TOBACCO,
A SENSITIVE MONITOR
FOR  PHOTOCHEMICAL
AIR  POLLUTION
  U.S. DEPARTMENT OF HEALTH, EDUCATION, AND WELFARE
            Public Heollh Service
   Consumer Protection and Environmental Health Service

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 TOBACCO, A SENSITIVE MONITOR
                    FOR
 PHOTOCHEMICAL AIR POLLUTION
                      by
                  Walter W. Heck
                  Frank L. Fox
                 C. Stafford Brandt
                      and
                  John A. Dunning
                Agricultural Section
          Division of Economic Effects Research
U. S. DEPARTMENT OF HEALTH, EDUCATION, AND WELFARE
               Public Health Service
    Consumer Protection and Environmental Health Service
       National Air Pollution Control Administration
                 Cincinnati, Ohio
                    June 1969

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Dr.  Walter W. Heck and Dr. C.  Stafford Brandt are employees of the U.S.
Department of Agriculture,  but are  assigned to the Division of Economic
Effects Research of the  National  Air Pollution  Control Administration.
The AP series  of reports is issued by the National Air Pollution Control
Administration to report the results of scientific and engineering studies,
and  information of general  interest in the field of air pollution.  Information
reported in this series includes  coverage of NAPCA intramural activities
and  of cooperative studies conducted in conjunction with state and local
agencies,  research institutes, and industrial organizations.  Copies of AP
reports may be obtained upon request,  as supplies permit, from the Office
of Technical Information and Publications,  National Air Pollution Control
Administration, U. S.  Department of Health,  Education, and Welfare,
Ballston Center Tower No.  2,  801 North Randolph Street,  Arlington,
Virginia  22203.
     National Air Pollution  Control Administration Publication No.  AP-bs

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                            CONTENTS
ABSTRACT	   iv



INTRODUCTION	      1



PILOT STUDY I	        	      	    3



PILOT STUDY II	      	        	    7



    Soil and Nutrient Combination-1   	      	    7



    Soil and Nutrient Combination-2   ...    	    8



    Growth Interactions	      9



    Shading Interactions    	     .        	    .   10



    First Summer Monitoring Program     	   10



    Second Summer Monitoring Program	    	     11



DISCUSSION	   15



APPENDIX.  TOBACCO MONITORING PROGRAM PROCEDURE   ...   17



REFERENCES	23
                                    iii

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                              ABSTRACT

      The development of a technique by which the sensitive tobacco variety,
Bel W3 is used as a monitor for photochemical air pollution is discussed.
The technique uses the plant as an indicator of the  oxidant complex in both
urban and rural areas.  Two pilot studies that were conducted over a 3-year
period during the development of the monitoring technique are included in the
discussion.   Attention is given to an explanation of the proper procedures for
planting, transplanting, fertilizing, and caring for mature  plants.  The
methods used in determining and recording injury to plant leaves is included;
the studies showed almost daily  injury to monitoring plants.
KEY WORDS:  tobacco, monitor,  photochemical,  oxidant, plant,  and air
               pollution
                                     iv

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       TOBACCO,  A SENSITIVE MONITOR
                                FOR
       PHOTOCHEMICAL  AIR POLLUTION

                         INTRODUCTION
     Indigenous vegetation has been used to identify the photochemical com-
plex, to help map the extent of photochemical pollution,  and to provide some
evidence in support of control programs.  The use of natural vegetation as a
pollution monitor,  however, is limited by variations in sensitivity because of
differences in naturally occurring cultural and environmental  conditions, non-
specificity in injury symptoms because of poor care of plants, and lack of
uniform distribution of species within a. monitoring area.

     Various attempts have been made to assess naturally occurring pollu-
tion loads and to identify specific "smog"  components by special planting of
a specific variety of plant under more or less controlled conditions.    The
most comprehensive program of this type was attempted in Los Angeles with
annual bluegrass and petunia as the monitoring species. ^  in a similar pro-
gram, pinto bean plants were used. 3  The monitoring plants were grown under
controlled conditions in both studies and exposed to ambient conditions for a
24-hour period. Attempts to correlate injury with oxidant level were only
partially successful.  MacDowall et al. 4 correlate sensitivity  of tobacco with
oxidant level by using an empirical factor (the coefficient of evaporation).
This factor was developed from several meteorological factors and has been
used with some success in forecasting oxidant injury to tobacco.

     At present no technique that combines  ease  of use  and relatively uniform
sensitivity of plant material over a long period of time has been developed for
use by an inexperienced operator.  The development of such a technique
employing Bel W3 tobacco is discussed herein.

