A profitable function of Ceriporia lacerata HG2011 on nodulation and biological nitrogen fixation of green bean (Phaseolus vulgaris L.).

AIMS: Green bean (Phaseolus vulgaris L.) is a popular vegetable worldwide. The use of beneficial fungi is a simple and effective way to improve the biological nitrogen fixation (BNF) of this leguminous vegetable. METHODS AND RESULTS: A micro-plot was conducted to investigate the enhancement of BNF using 15N natural abundance technology and agronomic performances of green bean caused by wood-rot fungus Ceriporia lacerata HG2011. The results showed the soil for frequently growing green bean featured abundant native rhizobia, and newly inoculated rhizobia may have to compete with them in nodulation and only highly competitive rhizobia can succeed. The addition of C. lacerata HG2011 to the soil increased the population of ammonia oxidizers, nitrifiers, and phosphorus (P)-mobilizing microbes in rhizosphere, accelerated nitrification and P mobilization, creating a favorable soil environment with high P and low ammonia for BNF. Green bean received C. lacerata HG2011 had higher dehydrogenase activity in roots and higher nodulation rate and large nodules. These phenomena implied abundant supplies of adenosine triphosphate, nicotinamide adenine dinucleotide hydrogen, or nicotinamide adenine dinucleotide phosphate hydrogen for BNF in the roots, a large proportion of N2 fixation tissues, and a greater sink for receiving photosynthates. As a result, C. lacerata HG2011 considerably increased the percentage of N derived from the atmosphere, BNF, and plant nutrient uptake (including N, P, and potassium), leading to 15.58%-28.51% of biomass increasment and 9.82%-17.03% of peapod yield increasment along with quality improvement compared with non-fungal application. CONCLUSIONS: C. lacerata HG2011 increased the nodulation and BNF of green bean, accelerated the nutrient uptake (NPK) and therefore improved the yield and peapod quality of green bean. SIGNIFICANCE AND IMPACT OF STUDY: The study demonstrates that C. lacerata HG2011 could be used as a biofertilizer for BNF improvement of legumes.

Differential Virulence Contributions of the Efflux Transporter MexAB-OprM in Pseudomonas syringae Infecting a Variety of Host Plants.

Efflux transporters such as MexAB-OprM contribute to bacterial resistance to diverse antimicrobial compounds. Here, we show that MexB contributes to epiphytic and late-stage apoplastic growth of Pseudomonas syringae strain B728a, as well as lesion formation in common bean (Phaseolus vulgaris). Although a ∆mexB mutant formed fewer lesions after topical application to common bean, these lesions contain the same number of cells (105 to 107 cells) as those caused by the wild-type strain. The internalized population size of both the wild-type and the ∆mexB mutant within small samples of surface-sterilized asymptomatic portions of leaves varied from undetectably low to as high as 105 cells/cm2. Localized bacterial populations within individual lesions consistently exceeded 105 cells/cm2. Strain B728a was capable of moderate to extensive apoplastic growth in diverse host plants, including lima bean (P. lunatus), fava bean (Vicia faba), pepper (Capsicum annuum), Nicotiana benthamiana, sunflower (Helianthus annuus), and tomato (Solanum lycopersicum), but MexB was not required for growth in a subset of these plant species. A model is proposed that MexB provides resistance to as-yet-unidentified antimicrobials that differ between plant species. [Formula: see text] Copyright © 2022 The Author(s). This is an open access article distributed under the CC BY 4.0 International license.

Agrobacterium leguminum sp. nov., isolated from nodules of Phaseolus vulgaris in Spain.

