Natural and environmentally
friendly way of managing
weeds without herbicides
by Allelopathy
Yoshiharu FUJII,
Tokyo University of Agriculture and Technology,
2.50pm-3.20pm, Monday 24 February 2014
Crown Perth, Burswood
1
Outline of Talk
What is Allelopathy
• Selection of allelopathic plants by Bioassays
• New Concept of Total Activity for the isolation
of Allelochemicals.
• Cover crop with allelopathic activity provides
natural and environmentally friendly way of
managing weeds without herbicides.
2
Allelopathy
Interaction (inhibitory or stimulatory)
between plants or plant to other life
by natural chemicals
(allelochemicals)
Plant to Plants ・・・・・・・・・・weed control
Significance of Secondary metabolites*
Plant
Microorganisms・・・disease
(*notto
indispensable
natural chemicals in plants) control
Plant to Insects ・・・・・・・・・・ harmful Insect control
Plant to Animals ・・・・・・・・・・nematode, boar, etc.
Allelopathy is the same concepts as “Antibiotics”
3
Bioassay for Three Route
All these Bioassays are
my original Assay
1) Exudation (from root)
 Plant Box Method
3)
2) Leaching (from leaf
or litter)
2)
 Sandwich Method
2)
3) Volatilization (mainly
from leaf as volatile
chemicals)
 Dish Pack Method
1)
4)=1)’ Rhizosphere Soil Method
Fig. 1 Route of Allelopathy and Bioassay 4
4
1) Specific Bioassays
for Allelopathy
For the screening of
Allelopathic Plants
5
2.1 Plant Box Method
for root exudates (Mixed planting)
Root
zone
separating
tube
Radicle Length of Acceptor (mm)
80
60
40
20
y = 1.3355x + 14.85
2
R = 0.9066
0



0
10
20
30
40
Sand Culture for１-２ month
Distance from the Root of Donor (mm)
Plant Box (for tissue culture)
Dr. Fujii,
Agar Medium (no nutrients)
Fujii, Y. et al., Plant-Box Method: A Specific Bioassay to Evaluate
Allelopathy through Root Exudates. Allelopathy, New Concepts and
Methodology, Science Publisher, 39-56 (2007).
50
1991
1
6
Table 1.
Allelopathic activity Leguminosae Family by Plant Box Method
Scientific name
Pachyrhizus erosus
Mucuna pruriens var. utilis
Mimosa invisa
Crotalaria zanzibarica
Medicago scutellata
Galega orientalis
Coronilla varia
Medicago arabica
Trifolium album
Medicago lupulina
Phaseolus vulgaris
Vicia villosa var. dasycarpa
Abrus praecatorius
Canavalia ensiformis
Lupinus bicolor
Vicia villosa var. villosa
Melilotus albus
Dolicos lablab
Vicia sativa
Pueraria lobata
Crotalaria juncea
Trigonella foenum-graecum
Vigna unguiculata
Cassia tora
Medicago sativa
Vicia faba
Onobrychis viciifolia
Pachyrhizus tuberosus
Phaseolus vulgaris
Vigna angularis
Trifolium incarnatum
Cajanus cajan
Crotaralia spectabilis
Pisum sativum
Cicer arietinum
Lathyrus odoratus
Amorpha fruticosa
Latyrus sativus
Lupinus albus
Arachis hypogaea
Calopogonium mucunoides
Desmodium paniculatum
Vigna unguiculata subsp. sesquipedalis
Mimosa pudica
Vicia angustifolia var. segetalis
Aeschynomeue rudis
English name
Jicama
Velvetbean
activity
6
7
9
12
13
14
16
16
16
19
19
19
20
20
20
20
23
25
25
28
30
30
33
34
34
34
35
36
37
38
39
40
40
42
44
44
45
46
46
49
49
49
49
55
55
58
Allelopathic
Leguminous
Plants
Sunn hemp
Snail medic
Galega
Crown vetch
Hop clover
Kidney bean (Old cv.)
Woolly pod vetch
Rosary pea
Jack bean
Hairy vetch
White sweet clover
Lablab bean
Common vetch
Kudzu
Sunn hemp
Fenugreek
Cowpea
Sickle senna
Alfalfa
Broad bean
Esparcette
Ahipa
Kidney bean (Modrn cv.)
