Bulb Formation Research Articles

Deconvolving organogenesis in space and time via spatial transcriptomics in thick tissues.

Organ development is guided by a space-time landscape that constraints cell behavior. This landscape is challenging to characterize for the hair follicle - the most abundant mini organ - due to its complex microscopic structure and asynchronous development. We developed 3DEEP, a tissue clearing and spatial transcriptomic strategy for characterizing tissue blocks up to 400 µm in thickness. We captured 371 hair follicles at different stages of organogenesis in 1 mm 3 of skin of a 12-hour-old mouse with 6 million transcripts from 81 genes. From this single time point, we deconvoluted follicles by age based on whole-organ molecular pseudotimes to animate a stop-motion 3D atlas of follicle development along its trajectory. We defined molecular stages for hair follicle organogenesis and characterized the order of emergence for its structures, differential signaling dynamics at its top and bottom, morphogen shifts preceding and accompanying structural changes, and series of structural changes leading to the formation of its canal and opening. We further found that hair follicle stem cells and their niche are established and stratified early in organogenesis, before the formation of the hair bulb. Overall, this work demonstrates the power of increased depth of spatial transcriptomics to provide a four-dimensional analysis of organogenesis.

Photoperiod and temperature as dominant environmental drivers triggering chemical compositions of garlic bulb during growth

Photoperiod and temperature are crucial for plant growth and development, but the accumulation patterns and regulatory mechanisms of garlic in response to these factors remain unclear. Compared to normal conditions (12/12 h light/dark; 22/15 °C day/night), increasing the photoperiod by 1 h each month with normal temperature (13/11, 14/10, 15/9, 16/8 h; light/dark; 22/15 °C day/night) or increasing photoperiod by 1 h with a 1.5 °C increase in temperature (13/11, 14/10, 15/9, 16/8 h light/dark; 23.5/16, 25/17, 26.5/18, 28/19 °C day/night) at monthly intervals substantially affects garlic growth in early G204, mid-G666, and late G390 harvest cultivars. For both increased photoperiod and temperature, the environment most favorable for bulb growth is characterized by the maximum accumulation of hormones (IAA, JA, and ZT) during bulb formation, while higher levels of ACC, phenolic acids, and organosulfur compounds are observed during the harvesting stage. Furthermore, the accumulation patterns of alliin and allicin in garlic bulbs for G204, G666, and G390 cultivars significantly increased, with fold changes of (2.0, 1.8, and 1.9) and (1.8, 1.7, and 1.5), respectively, in response to increased photoperiod and temperature conditions. Additionally, the accumulation pattern in G204, and G666 was similar but notably distinct from late harvest G390 during growth. In this study, metabolomics models were constructed to assess chemical composition variations and predict the differences among three garlic varieties during the growth stages. These models revealed functional component variations and provided reliable predictions, offering valuable insights for estimating garlic chemical compositions and identifying optimal environmental conditions for growth.

Vegetative reproduction and yield of bear garlic (<i>Allium ursinum</i> L.) in field cultivation

Bear garlic (<i>Allium ursinum</i> L.) and its value for human health have been known for centuries. Its leaves, flowers, and bulbs contain many health-promoting compounds, and can be essential raw materials for the production of food and dietary supplements of natural origin. The purpose of the study was to evaluate the possibility of growing bear garlic in field conditions that differed significantly from those required by this plant, using unconventional treatments: catch crops (phacelia), shade plant (turnip rape), and biopreparations (Bio-Algeen S90 and Kelpak SL). The plowing of phacelia biomass had a beneficial effect on the number of <i>A. ursinum</i> progeny bulbs, the unit weight of bulbs and their yield per unit area, and the leaf yield not only in the first year of cultivation but also in the follow-up. Winter turnip rape applied as a shade crop caused a slight increase in the unit weight of bulbs and their yield in the first and second years of cultivation and in the number of progeny bulbs in the second and third years of cultivation. The cultivation of <i>A. ursinum</i> with turnip rape increased the percentage of garlic root weight and leaf yield. There was a positive effect of the applied biopreparations on the studied traits of <i>A. ursinum</i> bulb yield and leaf yield and on the formation of progeny bulbs in relation to the control object. The analysis of the interaction of the factors of the experiment showed that the combination with the application of the phacelia catch crop and the simultaneous cultivation of the shade crop had the most favorable effect on <i>A. ursinum</i> progeny bulb formation, bulb unit weight, root system development, and leaf yield. In all years of the study, the worst effect was obtained in the control object, i.e., in the cultivation without the catch crop, rapeseed, and application of biopreparations.