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                            PILOT STUDY  I
      Air pollution problems  that relate to greenhouse management prompted
a study to determine the utility of tobacco as  a monitor of ambient oxidant.
Tobacco is  excellent for this  type of study for several reasons:  It has been
more intensively studied than any other photochemically sensitive species;
it has an  indeterminate growth habit during vegetative stages; mature leaves
are most sensitive and show fairly  uniform sensitivity at a given stage of
growth; new injury is readily separated from old injury; it can be grown under
uniform cultural  conditions with a minimum of care; a highly susceptible
variety, Bel W3, has been developed;^ its symptoms  of oxidant injury are
characteristic, easily identifiable,  and quite  specific.  Figures  1 and 2 show
characteristic oxidant (ozone) fleck on Bel W3 tobacco.
  Figure 1. Character! sti c oxidant (ozone) injury to
         Bel W3 tobacco variety showing complete
         destruction of older leaves.
Figure 2- Single tobacco leaf shows 20 to 25 percent
       Injury from oxidant (ozone) fleck.
      A pilot study was conducted in the metropolitan areas  of Boston,  Cin-
cinnati,  and St.  Louis during the summer months to determine whether Bel
W3 tobacco could be used as a simple  field monitor and to answer three basic
questions:

       1.  Are oxidant concentrations in metropolitan areas, which presum-
          ably are from photochemical sources, sufficient to produce mark-
          ings on the leaves of sensitive vegetation?

       2.  Can estimates of the frequency of injury be obtained?

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       3.  Can a quantitative estimate of injury be related to concentration or
          dose of oxidant?

      Fifty-five greenhouse operators  in the three metropolitan areas agreed
to participate in  the study.   Each grower was asked to situate two tobacco
plants inside his  greenhouse.  He was  to provide at  least a bushel of his
normal potting soil, water  the plants as needed,  and follow his standard dis-
ease control and  fertilizer  practices.  Each grower was instructed in the
identification of the symptoms of oxidant injury to the tobacco and was
requested to examine the plants  daily and record new injury by both leaf and
plant number.  Cincinnati locations were inspected  every 2 weeks by a
National Air Pollution Control Administration staff  member, and detailed per-
centage injury indices were obtained.

      Detailed data obtained from the growers in the three metropolitan areas
cannot be reported because of the variability in cultural conditions and  care
of plants and because of the irregularity and doubtful reliability of observa-
tions by the greenhouse  personnel.   Thus,  that the sensitivity of exposed
material throughout the  area was the same  or that results noted were uni-
formly recorded  is not known.  As a result, no estimates  can be made  in
regard to the regional distribution of any one episode of oxidant injury nor
can estimates of differences in severity among episodes be made.  From the
data obtained, however,  some conclusions  can be drawn.  On some occasions
during the season, oxidant concentrations were sufficiently high to cause
markings on sensitive vegetation.  All locations were subject to episodes of
elevated oxidant  concentration sufficient to mark  sensitive vegetation.  There
appeared to be some correlation between the occurrence of high oxidant con-
centrations at the Continuous Air Monitoring Program Stations and reports
of tobacco injury within the next 3  days in both Boston and St. Louis.   Some
of the growers correlated appearance of tobacco  injury with appearance of
similar injury patterns  on  other greenhouse crops.   However,  the use  of
observers and different cultural conditions  in this type of study is of  only
limited value.

      The sensitive variety of tobacco used in the study  offers  communities
a method of detecting  the presence of phytotoxic  concentrations of oxidant
and provides a method for determining the frequency of  occurrence and gen-
eral estimates of regional distribution and level of oxidant.   However,  greater
control over cultural conditions, exposure, and collection of data  must be
exercised.

      Results of  the biweekly inspection of the Cincinnati plants are summa-
rized for the period from July 7 through September  9.  Injury as a. percentage
value for each leaf was determined and reported  on  the basis of total plant
injury (average of two plants).  Possible environmental  factors that affect
sensitivity were  obtained and are included with the injury indices in Table 1.
Results suggest that evaporative cooling has had  the greatest effect on  sensi-
tizing plants to ambient  oxidant.  This correlates with lower maximum
greenhouse temperatures and is probably a moisture stress effect.  No evi-
dence of correlations between sensitivity and fertilizer schedule,  and distance
and  direction from town was found.  However, correlation of shade,  soil
texture, type of house, and care of plants was noted.  Thus  growth and expo-
sure conditions are important to such a. study. Results  suggest no significant
difference in oxidant concentrations  within an 8-  to  14-mile  radius of the
center of Cincinnati.
                                               TOBACCO, A SENSITIVE MONITOR

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          Table I.  INTERACTION OF FACTORS ON  RESPONSE  OF TOBACCO

              TO OXIDANT POLLUTION IN METROPOLITAN  CINCINNATI
Environmental factors
Average injury index
Temperature
Evaporative cooling 95°F
Fan only 105°F
No cool ing 1 15°F
Soil texture
Light
Medium
Heavy
Type of house
Florist
Vegetable
Care of planets
Good
Med i um
Poor
Fertilizer schedule
None
1-8 appl ications
10 or more apol ications
Shade, percent
0-10
15-45
above 50
Distance from center of town
7 mi les or less
7 to 13-1/2 miles
Direction from town
W-NW
N-NE
SE
Number of
locations
2k

6
2
16

8
9
7

16
8

11
8
9

8
12
4

10
10
4

14
10

11
12
1
Injury index3
545 (0-1820)

