Whole genome sequencing of Enterobacter mori, an emerging pathogen of kiwifruit and the potential genetic adaptation to pathogenic lifestyle

Members of the Enterobacter genus are gram-negative bacteria, which are used as plant growth-promoting bacteria, and increasingly recovered from economic plants as emerging pathogens. A new Enterobacter mori strain, designated CX01, was isolated as an emerging bacterial pathogen of a recent outbreak of kiwifruit canker-like disease in China. The main symptoms associated with this syndrome are bleeding cankers on the trunk and branch, and brown leaf spots. The genome sequence of E. mori CX01 was determined as a single chromosome of 4,966,908 bp with 4640 predicted open reading frames (ORFs). To better understand the features of the genus and its potential pathogenic mechanisms, five available Enterobacter genomes were compared and a pan-genome of 4870 COGs with 3158 core COGs were revealed. An important feature of the E. mori CX01 genome is that it lacks a type III secretion system often found in pathogenic bacteria, instead it is equipped with type I, II, and VI secretory systems. Besides, the genes encoding putative virulence effectors, two-component systems, nutrient acquisition systems, proteins involved in phytohormone synthesis, which may contribute to the virulence and adaption to the host plant niches are included. The genome sequence of E. mori CX01 has high similarity with that of E. mori LMG 25,706, though the rearrangements occur throughout two genomes. Further pathogenicity assay showed that both strains can either invade kiwifruit or mulberry, indicating they may have similar host range. Comparison with a closely related isolate enabled us to understand its pathogenesis and ecology.

Electrical Signals, Plant Tolerance to Actions of Stressors, and Programmed Cell Death: Is Interaction Possible?

In environmental conditions, plants are affected by abiotic and biotic stressors which can be heterogenous. This means that the systemic plant adaptive responses on their actions require long-distance stress signals including electrical signals (ESs). ESs are based on transient changes in the activities of ion channels and H+-ATP-ase in the plasma membrane. They influence numerous physiological processes, including gene expression, phytohormone synthesis, photosynthesis, respiration, phloem mass flow, ATP content, and many others. It is considered that these changes increase plant tolerance to the action of stressors; the effect can be related to stimulation of damages of specific molecular structures. In this review, we hypothesize that programmed cell death (PCD) in plant cells can be interconnected with ESs. There are the following points supporting this hypothesis. (i) Propagation of ESs can be related to ROS waves; these waves are a probable mechanism of PCD initiation. (ii) ESs induce the inactivation of photosynthetic dark reactions and activation of respiration. Both responses can also produce ROS and, probably, induce PCD. (iii) ESs stimulate the synthesis of stress phytohormones (e.g., jasmonic acid, salicylic acid, and ethylene) which are known to contribute to the induction of PCD. (iv) Generation of ESs accompanies K+ efflux from the cytoplasm that is also a mechanism of induction of PCD. Our review argues for the possibility of PCD induction by electrical signals and shows some directions of future investigations in the field.

Bacillus circulans GN03 Alters the Microbiota, Promotes Cotton Seedling Growth and Disease Resistance, and Increases the Expression of Phytohormone Synthesis and Disease Resistance-Related Genes

Plant growth-promoting bacteria (PGPB) are components of the plant rhizosphere that promote plant growth and/or inhibit pathogen activity. To explore the cotton seedlings response to Bacillus circulans GN03 with high efficiency of plant growth promotion and disease resistance, a pot experiment was carried out, in which inoculations levels of GN03 were set at 104 and 108 cfu⋅mL–1. The results showed that GN03 inoculation remarkably enhanced growth promotion as well as disease resistance of cotton seedlings. GN03 inoculation altered the microbiota in and around the plant roots, led to a significant accumulation of growth-related hormones (indole acetic acid, gibberellic acid, and brassinosteroid) and disease resistance-related hormones (salicylic acid and jasmonic acid) in cotton seedlings, as determined with ELISA, up-regulated the expression of phytohormone synthesis-related genes (EDS1, AOC1, BES1, and GA20ox), auxin transporter gene (Aux1), and disease-resistance genes (NPR1 and PR1). Comparative genomic analyses was performed between GN03 and four similar species, with regards to phenotype, biochemical characteristics, and gene function. This study provides valuable information for applying the PGPB alternative, GN03, as a plant growth and disease-resistance promoting fertilizer.