Adzuki bean
Crimson clover
Pigeon pea
Sunn hemp
Pea
Chickpea
Sweet pea
False indigo
Grass pea
White lupine
Peanut
Calopogonio
Tick trefoil
Asparagus pea
Sensitive plant
Karasunoendou
Joint vetch
Pachyrhizus
Mucuna
Mimosa
Crotalaria
Medicago
Vicia
Plant
Box
Method
Mucuna
Mimosa
7
Table Allelopathic activity of Poaceae family evaluated by Plant Box Method (selected results)
Scientific name
Hordeum vulgare (Barley, cv. HVG-1)
Avena strigosa (Wild Oat)
Triticum polonicum (Polish Wheat)
Avena sativa (Oat, Hokuren)
Avena murphyi (Wild Oat)
Secale cereale (Rye, Samusa-shirazu)
Avena barbata (Wild Oat)
Triticum compactum (Club Wheat)
Avena longiglumis (Wild Oat)
Triticum spelta (Spelta Wheat)
Triticum sp. x Secale sp. (Triticale)
Avena orientalis (Wild Oat)
Chloris gayana (Rhodesgrass)
Avena wiestii (Wild Oat)
Avena byzantina (Byzantine Oat)
Hordeum vulgare (Barley, Sansyu)
Phalaris tuberosa (Hardinggrass)
Triticum aestivum (Common Wheat, Mulchmugi)
Anthoxanthum odoratum (Sweet Vernalgrass)
Festuca rubra (Chewing Fescue)
Panicum coloratum (Coloured Guinea Grass)
Avena fatua (Common wild oat)
Triticum aestivum (Common Wheat,cv. Norin 61)
Radicle
growth
9.7
12.0
13.2
14.8
16.9
17.3
18.3
19.5
21.8
22.4
22.6
22.8
23.5
24.1
24.5
25.5
26.6
28.0
28.1
30.3
32.8
33.0
33.5
Scientific name
Radicle
growth
Scientific name
Allelopathic Crops
Eleusine coracana (Finger Millet)
Triticum monococcum (Einkorn Wheat)
Sorghum bicolor (Sorghum, Shikoku-native)
Coix lacryma (Coix)
Panicum kabulabula (Kabulabula Grass)
Avena sativa (Oat, cv. Hayoats)
Triticum durum (Durum Wheat)
Avena abyssinica (Ethiopian Oat)
Avena brevis (Brevis Oat)
Panicum maximum (Guinea Grass)
Bromus catharticus (Rescuegrass)
Triticum dicoccum (Emmer Wheat)
Sorghum halepense (Johnson Grass)
Panicum antidotale (Blue Panicgrass)
Eragrostis tenuifolia (Eragrostis)
Festuca arundinacea (Kentucky 31 fescu)
Sorghum sudanense (Sudan grass)
Agropyron repens (Quackgrass)
Lolium subulatum (Wimmera Ryegrass)
Sorghum dochna (Common sorghum)
Zea mays (Corn, cv. Peter corn)
Setaria viridis (Foxtail Grass)
Pennisetum glaucum (Pearl Millet)
33.7
34.3
35.5
36.5
36.7
37.0
37.3
38.5
38.8
39.7
40.1
40.5
42.8
43.5
44.6
44.8
45.2
46.0
46.8
47.1
47.7
53.2
54.4
Echinochloa utilis (Green Millet)
Euchlaena mexicana (Teosinte)
Chloris distichophylla (Weeping Chloris)
Zea mays (Corn, cv. Golden dent)
Panicum virgatum (Switch Grass)
Poa pratensis (Kentucky Bluegrass)
Panicum dichotomiflorum (Fall panicgrass)
Eragrostis lehmanniana (Lehmann Lovegrass)
Dactylis glomerata (Orchard Grass)
Eragrostis curvula var. valida (African Lovegrass)
Agrostis alba (Red Top)
Lolium perenne (Perennial Ryegrass)
Zea mays (Corn, cv. Pioneer dent)
Cenchrus ciliaris (Buffel Grass)
Paspalum dilatatum (Dallis Grass)
Paspalum notatum (Bahia grass)
Eleusine indica (Goose Grass)
Lolium multiflorum (Italian Ryegrass)
Poa compressa (Canada Bluegrass)
Phalaris arundinacea (Reed Canary grass)
Lolium westerwoldicum (Westerwolds Ryegrass)
Festuca elatior (Meadow Fescue)
Phleum pratense (Timothy)
Barley (Hordeum vulgare)
Oats (Avena sp.)
Rye (Secale cereale)
Wheat (Triticum sp.)
These upland crops are
originally resistant for weed
Radicle
growth
55.8
55.8
56.9
57.9
58.5
59.8
61.2
61.8
61.9
61.9
62.1
63.6
66.3
70.7
72.4
72.9
73.5
73.9
74.0
76.3
84.8
86.8
96.4
8
Jean-François Millet, Le semeur (1850)
bird
seed
We Japanese
love Millet !
Wheat wheat has allelopathic activity
amount of seeding rate = 3 to 5 times
For bird loss and weed suppression
9
“MACE” is King variety in AU now
(by Cristine Zaicou-Kunesch)
At today’s morning session
Table Allelopathic activity of Triticum family
evaluated by Plant Box Method
Scientific name
Radicle
growth (%)
Triticum polonicum (Polish Wheat)
13.2
Triticum compactum (Club Wheat)
19.5
Triticum spelta (Spelta Wheat)
22.4
Triticum sp. x Secale sp. (Triticale)
22.6
Triticum aestivum (Common Wheat, cv. Mulchmugi)
28.0
Triticum aestivum (Common Wheat, cv. Norin 61)
33.5
Triticum monococcum (Einkorn Wheat)
34.3
Triticum durum (Durum Wheat)
37.3
Triticum dicoccum (Emmer Wheat)
40.5
Proposal
for Project
Breeding of
Weed Resistant Wheat
with
high allelopathic
activity !
10
Allelochemicals reported from Oats, Rye, Wheat, Barley
O
CH3O
HO
Wheat
Rye
Oat
O
O
7-Hydroxy-5-methoxycoumarin
(Scopoletin)
Many other chemicals
are reported, but I
guess these traditional
compounds are
important as allelochemicals based on TA
N
OH
H3CO
O
N
2,4-dihydroxy1,4-benzoxazine-3-one
(DIBOA)
HO
2,4-dihydroxy-7-methoxy1,4-benzoxazine-3-one
(DIMBOA)
H3CO
O
O
N
H
2-benzoxazolinone
(BOA)
CH3
O
OH
OH
CH3
OH
N
O
Barley
N,N-dimethyltyramine
(Hordenine)
O
O
N
H
6-methoxy2-benzoxazolinone
(MBOA)
As for “TA”,
Total Activity,
Will explain later.
1. Belz, G.R. and Hurle, K., Dose-Response - A Challenge for Allelopathy? Nonlinearity Biol. Toxicol. Med. 3
(2), 173–211 (2005)
2. Liu, D. L. and Lovett, J. V., Biologically active secondary metabolites of barley. II. Phytotoxicity of barley
allelochemicals, J. Chem. Ecol. 19, 2231-2244 (1993)
11
2. Sandwich Method
for leaf leachates (Mulching, Litter)
6 well multi-dish
make sandwich by agar
10 or 50 mg d.w./10 cm2
For allelopthy by fallen leaves and litters
fallen leaves are constant
（3 ton / ha / year） = 30 mg d.w./10 cm2 Dr. Fujii, 1991
Fujii,
et al.,
Assessmentmethod
method for allelopathic
from
leafleaf
litter
leachates.
