The phytohormone methyl jasmonate as an activator of induced resistance against the necrotroph Alternaria porri f. sp. solani in tomato plants

Abstract

Exogenous application of 0.1 mM methyl jasmonate (MeJA) throughout seed soaking or fumigation of seedlings could induce resistance against the necrotrophic fungus Alternaria porri f. sp. solani in tomato. MeJA applied at 0.01, 0.1, and 1 mM was found to reduce spore germination and mycelial growth in vitro. This compound at 0.01 and 0.1 mM did not cause toxic responses in the tomato plants; however, ethylene production by seedlings was observed to increase after using of all concentrations. A significant increase in the levels of defense markers such as total phenolics, anthocyanins, and phenylalanine ammonia-lyase activity, in response to exogenous MeJA, was observed. Pretreatment of tomato by soaking the seeds in MeJA or treating them with gaseous MeJA efficiently reduced the development of disease caused by Alternaria only when MeJA was applied at 0.1 mM concentration. Seed priming is an easier method of resistance induction than exposure to gaseous MeJA.     

Influence of calcium, potassium, and magnesium on Cornus florida L. density and resistance to dogwood anthracnose

Dogwood anthracnose, a major disease of Cornus florida L., has caused heavy mortality of C. florida in eastern United States forests. Disease severity and rate of infection have been shown to vary with several environmental factors, but the link between soil cation availability and anthracnose has not been examined. We hypothesized that soil cation availability, particularly calcium (Ca), potassium (K), and magnesium (Mg), would influence dogwood survival from anthracnose. In forested stands, positive correlations between soil Ca, K, and Mg saturation and C. florida stem density and basal area were found. The effect of these cations at four levels (0, 50, 100, and 200%) of a standard nursery fertilization rate on C. florida seedling survival and resistance to dogwood anthracnose was tested. Although most of the seedlings died after one season of exposure to dogwood anthracnose, seedlings that had lower inputs of Ca and K cations showed higher levels of disease severity sooner than seedlings in other treatments, suggesting that these nutrients play a role in C. florida survival from anthracnose. Magnesium treatment levels did not appear to have an effect on C. florida disease severity or mortality.     

Use of the maize transposonsActivator andDissociation to show that phosphinothricin and spectinomycin resistance genes act non-cell-autonomously in tobacco and tomato seedlings

Cell-autonomous genes have been used to monitor the excision of both endogenous transposons in maize andAntirrhinum, and transposons introduced into transgenic plants. In tobacco andArabidopsis, the streptomycin phosphotransferase (SPT) gene reveals somatic excision of the maize transposonActivator (Ac) as green sectors on a white background in cotyledons of seedlings germinated in the presence of streptomycin. Cotyledons of tomato seedlings germinated on streptomycin-containing medium do not bleach, suggesting that a different assay for transposon excision in tomato is desirable. We have tested the use of the spectinomycin resistance (SPEC) gene (aadA) and a Basta resistance (BAR) gene (phosphinothricin acetyltransferase, or PAT) for monitoring somatic excision ofAc in tobacco and tomato. Both genetic and molecular studies demonstrate that genotypically variegated individuals that carry clones of cells from whichAc orDs have excised from either SPEC or BAR genes, can be phenotypically completely resistant to the corresponding antibiotic. This demonstrates that these genes act non-cell-autonomously, in contrast to the SPT gene in tobacco. Possible reasons for this difference are discussed.     

The influence of pH and external H+ concentration on caesium toxicity and accumulation inEscherichia coli andBacillus subtilis

Summary Toxicity screening ofEscherichia coli NCIB 9484 andBacillus subtilis 007, NCIB 168 and NCIB 1650 has shown Cs⁺ to be the most toxic Group 1 metal cation. However, toxicity and accumulation of Cs⁺ by the bacteria was affected by two main external factors; pH and the presence of other monovalent cations, particularly K⁺. Over the pH range 6–9 bothE. coli andB. subtilis showed increasing sensitivity towards caesium as the pH was raised. The presence of K⁺ and Na⁺ in the laboratory media used lowered caesium toxicity and lowered acumulation of the metal. In order to assess accurately Cs⁺ toxicity towards the bacterial strains it was therefore necessary to define the K⁺:Cs⁺ ratio in the external medium. The minimum inhibitory K⁺:Cs⁺ concentration ratio for theBacillus strains tested was in the range 12–13 whileE. coli had a minimum inhibitory K⁺:Cs⁺ concentration ratio of 16.     