1200
565
300

645
630
320

700
230

895
425
175

410
625
590

365
630
790

520
580

595
475
840
         Values  are  reported as percent cumulative leaf injury  per  plant.
         Each  plant  value  is obtained by adding the individual  injury
         percentages from each leaf.
      Factors affecting tobacco sensitivity were determined from the study
and separated into controllable and non-controllable variables (Table 2).
These factors were considered when setting up the second study.
Pilot Study I

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       Table 2.  FACTORS AFFECTING THE SENSITIVITY OF TOBACCO

                  TO PHOTOCHEMICAL AIR POLLUTANTS3
     Factors
                              Controllable
                                                      Not controllable
Light intensity
Soil  structure and
  compaction


Nutrient level
Soi1  moisture



Insects and diseases


Light duration


Light quali ty


Temperature



Hum i d i ty


Wind  speed


Ambient pollutants
Use a 50 percent shade
cloth so plants are not
exposed to full sunlight

Use a light, friable mix
with high water-holding
capabi1i ties

Use regular fertilizer
with high nitrogen
content

Water daily, the 1ight
soil mix will drain well
and give good aeration

Minimize and schedule
use of controls
Partial reduction of
leaf temperature using
shade cloth
Partial reduction with
shade cloth
 Natural
variations
 Natural
variations
 Natural
variations

 Natural
variations

 Natural
variations
 Natural
variations

 Natural
variat ions

 Natural
variations
 These factors were determined from the first pilot study.  They
 affect sensitivity during the growth of the plant as well as during
 the exposure period.
                                                     TOBACCO, A SENSITIVE MONITOR

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                          PILOT  STUDY II

      Results of the first study enabled researchers to develop a second study.
Its procedure,  modified following its completion,  appears in the Appendix.
The second study was designed to answer four  questions:

       1.  What is  the best soil mix and nutrient combination to use for maxi-
          mum plant sensitivity and good growth?

       2.  How long is the tobacco sensitive and what growth habit is
          preferable?

       3.  What shade factor should be used?

       4.  What is  the area of influence of the photochemical complex in the
          Cincinnati area?

SOIL  AND  NUTRIENT COMBINATION-1
      Three  soil  types (peat-perlite,  peat-perlite-soil, and peat-perlite-
s oil-manure) and four nutrients (Hoagland, Plant Marvel, * Plant Marvel and
Cal-Mag, and tap  water) were used in this experimental design.  Each nutrient
except tap water was used with each soil type.  Tap water was used only with
the peat-perlite-soil-manure soil to give ten soil-nutrient combinations.
Four  replicates were used per treatment combination,  and the complete set of
ten soil-nutrient combinations was tested seven times  during the summer
period, for a total of 28 replications per treatment.

      Tobacco was seeded,  transplanted after 4 weeks into the different soil
types in 4-inch plastic pots, and grown for 1 week with the appropriate nutri-
ent in a greenhouse in which the air was  charcoal filtered.  The plants were
then placed outside under a 50 percent shade cloth, and exposed to the ambient
air for 2 weeks.  A new experimental replicate was started each week to allow
an overlap.  Injury indices  were recorded every day for 2 weeks, then the
plants were discarded.  Indices for average injury per leaf for each plant
were  obtained and used to determine results  shown in  Table 3.  The plants
grown with tap water showed poor  growth with  obvious nutrient defiency;
and although they were the most sensitive, their poor  growth made them poor
monitors for air pollution.  Plants grown in the Hoagland nutrient were more
sensitive than those fertilized with Plant Marvel and Plant Marvel plus Cal-
Mag but did not grow as well.  Considering health of plants, sensitivity, and
work  involved in care and maintenance of monitoring sites, peat-perlite-soil
with a 20-20-20 nutrient addition  (this could be the Plant  Marvel)  is recom-
mended for use with the monitoring plants.
*Mention of company or product name does not constitute endorsement by the
Department of Health, Education, and Welfare.

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           Table 3.  INTERACTION OF SOIL  AND  NUTRITION ON RESPONSE
                     OF TOBACCO TO AMBIENT  OXIDANT LEVELSa
Nutrient
Soil Hoagland Plant Marvel
Plant Marvel
Cal-Mag
Tap
Water
Average
Soi 1
Peat-perl i te
(1/1)
Peat-perl i te-
soi 1
(1.5/1.5/1.0)
Peat-perl i te-
soi 1-manure
41
43
43

31
37
37

35
34
35

I 36
38
(54)b 39

 Average  nutrients
                     42
35
               35
  Values  are  reported as average injury per  leaf on a percentage basis for
  those  leaves showing injury from ambient  pollution.  All values are
  averages of 28 replications.   Mean values  connected by  lines are not
  statistically different.  Summary shows no  difference  between the soils,
  although there appears to be an interaction  between nutrient and soil in
  the  case of Plant Marvel and peat-perlite.   The Plant Marvel and Plant
  Marvel  plus Cal-Mag are not different from each other,  but are different
  from the other nutrients.  Hoagland and tap  water are different in the
  peat-perlite-soi1-manure mix.

  The  value listed for tap water was not  included in the  average for the
  peat-perlite-soil-manure mixture.

