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Search indexed bioRxiv preprints in genomics, neuroscience, cell biology and bioinformatics. Read source abstracts and check manuscript versions; preprints are not peer reviewed.

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bioRxiv2017

Parsing Of Compositions And Microstructure Characteristics For Rust-Spots Of Pear Pericarp

The commercial value of Kurles pears pericarp, a popular and favored fruit for its unique aroma and refreshingly crisp texture, had sharply decreased because of a rust breakout of the beloved pear in China during the atmosphere-controlled storage. High performance liquid chromatography (HPLC) and liquid chromatography-mass spectrometry (LC-MS) were used to analyze rust spots on the pericarp of Kurle pears. Therefore, the chemical compounds of four various eluates, originating from the rust-colored substance collected from the pears pericarp effected, were identified successfully for the first time, which were just rhein, aloe-emodin, chrysophanol and emodin, respectively. Taken together with microstructure characteristics for rust-spots of Kurle pear pericarp, it was no doubt that these eluates were the main factors affecting the rust spots on the pericarp of the Kurle pears during the atmosphere-controlled storage, which was a sign and consequence resisting the undesirable stress of the external environment.

plant biology↗

bioRxiv2017

Spatial Specificity Of Auxin Responses Coordinates Wood Formation

Spatial organization of signaling events of the phytohormone auxin is fundamental for maintaining a dynamic transition from plant stem cells to differentiated descendants. The cambium, the stem cell niche mediating wood formation, fundamentally depends on auxin signaling but its exact role and spatial organization is obscure. Here, we show that, while auxin signaling levels increase in differentiating cambium descendants, a moderate level of signaling in cambial stem cells is essential for cambium activity. We identify the auxin-dependent transcription factor ARF5/MONOPTEROS to cell-autonomously restrict the number of stem cells by attenuating the activity of the stem cell promoting WOX4 gene. In contrast, ARF3 and ARF4 function as cambium activators in a redundant fashion from outside of WOX4-expressing cells. Our results reveal an influence of auxin signaling on distinct cambium features by specific signaling components and allow the conceptual integration of plant stem cell systems with distinct anatomies.

plant biology↗

bioRxiv2017

Discovery Of Drought-Responsive Transposable Element-Related Peach miRNAs

MicroRNAs (miRNA) are small non-coding regulatory RNAs that suppress their specific target transcripts either by cleavage or inhibition of translation. Transposable element related miRNAs (TE-miRNA) have been subjected to various studies so far, and some of them were found to be involved in stress response in plants. Here, small RNA (sRNA) sequencing libraries generated from drought stress treated peach tissues were utilized to identify TE-miRNAs. Our computational analysis led to the identification of 63 TE-miRNAs, which were either locating nearby of any TE (TE-related), or overlapping with a TE (TE-derived). Furthermore, 13 out of 63 TE-miRNAs were designated as drought-responsive. Expression pattern of the identified drought-responsive TE-miRNAs are observed as tissue-specific manner, and their specific target transcripts are mostly related to transcription factors and growth-associated genes. Our findings suggest that miRNAs in relation with transposable elements might be key molecular players in the regulation of drought response.

plant biology↗

bioRxiv2017

Lipid transfer from plants to arbuscular mycorrhiza fungi

Arbuscular mycorrhiza (AM) symbioses contribute to global carbon cycles as plant hosts divert up to 20% of photosynthate to the obligate biotrophic fungi. Previous studies suggested carbohydrates as the only form of carbon transferred to the fungi. However, de novo fatty acid (FA) synthesis has not been observed in AM fungi in absence of the plant. In a forward genetic approach, we identified two Lotus japonicus mutants defective in AM-specific paralogs of lipid biosynthesis genes (KASI and GPAT6). These mutants perturb fungal development and accumulation of emblematic fungal 16:1{omega}5 FAs. Using isotopolog profiling we demonstrate that 13C patterns of fungal FAs recapitulate those of wild-type hosts, indicating cross-kingdom lipid transfer from plants to fungi. This transfer of labelled FAs was not observed for the AM-specific lipid biosynthesis mutants. Thus, growth and development of beneficial AM fungi is not only fueled by sugars but depends on lipid transfer from plant hosts.

plant biology↗

bioRxiv2017

Coding sequence of firefly luciferase reporter gene affects specific hyper expression in cpl1 mutant.

