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Ecological occurrence and plant regeneration of embryoid of the endangered and endemic plant Dysosma versipellis in China

In this study, the effective callus culture, somatic embryogenesis, and plant regeneration system of Dysosma versipellis, which is an endangered and endemic plant in China, were established under specific culture conditions. Using the D. versipellis leaves, petioles, and roots as explants, DPS software orthogonal design method and SPSS Duncans multiple range test were used to investigate their effects of D. versipellis on callus formation, embryoid induction, and plant regeneration by adding different phytohormones. Results showed that leaves and petioles were the most suitable materials in inducing callus. The effect of phytohormone on callus formation followed the order of 2,4-dichlorophenoxyacetic acid (2,4-D)>thidiazuron (TDZ)> kinetin>naphthylacetic acid (NAA)>2-ip. The best medium for callus formation was MS+2,4-D 1 mg/L+NAA 0.05 mg/L+TDZ 0.5 mg/L+2-ip 1 mg/L. The optimal medium to induce the formation of granular callus embryoid was MS+0.5 mg/L 6-BA+0.1 mg/L NAA, and the induction rate was 71.33%. The embryoid rooting and plant regeneration medium was MS+0.5 mg/L IBA+0.5 mg/L GA3. The optimal medium formula obtained in this study was suitable for the rapid induction of callus, embryoid, and plant regeneration of D. versipellis under in vitro culture conditions. Further study on the action mechanism, signal regulation mechanism, and artificial seed production of fungal elicitors affecting the accumulation of podophyllotoxin is important.

plant biology

Genome-wide association studies for yield-related traits in soft red winter wheat grown in Virginia

Grain yield is a trait of paramount importance in the breeding of all cereals. In wheat (Triticum aestivum L.), yield has steadily increased since the Green Revolution, though the current rate of increase is not forecasted to keep pace with demand due to growing world population and affluence. While several genome-wide association studies (GWAS) on yield and related component traits have been performed in wheat, the previous lack of a reference genome has made comparisons between studies difficult. In this study, a GWAS for yield and yield-related traits was carried out on a population of 324 soft red winter wheat lines across a total of four rain-fed environments in the state of Virginia using single-nucleotide polymorphism (SNP) marker data generated by a genotyping-by-sequencing (GBS) protocol. Two separate mixed linear models were used to identify significant marker-trait associations (MTAs). The first was a single-locus model utilizing a leave-one-chromosome-out approach to estimating kinship. The second was a sub-setting kinship multi-locus method (FarmCPU). The single-locus model identified nine significant MTAs for various yield-related traits, while the FarmCPU model identified 74 significant MTAs. The availability of the wheat reference genome allowed for the description of MTAs in terms of both genetic and physical positions, and enabled more extensive post-GWAS characterization of significant MTAs. The results indicate promising avenues for increasing grain yield by exploiting variation in traits relating to the number of grains per unit area, as well as phenological traits influencing grain-filling duration of genotypes.

plant biology

Effects of Biogas Slurry on Fruit Economic Traits and Soil Nutrient of Camellia oleifera Abel

Soil nutrients play a principal role in Camellia oleifera Abel (oil-seed camellia) production. Camellia oleifera absorbs nutrients from surrounding soils and its production is highly influenced by nutrients or fertilization. In this study, we investigated the effects of biogas slurry applications on soil nutrients and economic traits of C. oleifera fruits. Five different amounts of fertilizing biogas slurry (0, 10, 20, 30, or 40 kg/plant/year from three applications per year) were applied to C. oleifera plants in 2015 and 2016. Rhizosphere soil nutrients and C. oleifera fruit economic traits (yield, seed rate, and oil yield)were measured. Fertilization with biogas slurryincreasedsoil organic matter, available nitrogen (N), phosphorus (P), and potassium (K) in both 2015 and 2016. Increases in soil available N, P, and Kwere largest at the highest slurry application rate and second largest at the second highest application rate. Fruit economic traits were maximized at the two highest application rates. Oil yield was correlated withsoil available P in 2015 and 2016, and soil organic matter in 2015. Fertilization with biogas slurry decreased saturated fatty acid content in fruit but had no effect on unsaturated fatty acid content. In conclusion, fertilization with biogas slurry increases rhizosphere soil nutrients and fruit economic traits of C. oleifera with the rates of at least 30 kg/plant/year having the most positive effects.