Modest calcium increase in tomatoes expressing a variant of <Emphasis Type="Italic">Arabidopsis</Emphasis> cation/H<Superscript>+</Superscript> antiporter

The over-expression of Arabidopsis CAX1 and CAX2 causes transgenic tomato plants to reveal severe Ca²⁺ deficiency-like symptoms such as tip-burn and/or blossom end rot, despite there being sufficient Ca²⁺ in each plant part. To correct the symptoms and to moderately enhance the calcium level, a worldwide vegetable tomato was genetically engineered using a modified Arabidopsis cation/H⁺ antiporter sCAX2A, a mutant form of Arabidopsis CAX2. Compared with the wild-type, the sCAX2A-expressing tomato plants demonstrated elevated Ca²⁺ levels in the fruits with almost no changes in the levels of Mn²⁺, Cu²⁺, and Fe²⁺. Moreover, expression of sCAX2A in tomato plants did not show any significant alterations in their morphological phenotypes. Unlike 35S::sCAX1 construct, sCAX2A antiporter gene driven by 35S promoter can be a valuable tool for enriching Ca²⁺ contents in the tomato fruit without additional accumulation of the undesirable cations.     

番茄内生菌的分离鉴定及菌株FQ-G3抗病促生特性

由灰葡萄孢(Botrytis cinerea)引起的灰霉病是番茄生产中最重要的病害之一,当前使用的杀菌剂因药物残留、病原菌抗药性及食品安全等原因逐渐受到限制。因此,利用拮抗微生物的生物防治逐渐成为灰霉病防控的有效策略。【目的】从番茄植株体内筛选具有抗病促生特性内生菌株并对其生防潜力进行评估,为开发番茄灰霉病生物防治新策略提供理论依据。【方法】采用组织分离法在番茄植株不同部位分离出内生细菌、真菌,结合16SrRNA和ITS序列分析,对候选菌株进行初步鉴定;通过菌株对峙培养、果实离体接种筛选对灰葡萄孢具有拮抗活性的内生菌;进一步测定菌株分泌生长素、嗜铁素的能力及其对拟南芥和番茄幼苗生长的促生特性。【结果】从番茄植株不同部位共分离出72株内生细菌和31株内生真菌,通过平板对峙法筛选出1株对多种病原菌具有较好抑菌活性的内生细菌FQ-G3,分子鉴定为Bacillus velezensis。FQ-G3对灰葡萄孢抑菌率达80.93%,并显著抑制灰葡萄孢在番茄果实上的扩展。该菌株能够分泌生长素、蛋白酶和嗜铁素,且对拟南芥、番茄幼苗具有明显的促生效果。【结论】本研究表明分离自番茄植株的内生菌FQ-G3具...     

The Effect of Cations on the Amidase Activity of Human Tissue Kallikrein: 1-Linear Competitive Inhibition by Sodium,Potassium, Calcium and Magnesium. 2-Linear Mixed Inhibition by Aluminium

Hydrolysis of D-valyl-L-leucyl-L-arginine p-nitroanilide by human tissue kallikrein (hK1) was studied in the absence and in the presence of increasing concentrations of the following chloride salts: sodium, potassium, calcium, magnesium and aluminium. The data indicate that the inhibition of hK1 by sodium, potassium, calcium and magnesium is linear competitive and that divalent cations are more potent inhibitors of hK1 than univalent cations. However the inhibition of hK1 by aluminium cation is linear mixed, with the cation being able to bind to both the free enzyme and the ES complex. This cation was the best hK1 inhibitor. Aluminium is not a physiological cation, but is a known neurotoxicant for animals and humans. The neurotoxic actions of aluminium may relate to neuro-degenerative diseases.     

Azione esercitata dal cianuro sulla crescita e sull'inverdimento di plantule di Picea abies (L.) Karsten e di Larix decidua Miller

Abstract

Action of cyanide on growth and greening of seedlings of Picea abies and Larix decidua.—The effects of cyanide on growth and greening of dark germinated seedlings of Picea abies and Larix decidua are studied. While growth of Picea seedlings is scarcely inhibited, the effect of the inhibitor on Larix seedlings is very drastic. Greening of Picea seedlings is never inhibited while, at stronger concentrations, cyanide weakly inhibits greening of Larix seedlings. When seedlings are treated with cyanide and salicylhydrossamic acid, an inhibitor of mithocondrial respiration cyanide non-sensible, growth is inhibited more strongly than when cyanide only is used and greening too is inhibited. Cyanide is shown to be able to reach megagametophyte and embryo and to be incorporated steadily.     