Forward genetic screening of mutants using firefly luciferase (LUC) reporter gene became a standard practice in plant research. Such screenings frequently identified alleles of CPL1 (Carboxyl-terminal Phosphatase-Like 1) regardless of promoters or pathways studied. Expression of the corresponding endogenous genes often shows the minimal difference between wild type and cpl1. Here we show that the LUC coding sequence is responsible for the high expression in cpl1, using a classical RD29a-LUC. Deletion of the LUC 3-UTR did not change hyperactivation of LUC in cpl1. However, a codon-modified LUC (LUC2) produced similar expression levels both in wild type and in cpl1. These results indicate that the coding region of LUC is responsible for the cpl1-specific LUC overexpression uncoupled with the expression of the endogenous counterpart.

plant biology↗

bioRxiv2017

Alkyl Gallates Display Elicitor Activities In Tobacco Plants

Alkyl gallates showed elicitor activities on tobacco in both whole plants and cell suspensions. Methyl gallate (MG), ethyl gallate (EG) and n-propyl gallate (PG) infiltration into tobacco leaves induced hypersensitive reaction-like lesions and topical production of autofluorescent compounds revealed under UV light. When sprayed on tobacco plants at 5 mM, EG promoted upregulation of defence-related genes such as the antimicrobial PR1, {beta}-1,3-glucanase PR2, chitinase PR3 and osmotin PR5 target genes. Tobacco BY-2 cells challenged with EG underwent cell death in 48 h, significantly reduced in the presence of the protease inhibitor aprotinin. The three alkyl gallates all caused alkalinisation of the BY-2 extracellular medium, whereas gallic acid did not trigger any pH variation. Using EGTA or LaCl3, we showed that Ca2+ mobilisation occurred in BY-2 cells elicited with EG. Overall, our findings are the first evidence of alkyl gallate elicitor properties with early perception events on plasma membrane, potential hypersensitive reactions and PR-related downstream defence responses in tobacco.\n\nHighlights- Alkyl gallates elicited defence reactions in tobacco\n- Alkyl gallates induced local biochemical changes in tobacco leaves\n- Alkyl gallates caused modification of plasma membrane properties\n- Ethyl gallate led to defence transcript accumulation and dose-dependent cell death associated with hypersensitive response\n- Alkyl gallates are novel elicitor agents well-suited to crop protection schemes.

plant biology↗

bioRxiv2017

Sugar metabolism changes in response to the ultraviolet B irradiation of peach (Prunus persica L.)

The protected cultivation of peach (Prunuspersica L.) trees is more economical and efficient than traditional cultivation, resulting in increased farmers incomes, but the peach sugar contents are lower than in open planting. In the greenhouse, a high-sugar variety of peach Lumi 1 was irradiated with 1.44 KJ{middle dot}m-2.d-1 intensity ultraviolet B radiation. The soluble sugar contents in fruit, peel and leaf were quantified using liquid chromatography. Overall, sucrose and sorbitol increased before the second fruit-expansion period. To further understand the mechanisms regulating sucrose and sorbitol accumulation in peach fruit, expression profiles of genes involved in sugar metabolism and transport were measured. The activity and translocation protein contents of these enzymes were measured by enzyme-linked immunosorbent assay. The increased sucrose synthase activity and sucrose transporter level in the pericarp promoted the synthesis of sucrose and intake of sucrose into fruit. Sorbitol transport into fruit was promoted by the increased sorbitol transporter protein levels in leaves. In summary, greenhouse the sucrose and sorbitol contents were increased when supplemented with 1.44 kJ{middle dot}m-2{middle dot}d-1 ultraviolet B radiation before the second fruit-expansion period of peach.

plant biology↗

bioRxiv2017

A Century of Guayule: Comprehensive Genetic Characterization of the Guayule (Parthenium argentatum A. Gray) USDA Germplasm Collection

