Biomass 9 (1986) 239-246

Effect of Stage of Development on Chemical Yields in

Common Milkweed, Asclepias syriaca
T. A. Campbell and Karla A. Grasse
Germplasm Resources Laboratory, Plant Genetics and Germplasm Institute,
Agricultural Research Service, US Department of Agriculture, Beltsville, MD 20705,
USA
(Received: 26 October, 1985)

ABSTRACT
Common milkweed has been identified as a potential whole-plant
source of polyphenols, oil, and natural rubber for industry. The effects
of stage of development (prebud, full bud, full flower, seed set, and
mature seed) on yields of acetone extract ('polyphenols' and 'oil') and
hexane extract (natural rubber) were studied in two Maryland ( USA )
populations in 1982 and 1983. Leaf fraction and stem fraction acetone
extract yields were generally highest at the mature-seed stage whereas
stem fraction hexane extract yields were highest at the prebud stage in
1982 and at the full-bud stage in 1983. There was generally a strong
direct relationship between yields of leaf fraction acetone extract and
leaf fraction hexane extract and a strong inverse relationship between
yields of stem fraction acetone extract and stem fraction hexane extract.
Results indicate that environmental variation can have a substantial
effect on levels of polyphenols, oil and natural rubber at a particular
stage of development; however, it appears that harvesting at the manureseed stage of development would usually maximize yields of both
extracts.
Key words: Polyphenols, oil, natural rubber, nonrenewable raw
materials.

INTRODUCTION
B u c h a n a n and O t e y t p r o p o s e d the d e v e l o p m e n t of multi-use perennial
crops that would be whole-plant sources of polyphenols, oil, and natural
r u b b e r (NR) thus reducing the d e m a n d for n o n r e n e w a b l e industrial raw
239
Biomass 0144-4565/86/S03.50 - Elsevier Applied Science Publishers Ltd, England,
1986. Printed in Great Britain

240

T. A. Campbell, K. A. Grasse

materials. They identified common milkweed (Asclepias syriaca L.) as a

promising species.
Natural stands of common milkweed sampled near Peoria, Illinois
(USA) in September and October of 1975 contained approximately
7.2% 'polyphenols', 4.3% 'oil', and 1.4% NR on a dry weight basis. 2 In 48
populations sampled between 12 June and 27 June, 1979, 3 extract yields
on a dry weight basis ranged from 4.7 to 14.4% for acetone extract ('polyphenols' and 'oil'), and 0.2-1.2% for hexane extract (natural rubber).
Gerhardt 4 determined that most of the natural rubber in common milkweed occurred in leaves and ranged from 0.5% of the dry weight in May
to 2.9% in October. The polyphenol fraction is composed of polyphenolics, phylobaphenes, tannins, and complex lipids. 2 In an evaluation
of common milkweed from wild stands, 1 5% of the 'oil' fraction was
sterols, 11% other free alcohols, 5% free acids, 72% nonglyceride esters,
7% hydrocarbons, and only a trace was triglycerides.
Polyphenols in common milkweed have not been fully characterized,
but polyphenols in general have potential for use in wood laminating
resins, plywood glues, particleboard adhesives, fortifiers for starch
adhesives, oil well drilling muds, clay flocculants, plastics formulation,
antioxidants and controlled release formulations.~ Oils in common milkweed have the potential to produce a wide variety of chemical intermediates; a substantial market may exist for oils that are rich in
nonglyceride esters for use as extender oils and/or processing aids for
rubber and as plasticizers for plastics.J Whole-plant oils, and to a lesser
extent polyphenols, may also be able to compete in the petroleum
market as fuels and basic raw materials if petroleum prices continue to
rise.
Proton magnetic resonance spectra for the hydrocarbon fraction of
common milkweed and Hevea brasiliensis natural rubber are essentially
identical;2 however, the molecular weight of common milkweed NR is
substantially lower than that of Hevea NR. ~ Lower molecular weight
NRs could be used as plasticizers and processing aids for Hevea and
synthetic rubbers, as liquid or easy-processing rubbers for special uses,
and in adhesive formulations; there could be an unlimited market for
lower molecular weight NR which competed in price with petroleum.
Extract-free residue from common milkweed has potential for use as a
primary or supplemental cattle feed, as a fermentation substrate, or in
the production of methane, charcoal, or wood distillation products; bast
fiber is of good quality and appears to have potential for papermaking.2
Common milkweed can be direct seeded after breaking dormancy
with GA 3 and sodium hypochlorite3 and could probably be harvested
twice yearly beginning the year after seeding (Campbell, unpublished

