椙山女学園大学研究論集 第 47 号（自然科学篇）2016

Bioactivity and Nutrient Content of Wild Edible Plants

Tamio MASE*, Sachiyo SHITASUE*, Kanae UCHIDA*,

Shiori UOZUMI* and Shinobu ISSHIKI*

Abstract

The nutritional content and bioactivity of 27 kinds of wild edible plants collected
from their natural environment in early spring were investigated. The highest
levels of ascorbic acid, polyphenol, and soluble protein in the plant samples were
940, 696, and 8.29 mg 100 g−1, respectively. Polyphenols were abundant in all
species. Portulaca oleracea, which is widespread during the summer, showed the
highest hyaluronidase inhibitory activity suggesting anti-allergic properties. All
showed antioxidant bioactivity, defined as DPPH radical-scavenging activity, and
angiotensin-converting enzyme (ACE, EC 3.4.15.1) inhibitory activity, suggesting
a role in the prevention of hypertension. These wild edible plants are therefore
expected to be not only nutritious but also of medicinal value.

Introduction

There are many kinds of edible wild plants known in Japan as sansai, along with many others
not widely eaten but of potential nutritional and medicinal value. Various plant components such as
polyphenols and dietary fiber have attracted substantial attention from the viewpoint of functional
health; therefore, understanding the nutritional components and health functionality of these
lesser known edible plants would be beneficial. Such plants could provide dietary factors such
as minerals, proteins, vitamin C, and polyphenols (Shitasue et al., 2015) as well as medicinally
valuable anti-bacterial, hepatoprotective, and anticarcinogenic effects.
Yildirim et al. (2001) have reported the nutrition content of wild plants used as vegetables in
the Upper Coruh Valley, Turkey, while Romojaro et al. (2013) have reported the health benefits of
various wild edible plants. Against this background, the aim of the present study was to investigate
the nutritional content and bioactivity of lesser-known wild edible plants in Japan.

* School of Life Studies, Department of Human Nutrition

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Materials and Methods

Experimental materials The nutritional value and bioactivity of 27 wild edible plants growing
in their natural environment around Ichinomiya City, Japan, were collected in early spring. Table 1
shows the taxonomic identifications of these plants defined in accordance with Hashimoto (2007).

Table 1. The wild edible plants included in the analysis.

Binomial name Family Japanese common name

Trifolium pratense Fabaceae Akatsumekusa
Vicia sativa subsp. nigra Fabaceae Karasunoendou
Glycyrrhiza glebra Fabaceae Kanzou
Vicia hirsuta Fabaceae Suzumenoendou
Trifolium repens Fabaceae Shirotsumekusa
Pueraria lobate (Wild) Ohwi Fabaceae Kuzu
Rumex acetosa Polygonaceae Suiba
Rumex japonicas Polygonaceae Gishigishi
Rumex conglomeratus Murr Polygonaceae Arechigishigishi
Commeline communis Commelinaceae Tsuyukusa
Plantago lanceolata Plantaginaceae Heraoobako
Youngia japonica Asteraceae Onitabirako
Artemisia indica var. maximowiczii Asteraceae Yomogi
Cirsium japonicum Asteraceae Azami
Picris hieracioides Asteraceae Kouzorina
Erigeron annuus Asteraceae Himejion
Sonchus oleraceus Asteraceae Harunogeshi
Sonchus asper Asteraceae Oninogeshi
Cerastium holosteoides Caryophyllaceae Miminagusa
Lamium amplexicaule Lamiaceae Hotokenoza
Lamium purpurureum Lamiaceae Himeodorikosou
Lamium album Lamiaceae Odorikosou
Portulaca oleracea Portulacaceae Suberihiyu
Oxalis corniculata Oxalidaceae Katabami
Fatoua villosa Moraceae Kuwakusa
Cayratia japoica Vitaceae Yabukarashi
Trigonotis peduncularis Boraginaceae Kyuurigusa

Preparation of plant extracts All plant samples were frozen in a freezer at −85°C then freeze
dried. The freeze-dried samples were then milled into flour using a blender (Wonder Blender WB-
1; Osaka Chemical Ltd., Amagasaki, Japan) and stored at −85°C until use within 3 days. For the
assay, 2.5 g of freeze-dried flour was mixed with 50 mL of distilled water then extracted with a
homogenizer (Homojinizer; IKA Japan Co., Ltd., Higashiosaka, Japan). The resulting mixture was
centrifuged at 10,000 rpm for 10 min and the resulting supernatant solution used for analysis of
plant components and bioactivity.