The fragility of a single-source, geographically concentrated supply of natural rubber, a critical material of the modern economy, has brought guayule (Parthenium argentatum A. Gray) to the forefront as an alternative source of natural rubber. The improvement of guayule for commercial-scale production has been limited by the lack of genomic tools and well-characterized genetic resources required for genomics-assisted breeding. To address this issue, we developed nearly 50,000 single nucleotide polymorphism (SNP) genetic markers and genotyped 69 accessions of guayule and its sister taxa mariola (Parthenium incanum Kunth), representing the entire available NALPGRU germplasm collection. We identified multiple interspecific hybrid accessions previously considered guayule, including six guayule-mariola hybrids and non-mariola interspecific hybrid accessions AZ-2 and AZ-3, two commonly used high-yielding cultivars. We dissected genetic diversity within the collection to identify a highly diverse subset of guayule accessions, and showed that wild guayule stands in Big Bend National Park, Texas, USA have the potential to provide hitherto untapped guayule genetic diversity. Together, these results provide the most thorough genetic characterization of guayule germplasm to date and lay the foundation for rapid genetic improvement of commercial guayule germplasm.\n\nKey Results O_LISix guayule accessions are guayule-mariola hybrids\nC_LIO_LIGuayule collections from Big Bend National Park contain novel guayule genotypes not present in collections from Mexico\nC_LIO_LICommonly cultivated accessions AZ2 and AZ3 contain introgressions from other Parthenium species\nC_LIO_LIThe triploid accessions 11591, 11646, N576, N565, N565II, and RICHARDSON are generally indistinguishable from each other with respect to genetic background and likely represent the 4265-I source genotype (Johnson, 1950)\nC_LIO_LIOpen pollinated and purposefully outcrossed tetraploid selections derived from 4265-I incorporate further genetic diversity and form distinct genotypes\nC_LI

plant biology↗

bioRxiv2017

Novel multiscale insights into the composite nature of water transport in roots

- MECHA is a novel mathematical model that computes the flow of water through the walls, membranes and plasmodesmata of each individual cell throughout complete root cross-sections, from a minimal set of cell level hydraulic properties and detailed root anatomical descriptions.\n- Using the hydraulic anatomical framework of the Zea mays root reveals that hydraulic principles at the cell and root segment scales, derived independently by Katchalsky and Curran [1967] and Fiscus and Kramer [1975], are fully compatible, irrespective of apoplastic barriers leakiness.\n- The hydraulic anatomy model accurately predicts empirical root radial permeability (kr) from relatively high cell wall hydraulic conductivity and low plasmodesmatal conductance reported in the literature.\n- MECHA brings novel insights into contradictory interpretations of experiments from the literature by quantifying the impact of intercellular spaces, cortical cell permeability and plasmodesmata among others on root kr, and suggests new experiments efficiently addressing questions of root water relations.\n\n\nSymbols

plant biology↗

bioRxiv2017

Three-dimensional plant architecture and sunlit-shaded dynamics: A stochastic model to enable high-throughput physiological analysis

Background and AimsDiurnal changes in solar position and intensity combined with the structural complexity of plant architecture result in highly variable and dynamic light patterns within the plant canopy. This affects productivity through the complex ways that photosynthesis responds to changes in light intensity. Current methods to characterise light dynamics, such as ray-tracing, are able to produce data with excellent spatio-temporal resolution but are computationally intensive and the resultant data are complex and high dimensional. This necessitates development of more economical models for summarising the data and for simulating realistic light patterns over the course of a day.\n\nMethodsHigh-resolution reconstructions of field-grown plants are assembled in various configurations to form canopies, and a forward ray-tracing algorithm is applied to the canopies to compute light dynamics at high (1 minute) temporal resolution. From the ray-tracer output, the sunlit or shaded state for each patch on the plants is determined, and these data are used to develop a novel stochastic model for the sunlit-shaded patterns. The model is designed to be straightforward to fit to data using maximum likelihood estimation, and fast to simulate from.\n\nKey ResultsFor a wide range of contrasting 3D canopies, the stochastic model is able to summarise, and replicate in simulations, key features of the light dynamics. When light patterns simulated from the stochastic model are used as input to a model of photoinhibition, the predicted reduction in carbon gain is similar to that from calculations based on the (extremely costly) ray-tracer data.\n\nConclusionsThe model provides a way to summarise highly complex data in a small number of parameters, and a cost-effective way to simulate realistic light patterns. Simulations from the model will be particularly useful for feeding into larger-scale photosynthesis models for calculating how light dynamics affects the photosynthetic productivity of canopies.

plant biology↗

bioRxiv2017

A group of nuclear factor Y transcription factors are likely sub-functionalized in the endosperm development of rice and other monocots

