Glucosinolate-containing Plant Research Articles

Suppression of Sclerotium cepivorum Using Biofumigation By Glucosinolate-Containing Plant and Cyanogenic Bacteria اخماد فطر الاسکيرولشيم سيبوفورم باستخدام التبخير الحيوي بالنباتات المحتوية على الجلوکوسينولات والبکتيريا المنتجة لسيانيد الهيدروجين

In dual culture antagonism test, six of eight cyanogenic bacteria were able to reduce the growth of Sclerotium cepivorum. The strains Glocoacetobacter diazotrophicus and Paenibacillus. polymyxa GQ375783.1were the most effective bacteria that recorded the highest inhibition of S. cepivorum mycelial growth by 75% and 70% respectively. Based on HCN production, two strains recorded complete inhibition of fungal growth. Moreover, the ground tissues of the nasturtium plant were able to inhibit the mycelium growth of S. cepivorum by 100%. Results of the greenhouse experiment emphasized that white-rot incidence percentage recorded the lowest values in onion cultivated in soil amended with nasturtium plant and inoculated with G. diazotrophicus plus P. polymyxa GQ375783.1. This treatment recorded the highest values of onion vegetative growth and their yield. Under field conditions, the soil treated with investigated two methods of biofumigation led to suppress white rot disease by the rate of 100% while using each one individually, it reduced disease by the rate of 94.4 and 94.9 % respectively. All onion treatments that inoculated with cyanogenic bacteria showed higher records of DH and N2-ase activity than uninoculated ones. The highest values DH and N2-ase were recorded with soil amended with nasturtium and inoculated with cyanogenic bacterial strains. Additionally, the same treatment gave the highest values of peroxidase and polyphenoloxidase activity, onion NPK content, and the plant's vegetative growth as well as their yield. Therefore, the biofumigation using nasturtium and investigated bacterial strains can be recommended in controlling White-rot disease in onions and increasing their productivity.

A Straightforward Method for Glucosinolate Extraction and Analysis with High-pressure Liquid Chromatography (HPLC)

Glucosinolates are a well-studied and highly diverse class of natural plant compounds. They play important roles in plant resistance, rapeseed oil quality, food flavoring, and human health. The biological activity of glucosinolates is released upon tissue damage, when they are mixed with the enzyme myrosinase. This results in the formation of pungent and toxic breakdown products, such as isothiocyanates and nitriles. Currently, more than 130 structurally different glucosinolates have been identified. The chemical structure of the glucosinolate is an important determinant of the product that is formed, which in turn determines its biological activity. The latter may range from detrimental (e.g., progoitrin) to beneficial (e.g., glucoraphanin). Each glucosinolate-containing plant species has its own specific glucosinolate profile. For this reason, it is important to correctly identify and reliably quantify the different glucosinolates present in brassicaceous leaf, seed, and root crops or, for ecological studies, in their wild relatives. Here, we present a well-validated, targeted, and robust method to analyze glucosinolate profiles in a wide range of plant species and plant organs. Intact glucosinolates are extracted from ground plant materials with a methanol-water mixture at high temperatures to disable myrosinase activity. Thereafter, the resulting extract is brought onto an ion-exchange column for purification. After sulfatase treatment, the desulfoglucosinolates are eluted with water and the eluate is freeze-dried. The residue is taken up in an exact volume of water, which is analyzed by high-pressure liquid chromatography (HPLC) with a photodiode array (PDA) or ultraviolet (UV) detector. Detection and quantification are achieved by conducting comparisons of the retention times and UV spectra of commercial reference standards. The concentrations are calculated based on a sinigrin reference curve and well-established response factors. The advantages and disadvantages of this straightforward method, when compared to faster and more technologically advanced methods, are discussed here.

A Straightforward Method for Glucosinolate Extraction and Analysis with High-pressure Liquid Chromatography (HPLC).

