Lignified Tissues Research Articles

Proteomic and metabolic disturbances in lignin-modified Brachypodium distachyon.

Lignin biosynthesis begins with the deamination of phenylalanine and tyrosine (Tyr) as a key branch point between primary and secondary metabolism in land plants. Here, we used a systems biology approach to investigate the global metabolic responses to lignin pathway perturbations in the model grass Brachypodium distachyon. We identified the lignin biosynthetic protein families and found that ammonia-lyases (ALs) are among the most abundant proteins in lignifying tissues in grasses. Integrated metabolomic and proteomic data support a link between lignin biosynthesis and primary metabolism mediated by the ammonia released from ALs that is recycled for the synthesis of amino acids via glutamine. RNA interference knockdown of lignin genes confirmed that the route of the canonical pathway using shikimate ester intermediates is not essential for lignin formation in Brachypodium, and there is an alternative pathway from Tyr via sinapic acid for the synthesis of syringyl lignin involving yet uncharacterized enzymatic steps. Our findings support a model in which plant ALs play a central role in coordinating the allocation of carbon for lignin synthesis and the nitrogen available for plant growth. Collectively, these data also emphasize the value of integrative multiomic analyses to advance our understanding of plant metabolism.

Processing of sunflower hulls and peat with production of melanin and composite fuel briquettes

The article discusses production of composite peat fuel briquettes from butter-making waste represented by sunflower hulls. The experiments prove manufacturability of briquettes at the quality of the standard first-grade peat fuel briquettes. The specific heats of sunflower hulls and composite briquettes are determined. Furthermore, the experimental tests show that ultrasonic treatment used to extract melanin from sunflower hulls changes the chemical composition of the raw material and its reactivity worth, as well as promotes an increase in the specific heat of sunflower hulls from 17.5 to 19.7 MJ/kg. The ultrasonic treatment degrades lignified tissue matrix of sunflower hulls. The Fouriertransform infrared spectra of lignin from sunflower hulls are analyzed before and after the ultrasonic treatment. Despite the strong resemblance, the spectra display differences in the structure of lignin, as well as in the ranges of fluctuation of methyl and methylene groups. It is shown that the yield of melanin from sunflower hulls in the optimized extraction conditions in the ultrasonic field (temperature 60 0С, extraction time 25 min, density 414 W/cm2, sunflower hulls : extractant ratio 1 : 40) totals 7.6% of the biomass of sunflower hulls. The size of melanin particles is 457–465 nm. The accomplished research proves feasibility of the integrated approach to processing of sunflower hulls to produce bio-active melanin and composite peat fuel briquettes.The study was supported by the Russian Science Foundation, Project No. 20-69-47084.

Plant hormones coordinate monolignol biosynthesis with seasonal changes in Populus tomentosa

Temperate woody plants suffer environmental stresses caused by seasonal changes. Seasonal changes affect tree cambium activity, resulting in the annual periodicity of growth. In the present study, we provide a model for plant hormone coordination of monolignol biosynthesis to cope with the seasonal changes in a poplar species. First, 27 core lignin-related genes were identified in Populus tomentosa , and co-expression in lignifying tissues was identified. Combining spatial and seasonal expression analysis and the seasonal dynamic changes in plant hormones of Populus tomentosa , we found that poplars reorganized plant hormone (cytokinin；3-indoleacetic acid, IAA; and abscisic acid, ABA) biosynthesis and transport pathways with a winter-down pattern, resulting in decreases of cytokinin, IAA, and ABA content in cambium cells. The same expression pattern was also found for lignin-related genes involved in monolignol precursors biosynthesis. The monolignol specific pathway genes PtoCAD1 , 2 , 8 , PtoCCR1 , 7 and PtoF5H2 showed contrasting winter-up expression patterns. We propose that 1) plant hormones， such as cytokinin, IAA, and ABA regulate the cambium activity and coordinate monolignol biosynthesis, especially the components upstream of this pathway, with seasonal changes in Populus tomentosa ; 2) the inhibition of the biosynthesis of hydroxycinnamoyl-CoA esters and the accumulation of monolignol in winter are responsible for surviving the cold winter and provide materials for fast growth during the following spring. • Seasonal changes affect tree cambium activity and temperate woody plants growth. • 27 lignin-related genes fell into two different expression clusters in poplar. • Plant hormone related pathways were reorganized during seasonal changes. • Plant hormones coordinate cambium activity and monolignol biosynthesis with seasonal changes.

