ABSCISIC ACID-INSENSITIVE 5-KIP-RELATED PROTEIN 1-SHOOT MERISTEMLESS modulates reproductive development of Arabidopsis.

Soil (or plant) water deficit accelerates plant reproduction. However, the underpinning molecular mechanisms remain unknown. By modulating cell division/number, ABSCISIC ACID-INSENSITIVE 5 (ABI5), a key bZIP (basic (region) leucine zippers) transcription factor, regulates both seed development and abiotic stress responses. The KIP-RELATED PROTEIN (KRP) cyclin-dependent kinases (CDKs) play an essential role in controlling cell division, and SHOOT MERISTEMLESS (STM) plays a key role in the specification of flower meristem identity. Here, our findings show that abscisic acid (ABA) signaling and/or metabolism in adjust reproductive outputs (such as rosette leaf number and open flower number) under water-deficient conditions in Arabidopsis (Arabidopsis thaliana) plants. Reproductive outputs increased under water-sufficient conditions but decreased under water-deficient conditions in the ABA signaling/metabolism mutants abscisic acid2-1 (aba2-1), aba2-11, abscisic acid insensitive3-1 (abi3-1), abi4-1, abi5-7, and abi5-8. Further, under water-deficient conditions, ABA induced-ABI5 directly bound to the promoter of KRP1, which encodes a CDK that plays an essential role in controlling cell division, and this binding subsequently activated KRP1 expression. In turn, KRP1 physically interacted with STM, which functions in the specification of flower meristem identity, promoting STM degradation. We further demonstrate that reproductive outputs are adjusted by the ABI5-KRP1-STM molecular module under water-deficient conditions. Together, our findings reveal the molecular mechanism by which ABA signaling and/or metabolism regulate reproductive development under water-deficient conditions. These findings provide insights that may help guide crop yield improvement under water deficiency.

Feedback loop promotes sucrose accumulation in cotyledons to facilitate sugar-ethylene signaling-mediated, etiolated-seedling greening.

De-etiolation is indispensable for seedling survival and development. However, how sugars regulate de-etiolation and how sugars induce ethylene (ET) for seedlings to grow out of soil remain elusive. Here, we reveal how a sucrose (Suc) feedback loop promotes de-etiolation by inducing ET biosynthesis. Under darkness, Suc in germinating seeds preferentially induces 1-amino-cyclopropane-1-carboxylate synthase (ACS7; encoding a key ET biosynthesis enzyme) and associated ET biosynthesis, thereby activating ET core component ETHYLENE-INSENSITIVE3 (EIN3). Activated EIN3 directly inhibits the function of Suc transporter 2 (SUC2; a major Suc transporter) to block Suc export from cotyledons and thereby elevate Suc accumulation of cotyledons to induce ET. Under light, ET-activated EIN3 directly inhibits the function of phytochrome A (phyA; a de-etiolation inhibitor) to promote de-etiolation. We therefore propose that under darkness, the Suc feedback loop (Suc-ACS7-EIN3-|SUC2-Suc) promotes Suc accumulation in cotyledons to guarantee ET biosynthesis, facilitate de-etiolation, and enable seedlings to grow out of soil.

Phaeotremella camelliae sp. nov. (Phaeotremellaceae, Tremellales), A Novel Yeasts Isolated from Tea-Oil Fruits in Jiangxi Province, China.

Sixty-two isolates among the 65 yeast strains isolated from Jiangxi province, China, were identified into 15 known species based on the sequence analysis of the D1/D2 domains of the LSU rRNA and ITS region. The other three strains, GaoanZ14, GaoanC57, and GaoanC191, isolated from tea-oil fruits, were identified as two undescribed species of Phaeotremella based on the multiple gene sequence analysis, physiological, and biochemical comparisons. Strains GaoanC57 and GaoanC191 had one substitution difference both in the D1/D2 domains of the LSU rRNA and ITS region. They formed a separate branch from the other Phaeotremella species in the D1/D2 and multiple genes trees, and differed from the known species by at least 10 nucleotide substitutions in the D1/D2 domains and more than 6% mismatches in the ITS region. The phylogenetic analysis indicated that those two strains represent a novel species of Phaeotremella, for which the names Phaeotremella camelliae sp. nov. (Holotype CGMCC 2.6141, Mycobank MB832699) is proposed. Only one strain, GaoanZ14, represents the other undescribed species of Phaeotremella, so it will be described in latter when more strains are found.

