped se-
curely around the lead wires, and resoldered. These solder
joints are now the reference junctions of the psychrometer. We
suggest the above procedure be performed under a dissecting
microscope using ultrafine precision forceps to facilitate manip-
ulation of small components.
T h e stainless steel screen cage is constructed of a n outer
cylinder of 200 mesh and a n inner cylinder of 400 mesh wire
cloth. T h e outer cylinder is custom constructed (Wayne Wire
Cloth Products, Inc., Kalkaska, MI 49649)a of type 304 stainless
steel 200 mesh wire cloth, 1.3-cm in length by 0.48-cm inside
diam (ID) with a welded seam. One end of the outer cylinder
is sealed with a 0.48-cm-diam Teflon disk covered with a 200
mesh stainless steel screen disk soldered in place. T h e inner
cylinder is constructed of 400 mesh stainless steel screen that
has been rolled into shape and inserted into the outer cylinder.
T h e inner cylinder is held in place by expansion pressure
against the wall of the outer cylinder. T h e completed screen
cages should be thoroughly washed in a sonic cleaner to remove
all flux and foreign matter. Then, the cage is pushed over
the Teflon plug; care must be taken not to disturb the exposed
thermocouple. It is essential that the screen cage fit snugly
over the Teflon and that it be positioned so the sensing junc-
tion is located halfway between the two Teflon surfaces.
A small amount of epoxy resin cement is applied to the back
side of the Teflon plug, over the lead wires, to prevent move-
ment and possible electrical shorting. Before the cement dries,
a double layer of shrinkable tubing (Penntube Plastics Co.,
Clifton Heights, PA 19018)2 is positioned over the portion of
the screen cage that encloses the Teflon plug and u p into the
cable PVC insulation. Both layers (1.3-cm in length) are heated
Fig. 1. Illustration of the single-junction screen-cage tht:rmo-
couple psychrometer (drawn to scale).
simultaneously with a hot air blower to about 150 C. The
inner layer of shrinkable tubing is a meltable liner of poly-
propylene, which, when heated, flows into the screen pores and
adheres to the cable insulation. T h e outer layer provides a
firm protective shield that is permanently bonded to the melt-
able liner. Together the two layers provide a watertight seal
that adds rigidity to the psychrometer.
Psychrometers of this design are presently available from
J. R. D. Merrill Specialty Equipment (Logan, U T 84321) and
EMCO (P.O. BOX 34, Angola, IN 46703).2
Calibration Chamber
Calibration of screen-caged psychrometers is performed by
suspending them over known molalities of NaCl solutions in
sealed chambers under isothermal conditions (2, 16). We
found the best chamber for this purpose to be commercially
available stainless steel tube fittings (Fig. 2) consisting of both
a reducer and a cap (either Swagelok SS-400-R-10 and SS-1010-C,
respectively, Crawford Fitting Co., Solon, OH 44139; or Gyro-
lok 4R10-316 and 1OCP-316, respectively. Process Instruments
and Control, Salt Lake City, U T 84115).2 T h e two pieces of
the fitting should be slightly modified, as illustrated in Fig. 2,
according to the following specifications: 1) the orifice of the
reducer should be bored to a diam of 0.64-cm to allow free
passage of the psychrometer, and; 2) the rim above the th.reads
of both the reducer and the cap should be machined to a
smooth surface with a 15” angle to provide a seat for “0” ring
seals. T w o double “0” ring seals are required, one in the reducer
orifice and another in the lower cap (PAI-1-010, 0.25 in I:D by
0.375 in outside diam (OD), and PAI-1-114, 0.625 in ID by 0.813
in OD, respectively, (Precision Assoc., Inc., Minneapolis, MN
55401).a Also, a stainless steel spacer (0.95-cm OD, 0.64-cm. ID,
and 0.32 cm-thick) is required to compress the “0” rings over
the reducer. T h e seal around the cable is provided by a -piece
of surgical tubing (2.5-cm in length, 0.64-cm OD by 0.32-cGt ID)
behind the psychrometer.
 NOTES 853
Of some concern is the possibility that contaminants may
PSYCHROMETER CABLE enter the psychrometer cavity through the screen by liquid
flow in saturated soils (6, 9). We have not observed such con-
tamination even after repeated long-term exposures to saturated
soils under field and greenhouse conditions. Recalibration after
UPPER CAP
these tests showed no significant drift. Liquid water will not
penetrate the 400 mesh screen liner except under pressure.
SPACER These units have been completely immersed to a depth of 20
0" RINGS cm in NaCl solutions, then briskly agitated, and yet after
15 "ANGLE (modification) periods of up to a week (the maximum period tested) no solu-
tion penetrated the screen cage. After this extreme treatment
SURGICAL TUBING the psychrometers still produced expected microvoltage outputs
REDUCER for that solution. Apparently the pocket of trapped air within
REDUCER ORIFICE (modification) the screen cage is not easily displaced, even in liquid media.
