Testa is involved in the control of storage mobilisation in seeds of <Emphasis Type="Italic">Sesbania virgata</Emphasis> (Cav.) Pers., a tropical legume tree from of the Atlantic Forest

Seeds of Sesbania virgata (Cav.) Pers. (Leguminosae) contain galactomannan as a cell wall storage polysaccharide in the endosperm. After germination, it is hydrolysed by three enzymes: α-galactosidase (EC 3.2.1.22), endo-β-mannanase (EC 3.2.1.78) and β-mannosidase (EC 3.2.1.25). This work aimed at studying the role of the testa (seed coat) on galactomannan degradation during and after germination. Seeds were imbibed in water, with and without the testa, and used to evaluate the effect of this tissue on storage mobilisation, as well as its possible role in the galactomannan hydrolases activities. Immunocytochemistry and immunodotblots were used to follow biochemical events by detecting and localising endo-β-mannanase in different tissues of the seed. Endo-β-mannanase and α-galactosidase activities were found in the testa and latter in the endosperm during galactomannan degradation. The former enzyme was immunologically detected in the testa, mainly during germination. The absence of the testa during imbibition led to the anticipation of protein mobilisation and increased of the α-galactosidase activity and galactomannan degradation. Thus, the testa appears to play a role during storage mobilisation in the legume seed of S. virgata probably by participating in the control of the production, modification and/or storage of the hydrolases.     

Effect of brown algae metabolites on the synthesis of O-glycosyl hydrolases by bacteria degrading the thallus of <Emphasis Type="Italic">Fucus evanescens</Emphasis>

The effect of fucoidan, extractive substances from Fucus evanescens and the Laminaria cichorioides protein (an inhibitor of endo-1→3-β-D-glucanase) on the degradation of F. evanescens thallus and on the growth of bacteria involved in this process was studied. The complex of O-glycosyl hydrolases and the level of their enzymatic activity in bacteria cultivated under various conditions depended significantly on the composition of the growth medium. The highest taxonomic diversity was observed for bacteria isolated from the thallus degraded in the control medium (sea water). These bacteria were characterized by very low levels of activity of the enzymes degrading polysaccharides (fucoidan, laminaran, and pustulan). In the presence of 1→3-β-D-glucanase inhibitors, the taxonomic diversity of the microorganisms degrading F. evanescens thallus was decreased, and the activity of O-glycosyl hydrolases (particularly, of fucoidan hydrolases) was increased.     

Ecophysiological Variabilities in Ectohydrolytic Enzyme Activities of Some <Emphasis Type="Italic">Pseudoalteromonas</Emphasis> Species, <Emphasis Type="Italic">P. citrea,P. issachenkonii</Emphasis>, and <Emphasis Type="Italic">P. nigrifaciens</Emphasis>

The ecophysiological variabilities in the ectohydrolytic enzyme profiles of the three species of Pseudoalteromonas, P. citrea, P. issachenkonii, and P. nigrifaciens, have been investigated. Forty-one bacteria isolated from several invertebrates, macroalgae, sea grass, and the surrounding water exhibited different patterns of hydrolytic enzyme activities measured as the hydrolysis of either native biopolymers or fluorogenic substrates. The activities of the following enzymes were assayed: proteinase, tyrosinase, lipase, amylase, chitinase, agarase, fucoidan hydrolase, laminaranase, alginase, pustulanase, cellulase, β-glucosidase, α- and β-galactosidases, β-N-acetylglucosaminidase, β-glucosaminidase, β-xylosidase, and α-mannosidase. The occurrence and cell-specific activities of all enzymes varied over a broad range (from 0 to 44 μmol EU per hour) and depended not only on taxonomic affiliation of the strain, but also on the source/place of its isolation. This suggests ‘specialization’ of different species for different types of polymeric substrates as, for example, all strains of P. citrea and P. issachenkonii hydrolyzed alginate and laminaran, while strains of P. nigrifaciens were lacking the ability to hydrolyze most of the algal polysaccharides. The incidence of certain enzymes such as fucoidan hydrolases, alginate lyases, agarases, and α-galactosidases might be strain specific and reflect its particular ecological habitat. Received: 15 February 2002 / Accepted: 27 March 2002     

Isolation and properties of extracellular β-xylosidases from fungi <Emphasis Type="Italic">Aspergillus japonicus</Emphasis> and <Emphasis Type="Italic">Trichoderma reesei</Emphasis>