Characterization and Pathogenicity of Diplodia, Lasiodiplodia, and Neofusicoccum Species Causing Botryosphaeria Canker and Dieback of Apple Trees in Central Chile.

In recent years, the number of apple trees affected by Botryosphaeria cankers and dieback has considerably increased in central Chile. This study aimed to identify the species of Botryosphaeriaceae associated with canker and dieback symptoms, estimate disease incidence and distributions, and study their pathogenicity and virulence on apple and other fruit crops. A field survey of 34 commercial orchards of apple (7 to 30 years old) was conducted in 16 localities, obtaining 270 symptomatic branch and trunk samples in 2017 and 2018 growing seasons. The incidence of Botryosphaeria canker and dieback ranged between 5 and 40%, and a total of 255 isolates of Botryosphaeriaceae spp. were obtained from 238 cankers. Morphological identification along with phylogenetic studies of the internal transcribed spacer (ITS) region (ITS1-5.8S-ITS2) of the rDNA, part of the translation elongation factor 1-α (tef1-α), and part of the β-tubulin (tub2) genes allowed us to identify Diplodia mutila (n = 49 isolates), D. seriata (n = 136 isolates), Lasiodiplodia theobromae (n = 16 isolates), and Neofusicoccum arbuti (n = 54 isolates). L. theobromae was isolated mainly from apple dieback from northern localities. All pathogens tested were pathogenic, causing canker and dieback symptoms on lignified twigs of apple, pear, walnut, and green grapevine shoots in the field. Isolates of N. arbuti were the most virulent, reproducing more severe cankers on the lignified tissues inoculated. This study reports, for the first time, D. mutila and L. theobromae associated with Botryosphaeria canker and dieback in Chile, and it is the first description of N. arbuti causing apple dieback worldwide.

CHARACTERIZATION OF Exserohilum turcicum INFECTION SITES IN MAIZE GENOTYPES

ABSTRACT Northern leaf blight caused by Exserohilum turcicum is an important disease of maize (Zea mays L.), and its severity depends more closely on growth lesions than on spot number. Here, we characterized the infection sites of E. turcicum on resistant and susceptible maize genotypes by analyzing the histology of lesions as well as the structural and biochemical mechanisms of infection. Maize leaves were inoculated with the pathogen at specific points and incubated in a microhumidity chamber. Samples were obtained to follow fungal development and host tissue lignification using light and electron microscopy, and the activity and electrophoretic patterns of peroxidases were determined. The time course of spore germination and appressorium formation was essentially the same for both genotypes; however, a delay of 12 h in fungal penetration, accompanied by host tissue lignification, was noted in the resistant genotype, as opposed to that in the susceptible one. Scanning electron microscopy revealed fungal mycelium in the xylem vessels of both genotypes; however, in the resistant genotype, pathogen colonization was restricted to mesophyll cells around the penetration point, where chlorotic flecks were produced. Meanwhile, in the susceptible genotype, following penetration and chlorotic fleck formation, the pathogen continued to grow inside the xylem vessels and profusely colonized mesophyll tissue distant from the penetration point, resulting necrotic lesion development. Electrophoretic patterns of peroxidases were similar between the two genotypes, with three isoenzymes present in all tissues. In addition, two novel isoenzymes were detected in chlorotic flecks, necrotic lesions, and green tissue around the lesions.