Role of EDTA in a simple method for determining toxicity using a bacterial indicator organism

A method to determine toxicity using a bacterium as the indicator organism previously developed (Botsford 1998) perceives most divalent cations as being toxic. Mercury is perceived as the most toxic, followed by cadmium, zinc and copper. It was found that adding 2.5 m EDTA to the reaction would relieve the toxicity of the 15 divalent cations tested. This effect does not appear to be simple chelation. One micromolar EDTA eliminated the toxicity of 1.6 m calcium or 0.006 m mercury. Thirty-six chemicals were tested for their toxicity in the presence and absence of 2.5 m EDTA and 25 ppm calcium. Twenty-one were less toxic and two of these, p-aminobenzoic acid and tetrachloroethylene would no longer appear to be toxic according to the assay when these additions were present. Six chemicals had the same toxicity with and without the additions. Nine chemicals were more toxic when the EDTA and calcium were present. This experiment was repeated with six chemicals and ten times the EDTA concentration and ten times the calcium concentration. The toxicity with 10× was compared with the toxicity with 1× the additions. The toxicity of 4 of the six chemicals changed with the higher concentration of EDTA and calcium when the absorbancy values observed in samples with the lower levels were compared with samples with the higher levels. Obviously before EDTA can be added to mitigate the toxicity of divalent cations, it must be determined how much EDTA is required to eliminate the toxicity by the ions present in the sample. Alternatively, if the nature of the contaminating organic chemical is known, it can be determined what the effect of EDTA and the divalent cation present is on the apparent toxicity of the compound.     

Effects du NaCl sur les Capacités Oxydases durant la Germination de Semences de Plantes Sensible ou Tolerante au Sel

IAA oxidase, catalase and peroxidase activities of seedlings (the first day of radicle emergence) or organs of young plants (the fifth day of radicle emergence) germinated in NaCl media (0, 25, 50, 100 or 150 mM) were measured. Three species with successively lower thresholds of salinity sensitivity were measured: tomato, radish and red cabbage. NaCl sensitivity of the above enzymes was also analysed in extracts from control seedlings or plants incubated in media with different NaCl concentrations. No relation was found between enzyme activities and protein levels or Na contents of young plants; on the contrary, a good relation was observed between enzyme activity and Na contents of seedlings. Oxidase activities have similar NaCl sensitivity in the three seedlings. Comparing the enzyme activity and sensitivity with NaCl tolerance of the species, the consequences of the oxidase activities in young plants germinated in NaCl media are discussed.      

FIELD GERMINATION AND SEEDLING GROWTH OF CH AND CL PROGENY OF IMPATIENS CAPENSIS (BALSAMINACEAE)

The germination behavior and early growth of chasmogamous (CH) and cleistogamous (CL) progeny of Impatiens capensis were investigated in two eastern Nebraska stands. Field germination of families of buried seeds was scored in mid-April. In one stand a significantly higher proportion of CL seeds germinated than CH seeds while no significant differences were observed in the second stand. Among-family variation in germination rates was significant in both stands. The time course of field germination was similar for CL and CH seeds. A sample of the germinated CL and CH seedlings, matched in size, was planted individually in pots and grown in the greenhouse for five wk. Early growth of CL and CH seedlings was similar. In addition, CL seedlings were as variable as CH seedlings for four morphological traits. Overall, a large fitness advantage for CH progeny was not detected in the germination and early growth of Impatiens capensis seedlings.     

The effect of calcium on the production of ethylene and abscisic acid by fungus Fusarium culmorum and by wheat seedlings infected with that pathogen

Wheat seedlings infected and non-infected with Fusarium culmorum were grown on mediums with different content of calcium (0, 2, 4, 8 mM). It was found that the higher the content of calcium in the medium, the greater the amounts of ethylene produced in both non-infected and infected wheat seedlings, whereas the level of ABA in their tissues was decreased. Taking into consideration the fact that ethylene inhibits, whereas ABA stimulates the growth and development of Fusarium culmorum, it may be assumed that the production of ethylene increased under the influence of calcium and the decreased level of ABA in wheat seedlings causes the reduction growth and development of pathogen and as a result it lowers the degree of infection of wheat seedlings by this fungus. Thus, on the base of the obtained results it may be concluded that the inhibiting influence of calcium on injurying the wheat seedlings by Fusarium culmorum may be caused by the influence of this cation on the hormone balance in the plant.     