Two endophytic strains, coded MOVP5T and MOPV6, were isolated from nodules of Phaseolus vulgaris plants grown on agricultural soil in Southeastern Spain, and were characterized through a polyphasic taxonomy approach. Their 16S rRNA gene sequences showed 99.3 and 99.4 %, 98.9 and 99.6 %, and 99.0 and 98.7% similarity to 'A. deltaense' YIC 4121T, A. radiobacter LGM 140T, and A. pusense NRCPB10T, respectively. Multilocus sequence analysis based on sequences of recA and atpD genes suggested that these two strains could represent a new Agrobacterium species with less than 96.5 % similarity to their closest relatives. PCR amplification of the telA gene, involved in synthesis of protelomerase, confirmed the affiliation of strains MOPV5T and MOPV6 to the genus Agrobacterium. Whole genome average nucleotide identity and digital DNA-DNA hybridization average values were less than 95.1 and 66.7 %, respectively, with respect to its closest related species. Major fatty acids in strain MOPV5T were C18 : 1 ω7c/C18 : 1 ω6c in summed feature 8, C19 : 0 cyclo ω8c, C16 : 0 and C16 : 0 3-OH. Colonies were small to medium, pearl-white coloured on YMA at 28 °C and growth was observed at 10-42 °C, pH 5.0-10.0 and with 0.0-0.5 % (w/v) NaCl. The DNA G+C content was 59.9 mol%. These two strains differ from all other genomovars of Agrobacterium found so far, including those that have not yet given a Latin name. The combined genotypic, phenotypic and chemotaxonomic data support the classification of strain MOPV5T as representing a novel species of Agrobacterium, for which the name Agrobacterium leguminum sp. nov. is proposed. The type strain is MOPV5T (=CECT 30096T=LMG 31779T).

Activation of defense response in common bean against stem rot disease triggered by Trichoderma erinaceum and Trichoderma viride.

White mold and stem rot is a common disease of Phaseolus vulgaris caused by Sclerotinia sclerotiorum. Biological control is a promising alternative for the control of this disease. In the present study, two Trichoderma spp., T. erinaceum and T. viride, and the consortium of both were evaluated as biocontrol agents against sclerotinia stem rot disease. The results revealed that T. erinaceum (NAIMCC-F-02171) and T. viride (NAIMCC-F-02500) when applied alone, significantly suppressed the infection rate of S. sclerotiorum and increased the rate of survival of plants by 74.5%. On the contrary, the combination of both the Trichoderma spp. was found to be more effective in reducing stem rot by 57.2% and increasing the survival of plants by 87.5% when compared to the individual Trichoderma applications. Further, the exogenous supplementation of Trichoderma activated antioxidative machineries, such as peroxidase, polyphenol oxidase, superoxide dismutase, catalase, and ascorbic acid in the plant. Besides, hydrogen peroxide and superoxide-free radical accumulation were also found to be reduced when T. erinaceum and T. viride were used either individually or in combination under the pathogen-challenged condition. Additionally, the photopigments in the bioprimed plants were markedly increased. Moreover, the combined inoculation of the two isolates yielded the highest records of growth parameters (root weight, shoot length, and leaf weight) compared with individual inoculation. Therefore, based on the above results, it was concluded that the combination of T. erinaceum and T. viride can be effectively used as an alternative to control white mold and stem rot caused by S. sclerotiorum.

Cyclophilin BcCyp2 Regulates Infection-Related Development to Facilitate Virulence of the Gray Mold Fungus Botrytis cinerea.

Cyclophilin (Cyp) and Ca2+/calcineurin proteins are cellular components related to fungal morphogenesis and virulence; however, their roles in mediating the pathogenesis of Botrytis cinerea, the causative agent of gray mold on over 1000 plant species, remain largely unexplored. Here, we show that disruption of cyclophilin gene BcCYP2 did not impair the pathogen mycelial growth, osmotic and oxidative stress adaptation as well as cell wall integrity, but delayed conidial germination and germling development, altered conidial and sclerotial morphology, reduced infection cushion (IC) formation, sclerotial production and virulence. Exogenous cyclic adenosine monophosphate (cAMP) rescued the deficiency of IC formation of the ∆Bccyp2 mutants, and exogenous cyclosporine A (CsA), an inhibitor targeting cyclophilins, altered hyphal morphology and prevented host-cell penetration in the BcCYP2 harboring strains. Moreover, calcineurin-dependent (CND) genes are differentially expressed in strains losing BcCYP2 in the presence of CsA, suggesting that BcCyp2 functions in the upstream of cAMP- and Ca2+/calcineurin-dependent signaling pathways. Interestingly, during IC formation, expression of BcCYP2 is downregulated in a mutant losing BcJAR1, a gene encoding histone 3 lysine 4 (H3K4) demethylase that regulates fungal development and pathogenesis, in B. cinerea, implying that BcCyp2 functions under the control of BcJar1. Collectively, our findings provide new insights into cyclophilins mediating the pathogenesis of B. cinerea and potential targets for drug intervention for fungal diseases.

