This article attempts to trace the origin of tea. The author believes the ancient Chinese tea, "chia", is either Jicha (water extract from the pith of Acacia catechu that grows naturally in the mountainous border between the Yunnan province of China and southern Asian countries) or Jicha-Kagikazura (water extract from the young branches and leaves of Uncaris gambir, originally found in India/Sri Lanka). Both were pulverized after being kiln-dried and then mixed with water to produce a thick suspension, or tea. Although the drink is bitter and has an astringent property, it has a particular flavor with a refreshing after-taste. Its components with medicinal properties include tannin, catechin, and various flavonoids, making us believe it was worthwhile for the people at the time to consume the drink regularly. Generally speaking, tea cultivation in China flourished south of the Yangzi Jiang River including the present Zhejiang and Anhui provinces. Depending on the regions, there were words for tea in various languages, including the names of places where particular teas were grown. In addition to the names that appear in the famous Chajing book, it is interesting to note Da Fang pronounced tea as "TAH". Because the area south of the Yangzi Jiang has traditionally been active in foreign trade since the ancient and middle ages. People in this region consumed various foreign originated teas as well. This included Gambir, which was introduced to southern Asia (including present Malaysia and Indonesia) and was consumed as an herbal tea under names such as Guo Luo or Ju Luo teas. Paan, from India, also uses Gambir paste and was a popular chewing refreshment to prevent diseases caused by miasma as well as to keep one's mouth clean. The name A-sen-yaku used in Japan was taken from the plant name Acasia, and Gambir was used to dye Buddhist monks' Ke-Ra bags to a blackish yellow color. The Daikanwa dictionary states the Ra in the name, which means thin silk, was later replaced with "A". The official name for Ji-cha [Er Cha] in modern China is "Gaiji-cha", [Hal-Er Cha], which comes from the name of a variety of tea made by the Ai-Ni tribal subgroup of the ethnic Ha-Ni in Yunnan province. The [see character in text] character is pronounced "ni", which is a homophony of [character in text]. Based on these facts, "Ai-Ni" should be considered the same as "Hai-Ni". Because the ethnic groups in Yunnan province used primitive and tough tea leaves, which were eaten instead of being infused in water, the leaves were first fermented by being buried in the ground. Even today, people of these ethnic groups prefer fungus-fermented black tea with a particular flavor. In contrast, the ethnic Hans used and still use improved and softer young shoots of tea leaves to prepare mainly green tea. It has recently been discovered that Acapsia, as well as Gambir, has anti-oxidant properties, and that consumption over time is effective against many lifestyle-related adult diseases. It may be well worthwhile to cast fresh light upon ancient tea drinking customs.
This article presents the abilities and efficiencies of five different strains of Agrobacterium rhizogenes (strain ATCC 31798, ATCC 43057, AR12, A4 and A13) to induce hairy roots on Solanum mammosum through genetic transformation. There is significant difference in the transformation efficiency (average number of days of hairy root induction) and transformation frequency for all strains of A. rhizogenes (P < 0.05). Both A. rhizogenes strain AR12 and A13 were able to induce hairy root at 6 days of co-cultivation, which were the fastest among those tested. However, the transformation frequencies of all five strains were below 30 %, with A. rhizogenes strain A4 and A13 showing the highest, which were 21.41 ± 10.60 % and 21.43 ± 8.13 % respectively. Subsequently, the cultures for five different hairy root lines generated by five different strains of bacteria were established. However, different hairy root lines showed different growth index under the same culture condition, with the hairy root lines induced by A. rhizogenes strain ATCC 31798 exhibited largest increase in fresh biomass at 45 days of culture under 16 h light/8 h dark photoperiod in half-strength MS medium. The slowest growing hairy root line, which was previously induced by A. rhizogenes strain A13, when cultured in optimized half-strength MS medium containing 1.5 times the standard amount of ammonium nitrate and potassium nitrate and 5 % (w/v) sucrose, had exhibited improvement in growth index, that is, the fresh biomass was almost double as compared to its initial growth in unmodified half-strength MS medium.