[Percutaneous transforaminal endoscopic discectomy via lateral position assisted with local anesthesia for lumbar disc herniation in elder with systemic diseases].

OBJECTIVE: To explore the clinical effect of percutaneous transforaminal endoscopic discectomy via lateral position assisted with local anesthesia for lumbar disc herniation in elder with systemic diseases. METHODS: The clinical data of 44 elder patients with lumbar disc herniation and systemic diseases treated from June 2013 to June 2017 were retrospectively analyzed. Including 26 males and 18 females, aged 70 to 86 years old with an average of(77.5±3.5) years, course of disease was 3 weeks to 6 months. Percutaneous transforaminal endoscopic discectomy via lateral position assisted with local anesthesia was performed in the patients. Postoperative straight leg-raising degree change and Nakai criteria were used to evaluate the operative effect. Visual analogue scale(VAS) and JOA scoring system were used to compare the symptom improvement between preoperative and postoperative. RESULTS: No serious complications occurred during and after the operation, and the coexisting medical diseases were not aggravated. All patients safely survived the perioperative period. All the 44 cases were followed up from 6 to 30 months with an average of 20 months. Postoperative straight leg raising angle was obviously improved. At final follow-up, according to Nakai standard, 41 cases got excellent results, 3 good. VAS scores were significantly decreased and JOA scores were significantly increased in final follow-up(P<0.01). CONCLUSIONS: Percutaneous transforaminal endoscopic discectomy via lateral position assisted with local anesthesia is a safe, effective, less invasive technique for the treatment of lumbar disc herniation in elder with systemic diseases. Multidisciplinary collaboration and perioperative well controlled disease are essential for early rehabilitation of such patients.

Sucrose Signaling Regulates Anthocyanin Biosynthesis Through a MAPK Cascade in Arabidopsis thaliana.

Anthocyanin accumulation specifically depends on sucrose (Suc) signaling. However, the molecular basis of this process remains unknown. In this study, in vitro pull-down assays identified ETHYLENE-INSENSITIVE3 (EIN3), a component of both sugar signaling or/and metabolism. This protein interacted with YDA, and the physiological relevance of this interaction was confirmed by in planta co-immunoprecipitation, yeast two-hybrid (Y2H) assay, and bimolecular fluorescence complementation. Ethylene insensitive3-like 1 (eil1) ein3 double-mutant seedlings, but not ein3-1 seedlings, showed anthocyanin accumulation. Furthermore, ein3-1 suppressed anthocyanin accumulation in yda-1 plants. Thus, EMB71/YDA-EIN3-EIL1 may form a sugar-mediated gene cascade integral to the regulation of anthocyanin accumulation. Moreover, the EMB71/YDA-EIN3-EIL1 gene cascade module directly targeted the promoter of Transparent Testa 8 (TT8) by direct EIN3 binding. Collectively, our data inferred a molecular model where the signaling cascade of the YDA-EIN3-TT8 appeared to target TT8 via EIN3, thereby modulating Suc signaling-mediated anthocyanin accumulation.

Purification and identification of an actinomycin D analogue from actinomycetes associated with Ganoderma applanatum via magnetic molecularly imprinted polymers and tandem mass spectrometry.

Actinomycetes are main producers of antibiotics and targeted screening could improve the efficiency of discovering new drugs. This study describes, for the first time, the isolation of endophytic actinomycetes from the macrofungus Ganoderma applanatum. To increase the efficiency of screening, novel actinomycin D (Act D) molecularly-imprinted polymers were adsorbed to the surface of Fe3O4@SiO2 magnetic microspheres (MMIPs) and using in the isolation. A monolithic column prepared with magnetic molecularly imprinted polymers was employed to adsorb actinomycin D and its analogues for selective analysis and identification via MS/MS spectroscopy. The MMIP-monolithic column was selective for the structural features of Act D and its analogue, and the maximum loading of the MMIPs for Act D was ∼23.5 µg/g. The recognition time of the Act D was 20-30 min and had good discriminative ability. A new analogue was identified from endophytic actinomycetes KLBMP 2541, and it was purified using MMIPs comparison with MMIPs-solid phase extraction. Structural identification analysis confirmed that the new analogue was 2-methyl-actinomycin D, which has better anti-tumor activity than Act D. The presented method combines the advantages of MMIPs and MS with popular solutions to enable high affinity and selectivity screening of specific antibiotics from endophytic actinomycetes.