Glucosinolates are a well-studied and highly diverse class of natural plant compounds. They play important roles in plant resistance, rapeseed oil quality, food flavoring, and human health. The biological activity of glucosinolates is released upon tissue damage, when they are mixed with the enzyme myrosinase. This results in the formation of pungent and toxic breakdown products, such as isothiocyanates and nitriles. Currently, more than 130 structurally different glucosinolates have been identified. The chemical structure of the glucosinolate is an important determinant of the product that is formed, which in turn determines its biological activity. The latter may range from detrimental (e.g., progoitrin) to beneficial (e.g., glucoraphanin). Each glucosinolate-containing plant species has its own specific glucosinolate profile. For this reason, it is important to correctly identify and reliably quantify the different glucosinolates present in brassicaceous leaf, seed, and root crops or, for ecological studies, in their wild relatives. Here, we present a well-validated, targeted, and robust method to analyze glucosinolate profiles in a wide range of plant species and plant organs. Intact glucosinolates are extracted from ground plant materials with a methanol-water mixture at high temperatures to disable myrosinase activity. Thereafter, the resulting extract is brought onto an ion-exchange column for purification. After sulfatase treatment, the desulfoglucosinolates are eluted with water and the eluate is freeze-dried. The residue is taken up in an exact volume of water, which is analyzed by high-pressure liquid chromatography (HPLC) with a photodiode array (PDA) or ultraviolet (UV) detector. Detection and quantification are achieved by conducting comparisons of the retention times and UV spectra of commercial reference standards. The concentrations are calculated based on a sinigrin reference curve and well-established response factors. The advantages and disadvantages of this straightforward method, when compared to faster and more technologically advanced methods, are discussed here.

A thiocyanate-forming protein generates multiple products upon allylglucosinolate breakdown in Thlaspi arvense

Glucosinolates, amino acid-derived thioglycosides found in plants of the Brassicales order, are one of the best studied classes of plant secondary metabolites. Together with myrosinases and supplementary proteins known as specifier proteins, they form the glucosinolate–myrosinase system that upon tissue damage gives rise to a number of biologically active glucosinolate breakdown products such as isothiocyanates, epithionitriles and organic thiocyanates involved in plant defense. While isothiocyanates are products of the spontaneous rearrangement of the glucosinolate aglycones released by myrosinase, the formation of epithionitriles and organic thiocyanates depends on both myrosinases and specifier proteins. Hydrolysis product profiles of many glucosinolate-containing plant species indicate the presence of specifier proteins, but only few have been identified and characterized biochemically. Here, we report on cDNA cloning, heterologous expression and characterization of TaTFP, a thiocyanate-forming protein (TFP) from Thlaspi arvense L. (Brassicaceae), that is expressed in all plant organs and can be purified in active form after heterologous expression in Escherichia coli. As a special feature, this protein promotes the formation of allylthiocyanate as well as the corresponding epithionitrile upon myrosinase-catalyzed hydrolysis of allylglucosinolate, the major glucosinolate of T. arvense. All other glucosinolates tested are converted to their simple nitriles when hydrolyzed in the presence of TaTFP. Despite its ability to promote allylthiocyanate formation, TaTFP has a higher amino acid sequence similarity to known epithiospecifier proteins (ESPs) than to Lepidium sativum TFP. However, unlike Arabidopsis thaliana ESP, its activity in vitro is not strictly dependent on Fe 2+ addition to the assay mixtures. The availability of TaTFP in purified form enables future studies to be aimed at elucidating the structural bases of specifier protein specificities and mechanisms. Furthermore, identification of TaTFP shows that product specificities of specifier proteins can not be predicted based on amino acid sequence similarity and raises interesting questions about specifier protein evolution.

