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Objective: To examine the clinical phenotype and genetic deficiencies present in Chinese aniridia families with PAX6 haplotype deficiency., Methods: A comprehensive questionnaire and ophthalmological assessments were administered to both affected patients and unaffected relatives. The clinical feature analysis included the evaluation of visual acuity, intraocular pressure, slit-lamp anterior segment examination, fundus photography, and spectral domain optical coherence tomography. To identify the mutation responsible for aniridia, targeted next-generation sequencing was used as a beneficial technique., Results: A total of 4 mutations were identified, consisting of two novel frameshift mutations (c.314delA, p.K105Sfs*33 and c.838_845dup AACACACC, p.S283Tfs*85), along with two recurring nonsense mutations (c.307C>T, p.R103X and c.619A>T, p.K207*). Complete iris absence, macular foveal hypoplasia, and nystagmus were consistent in these PAX6 haplotype-deficient Chinese aniridia families, while corneal lesions, cataracts, and glaucoma exhibited heterogeneity both among the families and within the same family., Conclusion: In our study, two novel PAX6 mutations associated with aniridia were identified in Chinese families, which expanded the phenotypic and genotypic spectrum of PAX6 mutations. We also analyzed the clinical characteristics of PAX6 haplotype deficiency in Chinese aniridia families., (© 2024. Huazhong University of Science and Technology.)

Objective: Autosomal recessive bestrophinopathy (ARB), a retinal degenerative disease, is characterized by central visual loss, yellowish multifocal diffuse subretinal deposits, and a dramatic decrease in the light peak on electrooculogram. The potential pathogenic mechanism involves mutations in the BEST1 gene, which encodes Ca 2+ -activated Cl - channels in the retinal pigment epithelium (RPE), resulting in degeneration of RPE and photoreceptor. In this study, the complete clinical characteristics of two Chinese ARB families were summarized., Methods: Pacific Biosciences (PacBio) single-molecule real-time (SMRT) sequencing was performed on the probands to screen for disease-causing gene mutations, and Sanger sequencing was applied to validate variants in the patients and their family members., Results: Two novel mutations, c.202T>C (chr11:61722628, p.Y68H) and c.867+97G>A, in the BEST1 gene were identified in the two Chinese ARB families. The novel missense mutation BEST1 c.202T>C (p.Y68H) resulted in the substitution of tyrosine with histidine in the N-terminal region of transmembrane domain 2 of bestrophin-1. Another novel variant, BEST1 c.867+97G>A (chr11:61725867), located in intron 7, might be considered a regulatory variant that changes allele-specific binding affinity based on motifs of important transcriptional regulators., Conclusion: Our findings represent the first use of third-generation sequencing (TGS) to identify novel BEST1 mutations in patients with ARB, indicating that TGS can be a more accurate and efficient tool for identifying mutations in specific genes. The novel variants identified further broaden the mutation spectrum of BEST1 in the Chinese population., (© 2024. Huazhong University of Science and Technology.)

Key Message: Rice glycosyltransferase gene UGT2 was identified to play a crucial role in salt tolerance. The transcription factor OsbZIP23 was demonstrated to regulate the UGT2 expression under stress conditions. UDP-glycosyltransferases (UGTs) play key roles in modulating plant responses to environmental challenges. In this study, we characterized a novel glycosyltransferase, UGT2, which plays an important role in salt stress responses in rice (Oryza sativa L). We found that seedlings overexpressing UGT2 exhibited better growth than wild type in shoot and root under hydroponic culture with salt stress treatments, while ugt2ko mutant lines suffered much more growth inhibition. When the soil-grown UGT2 transgenic plants were subjected to salt stress, we also found that ugt2ko mutant lines were severely withered and most of them died, while the overexpression lines grew well and had higher survival rate. Compared with wild-type plants, UGT2 overexpression greatly increased the expression levels of the reactive oxygen species scavenging genes and stress-responsive genes. Furthermore, the upstream regulatory mechanism of the UGT2 gene was identified and we found that a bZIP transcription factor, OsbZIP23, can bind to the UGT2 promoter and enhance the UGT2 transcription levels. This work reveals that OsbZIP23-UGT2 module may play a major role in regulating the salt stress tolerance in rice., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Aim: To present the clinical manifestations of 5 autosomal dominant cone-rod dystrophy (adCORD) patients from two Chinese families with cone-rod homeobox (CRX) mutation (p.R41W), and to explore the clinical heterogeneity of adCORD with CRX mutation (p.R41W)., Methods: Interrogation and ophthalmological examinations were undertaken in all patients and unaffected members. Analysis of clinical features was performed by visual acuity, slit lamp examination, visual field examination, fundoscopy, autofluorescence and spectral domain optical coherence tomography. Targeted next-generation sequencing was applied as a useful tool to identify the causative mutation of CORD genes., Results: A CRX missense mutation c.121C>T was identified in all patients, resulting in an amino acid change from arginine acid to tryptophan (p.R41W). The patients presented with early onset, progressive and different severities with CORD., Conclusion: This is the first report of the clinical phenotype of CRX mutation (p.R41W) in Chinese families, and the mutation can lead to a wide range of various retinal phenotypes., (International Journal of Ophthalmology Press.)

Drought is one of the most important environmental constraints affecting plant growth and development and ultimately leads to yield loss. Uridine diphosphate (UDP)-dependent glycosyltransferases (UGTs) are believed to play key roles in coping with environmental stresses. In rice, it is estimated that there are more than 200 UGT genes. However, most of them have not been identified as their physiological significance. In this study, we reported the characterization of a putative glycosyltransferase gene UGT85E1 in rice. UGT85E1 gene is significantly upregulated by drought stress and abscisic acid (ABA) treatment. The overexpression of UGT85E1 led to an enhanced tolerance in transgenic rice plants to drought stress, while the ugt85e1 mutants of rice showed a more sensitive phenotype to drought stress. Further studies indicated that UGT85E1 overexpression induced ABA accumulation, stomatal closure, enhanced reactive oxygen species (ROS) scavenging capacity, increased proline and sugar contents, and upregulated expression of stress-related genes under drought stress conditions. Moreover, when UGT85E1 was ectopically overexpressed in Arabidopsis, the transgenic plants showed increased tolerance to drought as well as in rice. Our findings suggest that UGT85E1 plays an important role in mediating plant response to drought and oxidative stresses. This work may provide a promising candidate gene for cultivating drought-tolerant crops both in dicots and monocots., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Liu, Dong, Ma, Zhao, Liu, Li, Li and Hou.)

Maize is one of the major crops in the world; however, diseases caused by various pathogens seriously affect its yield and quality. The maize Rp1-D21 mutant (mt) caused by the intragenic recombination between two nucleotide-binding, leucine-rich repeat (NLR) proteins, exhibits autoactive hypersensitive response (HR). In this study, we integrated transcriptomic and metabolomic analyses to identify differentially expressed genes (DEGs) and differentially accumulated metabolites (DAMs) in Rp1-D21 mt compared to the wild type (WT). Genes involved in pathogen-associated molecular pattern (PAMP)-triggered immunity (PTI) and effector-triggered immunity (ETI) were enriched among the DEGs. The salicylic acid (SA) pathway and the phenylpropanoid biosynthesis pathway were induced at both the transcriptional and metabolic levels. The DAMs identified included lipids, flavones, and phenolic acids, including 2,5-DHBA O -hexoside, the production of which is catalyzed by uridinediphosphate (UDP)-dependent glycosyltransferase (UGT). Four maize UGTs ( ZmUGTs) homologous genes were among the DEGs. Functional analysis by transient co-expression in Nicotiana benthamiana showed that ZmUGT9250 and ZmUGT5174, but not ZmUGT9256 and ZmUGT8707, partially suppressed the HR triggered by Rp1-D21 or its N-terminal coiled-coil signaling domain (CC D21 ). None of the four ZmUGTs interacted physically with CC D21 in yeast two-hybrid or co-immunoprecipitation assays. We discuss the possibility that ZmUGTs might be involved in defense response by regulating SA homeostasis., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Ge, Wang, Lu, Zhao, Hou, Balint-Kurti and Wang.)