plant biology

Transcriptome dynamics in developing leaves from C3 and C4 Flaveria species reveal determinants of Kranz anatomy

C4 species have evolved more than 60 times independently from C3 ancestors. This multiple and parallel evolution of the complex C4 trait indicates common underlying evolutionary mechanisms that might be identified by comparative analysis of closely related C3 and C4 species. Efficient C4 function depends on a distinctive leaf anatomy that is characterized by enlarged, chloroplast rich bundle sheath cells and a narrow vein spacing. To elucidate molecular mechanisms generating this so called Kranz anatomy, we analyzed a developmental series of leaves from the C4 plant Flaveria bidentis and the closely related C3 species Flaveria robusta using leaf clearing and whole transcriptome sequencing. Applying non-negative matrix factorization on the data identified four different zones with distinct transcriptome patterns in growing leaves of both species. Comparing these transcriptome patterns revealed an important role of auxin metabolism and especially auxin homeostasis for establishing the high vein density typical for C4 leaves.

plant biology

Sustaining yam yields amidst climate threat in the forest - savannah transition zone of Ghana

With about 70% of yam tuber been water, yield is critically affected during bulking as a result of onset of temporal drought. As a consequence of climate change, farmers who are into Dioscorea rotundata (white yam) production for local and international market lose their investments mainly due to erratic precipitation, drought spells culminating into low yields of just 12t/ha compared to the potential of about 22-49t/ha depending on the variety. Innovative land uses technologies with higher and sustained productivity for yam production are imperative. This study verifies improved agronomic package for sustainable yam production in yam growing areas in the forest - savannah transition zone of Ghana during the 2015 and 2016 cropping seasons. The improved agronomic package included use of ridging as seedbed, seed treatment before planting, fertilizer application at a rate of 30:30:36 N:P205:K20 kg/ha plus 15 kg/ha Mg and 20 kg/ha S as MgSO4 and the use of minimum stakes (trellis; 30-50% less number of stakes used by farmers staking). This was compared with farmers practice which consisted of mounding, no fertilizer application and no seed treatment. The results revealed significant (P [≤] 0.01) yam yields of more than 60% difference between the improved agronomic practice and farmers practice from Ejura, Atebubu and Kintampo yam growing communities. Adoption of improved agronomic practices does not only sustain yam production and address deforestation but also provide higher returns on investments promoting climate resilience by small holders.

plant biology

CRISPR-TSKO facilitates efficient cell type-, tissue-, or organ-specific mutagenesis in Arabidopsis

Detailed functional analyses of many fundamentally-important plant genes via conventional loss-of-function approaches are impeded by severe pleiotropic phenotypes. In particular, mutations in genes that are required for basic cellular functions and/or reproduction often interfere with the generation of homozygous mutant plants, precluding further functional studies. To overcome this limitation, we devised a CRISPR-based tissue-specific knockout system, CRISPR-TSKO, enabling the generation of somatic mutations in particular plant cell types, tissues, and organs. In Arabidopsis, CRISPR-TSKO mutations in essential genes caused well-defined, localized phenotypes in the root cap, stomatal lineage, or entire lateral roots. The underlying modular cloning system allows for efficient selection, identification, and functional analysis of mutant lines directly in the first transgenic generation. The efficacy of CRISPR-TSKO opens new avenues to discover and analyze gene functions in spatial and temporal contexts of plant life while avoiding pleiotropic effects of system-wide loss of gene function.

plant biology

The study of hormonal metabolism of Trincadeira and Syrah cultivars indicates new roles of salicylic acid, jasmonates, ABA and IAA during grape ripening and upon infection with Botrytis cinerea