Impact of Organic Manures and Liquid Formulations on Garlic Growth, Yield and Quality in Uttarakhand, India

The field study was carried out to VCSG, UUHF, Pauri Garhwal in 2022-2023 to investigate the integrated effect of organic manures and organic liquid formulations on the growth and yield of garlic. The main objective of this study was to determine the benefits of using organic manures and liquid formulations over conventional farming because the abundant use of nitrogenous fertilizers causes excessive vegetative growth at the expense of bulb formation. Among fourteen different treatment combinations that consist of various combinations of organic manures (FYM and Vermicompost) and organic liquid formulations (Panchgavya and Jeevamrit) treatment T3 i.e., FYM (10t/ha) + Panchgavya (2%) emerged as the best treatment concerning growth and yield of garlic. The findings showed that the maximum plant height (69.080 cm), number of leaves (8.400), leaf length (47.880 cm) and breadth (3.207 cm), and leaf area index (1.023) at all the 30 days intervals after sowing of garlic has been observed with the application of Treatment T3. The fundamental reason for the increased growth might be due to growth-promoting substances like auxins and gibberellins in panchgavya and high nitrogen and phosphorus in FYM. Concerning yield and quality attributes, the bulb diameter (5.657 cm), fresh weight of bulb (64.643 cm), length of the bulb (5.853 cm), number of cloves per bulb (12.200), bulb yield (120.833 q), total chlorophyll content (2.127 mg/g) and ascorbic acid content (15.333 mg/100g) was obtained maximum with the application of same treatment T3 i.e., FYM (10t/ha) + Panchgavya (2%) with yield increases up to 30% compared to control. However, the harvest index percent was found to be maximum (89.243 %) in treatment T13 i.e., Vermicompost (5t/ha) + Jeevamrit (6%). These findings suggest that organic farming practices can effectively enhance garlic production, offering a sustainable alternative to chemical fertilizers. Moreover, this study provides practical insights for farmers and contributes to the growing body of research on sustainable agriculture.

Identification and validation of reference genes for gene expression study using Quantitative Real-Time PCR in shallot (Allium cepa L. Aggregatum group) based on bulb development phase

Abstract. Sulistyaningsih E, Sawitri WD, Handayani VDS, Wicaksono AW, Silalahi CM, Murti RH. 2024. Identification and validation of reference genes for gene expression study using Quantitative Real-Time PCR in shallot (Allium cepa L. Aggregatum group) based on bulb development phase. Biodiversitas 25: 2645-2651. Shallot (Allium cepa L. Aggregatum group) is cultivated vegetatively using bulbs (produce many bulbs) and generatively using TSS (True Seed of Shallot) which tends to produce single bulbs. Bulb aggregation is influenced by activity of genes and the gene expression could be analyzed using quantitative Real-Time Polymerase Chain Reaction (qRT-PCR). The qRT-PCR analysis requires reference gene data (housekeeping gene) as a comparison gene against our target gene. This study aimed to select and validate three housekeeping genes in shallot as an initial study for qRT-PCR analysis. The research flow began with the preparation of shallot 'TukTuk' leaves and bulb samples in bulb formation from bulb initial phase to mature phase. Furthermore, a specific primer design based on actin, tubulin, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene sequences were obtained from gene bank data. The results showed that the actin, tubulin, and GAPDH genes were successfully amplified and the products were 142 bp, 116 bp, and 136 bp in size, respectively and the genes were expressed almost constantly in leaves and bulbs in all phases of bulb formation. The tubulin gene had the potential as a reference gene for qRT-PCR analysis in shallot with the most consistently expressed.