SOIL AND  NUTRIENT COMBINATION-2

      Two soil types (peat-perlite-soil and peanut hulls-soil) and two nutrients
(Kapco and Plant Marvel plus  Cal-Mag) were used in this experimental design.
Kapco was used only with the peat-perlite-soil; the second nutrient was used
with both soils.   The tobacco was grown with initial transplant as detailed in
the Appendix.  The second transplant  into  bushel baskets was into the experi-
mental soil type with the appropriate nutrient.  Each treatment was replicated
12 times in a. randomized block design and  was an integral part of the study on
growth effects.  Plants were exposed  to the ambient air under a 50 percent
shade cloth.

      Injury  indices were obtained  daily (Monday through Friday), and cumula-
tive injury indices for each plant were reported after 4, 8,  12,  and 15 weeks.
Mean values  (12 replications)  for each treatment are reported in Table 4. No
evidence of a nutrient effect was noted.  After the  first 4-week period signifi-
cantly less injury to plants grown in the peanut hulls-soil mixture than to
those grown  in the peat-perlite-soil mixture was noted.   This was probably
due to the longer transplant recovery  period in the heavier peanut hull-soil
mixture.  No other 4-week period showed  significant differences,  although
the last 3-week period showed greater injury to plants  in the heavy mix.
This  may have been due to the greater amount of uninjured tissue on the
plants in the  heavier soil treatment prior to the last 3-week period.  Seasonal
values for the three treatments are the same.  The heavier soil is  not recom-
mended for monitoring purposes,  because severe wilting at transplant results

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in a slower start.  The peat-perlite-soil mixture with a 20-20-20 nutrient
addition is recommended.

        Table  k.   EFFECT OF FERTILIZER AND  SOIL  TYPE  ON SENSITIVITY
                   OF BEL W3 TOBACCO TO AMBIENT POLLUTIONS
Soi 1/ferti 1 izer
Peat-perl i te-soi 1
(1.5/1.5/1.0)
Plant Marvel
20-20-20
Alternate Cal-Mag
Peat-perl i te-soi 1
(1.5/1.5/1.0)
Kapco 20-20-20
Peanut hul 1 s-soi 1
Plant Marvel , 20-20-20
Alternate, Cal-Mag
Cumulative injury indices
7/13
Ct week)
915
870
695b

8/10
(8 week)
1255 (2170)
1285 (2155)
1275 (1970)

9/7
(12 week)
830 (3000)
905 (3060)
865 (2835)

9/28
(15 week)
555 (3555)
515 (3575)
670 (3505)

 Values are reported as percent cumulative  leaf  injury per plant.   Each  plant
 value is obtained  by adding the individual  injury percentages from each leaf.
 Value is statistically different from the  other  two values listed on 7/13  at
 the 0.01 level.

GROWTH  INTERACTIONS
      Tobacco was chosen as a monitoring plant in part because it produces
new leaf tissue throughout most of the growing season.  In conjunction with
the nutrient study,  three  growth habits were studied for each soil-nutrient
combination for a total of 12 replicates per growth habit.  These  were grouped
in studying the growth effects, because the  soil-nutrient had no effect.  One
group of plants grew normally throughout the season producing,  after 8 to 10
weeks,  tall,  rather unwieldy plants that were difficult to handle.  A second
group was topped after 8  weeks and the regrowth was studied.  A third  group
made use of new plants every 4 or 8 weeks.  With 12 plants per  nutrient
treatment, each growth treatment contained four plants.  In the three growth
treatments (one per soil-nutrient combination) in which new plants  were used,
the four  plants per treatment were started together.  Two of  the four
plants  in one soil-nutrient combination were replaced with young  plants after
2 weeks.  Two of the four plants in each of the other two soil-nutrient com-
binations were replaced by young plants after 4 weeks.  Thereafter,  the two
older plants in the  first soil-nutrient combination were  replaced  every  2
weeks  and the two older plants in each of the other two combinations  were
replaced with young plants every 4 weeks .

      Cumulative injury indices  were obtained as in the soil-nutrient study
and are reported in Table 5.  Because it takes a. week to 10 days  for newly
transplanted plants to show maximum sensitivity, the cumulative indices for
the replacement plant series were obtained from only two plants  for the 10
days after plants were  replaced.  Values were not different after the first
10 days.  Plants were topped after 8  weeks, and results showed that it took
about 4 weeks for these plants to regain sensitivity.  Plants grown during
Pilot Study

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full season and replacement series plants were no different after 12 weeks.
Plants transplanted during September developed slowly and did not produce
as much sensitive tissue.  Thus,  they show less injury during the last 3-
week period.  The regrowth was well started a'fter 4 weeks and contained con-
siderable sensitive tissue during the last 3 weeks.  Plants grown full season
were in flower,  and all leaves, including the floral bracts,  showed  injury at
the end of the 15-week exposure period.