Ganoderma boninense, the main causal agent of oil palm (Elaeis guineensis) basal stem rot (BSR), severely reduces oil palm yields around the world. To reduce reliance on fungicide applications to control BSR, we are investigating the efficacy of alternative control methods, such as the application of biological control agents. In this study, we used four Streptomyces-like actinomycetes (isolates AGA43, AGA48, AGA347 and AGA506) that had been isolated from the oil palm rhizosphere and screened for antagonism towards G. boninense in a previous study. The aim of this study was to characterize these four isolates and then to assess their ability to suppress BSR in oil palm seedlings when applied individually to the soil in a vermiculite powder formulation. Analysis of partial 16S rRNA gene sequences (512 bp) revealed that the isolates exhibited a very high level of sequence similarity (> 98%) with GenBank reference sequences. Isolates AGA347 and AGA506 showed 99% similarity with Streptomyces hygroscopicus subsp. hygroscopicus and Streptomyces ahygroscopicus, respectively. Isolates AGA43 and AGA48 also belonged to the Streptomyces genus. The most effective formulation, AGA347, reduced BSR in seedlings by 73.1%. Formulations using the known antifungal producer Streptomyces noursei, AGA043, AGA048 or AGA506 reduced BSR by 47.4, 30.1, 54.8 and 44.1%, respectively. This glasshouse trial indicates that these Streptomyces spp. show promise as potential biological control agents against Ganoderma in oil palm. Further investigations are needed to determine the mechanism of antagonism and to increase the shelf life of Streptomyces formulations.
The habitats of Eurycoma longifolia Jack, a slender tree, are jungles in Malaysia and Indonesia. It belongs to the family Simaroubaceae and is a source of quassinoids with anabolic, antimalarial and cytostatic activity. In this study, conducted during 2008 in Mae Sot, Thailand, a standardized extract of E. longifolia containing three major quassinoids, eurycomanone (1), 13,21-dihydroeurycomanone (2) and 13alpha(21)-epoxyeurycomanone (3) was evaluated for antiplasmodial activity against Plasmodium falciparum and its activity has been compared with that of artemisinin, using 38 fresh parasite isolates and assessment of inhibition of schizont maturation. The IC(50), IC(90) and IC(99) values for artemisinin were 4.30, 45.48 and 310.97 microg/l, and those for the root extract from E. longifolia 14.72, 139.65 and 874.15 microg/l respectively. The GMCOC for artemisinin was 337.81 mug/l, and for the plant extract it was 807.41 microg/l. The log-concentration probit regressions were parallel. The inhibitory activity of the E. longifolia extract was higher than that expected from the three quassinoids isolated from the plant, suggesting synergism between the quassinoids or the presence of other unidentified compounds.
In this study, Centella asiatica and surface soils were collected from 12 sampling sites in Peninsular Malaysia, and the barium (Ba) concentrations were determined. The Ba concentration [μg/g dry weight (dw)] was 63.72 to 382.01 μg/g in soils while in C. asiatica, Ba concentrations ranged from 5.05 to 21.88 μg/g for roots, 3.31 to 11.22 μg/g for leaves and 2.37 to 6.14 μg/g for stems. In C. asiatica, Ba accumulation was found to be the highest in roots followed by leaves and stems. The correlation coefficients (r) of Ba between plants and soils were found to be significantly positively correlated, with the highest correlation being between roots-soils (r=0.922, p<005), followed by leaves-soils (r=0.890, p<005) and stems-soils (r=0.848, p<005). This indicates that these three parts of C. asiatica are good biomonitors of Ba pollution. For the transplantation study, four sites were selected as unpolluted [(Universiti Putra Malaysia (UPM)], semi-polluted (Seri Kembangan and Balakong) and polluted sites (Juru). Based on the transplantation study under experimental field and laboratory conditions, Ba concentrations in C. asiatica were significantly (p<0.05) higher after three weeks of exposure at Seri Kembangan, Balakong and Juru. Thus, these experimental findings confirm that the leaves, stems and roots of C. asiatica can reflect the Ba levels in the soils where this plant is found. Three weeks after back transplantation to clean soils, the Ba levels in C. asiatica were still higher than the initial Ba level even though Ba elimination occurred. In conclusion, the leaves, stems and roots of C. asiatica are good biomonitors of Ba pollution.
Fungal endophytes are found inside host plants but do not produce any noticeable disease symptoms in their host. In the present study, endophytic Fusarium species were isolated from roots of lawn grass (Axonopus compressus). A total of 51 isolates were recovered from 100 root segments. Two Fusarium species, F. oxysporum (53%) and F. solani (47%), were identified based on macroconidia and conidiogenous cell morphology. The detection of endophytic F. oxysporum and F. solani in the roots of lawn grass contributes to the knowledge of both the distribution of the two Fusarium species and the importance of roots as endophytic niches for Fusarium species.