The screen-caged psychrometer appears to meet all the cri-
teria suggested by Wiebe and Brown (14) and by Wiebe et al.
0" RINGS (15) to minimize errors caused by ubiquitous temperature gradi-
ents (12) under field conditions. In addition, the screen cage
15 ° ANGLE (modification) permits more rapid equilibration with evaporating surfaces
than other materials, such as ceramic, because it does not im-
SCREEN CAGE, PSYCHROMETER
pose a restrictive barrier to water vapor movement.
Screen-caged psychrometers should be thoroughly cleaned be-
LOWER CAP fore use. We found that new units can be contaminated with
SAMPLE CHAMBER solder flux, oil, or grease within the screen cage. Tuner cleaner
in pressurized cans (e.g., LPS Part No. 00720, LPS Research
Lab., Los Angeles, CA 90025),3 commonly used to clean electrical
components, is an excellent means of removing contaminants
by liberally spraying directly through the screen cage. This is
Fig. 2. Illustration of the stainless steel calibration chamber followed by a vigorous wash with liberal quantities of distilled
water administered through the nozzle of a squeeze bottle. Par-
showing seals and screen-cage thermocouple psychrometer tial drying is then achieved with filtered pressurized air, fol-
(drawn to scale). lowed by total drying in an oven at 80 C for several hours or
air-drying overnight. If an oven is used, it is essential that the
temperature not be allowed to exceed 90 C, otherwise severe
Calibration and Application damage to the PVC insulation on the cable will result.
The calibration chamber can be used for many applications
other than calibration, such as measurements of water potential ACKNOWLEDGMENT
in plant tissues, soil samples, or other media. The psychrometer The authors appreciate helpful suggestions during the prepara-
is first sealed within the reducer (Fig. 2), but with the lower tion of the manuscript by Jed Merrill, Dr. W. T. McDonough,
cap unattached. For calibration, a filter paper liner (What- and Dr. H. H. Wiebe.
man No. 1, 25-cm X 4-cm) is rolled onto the end of a stainless
steel tube (0.64-cm ID X 0.95-cm OD, and 15-cm in length)
and inserted into the chamber around the psychrometer. The
liner is held in place around the tube with the finger of one
hand, but expands against the inside walls of the chamber
when released. The tube can then be extracted, leaving the
filter paper behind surrounding the psychrometer and lining
the entire cavity of the chamber. Calibration solution can then
be applied directly to the filter paper liner with either a
syringe or from a drop-bottle. When completely saturated, the
excess should be poured off, and the cap replaced immediately
to avoid evaporation. The threaded portion of the cap should
be loosened slightly to allow the "0" rings to easily slide over
the reducer and to allow equilibration of atmospheric pressure.
After sliding the cap completely over the reducer, the threads
can be finger tightened to effect the seal.
Plant leaf tissue can be similarly sealed within the chamber
surrounding the psychrometer. The chamber is particularly well-
suited for the insertion of grass leaves using the stainless steel
tube, but many broad-leaved species such as aspen (Populus
tremuloides Michx.) have also been used successfully. The spa-
cious geometry of the chamber permits use of large leaf samples
with the advantage of low cut edge-to-volume ratios (1, 10, 11,
13). At 25 C in isothermal water bath conditions, we found
that about 2 hours of equilibration time is sufficient for both
calibration solutions and most leaf tissue. Nonrepresentative
tissue water potentials due to metabolic changes may result
with much longer equilibration times (11).
During the last 2 years, we have tested approximately 200
screen-caged psychrometers measuring in situ soil water po-
tential in the field, greenhouse, and laboratory. Until recently,
we had 135 units in the field, each horizontally positioned to
lessen temperature gradient effects (9). These psychrometers
had been monitored periodically for over a year (3), during
which time they all continued to provide both soil temperature
and soil water potential data as expected. Twenty-seven of them
were extracted in the spring of 1978 and laboratory tested by
methods similar to those of Brown and Johnston (4). Recalibra-
tion showed that a 1-year exposure to field conditions caused
only a 2.9% drift from original calibration data.
854 AGRONOMY JOURNAL, VOL. 72, SEPTEMBER-OCTOBER 1980
ABSORPTION AND TRANSLOCATION OF
CHROMIUM THROUGH THE SURFACE
OF A SOYBEAN LEAF»
D. J. Pickrell and B. G. Ellis9
ABSTRACT
Although Cr is known to be important in animal meta-
bolism, it does not readily translocate through the roots of
most plants. The addition of Cr to plant leaves as a spray
material is a possible method of increasing the Cr con-
tent of plants. The objective of this laboratory study
was to determine if Cr would be absorbed by a plant
leaf and translocated to other portions of the plant. A
single drop containing radioactive B1Cr or raFe (for com-
parison) as ethylenediamine tetraacetic acid (EDTA)
was placed over the mid-rib of one leaf on a soybean
[Glycine max (L.) Merr.] plant. Absorption and trans-
location of the tagged Cr or Fe were followed by radio-
autographs while plants were growing and by counting
separated plant parts at the end of the experiment. Both
El
Cr and MFe were absorbed through the surface of the
soybean leaves and translocated throughout the plant,
but only small amounts were translocated (4% of absorbed
Cr). The absorption and translocation of Cr closely
paralleled that of Fe. The results show that soybean
plants will absorb Cr through their leaves and translocate
it to other portions of the plant but the rate for these
processes is relatively slow.
