Tare Soil Disinfestation from Cyst Nematodes Using Inundation.

The dissemination of soil tares in the potato and sugar beet processing industry is one of the main paths for the spread of potato cyst nematodes (PCN), a severe quarantine pest. Efficient measures for the disinfestation of tare soil from PCN, but also from beet cyst nematodes (BCN), are needed. In our study, Globodera pallida (a PCN) and Heterodera schachtii (a BCN) cysts were sealed in gauze bags and imbedded in sedimentation basins. The cysts were either placed (a) in a presedimentation basin (Brukner basin) for three days, (b) in the presedimentation basin for three days and subsequently in sedimentation basins for nine weeks or (c) in sedimentation basins for nine weeks (without presedimentation). We tested the viability of the eggs and juveniles by hatching assays and using the reproduction rates in bioassays. We demonstrated that PCN and BCN imbedded in a sedimentation basin were only still showing some hatching activity after 2.5 weeks, while no hatching was observed when an additional Brukner basin treatment was conducted before sedimentation.

Selected Rhizosphere Bacteria Help Tomato Plants Cope with Combined Phosphorus and Salt Stresses.

Plants are often challenged by multiple abiotic stresses simultaneously. The inoculation of beneficial bacteria is known to enhance plant growth under these stresses, such as phosphorus starvation or salt stress. Here, for the first time, we assessed the efficiency of selected beneficial bacterial strains in improving tomato plant growth to better cope with double stresses in salty and P-deficient soil conditions. Six strains of Arthrobacter and Bacillus with different reservoirs of plant growth-promoting traits were tested in vitro for their abilities to tolerate 2-16% (w/v) NaCl concentrations, and shown to retain their motility and phosphate-solubilizing capacity under salt stress conditions. Whether these selected bacteria promote tomato plant growth under combined P and salt stresses was investigated in greenhouse experiments. Bacterial isolates from Cameroonian soils mobilized P from different phosphate sources in shaking culture under both non-saline and saline conditions. They also enhanced plant growth in P-deficient and salt-affected soils by 47-115%, and their PGP effect was even increased in higher salt stress conditions. The results provide valuable information for prospective production of effective bio-fertilizers based on the combined application of local rock phosphate and halotolerant phosphate-solubilizing bacteria. This constitutes a promising strategy to improve plant growth in P-deficient and salt-affected soils.

The Response of Maize to Inoculation with Arthrobacter sp. and Bacillus sp. in Phosphorus-Deficient, Salinity-Affected Soil.

Salinity and phosphorus (P) deficiency are among the most serious soil factors constraining crop productivity. A proposed strategy for alleviating these stresses is supporting plants by inoculation with growth-promoting rhizobacteria (PGPR). Here, a comparison of the ability of two maize composite and two F1 hybrid varieties to tolerate a P deficiency in either a saline or a non-saline environment showed that the uptake of nutrients by all four entries was significantly reduced by the imposition of both soil salinity and P deficiency, and that their growth was compromised to a similar extent. Subsequently, the ameliorative effect of inoculation with three strains of either Arthrobacter sp. or Bacillus sp. in an environment, which suffered simultaneously from salinity and P deficiency, was investigated. Inoculation with each of the strains was found to limit the plants' uptake of sodium cations, to increase their uptake of potassium cations, and to enhance their growth. The extent of the growth stimulation was more pronounced for the composite varieties than for the F1 hybrid ones, although the amount of biomass accumulated by the latter, whether the plants had been inoculated or not, was greater than that of the former varieties. When the bacterial strains were cultured in vitro, each of them was shown as able to produce the phytohormones auxin, abscisic acid, gibberellins, and cytokinins. The implication is that since the presence in the rhizospere of both Arthrobacter sp. and Bacillus sp. strains can support the growth of maize in salinity-affected and P deficient soils in a genotype-dependent fashion, it is important to not only optimize the PGPR strain used for inoculation, but also to select maize varieties which can benefit most strongly from an association with these bacteria.