Experiments were carried out in the laboratory and greenhouse to determine the growth inhibitory effects of Grassohopper’s cyperus (Cyperus iria L.) on the seedlings of 5 Malaysian rice varieties namely MR211, MRQ74, MR220, MR84 and MR232. Three concentrations of the aqueous extract of the weed (12.5, 25.0 and 50.0 g/l) and weed debris (5, 10 and 20 g dry debris/1000 g soil) were used to test the allelopathic effect of
C. iria on the growth of the rice plants. The weed leaf, stem and root extracts reduced the growth of the rice seedlings and showed selective activity in the varieties. The C. iria leaf and stem extracts showed comparatively higher growth inhibitory effects than those from the root. The weed extract caused more reduction in the root length of the rice plant compared to the shoot length. Among the rice varieties tested, MR232 was found to be more susceptible to the weed inhibitory effect. The leaf extract of C. iria at full strength caused root and shoot reduction of MR232 by 88.1% and 73.1% respectively (compared to the control). In most cases the fresh weight of the rice seedlings were more affected than the plant height. Weed debris caused significant reduction of leaf chlorophyll content in all the rice varieties tested with the exception of MR211. The chlorophyll content of MR232 was greatly affected by the weed debris which caused reduction of 36.4% compared to the control. The inhibitory effects of weed extracts and debris on rice growth
parameters were found to be concentration dependent.
Experiments were carried out in the laboratory and greenhouse to determine the growth inhibitory effects of Grassohopper's cyperus (Cyperus iria L.) on the seedlings of 5 Malaysian rice varieties namely MR211, MRQ74, MR220, MR84 and MR232. Three concentrations of the aqueous extract of the weed (12.5, 25.0 and 50.0 g/l) and weed debris (5, 10 and 20 g dry debris/1000 g soil) were used to test the allelopathic effect of C. iria on the growth of the rice plants. The weed leaf, stem and root extracts reduced the growth of the rice seedlings and showed selective activity in the varieties. The C. iria leaf and stem extracts showed comparatively higher growth inhibitory effects than those from the root. The weed extract caused more reduction in the root length of the rice plant compared to the shoot length. Among the rice varieties tested, MR232 was found to be more susceptible to the weed inhibitory effect. The leaf extract of C. iria at full strength caused root and shoot reduction of MR232 by 88.1% and 73.1% respectively (compared to the control). In most cases the fresh weight of the rice seedlings were more affected than the plant height. Weed debris caused significant reduction of leaf chlorophyll content in all the rice varieties tested with the exception of MR211. The chlorophyll content of MR232 was greatly affected by the weed debris which caused reduction of 36.4% compared to the control. The inhibitory effects of weed extracts and debris on rice growth parameters were found to be concentration dependent.
In this study, Centella asiatica and surface soils were collected from 12 sampling sites in Peninsular Malaysia, and the barium (Ba) concentrations were determined. The Ba concentration [µg/g dry weight (dw)] was 63.72 to 382.01 µg/g in soils while in C. asiatica, Ba concentrations ranged from 5.05 to 21.88 µg/g for roots, 3.31 to 11.22 µg/g for leaves and 2.37 to 6.14 µg/g for stems. In C. asiatica, Ba accumulation was found to be the highest in roots followed by leaves and stems. The correlation coefficients (r) of Ba between plants and soils were found to be significantly positively correlated, with the highest correlation being between roots-soils (r=0.922, p
The association of arbuscular mycorrhiza fungi (AMF) and roots undoubtedly
gives positive advantages to the host plant. However, heavily fertilised soil such as in oil
palm plantation, inhibit the growth of mycorrhiza. Thus, the aim of this research is to
distinguish and quantify the availability of AMF population and propagules at different sites
of an oil palm plantation by Most Probable Number (MPN) assay. In addition, root infection
method was employed to observe host compatibility through the propagation of AMF using
two different types of hosts, monocotyledon (Echinochloa cruss-galli) and dicotyledon
(Vigna radiata). Three different locations at an oil palm plantation were chosen for
sampling. Each location was represented by a distinctive soil series, and were further
divided into two sites, that is canopy and midway area. Midway site had a greater
population of AMF compared to canopy. The result showed that different environments
affect the availability of AMF in the soil. Higher number of AMF infection observed in
monocotyledon host suggests that the fibrous root system provide a better association
with mycorrhiza.