Identification and Expression Analysis of Putative Sugar Transporter Gene Family during Bulb Formation in Lilies.

Sugar transporters play important roles in plant growth and development, flowering and fruiting, as well as responses to adverse abiotic and biotic environmental conditions. Lilies (Lilium spp.) are some of the most representative ornamental bulbous flowers. Sugar metabolism is critical for bulb formation in lilies; therefore, clarifying the amount and expression pattern of sugar transporters is essential for further analyzing their roles in bulb formation. In this study, based on the transcriptome data of the Lilium Oriental hybrid 'Sorbonne' and Lilium × formolongi, a total of 69 and 41 sugar transporters were identified in 'Sorbonne' and Lilium × formolongi, respectively, by performing bioinformatics analysis. Through phylogenetic analysis, monosaccharide transporters (MSTs) can be divided into seven subfamilies, sucrose transporters (SUTs) can be divided into three subgroups, and sugars will eventually be exported transporters (SWEETs) can be divided into four clades. According to an analysis of conserved motifs, 20, 14, and 12 conserved motifs were predicted in MSTs, SUTs, and SWEETs, respectively. A conserved domain analysis showed that MSTs and SUTs contained a single domain, whereas most of the SWEETs harbored two MtN3/saliva domains, also known as a PQ-loop repeat. The LohINT1, which was predicted to have a smaller number of transmembrane structural domains, was cloned and analyzed for subcellular localization. It was found that the LohINT1 protein is mainly localized in the cell membrane. In addition, the expression analysis indicated that 22 LohMSTs, 1 LohSUTs, and 5 LohSWEETs were upregulated in 'Sorbonne' 1 day after scale detachment treatment, suggesting that they may regulate the initiation of the bulblet. A total of 10 LflMSTs, 1 LflSUTs, and 6 LflSWEETs were upregulated 4~6 months after sowing, which corresponds to the juvenile-to-adult transition phase of Lilium × formolongi, suggesting that they may also play a role in the accompanying bulb swelling process. Combined with quantitative real-time PCR (qRT-PCR) analysis, LohSTP8 and LohSTP12 were significantly overexpressed during the extremely early stage of bulblet initiation, and LflERD6.3 was significantly overexpressed during the growth of the underground bulblet, suggesting that they may be key sugar transporters in the formation of lily bulbs, which needs further functional verification.

Effects of Priming on Onion Seed Germination and Field Performance during Summer Sowing

Aims: The aim of this study was to determine the suitable to priming treatments on seed germination, bulb production and quality of onion.
 Study Design: The experiment was set up in randomized complete block design (RCBD) with three replications.
 Place and Duration of the Study: The experiment was conducted in Horticulture farm at Sher-e Bangla Agricultural University, Dhaka, Bangladesh from April to October 2021.
 Methodology: Seeds were immersed in each priming media for 12 and 24 hours, including distilled water, IAA (100 ppm), KCl (2.5%), and PEG-6000 (10%). The treatments were consisted of T1 (water 12 hr)), T2 (water 24 hr), T3 (PEG 12 hr), T4 (PEG 24 hr), T5 (IAA 12 hr), T6 (IAA 24 hr), T7 (KCl 12 hr) and T8 (KCl 24 hr).
 Results: The maximum germination percentage (93%), plant height (25.93 cm), total chlorophyll (28.47 mg/g) of onion leaves, bulb yield (19.96 t/ha), total soluble solid (14.05 °Brix), reducing sugar (7.41%), and phenolic content (11.12 mg/100 g) of onion bulbs were recorded in the T3 treatment. However, the maximum fresh leaves/plant at bulb formation stage (3.73) and ascorbic acid content of onion bulb (7.20 mg/100 g) was recorded in T4 treatment.
 Conclusion: It can be concluded that seed priming with PEG6000 (10%) for 12 hours improved growth, yield, and quality compared to other priming treatments.

