Harpin_(xoo) and Its Functional Domains Activate Pathogen-inducible Plant Promoters in Arabidopsis

Harpins are bacterial proteins that can enhance plant growth and defense against pathogens and insects. To elaborate whether harpins perform the diverse functions in coordination with the activation of specific promoters that contain particular elements, we cloned pathogen-inducible plant promoters PPP1, PPP2, and PPP3 from tobacco and investigated their responses to harpinxoo or its truncated fragments DEG, DIR, and DPR (domains for enhancing plant growth, insect resistance and pathogen resistance). PPP1 contains an internal repeat composed of two tandem 111bp fragments; 111bp in the repeat was deleted in PPP2. PPP3 contains a bacteria-inducible element; PPP1 and PPP2 additionally contain TAC-1 and Eli boxes inducible correspondingly by salicylic acid (SA) and elicitors. Function of cloned PPPs was confirmed based on their activation in transgenic Arabidopsis plants by Ralstonia solanacearum (Ralston) or SA. Harpinxoo, DEG, DIR, or DPR activated PPP1 and PPP2 but not PPP3, consistent with the presence of Eli boxes in promoters. PPP1 was ca. 3-fold more active than PPP2, suggesting that the internal repeat affects levels of the promoter activation.

Utilizing the γ-Irradiated Sodium Alginate as a Plant Growth Promoter for Enhancing the Growth,Physiological Activities,and Alkaloids Production in Catharanthus roseus L.

Sodium alginate is a polysaccharide that is largely obtained from the brown algae （Sargassum sp.）.It has been used as a wonderful growth promoting substance in its depolymerized form for various plants.The aim of this study was to find out the effects of various concentrations of γ-irradiated sodium alginate （ISA）,viz.,deionized water （control,T0）,20 （T1）,40 （T2）,60 （T3）,80 （T4）,and 100 ppm （T5） on the agricultural performance of Catharanthus roseus L.（Rosea） in terms of growth attributes,photosynthesis,physiological activities,and alkaloid production.The present work revealed that ISA applied as leaf-sprays at concentrations from 20 to 100 ppm might improve growth,photosynthesis,physiological activities,and alkaloid production in C.roseus L.significantly.Of the various ISA concentrations,80 ppm proved to be the best one compared to other concentrations applied.

Endophytic Bacteria Promote the Growth of Suaeda glauca in Saline-Alkali Stress:Regulation of Osmotic Pressure and Antioxidative Defense System

Endophytic bacteria are promising bacterial fertilizers to improve plant growth under adverse environment.For ecological remediation of coastal wetlands,it was necessary to investigate the effect and interaction of endophytes on halophytes under saline-alkali stress.In this study,an endophytic bacterium strain HK1 isolated from halophytes was selected to infect Suaeda glauca under pH(7 and 8)and salinity gradient(150,300 and 450mmolL^(-1)).Strain HK1 was identified as Pantoea ananatis and it had ability to fix nitrogen,dissolve inorganic phosphorus and produce indole-3-aceticacid(IAA).The results showed that strain HK1 could promote the growth of S.glauca seedings when the salinity was less than 300mmolL^(-1),in view of longer shoot length and heavier fresh weight.The infected plants could produce more proline to decrease the permeability of cells,which content increased by 26.2%–61.1%compared to the non-infected group.Moreover,the oxidative stress of infected plants was relieved with the malondialdehyde(MDA)content decreased by 16.8%–32.9%,and the peroxidase(POD)activity and catalase(CAT)activity increased by 100%–500%and 6.2%–71.4%,respectively.Statistical analysis revealed that increasing proline content and enhancing CAT and POD activities were the main pathways to alleviate saline-alkali stress by strain HK1 infection,and the latter might be more important.This study illustrated that endophytic bacteria could promote the growth of halophytes by regulation of osmotic substances and strengthening antioxidant activities.This finding would be helpful for the bioremediation of coastal soil.

Research Progress of Mechanism of Action of Plant Growth Promoting Rhizobacteria

The paper first introduces the definition and classification of plant growth promoting rhizobacteria （PGPR）, then reviews the research achievements on the mechanism of action of plant growth promoting rhizobacteria, including growth pro-moting mechanism and bio-control mechanism, subsequently lists the use of excel-lent plant growth promoting rhizobacteria strains in recent years, especial y Pseu-domonas and Bacil us strains, and final y discusses problems existing in this area and points out issues requiring further exploration, including PGPR screening meth-ods, preservation methods, mechanism of action, in order to commercialize PGPR as soon as possible and practical y realize its application to production.