Phage tail-like particles are versatile bacterial nanomachines - A mini-review.

Type VI secretion systems and tailocins, two bacterial phage tail-like particles, have been reported to foster interbacterial competition. Both nanostructures enable their producer to kill other bacteria competing for the same ecological niche. Previously, type VI secretion systems and particularly R-type tailocins were considered highly specific, attacking a rather small range of competitors. Their specificity is conferred by cell surface receptors of the target bacterium and receptor-binding proteins on tailocin tail fibers and tail fiber-like appendages of T6SS. Since many R-type tailocin gene clusters contain only one tail fiber gene it was appropriate to expect small R-type tailocin target ranges. However, recently up to three tail fiber genes and broader target ranges have been reported for one plant-associated Pseudomonas strain. Here, we show that having three tail fiber genes per R-type tailocin gene cluster is a common feature of several strains of Gram-negative (often plant-associated) bacteria of the genus Kosakonia. Knowledge about the specificity of type VI secretion systems binding to target bacteria is even lower than in R-type tailocins. Although the mode of operation implicated specific binding, it was only published recently that type VI secretion systems develop tail fiber-like appendages. Here again Kosakonia, exhibiting up to three different type VI secretion systems, may provide valuable insights into the antagonistic potential of plant-associated bacteria. Current understanding of the diversity and potential of phage tail-like particles is fragmentary due to various synonyms and misleading terminology. Consistency in technical terms is a precondition for concerted and purposeful research, which precedes a comprehensive understanding of the specific interaction between bacteria producing phage tail-like particles and their targets. This knowledge is fundamental for selecting and applying tailored, and possibly engineered, producer bacteria for antagonizing plant pathogenic microorganisms.

Data on molecular identification, phylogeny and in vitro characterization of bacteria isolated from maize rhizosphere in Cameroon.

Bacteria, which establish positive interactions with plant roots, play a key role in agricultural environments and are promising for their potential use in sustainable agriculture. Many of these mutualistic bacteria provide benefits to plant hosts by facilitating soil mineral nutrient uptake, protecting plants from biotic and abiotic stresses and producing substances that promote growth. The dataset presented here, is related to the publication entitled "Community structure and plant growth-promoting potential of cultivable bacteria isolated from Cameroon soil" (Tchuisseu et al., 2018) [1]. The data provide an extended analysis of the occurrence, taxonomical affiliation and functional traits of bacterial groups isolated from the rhizosphere of maize in Cameroon at different taxonomical levels, using a combination of molecular/bioinformatics tools and in vitro studies. Bacteria were isolated from maize rhizosphere soil. Isolated bacteria were identified using the 16s rRNA gene sequencing and phylogenetic analyses. All strains were characterized for their potential of salinity tolerance and growth promotion (phosphate solubilization, nifH gene presence and siderophore production) in order to select efficient bacterial strains for designing biological fertilizer exploitable for agriculture under specific stress conditions of the country. The data will be valuable for further studies on plant associated bacteria in Cameroon, which are still largely unexplored.

Comparative Genomics Reveal a Flagellar System, a Type VI Secretion System and Plant Growth-Promoting Gene Clusters Unique to the Endophytic Bacterium Kosakonia radicincitans.

The recent worldwide discovery of plant growth-promoting (PGP) Kosakonia radicincitans in a large variety of crop plants suggests that this species confers significant influence on plants, both in terms of yield increase and product quality improvement. We provide a comparative genome analysis which helps to unravel the genetic basis for K. radicincitans' motility, competitiveness and plant growth-promoting capacities. We discovered that K. radicincitans carries multiple copies of complex gene clusters, among them two flagellar systems and three type VI secretion systems (T6SSs). We speculate that host invasion may be facilitated by different flagella, and bacterial competitor suppression by effector proteins ejected via T6SSs. We found a large plasmid in K. radicincitans DSM 16656T, the species type strain, that confers the potential to exploit plant-derived carbon sources. We propose that multiple copies of complex gene clusters in K. radicincitans are metabolically expensive but provide competitive advantage over other bacterial strains in nutrient-rich environments. The comparison of the DSM 16656T genome to genomes of other genera of enteric plant growth-promoting bacteria (PGPB) exhibits traits unique to DSM 16656T and K. radicincitans, respectively, and traits shared between genera. We used the output of the in silico analysis for predicting the purpose of genomic features unique to K. radicincitans and performed microarray, PhyloChip, and microscopical analyses to gain deeper insight into the interaction of DSM 16656T, plants and associated microbiota. The comparative genome analysis will facilitate the future search for promising candidates of PGPB for sustainable crop production.