Comparative effectiveness of the use of mineral and organic fertilizers in the cultivation of onions in an annual crop

We conducted a comparative study of the effect of mineral and organic fertilizers on the yield and quality of 5 medium-ripened domestic onion varieties in an annual crop on the alluvial meadow soil of the Moscow region. Laboratory and field experiments were carried out by the Department of Agriculture and Agrochemistry of the VNIIO branch of the FNCO in 2020–2022 on sufficiently cultivated medium-loamy alluvial meadow soils of the central part of the Moskvoretsky floodplain. In the experiment with the 5th varieties and the 3rd variants of the mineral fertilizer system, the commercial yield of onions was 53.0 t/ha, whereas with the use of an organic fertilizer system – 58.4 t/ha. Accordingly, the increase in yield relative to the control without fertilizers in the first case was equal to 106, in the second – 117%, marketability – 93 and 95%. It has been established that when determining the need of plants for nutrients during the growing season, it is preferable to use methods of leaf diagnostics. On average, for 5 varieties of onion, the yield increase, according to the results of plant diagnostics, turned out to be 13% more than in variants with NPK doses calculated according to soil diagnostics. Biochemical quality indicators for all onion varieties and all fertilizer variants did not differ statistically significantly from the control. The quality of onion products practically did not depend on the applied fertilizer systems. The organic fertilizer system provided the best supply of nutrients to onions in the phase of the beginning of bulb formation, which was crucial for obtaining the greatest yield in an annual crop. On average, for 3 variants of the organic system, the soil contained 2.34 mg/100 g of mineral nitrogen in the phase of the beginning of bulb formation, whereas with the use of the mineral system – 1.65 mg/100 g, or 42% less.

Thaw bulb formation surrounding warm-oil pipelines and evaluation of the cooling performance of a new air convection pipeline embankment structure

The first China-Russia crude oil pipeline (CRCOP-I) and the second China-Russia crude oil pipeline (CRCOP-II) went into operation in January 2011 and January 2018, respectively. On-site monitoring data have indicated that the seasonal thawing depth (STD) of a borehole 2 m away from CRCOP-I reached 10.8 m by the end of 2021. To improve mitigation measures of thaw settlement, an inverted T-shaped crushed-rock pipeline embankment (ITCPE) is proposed in this study. The long-term thermal activity of three types of pipeline embankments is predicted and compared with the underground structure. This study finds that none of the current pipeline structures has solved the thaw settlement issue for the CRCOP, whether with insulation layers, in an unprotected pipeline embankment (UPE), or in a traditional horizontal crushed-rock pipeline embankment (THCPE). However, the cross-ventilation attained in the proposed approach did mitigate thawing. It is estimated that after 50 years of operation, the natural permafrost table will be 2.9 m deep, whereas the proposed ITCPE central pipeline embankment could maintain the artificial permafrost table at a depth of 0.7 m. This study provides technical support for the CRCOP and provides a reference for the design and maintenance of other pipeline projects in permafrost areas worldwide.

Efecto de la pendiente del terreno y tuberías terciarias en la uniformidad de riego, bulbo húmedo y rendimiento del cultivo de arveja (<i>Pisum sativum</i>)

The Sierra of Perú region has agricultural soils with pronounced slopes that are not used throughout the year, because there are limitations when applying traditional gravity irrigation that leads to low infiltration speed, soil erosion, runoff formation, leaching, leaching of applied fertilizers and low irrigation efficiency; In that understanding, the objective of the investigation was to evaluate the effect of the slope of land and the arrangement of tertiary pipes on the uniformity of irrigation, wet bulb formation and the crop yield of peas. Three plots were conditioned with slopes of 10%, 20%and 40%; And in each plot three irrigation subunits were installed, inserting 0, 2 and 6 valves in the tertiary pipes, having a total of 09 treatments. The subunits were installed with irrigation tapes of 1.6 l/h of emitter flow at 20cm between drippers and 60cm between sides. The irrigation uniformity coefficient (CU) was determined with the Keller and Karmeli method; Treatment 7 (slope of land 40% + tertiary pipe with 06 valves) presented greater efficiency Cu = 96.30%; valued with an excellent qualifier to irrigation uniformity. All treatments have similarity in the formation of the humid bulb at 45min of irrigation; With a wet soil radius ≥10cm, depth ≥30cm on sandy frank soil. Regarding performance, treatment 3 (slope of land 10% + tertiary pipe with 02 valves) presented 10.75 t/ha.