Nocardia rhizosphaerihabitans sp. nov., a novel actinomycete isolated from a coastal soil.

An actinomycete strain, designated KLBMP S0039T, was isolated from the rhizosphere soil of Lycium Linn., collected from the coastal region in Lianyungang, Jiangsu Province, eastern PR China, and was studied to determine its taxonomic position. The isolate showed a combination of morphological and chemotaxonomic properties typical of the members of the genus Nocardia. The cell wall contained meso-diaminopimelic acid as the diagnostic diamino acid and the whole-cell sugars were galactose, arabinose, glucose and ribose. The predominant menaquinone was identified as MK-8(H4ω-cycl). The diagnostic phospholipids were found to be diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol and unknown lipids. The predominant cellular fatty acids were identified as C16 : 0, C18 : 0, C18 : 1ω9c, and 10-methyl C18 : 0 [tuberculostearic acid (TBSA)]. The G+C content of the genomic DNA was determined to be 68.2 mol%. The 16S rRNA gene sequence similarity indicated that KLBMP S0039T was most closely related to Nocardia neocaledoniensis NBRC 108232T (99.4 % 16S rRNA gene sequence similarity) and Nocardia asteroides NBRC 15531T (99.2 %), similarities to other type strains of species of the genus Nocardia were found to be less than 98.6 %. However, DNA-DNA relatedness values and phenotypic data indicated that KLBMP S0039T could be clearly distinguished from the closely related species of the genus Nocardia. On the basis of polyphasic taxonomic data, it is concluded that KLBMP S0039T represents a novel species of the genus Nocardia, for which the name Nocardiarhizosphaerihabitans sp. nov. is proposed. The type strain is KLBMP S0039T (=KCTC 39693T=CGMCC 4.7329T).

Complete genome sequence of endophyte Bacillus flexus KLBMP 4941 reveals its plant growth promotion mechanism and genetic basis for salt tolerance.

Bacillus flexus KLBMP 4941 is a halotolerant endophyte isolated from the halophyte Limonium sinense. This strain can improve host seedling growth under salt stress conditions. We here report the complete genome information of endophyte KLBMP 4941. It has a circular chromosome and two plasmids for a total genome 4,104,242 bp in size with a G+C content of 38.09%. Genes related to plant growth promotion (PGP), such as those associated with nitrogen fixation, siderophore, spermidine, and acetoin synthesis were found in the KLBMP 4941 genome. Some genes responsible for high salinity tolerance, like genes associated with the Na+/H+ antiporter, glycine betaine transporter, and betaine-aldehyde dehydrogenase were also found in the KLBMP 4941 genome. The genome analysis will provide better understanding of the mechanisms underlying the promotion of plant growth in strain KLBMP 4941 under salt stress conditions and its ability to adapt to coastal salt marsh habitats, and provide a basis for its further biotechnological applications in agriculture.

Corrigendum: The Arabidopsis ANGUSTIFOLIA3-YODA Gene Cascade Induces Anthocyanin Accumulation by Regulating Sucrose Levels.

[This corrects the article on p. 1728 in vol. 7, PMID: 27920784.].

Corrigendum: Seed Embryo Development Is Regulated via an AN3-MINI3 Gene Cascade.

[This corrects the article on p. 1645 in vol. 7, PMID: 27857719.].

Soluble Sugar Accumulation Can Influence Seed Size via AN3-YDA Gene Cascade.

In higher plants, seed size is central to many aspects in evolutionary fitness and is a crucial agricultural trait. In this study, Arabidopsis an3 (angustifolia3) mutants present with increased seed size. Target-gene analysis revealed that YDA, which encodes a mitogen-activated protein kinase kinase kinase, is a target gene of AN3. Indeed, the loss of YDA function decreases seed size. Furthermore, AN3 and YDA mutations both disrupt normal sucrose and glucose contents and cause altered seed size in an3 or yda mutants. With these results, we provide a molecular model in which soluble sugar accumulation might affect seed size regulation via the AN3-YDA gene cascade. Our findings guide the synthesis of a model that predicts the integration of soluble sugar accumulation at AN3 to control the establishment of seed size.