Characterization and Interaction Analysis of the Secondary Cell Wall Synthesis-Related Transcription Factor PmMYB7 in Pinus massoniana Lamb.

In vascular plants, the importance of R2R3-myeloblastosis (R2R3-MYB) transcription factors (TFs) in the formation of secondary cell walls (SCWs) has long been a controversial topic due to the lack of empirical evidence of an association between TFs and downstream target genes. Here, we found that the transcription factor PmMYB7, which belongs to the R2R3-MYB subfamily, is involved in lignin biosynthesis in Pinus massoniana. PmMYB7 was highly expressed in lignified tissues and upon abiotic stress. As a bait carrier, the PmMYB7 protein had no toxicity or autoactivation in the nucleus. Forty-seven proteins were screened from the P. massoniana yeast library. These proteins were predicted to be mainly involved in resistance, abiotic stress, cell wall biosynthesis, and cell development. We found that the PmMYB7 protein interacted with caffeoyl CoA 3-O-methyltransferase-2 (PmCCoAOMT2)—which is involved in lignin biosynthesis—but not with beta-1, 2-xylosyltransferase (PmXYXT1) yeast two-hybrid (Y2H) studies. Our in vivo coimmunoprecipitation (Co-IP) assay further showed that the PmMYB7 and PmCCoAOMT2 proteins could interact. Therefore, we concluded that PmMYB7 is an upstream TF that can interact with PmCCoAOMT2 in plant cells. These findings lay a foundation for further research on the function of PmMYB7, lignin biosynthesis and molecular breeding in P. massoniana.

Genome-Wide Analysis of the Peroxidase Gene Family and Verification of Lignin Synthesis-Related Genes in Watermelon.

Watermelon (Citrullus lanatus) is an important horticultural crop worldwide, but peel cracking caused by peel hardness severely decreases its quality. Lignification is one of the important functions of class III peroxidase (PRX), and its accumulation in the plant cell wall leads to cell thickening and wood hardening. For in-depth physiological and genetical understanding, we studied the relationship between peel hardness and lignin accumulation and the role of PRXs affecting peel lignin biosynthesis using genome-wide bioinformatics analysis. The obtained results showed that lignin accumulation gradually increased to form the peel stone cell structure, and tissue lignification led to peel hardness. A total of 79 ClPRXs (class III) were identified using bioinformatics analysis, which were widely distributed on 11 chromosomes. The constructed phylogenetics indicated that ClPRXs were divided into seven groups and eleven subclasses, and gene members of each group had highly conserved intron structures. Repeated pattern analysis showed that deletion and replication events occurred during the process of ClPRX amplification. However, in the whole-protein sequence alignment analysis, high homology was not observed, although all contained four conserved functional sites. Repeated pattern analysis showed that deletion and replication events occurred during ClPRXs’ amplification process. The prediction of the promoter cis-acting element and qRT-PCR analysis in four tissues (leaf, petiole, stem, and peel) showed different expression patterns for tissue specificity, abiotic stress, and hormone response by providing a genetic basis of the ClPRX gene family involved in a variety of physiological processes in plants. To our knowledge, we for the first time report the key roles of two ClPRXs in watermelon peel lignin synthesis. In conclusion, the extensive data collected in this study can be used for additional functional analysis of ClPRXs in watermelon growth and development and hormone and abiotic stress response.

Tissue-specific transcriptomics reveals a central role of <i>CcNST1</i> in regulating the fruit lignification pattern in <i>Camellia chekiangoleosa</i>, a woody oil-crop