Recent studies have shown that global metabolic reprogramming is a common event in plant innate immunity; however, the relevant molecular mechanisms remain largely unknown. Here, we identified a pathogen-induced glycosyltransferase, UGT73C7, that plays a critical role in Arabidopsis disease resistance through mediating redirection of the phenylpropanoid pathway. Loss of UGT73C7 function resulted in significantly decreased resistance to Pseudomonas syringae pv. tomato DC3000, whereas constitutive overexpression of UGT73C7 led to an enhanced defense response. UGT73C7-activated immunity was demonstrated to be dependent on the upregulated expression of SNC1, a Toll/interleukin 1 receptor-type NLR gene. Furthermore, in vitro and in vivo assays indicated that UGT73C7 could glycosylate p-coumaric acid and ferulic acid, the upstream metabolites in the phenylpropanoid pathway. Mutations that lead to the loss of UGT73C7 enzyme activities resulted in the failure to induce SNC1 expression. Moreover, glycosylation activity of UGT73C7 resulted in the redirection of phenylpropanoid metabolic flux to biosynthesis of hydroxycinnamic acids and coumarins. The disruption of the phenylpropanoid pathway suppressed UGT73C7-promoted SNC1 expression and the immune response. This study not only identified UGT73C7 as an important regulator that adjusts phenylpropanoid metabolism upon pathogen challenge, but also provided a link between phenylpropanoid metabolism and an NLR gene., (© 2021 Society for Experimental Biology and John Wiley & Sons Ltd.)

Plants can reduce or eliminate the damage caused by herbicides and gain herbicide resistance, which is an important theoretical basis for the development of herbicide-resistant crops at this stage. Thus, discovering novel herbicide-resistant genes to produce diverse herbicide-resistant crop species is of great value. The glycosyltransferases that commonly exist in plant kingdom modify the receptor molecules to change their physical characteristics and biological activities, and thus possess an important potential to be used in the herbicide-resistance breeding. Here, we identified a novel herbicide-induced UDP-glycosyltransferase 91C1 (UGT91C1) from Arabidopsis thaliana and demonstrated its glucosylating activity toward sulcotrione, a kind of triketone herbicides widely used in the world. Overexpression of UGT91C1 gene enhanced the Arabidopsis tolerance to sulcotrione. While, ugt91c1 mutant displayed serious damage and reduced chlorophyll contents in the presence of sulcotrione, suggesting an important role of UGT91C1 in herbicide detoxification through glycosylation. Moreover, it was also noted that UGT91C1 can affect tyrosine metabolism by reducing the sulcotrione toxicity. Together, our identification of glycosyltransferase UGT91C1, as a potential gene conferring herbicide detoxification through glucosylation, may open up a new possibility for herbicide resistant breeding of crop plants and environmental phytoremediation., (Copyright © 2020 Elsevier Masson SAS. All rights reserved.)

The signaling network formed by external environmental signals and endogenous hormone signals is an important basis for the adaptive growth of plants. We recently identified a UDP-glucosyltransferase gene, UGT76F1 , which controls the glucosylation of auxin precursor IPyA and mediates light-temperature signaling to regulate auxin-dependent hypocotyl elongation in Arabidopsis. However, it is unclear whether UGT76F1 is involved in the adaptive growth of other tissues and whether it is related to the signaling of other hormones besides auxin. Here we investigated the petiole elongation of UGT76F1 overexpression lines and knockout mutant lines, and also studied the effects of UGT76F1 on BR signaling. Experimental results indicated that UGT76F1 is involved in the PIF4-mediated petiole growth under high temperature and that UGT76F1 is also related to the BR signaling in controlling hypocotyl growth. These results suggest that UGT76F1 may have a wider significance in the plant adaptations to surrounding environments.

Auxin is a class of plant hormone that plays a crucial role in the life cycle of plants, particularly in the growth response of plants to ever-changing environments. Since the auxin responses are concentration-dependent and higher auxin concentrations might often be inhibitory, the optimal endogenous auxin level must be closely controlled. However, the underlying mechanism governing auxin homeostasis remains largely unknown. In this study, a UDP-glycosyltransferase (UGT76F1) was identified from Arabidopsis thaliana , which participates in the regulation of auxin homeostasis by glucosylation of indole-3-pyruvic acid (IPyA), a major precursor of the auxin indole-3-acetic acid (IAA) biosynthesis, in the formation of IPyA glucose conjugates (IPyA-Glc). In addition, UGT76F1 was found to mediate hypocotyl growth by modulating active auxin levels in a light- and temperature-dependent manner. Moreover, the transcription of UGT76F1 was demonstrated to be directly and negatively regulated by PIF4, which is a key integrator of both light and temperature signaling pathways. This study sheds a light on the trade-off between IAA biosynthesis and IPyA-Glc formation in controlling auxin levels and reveals a regulatory mechanism for plant growth adaptation to environmental changes through glucosylation of IPyA., Competing Interests: The authors declare no competing interest.

Key Message: This study revealed that the Arabidopsis UGT75B1 plays an important role in modulating ABA activity by glycosylation when confronting stress environments. The cellular ABA content and activity can be tightly controlled in several ways, one of which is glycosylation by family 1 UDP-glycosyltransferases (UGTs). Previous analysis has shown UGT75B1 activity towards ABA in vitro. However, the biological role of UGT75B1 remains to be elucidated. Here, we characterized the function of UGT75B1 in abiotic stress responses via ABA glycosylation. GUS assay and qRT-PCR indicated that UGT75B1 is significantly upregulated by adverse conditions, such as osmotic stress, salinity and ABA. Overexpression of UGT75B1 in Arabidopsis leads to higher seed germination rates and seedling greening rates upon exposure to salt and osmotic stresses. In contrast, the big UGT75B1 overexpression plants are more sensitive under salt and osmotic stresses. Additionally, the UGT75B1 overexpression plants showed larger stomatal aperture and more water loss under drought condition, which can be explained by lower ABA levels examined in UGT75B1 OE plants in response to water deficit conditions. Consistently, UGT75B1 ectopic expression leads to downregulation of many ABA-responsive genes under stress conditions, including ABI3, ABI5 newly germinated seedlings and RD29A, KIN1, AIL1 in big plants. In summary, our results revealed that the Arabidopsis UGT75B1 plays an important role in coping with abiotic stresses via glycosylation of ABA.

Background: In cereal crop rice, auxin is known as an important class of plant hormone that regulates a plethora of plant growth and development. Glycosylation of auxin is known to be one of the important mechanisms mediating auxin homeostasis. However, the relevant auxin glucosyltransferase (GT) in rice still remains largely unknown., Results: In this study, using known auxin glucosyltransferases from other species as queries, twelve putative auxin UDP-glycosyltransferase (UGT) genes were cloned from rice and the one showing highest sequence similarity, named as OsIAGT1, was expressed as recombinant protein. In vitro enzymatic analysis showed that recombinant OsIAGT1 was capable of catalyzing glucosylation of IAA, IBA and other auxin analogs, and that OsIAGT1 is quite tolerant to a broad range of reaction conditions with peak activity at 30 °С and pH 8.0. OsIAGT1 showed favorite activity towards native auxins over artificially synthesized ones. Further study indicated that expression of OsIAGT1 can be upregulated by auxin in rice, and with OsIAGT1 overexpressing lines we confirmed that OsIAGT1 is indeed able to glucosylate IAA in vivo. Consistently, ectopic expression of OsIAGT1 leads to declined endogenous IAA content, as well as upregulated auxin synthesis genes and reduced expression of auxin-responsive genes, which likely leads to the reduced plant stature and root length in OsIAGT1 overexpression lines., Conclusion: Our result indicated that OsIAGT1 plays an important role in mediating auxin homeostasis by catalyzing auxin glucosylation, and by which OsIAGT1 regulates growth and development in rice.