Response of plants to calcium concentration in flowing solution culture with chloride or sulphate as the counter-ion

Summary Solution calcium concentrations required for the growth of a range of plant species, including both monocotyledons and dicotyledons, were determined in two experiments in which plants were grown in flowing solution culture at constantly maintained calcium concentrations ranging from 0.5 to 3000 μM. Calcium chloride was used as the calcium source in the first experiment, calcium sulphate was used in the second. At calcium concentrations of 10 μM and below, all species developed calcium deficiency symptoms. The severity of the deficiency was more pronounced in the dicotyledons than in the monocotyledons. However, cassava was much more tolerant than all other dicotyledons and equally as tolerant as rice, the most tolerant monocotyledon. Solution calcium concentrations required for 90% of maximum yield were generally lower for monocotyledons (3 to 20 μM) than for dicotyledons (7 to 720μM) when calcium chloride was used as the calcium source. When calcium sulphate was used, 7 out of 11 species, including 3 monocotyledons, required external calcium concentrations of 1200 μM and above. The results are discussed in relation to effects of solution composition and the choice of counter-ions on plant response to calcium and other macronutrient cations. It is concluded that yield depressions due to toxicity of excesses of chloride, and possibly other counter-ions, can lead to serious underestimation of limiting external cation concentrations for plant growth.     

Investigation on the Fusarium induced stress protein (FISP) in wheat: Immunolocalisation in root cells

Abstract

Fusarium induced-stress-protein (FISP) of ～51 kDa molecular mass was detected in seven day old germinated wheat (Triticum aestivum var Sonalika) seedlings infected with F. oxysporum for a period of seven days. This particular stress protein (FISP) of ～51 kDa was over-expressed in the case of Fusarium infected seedlings compared to the untreated seedlings where the presence of this protein was insignificant. Localisation of this ～51 kDa protein in root tissue by anti-CSAP (Cadmium Stress Associated Protein) antiserum showed a significantly higher number of gold particles in the case of Fusarium infected root tissue compared to the untreated control. A unique type of organised localisation of FISP around the plasma membrane and outer vacuolar membrane suggests its defensive role against Fusarium infection that might be a general stress protein against biotic and abiotic stresses.     

Short communication. Identification of developmentally-specific markers in germinating and haustorial stages of Striga hermonthica (Del.) Benth. seedlings

Developmentally-specific markers have been identified in the germinating and haustorial stages of Striga hermonthica seedlings. Four water-soluble proteins, preferentially expressed in germinated seedlings, were microsequenced. An haustorial-specific cDNA clone was isolated by differential screening. Tissue specificity for this clone was assessed by Northern blot hybridization analysis.Key words: Striga hermonthica (Del.) Benth, microsequencing, lipid transfer protein, superoxide dismutase.      

Control of Fusarium wilt of tomato caused by Fusarium oxysporum f. sp. lycopersici using leaf extract of Piper betle L.: a preliminary study

The main objective of this study was to evaluate the effectiveness of crude chloroform extract of Piper betle L. (PbC) in controlling Fusarium wilt of tomato (Lycopersicon esculentum) caused by Fusarium oxysporum f. sp. lycopersici. It was observed that 1% (w/w) amendment of the PbC in soil was more efficient in reducing the Fusarium population in soil than carbendazim and the combined amendment of carbendazim and PbC. Fusarium wilt control studies were carried out in a greenhouse. Variation in different parameters like shoot growth, root growth and mean fresh weights of tomato seedlings in all the treatments were recorded. Accumulation of total phenolics was also studied from the root tissues of tomato. Higher accumulation of total phenolics was observed in the Fusarium-infested plants as compared to that of healthy control and PbC-treated plants. Moreover, it was observed that the extract could reduce the symptoms and disease development. Electron microscopy studies were also done to observe the Fusarium infestation in the vascular bundles and to show the accumulation of total phenolics in the vacuoles of root tissue.     

Azione esercitata da alcuni inibitori respiratori sulla crescita e sull'inverdimento di plantule di Picea abies e di Larix decidua

Abstract

Action of some respiratory inhibitors on growth and greening of seedlings of Picea and Larix.—The effects of sodium azide, arseniate, jodoacetate, fluoride and 2,4-dinitrophenol on growth and greening of dark germinated seedlings of Picea abies and Larix decidua were studied.

All the inhibitors showed a negative influence on growth (determined as fresh weight) of Picea seedlings and even more remarkably on Larix seedlings.

Greening was significantly inferior to that of control plants only for groups of Picea seedlings treated with sodium azide, at all the concentrations used, and for groups treated with fluoride and arseniate at the highest concentrations.

All the groups of Larix seedlings, instead, treated with inhibitors, with the exception of the ones treated with sodium fluoride at the lowest concentrations, presented a chlorophyll content inferior to that of control groups.

From the above results the different influence of inhibitors on Picea and on Larix seedlings might be due to the different metabolic pathways through which the pigment synthesis in the two species under study takes place.     