Fujii,
Y. Y.
et al.,
Assessment
allelopathiceffect
effect
from
litter
leachates.
Weed
Biology
and
Management, 4(1),
4(1), 19
– 23
(2004).
Weed
Biology
and
Management,
1923
(2004)
1
12
Table 3.
Allelopathic activity by Sandwich Method
Scientific name
Cymbopogon citratus
Derris scandens
Piper betle
Tamarindus indica
Gliricidia sepium
Sesbania grandiflora
Acacia farnesiana
Duranta repens
Diospyros mollis
Afgekia sericea
Ipomea pes-caprae
Jatropha integerrima
Melia azedarach
Citharexylum spinosum
Molineria latifolia
Passiflora coccinea
Calophyllum inophyllum
Amhersita nobilis
Cynometra cauliflora
Litchi chinensis
Radicle
0**
0**
3**
3**
5**
11*
15*
17*
23*
38
39
44
52
70
75
86
94
96
101
113
Hypocotyl
0**
0**
18**
26*
19*
45*
37
51
22
120
90
122
115
60
167
117
146
127
113
115
Radicle and hypocotyl means % growth of lettuce seedling
to the control (in agar medium). ** after the data means
inhibitory activity stronger than standard deviation value
of 60, and * means 55.
Sandwich
Method
Cymbopogon
Tamarindus
Fujii, Y. et al., Screening of 239 medicinal plant species for allelopathic activity using
the sandwich method. Weed Biology and Management, 3(4), 233-241 (2003)
13
4. Dish Pack Method
3
for volatile allelochemicals
6 well multi-dish
sealed by tape
 measure after 3 days
plant leaf (2g)
CH3
Volatile
chemicals
were
analyzed
by GC-MS
OH
β-pinene
H3C
CH3
thymol
CH3
β-myrcene
OH
cis-3-hexene-1-ol
H3C
CH2
limonene
Fujii, Y. et al., Dish pack method: a new bioassay for volatile allelopathy. In
Proceedings of the 4th World Congress on Allelopathy, August 21 -26, 2005, Wagga
1
Wagga, Australia, 493-497 (2005).
14
Allelopathic plant for biofumigation
Cleome
Cleome spinosa）
Cleome from Takii seed Co.
Ｈ
Ｎ Ｃ Ｓ
Ｃ
Ｈ
Ｈ
Methyl Isothiocyanate
Strongest activity by Dish Pack Method
Allelochemical : Methyl Isothiocyanate (MITC)
EC100 for lettuce is 18 ppm
Kill most specise of nematode
Concentration of MITC in Cleome
表２　クレオメ各部位における
ppm
メチルイソチオシアネート濃度
M ethyl
isothiocyanate濃度(ppm ) 39
Leaf
(fresh)
茎葉
39
Root (fresh)
15
さや
16
Seed pod
16
根
5
Seed
14
種子(＋水)
14
種子(
磨砕＋水)
133
Seed (macerated)
133
15
Table
Database for Allelopathic activity evaluated by PB, SW, and DP method
表８今後導入される可能性のある外来植物600種のアレロパシー活性の検定結果（抜粋）
表2.7 今後導入される可能性のある外来植物600種のアレロパシー活性の検定結果（抜粋、データは追加・変更の可能性あり）
学　名
和　名
ｲﾁﾋﾞ
ﾔｷﾞﾑｷﾞ
科　名
ｱｵｲ科
ｲﾈ科
ｲﾈ科
ﾋﾒﾇｶﾎﾞ
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ｲﾈ科
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ﾋﾛｰﾄﾞｱｵｲ
ｱｵｲ科
ﾋﾕ科
ｲﾇﾋﾞﾕ
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ﾋﾕ科
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ｲﾈ科
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ﾔｸﾞﾙﾏｷﾞｸ類
ｾﾘ科
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ｷﾝﾎﾟｳｹﾞ科
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ｲﾈ科
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ｶﾔﾂﾘｸﾞｻ科
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ｲﾈ科
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ﾅｽ科
ｺﾒｽｽｷ
ｲﾈ科
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ﾏﾂﾑｼｿｳ科
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ｲﾈ科
ｲﾈ科
SW法
ＤＰ法
PB法
総合評価
学　名
和　名
31
105
21
1
Echinochloa frumentacea
111
27
1
Echinochloa hispidula
60
119
60
0
Echinochloa oryzzoides
ﾉｹﾞﾀｲﾇﾋﾞｴ
44
97
29