Glucosinolates and biofumigation: fate of glucosinolates and their hydrolysis products in soil

The bioactive hydrolysis products of glucosinolates, particularly the isothiocyanates, can be used to control soil pests and weeds by incorporating glucosinolate-containing plant material in soil—a practice known as biofumigation. The fate of glucosinolates and their hydrolysis products in soil determines both the efficacy and environmental impact of biofumigation. Knowledge of the processes by which these compounds are sorbed, degraded or otherwise lost from the soil is fundamental to developing effective, but environmentally benign biofumigation strategies. Effective biofumigation relies on maximum hydrolysis of the glucosinolate in the plant tissue to generate high isothiocyanate concentrations in the soil after incorporation. This is favoured by maximum cell disruption, by addition of water, and a high soil temperature. Residual glucosinolates are very weakly sorbed, readily leached and are microbially degraded and mineralised in soil. In contrast, isothiocyanates are strongly sorbed by the organic matter in soil, react strongly with nucleophilic groups present in soil, and are prone to volatilization losses in addition to microbial degradation and mineralisation. These loss processes are influenced by soil type, water content and temperature. Using appropriate incorporation strategies, sufficiently high isothiocyanate concentrations (>100 nmol g−1) can be achieved in soil using biofumigation for effective suppression of susceptible pests. The relatively rapid sorption and degradation of the isothiocyanates in the period of days after incorporation minimizes the risks of persistence in the environment or leaching. Biofumigation is therefore a promising technique which can be further developed to form part of IPM (Integrated Pest Management) strategies to reduce reliance on synthetic pesticides with minimal unintended impacts on the environment.

Herbicidal activity of glucosinolate-containing seedmeals

Defatted seedmeals from 15 glucosinolate-containing plant species were analyzed for herbicidal activity by determining inhibition of seedling emergence when added to a sandy loam soil containing wheat and sicklepod seeds at concentrations of 0.1, 0.5, and 1% (w/w). In general, the seedmeals were more phytotoxic to wheat than sicklepod. For wheat, all of the seedmeals significantly inhibited seedling emergence at the 1.0% concentration. At the 0.1% concentration three of the seedmeals (Indian mustard, money plant, and field pennycress) completely inhibited wheat emergence. For sicklepod emergence, eight of the seedmeals were completely inhibitory at the 1% level (Indian mustard, field pennycress, garden rocket, Siberian wallflower, English wallflower, garden cress, sweet alyssum, and evening stock) and four were completely inhibitory at the 0.5% level (brown mustard, garden rocket, English wallflower, and sweet alyssum). Intact glucosinolates and their corresponding hydrolysis products varied among the seedmeals with the highest activity. Major hydrolysis products produced by the seedmeals with the most phytotoxicity, respectively, included 2-propenyl (allyl) isothiocyanate (AITC) by brown mustard seedmeal, allyl thiocyanate and AITC by field pennycress seedmeal, erucin (4-methylthiobutyl isothiocyanate) by arugula seedmeal, 3-butenyl isothiocyanate and lesquerellin (6-methylthiohexyl isothiocyanate) by sweet alyssum seedmeal, and isopropyl isothiocyanate by money plant seedmeal. From our data it appears that both the type and concentration of glucosinolates and their hydrolysis products present in the seedmeals affect seed-emergence inhibition.

Comparative biochemical characterization of nitrile‐forming proteins from plants and insects that alter myrosinase‐catalysed hydrolysis of glucosinolates

The defensive function of the glucosinolate-myrosinase system in plants of the order Capparales results from the formation of isothiocyanates when glucosinolates are hydrolysed by myrosinases upon tissue damage. In some glucosinolate-containing plant species, as well as in the insect herbivore Pieris rapae, protein factors alter the outcome of myrosinase-catalysed glucosinolate hydrolysis, leading to the formation of products other than isothiocyanates. To date, two such proteins have been identified at the molecular level, the epithiospecifier protein (ESP) from Arabidopsis thaliana and the nitrile-specifier protein (NSP) from P. rapae. These proteins share no sequence similarity although they both promote the formation of nitriles. To understand the biochemical bases of nitrile formation, we compared some of the properties of these proteins using purified preparations. We show that both proteins appear to be true enzymes rather than allosteric cofactors of myrosinases, based on their substrate and product specificities and the fact that the proportion of glucosinolates hydrolysed to nitriles does not remain constant when myrosinase activity varies. No stable association between ESP and myrosinase could be demonstrated during affinity chromatography, nevertheless some proximity of ESP to myrosinase is required for epithionitrile formation to occur, as evidenced by the lack of ESP activity when it was spatially separated from myrosinase in a dialysis chamber. The significant difference in substrate- and product specificities between A. thaliana ESP and P. rapae NSP is consonant with their different ecological functions. Furthermore, ESP and NSP differ remarkably in their requirements for metal ion cofactors. We found no indications of the involvement of a free radical mechanism in epithionitrile formation by ESP as suggested in earlier reports.