Bacterization of forage grass seeds: Influence on spreading and growth of plants.

Inoculation of seeds before sowing of growth-stimulating microorganisms is an effective method for increasing the productivity of crops and forage grasses, in particular. In this work, nine strains of bacteria were tested for their ability to produce the phytohormone indolyl-3-acetic acid (IAA) and stimulate seed germination and elongation of shoots and roots of seedlings of mogar, sudanese grass, awnless brome, alfalfa, and meadow fescue. The highest IAA concentration in the culture liquid was found in the strains Enterobacter ludwigii BLK and Pseudomonas sp. UOM 10 (2001 and 1617 ng/ml, respectively), and the lowest was in Pseudomonas sp. 5GCH and P. koreensis IB-4 (144 and 141 ng/ml, respectively), however, at this stage of research, a direct relationship between the level of phytohormone synthesis and the ability of bacteria to positively affect plant growth was not found. The plants were characterized by the species specificity of the reaction to bacterial treatment, with the exception of the Pseudomonas sp. 5GCH, which in some cases had a positive effect on seed germination, but did not enhance the growth of roots and shoots in the studied herbs. Synchronous stimulation of seed germination and an increase in the length of shoots and roots was observed on mogar plants upon inoculation with E. ludwigii BLK and P. plecoglossicida CH5-2 and on an awnless brome (P. chlororaphis 4CH, B. vietnamiensis 8CH, E. ludwigii BLK, P. laurentiana ANT 17 and P. plecoglossicida CH5-2). None of the strains simultaneously increased the length of shoots and roots in Sudanese grass and meadow fescue, but most microorganisms enhanced seed germination in them. Bacterization did not increase germination in alfalfa, and only Pseudomonas sp. UOM 10 and P. koreensis IB-4 promoted the growth of its shoots and roots.

The intensity of ethylene release by soybean plants under the influence of fungicides in the early stages of legume-rhizobial symbiosis

The effect of pre-sowing treatment of soybean seeds with fungicides on the intensity of ethylene release, the processes of nodulation and nitrogen fixation in different symbiotic systems in the early stages of ontogenesis were investigated. The objects of the study were selected symbiotic systems formed with the participation of soybean (Glycine max (L.) Merr.) Diamond variety, strains Bradyrhizobium japonicum 634b (active, virulent) and 604k (inactive, highly virulent) and fungicides Maxim XL 035 PS (fludioxonil, 25 g/L, metalaxyl, 10 g/L), and Standak Top (fipronil, 250 g/L, thiophanate methyl, 225 g/L, piraclostrobin, 25 g/L). Before sowing, the seeds of soybean were treated with solutions of fungicides, calculated on the basis of one rate of expenditure of the active substance of each preparation indicated by the producer per ton of seed. One part of the seeds treated with fungicides was inoculated with rhizobium culture for 1 h (the titre of bacteria was 107 cells/mL). To conduct the research we used microbiological, physiological, biochemical methods, gas chromatography and spectrophotometry. It is found that, regardless of the effectiveness of soybean rhizobial symbiosis, the highest level of ethylene release by plants was observed in the stages of primordial leaf and first true leaf. This is due to the initial processes of nodulation – the laying of nodule primordia and the active formation of nodules on the roots of soybeans. The results show that with the participation of fungicides in different symbiotic systems, there are characteristic changes in phytohormone synthesis in the primordial leaf stage, when the nodule primordia are planted on the root system of plants. In particular, in the ineffective symbiotic system, the intensity of phytohormone release decreases, while in the effective symbiotic system it increases. At the same time, a decrease in the number of nodules on soybean roots inoculated with an inactive highly virulent rhizobia 604k strain due to the action of fungicides and an increase in their number in variants with co-treatment of fungicides and active virulent strain 634b into the stage of the second true leaf were revealed. It was shown that despite a decrease in the mass of root nodules, there is an increase in their nitrogen-fixing activity in an effective symbiotic system with the participation of fungicides in the stage of the second true leaf. The highest intensity of ethylene release in both symbiotic systems was recorded in the stage of the first true leaf, which decreased in the stage of the second true leaf and was independent of the nature of the action of the active substances of fungicides. The obtained data prove that the action of fungicides changes the synthesis of ethylene by soybean plants, as well as the processes of nodulation and nitrogen fixation, which depend on the efficiency of the formed soybean-rhizobial systems and their ability to realize their symbiotic potential under appropriate growing conditions.