The family Alvinocarididae is the monophyletic taxon which lives restrictively at chemosynthesis-based environments in the deep-sea. Here, for the first time, we report the complete mitogenome of the alvinocaridid vent shrimp Rimicaris variabilis from the North Fiji Basin. The mitogenome was 15,909 bp in length, with 65.6% AT content. Its protein-coding gene organization was typical of other alvinocaridid shrimps. Based on the phylogenetic tree, R. variabilis was most closely related to Shinkaicaris leurokolos , rather than with other Rimicaris species. To resolve this incongruence between traditional morphological classification and molecular analyses, further mitogenomic analysis of undetermined alvinocaridid taxa is necessary., Competing Interests: The authors report no conflicts of interest. The authors alone are responsible for the content and writing of the paper., (© 2019 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.)

Plant systemic acquired resistance (SAR) provides an efficient broad-spectrum immune response to pathogens. SAR involves mobile signal molecules that are generated by infected tissues and transported to systemic tissues. Methyl salicylate (MeSA), a molecule that can be converted to salicylic acid (SA), is an essential signal for establishing SAR, particularly under a short period of exposure to light after pathogen infection. Thus, the control of MeSA homeostasis is important for an optimal SAR response. Here, we characterized a uridine diphosphate-glycosyltransferase, UGT71C3, in Arabidopsis ( Arabidopsis thaliana ), which was induced mainly in leaf tissue by pathogens including Pst DC3000/ avrRpt2 ( Pseudomonas syringae pv tomato strain DC3000 expressing avrRpt2). Biochemical analysis indicated that UGT71C3 exhibited strong enzymatic activity toward MeSA to form MeSA glucosides in vitro and in vivo. After primary pathogen infection by Pst DC3000/ avrRpt2 , ugt71c3 knockout mutants exhibited more powerful systemic resistance to secondary pathogen infection than that of wild-type plants, whereas systemic resistance in UGT71C3 overexpression lines was compromised. In agreement, after primary infection of local leaves, ugt71c3 knockout mutants accumulated significantly more systemic MeSA and SA than that in wild-type plants. whereas UGT71C3 overexpression lines accumulated less. Our results suggest that MeSA glucosylation by UGT71C3 facilitates negative regulation of the SAR response by modulating homeostasis of MeSA and SA. This study unveils further SAR regulation mechanisms and highlights the role of glucosylation of MeSA and potentially other systemic signals in negatively modulating plant systemic defense., (© 2019 American Society of Plant Biologists. All Rights Reserved.)

To understand the origin, migration, and distribution of organisms across disjunct deep-sea vent habitats, previous studies have documented the population genetic structures of widely distributed fauna, such as gastropods, bivalves, barnacles, and squat lobsters. However, a limited number of investigations has been conducted in the Southwest Pacific Ocean, and many questions remain. In this study, we determined the population structure of the bythograeid crab Austinograea alayseae from three adjacent vent systems (Manus Basin, North Fiji Basin, and Tonga Arc) in the Southwest Pacific Ocean using the sequences of two mitochondrial genes (COI and 16S rDNA) and one nuclear gene (28S rDNA). Populations were divided into a Manus clade and a North Fiji-Tonga clade, with sequence divergence values in the middle of the barcoding gap for bythograeids. We inferred that hydrographic and/or physical barriers act on the gene flow of A. alayseae between the Manus and North Fiji basins. Austinograea alayseae individuals interact freely between the North Fiji Basin and the Lau Basin (Tonga Arc). Although further studies of genetic differentiation over a geological time scale, life-history attributes, and genome-based population genetics are needed to improve our understanding of the evolutionary history of A. alayseae, our results contribute to elucidating the phylogeny, evolution, and biogeography of bythograeids., Competing Interests: The authors have declared that no competing interests exist.

Background and Aims: Nowadays, the plant family 1 glycosyltransferases (UGTs) are attracting more and more attention since members of this family can improve the properties of secondary metabolites and have significantly enriched the chemical species in plants. Over the past decade, most studies on UGTs have been conducted in Arabidopsis thaliana and they were proved to play diverse roles during the plant life cycle. The Zea mays (maize) GT1 family comprises a large number of UDP-glycosyltransferase (UGT) members. However, their enzyme activities and the biological functions are rarely revealed. In this study, a maize flavonol glycosyltransferase, UFGT2, is identified and its biological role is characterized in detail., Methods: The UFGT2 enzyme activity, the flavonol and glycoside levels in planta were examined by high- performance liquid chromatography (HPLC) and liquid chromatography-mass spectrometry (LC-MS). The functions of UFGT2 in modifying flavonols, mediating flavonol accumulation and improving stress tolerance were analysed using two ufgt2 mutants and transgenic arabidopsis plants., Key Results: By in vitro enzyme assay, the maize UFGT2 was found to show strong activity towards two flavonols: kaemferol and quercetin. Two ufgt2 knockout mutants, Mu689 and Mu943, exhibited obvious sensitivity to salt and drought stresses. The endogenous quercetin and kaempferol glycosides, as well as the total flavonol levels were found to be substantially decreased in the two ufgt2 mutants, with declined H2O2-scavenging capacity. In contrast, ectopic expression of UFGT2 in arabidopsis led to increased flavonol contents and enhanced oxidative tolerance. Moreover, expression of typical stress-related genes in arabidopsis and maize were affected in UFGT2 overexpression plants or knockout mutants in response to abiotic stresses. UFGT2 was also transferred into the arabidopsis ugt78d2 mutant and it was found to recover the deficient flavonol glycoside pattern in the ugt78d2 mutant, which confirmed its catalysing activity in planta., Conclusion: It is demonstrated in our study that a maize glycosyltransferase, UFGT2, involved in modifying flavonols, contributes to improving plant tolerance to abiotic stresses.

The family Catophragmidae is one of the lower balanomorphs from traditional and recent multiple mitochondrial and nuclear markers molecular analysis. Here, we characterized the first mitogenome of the catophragmid barnacle Catomerus polymerus , which was 15,446 bp in length with a 68.3% AT content. The mitogenome had the typical pancrustacean gene arrangement, which was identical to the mitogenome configurations of the chthamalid Octomeris sp. and pachylasmatoid Eochionelasmus ohtai . On the mitogenomic tree, the catophragmid Catomerus polymerus formed an independent branch that was basal to the members of the superfamilies Tetraclitoidea and Balanoidea, which was inconsistent with previous findings., Competing Interests: The authors report no conflicts of interest. The authors alone are responsible for the content and writing of the paper., (© 2018 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.)

Salicylic acid (SA) plays a crucial role in plant innate immunity. The deployment of SA-associated immune responses is primarily affected by SA concentration, which is determined by a balance between SA biosynthesis and catabolism. However, the mechanisms regulating SA homeostasis are poorly understood. In this study, we characterized a unique UDP-glycosyltransferase, UGT76D1, which plays an important role in SA homeostasis and associated immune responses in Arabidopsis ( Arabidopsis thaliana ). Expression of UGT76D1 was induced by treatment with both the pathogen Pseudomonas syringae pv. tomato ( Pst ) DC3000 and SA. Overexpression of UGT76D1 resulted in high SA accumulation, significant up-regulation of pathogen-related genes, and a hypersensitive response (HR)-like lesion mimic phenotype. This HR-like phenotype was not observed following UGT76D1 overexpression in SA-deficient NahG transgenic or sid2 plants, suggesting that the phenotype is SA dependent. Biochemical assays showed that UGT76D1 glycosylated 2,3-dihydroxybenzoic acid (2,3-DHBA) and 2,5-dihydroxybenzoic acid (2,5-DHBA), the major catabolic forms of SA, to their Glc and Xyl conjugates in vitro and in vivo. Moreover, in a mutant background blocked in the formation of 2,3-DHBA and 2,5-DHBA, UGT76D1 overexpression did not cause a HR-like lesion mimic phenotype. Following infection with Pst DC3000, UGT76D1 knockout mutants displayed a delayed immune response, with reduced levels of DHBA glycosides and SA, and down-regulated SA synthase expression. By contrast, UGT76D1 overexpression lines showed an enhanced immune response and increased SA biosynthesis before and after pathogen infection. Thus, we propose that UGT76D1 plays an important role in SA homeostasis and plant immune responses by facilitating glycosylation of dihydroxybenzoic acids., (© 2018 American Society of Plant Biologists. All Rights Reserved.)