Hormones play an important role in fruit ripening and in response to biotic stress. Nevertheless, analyses of hormonal profiling during plant development and defense are scarce. In this work, changes in hormonal metabolism in grapevine (Vitis vinifera) were compared between a susceptible (Trincadeira) and a tolerant (Syrah) variety during grape ripening and upon infection with Botrytis cinerea. Infection of grapes with the necrotrophic pathogen Botrytis cinerea leads to significant economic losses worldwide.\n\nPeppercorn-sized fruits were infected in the field and mock-treated and infected berries were collected at green, veraison and harvest stages for hormone analysis and targeted qPCR analysis of genes involved in hormonal metabolism and signaling. Results indicate a substantial reprogramming of hormonal metabolism during grape ripening and in response to fungal attack. Syrah and Trincadeira presented differences in the metabolism of abscisic acid (ABA), indole-3-acetic acid (IAA) and jasmonates during grape ripening that may be connected to fruit quality. On the other hand, high basal levels of salicylic acid (SA), jasmonates and IAA at an early stage of ripening, together with activated SA, jasmonates and IAA signaling, likely enable a fast defense response leading to grape resistance/ tolerance towards B. cinerea.\n\nThe balance among the different phytohormones seems to depend on the ripening stage and on the intra-specific genetic background and may be fundamental in providing resistance or susceptibility. In addition, this study indicated the involvement of SA and IAA in defense against necrotrophic pathogens and gains insights into possible strategies for conventional breeding and/or gene editing aiming at improving grape quality and grape resistance against Botrytis cinerea.

plant biology

Excess light priming in Arabidopsis thaliana with altered DNA methylomes

Plants must continuously react to the ever-fluctuating nature of their environment. Repeated exposure to stressful conditions can lead to priming, whereby prior encounters heighten a plants ability to respond to future events. A clear example of priming is provided by the model plant Arabidopsis thaliana (Arabidopsis), in which photosynthetic and photoprotective responses are enhanced following recurring light stress. While there are various post-translational mechanisms underpinning photoprotection, an unresolved question is the relative importance of transcriptional changes towards stress priming and, consequently, the potential contribution from DNA methylation - a heritable chemical modification of DNA capable of influencing gene expression. Here, we systematically investigate the potential molecular underpinnings of physiological priming against recurring excess-light (EL), specifically DNA methylation and transcriptional regulation: the latter having not been examined with respect to EL priming. The capacity for physiological priming of photosynthetic and photoprotective parameters following a recurring EL treatment was not impaired in Arabidopsis mutants with perturbed establishment, maintenance, or removal of DNA methylation. Importantly, no differences in development or basal photoprotective capacity were identified in the mutants that may confound the above result. Little evidence for a causal transcriptional component of physiological priming was identified; in fact, most alterations in primed plants presented as a transcriptional dampening in response to an additional EL exposure, likely a consequence of physiological priming. However, a set of transcripts uniquely regulated in primed plants provide preliminary evidence for a novel transcriptional component of recurring EL priming, independent of physiological changes. Thus, we propose that physiological priming of recurring EL in Arabidopsis occurs independently of DNA methylation; and that the majority of the associated transcriptional alterations are a consequence, not cause, of this physiological priming.\n\nOne sentence summaryPhotoprotection and priming against recurring excess light is functional despite impaired maintenance of the DNA methylome.

plant biology

The Flowering Hormone Florigen Accelerates Secondary Cell Wall Biogenesis to Harmonize Vascular Maturation with Reproductive Development

The protein hormone florigen is a universal systemic inducer of flowering and a generic growth terminator across meristems. To understand the developmental rational for its pleiotropic functions and to uncover the deep cellular systems mobilized by florigen beyond flowering we explored termination of radial expansion of stems. Employing the power of tomato genetics along with RNAseq and histological validations we show that endogenous, mobile, or induced florigen accelerates secondary cell wall biogenesis (SCWB), and hence vascular maturation, independently of flowering. This finding is supported by a systemic florigen antagonist from the non-flowering Ginkgo biloba, which arrests SCWB and by MADS and MIF genes downstream of florigen that similarly suppress or enhance, respectively, vascular maturation independent of flowering. We also show that florigen is remarkably stable and distributed to all organs regardless of existing endogenous levels. By accelerating SCWB, florigen reprograms the distribution of resources, signals and mechanical loads required for the ensuing reproductive phase it had originally set into motion.\n\nDevelopmental HighlightsO_LIFlorigen accelerates SCWB: A prime case for a long-range regulation of a complete metabolic network by a plant hormone.\nC_LIO_LIThe dual acceleration of flowering and vascular maturation by Florigen provides a paradigm for a dynamic regulation of global, independent, developmental programs.\nC_LIO_LIThe growth termination functions of florigen and the auto-regulatory mechanism for its production and distribution provide a communication network enveloping the shoot system.\nC_LIO_LIA stable florigen provides a possible mechanism for the quantitative regulation of flowering\nC_LIO_LILateral stimulation of xylem differentiation links the phloem-travelling florigen with the annual rings in trunks.\nC_LIO_LIMADS genes are common relay partners in Florigen circuits; vascular maturation in stems and reproductive transition in apical meristems.\nC_LI