        Table 5.   EFFECT OF GROWTH PERIODS  ON TOTAL INJURY TO BEL W3
                      TOBACCO FROM AMBIENT POLLUTIONS


Growth period
Ful 1 season
4- and 8- week
growth alterna-
ting replacement
every 2 and 4
weeks
Topping after
8 weeks wi th
regrowth
Cumulative injury indices^
7/13
(4 week)
815
840




820


8/10
(8 week)
1285 (2100)
1235 (2075)




1300 (2120)


9/7
(12 week)
915 (3015)
820 (2895)




**190 (2310)


9/28
(15 week)
660 (3675)
--425 (3320)




670 (2980)**


 aValues are reported  as  percent cumulative leaf  injury per plant.   Each plant
  value  is obtained by adding the individual  injury percentages from each  leaf.

  Values with asterisks are statistically different from the other  two  values
  listed for the same  date at the 0.01  level.

      Plants that were allowed to develop for a full season were the most
 severely injured;  however,  the  injury was not much greater than that seen in
 the replacement series.  Because  of difficulties inherent in working with
 large plants, a  replacement sequence of 6 weeks -alternating replacement
 of two of the four  plants every 3 weeks   is recommended in the procedural
 outline in the Appendix.

 SHADING  INTERACTIONS
      Four  plants were grown as detailed in the Appendix on an 8  to 4 replace-
 ment basis  with final transplant into bushel baskets under 0,  35, 50, and 80
 percent shade.  Injury indices "were obtained and reported as in the earlier
 studies.  Results  are given as cumulative average plant values for the paired
 plants in the different shade treatments (Table 6).  Results suggest that plant
 sensitivity is reduced if plants are left in the sun during the mid-summer
 months, possibly because of higher temperatures,  light intensity,  and drought
 conditions,  all of which may cause soil moisture stress.  A 50 percent shade
 cloth is recommended for all monitoring work to protect  the plants.

 FIRST SUMMER MONITORING PROGRAM

      Plants were grown as detailed in the Appendix on a 4-week alternating
 replacement basis at seven locations in the vicinity of Cincinnati,  Ohio.
 Cumulative leaf injury on a percent basis for each plant was obtained on a
 weekly basis at each location.  Continuous oxidant readings were  recorded at
 10
                                                TOBACCO, A SENSITIVE MONITOR

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         Table 6.   EFFECT  OF  SHADE ON SENSITIVITY OF TOBACCO  BEL W3
                        TO AMBIENT OX IDANT LEVELS3
Date
replaced
7/06/66
8/03/66
8/31/66
9/28/66
Length of
exposure,
weeks
it
8
8
8
Plants
(pai red)
odd
even
odd
even
Cumulative injury indices,
percent shade
0
1)90
1500
1658
1313
35


1673
1280
50
615
1902
1553
1295
80
608
2070
1680
1293
  Values are reported as  percent cumulative leaf injury per  plant.  Each plant
  value is obtained by adding  the  individual injury percentages  from each leaf.
  Definite reductions are noted with the 7/6 and 8/3 comparisons  between 0
  shade and both the 50 and  80 percent shades.   These are somewhat  tenuous
  because only two plants were used for each data point.
the laboratory (Mast) and corrected to 2 percent neutral KI values.  The
weekly oxidant index was recorded  as the cummulation of hourly averages
that exceeded 3. 0 parts  per hundred million (pphm) of oxidant during the hours
between 6 a.m.  and 10 p.m.  All hourly averages from the time the oxidant
level reached 3. 0 pphm were included through the last one above 3. 0,  even
if some of the mid values fell below 3. 0.  The total was then arbitrarily
divided by 2 to obtain the final index.  Plants were lost  at two locations  from
high winds.   Although the recorded oxidant  injury -was similar to injury  to
plants at the other five locations, data from the two locations are not included
in the summary table.   Table 7 includes the weekly oxidant index, the weekly
plant injury index for the major study (Tables  4 and 5),  and the  weekly plant
injury index for five of the  monitoring sites.  Distance and direction from the
center of Cincinnati are included  as well as the season's total cumulative
indices.  The injury index to  oxidant index ratio was determined for the two
groups of plants located at  the oxidant monitoring site.

     No consistent  relationship between oxidant values and plant injury was
found.  Meteorological factors were not included in this study.   Possibly an
inclusion of these would produce a correlation between oxidant and injury.
The levels  of sulfur dioxide and nitrogen dioxide would be of interest in  view
of the reported synergism between  sulfur dioxide  and both ozone and nitrogen
dioxide.  The data suggest  that within the limits of the study there is little
difference in the phytotoxic potential of the  pollution complex at any location
monitored.   There is a.  suggestion that  at certain times one station has  a
higher phytotoxic potential  or that meteorological conditions are such that
plants show variable sensitivity at the five locations.  Injury to the two  groups
of plants at the laboratory was remarkably  uniform.