Plant growth-promoting bacteria Kosakonia radicincitans mediate anti-herbivore defense in Arabidopsis thaliana.

MAIN CONCLUSION: This study demonstrates that the application of the PGPB strain, Kosakonia radicincitans enhances a plant's resistance against phloem-feeding and chewing insects in Arabidopsis thaliana. The plant growth-promoting bacterial strain K. radicincitans DSM 16656 applied to A. thaliana reduced the number of phloem-feeding insects of both the specialist Brevicoryne brassicae and the generalist Myzus persicae. While weight gain of the generalist chewing insect Spodoptera exigua was reduced by 30% on A. thaliana plants treated with K. radicincitans, growth of the specialist caterpillar Pieris brassicae was not affected when compared with caterpillars from control plants. Since generalist and specialist chewing insects responded differentially to PGPB application, the implication of signaling pathways in PGPB mediated changes in plant defense was studied using two signaling pathway mutants impaired in their salicylic acid (npr1-1 mutant) or jasmonic acid (coi1-1 mutant) pathway. We found that the jasmonic acid pathway is relevant for upregulation of aliphatic glucosinolates and suppression of the chewing generalist S. exigua larval growth. Chewing from generalist P. brassicae increased glucosinolate content in A. thaliana leaves mediated via both signaling pathways. However, only in the npr1-1 mutant, which contains the highest aliphatic glucosinolate content, the P. brassicae induced further enrichment of glucosinolates, resulting in a reduction of larval growth. Effects of K. radicincitans on plant resistance could not be explained by changes in glucosinolate contents or composition. Our results demonstrate the distinct role played by K. radicincitans in suppressing insect performance in A. thaliana.

Community structure and plant growth-promoting potential of cultivable bacteria isolated from Cameroon soil.

Exploiting native plant growth-promoting rhizobacteria (PGPR) in Cameroonian agro-ecosystems provides a means to improve plant-microbe interactions that may enhance ecosystem sustainability and agricultural productivity in an environmentally eco-friendly way. Consequently, we aimed to investigate the community structure and functional PGPR diversity of maize grown in Cameroon. Native bacteria isolated from Cameroon maize rhizosphere soil were identified by partial 16S rRNA gene sequencing and screened for traits particularly relevant for Cameroon low-fertility soil conditions, such as their abilities to tolerate high concentrations of salt, and their plant growth- promoting potential. Genetic and functional diversity was characterized according to their phylogenetic affiliation. A total of 143 bacteria were identified and assigned to 3 phyla (Actinobacteria, Firmicutes and Proteobacteria), 13 families and 20 genera. Bacillus (31.5%), Arthrobacter (17.5%), and Sinomonas (13.3%) were the most abundant genera identified among all the isolates. Based on their in vitro characterization, 88.1% were salt tolerant at 2% NaCl, but only 16.8% could tolerate 8% NaCl, 50.4% solubilized phosphate, 10.5% possessed the nifH gene, and 19.6% produced siderophores. Six isolates affiliated to the most abundant genera identified in this work, Bacillus and Arthrobacter, carrying multiple or only single tested traits were selected to evaluate their growth- promoting potential in an in vitro maize germination assay. Three strains possessing multiple traits induced significantly increased hypocotyl and root length of maize seeds compared to non-inoculated control seeds. Our results indicate the potential of selected indigenous Cameroon rhizobacteria to enhance maize growth.