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Componential analysis The moisture, ascorbic acid, and polyphenol contents were assayed
according to Mase et al. (2012), and the soluble protein content was measured with Bio-Rad protein
reagent (Bio-Rad Laboratories, Berkeley, CA) using bovine serum albumin as the standard.
DPPH radical-scavenging activity DPPH radical-scavenging activity was assayed according
to Mase et al. (2012).
Angiotenshin-converting enzyme inhibitory activity The inhibitory activity of Angiotenshin-
converting enzyme (ACE) was assayed using an ACE Kit WST (Wako Pure Chemical Industries
Ltd., Osaka, Japan).
Cysteine protease inhibitory activity Azocasein (SIGMA) was used as a substrate for cysteine
protease inhibitory activity assay. A total of 1.5 mL of 0.8% substrate solution dissolved in 100
mM Tris-HCl buffer solution (pH 7.0), 0.4 mL of 0.02% cysteine protease, Papain (Sigma), and 0.1
mL of distilled water were included in the reaction mixture. After reaction at 37°C for 30 min, 2
mL of 10% TCA solution was added to terminate the reaction. The mixture was then filtered with
filter paper No.131 (Toyoroshi Co. Ltd., Japan), and the absorbance at 400 nm measured. Inhibitory
activity was defined according to the relative activity of the plant extract solution relative to that of
distilled water.
Hyaluronidase inhibitory activity The assay method was as shown previously (Mase et al.,
2013).

Results and Discussion

The nutritional contents of the 27 wild edible plants are shown in Table 2. The moisture content
of all freeze dried plants was in the range of 4.5–6.4 %. The ascorbic acid contents differed among
the species tested, ranging from 940 mg 100 g−1 in Vicia hirsuta to 67 mg 100 g−1 in Fatoua villosa.
The ascorbic acid contents tended to be rich compared to commonly consumed vegetables such as
Spinacia oleracea (263 mg 100g−1), and Allium fistulosum (170 mg 100g−1) shown in the report
of Shitasue et al. (2015). Ascorbic acid is a water-soluble vitamin (vitamin C), required for the
growth and repair of a wide range of body tissues. It is also an antioxidant, responsible for blocking
damage caused by free radicals (Yamada et al., 2003).
The soluble protein contents were quite varied depending on the plant species tested. In six of
the species, soluble protein could not be detected, whereas in Lamium album (5.10 g 100 g−1), and
Trifolum pratense (8.28 g 100 g−1) contents were high (Table 2). It is possible that the assay method
employed in this study was not sensitive enough for detection of amino acids or low molecular
weight peptides. With regard to polyphenol content, almost all of the samples showed a high
content (254 to 696 mg 100 g−1DW).
Antioxdant activity is shown in Table 2 as values of DPPH radical-scavenging activity. All plants
tested showed DPPH radical-scavenging activity, with Rumex conglomeratus Murr (3.74 mmol
100 g−1DW), Lamium purpurureum (3.41 mmol 100 g−1DW), and Erigeron annuus (3.41 mmol
100 g−1DW) having particularly high activity. Takahashi et al. (2007) and Takahashi et al. (2009)
reported a correlation between DPPH radical-scavenging activity and polyphenol content in local

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Table 2. Polyphenol, ascorbic acid and soluble protein contents, and DPPH radical-scavenging activity.

Binomial name Polyphenols Ascorbic acid Soluble DPPH radical-

protein scavenging
(mg/100 g DW) (mg/100 g DW) (mg/100 g DW) activity
(mmol/100 g DW)
Trifolium pratense 464 717 8.29 0.64
Vicia sativa subsp. nigra 632 717 2.74 1.49
Glycyrrhiza glebra 553 661 1.57 1.79
Vicia hirsuta 289 940 0.85 1.81
Trifolium repens 561 585 1.08 2.51
Pueraria lobate (Wild) Ohwi 570 174 2.61 3.38
Rumex acetosa 696 701 trace 2.53
Rumex japonicas 472 190 trace 1.82
Rumex conglomeratus Murr 647 340 trace 3.74
Commeline communis 358 375 1.09 0.81
Plantago lanceolata 304 367 trace 0.50
Youngia japonica 612 581 0.94 1.96
Artemisia indica var. maximowiczii 254 283 0.64 0.58
Cirsium japonicum 466 437 3.82 1.36
Picris hieracioides 383 395 0.26 0.90
Erigeron annuus 637 279 trace 3.41
Sonchus oleraceus 471 479 4.81 0.58
Sonchus asper 396 210 1.08 0.65
Cerastium holosteoides 260 412 2.13 0.60
Lamium amplexicaule 378 465 1.89 0.80
Lamium purpurureum 645 333 1.15 3.41
Lamium album 659 502 5.10 2.85
Portulaca oleracea 626 188 0.10 1.59
Oxalis corniculata 306 157 trace 0.97
Fatoua villosa 290 67 1.26 1.63
Cayratia japoica 660 610 2.94 0.78
Trigonotis peduncularis 543 709 0.77 2.38

plants; however, in this study, there was no correlation.