plant biology

Physiological regulation of bud burst in grapevine

The physiological constraints on bud burst in woody perennials, including the prerequisite for vascular development remain unresolved. Both light and tissue oxygen status have emerged as important cues for vascular development in other systems, however, light requirement appears to be facultative in grapevine, and the information related to the spatial variability of oxygen in buds is unclear. Here, we analysed apoplastic development at early stages of grapevine bud burst and combined molecular modelling with histochemical techniques to determine the pore size of cell walls in grapevine buds. The data demonstrate that quiescent grapevine buds were impermeable to apoplastic dyes (acid fuchsin and eosin Y) until after bud burst was established. The molecular exclusion size was calculated to be 2.1 nm, which would exclude most macromolecules except simple sugars and phytohormones. In vivo experiments show that grapevine buds were able to resume growth even following excision from the cane, and that the outer scales of grapevine buds may participate in the biochemical repression of bud burst. Furthermore, we demonstrate that the tissue oxygen partial pressure data correlated well with structural heterogeneity within the bud and differences in tissue density. These data consolidate evidence that the meristematic core becomes rapidly oxygenated during bud burst. Taken together, and when put in the context of earlier studies, these data provide solid evidence that the physiological and biochemical events that initiate bud burst reside within the bud, and question the role of long distance signalling in this developmental transition.\n\nHighlightsO_LIThe apoplastic pore size between the grapevine bud and the mother vine is dynamically regulated in the transition to bud burst.\nC_LIO_LIThe molecular exclusion size of the apoplastic connection between the bud and cane is calculated 2.1 nm prior to the initiation of bud burst.\nC_LIO_LIThe structural heterogeneity of the bud explains the spatial variance in tissue oxygen status, and the meristematic core is oxygenated during the initiation of bud burst.\nC_LIO_LILong distance maternal signals are not a requirement for bud burst.\nC_LI

plant biology

A stress-response-related inter-compartmental signalling pathway regulates embryonic cuticle integrity in Arabidopsis

The embryonic cuticle is necessary for normal seed development and seedling establishment in Arabidopsis. Although mutants with defective embryonic cuticles have been identified, neither the deposition of cuticle material, nor its regulation, has been described during embryogenesis. Here we use electron microscopy, lipid staining and permeability assays to show that cuticle deposition initiates de novo in patches on globular embryos. By combining these techniques with genetics and gene expression analysis, we show that successful patch coalescence to form a continuous cuticle requires a signalling involving the endosperm-specific subtilisin protease ALE1 and the receptor kinases GSO1 and GSO2, which are expressed in the developing embryonic epidermis. Transcriptome analysis shows that this pathway regulates stress-related gene expression in seeds. Consistent with these findings we show genetically, and through activity analysis, that the stress-associated MPK6 protein acts downstream of GSO1 and GSO2 in the developing embryo. We propose that a stress-related signalling pathway has been hijacked in some angiosperm seeds through the recruitment of endosperm-specific components. Our work reveals the presence of an inter-compartmental dialogue between the endosperm and embryo that ensures the formation of an intact and functional cuticle around the developing embryo through an \"auto-immune\" type interaction.

plant biology

Population genome sequencing of the scab fungal species Venturia inaequalis, Venturia pirina, Venturia aucupariae and Venturia asperata.

