Using FIBexDB for In-Depth Analysis of Flax Lectin Gene Expression in Response to Fusarium oxysporum Infection

Plant proteins with lectin domains play an essential role in plant immunity modulation, but among a plurality of lectins recruited by plants, only a few members have been functionally characterized. For the analysis of flax lectin gene expression, we used FIBexDB, which includes an efficient algorithm for flax gene expression analysis combining gene clustering and coexpression network analysis. We analyzed the lectin gene expression in various flax tissues, including root tips infected with Fusarium oxysporum. Two pools of lectin genes were revealed: downregulated and upregulated during the infection. Lectins with suppressed gene expression are associated with protein biosynthesis (Calreticulin family), cell wall biosynthesis (galactose-binding lectin family) and cytoskeleton functioning (Malectin family). Among the upregulated lectin genes were those encoding lectins from the Hevein, Nictaba, and GNA families. The main participants from each group are discussed. A list of lectin genes, the expression of which can determine the resistance of flax, is proposed, for example, the genes encoding amaranthins. We demonstrate that FIBexDB is an efficient tool both for the visualization of data, and for searching for the general patterns of lectin genes that may play an essential role in normal plant development and defense.

Survival of Plants During Short-Term BOA-OH Exposure: ROS Related Gene Expression and Detoxification Reactions Are Accompanied With Fast Membrane Lipid Repair in Root Tips

For the characterization of BOA-OH insensitive plants, we studied the time-dependent effects of the benzoxazolinone-4/5/6/7-OH isomers on maize roots. Exposure of Zea mays seedlings to 0.5 mM BOA-OH elicits root zone-specific reactions by the formation of dark rings and spots in the zone of lateral roots, high catalase activity on root hairs, and no visible defense reaction at the root tip. We studied BOA-6-OH- short-term effects on membrane lipids and fatty acids in maize root tips in comparison to the benzoxazinone-free species Abutilon theophrasti Medik. Decreased contents of phosphatidylinositol in A. theophrasti and phosphatidylcholine in maize were found after 10–30 min. In the youngest tissue, α-linoleic acid (18:2), decreased considerably in both species and recovered within one hr. Disturbances in membrane phospholipid contents were balanced in both species within 30–60 min. Triacylglycerols (TAGs) were also affected, but levels of maize diacylglycerols (DAGs) were almost unchanged, suggesting a release of fatty acids for membrane lipid regeneration from TAGs while resulting DAGs are buildings blocks for phospholipid reconstitution, concomitant with BOA-6-OH glucosylation. Expression of superoxide dismutase (SOD2) and of ER-bound oleoyl desaturase (FAD2-2) genes were contemporaneously up regulated in contrast to the catalase CAT1, while CAT3 was arguably involved at a later stage of the detoxification process. Immuno-responses were not elicited in short-terms, since the expression of NPR1, POX12 were barely affected, PR4 after 6 h with BOA-4/7-OH and PR1 after 24 h with BOA-5/6-OH. The rapid membrane recovery, reactive oxygen species, and allelochemical detoxification may be characteristic for BOA-OH insensitive plants.

Clinical insights into the three-dimensional anatomy of cheek teeth in alpacas based on micro-computed tomography - Part 2: Maxillary cheek teeth

Background Scientific knowledge regarding alpaca dentistry is relatively limited despite its clinical implications. The present gap in available supportive data leads to limited treatment options for dental pathology in alpacas in comparison to other species. The main goal of this study was to gain novel insights into the general and pulp morphology of maxillary cheek teeth to allow development of more advanced treatment strategies in the future. Also, the risk of causing pulp exposure when floating maxillary cheek teeth was of particular interest. Concurent research focusing on the anatomy of mandibular cheek teeth has been performed accordingly. The results obtained in mandibular teeth are expected to be non-extrapolatable because of the structural differences between mandibular and maxillary teeth. Results Pulp morphology of maxillary cheek teeth showed great variation. A common pulp chamber was identified in 46/83 (55.4%) teeth with a mean dental age of 2 years and 7 months (± 2 years and 5 months). Pulpal segmentation was more commonly observed in teeth of increasing age. Full columnar segmentation was seen in 33/69 teeth (47.8%), whereas within-column segmentation was observed in 36/83 teeth (43.4%). Age and degree of segmentation of the pulpal tissue varied greatly according to Triadan position. Physical contact between roots of adjacent teeth was found in the majority of examined molars (range 82–94%) which resulted in morphological adaptations at the level of the root tips. The measured sub-occlusal dentinal thickness was as low as 0.46 mm above pulp horn 2 in a 14 years and 11 months old Triadan 09, emphasizing the risk of pulp exposure attributed to dental floating. Conclusion This study offers an objective description of age-dependent maxillary cheek teeth pulp morphology in alpacas. Current findings are of great value to provide a basis for the development of tooth-saving techniques as a treatment for dental disease in this species. Observed physical contact between the roots of different examined molars may be a facilitating factor in the spread of apical infection in chronically diseased cases. Finally, a conservative approach regarding dental floating is recommended in order to avoid iatrogenic damage to pulp tissue.