Effect of stage of development on yields in Asclepias syriaca

241

data). Buchanan and Otey ~ suggested that it would be economically

feasible to grow milkweed as a crop if dry-matter yields were at least
13"5 Mg ha -1 yr -1 and dry matter contained 10% NR, 6% oil and 7%
polyphenols. These yields probably cannot be achieved without substantial modification of the species through breeding and through the
development of effective management techniques.
Data on the effects of stage of development on chemical yields are
needed before effective management techniques can be developed. The
following experiment was conducted to study the in situ changes in
acetone and hexane extract yields in two common milkweed populations
during growth and development.

MATERIALS AND METHODS

Two Beltsville, Maryland (USA) populations approximately 3.7 km
apart and located on Interstate Highway 95 were chosen. Population 1
(P1) was located on a steep embankment and covered an area of approximately 135 m2; plants were 0.6-0.9 m in height. Population 2 (P2) was
located on a gentle slope and covered an area of approximately 1281 m2;
plants were 0"9-1.2 m in height. Both were located on disturbed land
with good drainage.
Each population was divided into four blocks of equal area. Four
plants per block were harvested at the prebud, full bud, full flower, seed
set, and mature seed stages of development during the summers of 1982
and 1983. Plants sampled were at least 1 m apart. Sampling dates are
given in Table 1.
Material from each block was dried to approximately 5% moisture at
ambient temperatures (25-35C), partitioned into leaves and stems, and
stored at 10C with 40% relative humidity. Each fraction was ground in a
rotary mill (0"4 mm screen) and extracted (in triplicate, 5 g per extraction) with acetone then hexane for 4 h each using a Goldfisch apparatus.
Mean % acetone and % hexane extract (designated AEX and HEX,
respectively) on a dry weight basis were computed for each fraction.
An analysis of variance was conducted on AEX and HEX for leaf and
stem fractions after transforming data to square roots to improve
symmetry.5 Years (Y) and blocks (B) were assumed to be random and
stages of development (S) fixed. Sources of variation were partitioned as
shown in Table 2. Because blocks for P1 had to be relocated in 1983, the
B(Y) mean square was used to compute F-values for testing P1 Y effects
whereas P2 Y effects were tested with the residual mean square. For all

242

T. A . Campbell, K. A. Grasse
TABLE 1

Mean % Acetone and % Hexane Extract Yields" from Two Maryland (USA) Common
Milkweed Populations Based on in situ Sampling in 1982 and 1983
Year

Stage of
development

Sampling
date

Leaf fraction

Stem fraction

Acetone
extract

Hexane
extract

Acetone
extract

Hexane
extract

1982
1982
1982
1982
1982
1983
1983
1983
1983
1983

Prebud
Full bud
Full flower
Seed set
Mature seed
Prebud
Full bud
Full flower
Seed set
Mature seed

15 June
18 June
12 July
22 July
25 August
26 May
9 June
28 June
25 July
19 August

Population 1
10.19
9'00
9.40
11-23
11.14
8.64
8'01
9-41
10-35
12.36

1.09
0.75
0.96
1-44
1.30
0-44
0-63
1-04
2-59
2.80

5.73
4.80
6-45
5.41
7.35
5.67
6"17
5.25
6.42
7.55

0.18
0"10
0-16
0.16
0.08
0.28
0-46
0.29
0-27
0.27

1982
1982
1982
1982
1982
1983
1983
1983
1983
1983

Prebud
Full bud
Full flower
Seed set
Mature seed
Prebud
Full bud
Full flower
Seed set
Mature seed

15 June
18 June
12 July
22 July
25 August
26 May
9 June
28 June
25 July
19 August

Population 2
10.14
8-86
11.52
10.92
11.56
8.64
8"55
7"36
10-22
10-87

0.78
0.86
1-68
2.50
3.34
0.52
0"58
1'20
1.86
3-70

5"89
5-11
7.16
5-46
6.19
5.79
4"99
4.95
5.81
5-83

0.21
0.14
0-10
0.17
0.20
0.34
0.38
0-36
0.31
0-17

"Dry weight basis.