Additional index words: Glycine max (L.) Merr., nCr,
ra
Fe, Cr absorption and translocation.
HROMIUM (Cr) has been the subject of much
C recent interest because of its importance in animal
metabolic systems and its possible role as the active
ingredient in glucose tolerance factor (GTF) (Mertz,
1975). Slight Cr deficiency in laboratory rats is charac-
terized by a mild diabetic state (Schroeder, 1967) and
has been linked to an increase in circulating choles-
terol, degeneration of the cornea by vascular infiltra-
tions and opacities (National Academy of Sciences,
1 manner. Autoradiograms were made by exposing the intact
Contribution of the Crop and Soil Science Dep., Michigan
Agric. Exp. Stn. Journal Article No. 9096. Received 20 July
1979.
'Former NSF High School Honors science student and pro-
fessor, respectively, Crop -and Soil Sci. Dep., Michigan State
Univ., East Lansing, M1 48824. The Senior author wishes to
acknowledge the NSF scholarship for the High School Honors
Science Program under which this work was accomplished.
1974) and a possible increase in aortic plaques (Schroe-
der, 1967). Laboratory exclusion of Cr from rat diets
led to retarded growth and diabetes mellitus, reversible
by addition of trace amounts of Cr3+ acetate to drink-
ing water.
The effect of Cr on human health may be compar-
able. Mertz (1975) found an improved glucose toler-
ance in certain adult Americans after dietary supple-
mentation of CrCla, and Schroeder (unpublished re-
port) showed that the rate of diabetes in a country
increases in direct proportion to the use of refined
sugars. This increase is suspected to be caused by the
removal of trace minerals including Cr in the refine-
ment process. Symptoms of excessive dietary intake of
Cr in man are unknown (National Academy of Sci-
ences, 1974) although deficiency of Cr in human tissues
is quite prevalent, especially in the United States
(Schroeder, 1976).
The ideal way to reduce tissue Cr deficiency would
be to increase Cr concentration in the foods we eat.
However, this has proven to be unexpectedly difficult.
Schueneman (1974) found that as little as 50 ppm. Cr
applied to sandy soils produced wilting in tomatoes
[Lycopersicum esculentum Mill.] and crop growth
was reduced up to 70% compared to controls for to-
matoes and corn [Zea mays L.]. Alloway (1968) re-
ported that plants suffering from induced Cr toxicity
frequently contained the same percent of Cr in leaves
and stems as found in unaffected plants. Gary et al.
(1976) mentioned that plants which did accumulate
Cr from nutrient solutions retained most of this Cr
in their roots. They also found that those plants which
do accumulate Cr are those that will accumulate: Fe
as well. In many respects, the chemistry and behavior
of Cr3* and Fe3"1" compounds are analogous, yet the
ease o£ reduction of 3
the Fe2+3+ — Fe2+ couple as com-
pared with the Cr * — Cr couple may account for
greater mobility of Fe in plants (Gary et al., 1976>).
Although increasing the Cr content of a plant does
not mean that it becomes useful to animals or humans,
it is the first step in improving Cr nutrition.
The purpose of this experiment was to determine if
Cr could be absorbed and translocated after applica-
tion to the surface of the leaves. Since the behavior
of Cr and Fe is often analogous, and since much infor-
mation exists on absorption of Fe by leaves, Fe was used
for comparison.
MATERIALS AND METHODS
Soybean [Glycine max (L.) Merr.] seeds (Cultivar Hark) were
germinated in acid-washed quartz sand with distilled, deionized
water. One week after seeding, individual plants were removed
from the sand; their roots were washed with distilled deionized
water; and the plants were transferred to plastic vessels con-
taining 2 liters of continuouly aerated Hoagland's solution.
Radioactive Cr, as ^CrEDTA was applied to a soybean leaf by
placing a single drop over the mid-rib of the center trifoliate
leaf of a 2-week-old soybean plant. For comparison purposes,
another plant was tagged with raFeEDTA solution in a similar
leaves to X-ray film for 2 hours at intervals of 4, 24 and 48
hours after tracer application. After the exposure at 48 hours,
the leaves were washed with dilute detergent and distilled
water. Additional exposures were then made: 2 hours a.t 48
hours after tracer application, and 30 hours at 72 hours after
tracer application. After the last autoradiograms (102 hours
after tracer application), each plant was divided into the ter-
minal bud including the latest trifoliate leaf, upper leaves.