Variation and correlation between the parameters of the bulbs of the Oriental hyacinth and the amount of precipitation

The area of the Oriental hyacinth is located in the Mediterranean dry subtropical climate zone, with dry summers and rainy winters. The Mediterranean region is characterized by an average monthly temperature of 20-25 °C in summer, 5-12 °C in winter, and an average rainfall of 400-600 mm per year. A long period of drought contributed to the formation of a special life form in the oriental hyacinth – bulbous geophyte. Despite growing in a warm subtropical climate, hyacinth belongs to cold-resistant ephemeroids, which allows it to undergo primary introduction in the temperate continental climate of the Republic of Mari El. The main method of reproduction of hyacinth, within its natural range, is vegetative division, with the formation of daughter bulbs. In natural habitat, the coefficient of vegetative reproduction is influenced by air temperature and precipitation during the active growing season. The purpose of the work was to identify the dependence of the productivity of the Oriental hyacinth on the amount of precipitation in the conditions of the Republic of Mari El. Work on the identification of dependencies was carried out in the period 2012-2015 in the open ground of the botanical garden-Institute of the State Technical University of Yoshkar-Ola. During the work, biometric parameters such as the diameter and height of the bulb, as well as the number of bulbs in one nest, were taken into account. As a result of the work carried out, it was revealed that: the level of individual variability in bulb diameter in most varieties of oriental hyacinth is included in the norm of variation, this indicates the suitability of environmental factors of the region of introduction for this parameter; the number of daughter bulbs in the nest is characterized by a high level of variability and depends on environmental conditions; the reproduction coefficient of hyacinth varieties is strongly influenced such an environmental factor as the amount of precipitation during the growing season (66.7% – 73.7%). The identification of dependencies between the amount of precipitation and the productivity of vegetative reproduction, in the conditions of the Republic of Mari El, will allow us to assess the effectiveness of the primary introduction of Oriental hyacinth. The obtained data can be used in the development of agricultural techniques for growing Oriental hyacinth in conditions of introduction.

THU580 A Subset Of Migrating Cells Co-express Gonadotropin Releasing Hormone And Prokineticin Receptor 2 During Embryonic Development

Abstract Disclosure: A.E. Danku: None. C.F. Elias: None. A subset of migrating cells co-express gonadotropin releasing hormone and prokineticin receptor 2 during embryonic development Antoinette Danku and Carol F Elias. Neuroscience Graduate Program and Department of Molecular & Integrative Physiology, University to Michigan. Reproductive vitality in humans is strongly influenced by developmental foundations. The successful migration of GnRH cells and the pulsatile secretion of gonadotropin releasing hormone (GnRH) drives pubertal onset and dictates long-term fertility outcomes. During development, GnRH neurons migrate from the nasal placode to the basal forebrain to reach their destination in the medial preoptic area (MPOA). Aberrant GnRH migration causes redirection of axonal projections to atypical brain regions and dysregulated GnRH secretion. Disruption of GnRH neuronal function causes infertility or reproductive dysfunction. These etiologies are grouped as isolated gonadotropin deficiencies (IGD). Kallmann’s syndrome (KS) is a pediatric IGD that manifests as anosmia and hormonal dysfunction that can result in pubertal delay or infertility. Children with KS often have mutations in prokineticin 2 (Prok2) or the cognate receptor (ProkR2).Prok2 and ProkR2 are required for the formation of the olfactory bulb in rodents and humans. Loss-of-function mutations in these genes also disrupts GnRH cell migration, but whether the activation of ProkR2 regulates the guidance of GnRH neurons to their final destination has not been defined. Previous studies suggested that GnRH and ProkR2 are expressed in distinct cells, but how the prokineticin system may alter GnRH migration is largely unknown. To gain insights into the underlying mechanisms associated with ProkR2-GnRH interaction and migration, we conducted single nuclei RNA sequencing in nasal-forebrain junction of embryos at embryonic day 16 (E16) and fluorescent in situ hybridization in three developmental time points: E14, E16, and E18 for validation and spatial evaluation. Using these methods, we were able to isolate GnRH and ProkR2 cells and sort their cell types into clusters. While these cells differ in their expression profiles, they both express genes associated with migration at E16. Our findings suggest that ProkR2 cells in the nasal-forebrain junction migrate along with GnRH cells. Additionally, a subset of these migrating cells co-express ProkR2 and GnRH mRNA. Further studies are needed to determine whether this subset of cells take on specialized roles in GnRH cell migration and maturation. Presentation: Thursday, June 15, 2023