Although plant glycosyltransferases are thought to play important roles in growth and interaction with the environment, little is known about their physiological roles for most members of the plant glycosyltransferase family. We cloned and characterised an Arabidopsis glycosyltransferase gene, UGT76E11. Its in vivo physiological effects on flavonoid accumulation and plant tolerance to abiotic stresses were investigated. The UGT76E11 gene was up-regulated in transcription expression under stress conditions of salinity, drought and H 2 O 2 treatment. Transgenic plants ectopically overexpressing UGT76E11 showed substantially enhanced tolerance to salinity and drought at germination and during post-germination growth. Enzyme activity of UGT76E11 to glucosylate quercetin and other flavonoids was confirmed. Ectopic expression of UGT76E11 resulted in significantly increased flavonoid content in transgenic plants compared to wild type, suggesting a contribution of UGT76E11 to modulation of flavonoid metabolism. Consistent with this result, several biosynthesis genes in the flavonoid pathway were clearly up-regulated in transgenic plants. Furthermore, overexpression of UGT76E11 also enhanced the scavenging capacity for ROS and increased expression levels of a number of stress-related genes. Based on these results, we suggest that the glycosyltransferase UGT76E11 plays an important role in modulating flavonoid metabolism and enhancing plant adaptation to environmental stresses. Our findings might allow use of glycosyltransferase UGT76E11 in crop improvement, towards both enhanced stress tolerance and increased flavonoid accumulation., (© 2017 German Society for Plant Sciences and The Royal Botanical Society of the Netherlands.)

The hitherto suborder Verrucomorpha contains asymmetrical barnacles of two groups: the true Verrucomorpha ( Eoverruca  + Verrucidae) and the Neoverrucidae. Here, we determined the mitochondrial genome (mitogenome) of Altiverruca navicula , a true Verrucomorpha species. The mitogenome was 15,976 base pairs in length and had the typical pancrustacean gene arrangement. Its protein-coding genes were very similar to those of other thoracican species in terms of length, AT content, and start and stop codons. In phylogenetic trees constructed with 13 protein-coding genes, A . navicula was positioned at an ancestral node of sessile barnacles, consistent with the findings of previous studies., Competing Interests: The authors report no conflicts of interest. The authors alone are responsible for the content and writing of the paper., (© 2017 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.)

Key Message: Ectopic expression of auxin glycosyltransferase UGT84A2 in Arabidopsis can delay flowering through increased indole-3-butyric acid and suppressed transcription of ARF6, ARF8 and flowering-related genes FT, SOC1, AP1 and LFY. Auxins are critical regulators for plant growth and developmental processes. Auxin homeostasis is thus an important issue for plant biology. Here, we identified an indole-3-butyric acid (IBA)-specific glycosyltransferase, UGT84A2, and characterized its role in Arabidopsis flowering development. UGT84A2 could catalyze the glycosylation of IBA, but not indole-3-acetic acid (IAA). UGT84A2 transcription expression was clearly induced by IBA. When ectopically expressing in Arabidopsis, UGT84A2 caused obvious delay in flowering. Correspondingly, the increase of IBA level, the down-regulation of AUXIN RESPONSE FACTOR 6 (ARF6) and ARF8, and the down-regulation of flowering-related genes such as FLOWERING LOCUS T (FT), SUPPRESSOR OF OVEREXPRESSION OF CO1(SOC1), APETALA1 (AP1), and LEAFY(LFY) were observed in transgenic plants. When exogenously applying IBA to wild-type plants, the late flowering phenotype, the down-regulation of ARF6, ARF8 and flowering-related genes recurred. We examined the arf6arf8 double mutants and found that the expression of flowering-related genes was also substantially decreased in these mutants. Together, our results suggest that glycosyltransferase UGT84A2 may be involved in flowering regulation through indole-3-butyric acid-mediated transcriptional repression of ARF6, ARF8 and downstream flowering pathway genes.

The disialic acid-containing glycosphingolipid GD3 recruited membrane transglutaminase 2 (TG2) as a signaling molecule for erythroid differentiation in human chronic myelogenous leukemia (CML) K562 cells. The α1-adrenergic receptor (α1-AR)/TG2-mediated signaling pathway regulated GD3 functions, including gene expression and production, to differentiate CML K562 cells into erythroid lineage cells. Epinephrine, an AR agonist, increased membrane recruitment as well as GTP-photoaffinity of TG2, inducing GD3 synthase gene expression. Epinephrine activated PI3K/Akt signaling and GTPase downstream of TG2 activated Akt. The coupling of TG2 and GD3 production was specifically suppressed by prazosin (α1-AR antagonist), but not by propranolol (β-AR antagonist) or rauwolscine (α2-AR antagonist), indicating α1-AR specificity. Small interfering RNA (siRNA) experiment results indicated that the α1-AR/TG2-mediated signaling pathway activated PKCs α and δ to induce GD3 synthase gene expression. Transcription factors CREB, AP-1, and NF-κB regulated GD3 synthase gene expression during α1-AR-induced differentiation in CML K562 cells. In addition, GD3 synthase gene expression was upregulated in TG2-transfected cells via α1-AR with expression of erythroid lineage markers and benzidine-positive staining. α1-AR/TG2 signaling pathway-directed GD3 production is a crucial step in erythroid differentiation of K562 cells and GD3 interacts with α1-AR/TG2, inducing GD3/α1-AR/TG2-mediated erythroid differentiation. These results suggest that GD3, which acts as a membrane mediator of erythroid differentiation in CML cells, provides a therapeutic avenue for leukemia treatment., Competing Interests: CONFLICTS OF INTEREST The authors did not declare any competing interests in this study.

Glycosyltransferase (GT) family-1, the biggest GT family in plants, typically participates in modification of small molecules and affects many aspects during plant development. In Arabidopsis thaliana, although some UDP glycosyltransferases (UGTs) of family-1 have been functionally characterized, functions of most the UGTs remain unknown or fragmentary. Here, we report data for the Arabidopsis UGT87A2, a stress-regulated GT. We found that UGT87A2 could be dramatically induced by salinity, osmotic stress, drought and ABA. Overexpression of UGT87A2 (87A2OE) leads to accelerated germination and greening, higher survival rate as well as increased root length against abiotic stresses compared with those of wild-type (WT) plants. In addition, we observed lower water loss rate in the 87A2OE plants due to smaller stomatal apertures. The transgenic plants also showed reduced levels of H 2 O 2 and superoxide under low water status compared with those of WT plants. Consistently, function loss of UGT87A2 in ugt87a2 knockout lines resulted in opposite performances under these conditions. A transcriptome profiling revealed that 121 genes were differentially regulated upon UGT87A2 overexpression, and a large number of stress-induced genes were upregulated in UGT87A2 overexpression plants. Expression of seven genes among them were assessed by quantitative real-time polymerase chain reaction (qRT-PCR), including CPK32, CYP81F2, MYB96, DREB2A, FBS1, PUB23 and RAV2 under both control and stress treatments, and the results greatly validated our transcriptome data. Taken together, our findings support an explicit role of UGT87A2 in adaptation to abiotic stresses., (© 2016 Scandinavian Plant Physiology Society.)

The plant family 1 UDP-glycosyltransferases (UGTs) are the biggest GT family in plants, which are responsible for transferring sugar moieties onto a variety of small molecules, and control many metabolic processes; however, their physiological significance in planta is largely unknown. Here, we revealed that two Arabidopsis glycosyltransferase genes, UGT79B2 and UGT79B3, could be strongly induced by various abiotic stresses, including cold, salt and drought stresses. Overexpression of UGT79B2/B3 significantly enhanced plant tolerance to low temperatures as well as drought and salt stresses, whereas the ugt79b2/b3 double mutants generated by RNAi (RNA interference) and CRISPR-Cas9 strategies were more susceptible to adverse conditions. Interestingly, the expression of UGT79B2 and UGT79B3 is directly controlled by CBF1 (CRT/DRE-binding factor 1, also named DREB1B) in response to low temperatures. Furthermore, we identified the enzyme activities of UGT79B2/B3 in adding UDP-rhamnose to cyanidin and cyanidin 3-O-glucoside. Ectopic expression of UGT79B2/B3 significantly increased the anthocyanin accumulation, and enhanced the antioxidant activity in coping with abiotic stresses, whereas the ugt79b2/b3 double mutants showed reduced anthocyanin levels. When overexpressing UGT79B2/B3 in tt18 (transparent testa 18), a mutant that cannot synthesize anthocyanins, both genes fail to improve plant adaptation to stress. Taken together, we demonstrate that UGT79B2 and UGT79B3, identified as anthocyanin rhamnosyltransferases, are regulated by CBF1 and confer abiotic stress tolerance via modulating anthocyanin accumulation., (© 2016 The Authors The Plant Journal © 2016 John Wiley & Sons Ltd.)