Sustainable effect of a symbiotic nitrogen-fixing bacterium Sinorhizobium meliloti on nodulation and photosynthetic traits of four leguminous plants under low moisture stress environment.

Sustainable effect of a nitrogen-fixing bacterium Sinorhizobium meliloti on nodulation and photosynthetic traits (phenomenological fluxes) in four leguminous plants species under low moisture stress (20-25% soil moisture content) environment was studied. Sinorhizobium meliloti was isolated from fenugreek (Trigonella foenum-graecum) root nodules, and later, it was cultured and purified. Nodulation and photosynthetic ability in the presence of S. meliloti were tested in four leguminous plant species, that is, kidney bean (cv. lobia-2000), black bean (cv. NM-97), mung bean (cv. NM-2006) and chickpea (cv. Pb-2008). Plants of each species were grown in sterilized soil that was previously treated with 25 ml suspension containing S. meliloti at 41 × 106  CFU ml-1  kg-1 pot. One-month-old plants were subjected to low soil moisture stress conditions for 15 days, and soil moisture contents were maintained to 20-25% throughout the experimental period. The ability to fix nitrogen, nodule formation, and their subsequent effect on phenomenological fluxes in low moisture treated legumes were studied.

Leuconostoc falkenbergense sp. nov., isolated from a lactic culture, fermentating string beans and traditional yogurt.

In the present study, the taxonomic positions of five strains (C, 17-2, LMG 10779T, LMG 18969 and LMG 11483) of Leuconostoc pseudomesenteroides were re-evaluated by a polyphasic approach, including the analyses of 16S rRNA, pheS and rpoA gene sequences, cellular fatty acids, average nucleotide and amino acid identities (ANI and AAI), digital DNA-DNA hybridization (dDDH), and phenotypic features. Based on rpoA sequence analysis, the five strains and L. pseudomesenteroides LMG 11482T were divided into two groups: strains C, LMG 10779T and LMG 18969; strains 17-2, LMG 11483 and LMG 11482T. Each of the two groups had almost identical rpoA sequences. The rpoA sequence similarity between strain LMG 10779T and L. pseudomesenteroides LMG 11482T was 95.6 %. Strains LMG 11483 and 17-2 had 98.1 and 97.2 % ANI values, 83.5 and 73.2 % dDDH values, and a 97.0 % AAI value with L. pseudomesenteroides LMG 11482T, greater than the threshold for species demarcation, indicating that strains LMG 11483 and 17-2 belong to L. pseudomesenteroides. Strains LMG 18969 and C shared 97.1 and 98.2 % ANI values, 73.4 and 83.2 % dDDH values, and 96.9 and 96.6 % AAI values with strain LMG 10779T, greater than the threshold for species demarcation, indicating that strains LMG 10779T, LMG 18969 and C represent the same species. The ANI, dDDH and AAI values between strain LMG 10779T and the type strains of phylogenetically related species were 75.2-92.5, 20.0-48.2 and 75.3-93.9 %, respectively, below the thresholds for species demarcation, indicating that strain LMG 10779T represents a novel species within the genus Leuconostoc. On the basis of the results presented here, (i) strains 17-2 and LMG 11483 belong to L. pseudomesenteroides, and (ii) strains LMG 10779T, LMG 18969 and C are considered to represent a novel species within the genus Leuconostoc, for which the name Leuconostoc falkenbergense sp. nov. is proposed with the type strain LMG 10779T (=CCUG 27119T).