High jugular bulb in patients with non-thrombotic internal jugular venous and transverse sinus stenosis: Clues to pathogenesis.

Conventional theories for jugular bulb (JB) formation are insufficient to explain the high proportion of high JB in adult patients. We aimed to study features of high JB in patients with non-thrombotic internal jugular venous stenosis (IJVS) and/or transverse sinus stenosis (TSS) to explore the pathogenesis of high JB formation. We retrospectively enrolled consecutive patients with the diagnosis of non-thrombotic IJVS and/or TSS. The relationship between IJVS and/or TSS and high JB was explored. Logistic regression analysis was performed to identify potential independent risk factors for high JB. A total of 228 patients were included in the final analyses. The proportions of IJVS, dominant-side IJVS, and non-TSS in dominant-side high JB subgroup were higher than those in nondominant-side high JB subgroup (83.3% vs. 62.5%, p < 0.001; 72.2% vs. 18.3%, p < 0.001; 43.5% vs. 29.2%, p = 0.02). Heights of JBs on dominant sides in IJVS subgroup and non-TSS subgroup were higher than those in non-IJVS subgroup and TSS subgroup (12.93 ± 2.57 mm vs. 11.21 ± 2.76 mm, p < 0.001; 12.66 ± 2.71 mm vs. 11.34 ± 2.73 mm, p = 0.003). Multivariate logistic regression indicated an independent association between dominant-side IJVS and dominant-side high JB (odds ratio, 29.40; 95% confidence interval, 11.04-78.30; p < 0.001). IJVS and asymmetric transverse sinus were independently and positively associated with high JB, especially dominant-side IJVS with dominant-side high JB, indicating a potential hemodynamic relationship between IJVS and high JB formation. Conversely, TTS might impede high JB formation.

Myelin protein zero mutation-related hereditary neuropathies: Neuropathological insight from a new nerve biopsy cohort.

Myelin protein zero (MPZ/P0) is a major structural protein of peripheral nerve myelin. Disease-associated variants in the MPZ gene cause a wide phenotypic spectrum of inherited peripheral neuropathies. Previous nerve biopsy studies showed evidence for subtype-specific morphological features. Here, we aimed at enhancing the understanding of these subtype-specific features and pathophysiological aspects of MPZ neuropathies. We examined archival material from two Central European centers and systematically determined genetic, clinical, and neuropathological features of 21 patients with MPZ mutations compared to 16 controls. Cases were grouped based on nerve conduction data into congenital hypomyelinating neuropathy (CHN; n = 2), demyelinating Charcot-Marie-Tooth (CMT type 1; n = 11), intermediate (CMTi; n = 3), and axonal CMT (type 2; n = 5). Six cases had combined muscle and nerve biopsies and one underwent autopsy. We detected four MPZ gene variants not previously described in patients with neuropathy. Light and electron microscopy of nerve biopsies confirmed fewer myelinated fibers, more onion bulbs and reduced regeneration in demyelinating CMT1 compared to CMT2/CMTi. In addition, we observed significantly more denervated Schwann cells, more collagen pockets, fewer unmyelinated axons per Schwann cell unit and a higher density of Schwann cell nuclei in CMT1 compared to CMT2/CMTi. CHN was characterized by basal lamina onion bulb formation, a further increase in Schwann cell density and hypomyelination. Most late onset axonal neuropathy patients showed microangiopathy. In the autopsy case, we observed prominent neuromatous hyperinnervation of the spinal meninges. In four of the six muscle biopsies, we found marked structural mitochondrial abnormalities. These results show that MPZ alterations not only affect myelinated nerve fibers, leading to either primarily demyelinating or axonal changes, but also affect non-myelinated nerve fibers. The autopsy case offers insight into spinal nerve root pathology in MPZ neuropathy. Finally, our data suggest a peculiar association of MPZ mutations with mitochondrial alterations in muscle.