Red seaweed Gracilaria, one of the largest genus in Division Rhodophyta inhabits Sarawak coastal water. This study was designed to identify the species of Gracilaria using morphological approach and to assess selected water quality parameters in Gracilaria habitats. Three field samplings were carried out in Santubong and Asajaya, Sarawak from November 2013 to December 2014. Overall, three species were identified namely Gracilaria changii, G. blodgettii and G. coronopifolia, attached to net of cage culture in Santubong and root of mangrove trees in Asajaya. In addition, three different taxa of aquatic macroinvertebrates (polychaete, small crab, bivalve) and single species of red seaweed (Acanthophora sp.) were observed in Gracilaria assemblages. An estimate of 37% to 40% of the upper part of the cage net in Santubong was covered by seaweeds and only 16% to 20% in Asajaya's mangrove. The study had provided better information on identification of Gracilaria and their habitat in Sarawak. Future work involving DNA barcoding of each species is in progress.
Treatment with hypovirulent binucleate Rhizoctonia (HBNR) isolates induced systemic resistance against anthracnose infected by Colletotrichum orbiculare in cucumber, as there were no direct interaction between HBNR and C. orbiculare. This is because of the different distances between HBNR and C. orbiculare, where the root was treated with HBNR isolate and C. orbiculare was challenged and inoculated in leaves or first true leaves were treated with HBNR isolate and C. orbiculare was challenged and inoculated in second true leaves. The use of barley grain inocula and culture filtrates of HBNR significantly reduced the lesion diameter compared to the control (p = 0.05). The total lesion diameter reduction by applying barley grain inoculum of HBNR L2, W1, W7, and Rhv7 was 28%, 44%, 39%, and 40%, respectively. Similar results was also observed in treatment using culture filtrate, and the reduction of total lesion diameter by culture filtrate of HBNR L2, W1, W7, and Rhv7 was 45%, 46%, 42%, and 48%, respectively. When cucumber root was treated with culture filtrates of HBNR, the lignin was enhanced at the pathogen penetration, which is spread along the epidermis tissue of cucumber hypocotyls. Peroxidase activity in hypocotyls in the treated cucumber plant with culture filtrates of HBNR significantly increased before and after inoculation of pathogens as compared to the control. Significant enhancement was also observed in the fast-moving anodic peroxidase isozymes in the treated plants with culture filtrates of HBNR. The results showed the elicitor(s) contained in culture filtrates in HBNR. The lignin deposition as well as the peroxidase activity is an important step to prevent systemically immunised plants from pathogen infection.
Many medicinal plants have been used for centuries in daily life to treat microbial diseases all over the world. In this study, the in vitro antibacterial activity of aqueous and ethanol root extracts of Thespesia populnea Linn were investigated. Antimicrobial properties of T. populnea Linn was evaluated against five pathogenic bacteria and two fungi. Disc diffusion method and minimum inhibitory concentration (MIC) were determined by broth serial dilution method. The ciprofloxacin (5 μg/ml) and flucanozole (100 units/disc) were used as positive controls for bacteria and fungi respectively. Different concentrations (50, 100, 150 μg/ml) of ethanolic and aqueous root extracts of T. populnea were checked for the dose dependent antibacterial activity. Thespesia populnea showed broad spectrum antimicrobial activity against gram positive and gram negative bacteria and maximum inhibition by ethanolic extract was observed at higher dose (250 μg/ml) as 27±0.2mm. The MIC of the ethanol extract was 10 μg/ml for Staphylococcus aureus and 750 μg/ml for Candida albicans. The antifungal activity offered against S. aureus by the ethanolic extract is more than the aqueous extract. The results concluded that the anti-microbial activity of T. populnea was dose dependent. As the concentration increased the inhibition zone also increased. Flavonoids and tannins present in the extracts may be responsible for the antimicrobial activity.