Fruit lignification is of significant economic importance because it affects the quality of fruit and the production of seed oil. The specified lignification pattern in<italic> Camellia</italic> <italic>chekiangoleosa</italic> fruits plays critical roles in its seed oil yield, but little is known about how this lignification process is regulated. Here, we report on a comprehensive tissue-specific transcriptomics analysis conducted for <italic>C.</italic> <italic>chekiangoleosa</italic> fruit. By mining the differentially expressed genes, we found that lignin biosynthesis and transcriptional regulation pathways were significantly enriched in the lignified tissues. The homolog of NST-like transcription factor, <italic>CcNST1</italic>, was highly expressed in lignified seed coat and endocarp tissues; transgenic analyses of <italic>CcNST1</italic> in <italic>Arabidopsis</italic> and hybrid poplar revealed the enhanced lignification levels of various tissues. Gene expression analysis of the transgenic lines uncovered potential downstream genes involved in the regulation of lignin biosynthesis. This work provides a valuable gene expression resource and identified the pivotal role of <italic>CcNST1</italic> in regulating the lignin biosynthesis underlying fruit lignification.

Dental microwear foraging ecology of a large browsing ruminant in Northern Hemisphere: The European moose (Alces alces)

Years of studies have already highlighted the complex combination, in moose feeding ecology, of a marked selectivity coupled with a significant dietary adaptability toward changes in the local resource availability. Dental textures resulting from masticatory movements and the properties of ingested food items constitute a link between the animal, its ecology and the environment it occupies. This approach is efficient to decipher subtle variations in diet, at the interspecific but also intra-populational scales. In this study, we explore inter and intra population dietary variations among six Northern European moose populations using DMTA. We show that moose feeding ecology spans a continuum between a diet dominated by tender leaves and a diet consisting of lignified tissues. The structure of habitats is the main driver of these dietary differences between populations. The absence of significant variation between males and females or between seasons is interpreted as a reflection of the food selectivity of this deer on a finer scale. The moose has a long common history with humans, constituting at certain times and in certain places the main food resource of these populations, adapting in other contexts and at other times to the repercussions of increasing anthropization and global climate change. We aim here at characterizing the dental microwear texture diversity hidden within the “browsing” dietary category. This work is also intended to be used as a reference for future paleontological or archeological investigations. We believe that it will contribute to a better understanding of the(paleo)ecology of the species and of the variations in its feeding ecology through time.

Transcriptome and metabolite profiling to identify genes associated with rhizome lignification and the function of ZoCSE in ginger (Zingiber officinale).

Ginger (Zingiber officinale Roscoe) is an important spice crop in China, and fresh ginger rhizomes are consumed as vegetable in Sichuan and Chongqing. However, tissue lignification accelerates with rhizome maturation, resulting in the loss of edible quality. To understand the molecular mechanisms of texture modification during rhizome development, we investigated lignin accumulation patterns and identified the key genes associated with lignin biosynthesis using gas chromatography-mass spectrometry (GC-MS), liquid chromatography-tandem mass spectrometry (LC-MS/MS) and RNA-sequencing (RNA-Seq). Results showed that the contents of total lignin and its precursors exhibited notable declines with tissue maturation. However, the lignin composition was remarkably modified and syringyl lignin was deposited in mature rhizomes, leading to ginger lignification. Transcriptome analysis displayed 32 lignin biosynthetic genes were dramatically downregulated with rhizome development, including caffeoylshikimate esterase (CSE ), 4-coumarate-CoA ligase , laccase , cinnamoyl-CoA reductase , cinnamyl-alcohol dehydrogenase , peroxidase and caffeic acid 3-O-methyltransferase , indicating that lignin reduction might be attributed to deficiency in intermediates or the downregulation of key biosynthetic enzymes. Furthermore, overexpressing ZoCSE in Nicotiana benthamiana L. enhanced the total lignin content, suggesting its fundamental role in lignin biosynthesis. RNA-Seq also identified candidate lignin production regulators, including hormone-related genes and NAC/MYB transcription factors (ZoNAC1 , ZoNAC4 , ZoMYB14 and ZoMYB17 ). This result provides a molecular basis for lignin accumulation in ginger.

Darkfield and Fluorescence Macrovision of a Series of Large Images to Assess Anatomical and Chemical Tissue Variability in Whole Cross-Sections of Maize Stems.