The Venturia genus comprises fungal species that are pathogens on Rosaceae host plants, including V. inaequalis and V. asperata on apple, V. aucupariae on sorbus and V. pirina on pear. Although the genetic structure of V. inaequalis populations has been investigated in detail, genomic features underlying these subdivisions remain poorly understood. Here, we report whole genome sequencing of 87 Venturia strains that represent each species and each population within V. inaequalis. We present a PacBio genome assembly for the V. inaequalis EU-B04 reference isolate. The size of selected genomes was determined by flow cytometry, and varied from 45 to 93 Mb. Genome assemblies of V. inaequalis and V. aucupariae contain a high content of transposable elements (TEs), most of which belong to the Gypsy or Copia LTR superfamilies and have been inactivated by Repeat-Induced Point mutations. The reference assembly of V. inaequalis presents a mosaic structure of GC-equilibrated regions that mainly contain predicted genes and AT-rich regions, mainly composed of TEs. Six pairs of strains were identified as clones. Single-Nucleotide Polymorphism (SNP) analysis between these clones revealed a high number of SNPs that are mostly located in AT-rich regions due to misalignments and allowed determining a false discovery rate. The availability of these genome sequences is expected to stimulate genetics and population genomics research of Venturia pathogens. Especially, it will help understanding the evolutionary history of Venturia species that are pathogenic on different hosts, a history that has probably been substantially influenced by TEs.

plant biology

β-cyclocitric acid: a new apocarotenoid eliciting drought tolerance in plants

{beta}-Cyclocitral ({beta}-CC) is a volatile compound deriving from 1O2 oxidation of {beta}-carotene in plant leaves. {beta}-CC elicits a retrograde signaling, modulating 1O2-responsive genes and enhancing tolerance to photooxidative stress. Here, we show that {beta}-CC is largely converted into {beta}-cyclocitric acid ({beta}-CCA) in leaves and that this metabolite is a signal involved in stress tolerance. Treatment of Arabidopsis plants with {beta}-CCA markedly enhanced plant tolerance to drought by a mechanism different from known responses such as stomatal closure, changes in osmotic potential and jasmonate signaling. Furthermore, we show that the response to {beta}-CCA does not fully overlap with the {beta}-CC-dependent signaling, indicating that {beta}-CCA induces only a branch of the {beta}-CC signaling pathway. In addition, the protective effect of {beta}-CCA is a conserved mechanism, being observed in a variety of plant species. This study provides a new bioactive agent with promising agronomic applications for protecting plants against drought.

plant biology

HNI9 and HY5 maintain ROS homeostasis under high nitrogen provision in Arabidopsis

One sentence summaryExcessive N nutrition leads to ROS accumulation, and requires the function of major transcriptional regulators to maintain plants under physiological conditions.\n\nAuthor contributionsAn.M. and A.G. conceived research plans and supervised the experiments; F.B, Am.M., J.B., L.L., L.B. and An.M performed most of the experiments; F.B, Am.M., J.B., G.K., L.L., L.B. and An.M analyzed the data; An.M. wrote the article with contributions of all the authors.\n\nCompeting interestsThe authors declare no competing financial interests.\n\nSummaryReactive Oxygen Species (ROS) can accumulate in cells at excessive levels, leading to unbalanced redox status and to a potential oxidative stress, which can have damaging effects to the molecular components of plant cells. Several environmental conditions have been described as causing an elevation of ROS production in plants. Consequently, this requires the expression of detoxification responses in order to maintain ROS homeostasis at physiological levels. In case of mis-regulation of the detoxification systems, oxidative stress can lead ultimately to growth retardation and developmental defects. Here, we demonstrate that Arabidopsis plants growing under high nitrogen environment have to express a set of genes involved in detoxification of ROS in order to maintain ROS at physiological levels. We show that the chromatin factor HNI9 is an important actor of this response, required for the expression of these detoxification genes. Mutation in HNI9 leads to elevated ROS levels, and to ROS-dependent phenotypic defects under high but not low N provision. In addition, we identify HY5 as one of the major transcription factors also required for the expression of this detoxification program under high N condition. Our results demonstrate the requirement of a balance between N nutrition and ROS production, and identified the first major regulators required to control ROS homeostasis under excessive N nutrition.

plant biology

A SOSEKI-based coordinate system interprets global polarity cues in Arabidopsis

Multicellular development requires coordinated cell polarization relative to body axes, and translation to oriented cell division. In plants, it is unknown how cell polarities are connected to organismal axes and translated to division. Here, we identify Arabidopsis SOSEKI (SOK) proteins that integrate apical-basal and radial organismal axes to localize to polar cell edges. Localization does not depend on tissue context, requires cell wall integrity and is defined by a transferrable, protein-specific motif. SOK proteins structurally resemble the DIX oligomerization domain in the animal Dishevelled polarity regulator. The DIX-like domain self-interacts and is required for edge localization and for influencing division orientation. Our work identifies a plant compass, interpreted by SOK proteins. Furthermore, despite fundamental differences, polarity in plants and animals converge upon the same protein domain.