Eurycoma longifolia, locally known as 'Tongkat Ali' is a popular local medicinal plant that possess a lot of medicinal properties as claimed traditionally, especially in the treatment of malaria. The claims have been proven scientifically on isolated compounds from the plant. The present study is to investigate the anti malaria properties of Eurycoma longifolia standardized extract (root) (TA164) alone and in combination with artemisinin in vivo. Combination treatment of the standardized extract (TA164) with artemisinin suppressed P. yoelii infection in the experimental mice. The 4 day suppressive test showed that TA164 suppressed the parasitemia of P. yoelii-infected mice as dose dependent manner (10, 30 and 60 mg/kg BW) by oral and subcutaneous treatment. By oral administration, combination of TA164 at 10, 30 and 60 mg/kg BW each with artemisinin respectively showed a significant increase in the parasitemia suppression to 63, 67 and 80 percent as compared to artemisinin single treatment (31%). Using subcutaneous administration, at 10 mg/kg BW of TA164 in combination with 1.7 mg/kg BW of artemisinin gave a suppression of 80% of infection. This study showed that combination treatment of TA164 with artemisinin gives a promising potential anti malaria candidate using both oral and subcutaneous route, the later being the most potent.
Single-node leafy stem cuttings of Shorea leprosula Miq. were subjected to a high, intermediate or low irradiance treatment for 16 weeks in an enclosed mist propagation system. Before rooting, maximum photosynthesis of the cuttings occurred at an irradiance of 400 micro mol m(-2) s(-1). Although none of the irradiance treatments affected the number of roots produced per cutting, the numbers of cuttings that formed roots were 50 and 30% in the high irradiance (diurnal range of 0-658 micro mol m(-2) s(-1)) and low irradiance (diurnal range of 0-98 micro mol m(-2) s(-1)) treatments, respectively, compared with 62% in the intermediate irradiance treatment (diurnal range of 0-360 micro mol m(-2) s(-1)). Low rooting frequency of cuttings in the high irradiance treatment was associated with water deficits (maximum leaf-to-air vapor pressure deficit (VPD) = 3.6 kPa), whereas cuttings in the low irradiance treatment had a low rooting frequency because they were below the light compensation point most of the time. In the intermediate irradiance treatment, cuttings withstood a daily maximum VPD of 1-2 kPa and recovered overnight from the previous day's deficit, as indicated by higher relative water content (RWC) and stomatal conductance (g(s)) in the morning than in the previous afternoon and evening. Higher RWC and g(s) of cuttings in all treatments on Days 14 and 21 compared with Day 8 probably indicated recovery from water deficit following severance and insertion of the cuttings in rooting medium. There were negative relationships between stem volume of cuttings and both number of cuttings that rooted and number of roots per cutting.
Clarifying the dynamics of fine roots is critical to understanding carbon and nutrient cycling in forest ecosystems. An optical scanner can potentially be used in studying fine-root dynamics in forest ecosystems. The present study examined image analysis procedures suitable for an optical scanner having a large (210 mm × 297 mm) root-viewing window. We proposed a protocol for analyzing whole soil images obtained by an optical scanner that cover depths of 0-210 mm. We tested our protocol using six observers with different experience in studying roots. The observers obtained data from the manual digitization of sequential soil images recorded for a Bornean tropical forest according to the protocol. Additionally, the study examined the potential tradeoff between the soil image size and accuracy of estimates of fine-root dynamics in a simple exercise. The six observers learned the protocol and obtained similar temporal patterns of fine-root growth and biomass with error of 10-20% regardless of their experience. However, there were large errors in decomposition owing to the low visibility of decomposed fine roots. The simple exercise revealed that a smaller root-viewing window (smaller than 60% of the original window) produces patterns of fine-root dynamics that are different from those for the original window size. The study showed the high applicability of our image analysis approach for whole soil images taken by optical scanners in estimating the fine-root dynamics of forest ecosystems.
The root systems of forest trees are composed of different diameters and heterogeneous physiological traits. However, the pattern of root respiration rates from finer and coarser roots across various tropical species remains unknown. To clarify how respiration is related to the morphological traits of roots, we evaluated specific root respiration and its relationships to mean root diameter (D) of various diameter and root tissue density (RTD; root mass per unit root volume; gcm(-3)) and specific root length (SRL; root length per unit root mass; mg(-1)) of the fine roots among and within 14 trees of 13 species from a primary tropical rainforest in the Pasoh Forest Reserve in Peninsular Malaysia. Coarse root (2-269mm) respiration rates increased with decreasing D, resulting in significant relationships between root respiration and diameter across species. A model based on a radial gradient of respiration rates of coarse roots simulated the exponential decrease in respiration with diameter. The respiration rate of fine roots (<2mm) was much higher and more variable than those of larger diameter roots. For fine roots, the mean respiration rates for each species increased with decreasing D. The respiration rates of fine roots declined markedly with increasing RTD and increased with increasing SRL, which explained a significant portion of the variation in the respiration among the 14 trees from 13 species examined. Our results indicate that coarse root respiration in tree species follows a basic relationship with D across species and that most of the variation in fine root respiration among species is explained by D, RTD and SRL. We found that the relationship between root respiration and morphological traits provides a quantitative basis for separating fine roots from coarse roots and that the pattern holds across different species.