0
Echinochloa utilis
36
90
24
1
Echium italicum
42
106
43
0
Echium plantagineum
ｼｬｾﾞﾝﾑﾗｻｷ
46
88
43
0
Echium vulgare
ｼﾍﾞﾅｶﾞﾑﾗｻｷ
43
100
43
0
Eclipta alba
56
106
37
0
Eclipta erecta
41
134
95
0
Eclipta prostrata
64
91
61
0
Eleusine indica
ｼｺｸﾋﾞｴ
31
88
38
0
Emex australis
63
41
109
41
0
Emex spinosa
ｲﾇｽｲﾊﾞ
47
106
50
0
Erodium cicutarium
40
107
30
0
Erysimum orientale
25
76
53
1
Eupatorium cannabinum
ﾀｲﾜﾝﾋﾖﾄﾞﾘ
31
100
60
0
Euphorbia corollata
42
86
64
0
Euphorbia cyparissias
27
100
31
0
Euphrasia rostkoviana
44
92
15
1
Festuca arundinacea
86
7
2
Festuca ovina
21
96
24
1
Fimbristylis littoralis
50
44
89
64
0
Fimbristylis milliacea
41
59
57
1
Fumaria capreolata
ﾆｾｶﾗｸｻｹﾏﾝ
96
34
1
Galium mollugo
24
93
48
0
Geranium molle
56
38
98
24
1
Geranium pratense
34
123
39
0
Geranium pusillum
23
95
35
1
Geranium pyrenaicum
32
101
16
1
Geranium robertianum
95
10
1
Geum rivale
38
88
28
2
24
Geum urbanum
79
29
0
Guizotia abyssinica
ｷﾊﾞﾅﾀｶｻﾌﾞﾛｳ
41
115
43
0
Hirschfeldia incana
ｱﾚﾁｶﾞﾗｼ
58
85
27
1
Holcus lanatus
ｼﾗｹｶﾞﾔ
58
98
18
1
Hyoscyamus niger
ﾋﾖｽ
78
105
22
2
20
Hypochaeris glabra
83
63
0
Hypochaeris radicata
ﾌﾞﾀﾅ
36
65
20
3
Ipomoea aquatica
23
28
95
22
1
Ipomoea hederacea
26
48
37
2
Ipomoea lacunosa
39
106
57
0
Isatis tinctoria
ﾊﾏﾀｲｾｲ
33
83
29
0
Juncus articulatus
40
91
84
0
Juncus bufonius
53
0
60
1
Kichxia commutata
28
11
70
1
Lathyrus latifolius
40
105
46
0
Lathyrus nissolia
20
98
47
1
Lathyrus phaseoloides
55
105
55
0
Legousia speculum veneris
ｵｵﾐｿﾞｶｸｼ
17
106
40
1
Leontodon autumnalis
100
47
0
Leontodon hispidus
31
42
95
85
0
Leontodon taraxacoides
ｶﾜﾘﾐﾀﾝﾎﾟﾎﾟ
74
91
47
0
Lepidum sativum
ｺｼｮｳｿｳ
46
69
48
1
Lespedeza striata
37
20
28
2
Leucanthemum vulgare
ﾌﾗﾝｽｷﾞｸ
15
115
28
2
Lolium multiflorum
33
85
25
1
Lolium rigidum
42
61
13
2
Lolium temulentum
39
100
59
0
Lotus corniculatus
39
90
47
0
Lupinus perennis
126
39
0
47
Lychinis viscaria
35
74
75
1
Malva alcea
62
95
77
0
Malva hirsuta
86
32
0
Malva neglecta
ｾﾞﾆﾊﾞｱｵｲ
39
107
20
2
Malva parviflora
ｳｻｷﾞｱｵｲ
16
33
63
10
2
Marubium vulgare
ﾆｶﾞﾊｯｶ
105
82
0
Matricaria discoidea
45
77
94
24
1
Matricaria inodora
44
104
38
0
Matricaria maritima
27
111
58
0
Matricaria perforata
115
82
0
Matricaria recutita
50
22
90
27
2
Matricaria suaveolens
112
83
0
Medicago lupulina
66
54
130
58
0
Medicago sativa
66
116
77
0
Melandrium album
ﾏﾂﾖｲｾﾝﾉｳ
99
71
1
26
Melandrium rubrum
90
42
1
Melilotus albus
ｼﾛﾊﾞﾅｼﾅｶﾞﾜﾊｷﾞ
23
注１）SW法は葉から溶脱する物質による活性を、DP法は葉から揮発する物質による活性を、PB法は根から滲出する物質による活性を示す。
注２）表注の数字は、検定植物レタスの生育率（％）を表しており、数値が小さいほどアレロパシーによる阻害活性が強いことを示す。
注３）判断基準は、それぞれの活性の全てのデータの平均値から標準偏差を引いた値よりも小さい場合、活性が強いとした。
Abutilon theophrasti
Aegilops cylindrica
Agropyron repens
Agrostis canina
Agrostis capillaris
Agrostis castellana
Agrostis gigantea
Agrostis stolonifera
Agrostis tenuis
Alopecurus geniculatus
Alopecurus myosuroides
Alopecurus pratensis
Althaea officinalis
Amaranthus albus
Amaranthus lividus
Amaranthus palmeri
Amaranthus rudis
Amaranthus spinosus
Amaranthus thumbergii
Ammi majus
Anisantha madritensis
Anisantha rigida
Anoda cristata
Anthriscus caucalis
Apera spica-venti
Arctium minus
Argemone mexicana
Artemesia vulgaris
Astragalus danicus
Avena fatua
Avena sterilis
Brachiaria decumbens
Bromus arvensis
Bromus hordeaceus
Bromus lanceolatus
Bromus racemosus
Bromus rubens