Paraburkholderia atlantica sp. nov. and Paraburkholderia franconis sp. nov., two new nitrogen-fixing nodulating species isolated from Atlantic forest soils in Brazil.

A polyphasic study was conducted with 11 strains trapped by Mimosa pudica and Phaseolus vulgaris grown in soils of the Brazilian Atlantic Forest. In the phylogenetic analysis of the 16S rRNA gene, one clade of strains (Psp1) showed higher similarity with Paraburkholderia piptadeniae STM7183T (99.6%), whereas the second (Psp6) was closely related to Paraburkholderia tuberum STM678T (99%). An MLSA (multilocus sequence analysis) with four (recA, gyrB, trpB and gltB) housekeeping genes placed both Psp1 and Psp6 strains in new clades, and BOX-PCR profiles indicated high intraspecific genetic diversity within each clade. Values of digital DNA-DNA hybridization (dDDH) and average nucleotide identity (ANI) of the whole genome sequences were of 56.9 and 94.4% between the Psp1 strain CNPSo 3157T and P. piptadeniae; and of 49.7% and 92.7% between the Psp6 strain CNPSo 3155T and P. tuberum, below the threshold for species delimitation. In the nodC analysis, Psp1 strains clustered together with P. piptadeniae, while Psp6 did not group with any symbiotic Paraburkholderia. Other phenotypic, genotypic and symbiotic properties were evaluated. The polyphasic analysis supports that the strains represent two novel species, for which the names Paraburkholderia franconis sp. nov. with type strain CNPSo 3157T (= ABIP 241, = LMG 31644) and Paraburkholderia atlantica sp. nov. with type strain CNPSo 3155T (= ABIP 236, = LMG 31643) are proposed.

Multiple Effect of Different Plant Growth Promoting Microorganisms on Beans (Phaseolus vulgaris L. ) Crop

Abstract We evaluated the effect of combined Rhizobium tropici, Trichoderma asperellum and plant growth-promoting rhizobacteria (PGPR) in beans crop. The hypothesis that strains of T. asperullum, R. tropici and PGPR combined could improve growth, biomass accumulation and beans yield was tested under greenhouse and field conditions. The treatments consisted of control, mineral nitrogen application and inoculation, isolated and associated with the following microorganisms: Rhizobium tropici, Bacillus subtilis, Trichoderma asperellum and Burkholderia sp. 10N6. Results were evaluated by shoot dry weight (SDW) and root dry weight (RDW), number of nodules and yield components. In greenhouse environment all the microorganisms behaved similarly, and the treatments inoculated with Burkholderia sp. 10N6 (IBu) and R. tropici (IR) stood out regarding the production components. In field conditions the treatments IR and IRTBa presented the highest values of SDW and RDW. Our results suggest that inoculation with R. tropici, T. asperellum and PGPR may promote beans growth and bring benefits to shoot and root accumulation, increase the number of nodules as well as improve yield components, contributing to a sustainable agriculture.

Structure of ribosome-bound azole-modified peptide phazolicin rationalizes its species-specific mode of bacterial translation inhibition.

Ribosome-synthesized post-translationally modified peptides (RiPPs) represent a rapidly expanding class of natural products with various biological activities. Linear azol(in)e-containing peptides (LAPs) comprise a subclass of RiPPs that display outstanding diversity of mechanisms of action while sharing common structural features. Here, we report the discovery of a new LAP biosynthetic gene cluster in the genome of Rhizobium Pop5, which encodes the precursor peptide and modification machinery of phazolicin (PHZ) - an extensively modified peptide exhibiting narrow-spectrum antibacterial activity against some symbiotic bacteria of leguminous plants. The cryo-EM structure of the Escherichia coli 70S-PHZ complex reveals that the drug interacts with the 23S rRNA and uL4/uL22 proteins and obstructs ribosomal exit tunnel in a way that is distinct from other compounds. We show that the uL4 loop sequence determines the species-specificity of antibiotic action. PHZ expands the known diversity of LAPs and may be used in the future as biocontrol agent for agricultural needs.

