Striga seed-germination activity of root exudates and compounds present in stems of Striga host and nonhost (trap crop) plants is reduced due to root colonization by arbuscular mycorrhizal fungi

Root colonization by arbuscular mycorrhizal (AM) fungi reduces stimulation of seed germination of the plant parasite Striga (Orobanchaceae). This reduction can affect not only host plants for Striga, resulting in a lower parasite incidence, but also false hosts or trap crops, which induce suicidal Striga seed germination, thereby diminishing their effectiveness. In order to better understand these AM-induced effects, we tested the influence of root colonization by different AM fungi on the seed-germination activity of root exudates of the Striga hermonthica nonhost plants cowpea and cotton on S. hermonthica. We also tested the effect of AM fungi on the seed-germination activity of the Striga gesnerioides host plant cowpea on S. gesnerioides. Moreover, we studied whether mycorrhization affects the transport of seed-germination activity to above-ground plant parts. Mycorrhization not only resulted in a lower seed germination of S. gesnerioides in the presence of root exudates of the S. gesnerioides host cowpea but also seed germination of S. hermonthica was also lower in the presence of root exudates of the S. hermonthica nonhosts cowpea and cotton. Downregulation of the Striga seed-germination activity occurs not only in root exudates upon root colonization by different AM fungi but also in the compounds produced by stems. The lowered seed-germination activity does not appear to depend on the presence of seed germination inhibitors in the root exudates of mycorrhizal plants. The implication for Striga control in the field is discussed.     

Development of a model system to assess the impact of genetically modified corn and aubergine plants on arbuscular mycorrhizal fungi

We developed an experimental model system to monitor the impact of generically modified (GM) plants on arbuscular mycorrhizal (AM) fungi, a group of non-target soil microorganisms, fundamental for soil fertility and plant nutrition. The system allowed us to study the effects of root exudates of both commercial Bt corn and aubergine plants expressing Dm-AMP1 defensin on different stages of the life cycle of the AM fungal species G. mosseae. Root exudates of Bt 176 corn significantly reduced pre-symbiotic hyphal growth, compared to Bt 11 and non-transgenic plants. No differences were found in mycelial growth in the presence of Dm-AMP1 and control plant root exudates. Differential hyphal morphogenesis occurred irrespective of the plant line, suggesting that both exuded Bt toxin and defensin do not interfere with fungal host recognition mechanisms. Bt 176 affected the regular development of appressoria, 36% of which failed to produce viable infection pegs. Our experimental model system represents an easy assay for testing the impact of GM plants on non-target soil-borne AM fungi.     

Production of Strigolactones by Arabidopsis thaliana responsible for Orobanche aegyptiaca seed germination

The germination stimulants produced by Arabidopsis thaliana, a host of root parasitic plants Orobanche spp. but not of arbuscular mycorrhizal (AM) fungi were examined. Root exudates from the hydroponically grown A. thaliana plants were subjected to reverse phase high performance liquid chromatography (HPLC) and retention times of germination stimulants inducing O. aegyptiaca seed germination were compared with those of strigolactone standards. In addition, the root exudates were analyzed by using HPLC linked with tandem mass spectrometry (LC/MS/MS). A. thaliana was found to exude at least three different germination stimulants of which one was identified as orobanchol. This is the first report of strigolactone production by a non-mycotrophic plant. These results together with recent knowledge imply that strigolactones have other unrevealed functions in plant growth and development.     

Alterations in Root Exudation of Intercropped Tomato Mediated by the Arbuscular Mycorrhizal Fungus Glomus mosseae and the Soilborne Pathogen Fusarium oxysporum f.sp. lycopersici

Arbuscular mycorrhizal fungi (AMF) can control soilborne diseases such as Fusarium oxysporum f.sp. lycopersici (Fol). Root exudates play an important role in plant–microbe interactions in the rhizosphere, especially, in the initial phase of these interactions. In this work, we focus on (i) elucidating dynamics in root exudation of Solanum lycopersicum L. in an intercropping system due to AMF and/or Fol; (ii) its effect on Fol development in vitro; and (iii) the testing of the root exudate compounds identified in the chromatographic analyses in terms of effects on fungal growth in in vitro assays. GC‐MS analyses revealed an AMF‐dependent increase in sugars and decrease in organic acids, mainly glucose and malate. In the HPLC analyses, an increase in chlorogenic acid was evident in the combined treatment of AMF and Fol, which is to our knowledge the first report about an increase in chlorogenic acid in root exudates of AM plants challenged with Fol compared with plants inoculated with AMF only, clearly indicating changes in root exudation due to AMF and Fol. Root exudates of AMF tomato plants stimulate the germination rate of Fol, whereas the co‐inoculation of AMF and Fol leads to a reduction in spore germination. In the in vitro assays, citrate and chlorogenic acid could be identified as possible candidates for the reduction in Fol germination rate in the root exudates of the AMF+Fol treatment because they proved inhibition at concentrations naturally occurring in the rhizosphere.     