SECOND SUMMER MONITORING PROGRAM
     Plants were grown as detailed in  the Appendix on a 3-week alternating
replacement basis at five locations east of Cincinnati, Ohio. Stations were
located at 5, 7,  25,  50, and 75 miles east of the center of the city.  Injury
indices and oxidant indices were generated  as outlined, and results are shown
in Table 8.
Pilot Study II
                                                                          11

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                                      Table 7.   RESULTS  OF  FIRST  CINCINNATI AREA  SUMMER MONITORING PROGRAM3

Location
Oxidant
(laboratory)13
Laboratory - 1
(Tables 4 and
5)
Laboratory - 2
(Mon i tor s i te)
Dunning
Matthis
Clark
Ross
Distance (miles)
and d i rect i on
from Cincinnati
5(E)

5(E)


5(E)

18(E-NE)
25(E)
30(NE)
30(SE)
Ratio:
Injury Laboratory - 1
ox i dant
Laboratory - 2
Weekly accumulated plant injury or oxidant indices
6/17
140

10


10

65
20
50
15

0.06

0.06
6/24
240

90


180

330
190
300
190

0.38

0.74
7/1
280

225


175

80
200
200
205

0.81

0.74
7/8
230

310


350

200
260
255
130

1.33

1.52
7/15
215

270


300

270
295
305
310

1.26

1.38
7/22
195

400


340

235
290
250
250

2.06

1.77
7/29
220

290


255

260
235
225
200

1.31

1.16
8/5
295

200


100

100
165
245
255

0.69

0.34
8/12
255

250


245

120
165
190
105

0.97

0.97
8/19
165

255


330

305
270
325
225

1.58

2.04
8/26
100

190


190

235
160
85
175

1 .92

1 .89
9/2
195

125


125

190
175
175
255

0.66

0.66
9/9
125

190


130

55
200
110
195

1.53

1 .04
Total
2655

2805


2730

2445
2625
2715
2510

1 .06

1.03
o
CO
>
o
o
o
O
Values are reported as average percent cumulative new leaf injury per plant (average of four plants)..  Each,plant value'is

obtained by adding the injury percentages from each leaf.  Differences, which might be due to area distribution of oxidant

and/or to localized meteorological conditions, do occur among locations.



The oxidant index is the accumulation of hourly averages that exceed 3.0 pphm of oxidant (Kl) during the hours 6 a.m. to

10 p.m.  When the oxidant level  increases to 3-0 pphm, all  hourly averages are Included through the last one above 3.0 pphm,

even if some of the middle values fall below 3.0 pphm.  The total is then divided by 2 to obtain the oxidant index.

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                             Table 8.  RESULTS OF SECOND CINCINNATI  AREA SUMMER MONITORING PROGRAM3
Location
Oxidant
(laboratory)13
Laboratory
Benken
Matthis
Garrison
Shoemaker
Distance (miles)
and direction
from Cincinnati
5(E)
5(E)
7(NE)
25(E)
50(E)
75(E)
Ratio:
Injury , .
OxTdaTt Laboratory
Weekly accumulated plant injury or oxidant indices
6/13
310
10
40
15
10
0

0.03
6/20
200
60
125
65
65
25

0.30
6/27
90
50
200
130
240
85

0.56
7/5
35
200
185
215
230
285

5.70
7/12
110
85
110
340
255
125

0.77
7/19
40
230
120
80
115
260

5.80
7/26
80
95
75
80
65
225

1.19
8/2
210
270
255
265
205
140

1 .29
8/9
170
290
260
225
255
270

1.71
8/16
135
180
240
205
75
205

1.33
8/23
150
110
160
140
75
55

0.74
8/30
95
200
80
115
120
120

2.10
9/5
110
175
135
280
140
230

1.59
Total
1735
1955
1985
2155
1850
2025

1.13
Values are reported as average percent cumulative new leaf injury per plant (average of four plants).   Each plant value is
obtained by adding the injury percentages from each leaf.  Differences,  which might be due to area distribution of oxidant
and/or localized meteorological conditions, do occur among locations.

The oxidant index is the accumulation of hourly averages that exceed 3.0 pphm of oxidant (Kl) during the hours 6 a.m.  to
10 p.m.  When the oxidant level increases to 3.0 pphm, all hourly averages are included through the last one above 3-0 pphm,
even if some of the middle values fall below 3.0 pphm.  The total is then divided by 2 to obtain the oxidant index.

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      No consistent relationship between oxidant values and plant injury was
found during the 1967 monitoring season.   The data suggest that within the
limits of the study little difference exists  in the phytotoxic potential of the
pollution complex with distance from Cincinnati.  During any given week
there are real differences  in amount of injury among sites. It is suggested
that Cincinnati is not the major source of pollution for the  two distant sites
and may have relatively little effect on the site farthest from the city.  How-
ever, no area of southeastern Ohio is free of phytotoxic oxidant pollution.
Considerable new injury was recorded during each week of the  test period.
 14

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                             DISCUSSION

      The technique for a relatively simple plant monitoring system for the
photochemical complex has been developed.  The  system employs the sensi-
tivity of Bel W3 variety tobacco grown under controlled soil conditions and
50 percent shade.  Methodology was worked out and duplicated over two
seasons of actual exposures.  Although correlations with oxidant levels are
poor, the injury resembles ozone  injury to tobacco under controlled exposures
and the field injury is associated with the photochemical complex.

      Results presented in this paper suggest that the impact of the photo-
chemical complex is not just an urban problem but is  a problem in rural
areas as well.  Probably all areas east of the Mississippi have sufficient
photochemical pollution to produce injury to sensitive plants at certain times
during their development.