Nuclear factor Y (NF-Y) is a heterotrimeric transcription factor that consists of three subunits, NF-YA, NF-YB, and NF-YC. Although NF-Ys play multiple roles in plant development, their functions during endosperm development are not well understood. In this study, we identified eight NF-Y encoding genes, including OsNF-YA8, OsNF-YB1,9, and OsNF-YC8,9,10,11,12, which predominantly express in the rice endosperm. Interestingly, the closest homologs of these OsNF-Ys are present only in the monocot species. All the genes are preferentially expressed in the endosperm, suggesting their roles in the regulation of endosperm development. A systemic analysis of the interactions between rice endosperm-preferential NF-Ys in yeast revealed that NF-YBs and NF-YCs could interact with each other. OsNF-YA8 is a recently evolved NF-YA in rice. Generally, NF-YA does not interact with NF-YB monomers in plants; however, in the present study, we found that OsNY-YA8 interacts with OsNF-YB9. Our results also indicated that the endosperm-preferential OsNF-YBs and OsNF-YCs could interact with some ethylene response factors (ERFs) of rice. Unlike the OsNF-YC8,9,10, the members of OsNF-YB1, 9 or OsNF-YC 11 and 12 showed lack of transcriptional activation when present alone. However, they displayed functional activity while in dimer form. In addition, OsNF-YB1 knockout lines showed significant changes in the seed morphology, further confirms its role in endosperm development. Our findings have provided strong evidences that the group of phylogenetically conserved NF-Ys are differentiated in monocots to regulate the endosperm development.

plant biology↗

bioRxiv2017

Low Rate of Somatic Mutations in a Long-Lived Oak Tree

Because plants do not possess a proper germline, deleterious somatic mutations can be passed to gametes and a large number of cell divisions separating zygote from gamete formation in long-lived plants may lead to many mutations. We sequenced the genome of two terminal branches of a 234-year-old oak tree and found few fixed somatic single-nucleotide variants (SNVs), whose sequential appearance in the tree could be traced along nested sectors of younger branches. Our data suggest that stem cells of shoot meristems are robustly protected from accumulation of mutations in trees.

plant biology↗

bioRxiv2017

High fidelity detection of crop biomass QTL from low-cost imaging in the field

Above-ground biomass production is a key target for studies of crop abiotic stress tolerance, disease resistance and yield improvement. However, biomass is slow and laborious to evaluate in the field using traditional destructive methods. High-throughput phenotyping (HTP) is widely promoted as a potential solution that can rapidly and non-destructively assess plant traits by exploiting advances in sensor and computing technology. A key potential application of HTP is for quantitative genetics studies that identify loci where allelic variation is associated with variation in crop production. And, the value of performing such studies in the field, where environmental conditions match that of production farming, is recognized. To date, HTP of biomass productivity in field trials has largely focused on expensive and complex methods, which - even if successful - will limit their use to a subset of wealthy research institutions and companies with extensive research infrastructure and highly-trained personnel. Even with investment in ground vehicles, aerial vehicles and gantry systems ranging from thousands to millions of dollars, there are very few examples where Quantitative trait loci (QTLs) detected by HTP of biomass production in a field-grown crop are shown to match QTLs detected by direct measures of biomass traits by destructive harvest techniques. Until such proof of concept for HTP proxies is generated it is unlikely to replace existing technology and be widely adopted. Therefore, there is a need for methods that can be used to assess crop performance by small teams with limited training and at field sites that are remote or have limited infrastructure. Here we use an inexpensive and simple, miniaturized system of hemispherical imaging and light attenuation modeling to identify the same set of key QTLs for biomass production as traditional destructive harvest methods applied to a field-grown Setaria mapping population. This provides a case study of a HTP technology that can deliver results for QTL mapping without high costs or complexity.

plant biology↗

bioRxiv2017

An evolutionary young defense metabolite influences the root growth of plants via the ancient TOR signaling pathway

To optimize fitness a plant should monitor its metabolism to appropriately control growth and defense. Primary metabolism can be measured by the universally conserved TOR (Target of Rapamycin) pathway to balance growth and development with the available energy and nutrients. Recent work suggests that plants may measure defense metabolites to potentially provide a strategy ensuring fast reallocation of resources to coordinate plant growth and defense. There is little understanding of mechanisms enabling defense metabolite signaling. To identify mechanisms of defense metabolite signaling, we used glucosinolates, an important class of plant defense metabolites. We report novel signaling properties specific to one distinct glucosinolate, 3- hydroxypropyl glucosinolate across plants and fungi. This defense metabolite, or derived compounds, reversibly inhibits root growth and development. 3-hydroxypropyl glucosinolate signaling functions via genes in the ancient TOR pathway. Thus, plants might link evolutionarily new defense metabolites to ancient signaling pathways to optimize energy allocation.

plant biology↗

bioRxiv2017

Arabidopsis U1 snRNP subunit LUC7 functions in alternative splicing and preferential removal of terminal introns