Glycosylation of monolignols has been found to be widespread in land plants since the 1970s. However, whether monolignol glycosylation is crucial for cell wall lignification and how it exerts effects are still unknown. Here, we report the identification of a mutant ugt72b1 showing aggravated and ectopic lignification in floral stems along with arrested growth and anthocyanin accumulation. Histochemical assays and thioacidolysis analysis confirmed the enhanced lignification and increased lignin biosynthesis in the ugt72b1 mutant. The loss of UDP-glycosyltransferase UGT72B1 function was responsible for the lignification phenotype, as demonstrated by complementation experiments. Enzyme activity analysis indicated that UGT72B1 could catalyze the glucose conjugation of monolignols, especially coniferyl alcohol and coniferyl aldehyde, which was confirmed by analyzing monolignol glucosides of UGT72B1 transgenic plants. Furthermore, the UGT72B1 gene was strongly expressed in young stem tissues, especially xylem tissues. However, UGT72B1 paralogs, such as UGT72B2 and UGT72B3, had weak enzyme activity toward monolignols and weak expression in stem tissues. Transcriptomic profiling showed that UGT72B1 knockout resulted in extensively increased transcript levels of genes involved in monolignol biosynthesis, lignin polymerization and cell wall-related transcription factors, which was confirmed by quantitative real-time PCR assays. These results provided evidence that monolignol glucosylation catalyzed by UGT72B1 was essential for normal cell wall lignification, thus offering insight into the molecular mechanism of cell wall development and cell wall lignification., (© 2016 The Authors The Plant Journal © 2016 John Wiley & Sons Ltd.)

Retinitis pigmentosa (RP) describes a group of inherited retinopathies that are characterized by the progressive degeneration of photoreceptor neurons, which causes night blindness, a reduction in the peripheral visual field and decreased visual acuity. More than 50 RP-related genes have been identified. In the present study, we analysed a Chinese family with autosomal recessive RP. We identified a compound heterozygous mutation, c.265delC and c.1537G>A, in CNGA1 using targeted next-generation sequencing (NGS) of RP-causing genes. The mutations were validated in the family members by Sanger sequencing. The mutations co-segregated with the RP phenotype and were absent from ethnically-matched control chromosomes. The mutant (mut) CNGA1 p.(G513R) protein caused by the mis-sense novel mutation c.1537G>A was expressed in vitro. The mut CNGA1 p.(G513R) protein was largely retained inside the cell rather than being targeted to the plasma membrane, suggesting the absence of cGMP-gated cation channels in the plasma membrane would be deleterious to rod photoreceptors, leading lead to RP., (© 2016 Authors.)

The formation of auxin glucose conjugate is proposed to be one of the molecular modifications controlling auxin homeostasis. However, the involved mechanisms and relevant physiological significances are largely unknown or poorly understood. In this study, Arabidopsis UGT75D1 was at the first time identified to be an indole-3-butyric acid (IBA) preferring glycosyltransferase. Assessment of enzyme activity and IBA conjugates in transgenic plants ectopically expressing UGT75D1 indicated that the UGT75D1 catalytic specificity was maintained in planta. It was found that the expression pattern of UGT75D1 was specific in germinating seeds. Consistently, we found that transgenic seedlings with over-produced UGT75D1 exhibited smaller cotyledons and cotyledon epidermal cells than the wild type. In addition, UGT75D1 was found to be up-regulated under mannitol, salt and ABA treatments and the over-expression lines were tolerant to osmotic and salt stresses during germination, resulting in an increased germination rate. Quantitative RT-PCR analysis revealed that the mRNA levels of ABA INSENSITIVE 3 (ABI3) and ABI5 gene in ABA signaling were substantially down-regulated in the transgenic lines under stress treatments. Interestingly, AUXIN RESPONSE FACTOR 16 (ARF16) gene of transgenic lines was also dramatically down-regulated under the same stress conditions. Since ARF16 functions as an activator of ABI3 transcription, we supposed that UGT75D1 might play a role in stress tolerance during germination through modulating ARF16-ABI3 signaling. Taken together, our work indicated that, serving as the IBA preferring glycosyltransferase but distinct from other auxin glycosyltransferases identified so far, UGT75D1 might be a very important player mediating a crosstalk between cotyledon development and stress tolerance of germination at the early stage of plant growth.

Background: This study investigated whether functional neuronavigation can be used to remove lesions in the lateral ventricle while preserving patients' neurologic functionality., Methods: A total of 60 patients with lateral ventricular meningiomas were divided into study and control groups (n = 30 each). Diffusion tensor and blood oxygenation level-dependent functional magnetic resonance imaging were used for fiber tracking and eloquent cortex localization, respectively, in the study group. The surgical approach was based on coregistered data sets from 3-D lesion and brain structure reconstructions. Patients in the control group underwent anatomic neuronavigation-guided surgery. The patients' demographics, degree of resection, visual field, language score, movement, preoperative and postoperative Karnofsky Performance Status (KPS) scores, and surgical complications were recorded., Results: Tumors were completely removed in both groups. Patients in the study group had a higher rate of visual field preservation than controls (P = 0.01). The two groups had similar motor and language functions after surgery, except that fewer cases of transient aphasia were observed in the former (P < 0.05). KPS scores for the study and control groups were 80 (70-80) and 70 (60-70), respectively (P < 0.01), at 2 weeks and 90 (80-100) and 85 (70-90), respectively (P = 0.022), at 3 months after surgery., Conclusions: Functional neuronavigation preserved neurologic functionality and was especially beneficial for protecting optical functionality and for the rapid recovery of patients., (Copyright © 2015 Elsevier Inc. All rights reserved.)

The Arabidopsis uridine diphosphate (UDP)-glycosyltransferase 76C2 (UGT76C2), a member of family 1 UGTs, is described as a cytokinin glycosyltransferase. In this study, we demonstrate a novel role of UGT76C2 in response to water deficit. QRT-PCR assay identified that the expression of this gene was downregulated by drought, osmotic stress and abscisic acid (ABA). Compared with wild type (WT) plants, transgenic lines ectopically expressing UGT76C2 exhibited reduced tolerance to ABA and osmotic stress during postgermination growth, while enhanced adaptation to drought stress at mature stage. Consistently, the ugt76c2 mutant plants showed opposite responses to these conditions. To explore the possible mechanisms of UGT76C2 contributing to drought stress adaptation, six stress inducible genes including DREB2A, RD22, RD29B, LEA, COR47 and KIN1 were detected, which showed significant upregulation in UGT76C2 overexpression plants under drought stress. Besides, five cytokinin marker genes AHK2, AHK3, AHK4, ARR1 and ARR2 were also evaluated, which showed less induced in UGT76C2 overexpression plants in response to drought stress. Our results reveal that UGT76C2, as a cytokinin glycosyltransferase, is involved in the plant response to drought stress and might represent novel cues in abiotic stress adaptation., (Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.)