The proportion and composition of plant tissues in maize stems vary with genotype and agroclimatic factors and may impact the final biomass use. In this manuscript, we propose a quantitative histology approach without any section labelling to estimate the proportion of different tissues in maize stem sections as well as their chemical characteristics. Macroscopic imaging was chosen to observe the entire section of a stem. Darkfield illumination was retained to visualise the whole stem cellular structure. Multispectral autofluorescence images were acquired to detect cell wall phenolic compounds after UV and visible excitations. Image analysis was implemented to extract morphological features and autofluorescence pseudospectra. By assimilating the internode to a cylinder, the relative proportions of tissues in the internode were estimated from their relative areas in the sections. The approach was applied to study a series of 14 maize inbred lines. Considerable variability was revealed among the 14 inbred lines for both anatomical and chemical traits. The most discriminant morphological descriptors were the relative amount of rind and parenchyma tissues together with the density and size of the individual bundles, the area of stem and the parenchyma cell diameter. The rind, as the most lignified tissue, showed strong visible-induced fluorescence which was line-dependant. The relative amount of para-coumaric acid was associated with the UV-induced fluorescence intensity in the rind and in the parenchyma near the rind, while ferulic acid amount was significantly correlated mainly with the parenchyma near the rind. The correlation between lignin and the tissue pseudospectra showed that a global higher amount of lignin resulted in a higher level of lignin fluorescence whatever the tissues. We demonstrated here the potential of darkfield and autofluorescence imaging coupled with image analysis to quantify histology of maize stem and highlight variability between different lines.

Cell-wall fluorescence highlights the phases of xylogenesis

Summary The monitoring of xylogenesis makes it possible to follow tree growth responses to stress factors in real-time, by observing the course of wood cell division and differentiation. Proper microscopy techniques are of key importance to exactly identify the xylem cells during the different phases of differentiation. We aimed to apply epifluorescence microscopy to follow the lignification process during the different phases of xylogenesis in Mediterranean softwood and hardwood. Microcores from trees of Pinus halepensis Mill. and Arbutus unedo L. were collected at a site in southern Italy, during the period June-December. Fluorescence imaging of sections stained with a water solution of safranin and Astra blue clearly highlighted the contrast between lignified and un-lignified tissue. The proposed methodology is useful to quickly and unambiguously detect the different stages of cell differentiation, as well as the progress in the lignification process. Moreover, it proved to be easily applied to demanding wood materials, such as Mediterranean woods and can be helpful to better track stress responses and the development of anomalies during wood formation, such as intra-annual density fluctuations.

GC-MS, FTIR and Antiulcer Screening of Aqueous Seed Extract and Oil of Nigella sativa in Wistar Rats

Peptic ulcer disease (PUD) is a sore in the lining of the stomach or duodenal mucosa. This study was aimed at evaluating the antiulcer activity of the aqueous extract of Nigella sativa (black seed) and its constitutents. The pharmacognostic properties of the dry seeds were determined. The seeds were extracted using two methods digestion and hydro-distillation. The acute toxicity, phytochemical constituents and the antiulcer evaluation were performed on ethanol-induced ulcer in wistar rats using a standard method. FTIR and GC-MS analysis of the aqueous seed extract was also determined using standard methods. Nigella sativa seed shows the presence of starch grains, lignified tissues, tannins, cellulose, protein and oil globules. The aqueous extract has a high safety margin. The phytochemical studies revealed the presence of saponins, flavonoids, alkaloids, tannins, glycosides, fats and oil. The black seed aqueous and oil extract at 500 mg/kg significantly reduced the acidity, total acidity, and ulcer index, and pH of gastric content when compared with the positive control (Famotidine). The FTIR analysis identified the presence of the following functional groups chloro, ether, amine, carboxylic acid, nitriles, methylene, alcohol, while the GC-MS identified five compounds such as glycerin, n-Hexadecanoic acid, 9, 12-octadecadienoic acid-methyl ester, 9, 12-octadecadienoic acid and 9, 12-octadecadienoyl chloride. The pharmacognostic properties can act as a reliable tool for the standardization of the plant part. This study suggests that aqueous and oil extract possess antiulcer properties. Thus the aqueous and oil extract of black seed can be considered as antiulcer medication traditionally.&#x0D; Keywords: Nigella sativa, Black seed, Pharmacognostic, Phytochemical, Acute toxicity (LD50), Antiulcer.