Glutamicibacter halophytocola sp. nov., an endophytic actinomycete isolated from the roots of a coastal halophyte, Limonium sinense.

A novel actinobacterium, designated KLBMP 5180T, was isolated from the surface-sterilized root of a coastal halophyte, Limonium sinense, collected from the city of Lianyungang, Jiangsu Province, eastern China. The isolate was Gram-stain-positive, aerobic and non-motile. The components of the cell-wall peptidoglycan were lysine, glutamic acid and alanine. The predominant menaquinone was MK-9. The polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol, one unknown phospholipid, one unidentified glycolipid and two unidentified lipids. anteiso-C15 : 0 and iso-C16 : 0 were the major cellular fatty acids. The DNA G+C content of strain KLBMP 5180T was 60.0 mol%. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain KLBMP 5180T belongs to the genus Glutamicibacter and was related most closely to Glutamicibacter nicotianae DSM 20123T (99.3 % similarity), Glutamicibacterarilaitensis Re117T (99.3 %) and Glutamicibacter mysorens LMG 16219T (99.1 %); similarity to other type strains of the genus Glutamicibacter was lower than 98.5 %. However, DNA-DNA relatedness values between strain KLBMP 5180T, G . nicotianae DSM 20123T, G. arilaitensis Re117T and G. mysorens LMG 16219T were 47.5±2.6, 51.3±3.1 and 41.2±4.3 %, respectively. The combination of DNA-DNA hybridization, phylogenetic, phenotypic and chemotaxonomic data supported the suggestion that strain KLBMP 5180T represents a novel species of the genus Glutamicibacter, for which the name Glutamicibacterhalophytocola sp. nov. is proposed. The type strain is KLBMP 5180T (=DSM 101718T=KCTC 39692T).

Selenylation of Polysaccharide from the Sweet Potato and Evaluation of Antioxidant, Antitumor, and Antidiabetic Activities.

Interest in sweet potato as a functional food is growing. A polysaccharide (SWP) was isolated from the sweet potato tuber and elucidation of its structure as composed of rhamnose, glucose, and galactose undertaken. To improve its activity, selenylation of this novel polysaccharide (Se-SWP) was undertaken by using microwave synthesis. In vitro evaluation showed that the Se-SWP has excellent antioxidant activity on scavenging free radicals and reducing capacity. In vivo antitumor evaluation showed selenylation polysaccharide could effectively inhibit tumor growth (>50%) and adjust immune factor levels in the mice (IL-2, TNF-α, and VEGF). The antidiabetic potential of Se-SWP was tested in STZ-induced diabetic rats. The results indicated that the Se-SWP treatment significantly reduced the levels of malondialdehyde and other disadvantageous factors that were increased by the STZ treatment. Meanwhile, the Se-SWP treatment caused a significant increase in the activities of enzymatic antioxidants and the levels of nonenzymatic antioxidants in the organs of diabetic rats. All of the activity evaluations indicated that the selenylation method could improve the activity of sweet potato polysaccharide and its efficacy as a potential therapeutic, which will be the focus of further study.

KB(PO4)F: a novel acentric deep-ultraviolet material.

Two challenges to grow KBe2BO3F2 (KBBF), the best known deep-ultraviolet nonlinear optical (NLO) material to date, are the limited crystal sizes and the use of a highly toxic element (Be). Herein we report on the discovery of a novel anhydrous non-centrosymmetric alkali fluorinated borophosphate KB(PO4)F (KBPF) featuring a cut-off wavelength of less than 200 nm and a large second-harmonic generation (SHG) effect similar to KH2PO4 (KDP), hence representing a new promising deep-ultraviolet NLO material. The KBPF crystals consisting of common elements can be grown using green and cost effective processes and do not show any detectable hygroscopicity. The title compound also features a 2-dimensional layer [BPO4F]∞ built from [BO3F]4- and [PO4]3- tetrahedral groups but has much stronger interlayer bonds than KBBF, allowing the growth of large crystals. The title compound has been characterized by PXRD, SEM, TG-DSC, FTIR, UV-Vis-NIR diffuse reflectance and SHG analyses as well as single-crystal X-ray structure refinements. The optical properties of KBPF have also been evaluated by first-principles calculations at the density functional theory (DFT) level.

The Arabidopsis ANGUSTIFOLIA3-YODA Gene Cascade Induces Anthocyanin Accumulation by Regulating Sucrose Levels.