Rhizosphere Bacteria for Biocontrol of Bacterial Blight and Growth Promotion of Rice

Several bacterial strains were isolated from different rhizospheres. Among these, strain PDY7 exhibited strong antibacterial activity against the rice bacterial blight （BB） pathogen Xanthomonas oryzae pv. oryzae （Xoo） by the laboratory dual plate assays. The antibacterial property of the strain PDY7 was further investigated for the production of 2,4-diacetylphloroglucinol （DAPG）, which amplified a characteristic of 629-bp DNA fragment by PCR-based screening method using phlD primers. The application of phlD positive strains was carefully evaluated for disease control and growth promotion of rice plants under field conditions. The selected strain PDY7 suppressed the rice BB by 58.83% and 51.88% under glass house and field conditions, respectively. In addition, the strain PDY7 showed significant two-fold increase in root length （18.08 cm）, shoot length （29.81 cm）, and grain yield （96.07 g）. Strain PDY7 promoted the growth of rice plants by production of indole-3-acetic acid （IAA）, which was determined by high performance liquid chromatography （HPLC） analysis. Our findings suggest that PDY7 belongs to the P. fluorescens group and can serve as potential biocontrol of BB as well as biofertilizer agent for growth promotion of rice.

Transcriptomic insights into growth promotion effect of Trichoderma afroharzianum TM2-4 microbial agent on tomato plants

Plant growth promoting fungi are receiving increased attention as valuable beneficial microorganisms in crop cultivation due to their capacity to produce bioactive substances,promote plant growth and enhance immune defense functions.In this study,a novel Trichoderma isolate,designated as TM2-4,was screened from healthy tomato rhizosphere soil and identified as Trichoderma afroharzianum.Culture filtrate of the isolate TM2-4 displayed obvious bioactive substance production and an evident effect in promoting tomato seed germination,with hypocotyl length,radical length and vigor index increased by 28.7,19.4 and 62.1%,respectively,after a 100-fold dilution treatment.To assess the promotion effect and related mechanism of isolate TM2-4,the plant biological indexes and gene expression profiles of tomato plants treated with or without T.afroharzianum TM2-4 microbial agent were investigated by greenhouse pot experiment and RNA sequencing.The results demonstrated that T.afroharzianum TM2-4 significantly promoted tomato plant growth in terms of plant height,dry weight,number of leaves per plant and root activity,through efficient colonization in the rhizosphere and root system of the plants.Transcriptome analyses identified a total of 984 differentially expressed genes in T.afroharzianum microbial agent inoculated tomato roots,which were mainly engaged in the biological process of phytohormone homeostasis,antioxidant activity,as well as metabolic pathways including phenylpropanoid biosynthesis and glutathione metabolism.These findings provide useful information for understanding the mechanism of isolate TM2-4 for tomato plant growth promotion,which would facilitate further development of T.afroharzianum TM2-4 microbial agent for use in vegetable crop production.

Mycorrhizoremediation—an enhanced form of phytoremediation

Study of plant roots and the diversity of soil micro biota, such as bacteria, fungi and microfauna associated with them, is important for understanding the ecological complexities between diverse plants, microbes, soil and climates and their role in phytoremediation of contaminated soils. The arbuscular mycorrhizal fungi (AMF) are universal and ubiquitous rhizosphere mi-croflora forming symbiosis with plant roots and acting as biofertilizers, bioprotactants, and biodegraders. In addition to AMF, soils also contain various antagonistic and beneficial bacteria such as root pathogens, plant growth promoting rhizobacteria including free-living and symbiotic N-fixers, and mycorrhiza helping bacteria. Their potential role in phytoremediation of heavy metal (HM) contaminated soils and water is becoming evident although there is need to completely understand the ecological complexities of the plant-microbe-soil interactions and their better exploitation as consortia in remediation strategies employed for contaminated soils. These multitrophic root microbial associations deserve multi-disciplinary investigations using molecular, biochemical, and physiological techniques. Ecosystem restoration of heavy metal contaminated soils practices need to incorporate microbial bio-technology research and development. This review highlights the ecological complexity and diversity of plant-microbe-soil combinations, particularly AM and provides an overview on the recent developments in this area. It also discusses the role AMF play in phytorestoration of HM contaminated soils, i.e. mycorrhizoremediation.