analyses, S effects w e r e t e s t e d with the Y x S m e a n s q u a r e a n d Y x S

effects w e r e t e s t e d with the residual m e a n square.
F o r e a c h y e a r a n d p o p u l a t i o n , all t r a n s f o r m e d r e s p o n s e variables w e r e
c o r r e l a t e d a n d t h e s i m p l e c o r r e l a t i o n m a t r i x was r e d u c e d using p r i n c i p l e
c o m p o n e n t (PC) analysis. 6'7 P C ' s that did n o t r e p r e s e n t at least the variation d u e to o n e v a r i a b l e ( e i g e n v a l u e o f at least 1 ) w e r e d e l e t e d as r e c o m m e n d e d b y H a r m a n . 6 P C analysis r e d u c e s a c o r r e l a t i o n m a t r i x to a series
o f c o l u m n v e c t o r s (called PC's) r e p r e s e n t i n g the r e l a t i o n s h i p s a m o n g the
variables. T h e c o e f f i c i e n t s o f the P C ' s r a n g e f r o m - 1 to 1 a n d are called
'loadings'. Variables t h a t l o a d highly o n a P C are highly c o r r e l a t e d ; high

Effect of stage of development on yields in Asclepias syriaca

243

TABLE 2

Mean Squaresa from an Analysis of Variance of the Effects of Stage of Developmenton

Acetone Extract and Hexane Extract Yields from Two Maryland (USA) Common Milkweed Populations Based on in situ Samplingin 1982 and 1983
Source of variation

df

Lear fraction
Acetone
extract

Hexane
extract

Stem fraction
Acetone
extract

Hexane
extract

Years (Y)
Stage of development(S)
Yx S

1
4
4

Population 1
0"11
0.09
0.33
0.65
0"05
0"28**

0"05
0.17
0"05

0"27**
0.02
0"02

Years (Y)
Stage of development(S)
Yx S

1
4
4

Population 2
0'55**
0"22
0-13"*

0-07
0"06
0"08

0"33**
0"02
0.02

0.26
1-64"*
0"03

Response variables were transformed usingthe formula (% extract)~2.

**Indicates significanceat the 1% level.

loadings with like signs indicate that the variables they represent are
positively correlated whereas high loadings with opposite signs indicate
negative correlation. PC's are often post-multiplied by a transformation
matrix which rotates them in PC space in an effort to force each variable
to load highly on only one PC. For these data, a Varimax rotation 6,7 was
used. This procedure attempts to simplify the colunms of a PC matrix by
maximizing the variance of the squared loadings in each column.

RESULTS A N D DISCUSSION
For P1, year effects were significant only for stem fraction HEX, stage of
development effects were nonsignificant for all response variables, and
the years x stage of development interaction was significant only for leaf
fraction H E X (Table 2). For P2, year effects were significant for leaf fraction A E X and stem fraction H E X whereas stage of development and the
years x stage of development interaction were only significant for leaf
fraction H E X and leaf fraction A E X , respectively.
Leaf fraction A E X and H E X were highest at the mature-seed stage
except for P1 in 1982 where they were both highest at the seed-set stage
and second highest at the mature-seed stage (Table 1 ). In P2, leaf fraction

T. A. Campbell, K. A. Grasse

244

AEX was consistently greater in 1982 than in 1983 at all stages of

development and was substantially greater at the prebud and full-flower
stage in 1982; leaf fraction HEX yields increased steadily during the
growing season both years in P2. Based on Scheffe's test at the 5% level,5
leaf fraction HEX was significantly greater at the seed-set and matureseed stages than at the prebud and full-bud stage in P2. Although variation in stem fraction AEX was nonsignificant, yields were highest at the
mature-seed stage except for P2 in 1982 where they were highest at full
flower and second highest at the mature-seed stage. Stem fraction HEX
yields were erratic during the growing season but were generally higher
in 1983 than in 1982.
Precipitation in Beltsville for May, June, July, and August was 5-5,
12.6, 6"6, and 4.4 cm, respectively in 1982 and 13.3, 20.9, 1.6, and
8"7 cm, respectively in 1983. Mean temperatures for May, June, July,
and August were 18.1, 20.8, 24.4, and 22.0C, respectively in 1982 and
16.3, 21.9, 24.9, and 24-9C, respectively in 1983. Precipitation was
abnormally high in May and June of 1983 and was probably low enough
in July of 1983 to cause drought stress; it is doubtful if temperature stress
was a factor either year. Abnormally high precipitation in the spring of
1983 appears to have accelerated development and may have stimulated
the accumulation of stem NR in both populations and, in P2, reduced the
accumulation of leaf polyphenols and oil. The effect of abnormally high