Analysis of Hypericum accessions by DNA fingerprinting and flow cytometry

Hypericum perforatum, H. umbellatum, H. maculatum, and H. hircinum accessions originating from botanical gardens across Europe were examined by flow cytometry and molecular markers. 2C DNA content of 17 Hypericum perforatum accessions (Hp) and the H. perforatum cultivar Topaz amounted to between 1.56 pg and 1.62 pg. In four Hp accessions some individual plants were found with a DNA content corresponding to 6Cx (2.34 - 2.39 pg). All plants of accession Hp8 showed a DNA content of 6Cx (2.41 pg). In root tips of Hp plants with an average DNA amount of 1.58 pg, 32 chromosomes were detected, corresponding to 2n = 4x. This is the first ploidy and/or DNA content report for H. umbellatum, H. maculatum and H. hircinum. H. umbellatum and H. maculatum, each contained 0.76 pg DNA and 16 chromosomes were counted. The 2C DNA content of H. hircinum was 1.00 pg with the best metaphase plate revealing 32 chromosomes. Additionally, a combined marker analysis, based on inter-simple sequence repeats (ISSR) and sequence related amplified polymorphism (SRAP), was conducted to gain a better understanding of diversity especially within the accessions of H. perforatum. A total of 27 (11 ISSR and 16 SRAP) primer combinations were screened, showing 699 bands, of which 661 were polymorphic. UPGMA clustering revealed that accessions from the same geographic area tended to be more closely related, while H. maculatum was grouped separately from all H. perforatum accessions. Both methods have shown similar sensitivities in detecting the genetic diversity of the analyzed genotypes. Our results may be useful for Hypericum breeding programs and the development of effective conservation strategies.

Physiological and Transcriptomic Characterization of Sea-Wheatgrass-Derived Waterlogging Tolerance in Wheat

Waterlogging, causing hypoxia stress and nitrogen depletion in the rhizosphere, has been an increasing threat to wheat production. We developed a wheat–sea wheatgrass (SWG) amphiploid showing superior tolerance to waterlogging and low nitrogen. Validated in deoxygenated agar medium for three weeks, hypoxia stress reduced the dry matter of the wheat parent by 40% but had little effect on the growth of the amphiploid. To understand the underlying mechanisms, we comparatively analyzed the wheat–SWG amphiploid and its wheat parent grown in aerated and hypoxic solutions for physiological traits and root transcriptomes. Compared with its wheat parent, the amphiploid showed less magnitude in forming root porosity and barrier to radial oxygen loss, two important mechanisms for internal O2 movement to the apex, and downregulation of genes for ethylene, lignin, and reactive oxygen species. In another aspect, however, hypoxia stress upregulated the nitrate assimilation/reduction pathway in amphiploid and induced accumulation of nitric oxide, a byproduct of nitrate reduction, in its root tips, and the amphiploid maintained much higher metabolic activity in its root system compared with its wheat parent. Taken together, our research suggested that enhanced nitrate assimilation and reduction and accumulation of nitric oxide play important roles in the SWG-derived waterlogging tolerance.

Diversity of ectomycorrhizal fungi associated with Quercus petraea in the national park Fruška gora in the Republic of Serbia

Although sessile oak is one of the most important deciduous forest tree species in Europe, data on the diversity of ectomycorrhizal (ECM) fungi on sessile oaks in the Republic of Serbia are scarce. The aim of this study was to provide the first insight into the diversity of ECM fungi on sessile oak in Serbia. Two sites Info center and Brankovac, located in National Park Fruška gora were chosen. ECM fungi were identified combining morphological and anatomical characterization with molecular analysis of nuclear rDNA internal transcribed spacer (ITS) region. All vital ECM root tips were counted, diversity indices were calculated, and ECM fungi were classified into the exploration types. The granulometric and chemical composition of soil were analyzed as well. At both sites, 26 different ECM fungal taxa were recorded in total, 17 taxa were observed at the site Info center and 12 taxa at Brankovac. ECM communities consisted of a few abundant taxa and a larger number of rare taxa. Lactarius quietus, Cenococcum geophilum, and Tomentella sublilacina were recorded at both sites. High abundance of contact and short-distance exploration types recorded in studied stands suggests that soils are sufficiently rich in total nitrogen and organic matter. Values of diversity indices recorded in studied sessile oak stands from Fruška gora were lower in comparison to those obtained in stands of different oak species across Europe which is likely induced by drought. To get a more thorough insight into the diversity of ECM fungi on sessile oak, research should be continued at more sites and seasonal dynamics should be included.

Three Antimitotic Compounds Found in Banana Pseudo-Stem By Using Earthworms.