Rhizosphere Metagenomics of Paspalum scrobiculatum L. (Kodo Millet) Reveals Rhizobiome Multifunctionalities

Multifunctionalities linked with the microbial communities associated with the millet crop rhizosphere has remained unexplored. In this study, we are analyzing microbial communities inhabiting rhizosphere of kodo millet and their associated functions and its impact over plant growth and survival. Metagenomics of Paspalum scrobiculatum L.(kodo millet) rhizopshere revealed taxonomic communities with functional capabilities linked to support growth and development of the plants under nutrient-deprived, semi-arid and dry biotic conditions. Among 65 taxonomically diverse phyla identified in the rhizobiome, Actinobacteria were the most abundant followed by the Proteobacteria. Functions identified for different genes/proteins led to revelations that multifunctional rhizobiome performs several metabolic functions including carbon fixation, nitrogen, phosphorus, sulfur, iron and aromatic compound metabolism, stress response, secondary metabolite synthesis and virulence, disease, and defense. Abundance of genes linked with N, P, S, Fe and aromatic compound metabolism and phytohormone synthesis—along with other prominent functions—clearly justifies growth, development, and survival of the plants under nutrient deprived dry environment conditions. The dominance of actinobacteria, the known antibiotic producing communities shows that the kodo rhizobiome possesses metabolic capabilities to defend themselves against biotic stresses. The study opens avenues to revisit multi-functionalities of the crop rhizosphere for establishing link between taxonomic abundance and targeted functions that help plant growth and development in stressed and nutrient deprived soil conditions. It further helps in understanding the role of rhizosphere microbiome in adaptation and survival of plants in harsh abiotic conditions.

A gall-forming insect manipulates hostplant phytohormone synthesis, concentrations, and signaling

Insect galls are highly specialized plant organs formed by an intimate biochemical interaction between the plant and a gall-inducing insect. Galls provide the insect enhanced nutrition and protection against natural enemies and environmental stresses. Because galls are plant organs, their development is likely to be governed by native phytohormones that function in organogenesis. We characterized concentrations of both growth and stress phytohormones in ungalled leaves and the leaf galls of Daktulosphaira vitifoliae on wild grape Vitis riparia at 4 different developmental stages. We found clear evidence of hormone manipulation by phylloxera during gall initiation and development. Phylloxera suppressed accumulation of of gibberellins, auxin, and jasmonates during the first two gall stages while absiscic acid concentrations were suppressed throughout development. Concentrations of 3 cytokinins and salicylate were greatly elevated during the earliest gall stage and declined sharply thereafter. We found no evidence of expression of cytokinin biosynthesis genes during the first gall stage, which strongly suggests that phylloxera supplied those phytohormones. High salicylate concentrations could have been caused by cytokinins, a response to microbes, or both. Our results suggest a central role for cytokinins in gall initiation and indicate the importance of the insect’s ability to manipulate other hormones.

Expression of Carotenoid Biosynthetic Genes and Carotenoid Biosynthesis during Seedling Development of Momordica charantia

Carotenoids belong to a large group of secondary metabolites, and have pivotal roles in plants, including photosynthesis and phytohormone synthesis, pigmentation, and membrane stabilization. Additionally, carotenoids are potent antioxidants, and their health benefits are becoming increasingly prominent. In recent years, carotenoids have been studied in many plants. Furthermore, gene expression, as well as carotenoid accumulation in different parts of the bitter melon, has been investigated; however, it has not been studied in bitter melon seedlings. In this study, carotenoid accumulation and transcript levels of McGGPPS1, McGGPPS2, McPSY, McPDS, McZDS, McLCYB, McLCYE1, McLCYE2, McCXHB, and McZEP, involved in carotenoid biosynthesis, were analyzed during seedling development using HPLC and qRT-PCR. The major carotenoids that accumulated in the bitter melon seedlings were lutein and E-β-carotene. The expression of most carotenoid biosynthetic genes increased during seedling development, consistent with the accumulation of violaxanthin, lutein, zeaxanthin, β-cryptoxanthin, 13Z-β-carotene, E-β-carotene, and 9Z-β-carotene in bitter melon seedlings. The results of this study provide a firm basis for comprehending the link between gene expression and carotenoid concentration in bitter melon seedlings.