Evidence for leaf endophyte regulation of root symbionts: effect of <Emphasis Type="Italic">Neotyphodium</Emphasis> endophytes on the pre-infective state of mycorrhizal fungi

Neotyphodium endophytes and arbuscular mycorrhizal (AM) fungi are common constituents of natural grasslands. The plant–endophyte symbiosis can introduce changes in soil conditions that affect the density and activity of different functional groups of soil organisms. In the present work we performed in vitro assays to evaluate the effect of root and endophyte exudates on the pre-infective state of mycorrhizal fungi (Gigaspora margarita and G. rosea). Plant roots of Bromus setifolius from populations of Patagonia, and four strains of Neotyphodium were used to obtain the exudates. Root exudates of infected plants, at a high concentration, significantly increased AMF hyphal branches and length relative to exudates from naturally endophyte free plants. The effect of Neotyphodium endophyte exudates on AMF mycelial length varied depending on strain and the concentration used, suggesting a differential interaction between endophyte and AMF species. AMF hyphal branches were increased by Neotyphodium fungal exudates in both mycorrhizal species. A few previous studies have suggested that Neotyphodium endophytes can reduce mycorrhizal sporulation and colonization of host roots in commonly-cultivated agronomic hosts. In this study we report the opposite effect in B. setifolius. This study reports the direct and positive effect of root exudates from plants in symbiosis with Neotyphodium, on AMF pre-infective state. Further, identical effects were detected using exudates from Neotyphodium endophytes.     

柏木根系分泌物对栾树细根形态及N、P含量的影响

为研究混交过程中柏木根系分泌物对栾树细根生长的影响,以一年生栾树盆栽幼苗为研究对象,通过施加1株、2株、4株、8株4个浓度柏木根系分泌物(分别记为G1、G2、G4、G8)于栾树盆栽中,探讨柏木根系分泌物对栾树幼苗1～5级细根形态及N、P含量的影响。结果表明:(1)栾树细根直径随根序的增加而增大,施加根系分泌物显著减小了1～2级细根的直径(P0.05);细根比根长、比表面积均随根序的增加而减小,施加根系分泌物显著增大了1～2级细根的比根长及比表面积(P0.05);随根系分泌物施加浓度的提高,栾树比根长及比表面积先增大,而直径先减小,然后均趋于平缓波动的态势。(2)栾树细根N、P含量均随根序的增加而减小,而N/P在根序间的变化不显著;施加柏木根系分泌物显著增大了栾树1～2级细根的N、P含量(P0.05),但减小了1～5级细根的N/P;随根系分泌物施加浓度的提高,栾树细根P含量增大,N/P减小,而N含量先增加后呈现平缓变化的趋势。(3)栾树细根N、P含量均与其比根长、比表面积和直径等形态特征之间呈显著的相关关系(P0.05)。研究发现,柏木根系分泌物可改善土壤养分的有效性,从而缓解栾树植株的缺P症状,细根通过调整其形态以提高养分利用效率;柏木根系分泌物主要影响栾树1～2级细根的形态及N、P含量;4株柏木根系分泌物的剂量更有利于栾树根系的生长。     

Root exudates from sunflower (Helianthus annuus L.) show a strong adsorption ability toward Cd(II)

Abstract

Cd(II) adsorption of root exudates from sunflower (Helianthus annuus L.) seedling was investigated by Cd ion-selective electrode, Fourier Transform Infrared spectroscopy, and fluorescence spectroscopy. Root exudates from Helianthus annuus L. had strong adsorption ability toward Cd(II). The adsorption process was pH-dependent and the maximum adsorption capacity, 150.8 mg g^?1, was observed at pH 7.0. Root exudates had pK _a1 at 4.7 for carboxyl and pK _a2 at 9.2 for phenolic, and amino groups. The aliphatic and aromatic (C?H) groups, amide III group, and the C (=O)?O and sulfonate groups were responsible for Cd(II) adsorption. The excitation emission matrix fluorescence spectroscopy showed protein-like substances participated in Cd adsorption and formed strong complexes, with conditional stability constants of 4.70 and 4.32, which is a little lower than that determined by potentiometric methods, 5.13. The strong Cd complexing ability of root exudates implies that root exudates may significantly affect mobility, toxicity, and phytoavailability of Cd. Cd binding of root exudates may be attributed to its interaction with the proteins, polysaccharides, and phenolic compounds in root exudates.     