Purpose: The aim of this study is to examine whether or not myocilin (MYOC) genetic variations are associated with susceptibility to primary angle-closure glaucoma (PACG) in the Han Chinese population., Methods: Four single-nucleotide polymorphisms (SNPs)-rs235913, rs183532, rs12076134, and rs235875-in the MYOC gene were genotyped in 212 adult patients with PACG and 255 age-, sex-, and ethnic-matched healthy controls by using a polymerase chain reaction-restriction fragment length polymorphism assay. Data were analyzed by chi-square analysis., Results: The four SNPs in the MYOC gene were in the Hardy-Weinberg equilibrium in all the subjects. The frequencies of A allele rs183532 were significantly different between the PACG patients and the controls (0.238 vs. 0.169, p=0.008; OR=1.541; 95% CI: 1.117-2.127). The frequencies of the AA genotype and A allele of rs235913 were increased in PACG patients compared with controls, but the difference was not significant (p=0.037, p=0.017, respectively). A comparison of the distributions of the genotypes and alleles of rs12076134 and rs235875 showed no statistically significant differences between the PACG patients and the controls (p>0.05). Haplotype analysis indicated that the frequency of the AATG and AATA haplotypes was significantly higher for PACG patients than for control subjects (both p<0.001). However, the frequency of CGGA and CGTG haplotypes was lower for PACG patients than for control subjects (p<0.001)., Conclusions: Our study suggests that rs183532 is associated with an increased risk of PACG in the Chinese Han population.

Background: Diabetic retinopathy, the main microvascular complications of diabetes and one of the leading causes of blindness worldwide. Interesting reports on the role of inflammatory/proangiogenic high mobility group 1 (HMGB-1) cytokine and phospholipases A2 (PLA2) in neovascularization have diverted our concentration to reveal whether HMGB-1 and PLA2 plays role in diabetic retinopathy., Methods: We performed our study in streptozotocin (STZ)-induced diabetic rat model. The expression levels of the cytokines, chemokines, and cell adhesion molecules in retinal tissues were evaluated by quantitative RT-PCR. HMGB-1 and PLA2 protein levels along with VEGF, TNF-α, IL-1β and ICAM-1 levels were also measured., Results: We observed the retinal pericytes, endothelial injury/death and breakdown of blood-retinal barrier (BRB). The protein expression of HMGB-1, PLA2 and IL-1β were significantly increased in micro vessels from retina of diabetic rats. Diabetic rats had also high retinal levels of VEGF, ICAM-1 and TNF-α. Further investigation revealed that pericyte death is mediated by HMGB-1-induced cytotoxic activity of glial cells, while HMGB-1 can directly mediate endothelial cell death. Similarly, increased expression of PLA2 represents the diabetic mediated alteration of BRB, perhaps up regulating the VEGF., Conclusions: Our data suggest that HMGB-1 and PLA2 involved in retinal pericyte and endothelial injury and cell death in diabetic retinopathy. From this study, we suggest that HMGB-1 and PLA2 may be interesting targets in managing diabetic retinopathy.

Most current metabolic engineering applications rely on the inactivation of unwanted reactions and the amplification of product-oriented reactions. All of the biochemical reactions involved with cellular metabolism are tightly coordinated with the electron flow, which depends on the cellular energy status. Thus, the cellular metabolic flux can be controlled either by modulation of the electron flow or the regulation of redox reactions. This study analyzed the genome-wide anaerobic fermentation products of 472 Escherichia coli single gene knockouts, which comprised mainly of dehydrogenases, oxidoreductases, and redox-related proteins. Many metabolic pathways that were located far from anaerobic mixed-acid fermentation significantly affected the profiles of lactic acid, succinic acid, acetic acid, formic acid, and ethanol. Unexpectedly, D-lactate overproduction was determined by a single gene deletion in dehydrogenases (e.g., guaB, pyrD, and serA) involved with nucleotide and amino acid metabolism. Furthermore, the combined knockouts of guaB, pyrD, serA, fnr, arcA, or arcB genes, which are involved with anaerobic transcription regulation, enhanced D-lactate overproduction. These results suggest that the anaerobic fermentation profiles of E. coli can be tuned via the disruption of peripheral dehydrogenases in anaerobic conditions., (Copyright © 2013 Elsevier Inc. All rights reserved.)

The recent nucleic acid sequencing revolution driven by shotgun and high-throughput technologies has led to a rapid increase in the number of sequences for microbial communities. The availability of 16S ribosomal RNA (rRNA) gene sequences from a multitude of natural environments now offers a unique opportunity to study microbial diversity and community structure. The large volume of sequencing data however makes it time consuming to assign individual sequences to phylotypes by searching them against public databases. Since ribosomal sequences have diverged across prokaryotic species, they can be grouped into clusters that represent operational taxonomic units. However, available clustering programs suffer from overlap of sequence spaces in adjacent clusters. In natural environments, gene sequences are homogenous within species but divergent between species. This evolutionary constraint results in an uneven distribution of genetic distances of genes in sequence space. To cluster 16S rRNA sequences more accurately, it is therefore essential to select core sequences that are located at the centers of the distributions represented by the genetic distance of sequences in taxonomic units. Based on this idea, we here describe a novel sequence clustering algorithm named CLUSTOM that minimizes the overlaps between adjacent clusters. The performance of this algorithm was evaluated in a comparative exercise with existing programs, using the reference sequences of the SILVA database as well as published pyrosequencing datasets. The test revealed that our algorithm achieves higher accuracy than ESPRIT-Tree and mothur, few of the best clustering algorithms. Results indicate that the concept of an uneven distribution of sequence distances can effectively and successfully cluster 16S rRNA gene sequences. The algorithm of CLUSTOM has been implemented both as a web and as a standalone command line application, which are available at http://clustom.kribb.re.kr.

Aim: To investigate the role of heparanase-1 in laser-induced choroidal neovascularization (CNV)., Methods: Experimental CNV was induced by krypton laser photocoagulation in 15 male Brown Norway rats. Fundus fluorescein angiography and histopathological examination were performed in observing the CNV development. The expression and distribution of heparanase-1 protein in the laser lesions were determined by immunohistochemistry and western blotting analysis., Results: The success rate of laser induced CNV was approximately 75% on 3-4 weeks after laser photocoagulation. The protein levels of heparanase-1 increased significantly in the retina-choroidal complex of CNV models when compared to normal rat eyes (P<0.01). Immunostaining confirmed strong heparanase-1 expressions in all laser lesions, and it displayed to be highest at the newly formed blood vessels within the fibrovascular complex in the subretinal space., Conclusion: Heparanase-1 is closely involved in the development of laser induced CNV.

Auxin is one type of phytohormones that plays important roles in nearly all aspects of plant growth and developmental processes. The glycosylation of auxins is considered to be an essential mechanism to control the level of active auxins. Thus, the identification of auxin glycosyltransferases is of great significance for further understanding the auxin regulation. In this study, we biochemically screened the group L of Arabidopsis thaliana glycosyltransferase superfamily for enzymatic activity toward auxins. UGT74D1 was identified to be a novel auxin glycosyltransferase. Through HPLC and LC-MS analysis of reaction products in vitro by testing eight substrates including auxins and other compounds, we found that UGT74D1 had a strong glucosylating activity toward indole-3-butyric acid [IBA], indole-3-propionic acid [IPA], indole-3-acetic acid [IAA] and naphthaleneacetic acid [NAA], catalyzing them to form corresponding glucose esters. Biochemical characterization showed that this enzyme had a maximum activity in HEPES buffer at pH 6.0 and 37°C. In addition, the enzymatic activity analysis of crude protein and the IBA metabolite analysis from transgenic Arabidopsis plants overexpressing UGT74D1 gene were also carried out. Experimental results indicated that over-production of the UGT74D1 in plants indeed led to increased level of the glucose conjugate of IBA. Moreover, UGT74D1 overexpression lines displayed curling leaf phenotype, suggesting a physiological role of UGT74D1 in affecting the activity of auxins. Our current data provide a new target gene for further genetic studies to understand the auxin regulation by glycosylation in plants.