Bromus secalinus
Bromus sterilis
Camelina sativa
Cardamine pratensis
Cenchrus longispinus
Centaurea maculosa
Centaurea phrygia
Centaurium erythraea
Centaurium tenuiflorum
Centrosema pubescens
Cerastium frontanum
Cerastium glomeratum
Chamaenerion angustifolium
Chloris gayana
Chrysanthemum parthenium
Cichorium intybus
Cnicus benedictus
Cochlearia danica
Cochlearia officinalis
Conium maculatum
Consolida orientalis
Crepis capillaris
Criteson secalinum
Cymbopogon citratus
Cyperus difformis
Cyperus fuscus
Dactylis glomerata
Dactylocenium aegyptum
Datura stramonium
Deschampsia flexuosa
Desmodium ovalifolium
Digitalis purpurea
Digitaria adscendens
Digitaria ischaemum
Diplotaxis muralis
Dipsacus fullonum
Dipsacus pilosus
Dipsacus sylvestris
Echinochloa colonum
Echinochloa crus galli
科　名
SW法
ＤＰ法
PB法
総合評価
ｲﾈ科
ｲﾈ科
ｲﾈ科
ｲﾈ科
ﾑﾗｻｷ科
ﾑﾗｻｷ科
ﾑﾗｻｷ科
ｾﾘ科
ｾﾘ科
ｾﾘ科
ｲﾈ科
ﾀﾃﾞ科
ﾀﾃﾞ科
ﾌｳﾛｿｳ科
ｱﾌﾞﾗﾅ科
ｾﾘ科
ﾄｳﾀﾞｲｸﾞｻ科
ﾄｳﾀﾞｲｸﾞｻ科
ｺﾞﾏﾉﾊｸﾞｻ科
ｲﾈ科
ｲﾈ科
ｶﾔﾂﾘｸﾞｻ科
ｶﾔﾂﾘｸﾞｻ科
ｹｼ科
ｱｶﾈ科
ﾌｳﾛｿｳ科
ﾌｳﾛｿｳ科
ﾌｳﾛｿｳ科
ﾌｳﾛｿｳ科
ﾌｳﾛｿｳ科
ﾊﾞﾗ科
ﾊﾞﾗ科
ｾﾘ科
ｱﾌﾞﾗﾅ科
ｲﾈ科
ﾅｽ科
ｾﾘ科
ｾﾘ科
ﾋﾙｶﾞｵ科
ﾋﾙｶﾞｵ科
ﾋﾙｶﾞｵ科
ｱﾌﾞﾗﾅ科
ｲｸﾞｻ科
ｲｸﾞｻ科
ｺﾞﾏﾉﾊｸﾞｻ科
ﾏﾒ科
ﾏﾒ科
ﾏﾒ科
ｷｷｮｳ科
ｾﾘ科
ｾﾘ科
ｾﾘ科
ｱﾌﾞﾗﾅ科
ﾌﾄﾓﾓ科
ｾﾘ科
ｲﾈ科
ｲﾈ科
ｲﾈ科
ﾏﾒ科
ﾏﾒ科
ﾅﾃﾞｼｺ科
ｱｵｲ科
ｱｵｲ科
ｱｵｲ科
ｱｵｲ科
ｼｿ科
ｾﾘ科
ｾﾘ科
ｾﾘ科
ｾﾘ科
ｾﾘ科
ｾﾘ科
ﾏﾒ科
ﾏﾒ科
ﾅﾃﾞｼｺ科
ﾅﾃﾞｼｺ科
ﾏﾒ科
38
46
25
22
73
28
39
82
96
77
19
60
57
69
39
37
18
80
63
35
61
47
33
27
39
77
57
72
82
56
73
56
37
17
20
14
81
78
25
45
75
42
54
49
15
23
47
36
42
51
69
50
36
68
35
24
29
50
40
68
48
26
24
21
12
26
63
70
32
65
30
50
45
28
58
37
13
109
109
94
131
89
129
107
124
96
107
135
107
92
131
111
97
64
136
97
110
97
127
100
153
100
77
104
117
89
98
115
102
115
112
100
27
101
88
96
84
109
71
88
111
92
110
77
122
115
96
130
74
94
138
113
84
114
118
18
115
99
108
100
48
88
107
119
110
110
108
98
93
88
91
93
109
61
55
68
30
54
41
43
46
79
62
79
22
54
69
21
16
48
17
39
75
61
29
79
85
30
54
21
38
37
39
33
72
74
65
22
46
47
79
69
57
66
72
25
83
40
59
59
43
40
37
72
61
51
38
27
61
56
81
60
61
36
32
54
81
29
34
83
92
57
47
102
61
94
28
39
28
31
29
0
0
1
1
0
0
0
0
0
0
2
0
0
1
1
0
3
0
0
0
0
0
0
0
0
1
0
0
0
0
0
0
0
2
1
2
0
0
1
0
0
2
0
0
1
1
0
0
0
0
0
1
0
1
0
1
0
0
1
0
0
1
1
2
1
1
0
0
0
0
0
0
1
0
1
0
2
学　名
Melilotus officinalis
Melilotus sulcata
Mentha arvensis
Myosotis arvensis
Nicotiana sylvestris
Oenanthe crocata
Oenothera erythrosepala
Onobrychis viciifolia
Ononis spinosa
Panicum clandestinum
Panicum miliaceum
Panicum virgatum
Papaver dubium
Papaver hybridum
Paspalum scrobiculatum
Pennisetum glaucum
Phalaris aquatica
Phalaris canariensis
Phalaris minor
Physalis alkekengi
Picris echioides
Pimpinella anisum
Plantago lanceolata
Poa pratensis
Poa trivialis
Poterium sanguisorba
Prunella vulgaris
Puccinellia distans
Pueraria javanica
Ricinus gibsonii
Rorippa austriaca
Rottboellia exaltata
Rubus fruticosus
Rumex conglomeratus
Rumex crispus
Rumex hydrolapathum
Rumex patientia
Rumex rupestris
Rumex sanguineus
Rumex stenophyllus
Sagina subulata
Salvia hormoroides
Sanguisorba minor
Senecio jacobaea
Senecio vulgaris
Sesbania exaltata
Setaria faberi
Setaria macrostachia
Setaria verticillata
Setaria viridis
Sida alba
Sida rhombifolia
Sida spinosa
Sison ammonum
Sorghum bicolor
Sorghum sudanense
Spergularia bocconei
Sporobolus cryptandrus
Stachys annua
Tanacetum vulgare
Tephrosia purpurea
Trifolium album