      The use of this  monitoring system would give a community estimates
of the frequency of the occurrence of phytotoxic levels of oxidants, an esti-
mate of severity of each fumigation, and some estimate of areal distribution
of phytotoxic potency in both time  and space.
                                    15

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                            APPENDIX
 TOBACCO  MONITORING  PROGRAM PROCEDURE

I.  Plants
   A.   Tobacco
       1.  Scientific name - Nicotiana tabacum,  L.
       2.  Variety  Bel W3 (from Dr.  H. E. Heggestad,  USDA,  Beltsville,
          Maryland).

   B.   Culture
       1.  Seedlings and transplants must be grown in an enclosure in which
          the air is charcoal filtered.

       2.  Seeding

          a. Fill a 4-inch pot with vermiculite and wet the vermiculite.
          b. Spread 30 to 50 seeds over the top of the vermiculite.  This
            will yield about 30 good seedlings per pot.

          c. Place pot in a  saucer.
          d. A special growth chamber in which temperature can be regula-
            ted (70°F night temperature and  80°F day temperature) and
            day length can be  regulated (8-hour day with a light intensity
            of 2000 ft-c) is recommended for growing seedlings.
          e. Water seedlings pot from the bottom (in the  saucer) with 1/2-
            strength nutrient (see III) until after  emergence.
          f. After emergence,  seedlings  can be watered from the top with
            1/2-strength nutrient.

       3.  Trans planting

          a.. When plants are 25 to 30 days  old, transplant into individual
            4-inch pots in  a peat-perlite mix (see II-A).

            (1)   Separate plants carefully,  allowing some vermiculite to
                 remain on the roots.

            (2)   Hold the plant by the  stalk in the center of an empty 4-inch
                 pot and add the peat-perlite mix.  The mix should be
                 packed gently to provide good  contact with roots.  The mix
                 should be  added to within 1/4-inch from the top of the pot
                 and  the plant held at a height so  that the leaves are exposed
                 to the  air.

            (3)   Wash mix from the leaves with distilled or deionized water.
            (4)   Soak each pot with full-strength  nutrient (100   150 cm3)
                 immediately after transplanting.

            (5)   Place  pots into trays to which full strength nutrient is
                                   17

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                   added.   The trays should be washed thoroughly at least
                   once a week to control algae growth and be refilled with
                   full-strength nutrient.  Distilled or deionized water can
                   be added to trays when needed.
              (6)   These transplants are grown in the greenhouse where the
                   temperature  is 80° to 95 °F during the day and 70 °F at
                   night.
              (7)   After the seedlings have been transplanted into individual
                   pots  and have three good-sized leaves (about 2 weeks  after
                   transplanting), remove any lower leaves less than 5 inches
                   in length (measured from the stalk to the  tip of the leaf).

           b. At this stage of growth (at least 3 good-sized leaves),  trans-
              plant the  tobacco  into bushel baskets.

              (1)   Fill the baskets to approximately 2 inches from the top
                   with the peat-perlite-soil mix (see II-B).
              (2)   Remove plant with the peat-perlite mix from the 4-inch
                   pot and  place it into a hole made in the soil.   Pack the
                   soil firmly around the plant and moisten the soil in the
                   basket with 2 gallons of  20-20-20  fertilizer solution (see
                   I-B-5-a (2).

        4. Use four plants per location.

           a. After  the first 4  plants have been in baskets for 3 weeks,
              replace two  of them.
           b. Thereafter,  every 3 weeks replace the two oldest  plants.

        5. Daily care

           a. Recommended watering schedule

              (1)   Every other week with Cal-Mag Special.

                   (a)  Mix 1 tablespoon of the fertilizer per 2 gallons of
                       •water.

                   (b)  Water each plant with 2 gallons of the solution.

              (2)   Alternate  20-20-20 with Cal-Mag every other week.

                   (a)  Mix 1 tablespoon of  the fertilizer per 2 gallons of
                       water.

                   (b)  Water each plant with 2 gallons of the solution.

              (3)   Water plants  with tap water at least twice a week in addi-
                   tion to the nutrient.  Soak soil until water comes out base
                   of the basket (2 gallons).

                   (a)  Soil must be kept moist at all times or tobacco will
                       lose sensitivity.

                   (b)  Peat mulch may be used to help keep  the  soil moist.
18                                                    TOBACCO, A SENSITIVE MONITOR

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           b. Disease and insect control

              (1)  There is a need for a general spray program for insect
                  control.

                  (a)  Chlordane sprayed in the basket area (not on the plant)
                      will offer some help.
                  (b)  Plants may be sprayed with Isotox and Malathion
                      (each mixed at the rate  of 1 tablespoon per gallon of
                      tap water) whenever insects appear, or  on a regular
                      schedule.
                  (c)  A systemic insecticide may be used in place of the
                      spray.  Add every month.

              (2)  If a plant becomes  diseased, discard plant and basket.