Increased ammonification,nitrogenase,soil respiration and microbial biomass N in the rhizosphere of rice plants inoculated with rhizobacteria

Azospirillum brasilense and Pseudomonas fluorescens are well-known plant growth promoting rhizobacteria.However,the effects of A.brasilense and P.fluorescens on the N cycles in the paddy field and rice plant growth are little known.This study investigated whether and how A.brasilense and P.fluorescens contribute to the N transformations and N supply capacities in the rhizosphere,and clarified the effects of A.brasilense and P.fluorescens on the N application rate in rice cultivation.Inoculations with A.brasilense and P.fluorescens coupled with N application rate trials were conducted in the paddy field in 2016 and 2017.The inoculations of rice seedlings included four treatments:sterile saline solution(M_(0)),A.brasilense(M_(b)),P.fluorescens(M_(p)),and co-inoculation with a mixture of A.brasilense and P.fluorescens(M_(bp)).The N application rate included four levels:0 kg N ha^(–1)(N_(0)),90 kg N ha^(–1)(N_(90)),180 kg N ha^(–1)(N_(180)),and 270 kg N ha^(–1)(N_(270)).The results indicated that the M_(bp) and M_(p) treatments significantly enhanced the ammonification activities in the rhizosphere compared with the M_(0) treatment,especially for higher N applications,while the Mbp and M_(b) treatments greatly enhanced the nitrogenase activities in the rhizosphere compared with the M_(0) treatments,especially for lower N applications.Azospirillum brasilense and P.fluorescens did not participate in the nitrification processes or the denitrification processes in the soil.The soil respiration rate and microbial biomass N were greatly affected by the interactions between the rhizobacteria inoculations and the N fertilizer applications.In the M_(bp) treatment,N supply capacities and rice grain yields showed no significant differences among the N_(90),N_(180),and N_(270) applications.The N application rate in the study region can be reduced to 90 kg N ha^(–1) for rice seedlings co-inoculated with a mixture of A.brasilense and P.fluorescens.

The rhizosphere microbial complex in plant health:A review of interaction dynamics

Climate change,urbanization,and population increase limit food availability.To sustain human existence,there is the need to increase food and agricultural production to mitigate the impact of these factors.Scientists have been working for years on ways to increase food production.From plant breeding techniques to soil science,scientists have made tremendous progress.The rhizobiome has been proven to be important to crop production,and the impact of the rhizobiome on plant health cannot be overemphasized.Being rich in diverse complex microbial interactions,the rhizosphere has become a major force in recent plant growth promotion studies.The upsurge in next-generation sequencing applications with the various“omics”technologies is helping to unearth information relating to rhizosphere impact on plant growth.Explaining the complex interactions between and across microbial species present in the rhizosphere is important to further enhance our understanding of their mechanistic and mutualistic functions.Knowledge from this can be used in rhizosphere biome engineering for improved plant growth and yield in the face of the various biotic and abiotic challenges.

Potential of Biostimulants Based on PGPR Rhizobacteria Native to Benin’s Soils on the Growth and Yield of Maize (<i>Zea mays </i>L.) under Greenhouse Conditions

The application of biostimulants in agriculture represents an environmentally friendly alternative while increasing agricultural production. The aims of the study were to develop solid biostimulants based on five rhizobacteria native to Benin’s soils and to evaluate their efficacy on the growth and biomass yield of maize under greenhouse conditions on ferrallitic and ferruginous soils. Clay and peat were used as a conservation binder for the preparation of the biostimulants. These binders were used alone or combined in the different formulations with maize flour and sucrose. 10 g of biostimulants were applied at sowing in pots containing five kilograms of sterilised soil. The experimental design was a completely randomised block of 24 treatments with three replicates. The results obtained showed significant improvements (P < 0.001) in height (49.49%), stem diameter (32.7%), leaf area (66.10%), above-ground biomass (97.12%) and below-ground biomass (53.98%) on ferrallitic soil with the application of the clay + Pseudomonas putida biostimulant compared to the control. On the other hand, the use of the peat biostimulant + Pseudomonas syringae was more beneficial for plant growth on ferruginous soil. The height, stem diameter, leaf area, above-ground biomass and below-ground biomass of the plants under the influence of this biostimulant were improved by 83.06%, 44.57%, 102.94%, 86.84% and 42.68%, respectively, compared to the control. Therefore, these results confirm that Rhizobacteria express their potential through biostimulants formulated on maize. The formulated biostimulants can later be used by producers to improve crop productivity for sustainable agriculture.