Trifolium arvense
Trifolium campestre
Trifolium dubium
Trifolium pratense
Tripleurospermum maritimum
Verbascum nigrum
Verbascum thapsus
Verbena officinalis
Veronica anagalloides
Veronica persica
Vicia hirsuta
Vulpia bromoides
Vulpia myuros
Xanthium spinosum
Xanthium strumarium
和　名
ｾｲﾖｳｴﾋﾞﾗﾊｷﾞ
科　名
ﾏﾒ科
ﾏﾒ科
ｼｿ科
ﾑﾗｻｷ科
ﾅｽ科
ｾﾘ科
ｱｶﾊﾞﾅ科
ﾏﾒ科
ﾊﾘﾓｸｼｭｸ
ﾏﾒ科
ｲﾈ科
ｲﾈ科
ｲﾈ科
ﾅｶﾞﾐﾋﾅｹﾞｼ
ｹｼ科
ﾄｹﾞﾐｹﾞｼ
ｹｼ科
ｲﾈ科
ﾄｳｼﾞﾝﾋﾞｴ
ｲﾈ科
ｵﾆｸｻﾖｼ
ｲﾈ科
ｶﾅﾘｰｸｻﾖｼ
ｲﾈ科
ﾋﾒｶﾅﾘｰｸｻﾖｼ
ｲﾈ科
ﾖｵｼｭﾎｳｽﾞｷ
ﾅｽ科
ﾊﾘｹﾞｺｳｿﾞﾘﾅ
ｾﾘ科
ｱﾆｽ
ｾﾘ科
ﾍﾗｵｵﾊﾞｺ
ｵｵﾊﾞｺ科
ｲﾈ科
ｲﾈ科
ｵｵｽｽﾞﾒﾉｶﾀﾋﾞﾗ
ﾊﾞﾗ科
ｼｿ科
ｱﾚﾁﾀﾁﾄﾞｼﾞｮｳﾂﾅｷﾞ
ｲﾈ科
ﾏﾒ科
アカネ科
ﾐﾐｲﾇｶﾞﾗｼ
ｱﾌﾞﾗﾅ科
ﾂﾉｱｲｱｼ
ｲﾈ科
ﾊﾞﾗ科
ｱﾚﾁｷﾞｼｷﾞｼ
ﾀﾃﾞ科
ﾀﾃﾞ科
ﾀﾃﾞ科
ﾀﾃﾞ科
ﾀﾃﾞ科
ﾀﾃﾞ科
ﾀﾃﾞ科
ﾅﾃﾞｼｺ科
ｼｿ科
ｵﾗﾝﾀﾞﾜﾚﾓｺｳ
ﾊﾞﾗ科
ｾﾘ科
ﾉﾎﾞﾛｷﾞｸ
ｾﾘ科
ﾏﾒ科
ｱｷﾉｴﾉｺﾛｸﾞｻ
ｲﾈ科
ｲﾈ科
ｻﾞﾗﾂｷｴﾉｺﾛｸﾞｻ
ｲﾈ科
ｴﾉｺﾛｸﾞｻ
ｲﾈ科
ｱｵｲ科
ｱｵｲ科
ｱｵｲ科
ｾﾘ科
ｿﾙｶﾞﾑ
ｲﾈ科
ｽｰﾀﾞﾝｸﾞﾗｽ
ｲﾈ科
ｳｼｵﾊﾅﾂﾒｸｻ
ﾅﾃﾞｼｺ科
ｲﾈ科
ｼｿ科
ﾖﾓｷﾞｷﾞｸ
ｾﾘ科
ﾏﾒ科
ﾏﾒ科
ﾏﾒ科
ﾏﾒ科
ｺﾒﾂﾌﾞﾂﾒｸｻ
ﾏﾒ科
ｱｶﾂﾒｸｻ
ﾏﾒ科
ｾﾘ科
ｺﾞﾏﾉﾊｸﾞｻ科
ﾋﾞﾛｰﾄﾞﾓｳｽﾞｲｶ
ｺﾞﾏﾉﾊｸﾞｻ科
ｸﾏﾂﾂﾞﾗ科
ｺﾞﾏﾉﾊｸﾞｻ科
ｺﾞﾏﾉﾊｸﾞｻ科
ｽｽﾞﾒﾉｴﾝﾄﾞｳ
ﾏﾒ科
ｲﾈ科
ﾅｷﾞﾅﾀｶﾞﾔ
ｲﾈ科
ｾﾘ科
ｾﾘ科
SW法
ＤＰ法
PB法
総合評価
2
1
32
64
71
25
65
64
45
39
58
45
44
17
40
36
49
50
26
60
54
25
76
53
32
63
76
15
38
45
16
63
24
73
72
79
84
76
71
55
31
59
70
42
37
16
24
25
28
19
9
35
58
72
21
38
36
51
32
38
22
29
51
59
52
56
59
72
69
77
52
39
29
46
18
21
42
62
37
75
88
100
95
113
105
68
95
81
104
98
104
112
113
42
94
86
88
109
100
101
122
119
89
86
20
122
102
78
108
90
92
147
95
96
103
128
123
81
103
83
141
103
99
124
99
71
86
114
100
121
114
80
30
99
122
106
80
133
110
121
119
82
110
98
126
101
87
148
117
77
81
81
104
120
19
30
97
37
40
93
47
49
32
39
31
62
25
17
65
30
26
34
39
40
46
56
75
55
34
47
91
64
33
75
53
35
67
85
70
75
60
78
82
70
53
23
70
43
77
45
23
37
29
26
51
71
46
56
21
18
44
30
54
93
35
15
48
42
21
65
70
43
24
82
97
46
3
46
30
64
79
3
2
1
0
0
1
0
0
1
0
0
0
1
2
0
0
2
0
1
0
0
1
0
0
0
0
0
2
0
0
1
0
1
0
0
0
0
0
0
0
0
1
0
0
0
1
2
1
1
2
1
0
0
0
2
2
0
0
0
0
1
1
0
0
1
0
0
0
1
0
0
0
1
0
1
1
0
Evaluation of Allelopathy by specific bioassays
and evaluated about 4,000 species in 20 years

1) Medicinal Plants
2) Tropical Plants
are Allelopathic
3) Endangered Plants
4) Invasive Alien Plants
5) Oats, Barley, Wheat
(My dream) Database for Allelopathic activity evaluated by PB, SW, DP method
16
2) Isolation of
Allelochemicals
based on
Total Activity
17
Chromatography
(TLC, HPLC …)
HO
OH
H
S
S
H
O
Zeylanoxide A
試料添加
薄層クロマトグラフィー用
シリカゲルプレート
C
さらに展開
溶媒（油）で展開
さらに展開
良好な分離！
H
H
H
H
HO
C
OH
H
S
S
H
O
epi-Zeylanoxide A
Structure
GC-MS, MS,
NMR, IR, ESR,
X-ray analysis,
etc
Solvent Extraction
Conventional Method for Isolation of natural chemicals
18
New strategies for isolation of
allelochemicals in agriculture
Comparison of Total Activity and Specific Activity
Target
Compounds
Direct use
Allelochemicals
of Allelopathy in action in situ
Index (g-weight base)
Total Activity
= 1/EC50×Concentration in Plants
1/（kg/L） x (kg/L in fresh weight)
=(L/kg) x (kg/L) = (no unit)
high value ～ high activity
Use of Natural Bioactive
Chemicals
Chemicals with
high specific
activity
Specific Activity
= EC50 (50% inhibitory Concentration)
(kg/L)～ppm
low ppm ～ high activity
19
Isolation of L-DOPA from Velvet bean
Endemic plant
to Himalayas
Distributed in
Sino-Japan Area
Used as food
in Japan from
more than
1000 years ago
Total activity (but now
disappearing)
= 200