Mobilization of cadmium and other metals from two soils by root exudates of Zea mays L., Nicotiana tabacum L. and Nicotiana rustica L.

Soluble root exudates were collected from three plants (Nicotiana tabacum L., Nicotiana rustica L. and Zea mays L.), grown under axenic and hydroponic conditions, in order to study their metal-solubilizing ability for Cd and other cations (Cu, Fe, Mn, Ni, Zn). Nicotiana spp. and Zea mays L. root exudates differed markedly in C/N ratio, sugars vs. amino acids ratio and organic acids content. Metals from two soils were extracted with either root exudate solutions, containing equal amounts of organic carbon, or distilled water as control. In the presence or absence of root exudates, the solubility of Fe and Mn was much higher than of the four other metals tested. Root exudates increased the solubilities of Mn and Cu, whereas those of Ni and Zn were not affected. Root exudates of Nicotiana spp. enhanced the solubility of Cd. The extent of Cd extraction by root exudates (N. tabacum L. N. rustica L. Zea mays L.) was similar to the order of Cd bioavailability to these three plants when grown on soil. An increase in Cd solubility in the rhizosphere of apical root zones due to root exudates is likely to be an important cause of the relatively high Cd accumulation in Nicotiana spp.     

Effect of root exudates on the spore germiantion of rhizosphere and rhizoplane mycoflora of chilli (Capsicum annuum L.) cultivars

Summary From root exudates of three cultivars of chilli (Capsicum annuum L.) 12 amino acids and 7 sugars were detected. Methionine, d-1- phenylalanine, citrulline and d-xylose were detected only from the root exudates of resdistant cultivars. The root exudates of resistant variety inhibited spore germination of the pathogen (Fusarium oxysporum f. sp.capsici), but that of susceptible variety enhanced spore germiantion of the same. Spore germiantion of antagonistic fungi (Trichderma viride andAspergillus sydowi) was also influenced by the root exudates of resistant and susceptible varieties, but the influence was different.Spore germiantion of a number of rhizosphere fungi was studied and in general root exudate of susceptible cultivar enhanced spore germiantion of majority of fungi, but spore germination of antagonistic fungi against the pathogen was inhibited. However, root exudate of resistant cultivar stimulated spore germination of antagonistic fungi.     

感染羽茅的香柱菌属内生真菌对丛枝菌根真菌孢子萌发的影响

内生真菌与丛枝菌根(AM)真菌是构成草原生态系统的重要组成部分.内生真菌会抑制其宿主植物的AM真菌侵染率.本研究以感染2种香柱菌属内生真菌[Epichloё gansuensis(Eg)和E. sibirica(Es)]的天然禾草羽茅为供试材料,进行体外纯培养的内生真菌培养滤液、感染内生真菌的羽茅叶片(包括鲜叶和枯叶)浸提液,以及根系分泌物对摩西球囊霉(Gm)和幼套球囊霉(Ge)2种AM真菌孢子萌发影响试验.结果表明: 香柱菌属内生真菌的培养滤液会显著抑制2种AM真菌孢子的萌发,而感染香柱菌属内生真菌的羽茅根系分泌物只对Ge孢子萌发有显著抑制作用,且上述抑制作用与内生真菌种类无关;鲜叶浸提液对Gm和Ge的孢子萌发率均无显著影响,而枯叶浸提液对Ge的孢子萌发有显著抑制作用.在自然生态系统中,香柱菌属内生真菌通常存在于宿主植物体内,可能通过影响宿主植物的根系分泌物来影响AM真菌孢子的萌发.     