II.  Soil

    A.  Peat-perlite-mixture

        1. Materials needed

           a. Peat moss
           b. Perlite
           c. Lime (superfine limestone) and CaSO4 or gypsum
           d. Distilled or deionized water

           e. 10-quart pail
           f.  Small cement mixer  preferred method of mixing

        2. Procedure   mix in cement mixer

           a. 2 pails (10 qts. each) of peat moss (level full)
           b. 2 pails (10 qts. each) of perlite (level full).   To each pail of
              perlite add 2000 cm^ of distilled or deionized water until all
              perlite is  wet.  This  equals 4000 cm^ per mix.
           c. When the peat moss and perlite are well mixed (approximately
              5 minutes) add:  72 grams lime and 48 grams CaSO4 or gypsum.
              Add slowly by hand (tends to stick to sides of mixer).

           d. Mix for 20 to 30 minutes, stopping mixer two or  three times to
              break up lumps.
           e. Let mix age for 2 weeks and check pH.  Mix  pH should be
              between 5. 5 and 6. 0.  If pH is  below 5. 2 remix using another
              72 grams  of lime.  This should bring the pH  to above 5.5.

    B.  Peat-perlite-soil mixture for bushel baskets

        1. Use three parts  peat-perlite with one part top soil.
        2. Rich loam soil is recommended.

        3. Mixing is done in cement mixer following the same general
Appendix                                                                  19

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            outline as listed for mixing the peat-perlite but with no water or
            mineral additions.   The pH should be between 5. 5 and 6. 5.

III. Nutrient solution (Hoagland's) for plants  in 4-inch pots. All of the  chemi
    cals needed for making the nutrient should be mixed with distilled or
    deionized water to form concentrated solutions.  From these concentra-
    ted solutions,  use the required  amounts needed to make the nutrient.
    Technical grade KNOs, Ca(NC>3)2. 4H2O), MgSO4. 7H2O, KH2PO4,  and
    K2HPO4 is sufficient.
    Chemical
    KNO3
    Ca(N03)2

    MgSO4. 7H2O
    KH2P04

    K2HP04

    Fe chelate

    Solution B
    {Minor  elements)
Grams per liter  of
   concentrated
     solution
 101

 200

 246
 lOZjMix these two to
    /•form one concen-
 43
   /tration
                                                         cm^ of concentrate
                                                         per liter of water
                                                        (for nutrient solution)
                                                               0. 05 gm
Grams per liter of
concentrated
s olution
2
1
0
0
87
80
22
08
These form
one concen-
trated
solution
                                                         cm^ of concentrate
                                                          per liter of water
    Chemical
    H3B03
    MnCl2.4H2O
    ZnSO4. 7H2O
    CuSO4. 5H2O

          1/2-strength nutrient   equal parts of distilled or
          deionized water and full-strength nutrient solution.

IV. Enclosure     frame for housing 4 plants
    A.  8 by 4 by 6 feet high
    B.  Recommended construction

        1. Use 1- by 4-inch lumber

        2. If the program is to continue for one year,  redwood should be
           used.

        3. Brace corners with metal braces and bolts.
        4. Frame should be well anchored.

    C.  Shading

        1. North side open - all other sides covered with shade cloth
        2. 50 percent shade needed
 20
                                              TOBACCO, A SENSITIVE MONITOR

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        3. Shade material

           a. Saran shade material of 52 percent calculated shade recom-
              mended.
           b. Comparable material may be substituted for the saran.

        4. Location

           a. Open area - no natural  shade  south exposure  on hills

           b. Have access to water
           c. Protect from possible vandalism
           d. Plants must be arranged in the enclosure so all have same
              exposure.

 V. Injury indices

    A.  Reading

        1. All plants should be read weekly, daily, or on some regular
           schedule.
        2. The same individual should do all the reading.

        3. The percentage of injury on each leaf of each plant should be
           determined.   This is a subjective measure.

    B.  Summary

        1. The percentage of area injured  on each leaf is totaled for each
           plant at every reading.
        2. New injury on each plant for  any given period is determined by
           the difference in  cumulative readings between the beginning and
           end of the periods.
        3. Site averages are obtained by summing the four plant values and
           dividing by four.
Appendix                                                                 21

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                             REFERENCES

1.  Heck, W. W. , The use of plants as indicators of air pollution, Int. J. Air
   and Water Poll.,  1_0:99-111 (1966).

2.  Noble,  W. M.and L. A. Wright, A bio-assay approach to the study of air
   pollution, Agron. J. ,  50^:551-553 (1958).

3.  Middleton, J. T.  and A.  O. Paulus, The identification and distribution of
   air pollutants through plant response,  Arch. Ind.  Health,  14:526-532
   (1956).

4.  MacDowall,  F.  D. H. , E.  I. Mukammal, and A. F.  W. Cole,  Direct
   correlation of air-polluting ozone and  tobacco weather fleck, Can. J.
   Plant Sci. ,  44:410-417 (1964).

5.  Heggestad,  H. E. and H. A.  Menser,  Leaf spot-sensitive tobacco strain
   Bel W3, a biological indicator of the air pollutant ozone, Phytopathology,
   52:735  (1962).
                                    23
                                     GOVERNMENT PRINTING OFFICE : 1969 O - 353=080

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