Successful Formulation and Application of Plant Growth-Promoting Kosakonia radicincitans in Maize Cultivation.

The global market for biosupplements is expected to grow by 14 percent between 2014 and 2019 as a consequence of the proven benefits of biosupplements on crop yields, soil fertility, and fertilizer efficiency. One important segment of biosupplements is plant growth-promoting bacteria (PGPB). Although many potential PGPB have been discovered, suitable biotechnological processing and shelf-life stability of the bacteria are challenges to overcome for their successful use as biosupplements. Here, the plant growth-promoting Gram-negative strain Kosakonia radicincitans DSM 16656T (family Enterobacteriaceae) was biotechnologically processed and applied in the field. Solid or liquid formulations of K. radicincitans were diluted in water and sprayed on young maize plants (Zea mays L.). Shelf-life stability tests of formulated bacteria were performed under 4°C and -20°C storage conditions. In parallel, the bacterial formulations were tested at three different farm level field plots characterized by different soil properties. Maize yield was recorded at harvest time, and both formulations increased maize yields in silage as well as grain maize, underlining their positive impact on different agricultural systems. Our results demonstrate that bacteria of the family Enterobacteriaceae, although incapable of forming spores, can be processed to successful biosupplements.

The plant growth-promoting bacterium Kosakonia radicincitans improves fruit yield and quality of Solanum lycopersicum.

BACKGROUND: Production and the quality of tomato fruits have a strong economic relevance. Microorganisms such as the plant growth-promoting bacterium (PGPB) Kosakonia radicincitans (DSM 16656) have been demonstrated to improve shoot and root growth of young tomato plants, but data on yield increase and fruit quality by K. radicincitans are lacking. RESULTS: This study investigated how K. radicincitans affects tomato fruits. After inoculation of tomato seeds with K. radicincitans or a sodium chloride buffer control solution, stalk length, first flowering and the amount of ripened fruits produced by inoculated and non-inoculated plants were monitored over a period of 21 weeks. Inoculation of tomato seeds with K. radicincitans accelerated flowering and ripening of tomato fruits. Sugars, acidity, amino acids, volatile organic compounds and carotenoids in the fruits were also analyzed. CONCLUSION: It was found that the PGPB K. radicincitans affected the amino acid, sugar and volatile composition of ripened fruits, contributing to a more pleasant-tasting fruit without forfeiting selected quality indicators. © 2017 Society of Chemical Industry.

Health-related quality of life among women in rural Bangladesh after surgical repair of obstetric fistula.

OBJECTIVE: To identify factors influencing health-related quality of life (QOL) among women in Bangladesh after successful surgical repair of obstetric fistula. METHODS: In 2012, a cross-sectional study was conducted of women who had undergone a successful surgical repair of an obstetric fistula at a hospital in northwest Bangladesh between June 2005 and May 2012. Sociodemographic, obstetric, and fistula-related information was collected by interview. Health-related QOL was assessed using the 36-item Short Form Health Survey (SF-36). RESULTS: A total of 113 women were included. Linear regression analysis showed that discrimination experiences negatively affected four of the eight scales of SF-36, as well as both the physical and mental component summaries (P<0.05 for all). Married status had a positive effect on four scales and the mental component summary (P<0.05 for all). Age inversely affected two scales and the physical component summary (P<0.01 for all). Internalized stigma negatively influenced two scales (P<0.05 for both). Not having living children and experiencing two or more previous stillbirths negatively influenced one scale (P<0.05 for both). CONCLUSION: Several factors influence health-related QOL after obstetric fistula repair; a comprehensive approach is needed to address these issues.

Impact of the PGPB Enterobacter radicincitans DSM 16656 on growth, glucosinolate profile, and immune responses of Arabidopsis thaliana.