Introns are removed by the spliceosome, a large macromolecular complex composed of five ribonucleoprotein subcomplexes (U snRNP). The U1 snRNP, which binds to 5 splice sites, plays an essential role in early steps of the splicing reaction. Here, we show that Arabidopsis LUC7 proteins, which are encoded by a three-member gene family in Arabidopsis, are important for plant development and stress resistance. We show that LUC7 are U1 snRNP accessory proteins by RNA immunoprecipitation experiments and LUC7 protein complex purifications. Transcriptome analyses revealed that LUC7 proteins are not only important for constitutive splicing, but also affects hundreds of alternative splicing events. Interestingly, LUC7 proteins specifically promote splicing of a subset of terminal introns. Splicing of LUC7-dependent introns is a prerequisite for nuclear export and some splicing events are modulated by stress in a LUC7-dependent manner. Taken together our results highlight the importance of the U1 snRNP component LUC7 in splicing regulation and suggest a previously unrecognized role of a U1 snRNP accessory factor in terminal intron splicing.

plant biology↗

bioRxiv2017

TET-mediated epimutagenesis of the Arabidopsis thaliana methylome

DNA methylation in the promoters of plant genes sometimes leads to transcriptional repression, and the wholesale removal of DNA methylation as seen in methyltransferase mutants results in drastic changes in gene expression and severe developmental defects. However, many cases of naturally-occurring DNA methylation variations have been reported, whereby the altered expression of differentially methylated genes is responsible for agronomically important traits. The ability to manipulate plant methylomes to generate populations of epigenetically distinct individuals could provide invaluable resources for breeding and research purposes. Here we describe \"epimutagenesis\", a novel method to rapidly generate variation of DNA methylation through random demethylation of the Arabidopsis thaliana genome. This method involves the expression of a human Ten-eleven translocation (TET) enzyme, and results in widespread hypomethylation that can be inherited to subsequent generations, mimicking mutants in the maintenance DNA methyltransferase met1. Application of TET-mediated epimutagenesis to agriculturally significant plants may result in differential expression of alleles normally silenced by DNA methylation, uncovering previously hidden phenotypic variations.

plant biology↗

bioRxiv2017

Photosynthesis is heavily chlororespiration-sensitive under fluctuating light conditions

Photosynthesis needs to adjust to dynamically changing light intensities in order to maximize its efficiency, notably by the employment of alternative electron pathways. One of them is chlororespiration - initially described in Chlamydomonas reinhardtii. This electron transfer pathway, found in all photosynthetic lineages, consists of a reduction of plastoquinone (PQ) through an NAD(P)H:PQ oxidoreductase and quinol (PQH2) oxidation by Plastid Terminal Oxidase, PTOX. Hence, chlororespiration constitutes an electron pathway potentially antagonistic to the linear photosynthetic electron flow from H2O to CO2. However, the limited flow chlororespiratory enzymes can sustain suggests that their relative contribution, at least in the light and in steady-state conditions, is insubstantial. Here, we focused on the involvement of PTOX in Chlamydomonas reinhardtii during transitions from dark to light and vice versa. We show that the kinetics of redox relaxation of the chloroplast in the dark was greatly affected when PTOX2, the major plastoquinol oxidase in Chlamydomonas, is lacking. Importantly, we show that this has a direct physiological relevance, as the growth of a PTOX2-lacking mutant is markedly slower in intermittent light. The latter can be rationalized in terms of a decreased flux sustained by photosystem II due to a redox limitation at the acceptor side of the PSI during the illumination periods. We finally show that the long-term regulation of cyclic electron flow around PSI is strongly affected in the PTOX2 mutant, substantiating an important role of chlororespiration in the maintenance of chloroplast redox balance.

plant biology↗

bioRxiv2017

Persistent homology demarcates a leaf morphospace

Current morphometric methods that comprehensively measure shape cannot compare the disparate leaf shapes found in seed plants and are sensitive to processing artifacts. We explore the use of persistent homology, a topological method applied across the scales of a function, to overcome these limitations. The described method isolates subsets of shape features and measures the spatial relationship of neighboring pixel densities in a shape. We apply the method to the analysis of 182,707 leaves, both published and unpublished, representing 141 plant families collected from 75 sites throughout the world. By measuring leaves from throughout the seed plants using persistent homology, a defined morphospace comparing all leaves is demarcated. Clear differences in shape between major phylogenetic groups are detected and estimates of leaf shape diversity within plant families are made. This approach does not only predict plant family, but also the collection site, confirming phylogenetically invariant morphological features that characterize leaves from specific locations. The application of a persistent homology method to measure leaf shape allows for a unified morphometric framework to measure plant form, including shape and branching architectures.

plant biology↗