Homogeneous β-xylosidases with molecular mass values 120 and 80 kDa (as shown by SDS-PAGE), belonging to the third family of glycosyl hydrolases, were isolated by anion-exchange, hydrophobic, and gel-penetrating chromatography from enzyme preparations based on the fungi Aspergillus japonicus and Trichoderma reesei, respectively. The enzymes exhibit maximal activity in acidic media (pH 3.5–4.0), and temperature activity optimum was 70°C for the β-xylosidase of A. japonicus and 60°C for the β-xylosidase of T. reesei. Kinetic parameters of p-nitrophenyl β-xylopyranoside and xylooligosaccharide hydrolysis by the purified enzymes were determined, which showed that β-xylosidase of A. japonicus was more specific towards low molecular weight substrates, while β-xylosidase of T. reesei preferred high molecular weight substrates. The competitive type of inhibition by reaction product (xylose) was found for both enzymes. The interaction of the enzymes of different specificity upon hydrolysis of glucurono- and arabinoxylans was found. The β-xylosidases exhibit synergism with endoxylanase upon hydrolysis of glucuronoxylan as well as with α-L-arabinofuranosidase and endoxylanase upon hydrolysis of arabinoxylan. Addition of β-xylosidases increased efficiency of hydrolysis of plant raw materials with high hemicellulose content (maize cobs) by the enzymic preparation Celloviridine G20x depleted of its own β-xylosidase.     

Microbial hydrolytic enzyme activities in deep-sea sediments

The potential hydrolysis rates of five different hydrolytic enzymes were determined in deep-sea sediments from the northeast Atlantic (BIOTRANS area) in March 1992. Fluorogenic substrates were used to assay extracellular α- and β-glucosidase, chitobiase, lipase and aminopeptidase. The potential activity of most of the enzymes investigated decreased to a minimum within the upper two centimetre range, whereas aminopeptidase was high over the upper five centimetre range. Exceptions were found when macrofaunal burrows occurred in the cores, always increasing the activities of some hydrolases, and therefore indicating the impact of bioturbation on degradation rates. The most striking feature of the investigated enzyme spectrum was the 50–2000 times higher specific activity of the aminopeptidase, compared with the other hydrolases. The activity of hydrolytic enzymes most likely reflects the availability of their respective substrates and is not a function of bacterial biomass.     

Compositional and enzymatic changes in guava (Psidium guajava L.) fruits during ripening

Changes in chemical composition and hydrolytic enzyme activities in guava fruits cv. Lucknow-49 have been reported at four different stages of maturity, viz., mature green (MG), color turning (CT), ripe (R) and over ripe (OR). Chlorophyll content decreased, while carotenoid content increased with advancement of ripening. Starch content decreased with concomitant increase in alcohol soluble sugars. The cell wall constituents viz., cellulose, hemicellulose, and lignin decreased up to R stage, while the pectin content decreased throughout up to OR stage. Among the cell wall hydrolyzing enzymes, polygalacturonase (PG) and cellulase exhibited progressive increase in activity throughout ripening, while pectin methyl esterase (PME) activity increased up to CT stage and then decreased up to OR stage. The maximum increase in the activities of cell wall hydrolysing enzymes was observed between MG and CT stages. The activities of starch hydrolyzing enzymes, α-amylase and β-amylase decreased significantly with advancement of ripening. These changes in the activities of hydrolyzing enzymes could be considered good indicators of ripening in guava.     

A laboratory study on digestive processes in the Antarctic krill,Euphausia superba,with special regard to chitinolytic enzymes

Feeding experiments of 9, 14 and 20 days duration were carried out on the Antarctic krill, Euphausia superba. Two groups were fed with the chitinous diatom Cyclotella cryptica and the non-chitinous green algae Dunaliella bioculata, respectively. A control group remained unfed. The time courses of the activities of endo- and exochitinase in the stomach and the midgut gland were compared with those of the digestive enzymes protease, cellulase (1,4-β-d-glucanase) and laminarinase (1,3-β-d-glucanase). Specific activities of all enzymes were higher in the stomach than in the midgut gland. Characteristic time courses of activity were evident after 4 days. In starved animals, enzyme activities decreased to a minimum after 4 days and recovered within 14 days to initial values. In the stomach, the activities of endo- and exochitinase increased when krill were fed on Cyclotella. For animals fed with Dunaliella, activities stayed constant or decreased slightly. The results confirm chitinases as digestive enzymes and, therefore, the capability of krill to utilize various food sources. Accepted: 4 October 1998     