Allelopathic Effect of Glucosinolate-containing Plant Green Manure on Pythium sp. and Total Fungal Population in Soil

Two Brassicaceae (Iberis amara L. selection ISCI14 and Rapistrum rugosum All. selection ISCI4) and a Capparidacea (Cleome hassleriana L. selection ISCI2) possessing glucosinolates whose degradation products exhibit high fungitoxic activity in vitro were assayed as biocidal plants in a green manure simulation. The trials were carried out in pots, using aboveground fresh plant tissues incorporated at a realistic field rate into soil naturally infected by Pythium sp. The effect of these plant tissues on total fungal populations and Pythium sp. were compared with Crambe abyssinica H. cv. Mario, a Brassicacea containing glucosinolates whose degradation products exhibit low fungitoxic activity in vitro, and a plant (Helianthus annuus L.) not containing glucosinolates. All green manure treatments induced increases in total fungi over a 10-week period, showing an enhanced microflora level compared with untreated soil. Pythium sp. was strongly suppressed by the C. hassleriana, I. amara, and R. rugosum selections, while sunflower and crambe treatments increased Pythium sp. in a manner similar to that observed for total fungal population. These findings indicate that the green manures assayed suppress Pythium sp. and also induced an increase in total soil microbial activity.

Sequestration of host plant glucosinolates in the defensive hemolymph of the sawfly Athalia rosae.

Interactions between insects and glucosinolate-containing plant species have been investigated for a long time. Although the glucosinolate-myrosinase system is believed to act as a defense mechanism against generalist herbivores and fungi, several specialist insects use these secondary metabolites for host plant finding and acceptance and can handle them physiologically. However, sequestration of glucosinolates in specialist herbivores has been less well studied. Larvae of the tumip sawfly Athalia rosae feed on several glucosinolate-containing plant species. When larvae are disturbed by antagonists, they release one or more small droplets of hemolymph from their integument. This "reflex bleeding" is used as a defense mechanism. Specific glucosinolate analysis, by conversion to desulfoglucosinolates and analysis of these by high-performance liquid chromatography coupled to diode array UV spectroscopy and mass spectrometry, revealed that larvae incorporate and concentrate the plant's characteristic glucosinolates from their hosts. Extracts of larvae that were reared on Sinapis alba contained sinalbin, even when the larvae were first starved for 22 hr and, thus, had empty guts. Hemolymph was analyzed from larvae that were reared on either S. alba, Brassica nigra, or Barbarea stricta. Leaves were analyzed from the same plants the larvae had fed on. Sinalbin (from S. alba), sinigrin (B. nigra), or glucobarbarin and glucobrassicin (B. stricta) were present in leaves in concentrations less than 1 micromol/g fresh weight, while the same glucosinolates could be detected in the larvae's hemolymph in concentrations between 10 and 31 micromol/g fresh weight, except that glucobrassicin was present only as a trace. In larval feces, only trace amounts of glucosinolates (sinalbin and sinigrin) could be detected. The glucosinolates were likewise found in freshly emerged adults, showing that the sequestered phytochemicals were transferred through the pupal stage.

Differences in glucosinolate patterns and arbuscular mycorrhizal status of glucosinolate-containing plant species.