plant biology

Black pod disease profile: Monitoring its outbreak in Southwest, Nigeria

Black pod disease (BPD) has been and still remains a major threat to cocoa farmers worldwide due to its annual recurrence, fast spread and highly destructive nature. The disease has caused great anxiety in many cocoa producing communities due to the inability of indigenous cocoa farmers to determine when and where BPD outbreak will take place. Twelve (12) stations were structured from four important cocoa-producing States in the Southwestern region of Nigeria. An investigation of BPD outbreak was conducted in 2015/2016 within these regions. Infected cocoa pods and topsoil samples were collected for laboratory analysis. Pests attack, cherelle wilt and BPD outbreak were seasonal with 50% chances of occurrence in all the stations. Black pod diseases outbreak was recorded in all the States (100%) during the rainy season. The disease was at its peak in August 2015 in almost all the stations (station 1 (30.0%), station 3 (23.0%), station 11 (16.0%), station 4 (9.0%), station 5 (7.0%), and station 8 (3.0%). The height of disease severity was in September 2015 (station 1 (100.0%), station 3 (96.7%), station 5 (85.7%), station 11 (84.3%), and station 4 (70.0%), with station 8 reaching the 100% mark in October 2015. Most cocoa farmlands are now being abandoned, unless concerted efforts are made to effectively manage the disease, BPD will greatly reduce cocoa production in Nigeria and around the world.

plant biology

Partitioning index and non-structural carbohydrate dynamics among contrasting cassava genotypes under early terminal water stress

Cassava (Manihot esculenta Crantz.) is a storage root crop of importance in tropical regions where periodic dry season and drought affect performance. Cassava genotypes that differ in performance in ecosystems with various water regimes were subjected to water stress during storage-root initiation and early development. Plants were grown in 50 kg pots in a screen house environment under well-watered and water stress for over a 120-day period. Water stress had a significant effect on most traits analyzed. However, relative water content, partitioning index and non-structural carbohydrates were unaffected. Tolerant genotypes had a higher partitioning index than susceptible genotypes during water stress, associated with a larger number of storage roots initiated and larger storage root biomass, while they were shorter and had less fibrous root biomass. Tolerant lines were indistinguishable from susceptible lines in above ground biomass. These findings suggest that early evaluation of storage root number, partitioning index, and associated traits at an early stage of cassava storage-root development could be an effective approach by which cassava genotypes are screened for favorable drought tolerance response.

plant biology

Integrated multiomic analysis from chromatin to translation of stimulus-regulated gene activity exposes dominant patterns of nuclear-level control

Gene regulation is modulated from chromatin to translation. To better understand the integration of nuclear and cytoplasmic gene regulatory dynamics, we performed a multi-omic survey of the epigenome through the translatome of the response of Arabidopsis seedlings to hypoxia and reoxygenation. This included eight assays of chromatin (histones, accessibility, RNAPII and transcription factor binding) and three assays of RNA (nuclear, polyadenylated, and ribosome-associated). Dynamic patterns of nuclear regulation distinguished stress-induced and growth-associated mRNAs. The rapid upregulation of hypoxia-responsive gene transcripts and their preferential translation was accompanied by increased chromatin accessibility, RNAPII engagement and reduced Histone 2A.Z association. The more progressive upregulation of heat stress gene transcripts was characterized by early engagement of RNAPII and elevation of nuclear over polyadenylated RNA. Promoters of the rapidly versus progressively upregulated gene cohorts were enriched for cis-elements of ethylene-responsive and heat shock factor transcription factor families, respectively. By contrast, genes associated with growth including ribosomal proteins underwent distinct histone modifications, yet retained RNAPII engagement and accumulated nuclear transcripts during the stress. Upon reaeration, many of the progressively upregulated and growth-associated gene transcripts were mobilized to ribosomes. Thus, multi-level nuclear regulation distinguishes transcript synthesis, accumulation and translation in response to a transient stress.

plant biology