To identify antimitotic compounds from abundant and inexpensive plant resources, banana pseudo-stem was (BPS) chosen. Onion root tip assay and earthworm regeneration assay were carried out to test theantimitotic potential of aqueous extract of BPS.Earthworm (Eudrilus eugeniae) regeneration assay exploits the regeneration ability of amputated earthworms that retain the clitellum region. Aqueous extract of BPS decreased the mitotic index in Allium cepa root tips. Besides, thisaqueous extract of BPS inhibited the regeneration of blastema from amputated earthworms as well. Validation of this extract with MTT (3-(4,5-dimethyl thiazolyl-2-yl)- 2,5-diphenyltetrazolium bromide) assay using MCF-7 breast cancer cell linefurther supported the presence of antimitotic compounds. Aqueous BPS extract was further fractionated with ethyl acetateand it was found to inhibit the regeneration of new tissues from amputated earthworms. Liquid Chromatography and Mass spectrometry (LC-MS) analysis was performed with aqueousBPS extract to predict the lead compounds.Prediction analysis with mass values revealed the presence of three different compounds viz. α-tocotrienol, 1,2,4-nonadecanetriol and 3',4',7-trihydroxyisoflavone, which were already reported to inhibit the cell division. All our results strongly supported that banana pseudo-stem extract possesses antimitotic compounds. This is the first report of identification of putative antimitotic compounds from aqueous extract of Musa paradisiaca var. Robusta by using earthworms.

Whole-Tissue Three-Dimensional Imaging of Rice at Single-Cell Resolution

The three-dimensional (3D) arrangement of cells in tissues provides an anatomical basis for analyzing physiological and biochemical aspects of plant and animal cellular development and function. In this study, we established a protocol for tissue clearing and 3D imaging in rice. Our protocol is based on three improvements: clearing with iTOMEI (clearing solution suitable for plants), developing microscopic conditions in which the Z step is optimized for 3D reconstruction, and optimizing cell-wall staining. Our protocol successfully 3D imaged rice shoot apical meristems, florets, and root apical meristems at cellular resolution throughout whole tissues. Using fluorescent reporters of auxin signaling in rice root tips, we also revealed the 3D distribution of auxin signaling events that are activated in the columella, quiescent center, and multiple rows of cells in the stele of the root apical meristem. Examination of cells with higher levels of auxin signaling revealed that only the central row of cells was connected to the quiescent center. Our method provides opportunities to observe the 3D arrangement of cells in rice tissues.

Genetic Variation for Traits Related to Phosphorus Use Efficiency in Lens Species at the Seedling Stage

Phosphorus (P) is an essential, non-renewable resource critical for crop productivity across the world. P is immobile in nature and, therefore, the identification of novel genotypes with efficient P uptake and utilization under a low P environment is extremely important. This study was designed to characterize eighty genotypes of different Lens species for shoot and root traits at two contrasting levels of P. A significant reduction in primary root length (PRL), total surface area (TSA), total root tips (TRT), root forks (RF), total dry weight (TDW), root dry weight (RDW) and shoot dry weight (SDW) in response to P deficiency was recorded. A principal component analysis revealed that the TDW, SDW and RDW were significantly correlated to P uptake and utilization efficiency in lentils. Based on total dry weight (TDW) under low P, L4727, EC718309, EC714238, PL-97, EC718348, DPL15, PL06 and EC718332 were found promising. The characterization of different Lens species revealed species-specific variations for the studied traits. Cultivated lentils exhibited higher P uptake and utilization efficiency as compared to the wild forms. The study, based on four different techniques, identified EC714238 as the most P use-efficient genotype. The genotypes identified in this study can be utilized for developing mapping populations and deciphering the genetics for breeding lentil varieties suited for low P environments.

Growth promotion of raspberry and strawberry plants by bacterial inoculants

Study on potential mechanisms influencing the growth of raspberry and strawberry plants showed that the most active was Bacillus sp. strain AF75BC producing IAA and siderophores, and having the ability to release phosphorus. The latter feature was also present in the strains Sp115AD (B. subtilis) and SP116AC (Paenibacillus polymyxa). Two of the tested strains: SP116AC and JaFGU (Lysobacter sp.) showed the ability to fix atmospheric nitrogen, while the AF75AB2 (Bacillus sp.) produced siderophores and IAA. All strains showed an antagonism toward the most important pathogens of strawberry and raspberry, i.e. Verticillium dahliae, Botrytis cinerea, Phytophthora cactorum and Colletotrichum acutatum, limiting their growth to a different extent on the PDA medium. Inoculation of raspberry roots with the tested bacteria resulted in an increase of some growth parameters of their above-ground part in cv. Poemat. In the case of cv. Polana, a significant increase was found only in the chlorophyll content in the leaves. All the inoculants caused an increase in dry mass of roots in cv. Polana, and in cv. Poemat similar effect was observed after applying Inoculants 1 and 3. The treatments of strawberry roots with any of the inoculants resulted in a significant increase in the total leaf surface area in cv. Rumba, but they had no effect on the chlorophyll content in the leaves of either cultivar. All the inoculants significantly increased the total length of roots and their total surface area in cv. Rumba. This parameter also increased in cv. Elsanta, and the number of root tips also significantly increased in this cultivar. Our study showed that the tested inocula is a promising alternative as a bio-fertilizer for small fruit production in sustainable and organic agricultural systems.