Rhizobium chutanense sp. nov., isolated from root nodules of Phaseolus vulgaris in China.

Two Gram-stain-negative, rod-shaped bacterial strains (C5T and C16), isolated from root nodules of Phaseolus vulgaris L. in Jiangxi Province, PR China, were characterized by using a polyphasic taxonomical approach. The phylogenetic analysis of the 16S rRNA gene and three concatenated housekeeping genes (recA-glnII-atpD) revealed that C5T and C16 were members of the genus Rhizobium, yet were distinct from known species. The case for strain C5T representing a novel species was supported by genomic results. Pairwise digital DNA-DNA hybridization and average nucleotide identity values were much lower than the proposed and generally accepted species boundaries. The genome-based phylogenetic tree reconstructed by using the up-to-date bacterial core gene set consisting of 92 genes showed that the strains formed a monophyletic branch, further supporting this result. The symbiotic genes of nodC and nifH were identified in both strains; each could nodulate Phaseolus vulgaris and Glycine max but not Leucaena leucocephala, Pisum sativum or Medicago sativa plants. Major cellular fatty acids of C5T were summed feature 8 (C18 : 1 ω7c/C18 : 1 ω6c; 58.8 %), C18 : 1 ω7c 11-methyl (14.2 %) and C18 : 0 (8.1 %). The DNA G+C content of C5T was 61.4 mol%. Based on these genomic, chemotaxonomic and phenotypic characteristics, we propose a novel species: Rhizobium chutanense sp. nov. The type strain is C5T (=CCTCC AB 2018143T=LMG 30777T).

Mesorhizobium atlanticum sp. nov., a new nitrogen-fixing species from soils of the Brazilian Atlantic Forest biome.

Biological nitrogen fixation performed by diazotrophic bacteria is a vital process for agricultural and environmental sustainability. In recent years, bacterial classification has been based on genomic data, accelerating our understanding about the diversity, and resulting in the description of several new species. In this study, four strains (CNPSo 3140T, CNPSo 3235, CNPSo 3236 and CNPSo 3237) trapped by Phaseolus vulgaris and Mimosa pudica from soil samples of the Brazilian Atlantic Forest biome (Mata Atlântica) were submitted to polyphasic analysis to investigate their proper classification within the genus Mesorhizobium. The 16S rRNA gene phylogram showed that the strains present sequences identical to those of Mesorhizobium acaciaeand Mesorhizobium plurifarium, not allowing a clear taxonomic classification; however, when using multilocus sequence analysis methodology, the strains were grouped into a well-supported distinct clade, with <94.5 % nucleotide identity with the other species of the genus. The average nucleotide identity of CNPSo 3140T genome showed values below the threshold in relation to the closest species, of 89.75 % with Mesorhizobium plurifariumand of 88.83 % with Mesorhizobium hawassense; the digital DNA-DNA hybridization values were 39 and 37.70 % with the same species, respectively. Nodulation gene (nodC) phylogeny positioned the strains in an isolated cluster, showing greater similarity to Mesorhizobiumshonense. All data obtained in this study support the description of the novel species Mesorhizobiumatlanticum sp. nov. The type strain is CNPSo 3140T (=ABIP 206T=LMG 30305T=U1602T), isolated from a nodule of Phaseolus vulgaris.

The repertoire of effector candidates in Colletotrichum lindemuthianum reveals important information about Colletotrichum genus lifestyle.

The fungus Colletotrichum lindemuthianum is the causal agent of anthracnose in the common bean (Phaseolus vulgaris), and anthracnose is one of the most devastating diseases of this plant species. However, little is known about the proteins that are essential for the fungus-plant interactions. Knowledge of the fungus' arsenal of effector proteins is of great importance for understanding this pathosystem. In this work, we analyzed for the first time the arsenal of Colletotrichum lindemuthianum effector candidates (ClECs) and compared them with effector proteins from other species of the genus Colletotrichum, providing a valuable resource for studying the infection mechanisms of these pathogens in their hosts. Isolates of two physiological races (83.501 and 89 A2 2-3) of C. lindemuthianum were used to predict 353 and 349 ClECs, respectively. Of these ClECs, 63% were found to be rich in cysteine, have repetitive sequences of amino acids, and/or possess nuclear localization sequences. Several conserved domains were found between the ClECs. We also applied the effector prediction to nine species in the genus Colletotrichum, and the results ranged from 247 predicted effectors in Colletotrichum graminicola to 446 in Colletotrichum orbiculare. Twelve conserved domains were predicted in the effector candidates of all analyzed species of Colletotrichum. An expression analysis of the eight genes encoding the effector candidates in C. lindemuthianum revealed their induction during the biotrophic phase of the fungus on the bean.