TABLE 3
Loadings from a Principal Component Analysis of Data from a 2-year in situ Evaluation
of the Effects of Stage of Development on Acetone and Hexane Extract Yields from
Two Maryland (USA) Common Milkweed Populations

1983 Population

1982 Population

Leaf fraction acetone

extract
Leaf fraction hexane
extract
Stem fraction acetone
extract
Stem fraction hexane
extract

Factor

Factor

Factor

Factor

0"80

-0-09

0.82

0.46

0.86

0.09

0-28
0.29

0.95

0.11

0"58

0.59

0.96 - 0 " 0 5

0.93

-0.20

0"78

0-09

0.83

0.10

-0.75

-0.75

- 0.11

-0.77

0-04

0"89 - 0 . 1 8
-0.08

0.94

0.77 - 0 . 7 0

-0.17

Effect of stage of development on yields in Asclepias syriaca

245

precipitation on leaf polyphenol and oil production may have been more
pronounced in P2 because more moisture percolated into the soil in
this population. It appears that drought stress in July of 1983 had little
effect on polyphenol, oil, or natural rubber production.
Two PC's were retained for each of the four analyses (Table 3). There
was a strong direct relationship between leaf fraction A E X and leaf fraction H E X and a strong inverse relationship between stem fraction A E X
and stem fraction H E X , except for P2 in 1983, where instead a strong
inverse relationship between leaf fraction and stem fraction H E X levels
was indicated. It seems that in the leaf, the ratio of assimilate channeled
to polyphenol and oil production to that channeled to NR production
may remain fairly constant, whereas in the stem, assimilate may be
channeled to the production of one of the fractions at the expense of
another at different stages of development. Perhaps the heavy spring
precipitation in 1983 altered assimilate partitioning patterns in P2
causing leaf and stem NR production to be favored alternately.

CONCLUSIONS
It is obvious that environmental variation over the years can have a substantial effect on yields of A E X and H E X at a particular stage of
development. However, considering the fact that leaves yielded the most
extract, it appears that harvesting at the mature-seed stage of development would generally maximize extract yields. It would be worthwhile to
compare extract yields from harvesting at the mature-seed stage to those
from two harvests per season at the full-flower stage; the latter system
would undoubtedly increase dry matter yields and obviate the problem
of volunteer seed dispersal. Flowers and seed pods were not analyzed in
these experiments, but their contribution to extract and dry matter yields
should certainly be examined in future experiments.

REFERENCES
1. Buchanan, R. A. & Otey, F. H. (1979). Multi-use oil- and hydrocarbonproducing crops in adaptive systems for food, material, and energy production. Biosources Dig., 1,176-202.
2. Buchanan, R. A., Cull, I. M., Otey, E H. & Russel, C. R. (1978). Hydrocarbon and rubber-producting crops: Evaluation of US plant species. Econ.
Bot., 32, 131-45.
3. Campbell, T. A. (1983). Chemical and agronomic evaluation of common
milkweed, Asclepias syriaca. Econ. Bot., 37, 174-80.

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T. A. Campbell, K. A. Grasse

4. Gerhardt, F. (1929). Propagation and food translocation in the common

milkweed. J. Agric. Res., 39,837-51.
5. Snedecor, G. W. & Cochran, W. C. (1967). Statistical methods, 6th edn, Iowa
State Univ. Press, Ames, Iowa.
6. Harman, H. H. (1976). Modern factor analysis, 3rd edn, Univ. Chicago Press,
Chicago, Illinois.
7. Ray, A. A. (1982). Statistical analysis system users guide: Statistics, 1982 edn,
SAS Institute, Raleigh, North Carolina.