Indirect organogenesis and in vitro bulb formation of Pancratium maritimum

Indirect organogenesis and in vitro bulb formation of Pancratium maritimum

Elicitation and Enhancement of Phenolics Synthesis with Zinc Oxide Nanoparticles and LED Light in Lilium candidum L. Cultures In Vitro

In this study, we identified and determined the content of phenolic compounds in Lilium candidum adventitious bulbs formed in vitro. HPLC analysis revealed the presence of four phenolic acids: chlorogenic, caffeic, p-coumaric, and ferulic acid. Phenolic acid content was assessed in adventitious bulbs formed in vitro on media supplemented with zinc oxide nanoparticles (ZnO NPs at 25, 50, and 75 mg/L) under fluorescent light (FL) or in darkness (D). The second experiment analyzed the effects of light-emitting diodes (LEDs) of variable light spectra on the formation of adventitious bulbs and their contents of phenolic acids. Spectral compositions of red (R; 100%), blue (B; 100%), red and blue (RB; 70% and 30%, respectively), a mix of RB and green (RBG) in equal proportions (50%), and white light (WLED, 33.3% warm, neutral, and cool light, proportionately) were used in the study. FL and D conditions were used as controls for light spectra. Bulbs grown in soil served as control samples. The most abundant phenolic acid was p-coumaric acid. Treatment with LED light spectra, i.e., RB, RBG, WLED, and B, translated into the highest p-coumaric acid concentration as compared with other treatments. Moreover, all the bulbs formed in light, including those grown on the media supplemented with ZnO NPs and under FL light, contained more p-coumaric acid than the bulbscales of the control bulbs grown in soil. On the other hand, control bulbs grown in soil accumulated about two to three times higher amounts of chlorogenic acid than those formed in vitro. We also found that the levels of all examined phenolics decreased under FL, R, and D conditions, while the bulblets formed in vitro under RB light showed the highest phenolic content. The use of ZnO NPs increased the content of p-coumaric, chlorogenic, and caffeic acid in the bulblets formed under FL as compared with those grown in darkness.

Mechanical Response of a Buried Pipeline to Permafrost Thawing Based on Sequential Coupling Method

Thawing permafrost has affected the structural integrity of buried warm pipelines in cold regions and poses an ongoing threat in the context of climate change. Therefore, characterizing variation in the engineering properties of pipeline foundation permafrost and its effect on the mechanical behavior of pipeline is important. In this paper, the ground temperature distributions around a buried warm pipeline and mechanical response of the pipeline to differential thaw settlement of foundation permafrost are investigated using thermal–mechanical sequential coupling simulation, based on the observational data collected from a selected monitoring site along the China-Russia crude oil pipelines in northeastern China. The results indicate that the thaw-induced settlement of pipeline foundation permafrost develops quickly with the formation and expansion of the thaw bulb in the first 10 years, and then increases slowly when the thaw bulb extends to the weathered granite. Differential thaw settlement will cause a significant change in the deformation and stress of the pipeline near the interface of strong and weak thaw settlement zones. When the length ratio of strong and weak thaw settlement zones is 1, the maximum stress of the pipeline with a thickness of 16 mm is approximately 45% of the allowable stress of X65 steel, and the pipeline remains safe for 30 years. However, the potential failure of the pipeline should be considered due to the continued ground thawing and warming and pipe material aging. Forthcoming research on this topic is needed to evaluate more carefully the structural integrity of buried pipelines in cold regions.

The Influence of Selected Plant Growth Regulators and Carbohydrates on In Vitro Shoot Multiplication and Bulbing of the Tulip (Tulipa L.).