Cysteine-Rich Receptor-Like Kinase Gene Family Identification in the Phaseolus Genome and Comparative Analysis of Their Expression Profiles Specific to Mycorrhizal and Rhizobial Symbiosis.

Receptor-like kinases (RLKs) are conserved upstream signaling molecules that regulate several biological processes, including plant development and stress adaptation. Cysteine (C)-rich receptor-like kinases (CRKs) are an important class of RLK that play vital roles in disease resistance and cell death in plants. Genome-wide analyses of CRK genes have been carried out in Arabidopsis and rice, while functional characterization of some CRKs has been carried out in wheat and tomato in addition to Arabidopsis. A comprehensive analysis of the CRK gene family in leguminous crops has not yet been conducted, and our understanding of their roles in symbiosis is rather limited. Here, we report the comprehensive analysis of the PhaseolusCRK gene family, including identification, sequence similarity, phylogeny, chromosomal localization, gene structures, transcript expression profiles, and in silico promoter analysis. Forty-six CRK homologs were identified and phylogenetically clustered into five groups. Expression analysis suggests that PvCRK genes are differentially expressed in both vegetative and reproductive tissues. Further, transcriptomic analysis revealed that shared and unique CRK genes were upregulated during arbuscular mycorrhizal and rhizobial symbiosis. Overall, the systematic analysis of the PvCRK gene family provides valuable information for further studies on the biological roles of CRKs in various Phaseolus tissues during diverse biological processes, including Phaseolus-mycorrhiza/rhizobia symbiosis.

Use of plant growth promoting Rhizobacteria (PGPR) and mycorrhizae to improve the growth and nutrient utilization of common bean in a soil infected with white rot fungi.

Extensive use of fertilizers and pesticides led to dangerous ecological effects and therefore the biological approaches have been widely recommended to prevent further deterioration for the environment. The current study was conducted to explore the potentiality of using single or combined inoculations by mycorrhizae, Bacillus subtilis and Pseudomonas fluorescence for controlling the infection of common bean plants with Sclerotium rolfsii on one hand and as bio-fertilizers for improving plants nutritional status on the other hand. The soil of study was mildly infected with S. rolfsii and contained high total-P content. Thus, minimal P inputs were added to the inoculated soil in the form of rock phosphate. Activities of plant defense enzymes i.e. chitinase, peroxidase and polyphenol oxidase were determined under the greenhouse conditions and the results obtained herein indicated that activities of such enzymes increased significantly owing to bio-agent inoculations. In this concern, combined treatments resulted in further significant increases over the single ones. A field study was then conducted for two successive years and the results reveal that single inoculations increased straw and green pod yields as well as the uptake of P and Fe by plants as compared with the non-inoculated treatment. Combined inoculants recorded further significant increases in these parameters even when compared with the fungicide treated plants. Generally, straw and pod yields obtained from the second growing season were significantly higher than those attained in the first growing one. Our study confirms the success of the used bio-treatments in minimizing soil pollution through fertilizer and/or pesticide inputs.