All plants tested showed ACE inhibition (Table 3). ACE inhibitor prevents the generation of
angiotensin II from angiotensin I. Angiotensin is an oligopeptide that plays an important role in
the renin-angiotensin system. Angiotensin I is formed from angiotensinogen via the action of
renin, and converted to angiotensin II by ACE. Angiotensin II increases blood pressure and causes
hypertension, and therefore, ACE inhibition is a major target of hypertension control. There are
many reports of ACE inhibition using plant components such as peptides, polyphenols (Loizzo
et al., 2007) and flavonoids (Loizzo et al., 2009). Artemisia indica var maximowicrii, Erigeron
annuns, and Sonchus asper showed high ACE inhibition activity, but activity had no correlation
with polyphenol or soluble protein content.

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Table 3. ACE, cysteine protease, and hyaluronidase inhibition.

Binomial name ACE inhibition Cysteine Hyaluronidase

IC50(mg/mL) protease inhibition (%)
inhibition (%)
Trifolium pratense 5.60 ND 54.0
Vicia sativa subsp. nigra 3.06 4.86 52.7
Glycyrrhiza glebra 0.41 ND ND
Vicia hirsuta 1.08 1.41 8.7
Trifolium repens 3.64 ND ND
Pueraria lobate (Wild) Ohwi 2.48 ND 23.5
Rumex acetosa 2.43 4.81 ND
Rumex japonicas 1.76 ND 2.8
Rumex conglomeratus Murr 7.18 12.87 1.0
Commeline communis 7.93 ND 21.1
Plantago lanceolata 1.31 0.62 46.8
Youngia japonica 3.38 ND 7.5
Artemisia indica var. maximowiczii 31.05 ND 50.1
Cirsium japonicum 4.26 ND 39.6
Picris hieracioides 1.93 ND 4.8
Erigeron annuus 16.18 ND 49.4
Sonchus oleraceus 8.62 8.72 41.1
Sonchus asper 15.15 2.42 ND
Cerastium holosteoides 6.36 ND 63.1
Lamium amplexicaule 5.50 1.86 7.1
Lamium purpurureum 3.69 ND 45.5
Lamium album 2.73 ND ND
Portulaca oleracea 4.73 9.63 68.5
Oxalis corniculata 5.27 3.17 ND
Fatoua villosa 10.65 3.60 31.0
Cayratia japoica 9.78 7.39 40.7
Trigonotis peduncularis 3.11 ND ND
ND: not detected.

The cysteine protease inhibition activities of the plant samples are shown Table 3. Cathepsin K
(EC 3.4.22.38), a cysteine protease, plays a role in degradation of the bone matrix via osteoclasts.
Degradation of the bone matrix subsequently causes osteoporosis, which is a serious disease among
women. Cathepsin K inhibition is thus known to be useful in treatment of osteoporosis (Boonen et
al., 2012; Miyazaki and Sawada, 2013). In this study, we employed papain as the cysteine protease
instead of cathepsin K. Of the plants tested, 12 showed inhibitory activity, and of these, Rumex
conglomeratus Murr showed the highest activity (12.87 %).
The hyaluronidase inhibitory activity of the plant samples was also tested. Hyaluronidase
mediates inflammation via histamine release from mast cells. Hyaluronidase inhibitors are
therefore thought to be effective in suppressing allergies and inflammation (Furusawa et al., 2011).

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Twenty of the tested plant species showed inhibitory activity, with Portulaca oleracea (68.5 %)
showing particularly high activity. Portulaca oleracea was furthermore found to contain many
sticky polysaccharides such as hyaluronic acid, as a substrate of hyaluronidase. These sticky
polysaccharides seem to work as an antagonist of hyaluronic acid.

Conclusions

The results of this investigation of 27 wild edible plants revealed high nutritional properties
such as polyphenol and ascorbic acid content as well as strong ACE, cysteine protease, and
hyaluronidase inhibitory activity. These findings suggest that these species could be used not only
as alternatives to conventionally consumed vegetables, but also in the prevention or treatment of
allergies, osteoporosis, and hypertension (Romjaro et al., 2013)

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