The aim of this study was to check the effects of sugar type on the in vitro shoot multiplication of the tulip cultivar 'Heart of Warsaw' and the effects of paclobutrazol (PBZ) and 1-naphthylacetic acid (NAA) on the bulbing of previously multiplied shoots. In addition, the subsequent effects of previously used sugars on the in vitro bulb formation of this cultivar were checked. First, the optimum supplementation of Murashige and Skoog medium with plant growth regulators (PGRs) was selected for shoot multiplication. Of the six tested, the best results were obtained using a combination of 2iP 0.1 mg·L-1, NAA 0.1 mg·L-1, and mT 5.0 mg·L-1. The effects of different carbohydrates (sucrose, glucose, and fructose at 30 g·L-1 and a mixture of glucose and fructose at 15 g·L-1 each) on multiplication efficiency was then tested on this medium. The microbulb-forming experiment was carried out taking into consideration the effects of previously applied sugars, and at week 6, the agar medium was flooded with liquid medium containing NAA 2 mg·L-1, PBZ 1 mg·L-1, or medium without PGRs; in the first combination, the cultures were left on a single-phase medium, solidified with agar, as a control. After 2 months of treatment at 5 °C, the total number of microbulbs formed and the number and weights of mature microbulbs were assessed. The results obtained indicate the ability of using meta-topolin (mT) in tulip micropropagation and point to sucrose and glucose as the optimal carbohydrates for intensive shoot multiplication. The results lead to the conclusion that it is most advantageous to multiply tulip shoots on glucose medium and then to carry out cultures on a two-phase medium with PBZ, which results in a higher number of microbulbs and their faster maturation.

LhANS-rr1, LhDFR, and LhMYB114 Regulate Anthocyanin Biosynthesis in Flower Buds of Lilium 'Siberia'.

The bulb formation of Lilium is affected by many physiological and biochemical phenomena, including flower bud differentiation, starch and sucrose accumulation, photoperiod, carbon fixation, plant hormone transduction, etc. The transcriptome analysis of flower buds of Lilium hybrid 'Siberia' at different maturity stages showed that floral bud formation is associated with the accumulation of anthocyanins. The results of HPLC-MS showed that cyanidin is the major anthocyanin found in Lilium 'Siberia'. Transcriptome KEGG enrichment analysis and qRT-PCR validation showed that two genes related to flavonoid biosynthesis (LhANS-rr1 and LhDFR) were significantly up-regulated. The functional analysis of differential genes revealed that LhMYB114 was directly related to anthocyanin accumulation among 19 MYB transcription factors. Furthermore, the qRT-PCR results suggested that their expression patterns were very similar at different developmental stages of the lily bulbs. Virus-induced gene silencing (VIGS) revealed that down-regulation of LhANS-rr1, LhDFR, and LhMYB114 could directly lead to a decrease in anthocyanin accumulation, turning the purple phenotype into a white color. Moreover, this is the first report to reveal that LhMYB114 can regulate anthocyanin accumulation at the mature stage of lily bulbs. The accumulation of anthocyanins is an important sign of lily maturity. Therefore, these findings have laid a solid theoretical foundation for further discussion on lily bulb development in the future.

Phenotypic and genotypic landscape of PROKR2 in neuroendocrine disorders.

Prokineticin receptor 2 (PROKR2) encodes for a G-protein-coupled receptor that can bind PROK1 and PROK2. Mice lacking Prokr2 have been shown to present abnormal olfactory bulb formation as well as defects in GnRH neuron migration. Patients carrying mutations in PROKR2 typically present hypogonadotropic hypogonadism, anosmia/hyposmia or Kallmann Syndrome. More recently variants in PROKR2 have been linked to several other endocrine disorders. In particular, several patients with pituitary disorders have been reported, ranging from mild phenotypes, such as isolated growth hormone deficiency, to more severe ones, such as septo-optic dysplasia. Here we summarize the changing landscape of PROKR2-related disease, the variants reported to date, and discuss their origin, classification and functional assessment.