Phenotypic, Stress Tolerance and Plant Growth Promoting Characteristics of Rhizobial Isolates from Selected Wild Legumes of Semiarid Region, Tirupati, India

Rhizobia are vital for nitrogen input, fertility of soil and legume plant growth. Knowledge on rhizobial diversity from arid and semiarid areas is important for dry land agriculture in the context of climatic change and for economic utilization. This study provides morphological, biochemical, stress tolerance and plant growth promoting characteristics of fifteen rhizobial isolates from the nodules of same number of wild legumes and one isolate from cultivated Arachis hypogea from semi-arid region, Tirupati. The bacterial isolates were confirmed as rhizobia based on colony morphology and biochemical tests. Based on the colour change of YMA-BTB medium, eight isolates were identified as slow growers and six were fast growers. The isolates differed in growth pattern, colony morphology, antibiotic resistance at higher concentrations and uniformity in utilization of carbon and nitrogen sources. The isolates are tolerant to NaCl up to one percent, displayed normal growth at temperatures 28℃ - 30℃, at neutral pH and poor growth at pH 5and 9. The isolates varied in the production of EPS and IAA, positive for phosphate solubilization and siderophore formation. This functional diversity displayed by the isolates can be utilised for the legume crop production by cross inoculation.

Evaluation of Plant Growth Promoting Ability of Bacillus amyloliquefaciens Bc2 and Trichoderma harzianum TR In Vivo

Bacillus amyloliquefaciens Bc2 and Trichderma harzianum TR were used to evaluate their growth-promoting activity on cultivated strawberries, under laboratory and field conditions, and we have noticed that the percentage of achene germination is important for ones treated with TR (=97%) followed by those treated with Bc2 strain (=90%) and the control (=84%). Inoculations on field showed that on untreated soil with insecticide, TR is effective and allows the development of plants and extends the duration of flowering and fruiting. On treated soil, Bc2 clearly promotes the growth and development of strawberry seedlings and its role as plant growth promoting microorganisms has been proved.

Improvement of Sporulation Conditions of a New Strain of Bacillus amyloliquefaciens in Liquid Fermentation

The desirable active ingredients for the development of bioproducts based in Bacillus sp. for the control of soil pathogens are the spores because these structures exhibit more resistance and stability to conditions present during the fermentation, formulation, and storage processes. To improve the sporulation of a native strain of Bacillus amyloliquefaciens (Bs006) using liquid fermentation, some modifications in the concentrations of the components in a previously standardized culture media were made. Subsequently, five sporulation inducers (iron nitrate, mixture of salts, peroxide hydrogen, temperature, and initial cell concentration) were evaluated. The treatment with a mixture of salts in combination with an initial cell concentration of 1 × 108 cells/ml was selected because a final spore concentration of 1.05 × 1010 spores/ml after 66 hours with a fully substrate consumption and sporulation efficiency of 88.7% was obtained. To demonstrate the biological activity of B. amyloliquefaciens Bs006 in Cape gooseberry seedlings, a greenhouse bioassay was conducted, and statistical differences in plant growth-promoting parameters compared with previous media were not found. Additionally, the proposed modified media (coded as JM) presented a benefit-cost ratio 2.65 times higher compared with the baseline media.

Research Progress in Biological Control of Soft Rot of Amorphophallus konjac

In this paper,the main control methods of soft rot of Amorphophallus konjac are reviewed,with a focus on the current research status of using plant growth promoting rhizobacteria for biological control of soft rot of A.konjac,and future research directions are looked forward to.

Plant growth-promoting characteristics of root fungal endophytes isolated from a traditional Cordillera rice landrace

This study was conducted to isolate and characterize the plant growth-promoting potential of fungal endophytes from the roots of Diket red,a traditional rice plant from the Cordillera,Northern Luzon.Eighteen morphospecies of filamentous endophytes were isolated of which twelve isolates were successfully identified to the species level.These isolates were identified as Aspergillus versicolor,Aspergillus sp.,Chaetosphaeria sp.,Cladosporium cladosporioides,Hypocrea lixii,Microascus murinus,and Trichoderma harzianum.The identified twelve isolates were selected to screen in vitro for their plant growth-promoting characteristics,and evaluated in vivo for their beneficial effects on seedling vigor and early seedling growth.Isolate PPL14 produced the highest IAA at 55.5μg ml^(-1)and M.murinus PPL10 produced the highest amount of IAA at 3.73μg IAA mg^(-1)dry mycelia wt.The seedling vigor assay and in vivo plant growth promotion bioassay indicated overall positive effects of culture filtrate(CF)application of the endophyte isolates.Rice seeds and seedlings grown in aseptic condition and treated with endophyte CFs displayed significantly enhanced levels of germination,seedling vigor,shoot,root,and total plant growth,and biomass compared to non-treated control.Other plant growth-promoting characteristics were also studied including phosphate solubilization,siderophore production,ammonia production,and catalase activity.This study supports the potential use of fungal endophytes as bio-inoculants for plant growth promotion and enhancement of nutrient assimilation of agriculturally important crops.