Plant growth-promoting bacteria (PGPB) affect plant cellular processes in various ways. The endophytic bacterial strain Enterobacter radicincitans DSM 16656 has been shown to improve plant growth and yield in various agricultural and vegetable crops. Besides its ability to fix atmospheric nitrogen, produce phytohormones, and solubilize phosphate compounds, the strain is highly competitive against native endophytic organisms and colonizes the endorhizosphere in high numbers. Here, we show that E. radicincitans inoculation of the noncrop plant Arabidopsis thaliana promotes plant growth. Furthermore, high performance liquid chromatography (HPLC) analysis revealed that bacterial inoculation slightly decreased amounts of aliphatic glucosinolates in plant leaves in a fast-growing stage but increased these compounds in an older phase where growth is mostly completed. This effect seems to correlate with developmental stage and depends on the nitrogen requirement. Additionally, nitrogen deficiency studies with seedlings grown on medium containing different nitrogen concentrations suggest that plant nitrogen demand can influence the intensity of plant growth enhancement by E. radicincitans. This endophyte seems not to activate stress-inducible mitogen-activated protein kinases (MAPKs). Analyzing transcription of the defense-related genes PR1, PR2, PR5, and PDF1.2 by quantitative real time polymerase chain reaction (qPCR) revealed that E. radicincitans DSM 16656 is able to induce priming via salicylic acid (SA) or jasmonate (JA)/ethylene (ET) signaling pathways to protect plants against potential pathogen attack.

Jasmonate and ppHsystemin regulate key Malonylation steps in the biosynthesis of 17-Hydroxygeranyllinalool Diterpene Glycosides, an abundant and effective direct defense against herbivores in Nicotiana attenuata.

We identified 11 17-hydroxygeranyllinalool diterpene glycosides (HGL-DTGs) that occur in concentrations equivalent to starch (mg/g fresh mass) in aboveground tissues of coyote tobacco (Nicotiana attenuata) and differ in their sugar moieties and malonyl sugar esters (0-2). Concentrations of HGL-DTGs, particularly malonylated compounds, are highest in young and reproductive tissues. Within a tissue, herbivore elicitation changes concentrations and biosynthetic kinetics of individual compounds. Using stably transformed N. attenuata plants silenced in jasmonate production and perception, or production of N. attenuata Hyp-rich glycopeptide systemin precursor by RNA interference, we identified malonylation as the key biosynthetic step regulated by herbivory and jasmonate signaling. We stably silenced N. attenuata geranylgeranyl diphosphate synthase (ggpps) to reduce precursors for the HGL-DTG skeleton, resulting in reduced total HGL-DTGs and greater vulnerability to native herbivores in the field. Larvae of the specialist tobacco hornworm (Manduca sexta) grew up to 10 times as large on ggpps silenced plants, and silenced plants suffered significantly more damage from herbivores in N. attenuata's native habitat than did wild-type plants. We propose that high concentrations of HGL-DTGs effectively defend valuable tissues against herbivores and that malonylation may play an important role in regulating the distribution and storage of HGL-DTGs in plants.

Silencing the hydroxyproline-rich glycopeptide systemin precursor in two accessions of Nicotiana attenuata alters flower morphology and rates of self-pollination.

Systemins and their hydroxyproline-rich glycopeptide systemin (ppHS) subfamily members are known to mediate antiherbivore defenses in some solanaceous taxa but not others; functions other than in defense remain largely unexplored. Nicotiana attenuata's ppHS is known not to function in herbivore defense. NappHS transcripts are abundant in flowers, particularly in pistils, and when two N. attenuata accessions from Utah and Arizona were transformed to silence NappHS by RNAi (IRsys), seed capsule production and seed number per capsule were reduced in both accessions. These reductions in reproductive performance could not be attributed to impaired pollen or ovule viability; hand-pollination of all IRsys lines of both accessions restored seed production per capsule to levels found in wild-type plants. Rather, changes in flower morphology that decreased the efficiency of self-pollination are likely responsible: IRsys plants of both accessions have flowers with pistils that protrude beyond their anthers. Because these changes in flower morphology are reminiscent of CORONATINE-INSENSITIVE1-silenced N. attenuata plants, we measured jasmonates (JAs) and their biosynthetic transcripts in different floral developmental stages, and found levels of JA-isoleucine (Ile)/leucine and threonine deaminase transcripts, which are abundant in wild-type pistils, to be significantly reduced in IRsys buds and flowers. Threonine deaminase supplies Ile for JA-Ile biosynthesis, and we propose that ppHS mediates JA signaling during flower development and thereby changes flower morphology. These results suggest that the function of ppHS family members in N. attenuata may have diversified to modulate flower morphology and thereby outcrossing rates in response to biotic or abiotic stresses.