Effect of onion (Allium cepa) root exudates on the hyphal growth of Gigaspora margarita

 The effect of root exudates from onions differing in P status on spore germination and hyphal growth of arbuscular mycorrhizal fungi was investigated. Onion (Allium cepa) was grown in solution culture at different phosphorus concentrations (0, 0.1, 1.0, 8.0 and 24.0 mg P l^–1) and root exudates were collected. When spores of the arbuscular mycorrhizal fungus, Gigaspora margarita were incubated with these root exudates, spore germination was only slightly affected but hyphal growth was greatly affected, particularly with exudates from P-deficient plants. This suggests that the P nutrition of host plants influences the composition of root exudates and thereby the hyphal growth of arbuscular mycorrhizal fungi. Accepted: 25 June 1995     

Quantitative and qualitative effects of phosphorus on extracts and exudates of sudangrass roots in relation to vesicular-arbuscular mycorrhiza formation

A comparison was made of water-soluble root exudates and extracts of Sorghum vulgare Pers. grown under two levels of P nutrition. An increase in P nutrition significantly decreased the concentration of carbohydrates, carboxylic acids, and amino acids in exudates, and decreased the concentration of carboxylic acids in extracts. Higher P did not affect the relative proportions of specific carboxylic acids and had little effect on proportions of specific amino acids in both extracts and exudates. Phosphorus amendment resulted in an increase in the relative proportion of arabinose and a decrease in the proportion of fructose in exudates, but did not have a large effect on the proportion of individual sugars in extracts. The proportions of specific carbohydrates, carboxylic acids, and amino acids varied between exudates and extracts. Therefore, the quantity and composition of root extracts may not be a reliable predictor of the availability of substrate for symbiotic vesicular-arbuscular mycorrhizal fungi. Comparisons of the rate of leakage of compounds from roots with the growth rate of vesicular-arbuscular mycorrhizal fungi suggest that the fungus must either be capable of using a variety of organic substrates for growth, or be capable of inducing a much higher rate of movement of specific organic compounds across root cell membranes than occurs through passive exudation as measured in this study.     

Antimicrobial proteins from cowpea root exudates: inhibitory activity against Fusarium oxysporum and purification of a chitinase-like protein

Plants exude a variety of substances through their external surfaces and from germinating seeds, some of which have an inhibitory action against plant pathogens. The aim of this study was the investigation and characterization of defense proteins present in exudates from roots of cowpea seedlings (Vigna unguiculata (L.) Walp.). Root exudates were collected from seedlings that were grown hydroponically in three different media, including, 100 mM sodium acetate buffer pH 4.5, water pH 6.0 and 100 mM sodium phosphate buffer pH 7.5. The proteins from these exudates were analyzed by SDS–PAGE and SDS–Tricine–PAGE and the presence of antimicrobial proteins in the exudates was investigated by immunological and enzymatic assays. Results showed that roots from cowpea seedlings contained -1,3-Glucanases, chitinases and lipid transfer proteins (LTPs), all of which may potentially function as plant defense proteins. Immunolocalization of one of these proteins, chitinase, revealed its presence in the xylem cell wall vessel elements. These exudates also demonstrated an inhibitory effect on the growth of the fungus, Fusarium oxysporum, in vitro. The results suggest that plant roots may exude a variety of proteins that may function to repress the growth of root pathogenic fungi.     

Effect of inoculation on the exudation of amino acids and sugars by berseem (Trifolium alexandrinum) and lucerne (Medicago sativa)

Summary Root exudates of inoculated and uninoculated berseem (Trifolium alexandrinum) and lucerne (Medicago sativa) were examined chromatographically for their amino acid and reducing-sugar contents after 6 and 9 weeks growth in a sand culture experiment.Aspartic acid was the only amino acid detected in the root exudates of both the inoculated and uninoculated legumes, up to 9 weeks growth.Root exudates of inoculated berseem contained more amino acids than those of lucerne. Glutamic acid and fructose are characterised by their consistent presence in the root exudates of inoculated plants and their absence in those of uninoculated ones.The data presented in this paper is taken from the thesis submitted by the senior author to the P.G. School, IARI, New Delhi in 1964 for M. Sc. degree in Microbiology.     

Studies on fungi in the root region

Summary Aqueous extracts of adventitious roots ofAllium sativum L. (garlic), seedling roots ofAllium porrum L. (leek), and both adventitious and seedling roots ofAllium cepa L. (onion), were tested for antibiotic activity against three root-surface fungi,Cylindrocarpon radicicola Wollenw.,Gliocladium roseum (Link) Thom andFusarium culmorum (W. G. Smith) Sacc. by means of two different techniques.With a filter-paper disc technique, root extracts sterilised by membrane-filtration produced zones of inhibition of the test fungi, whereas root extracts sterilised by autoclaving showed no activity. Garlic root extract produced inhibition zones with all the test fungi, whereas extracts of onion adventitious roots and leek seedling roots produced inhibition zones with only one of the test fungi. The extract of onion seedling roots produced no inhibition zones. Root extracts of all theAllium species, when sterilised by membrane filtration, generally inhibited spore-germination of all the test fungi.     