Activities of sulfhydryl-related and phenylpropanoid-synthesizing enzymes during the germination ofArabidopsis thaliana seeds

Seeds ofArabidopsis thaliana were sowed in a Murashige and Skoog’s medium and incubated in a tissue culture chamber at 26.2°C. They appeared to germinate at 60–72 hours after sowing. We measured the time-course activities of sulfhydryl-related enzymes such as thioredoxin, thioltransferase (glutaredoxin), thioredoxin reductase, and glutathione reductase. The activities of two enzymes, phenylalanine ammonia-lyase and tyrosine ammonia-lyase, which are involved in phenylpropanoid synthesis, were also measured and compared. Phenylalanine ammonia-lyase activity increased from 48 hours after sowing, whereas tyrosine ammonia-lyase activity increased transiently at the early stage and decreased slightly afterwards. Thioredoxin activity gradually decreased during the germination process. However, thioltransferase activity increased drastically from 60 hours after sowing. Thioredoxin reductase and glutathione reducatase increased rapidly from 36 hours after sowing. Thioredoxin activity was found to be relatively high in the dormant seeds.     

Selenium-mediated differential response of β-glucosidase and β-galactosidase of germinatingTrigonella foenum-graecum

β-Glucosidase and β-galactosidase activity profile tested in different seeds during 24 h germination revealed reasonably high levels of activity inVigna radiata, Cicer arietinum, andTrigonella foenum-graecum. In all seeds tested, β-galactosidase activity was, in general, higher than that of β-glucosidase.T. foenum-graecum seedlings exhibited maximal total and specific activities for both the enzymes during 72 h germination. Se supplementation as Na₂SeO₃ up to 0.75 ppm was found to be beneficial to growth and revealed selective enhancement of β-galactosidase activity by 40% at 0.5 ppm Se. The activities of both the enzymes drastically decreased at 1.0 ppm level of Se supplementation. On the contrary, addition of Na₂SeO₃ in vitro up to 1 ppm to the enzyme extracts did not influence these activities. Hydrolytic rates of β-glucosidase in both control and Se-supplemented groups were enhanced by 20% with 0.05M glycerol in the medium and 30% at 0.1M glycerol. The rates were marginally higher in Se-supplemented seedlings than the controls, irrespective of added glycerol in the medium. In contrast, hydrolysis by β-galactosidase showed a trend of decrease in Se-supplemented seedlings compared to the control, when glycerol was present in the medium. Addition of Se in vitro in the assay medium showed no difference in the hydrolytic rate by β-galactosidase when compared to control, while the activity of β-glucosidase declined by 50%. Se-grown seedlings showed an enhancement of transglucosidation rate by 40% in the presence of 0.1M glycerol. The study reveals a differential response to Se among the β-galactosidase and β-glucosidase ofT. foenumgraecum with increase in the levels of β-galactosidase activity.     

Soil enzyme activities and organic matter composition in a turfgrass chronosequence

Highly managed turfgrass systems accumulate considerable soil organic C, which supports a diverse and robust soil microbial community. Degradation of this soil organic C is mediated by a suite of soil enzymes. The relationship between these enzyme activities and the quality of soil organic C is central to understanding the dynamics of soil organic matter. We examined the activities of several soil enzymes involved in microbial C acquisition, including β-glucosidase, N-acetyl-β-glucosaminidase, cellulase, chitinase, and phenol oxidase, and characterized the chemical composition of soil organic matter using Fourier transform infrared spectroscopy (FTIR) in a turfgrass chronosequence (1–95 years old) and adjacent native pines. Non-metric multidimensional scaling analysis showed that the chemical composition of soil organic matter varied with turf age and land use (turf versus pines). Using the polysaccharide peak (1,060 cm⁻¹) as a reference, both aliphatic (2,930 cm⁻¹) and carboxylic (1,650 and 1,380 cm⁻¹) compounds increased with turf age, indicating that soil organic matter became more recalcitrant. Soil enzyme activities per unit soil mass increased with turf age and were correlated to soil C content. Most soil enzyme activities in native pines were similar to those in young turf, but the cellulase activity was similar to or greater than the activity in old turfgrass systems. On a soil C basis, however, the activities of N-acetyl-β-glucosaminidase and cellulase decreased with turf age; this reduction was correlated to the relative changes in the chemical composition of soil organic matter. We observed that the chemical composition of soil organic matter was significantly correlated with the enzyme activity profile when expressed per unit microbial biomass C, but not per unit soil organic C. Our results suggest that chemical composition of soil organic matter changes with turf age and this change partially determines the relative abundance of C-degrading soil enzymes, likely through the influence on microbial community composition.     