Morpho-Anatomy of the Gynoecium and Fruit in Three Ornamental Members of Campanuloideae (Campanulaceae)

Abstract Gynoecium and fruit micromorphology and anatomy were studied using light microscopy in three species of Campanuloideae belonging to the tribes Cyanantheae ( Platycodon grandiflorus ), Wahlenbergieae ( Jasione montana ), and Campanuleae ( Adenophora liliifolia ) of the native and cultivated flora of Ukraine, to elucidate structural adaptations for fruit dehiscence. The studied species differed in flower/fruit orientation, carpel number, ovary insertion, and capsule-opening position, and the information provided a background for the discovery of common anatomical features influencing fruit dehiscence. In the studied species, the synascidiate and symplicate zones in the ovary and prominent placentae were found to be located near the mid-region of the ovary. The distinct innervation of the ovary wall and ovules was described. In P. grandiflorus , the septa and fruit wall contained a lignified parenchyma in a subepidermal position, while in J. montana and A. liliifolia , lignification was observed only in proximal portions of the septa, above the placenta ( J. montana ), or was noted as a narrow strand from the ovary base to the mid-region, called “axicorn” ( A. liliifolia ). In all the studied species, unlignified endocarpium was detected. The semi-inferior capsule of P. grandiflorus has been defined as a capsule of the Forsythia -type, with a sclerenchymatous layer in the inner zone of the mesocarpium. In J. montana and A. liliifolia , a new histogenetic type of capsule is described as Campanula -type, without lignified layers in the fruit wall and with lignified tissue in the septa, which enabled dehiscence. Our study proposed for the first time the classification of the capsules found in the studied species based on the development of openings in the superior or inferior regions of the fruits. The upright fruits of P. grandiflorus and J. montana reveal incomplete dorsiventral dehiscence in the superior region of the fruit, while dehiscence of pendent fruits of A. liliifolia is hippocrepiform-septifragal interlocular and occurs in the inferior region of the fruit at its base, as revealed previously in Campanula latifolia fruit.

Active Transport of Lignin Precursors into Membrane Vesicles from Lignifying Tissues of Bamboo.

Lignin is the second most abundant natural polymer on Earth and is a major cell wall component in vascular plants. Lignin biosynthesis has three stages: biosynthesis, transport, and polymerization of its precursors. However, there is limited knowledge on lignin precursor transport, especially in monocots. In the present study, we aimed to elucidate the transport mode of lignin monomers in the lignifying tissues of bamboo (Phyllostachys pubescens). The growth manners and lignification processes of bamboo shoots were elucidated, which enabled us to obtain the lignifying tissues reproducibly. Microsomal membrane fractions were prepared from tissues undergoing vigorous lignification to analyze the transport activities of lignin precursors in order to show the ATP-dependent transport of coniferin and p-glucocoumaryl alcohol. The transport activities for both precursors depend on vacuolar type H+-ATPase and a H+ gradient across the membrane, suggesting that the electrochemical potential is the driving force of the transport of both substrates. These findings are similar to the transport properties of these lignin precursors in the differentiating xylem of poplar and Japanese cypress. Our findings suggest that transport of coniferin and p-glucocoumaryl alcohol is mediated by secondary active transporters energized partly by the vacuolar type H+-ATPase, which is common in lignifying tissues. The loading of these lignin precursors into endomembrane compartments may contribute to lignification in vascular plants.