The chromosomal aberrations (CA) assay and micronucleus (MN) test were employed to investigate the effect in vitro of zerumbone (ZER) on human chromosomes. ZER is a sesquiterpene compound isolated from the rhizomes of wild ginger, Zingiber zerumbet Smith. The rhizomes of the plant are employed as a traditional medicine for some ailments and as condiments. ZER has been shown to have anti-cancer and apoptosis-inducing properties against various human tumour cells. It has also been shown to be active in vivo against a number of induced malignancies. Studies on ZER genotoxicity in cultured human peripheral blood lymphocytes (PBL) have not been reported so far. Therefore, the present study was undertaken to investigate the ability of ZER to induce chromosomal aberrations and micronuclei formation in human lymphocytes in vitro. Human blood samples were obtained from four healthy, non-smoking males aged 25-35years. Cultures were exposed to the drug for 48h at four final concentrations: 10, 20, 40 and 80 microM. Mitomycin C (MMC) was used as a positive control. The results of chromosomal aberrations assay showed that ZER was not clastogenic, when compared to untreated control, meanwhile MN test results showed a dose-dependent increase in MN formation. The overall clastogenic effect of ZER on human PBL was statistically not significant. In conclusion, ZER is a cytotoxic but not a clastogenic substance in human PBL.
Influences of irrigation frequency on the growth and flowering of chrysanthemum grown under restricted root volume were tested. Chrysanthemum cuttings (Chrysanthemum morifolium "Reagan White") were grown in seedling tray which contained coconut peat in volumes of 73 and 140 cm(3). Plants were irrigated with drip irrigation at irrigation frequencies of 4 (266 mL), 6 (400 mL), and 8 (533 mL) times/day to observe their growth and flowering performances. There was interaction between irrigation frequency and substrate volume on plant height of chrysanthemum. Plants grown in 140 cm(3) substrates and irrigated 6 times/day produced the tallest plant of 109.25 cm. Plants irrigated 6 and 8 times/day had significantly higher level of phosphorus content in their leaves than those plants irrigated 4 times/day. The total leaf area, number of internodes, leaf length, and leaf width of chrysanthemums grown in 140 cm(3) substrate were significantly higher than those grown in 73 cm(3) substrate. The numbers of flowers were affected by both irrigation frequencies and substrate volumes. Chrysanthemums irrigated 8 times/day had an average of 19.56 flowers while those irrigated 4 times/day had an average of 16.63 flowers. Increasing irrigation frequency can improve the growth and flowering of chrysanthemums in small substrate volumes.
To explore the potential of in vitro rapid regeneration, three varieties (Golpaygan-181, Orumieh-1763, and Gorgan-1601) of sainfoin (Onobrychis viciifolia Scop. syn. Onobrychis sativa L.) were evaluated. For the first time, an encapsulation protocol was established from somatic embryogenic callus in torpedo and cotyledonary stages to create artificial seeds. Callus derived from different concentrations of Kinetin (0-2.0 mg L(-1)) and Indole-3-acetic acid (0-2.0 mg L(-1)) was coated with sodium alginate and subsequently cultured either in Murashige and Skoog (MS) medium or in soil substrate. Adventitious shoots from synthetic beads developed into rooting in full and half strength MS medium supplemented with various concentrations of auxin and cytokinin. Prolonged water conservation of black and red soils (1:1) had the highest rate of survival plantlets in the acclimatization process. Diverse resistance techniques in Onobrychis viciifolia were evaluated when the plants were subjected to water deficiency. Higher frequency of epicuticular waxes was observed in in vivo leaves compared to in vitro leaves. Jagged trichomes nonsecreting glands covered by spines were only observed in the lower leaf side. Ultimately, stomata indices were 0.127 (abaxial), 0.188 (adaxial) in in vivo and 0.121 (abaxial), 0.201 (adaxial) in in vitro leaves.