Serum steroid hormone levels in relation to the reproductive cycle ofSebastes taczanowskii andS. schlegeli

Synopsis Changes in serum steroid hormones were studied during the reproductive cycle of two viviparous rockfishes of the genusSebastes. During the annual reproductive cycle of femaleS. taczanowskii, serum levels of estradiol-17β (E₂) gradually increased from their lowest in August to maximal levels towards the end of vitellogenesis in February, and rapidly returned to low levels in pregnant females in April and May. Serum progesterone (prog) fluctuated at low levels throughout the annual reproductive cycle, while 17α, 20β-dihydroxy-4-pregnen-3-one (17α, 20β-diOHprog) levels were high during gestation in May. Serum levels of these steroids were also measured in femaleS. schlegeli at weekly intervals during late vitellogenesis, gestation and post-parturition. In pregnant females, E₂ was high during vitellogenesis, then decreased and remained low throughout gestation. The 17α, 20β-diOHprog levels were low during vitellogenesis, then rapidly increased and remained high during gestation. On the other hand, prog values remained low, with temporal peaks coinciding with the peak of 17α, 20β-diOHprog during gestation. Based on these results and the literature, it is suggested that E₂ is the most important steroid involved in vitellogenesis and that 17α, 20β-diOHprog may play an important role in final oocyte maturation and subsequent gestation.     

Cloning and identification of novel cellulase genes from uncultured microorganisms in rabbit cecum and characterization of the expressed cellulases

A metagenomic cosmid library was prepared in Escherichia coli from DNA extracted from the contents of rabbit cecum and screened for cellulase activities. Eleven independent clones expressing cellulase activities (four endo-β-1,4-glucanases and seven β-glucosidases) were isolated. Subcloning and sequencing analysis of these clones identified 11 cellulase genes; the encoded products of which shared less than 50% identities and 70% similarities to cellulases in the databases. All four endo-β-1,4-glucanases and all seven β-glucosidases, respectively, belonged to glycosyl hydrolase family 5 (GHF 5) and family 3 (GHF 3) and formed two separate branches in the phylogenetic tree. Ten of the 11 cloned cellulases exhibited highest activities at pH 5.5 ∼ 7.0 and 40 ∼ 55°C, a condition similar to that in the rabbit cecum. All the four endo-β-1,4-glucanases could hydrolyze a wide range of β-1,4-, β-1,4/β-1,3- or β-1,3/β-1,6-linked polysaccharides. One endo-β-1, 4-glucanase gene, umcel5G, was overexpressed in E. coli, and the purified recombinant enzyme was characterized in detail. The enzymes cloned in this work represented at least some of the cellulases operating efficiently in the rabbit cecum. This work provides the first snapshot on the cellulases produced by bacteria in rabbit cecum.     

Protective effect of betaine on changes in the levels of lysosomal enzyme activities in heart tissue in isoprenaline-induced myocardial infarction in Wistar rats

Myocardial infarction is one of the most common manifestations of cardiovascular disease. In the present study, we investigated the protective effect of betaine, a potent lipotropic molecule, on changes in the levels of lysosomal enzymes and lipid peroxidation in isoprenaline-induced myocardial infarction in Wistar rats, an animal model of myocardial infarction in man. Male albino Wistar rats were pretreated with betaine (250 mg/kg body weight) daily for a period of 30 days. After the treatment period, isoprenaline (11 mg/100 g body weight) was intraperitoneally administered to rats at intervals of 24 h for 2 days. The activities of lysosomal enzymes (β-glucuronidase, β-galactosidase, β-glucosidase, and acid phosphatase) were significantly (p < 0.05) increased in plasma with a concomitant decline in the activities of these enzymes in heart tissue of isoprenaline-administered rats. Also, the level of lipid peroxidation was higher in heart lysosomes of isoprenaline-injected rats. Pretreatment with betaine daily for a period of 30 days to isoprenaline-induced rats prevented the changes in the activities of these lysosomal enzymes. Oral treatment with betaine (250 mg/kg body weight) to normal control rats did not show any significant effect in all the biochemical parameters studied. Thus, the results of our study show that betaine protects the lysosomal membrane against isoprenaline-induced myocardial infarction. The observed effects might be due to the free radical-scavenging and membrane-stabilizing properties of betaine.     