Under defined laboratory conditions it was shown that two glucosinolate-containing plant species, Tropaeolum majus and Carica papaya, were colonized by arbuscular mycorrhizal (AM) fungi, whereas it was not possible to detect AM fungal structures in other glucosinolate-containing plants (including several Brassicaceae). Benzylglucosinolate was present in all of the T. majus cultivars and in C. papaya it was the major glucosinolate. 2-Phenylethylglucosinolate was found in most of the non-host plants tested. Its absence in the AM host plants indicates a possible role for the isothiocyanate produced from its myrosinase-catalysed hydrolysis as a general AM inhibitory factor in non-host plants. The results suggest that some of the indole glucosinolates might also be involved in preventing AM formation in some of the species. In all plants tested, both AM hosts and non-hosts, the glucosinolate pattern was altered after inoculation with one of three different AM fungi (Glomus mosseae, Glomus intraradices and Gigaspora rosea), indicating signals between AM fungi and plants even before root colonization. The glucosinolate induction was not specifically dependent on the AM fungus. A time-course study in T. majus showed that glucosinolate induction was present during all stages of mycorrhizal colonization.

Hydrolysis products of glucosinolates in Brassica napus tissues as inhibitors of seed germination

Enzymatic hydrolysis of glucosinolates, a class of compounds found in Brassica species, results in a number of products with potential to inhibit seed germination. To investigate the impact of both volatile and water-soluble allelochemicals, germination bioassays using Lactuca sativa seeds were conducted with root and combined leaf and stem tissues of Brassica napus. Tissues in which glucosinolates were hydrolyzed to remove volatile glucosinolate degradation products were compared with intact tissues and water controls. Only tissues containing glucosinolates produced volatiles that inhibited germination. Volatiles were trapped and identified using GC-MS. Volatiles produced in greater quanitity from intact tissues than from tissues without glucosinolates were almost exclusively glucosinolate hydrolysis products. Water-soluble components also inhibited germination. Chemical analysis of extracts confirmed the presence of glucosinolate hydrolysis products, but indicated the involvement of additional allelochemicals, especially in leaf and stem tissues. Results support the proposal that glucosinolate-containing plant tissues may contribute to reductions in synthetic pesticide use if weed seeds are targeted.

Glucosinolate-containing plant tissues as bioherbicides

ADVERTISEMENT RETURN TO ISSUEPREVArticleNEXTGlucosinolate-containing plant tissues as bioherbicidesPaul D. Brown and Matthew J. MorraCite this: J. Agric. Food Chem. 1995, 43, 12, 3070–3074Publication Date (Print):December 1, 1995Publication History Published online1 May 2002Published inissue 1 December 1995https://doi.org/10.1021/jf00060a015RIGHTS & PERMISSIONSArticle Views951Altmetric-Citations135LEARN ABOUT THESE METRICSArticle Views are the COUNTER-compliant sum of full text article downloads since November 2008 (both PDF and HTML) across all institutions and individuals. These metrics are regularly updated to reflect usage leading up to the last few days.Citations are the number of other articles citing this article, calculated by Crossref and updated daily. Find more information about Crossref citation counts.The Altmetric Attention Score is a quantitative measure of the attention that a research article has received online. Clicking on the donut icon will load a page at altmetric.com with additional details about the score and the social media presence for the given article. Find more information on the Altmetric Attention Score and how the score is calculated. Share Add toView InAdd Full Text with ReferenceAdd Description ExportRISCitationCitation and abstractCitation and referencesMore Options Share onFacebookTwitterWechatLinked InReddit PDF (2 MB) Get e-Alerts Get e-Alerts

Toxicity of allyl isothiocyanate-amended soil toLimonius californicus (Mann.) (Coleoptera: Elateridae) wireworms

Acute toxicity of soil amended with allyl isothiocyanate (AITC) to three size classes ofLimonius californicus (Mann.) wireworms was determined in the laboratory. Wireworms were exposed to AITC at initial concentrations of 120-300 nmol/g soil for one day. During this time, extractable AITC concentrations decreased by 66 to 93 %. Probit analysis estimated LC50 values of 238 and 226 nmol/g soil at one day posttreatment for medium and large wireworms, respectively. For small wireworms, LC50 values decreased from 211 to 157 nmol/g soil during 1-137 days posttreatment. Sublethal concentrations of AITC significantly reduced feeding activity of treated wireworms at three posttreatment times and over the entire 137 days. Wireworm weight was not significantly affected by AITC. The potential exists to use glucosinolate-containing plant tissue as an isothiocyanate (ITC) source to reduce crop damage caused byL. californicus wireworms.