Fujii Y, Shibuya T. and Yasuda T. L-3,4-dihydroxyphenylalanine as an allelochemical candidate from
Mucuna pruriens (L.) DC. var. utilis. Agricultural and Biological Chemistry 55, 617-618. (1991)
20
Isolation of cyanamide from hairy vetch
Cyanamide was
already known as
synthetic fertilizer,
but never isolated as
natural chemical
Endemic plant
to central Asia,
now we are
recommending
to Japanese
farmers for
weed control
Total activity
= 35 - 100
Kamo T, Hiradate S. and Fujii Y. First isolation of natural cyanamide as a possible
allelochemical from hairy vetch Vicia villosa.，J. Chem. Ecol.，29 (2), 275-284 (2003)
21
3) Practical use of
Allelopathic
Cover Crops
22
Mucuna (velvetbean)
(Mucuna pruriens)
for Green manure, Food, Medicinal
plants, and Cover Crop
Fujii, Y. Allelopathy of Velvetbean: Determination and Identification of
Allelopathic Substances. H. G. Cutler and S. Cutler (Eds), Biologically Active
Natural Products: Agrochemicals, CRC Press, pp.33-47 (1999)
23
Ｖｅｌｖｅｔbean : Mucuna pruriens var. utilis
High production of forage
,
Legume, rich in protein
24
Table 1. Assessment of Allelopathic Activity of
Leguminous Cover Plants by Plant Box Method
Scientific name (English name)
Mucuna pruriens var.utilis (Mucuna)
Vicia faba (Broad Bean)
Vicia villosa (Hairy Vetch)
Calopogonium mucunoides
Melilotus albus (White Sweet Clover)
Vicia sativa (Common Vetch)
Medicago rugosa
Canavalia ensiformis (Jack Bean)
Pueraria phaseoloides (Tropical Kudzu)
Vigna angularis (Azuki bean)
Medicago sativa (Alfalfa)
Trifolium incarnatum (Crimson Clover)
Tephrosia candida (White Tephrosia)
Cajanus cajan (Pigeon Pea)
Latyrus sativus (Grass Pea)
Cicer arietinum (Chickpea)
Vigna radiata (Mung Bean)
Stylosanthes hamata (Stylo)
Arachis hypogaea (Peanut)
Trifolium pratense (Red Clover)
Crotaralia juncea (Sunn Hemp)
Astragalus sinicus (Chinese Milk Vetch)
Lupinus albus (White Lupine)
Trifolium subterraneum (Sub Clover)
Trifolium repens (White Clover)
Glycine max (Soybean)
[%]*1
87
81
80
78
77
75
74
72
71
69
68
64
63
62
59
56
53
52
51
47
43
41
40
30
28
24
Criteria*2
★★★
★★
★★
★★
★★
★★
★★
★★
★★
★
★
★
★
★
★
★
Plant Box Method
(for root exudates)
*1 Inhibition (%) means allelopathic activity
measured by the Plant Box Test.
All data are compared to the control and
100 means complete inhibition.
*2 Criteria for the allelopathic activity are ;
★★★ ; >85 %, ★★ ; 70 to 84% ,
★ :<69 %, respectively.
25
Table
Weed population in continuous cropping fields
──────────────────────────────────────
Crop
Treatment
Weed population
Weed species observed
6)
(g dry weight per m2)
──────────────────────────────────────
Upland Rice
3yr.c 1)
5.11(49.4) 4) 1), 3), 5), 6), 7), 8), 9), 10), 11) 5)
Egg plant
3yr.c
16.82(40.1)
1), 2), 3), 5), 6), 7), 8), 9), 10), 11), 12),
Tomato
3yr.c
4.92(64.9)
1), 5), 6), 9), 12), 13), 17)
Velvetbean
2yr.c
0.00( 0.0)
no emergence
Fallow
3yr.f
3)
0.97(37.3)
13), 14)
1), 2), 6), 10), 12), 13), 15), 16)
──────────────────────────────────────
1) Continuous cropping for 3 years.
After Velvetbean,
little weed !
2) Cultivated for 1 year, followed by fallow next year(test year).
3) Fallow for 3 years, without fertilizer.
4) Numbers in parenthesis are percentages of chickweed, a dominat species.