Essential oils of Tahiti lemon and cinnamon bark in control of storage fungi and the physiological and sanitary quality of beans / Óleos essenciais de limão Taiti e canela em casca no controle de fungos de armazenamento e na qualidade fisiológica e sanitária de feijão

With the objective to evaluate the efficiency of essential oils of Citrus latifolia (Tahiti lemon) and Cinnamomum zeylanicum (cinnamon bark) in the control of plant pathogens Penicillium sp. and Aspergillus sp. and the quality of the bean seeds, two experiments were conducted. In the first one, the effect of essential oils of C. latifolia and C. zeylanicum was evaluated in vitro development of the fungi Penicillium sp. and Aspergillus sp. and, in the second one, the influence of essential oils on the physiological and sanitary quality of bean seeds. The variables mycelial growth, conidial germination and sporulation of Penicillium sp. and Aspergillus sp. were measured in the first experiment, while the seed germination test, first count of germination, germination speed index (GSI) and sanity test of bean seeds were measured in the second. The essential oil (EO) of C. zeylanicum was more efficient than C. latifolia in the control of Aspergillus sp. and Penicillium sp., but decreased the physiological quality of the beans seeds. The fungal diversity identified in the seed health test was composed by fungi of the genera Aspergillus, Penicillium, Cladosporium, Fusarium, Chaetomium and Rhizopus. The results indicate the potential of the use of these EOs in the seeds treatment.(AU)

Com o objetivo de avaliar a eficiência dos óleos essenciais de Citrus latifolia (limão taiti) e Cinnamomum zeylanicum (canela em casca) no controle dos fitopatógenos Penicillium sp. e Aspergillus sp. e na qualidade das sementes de feijão, foram conduzidos dois experimentos. No primeiro, avaliou-se o efeito dos óleos essenciais de C. latifolia e C. zeylanicum no desenvolvimento in vitro dos fungos Penicillium sp. e Aspergillus sp. e, no segundo, a influência dos óleos essenciais sobre a qualidade fisiológica e sanitária das sementes de feijão. As variáveis crescimento micelial, germinação de conídios e esporulação de Penicillium sp. e Aspergillus sp. foram aferidas no primeiro experimento, enquanto o teste de germinação de sementes, primeira contagem de germinação, índice de velocidade de germinação (IVG) e teste de sanidade de sementes de feijão foram aferidas no segundo. O óleo essencial (OE) de C. zeylanicum foi mais eficiente que C. latifolia no controle dos fungos Aspergillus sp. e Penicillium sp., mas diminuiu a qualidade fisiológica das sementes de feijão. A diversidade fúngica identificada no teste de sanidade de sementes foi composta por fungos dos gêneros Aspergillus, Penicillium, Cladosporium, Fusarium, Chaetomium e Rhizopus. Os resultados indicam o potencial do uso desses óleos essenciais no tratamento de sementes.(AU)

Biochar and Trichoderma harzianum for the Control of Macrophomina phaseolina

Abstract This study is based on the importance of biological control methods and the lack of information on the effect of biochar (BCH) from sewage sludge associated or not with Trichoderma harzianum on the control of Macrophomina phaseolina in the bean crop (Phaseolus vulgaris, cv. BRS Estilo). Biochar from sewage sludge, pyrolyzed at 500 ºC and used in low concentration (0.5%), has a direct effect on the in vitro control of M. phaseolina. However, higher BCH concentrations stimulated the growth of the pathogen. In culture medium with or without BCH, T. harzianum (strain 1306) inhibited the mycelial growth of M. phaseolina. The addition of BCH + T. harzianum reduced the deleterious effects caused by M. phaseolina on bean plants. This study demonstrated that joint application of BCH from sewage sludge + T. harzianum considerably increased the fresh and dry mass of bean plants, inoculated or not with M. phaseolina.

Biochemical responses of common bean to white mold potentiated by phosphites.