Positive correlation between VAM-induced changes in root exudation and mycorrhizosphere mycoflora

The influence of vesicular-arbuscular mycorrhizal (VAM) fungi on rhizosphere mycoflora was studied together with the possible mechanism involved in this process. Six combinations of VAM fungi and phosphorus fertilizer treatments were applied to Leucaena leucocephala roots and quantitative and qualitative observations were made periodically of the rhizosphere mycoflora and constituents of root exudates. The results obtained indicate that the presence of specific mycoflora in the rhizosphere of mycrorrhizal roots is mediated through root exudates rather than being an outcome of improved P nutrition.     

Root exudates from lentil (Lens culinaris medic) seedlings in relation to wilt disease

Summary Root exudates from healthy and diseased lentil plants (Lens culinaris) have been investigated in relation to the wilt disease caused byFusarium oxysporum f.lentis. In all ten amino acids and five sugars have been detected. The spore germination of the pathogen in root exudates indicated that 21-days root exudate was inhibitory. Glycine and phenylalanine were detected in 21-days exudate and were found to have an inhibitory effect upon the germination of the spores of the pathogen which may partly explain the lesser disease incidence when plants of more than 3 weeks are inoculated with the pathogen.     

Lead complexation behaviour of root exudates of salt marsh plant Salicornia europaea L

Abstract

Root exudates are considered to have an important role in mobility and bioavailability of heavy metals. High molecular weight (HMW) substances are the main components of root exudates, however, knowledge about their interactions with heavy metals is lacking. In the present study, Pb(II) complexation of the HMW fluorescent fractions in root exudates from Salicornia europaea L. was investigated using excitation emission matrix (EEM) fluorescence spectroscopy. Two protein-like fluorescence peaks were identified in the EEM spectrum of root exudates. The fluorescence of both peaks was clearly quenched by Pb(II). The values of conditional stability constants, log K_a, for these two protein-like fluorescence peaks were 4.14 and 3.79. This indicates that the fluorescent substances are strong Pb(II) complexing organic ligands.     

Autotoxic potential of cucurbit crops

Soil sickness is often observed in cucurbit crops such as Citrullus lanatus, Cucumis melo and Cucumis sativus, but not in cucurbit crops such as Cucurbita moschata, Lagenaria leucantha and Luffa cylindrica. Results showed that root aqueous extracts of Citrullus lanatus, Cucumis melo and Cucumis sativus were autotoxic, but those of Cucurbita moschata, Momordica charantia and Luffa cylindrica were less autotoxic to the radicle elongation of respective species. Plant growth of Citrullus lanatus, Cucumis melo and Cucumis sativus were greatly inhibited by autotoxic substances released from powered root tissue at a rate of 1 g per seedling. Root exudates of Citrullus lanatus, Cucumis melo and Cucumis sativus were autotoxic to radicle elongation and seedling growth of respective species. However, root exudates of Citrullus lanatus did not inhibit radicle elongation of Cucurbita ficifolia, which is commonly used as rootstock for the grafting of Citrullus lanatus, Cucumis melo and Cucumis sativus to decrease soil-borne diseases in commercial production. It seems possible to overcome autotoxicity in cucurbit crops by grafting on Cucurbita ficifolia. This revised version was published online in June 2006 with corrections to the Cover Date.     

Assessment of the Role of Chemotaxis and Biofilm Formation as Requirements for Colonization of Roots and Seeds of Soybean Plants by <Emphasis Type="Italic">Bacillus amyloliquefaciens</Emphasis> BNM339

This article correlates colonization with parameters, such as chemotaxis, biofilm formation, and bacterial growth, that are believed to be connected. We show here, by using two varieties of soybean plants that seeds axenically produced exudates, induced a chemotactic response in Bacillus amyloliquefaciens, whereas root exudates did not, even when the exudates, also collected under axenic conditions, were concentrated up to 200-fold. Root exudates did not support bacterial cell division, whereas seed exudates contain compounds that support active cell division and high cell biomass at stationary phase. Seed exudates of the two soybean varieties also induced biofilm formation. B. amyloliquefaciens colonized both seeds and roots, and plant variety significantly affected bacterial root colonization, whereas it did not affect seed colonization. Colonization of roots in B. amyloliquefaciens occurred despite the lack of chemotaxis and growth stimulation by root exudates. The data presented in this article suggest that soybean seed colonization, but not root colonization, by B. amyloliquefaciens is influenced by chemotaxis, growth, and biofilm formation and that this may be caused by qualitative changes of the composition of root exudates.