5) Species appeared in each plot: 1) Sticky chickweed (Cerastium glomeratum),
2) 'Miminagusa' (Cerastium vulgatum var. augustifolium),
3) Annual fleabane
(Erigeron annuus), 4) Philadelphia fleabane (Erigeron philadelphicus),
5) Starwort (Stellaria alsine var. undulata),
geniculatus),
6) Floating foxtail (Alopecurus
7) Narrowleaf vetch (Vicia angustifolia),
(Cardamine flexuosa),
8) Flexuosa bittercress
9) 'Inugarashi' (Rorippa atrovirens), 10) Common dandelion
(Taraxacum officinale), 11) Japanese mugwort (Artemisia princeps), 12) Danadian
fleabane (Erigeron canadensis), 13) 'Hahakogusa' (Gnaphalium affine), 14) Blady
grass (Imperata cylindrica), 15) Meadowgrass (Poa annua), 16) Creeping woodsorrel (Oxalis corniculata), 17) Shepherd's-purse (Capsella bursa-pastoris),
18) Prickly sowthistle (Sonchus asper).
Fujii, Y. Shibuya, T. and Usami, Y. Allelopathic effect of Mucuna pruriens on the
Source: Fujii, Y. Shibuya, T. and Usami, Y. (1991): Allelopathic effect of Mucuna pruriens
appearance
ofof weeds.
weeds,
Weed
on the appearance
Weed Res.
36, 43-49 Res, Japan 36, 43-49 (1991)
26
Allelochemical in Mucuna (Velvetbean)
L-3,4-Dihydroxyphenylalanine (L-DOPA)
HO
HO


COOH
NH2
Extremely high concentration in Mucuna pruriens leaves,
roots (0.5 to 1.5%) and seeds (5 to 8%) in fresh wt.
Relatively strong inhibition to broadleaf weeds
(EC50=5-50ppm), but not so active to grass (monocot.)
Fujii, Y., T. Shibuya, and T. Yasuda, L-3,4-Dihydroxyphenylalanine as an
allelochemical candidate from Mucuna pruriens(L.) DC.var.utilis.
Agric.Biol.Chem. 55(2), 617-618 (1991)
27
How to use Mucuna
1) Green Manure
2) Vegetable (Young pod)
3) Seeds as food, but after
proper Cooking.
4) Medicine, as a source of
L-DOPA
28
Hairy vetch
(Vicia villosa)
for Green manure and Weed control
Fujii, : Screening and Future Exploitation of Allelopathic Plants as Alternative
Herbicides with Special Reference to Hairy Vetch. R. K. Kohli, H. P. Singh, and D.
R. Batish (Eds), Allelopathy in Agroecosystems, pp.257-275 (2001)
29
First observation of strong weed suppression on the field, August, 1990
30
Table
Field
Effect of Cover Crops on Weed Control in Abandoned Paddy
; First Trial on the Experimental Station (1992-1993)
Cover Crop (English Name) First Sampling (May 7, 1993)
Control(No-weeding)
Astragalus sinicus（Chinese Milk Vetch)
Vicia villosa (Hairy Vetch)
A. sativa＋V. villosa（Oat and Hairy Vetch Mix.)
Second Sampling (June 10, 1993)
Control (No-weeding)
Astragalus sinicus（Chinese Milk Vetch)
Vicia villosa (Hairy Vetch)
A. sativa＋V. villosa（Oat and Hairy Vetch mix.)
W
(%)*1
0 a *2
82 b
99 b
99 b
0a
59 b
100 c
100 c
Crop Yield
[g m-2]
---431 a
584 ab
730 b
---135 a
147 b
137 a
*1 Percentage of weed control. 100 % means complete control. Dry weight
of weeds in the control plot are 281 (First) and 155 (Second) g m-2.
Fujii, Y. Screening and future exploitation of allelopathic plants as alternative
herbicides with special reference to hairy vetch，J. Crop Prod.，4 (2) , 257-275 (2001)
31
Recommendation
of Hairy Vetch
In Newspaper,
1994
(Japan Agricultural
News)
32
Orange Farm
Mr.Ochi
（Ehime Pref.)
１９９１年
33
34
34
Identification of Cyanamide as
Allelochemical from Hairy vetch
★ Cyanamide H
N C N
H
★ known as active constituents of
synthetic fertilizer (Calcium
cyanamide) First finding from plants
Kamo, T.,
Hiradate,
S. andproducs)
Fujii, Y.: First isolation of natural cyanamide as
(as
natural
a possible allelochemical from hairy vetch Vicia villosa . J. Chemical Ecol.
29 (2), 275-283 (2003)
35
36
Hairy Vetch used as
cover crop in Japan
more than10,000 ha
(estimated from 300 tons seed)
No.2 as cover crop
…after 18 years
37
Other
Promising
Allelopathic
Cover Plants
38
39
Ground Cover Plants
with Allelopathic activity
Perfume, Medicinal chemicals
Resistant for insects,
disease & weeds
Honey
Beautiful flowers
O2 ↑
Useful
as
Forage
Prevent
Erosion
Suppress weeds
N2 fixation
Food (Fruit,
Seed, Tuber...)
Clean polluted soil by
toxic chemicals
Cultivate
soil by root
system
Keep soil
moisture
Green manure by
Nitrogen fixation and
organic carbon
Resistant for nematoda,
and soil disease
Phyto-Lark : Ground Cover plants with allelopathy
40
Three key messages
• Specific bioassays for Allelopathy, and
evaluated about 4,000 plants.
• New Allelochemicals Isolated by new
methodology, “Total Activity Method”
• Ground Cover Plants with allelopathiy
(= Phytolark ) provides natural and
environmentally friendly way of managing
weeds without herbicides.
41
Questions?
Contact details
[email protected]
42

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