Bacillus halotolerans strain LYSX1-induced systemic resistance against the root-knot nematode Meloidogyne javanica in tomato

Purpose

Determination of the nematicidal potential and mode of action of bacteria isolated from tobacco rhizosphere soil against the root-knot nematode Meloidogyne javanica in tomato plants.

Methods

Antagonistic bacteria were isolated from rhizosphere soil of tobacco infested with root-knot nematodes. Culture filtrate was used to examine nematicidal activity and ovicidal action of bacterial strains. Biocontrol of M. javanica and growth of treated tomato plants were assessed in pot experiments. To clarify whether secondary metabolites of bacteria in tomato roots induced systemic resistance to M. javanica, bacterial culture supernatants and second-stage juvenile nematodes were applied to spatially separated tomato roots using a split-root system. Bacterial strains were identified by 16S rDNA and gyrB gene sequencing and phylogenetic analysis.

Results

Of the 15 bacterial strains isolated, four (LYSX1, LYSX2, LYSX3, and LYSX4) demonstrated nematicidal activity against second-stage juveniles of M. javanica, and strain LYSX1 showed the greatest antagonistic activity; there was dose-dependent variability in nematicidal activity and inhibition of egg mass hatching by strain LYSX1. In vivo application of LYSX1 to tomato seedlings decreased the number of egg masses and galls and increased the root and shoot fresh weight. Treatment of half of the split-root system with LYSX1 reduced nematode penetration to the other half by 41.64%. Strain LYSX1 was identified as Bacillus halotolerans.

Conclusion

Bacillus halotolerans LYSX1 is a potential microbe for the sustainable biocontrol of root-knot nematodes through induced systemic resistance in tomato.

     

Role of Bacillus amyloliquefaciens Y1 in the control of Fusarium wilt disease and growth promotion of tomato

The objective of this study was to examine the effects of Bacillus amyloliquefaciens Y1 on the control of Fusarium wilt disease and subsequent improvement in the growth of tomato plants. The Y1 strain strongly inhibited Fusarium oxysporum f. sp. lycopersici in vitro and also produced indole-3-acetic acid (IAA) in both the presence and absence of tryptophan. Over 96% of tomato seeds germinated when treated with either water, tryptone soy broth, or Y1 cultures, whereas root (5.40?cm) and shoot (5.15?cm) lengths were greatest in tomato seedlings treated with Y1 cultures that lacked tryptophan. Three experimental treatments – Black White medium (BW), BW medium with a commercial fungicide (BW?+?F), and Y1 culture inoculated in BW medium (Y1) – were applied to control Fusarium wilt disease under in vivo conditions. Application of Y1 culture and BW?+?F led to significantly lower disease incidence than did BW; moreover, shoot length and fresh and dry weight of both roots and shoots were greater in plants treated with Y1 than in plants treated with either BW or BW?+?F. A similar trend was observed for chitinase and β-1,3-glucanase activities in roots and leaves of tomato plants in all treatment groups over most of the experimental period. Finally, the presence of Y1 in the rhizospheric soils of Y1-treated plants resulted in a significant reduction in the populations of other bacteria. The results of our study demonstrated the effectiveness of Y1 not only in the control of Fusarium wilt disease but also for the enhancement of plant growth in cultivated tomato.     

GABA Requires Nitric Oxide for Alleviating Arsenate Stress in Tomato and Brinjal Seedlings

In recent years, gamma-aminobutyric acid (GABA) and nitric oxide (NO) have drawn much attention in reducing/alleviating metal toxicity in plants. Arsenic is a potentially toxic metal that imposes severe toxic effects on crop plants. Therefore, the present study was an attempt to explore the role of GABA and NO in the amelioration of arsenate As(V) toxicity in tomato and brinjal seedlings. Arsenate toxicity reduced fresh mass about by 17% in tomato and 25% in brinjal seedlings, increased the level of reactive oxygen species, antioxidant enzymes (except glutathione-S-transferase), and altered photosynthesis. However, exogenously applied GABA and sodium nitroprusside (SNP, a NO donor) improved the growth of both vegetables and reduced As(V) toxicity. Interestingly, As(V) toxicity was further increased on applying biosynthetic inhibitor of NO even in the presence of GABA, and under similar conditions addition of NO donor rescued the negative effect of biosynthetic inhibitor (L-NAME) of NO. This indicates that NO is crucial in GABA-mediated alleviation of As(V) toxicity in tomato and brinjal seedlings. Besides this, results revealed that c-PTIO (a NO scavenger) addition reversed the alleviatory effect of SNP also suggesting that NO is important in curtailing As(V) toxicity. Overall, the results indicated that NO is necessary for mitigation of As(V) stress by GABA in two studied vegetable crops but NO itself does not require GABA for the same.