Cytokinins are master regulators of plant growth and development. The glucosyltransferase UGT76C1 capable of N-glucosylation of different cytokinins at the N(7)- and N(9)-position was previously identified in Arabidopsis thaliana, but its physiological relevance in plants remains unclear. In the present work, we investigated the physiological characteristics of UGT76C1 mutant (ugt76c1) and its overexpressors. Under normal growth conditions, although ugt76c1 plants and UGT76C1 overexpressors did not display obvious phenotypic alteration, ugt76c1 plants significantly reduced the accumulation of cytokinin N-glucosides, whereas UGT76C1 overexpressors increased cytokinin N-glucosides. Unexpectedly, the concentrations of free forms of cytokinins (mainly trans-zeatin and N(6)-isopentenyladenine) were comparable to those of the wild type. Upon application of exogenous cytokinin, the mutant showed the same tendency of more sensitive cytokinin response in primary root elongation, chlorophyll retention and anthocyanin accumulation. In contrast, overexpressors showed a tendency of less sensitive cytokinin response in most tests. Furthermore, cytokinin-related genes were investigated for their expression; and the expression levels of AHK3, ARR1, CYP735A2 and LOG2 noticeably changed in ugt76c1 plants, suggesting that plants employ a set of cytokinin regulation mechanisms to coordinate the loss-of-function of UGT76C1. Tissue-specific expression of UGT76C1 showed a high level of expression in germinating seeds and young seedlings. Taken together, our data suggest that the glucosyltransferase UGT76C1 could finely modulate cytokinin responses in planta via N-glucosylation of cytokinins., (Copyright © 2013 Elsevier Masson SAS. All rights reserved.)

Trans-zeatin is a kind of cytokinins that plays a crucial role in plant growth and development. The master trans-zeatin O-glucosyltransferase of Arabidopsis thaliana, UGT85A1, has been previously identified through biochemical approach. To determine the in planta role of UGT85A1 gene, the characterization of transgenic Arabidopsis plants overexpressing UGT85A1 was carried out. Under normal conditions, transgenic Arabidopsis did not display clearly altered phenotypes. A remarkable alteration is that the accumulation level of the trans-zeatin O-glucosides was significantly increased in UGT85A1 overexpressing transgenic Arabidopsis, while other forms of cytokinins kept the similar concentrations compared to the wild type. When treated with exogenously applied trans-zeatin, UGT85A1 overexpressing Arabidopsis showed much less sensitivity to trans-zeatin in primary root elongation and lateral root formation. Meanwhile, the chlorophyll content of detached leaves of transgenic Arabidopsis was much lower than wild type. Studies of spatial-temporal expression patterns showed that UGT85A1 was mainly expressed in the early seedlings and developing seeds. Analysis of subcellular localization suggested that UGT85A1 was localized to cytoplasm and nucleus. Taken together, our data suggest that overexpression of Arabidopsis glucosyltransferase UGT85A1 influences trans-zeatin homeostasis and trans-zeatin responses likely through O-glucosylation in planta.

Abiotic stresses greatly influence plant growth and productivity. While glycosyltransferases are widely distributed in plant kingdom, their biological roles in response to abiotic stresses are largely unknown. In this study, a novel Arabidopsis glycosyltransferase gene UGT85A5 was identified as significantly induced by salt stress. Ectopic expression of UGT85A5 in tobacco enhanced the salt stress tolerance in the transgenic plants. There were higher seed germination rates, better plant growth and less chlorophyll loss in transgenic lines compared to wild type plants under salt stress. This enhanced tolerance of salt stress was correlated with increased accumulations of proline and soluble sugars, but with decreases in malondialdehyde accumulation and Na(+)/K(+) ratio in UGT85A5-expressing tobacco. Furthermore, during salt stress, expression of several carbohydrate metabolism-related genes including those for sucrose synthase, sucrose-phosphate synthase, hexose transporter and a group2 LEA protein were obviously upregulated in UGT85A5-expressing transgenic plants compared with wild type controls. Thus, these findings suggest a specific protective role of this glycosyltransferase against salt stress and provide a genetic engineering strategy to improve salt tolerance of crops.

• Family 1 glycosyltransferases comprise the greatest number of glycosyltransferases found in plants. The widespread occurrence and diversity of glycosides throughout the plant kingdom underscore the importance of these glycosyltransferases. • Here, we describe the identification and characterization of a late-flowering Arabidopsis (Arabidopsis thaliana) mutant, in which a putative family 1 glycosyltransferase gene, UGT87A2, was disrupted. The role and possible mechanism of UGT87A2 in the regulation of flowering were analyzed by molecular, genetic and cellular approaches. • The ugt87a2 mutant exhibited late flowering in both long and short days, and its flowering was promoted by vernalization and gibberellin. Furthermore, the mutant flowering phenotype was rescued by the wild-type UGT87A2 gene in complementation lines. Interestingly, the expression of the flowering repressor FLOWERING LOCUS C was increased substantially in the mutant, but decreased to the wild-type level in complementation lines, with corresponding changes in the expression levels of the floral integrators and floral meristem identity genes. The expression of UGT87A2 was developmentally regulated and its protein products were distributed in both cytoplasm and nucleus. • Our findings imply that UGT87A2 regulates flowering time via the flowering repressor FLOWERING LOCUS C. These data highlight an important role for the family 1 glycosyltransferases in the regulation of plant flower development., (© 2012 The Authors. New Phytologist © 2012 New Phytologist Trust.)

There are three classical problems at the chromosome level in cytogenetics, namely the formation mechanisms and effects of Barr body, polytenic chromosome, and lampbrush chromosome. Teachers and researchers keep sustaining attention to the Barr body because of the relationships between Barr body and the X chromosome dosage compensation effect in mammals, the human sex identification, and some human diseases. In our genetics teaching practice, we tried the case-based teaching method. We introduced the classical problems and research progress of the Barr body, as a line, into partial sections of our genetics teaching contents such as sex-linked genetic analysis, eukaryotic gene expression regulation, cancer genetic analysis, and genetic experiments. Finally, it will form a comprehensive summary of related knowledge of genetics through class discussion on the Barr body. We found that this teaching method can not only optimize the teaching contents of genetics, consolidate and widen students' basic knowledge, and help student to form the systemic and developmental views of a classical genetics problem, but also inspire students' interest in life sciences. Good teaching results have been achieved.

Family 1 glycosyltransferases catalyse the glycosylation of small molecules and play an important role in maintaining cell homeostasis and regulating plant growth and development. In this study, a putative glycosyltransferase gene of family 1, PtGT1, was cloned from poplar (Populus tomentosa Carr.). Sequence analysis showed that this gene encodes a protein of 481 amino acid residues with a conserved PSPG box at its C-terminal, suggesting that it is active in the glycosylation of plant secondary products. The PtGT1 gene was expressed in poplar stems and leaves, with a particularly high expression level in elongating stems. Transgenic tobacco plants ectopically over-expressing PtGT1 were obtained and phenotypes were analysed. Wiesner and Mäule staining showed that stem xylem of transgenic tobacco plants stained more strongly than controls. Measurement of the Klason lignins showed much higher lignin content in the transgenic lines than in control plants. Furthermore, the ectopic over-expression of PtGT1 in tobacco resulted in an early flowering phenotype. These findings offer a possible starting point towards better understanding of the function of poplar PtGT1, and provide a novel strategy for lignin engineering and flowering control in plants through the genetic manipulation of a poplar glycosyltransferase gene.

Adeno-associated virus vector plasmid carrying the expression cassette of brain-derived neurotrophic factor (BDNF), pAAV-BDNF, was constructed and packaged into recombinant adeno-associated virus (rAAV-BDNF). The rAAV-BDNF was intravitreally injected into streptzotocin (STZ)-induced diabetic Sprague-Dawley (SD) Rats. Data showed that over-expression of BDNF could increase alive retinal ganglion cell (RGC) number and improve its function in streptzotocin(STZ)-induced diabetic rats, which might be a new method to treat diabetic neuropathy and retinopathy.

Cytokinins are a class of phytohormones that play a crucial role in plant growth and development. The gene UGT76C2 encoding cytokinin N-glucosyltransferase of Arabidopsis thaliana has been previously identified. To determine the in planta role of UGT76C2 in cytokinin metabolism and response, we analyzed the phenotypes of its loss-of-function mutant (ugt76c2) and its overexpressors. The accumulation level of the cytokinin N-glucosides was significantly decreased in ugt76c2, but substantially increased in UGT76C2 overexpressors compared with the wild type. When treated with exogenously applied cytokinin, ugt76c2 showed more sensitivity and UGT76C2 overexpressors showed less sensitivity to cytokinin in primary root elongation, lateral root formation, Chl retention and anthocyanin accumulation. Under normal growth conditions ugt76c2 had smaller seeds than the wild type, with accompanying lowered levels of active and N-glucosylated cytokinin forms. The expression levels of cytokinin-related genes such as AHK2, AHK3, ARR1, IPT5 and CKX3 were changed in ugt76c2, suggesting homeostatic control of cytokinin activity. Studies of spatiotemporal expression patterns showed that UGT76C2 was expressed at a relatively higher level in the seedling and developing seed. In their entirety, our data, based mainly on this comparison and opposite phenotypes of knockout and overexpressors, strongly suggest that UGT76C2 is involved in cytokinin homeostasis and cytokinin response in planta through cytokinin N-glucosylation.