Lignin concentrations in phloem and outer bark are not associated with resistance to mountain pine beetle among high elevation pines.

A key component in understanding plant-insect interactions is the nature of host defenses. Research on defense traits among Pinus species has focused on specialized metabolites and axial resin ducts, but the role of lignin in defense within diverse systems is unclear. We investigated lignin levels in the outer bark and phloem of P. longaeva, P. balfouriana, and P. flexilis; tree species growing at high elevations in the western United States known to differ in susceptibility to mountain pine beetle (Dendroctonus ponderosae; MPB). Pinus longaeva and P. balfouriana are attacked by MPB less frequently than P. flexilis, and MPB brood production in P. longaeva is limited. Because greater lignification of feeding tissues has been shown to provide defense against bark beetles in related genera, such as Picea, we hypothesized that P. longaeva and P. balfouriana would have greater lignin concentrations than P. flexilis. Contrary to expectations, we found that the more MPB-susceptible P. flexilis had greater phloem lignin levels than the less susceptible P. longaeva and P. balfouriana. No differences in outer bark lignin levels among the species were found. We conclude that lignification in Pinus phloem and outer bark is likely not adaptive as a physical defense against MPB.

Laccase Directed Lignification Is One of the Major Processes Associated With the Defense Response Against Pythium ultimum Infection in Apple Roots.

Apple replant disease (ARD), incited by a pathogen complex including Pythium ultimum, causes stunted growth or death of newly planted trees at replant sites. Development and deployment of resistant or tolerant rootstocks offers a cost-effective, ecologically friendly, and durable approach for ARD management. Maximized exploitation of natural resistance requires integrated efforts to identify key regulatory mechanisms underlying resistance traits in apple. In this study, miRNA profiling and degradome sequencing identified major miRNA pathways and candidate genes using six apple rootstock genotypes with contrasting phenotypes to P. ultimum infection. The comprehensive RNA-seq dataset offered an expansive view of post-transcriptional regulation of apple root defense activation in response to infection from P. ultimum. Several pairs of miRNA families and their corresponding targets were identified for their roles in defense response in apple roots, including miR397-laccase, miR398-superoxide dismutase, miR10986-polyphenol oxidase, miR482-resistance genes, and miR160-auxin response factor. Of these families, the genotype-specific expression patterns of miR397 indicated its fundamental role in developing defense response patterns to P. ultimum infection. Combined with other identified copper proteins, the importance of cellular fortification, such as lignification of root tissues by the action of laccase, may critically contribute to genotype-specific resistance traits. Our findings suggest that quick and enhanced lignification of apple roots may significantly impede pathogen penetration and minimize the disruption of effective defense activation in roots of resistant genotypes. The identified target miRNA species and target genes consist of a valuable resource for subsequent functional analysis of their roles during interaction between apple roots and P. ultimum.

Green light promotes healing and root regeneration in double-root-cutting grafted tomato seedlings

Double-root-cutting (DRC) grafting is a new grafting method that makes it possible to ship grafted seedlings without using plug trays, through cutting off the roots of both scion and rootstock. However, graft healing and root regeneration are the two important factors determining the survival of the DRC grafted plants. Recent studies have shown that green light plays a crucial role in plant growth and development. We hypothesized that green light could stimulate the graft healing and root regeneration of the rootstock in DRC grafted seedlings. To test this hypothesis, we used tomato seedlings (Solanum lycopersicum cv. Beaded Curtain grafted on cv. Rootstock No. 1) exposed to white LED lamp (W) as a control (CK). In the treatment group, green light (10, 20 and 30%) was added to the red and blue spectrum without changing the intensity of red and blue fractions. The effects of different light qualities on graft union formation, survival rate, rooting of the rootstock hypocotyls, activities of antioxidant enzymes, chlorophyll content and grafted seedling quality of grafted tomato seedlings were then studied. Compared with DRC grafted seedlings exposed to the same light intensity, addition of 30% green light to red and blue lights increased the activities of antioxidant enzymes, chlorophyll content, plant height and index of seedling strength of grafted seedlings, showed a more positive effect on the development and performance of grafted seedlings, and promoted graft union, rootstock rooting and grafted seedling growth. Increasing the proportion of green light brought about better graft union and rootstock rooting. Our results showed that green light may reduce the degree of stomatal opening and transpiration rate during the early stages of grafting. Green light increased the activity of antioxidant enzymes, which was enhanced tissue lignification. Photosynthesis was driven by green light, which improved the development of aboveground parts, enhanced the transport of auxin from leaves to rooting zone, and promoted hypocotyl rooting of DRC grafted tomato seedlings.