Shortened snRNA promoters for efficient CRISPR/Cas-based multiplex genome editing in monocot plants

Dear Editor,CRISPR(clustered regularly interspaced short palindromic repeats)/Cas genome editing is a powerful tool for introducing specific mutations in organisms including plants.The system is composed of a nuclease such as Cas9 or Cas12a and an engineered single-guide RNA(sgRNA)incorporating a target sequence(Li et al.,2019).A Cas9/sgRNA complex recognizes its target site in the genome,resulting in a mutation at that site.

Finding Signals for Plant Promoters

The strongest signal of plant promoter is searched with the model of single motif with two types. It turns out that the dominant type is the TATA-box. The other type may be called TATA-less signal, and may be used in gene finders for promoter recognition. While the TATA signals are very close for the monocot and the dicot, their TATA-less signals are significantly different. A general and flexible multi-motif model is also proposed for promoter analysis based on dynamic programming. By extending the Gibbs sampler to the dynamic programming and introducing temperature, an efficient algorithm is developed for searching signals in plant promoters.

Toxicity of copper oxide nanoparticles on agriculturally important soil rhizobacteria Bacillus megaterium

Copper oxide nanoparticles are an integral part of modern agrochemical industries,such as pesticides and nano fertilizers.It has been extensively reviewed and reported that the interaction of nanoparticles with plant growth-promoting rhizobacteria(PGPRs)may be toxic due to their accumulation in the soil and may remain in contact for a long time which causes a negative impact on the soil microbial community.The study presented here,investigates the impact of the copper oxide nanoparticles on the isolated PGPR species Bacillius megaterium AGNTL MM7 in invitro conditions.We observed that the formation of Reactive Oxygen Species(ROS)at¼of Minimum Inhibitory Concentrations(MIC)of CuO nanoparticles resulted in a decrease in the Indole Acetic Acid(IAA)production(83.02μg/ml)as the marker of the nanoparticles mediated stress on the Bacillus megaterium AGNTL MM7.The minimum inhibitory concentration(MIC)assay was observed at 4 mg/ml.The solubility of CuO nanoparticles was 0.014%and the Nanotracking analyzer estimated the mean average size 188 nm in a liquid medium such as water.Transmission Electron Microscopic(TEM)studies observed that the concentration of 1 mg/ml nanoparticles showed a strong affinity with cell envelope and caused a significant negative impact on the microorganism,resulting in 58%inhibition in IAA production was observed in comparison to the control i.e.,197.8μg/ml.The flow cytometric analysis also significantly validated the toxicity at the same concentration.

Difficult-to-culture bacteria in the rhizosphere:The underexplored signature microbial groups

Microorganisms represent a substantial portion of the earth’s biodiversity and biomass, and the plant rhizosphere is an innate reservoir teeming with heterogeneous microbes predominated by bacterial communities. Rhizospheric microbial diversity (genetic, phenotypic, and metabolic) has been extensively studied to understand the key ecological roles played by the microbial members, including plant growth promotion. The application of 16S rRNA gene sequencing and next-generation sequencing (NGS) technologies has revolutionized the discovery of novel bacterial groups that have remained undetected by traditional cultivation-based approaches. Such technological advancements have opened new vistas in our current understanding of predominant but concealed and missed bacterial diversity referred to as difficult-to-culture bacterial lineages, especially the predominant phyla Acidobacteria, Verrucomicrobia,Planctomycetes, and Gemmatimonadetes. Regardless of their ubiquity and prevalence, little is known about their ecophysiology because of the non-availability of culturable members. More recently, there has been increased interest in understanding the cosmopolitan distribution and diversity of the difficult-to-culture bacteria, focusing on their role in driving complex plant-microbial interactions and mobilizing nutrients in soil and their potential as sources of novel bioactive metabolites. As an initial step, we review the distribution and significance of such bacterial phyla in soil, their ecophysiological roles, and their hidden plant growth promoting potential. The ability to select and deploy plant probiotic bacteria from the difficult-to-culture fraction of the bacterial community might open new avenues for improving crop health.