The hydroxyproline-rich glycopeptide systemin precursor NapreproHypSys does not play a central role in Nicotiana attenuata's anti-herbivore defense responses.

To determine whether the Nicotiana tabacum preproHypSys homolog in Nicotiana attenuata (NapreproHypSys) mediates anti-herbivore responses, we silenced (IRsys) and ectopically over-expressed (OVsys) NapreproHypSys in N. attenuata. Neither herbivore simulation nor methyl jasmonate (MeJA) application increased transcripts in wild-type (WT) or transformed lines. Compared to WT plants, OVsys plants had marginally higher constitutive levels but normally induced levels of trypsin proteinase inhibitors (TPIs) and nicotine; IRsys plants did not differ from WT plants. Herbivory-associated signalling [salicylic acid-induced protein kinase (SIPK) activity, jasmonic acid (JA), jasmonic acid-isoleucine/leucine (JA-Ile/Leu) and ethylene production or perception] did not differ strongly among the lines, but JA, JA-Ile/Leu and ethylene were marginally higher in OVsys plants. Manduca sexta larval performance did not differ among the lines, but feeding induced levels of TPI and nicotine in OVsys plants and decreased them in IRsys plants relative to WT. The secondary metabolite profiles of plants transplanted into N. attenuata's native habitat in the Great Basin Desert (UT, USA) mirrored those of glasshouse-grown plants, and compared to WT plants, OVsys plants suffered marginally less damage from grasshoppers, mirids and flea beetles but did not differ in their ability to attract Geocoris predators. We conclude that NapreproHypSys does not play a central role in anti-herbivore defense signalling in this native tobacco.

Occurrence of tetraploidy in Nicotiana attenuata plants after Agrobacterium-mediated transformation is genotype specific but independent of polysomaty of explant tissue.

Genotypes of Nicotiana attenuata collected from Utah and Arizona were transformed with 17 different vectors (14 unpublished vectors based on 3 new backbone vectors) using an Agrobacterium-mediated procedure to functionally analyze genes important for plant-insect interactions. None of the 51 T1-T3 transgenic Utah lines analyzed by the flow cytometry were tetraploid, as opposed to 18 of 33 transgenic Arizona lines (55%). Analysis of T0 regenerants transformed with the same vector carrying an inverted repeat (IR) N. attenuata pro-systemin construct confirmed the genotype dependency of tetraploidization: none of the 23 transgenic Utah lines were tetraploid but 31 (72%) of 43 transgenic Arizonas were tetraploid. We tested the hypothesis that the differences in polysomaty of the explant tissues accounted for genotype dependency of tetraploid formation by measuring polysomaty levels in different seedling tissues. Hypocotyls, cotyledons, and roots of Utah and Arizona genotypes contained similar percentages of 4C nuclei (61 and 60; 7 and 5; and 58 and 61%, respectively). Since we used hypocotyls as explant sources and the nonoccurrence of tetraploid Utah transformants does not correspond to the high percentage of 4C nuclei in Utah hypocotyls, we can rule out a direct relationship between tetraploid formation and polysomaty level. We hypothesize that the difference between the Utah and Arizona genotypes results from the failure of polyploid Utah callus to regenerate into fully competent plants. We propose that future work on post-transformation polyploidy concentrate on the processes that occur during callus formation and plant regeneration from callus.