Pharmacognostical and Phytochemical Analysis of Rasayana Churna Vati for Quality Control - An Ayurveda Formulation

Rasayana churna was mentioned by Acharya Vagbhatta for rejuvenation of body tissue. Rasayana properties of this combination revitalize the unhealthy tissue on systemic level. Rasayana Churna is a polyherbal formulation contacting 3 ingredients i.e., Amalaki, Guduchi and Gokshura are well known drugs for its rich in antioxidant, immunomodulator and purifactory action. Repeated stress injury in visual system is major cause of Computer Vision Syndrome- a new milliner disorder. A study conducted as randomized clinical trial showed that Rasayan churana significantly improves a group of symptoms in Computer Vision Syndrome.&#x0D; The present study was carried out to standardize the finished product of Rasayana Churna Vati. It includes Authentication, Organoleptic properties, physic-chemical parameters, assay of active and qualitative HPTLC analysis to confirm entity, quality and purity. All results were compared with Ayurvedic Pharmacopoeial standard. The presence of rosette crystals of calcium oxalate, lignified tissues, prismatic crystals, reticulate vessels, starch grains oval, tannins, trichomes were the characteristic features observed in the microscopy of the prepared drug. Phyto-chemical analysis showed Loss on drying 1.87% w/w, Total ash 4.35% w/w, water soluble extract 34.73% w/w, Acid insoluble ash 0.73% w/w, Alcohol soluble extractive 14.34% w/w &amp; pH 3.4. Identification of constituents of Rasayana Churna Vati was done by HPTLC scanning at 254 and 366mm gives positive test for Guduchi, Gokshura and Amalaki.

Modified staining protocol with Safranin O and Astra Blue for the plant histology

Many staining protocols are widely applied in botanical microtechniques and serve specific histological purposes. In particular, some dyes are used simultaneously to receive contrasting colorations of different chemical structures, e.g., lignin and cellulose. One of the most popular differential staining protocols is based on the Safranin O / Astra Blue dyes combination. Safranin O is a water-soluble basic dye that stains lignin in red. Astra Blue is also a water-soluble dye but having an acidic reaction, which stains cellulose in blue. Usually, a 1–2 % solution of Safranin O in distilled water or 50–70 % ethanol is applied in combination with the 0.5–1 % water solution of Astra Blue to detect lignified structures and obtain contrasting pictures convenient for the light microscopy. For a long time, Astra Blue was used exclusively with water solutions, and such recommendation without additional options is indicated on producers’ web sites. However, in 2002 it was proposed to use 1 % Astra Blue solution in 95 % ethanol to identify the lignified tissues. Later, such an ethanol solution of Astra Blue was also successfully applied by other researchers for different experimental purposes.We tested the modified staining protocol with the application of both 1 % Safranin O and Astra Blue solutions in a slightly lower concentration of ethanol (70 %) on the flower buds of Gagea lutea (Liliaceae) and found it working well. We believe that such a modified protocol with the solutions of these two dyes in 70 % ethanol allows simplifying the procedure of the plant material staining due to application of the same concentrations of dissolvent and reducing the difference in solvent concentration between two following contrasting staining solutions. Such differential staining can be effectively applied for plant histology purposes, especially where there is a need to distinguish lignified structures and secretory tissues.