Considering that the mechanisms for phosphite-afforded disease control remain elusive, this study investigated whether zinc (Zn) and copper (Cu) phosphites could possible potentiate common bean resistance to white mold, caused by Sclerotinia sclerotiorum, through the stimulation of biochemical defence responses. Lesion area and disease severity were decreased by phosphites spray, but Zn phosphite outcompeted Cu phosphite. Histopathological observations revealed fewer fungal hyphae and less collapse of the mesophyll cells in the Zn and Cu phosphite-sprayed plants compared to water-sprayed ones. The S. sclerotiorum-triggered accumulation of reactive oxygen species, oxalic acid (a fungal secreted toxin) and malondialdehyde (an indicator of cellular damage) were constrained as a result of Zn and Cu phosphites spray. Activities of antioxidant enzymes (superoxide dismutase, peroxidase, ascorbate peroxidase and glutathione-S-transferase at 12 h after inoculation (hai) and catalase at 60 and 84 hai) were higher for Zn and Cu phosphites-sprayed plants than for water-sprayed ones. Activities of defence-related enzymes chitinase (CHI) at 12 hai, ß-1,3-glucanase (GLU) and polyphenoloxidase (PPO) were higher at 12-84 hai for Zn, and Cu phosphites sprayed plants, phenylalanine ammonia-lyase at 36-84 hai for the Zn phosphite sprayed ones, CHI at 12-36 hai, GLU at 12-60 hai, PPO at 36 hai and PAL and lipoxygenase at 12 hai for the Cu phosphite sprayed ones upon inoculation with S. sclerotiorum relative to their water-sprayed counterparts. Concentrations of total soluble phenols and lignin-thioglycolic acid derivatives were not affected by Cu phosphite spray on infected plants but were higher and lower, respectively, for Zn phosphite sprayed plants at 60 hai compared to water-sprayed ones. Taken together, the findings from the present study shed light on the biochemical defence mechanisms involved in the Zn and Cu phosphites-mediated suppression of white mold in common bean.

BcXYG1, a Secreted Xyloglucanase from Botrytis cinerea, Triggers Both Cell Death and Plant Immune Responses.

In search of Botrytis cinerea cell death-inducing proteins, we found a xyloglucanase (BcXYG1) that induced strong necrosis and a resistance response in dicot plants. Expression of the BcXYG1 gene was strongly induced during the first 12 h post inoculation, and analysis of disease dynamics using PathTrack showed that a B. cinerea strain overexpressing BcXYG1 produced early local necrosis, supporting a role of BcXYG1 as an early cell death-inducing factor. The xyloglucanase activity of BcXYG1 was not necessary for the induction of necrosis and plant resistance, as a mutant of BcXYG1 lacking the xyloglucanase enzymatic activity retained both functions. Residues in two exposed loops on the surface of BcXYG1 were found to be necessary for the induction of cell death but not to induce plant resistance. Further analyses showed that BcXYG1 is apoplastic and possibly interacts with the proteins of the plant cell membrane and also that the BcXYG1 cell death-promoting signal is mediated by the leucine-rich repeat receptor-like kinases BAK1 and SOBIR1. Our findings support the role of cell death-inducing proteins in establishing the infection of necrotrophic pathogens and highlight the recognition of fungal apoplastic proteins by the plant immune system as an important mechanism of resistance against this class of pathogens.

Bradyrhizobium sacchari sp. nov., a legume nodulating bacterium isolated from sugarcane roots.

Members of the genus Bradyrhizobium are well-known as nitrogen-fixing microsymbionts of a wide variety of leguminous species, but they have also been found in different environments, notably as endophytes in non-legumes such as sugarcane. This study presents a detailed polyphasic characterization of four Bradyrhizobium strains (type strain BR 10280T), previously isolated from roots of sugarcane in Brazil. 16S rRNA sequence analysis, multilocus sequence analysis (MLSA) and analysis of the 16S-23S rRNA internal transcribed spacer showed that these strains form a novel clade close to, but different from B. huanghuaihaiense strain CCBAU 23303T. Average nucleotide identity (ANI) analyses confirmed that BR 10280T represents a novel species. Phylogenetic analysis based on nodC gene sequences also placed the strains close to CCBAU 23303T, but different from this latter strain, the sugarcane strains did not nodulate soybean, although they effectively nodulated Vigna unguiculata, Cajanus cajan and Macroptilium atropurpureum. Physiological traits are in agreement with the placement of the strains in the genus Bradyrhizobium as a novel species for which the name Bradyrhizobium sacchari sp. nov. is proposed.