Background: Intraoperative magnetic resonance imaging (iMRI) combined with optic radiation neuronavigation may be safer for resection of cerebral lesions involving the optic radiation., Objective: To investigate whether iMRI combined with optic radiation neuronavigation can help maximize tumor resection while protecting the patient's visual field., Methods: Forty-four patients with cerebral tumors adjacent to the optic radiation were enrolled in the study. The reconstructed optic radiations were observed so that a reasonable surgical plan could be developed. During the surgery, microscope-based fiber tract neuronavigation was routinely implemented. The lesion location (lateral or not to the optic radiation) and course of the optic radiation (stretched or not) were categorized, and their relationships to the visual field defect were determined., Results: Analysis of the visible relationship between the optic radiation and the lesion led to a change in surgical approach in 6 patients (14%). The mean tumor residual rate for glioma patients was 5.3% (n = 36) and 0% for patients with nonglioma lesions (n = 8). Intraoperative MRI and fiber tract neuronavigation increased the average size of resection (first and last iMRI scanning, 88.3% vs 95.7%; P < .01). Visual fields after surgery improved in 5 cases (11.4%), exhibited no change in 36 cases (81.8%), and were aggravated in 3 cases (6.8%)., Conclusion: Diffusion tensor imaging information was helpful in surgical planning. When iMRI was combined with fiber tract neuronavigation, the resection rate of brain lesions involving the optic radiation was increased in most patients without harming the patients' visual fields.

The stator design of a new type of spherical motor driven by piezoelectric actuators is developed. A curved piezoelectric actuator is designed to attach to the spherical surface. A series of the curved piezoelectric actuators is laid in a line around a spherical surface. By applying an appropriate voltage signal with phase difference on neighboring actuators, a traveling wave is generated on the hemispherical shell. Each set of curved piezoelectric actuators is designed to provide motion with a single degree-of-freedom (DOF). With two or three sets of the piezoelectric actuators constructed to be mutually perpendicular, the motor can provide 2-DOF or 3-DOF motion. Stator design and analysis and experiment for the 1-, 2-, and 3-DOF conditions are presented in this article. Analytical calculation and experiment results of several fundamental characteristics of the stator are in good agreement. Performance evaluation of rotation speed and torque of the stator and some implementation problems are also addressed.

Background: Eales' disease (ED) is an idiopathic obliterative vasculopathy that usually affects the peripheral retina of young adults. The aim of this study is to investigate Th1/Th2 serum cytokine profiling in patients with ED., Methods: This study included 30 male patients with ED and 10 healthy controls. The ED patients were divided into two subgroups: the vitreous haemorrhage (VH) group (n = 18) and non-vitreous haemorrhage (NVH) group (n = 12). Sixteen patients (six from the VH group and 10 from the NVH group) received glucocorticoid treatment for three months and were followed for six months. Levels of six cytokines including interferon-gamma (IFN-gamma), tumour necrosis factor-alpha (TNF-alpha) and four interleukins (IL-2, IL-4, IL-5 and IL-10) in the serum samples were determined by Luminex assays., Results: Compared to controls, ED patients showed significantly higher levels of IL-10 and TNF-alpha, increased IFN-gamma/IL-4 (Th1/Th2) ratio, and lower levels of IL-4 (p < 0.05). Glucocorticoid treatment caused a restoration in the cytokine levels and the IFN-gamma/IL-4 ratio. Multivariate analysis revealed that reduced IL-4 (less than 4 pg/ml) and elevated IL-10 (greater than 4 pg/ml) levels were independent predictors of ED with odds ratios of 0.024 (95% CI, 0.002-0.255; p = 0.002) and 12.108 (95% CI, 1.045-140.233; p = 0.046), respectively., Conclusion: The findings demonstrate for the first time that there is an imbalance of Th1/Th2 cytokines in ED patients, which can be reversed by glucocorticoid treatment. Additionally, both IL-4 and IL-10 might represent potential diagnostic markers for the disease.

Objective: To probe the intervention and mechanism of the inhibitor of heparanase (PI-88) on the experimental CNV model ., Methods: Experimental CNV was induced by laser photocoagulation in 29 mail (Brown Norway) BN rats (647 nm wave length Krypton laser, 360 mW power, 50 microm spot size, 0.05 second duration), and randomly divided into vacant control group, physiologic saline control group, preventive group and treated group, another 3 rats were acted as normal group, 15 days continuous intraperitoneal injection of PI-88 (25 mg kg(-1) d(-1) was administrated in preventive group (PI-88 was administered the same day as photocoagulation) and treated group (PI-88 was administered 1 week after photocoagulation when CNV had been formed), for the physiologic saline control group, PI-88 was replaced by physiologic saline. To comprehensively evaluate the effect of PI-88 on the CNV by the quantitation of CNV area marked by FITC-dextran in choroid-sclera flat mounts, fluorescence fundus angiography and histopathology; the changes of HPA expression were surveyed by western blot and immunohistochemistry., Results: CNV area of preventive group and treated group decreased 52.1% and 53.8% respectively at the time point of 3 weeks after photocoagulation; the relative thickness of CNV membrane in eyes of treated group had been decreased 46% as that of control group by histopathology; CNV occurrenced 1 week after photocoagulation both in the control group and preventive group, while the fluorescein leakage of the preventive group had been inhibited significantly; 3 weeks after photocoagulation, there had been 7 days discontinuation for the prevent group, the fluorescein leakage did not enhanced, while there had been 15 days continuous administration for the treated group, the fluorescein leakage had been decreased significantly compared to the time point of 2 week; western blot showed that the relative expressed levels of HPA protein in both preventive and treated group decreased; HPA displayed distribution at the leading edge of CNV membrane migrating toward inner retina and the vascular tissue by immunohistochemistry, the expression of HPA was inhibited significantly in treated group., Conclusion: PI-88 may not only prevent CNV in certain degree, but also make it partially subsidize for the existed CNV, and the effect is associated with the inhibition of HPA production.

Objective: To screen and identify the abnormal proteins expressed by hypoxia human retinal pigment epithelium (RPE) in vitro., Methods: It was a experimental study. Protein of normal/hypoxia human RPE cells in exponential phase of growth was extracted, and frozen by liquid nitrogen for proteome study. All samples were performed isoelectric focusing electrophoresis (IEF), separated by isoelectric point (pI); progressed sodium dodecylsulphate polyacrylamide gel electrophoresis (SDS-PAGE) and separated by protein molecular weight (Mr). 2D-gels revealed by coomassie brilliant blue, analyzed by Image Master 2D Elite. Differential proteins spots were screened in normal/hypoxia RPE cell. For identification of differential proteins, peptide masses were analyzed by matrix-assisted laser desorption ionization time-of flight mass spectrometry (MALDI-TOF/MS)., Results: Five hundred and seventy-eight protein spots were obtained in normal group (matching rate within group was 92.90%), and hypoxia group 559 (91.41%). The average matching rate between two groups was 85.47%. There were 32 differential protein spots, which volume value changed > or = 2.0 times. 7 spots were selected randomly for MS, and 5 proteins were identified successfully: HSP70 and HSP60 up-regulated; while beta-actin, beta-tubulin and peroxiredoxin 3 down-regulated., Conclusions: Findings of the in vitro study provide the evidence for expression changes of many proteins in RPE cell with hypoxia state; some of those regulated proteins can result in increasing stress capability obviously in cells, while the major cytoskeletal proteins down-regulated, and the ability of sustained-shapes, keeping order internal structure in cells decreasing. Proteome analysis provides a useful platform in systematic screening of various protein expressions in CNV related diseases.