Alterations in the photosynthetic pigments and antioxidant machineries of red pepper (<Emphasis Type="Italic">Capsicum annuum</Emphasis> L.) seedlings from gamma-irradiated seeds

To characterize the stimulatory effects of low-dose gamma radiation on early plant growth, we investigated alterations in the photosynthesis and antioxidant capacity of red pepper (Capsicum annuum L.) seedlings produced from gamma-irradiated seeds. For two cultivars (Yeomyung and Joheung), three irradiation groups (2, 4, and 8 Gy, but not 16 Gy) showed enhanced development, although Fv/Fm, the maximum photochemical efficiency of Photosystem II (PSII), did not differ significantly among any of the four groups. In contrast, values for 1/Fo — 1/Fm, i.e., a measure of functional PSII content, decreased in the irradiated groups of ‘Yeomyung’ but increased in those of ‘Joheung’. Pigment analyses and enzyme activity assays revealed that irradiation altered the compositions of photosynthetic pigments (chlorophylls and carotenoids) as well as the activities of antioxidant enzymes (superoxide dismutase and glutathione reductase). However, these shifts were not directly related to the increase in early growth, although they were cultivar-and developmental stage-dependent In addition, the effects of irradiation on the enzymatic activities measured here were at opposition between the two cultivars.     

Effect of abscisic acid on galactomannan degradation and endo-β-mannanase activity in seeds of Sesbania virgata (Cav.) Pers. (Leguminosae)

Seeds of Sesbania virgata (Cav.) Pers. (Leguminosae) have an endosperm which accumulates galactomannan as a storage polysaccharide in the cell walls. After germination, it is hydrolysed by three enzymes: α-galactosidase (EC 3.2.1.22), endo-β-mannanase (EC 3.2.1.78) and β-mannosidase (EC 3.2.1.25). This work aimed at studying the effect of abscisic acid (ABA) on galactomannan degradation during and after germination. Seeds were imbibed in water or in 10⁻⁴ M ABA, and used to evaluate the effect of exogenous and endogenous ABA. Tissue printing was used to follow biochemical events by detecting and localising endo-β-mannanase in different tissues of the seed. The presence of exogenous ABA provoked a delay in the cellular disassembly of the endosperm and disappearance of endo-β-mannanase in the tissue. This led to a delay in galactomannan degradation. The testa (seed coat) of S. virgata contains endogenous ABA, which decreases ca. fourfold during storage mobilisation after germination, permitting the galactomannan degradation in the endosperm. Furthermore, endo-β-mannanase was immunolocalised in the testa, which has a living cell layer. The ABA appears to modulate storage mobilisation in the legume seed of S. virgata, and a cause–effect relationship between ABA (probably through testa) and activities of hydrolases is proposed.     

A fluorimetric method for measuring the activity of soil enzymes

Summary A fluorimetric method is described for the measurement of the activity of a range of soil enzymes. The method is based on the measurement of 4-methylumbelliferone (MUB), a fluorescent product liberated on hydrolysis of the enzyme substrate. The main advantage of the method over colorimetric techniques is that separation of MUB from the soil is unnecessary and the method is therefore suitable for routine, automated analyses. The method was used to measure the activity of β-cellobiase, β-galactosaminidase, β-glucosidase and β-xylosidase over a wide range of substrate concentration and in a range of soils. Kinetic parameters are reported for these enzymes. The method was also shown to be suitable for the assay of arylsulphatase and acid and alkaline phosphatase in soil. The technique should be applicable to a wide range of soil hydrolases, using the same assay methods.     

Metabolic changes in tomato fruits and seeds after viral, bacterial and fungal infections

The metabolic changes in tomato fruits and seeds separately infected with cucumber mosaic virus, Pseudomonas syringae pv. tomato or Botrytis cinerea were investigated cytochemically. The changes of peroxidase (E.C. 1.11.1.7) and β-glucosidase (E.C. 3.2.1.21) were investigated biochemically as well. Tomato fruits were involved in the study because of their high economic value. Tomato seeds were investigated since they have been used most extensively as a model system for studying the physiology and biochemistry of seed development. The diseases caused by the pathogens under study are of special importance for yield reduction in tomato. The three pathogens provoked local changes in the activities of enzymes under study that affect the infected pericarp tissues and neighboring seeds. It was established non-specific and specific responses. The non-specific responses of invaded tissues were expressed as a local enhancement of peroxidase activity in both pericarp tissues and seeds as well as a decrease in activities of: i. enzymes taking part in aerobic and anaerobic respiration, ii. hydrolases esterase and acid phosphatase involved in the basic metabolism as well as an enhancement of their activities in neighboring tissues. Furthermore, it was observed an enhancement of α-galactosidase activity in infected area was observed. The specific responses depending on the type of the pathogen consisted in an enhancement of glucose-6-phosphate dehydrogenase activity by virus infection and an increase of β-glucosidase activity by fungal invasion.     

Xylanases of anaerobic fungus <Emphasis Type="Italic">Anaeromyces mucronatus</Emphasis>

The anaerobic fungus Anaeromyces mucronatus KF8 grown in batch culture on M10 medium with rumen fluid and microcrystalline cellulose as carbon source produced a broad range of enzymes requisite for degradation of plant structural and storage saccharides including cellulase, endoglucanase, xylanase, α-xylosidase, β-xylosidase, α-glucosidase, β-glucosidase, β-galactosidase, mannosidase, cellobiohydrolase, amylase, laminarinase, pectinase and pectate lyase. These enzymes were detected in both the intra- and extracellular fractions, but production into the medium was prevalent with the exception of intracellular β-xylosidase, chitinases, N-acetylglucosaminidase, and lipase. Xylanase activity was predominant among the polysaccharide hydrolases. Extracellular production of xylanase was stimulated by the presence of cellobiose and oat spelt xylan. Zymogram of xylanases of strain KF8 grown on different carbon sources revealed several isoforms of xylanases with approximate molar masses ranging from 26 to 130 kDa.     

Correlation between influence of polysaccharides on hydrolase activity and their antiviral effect in tobacco leaves

The activities of hydrolases (acid phosphatase, RNase, and proteases) in healthy and tobacco mosaic virus-infected leaves of Nicotiana tabacum L. var. Samsun, both untreated and treated with polysaccharides (PS) (1,3;1,6-β-D-glucan, fucoidan, and κ/β-carrageenan), were determined. The PS lead to substantial increase in the hydrolase level. The percentage of viral particles undergoing destructive change also increases in leaves treated with PS 24 h before infection. We suppose that the PS-mediated hydrolase activation promotes intracellular destruction of the viral particles and, thus, comprises one of the PS-induced protective mechanisms limiting intracellular viral accumulation.     

Comparison of the effects of fungal endophyte Gilmaniella sp. and its elicitor on Atractylodes lancea plantlets

The effects of the endophytic fungus Gilmaniella sp. and its elicitor on the defense and metabolic responses of host plants Atractylodes lancea were investigated, in order to understand how to utilize endophytic fungi and their elicitor resources better. The results showed that the promotion effect of the fungus on the growth of host plantlets was much better than that of its elicitor. Both fungus and elicitor enhanced defense-related enzyme activities. In fungus-inoculated groups, phenylalanine ammonia lyase and polyphenol oxidase activities increased slowly, and reached a maximum level during the later stages, whereas peroxidase activity peaked in the first few days. Additionally, the activities of chitinase and β-1,3-glucanase were significantly higher than those of the control plants. In elicitor-treated groups, however, most of the enzymes were activated during the early stage, and their highest levels were generally lower than those of the fungus-inoculated groups. Compared with the elicitor, fungal infection improved the photosynthetic rate of the host, and increased carbohydrate levels as well as chlorophyll content in host leaves. The total content of the four main components of volatile oil was also increased in elicitor-treated groups, but there was no particular pattern in this increase. Meanwhile, in the fungus-inoculated groups, the content of atractylone significantly increased with time, while the content of β-eudesmol decreased. These results indicated that fungal elicitor could substantially improve the total content of volatile oil, while the fungus could more effectively enhance the quality of herbal medicines.