Anthocyanin accumulation specifically depends on sucrose (Suc) signaling/levels. However, the gene cascades specifically involved in the Suc signaling/level-mediated anthocyanin biosynthetic pathway are still unknown. Arabidopsis ANGUSTIFOLIA3 (AN3), a transcription coactivator, is involved in the regulation of leaf shape and drought tolerance. Recently, an AN3-CONSTITUTIVE PHOTOMORPHOGENIC 1 gene cascade has been reported to regulate the light signaling-mediated anthocyanin accumulation. Target gene analysis indicates that AN3 is associated with the YODA (YDA) promoter, a mitogen-activated protein kinase kinase kinase, in vivo for inducing anthocyanin accumulation. Indeed, loss-of-function mutants of YDA showed significantly increased anthocyanin accumulation. YDA mutation can also suppress the decrease in an3-4 anthocyanin accumulation. Further analysis indicates that the mutations of AN3 and YDA disrupt the normal Suc levels because of the changes of invertase activity in mutants of an3 or yda, which in turn induces the alterations of anthocyanin accumulation in mutants of an3 or yda via unknown regulatory mechanisms.

Seed Embryo Development Is Regulated via an AN3-MINI3 Gene Cascade.

In agriculture, seed mass is one of the most important components related to seed yield. MINISEED3 (MINI3) which encodes the transcriptional activator WRKY10, is thought to be a pivotal regulator of seed mass. In Arabidopsis SHORT HYPOCOTYL UNDER BLUE1 (SHB1) associates with the promoter of MINI3, regulating embryo cell proliferation (both cell division and elongation), which, in turn, modulates seed mass. Furthermore, the recruitment of SHB1 via MINI3 to both its cognate promoter and that of IKU2 implies a two-step amplification for countering the low expression level of IKU2, which is thought to function as a molecular switch for seed cavity enlargement. However, it is largely unknown how embryo cell proliferation, which encompasses both cell division and elongation, is regulated by SHB1 and MINI3 function. Here, we show that a loss of function mutation within the transcriptional coactivator ANGUSTIFOLIA3 (AN3), increases seed mass. Further, AN3 associates with the MINI3 promoter in vivo. Genetic evidence indicates that the absence of MINI3 function suppresses the decrease of cell number observed in an3-4 mutants by regulating cell division and in turn inhibits increased cell size of the an3-4 line by controlling cell elongation. Thus, seed embryo development is modulated via an AN3-MINI3 gene cascade. This regulatory model provides a deeper understanding of seed mass regulation, which may in turn lead to increased crop yields.

A novel recycling system for nano-magnetic molecular imprinting immobilised cellulases: Synergistic recovery of anthocyanin from fruit and vegetable waste.

Fruit and vegetable waste (FVW) is become a serious problem in developing countries. Enzymolysis is a potentially useful method for the treatment of FVW. In the present study, novel recycled magnetic molecular imprinting immobilised cellulases were prepared based on magnetic modified chitosan (MCTS) and Fe3O4. The properties of obtained were characterised by IR and grain-size measurements. Evaluation of a single factor affecting the loading efficiency of supports and the mixed immobilised enzymes showed better capacity than single immobilised, or free, enzymes. The immobilisation process could improve cellulase stability and repeatability of the method. Meanwhile, the kinetic parameters were also verified. The immobilised enzymes retained most of their capacity after 60days' storage while free enzymes lost it within 30days. Tests showed that the immobilised enzymes developed excellent capacity and five anthocyanins were collected.

Essential Oil from Sweet Potato Vines, a Potential New Natural Preservative, and an Antioxidant on Sweet Potato Tubers: Assessment of the Activity and the Constitution.

Pathogenic fungi and oxidation are the major factors that cause the deterioration of sweet potatoes and also cause the loss of quality that makes consumption unsafe. In the present study, the in vitro results demonstrate that the essential oil from sweet potato vines exhibits significantly enhanced activity compared to that of the control. Furthermore, the essential oil can actively inhibit the growth of some common microorganisms inducing pathogenic bacteria and fungi (inhibition rates above 50% at low concentrations). A total of 31 constituents were identified using GC-MS and confirmed that linalool and p-hydroxybenzoic acid are the major active ingredients. The experiment involving actual tubers showed that the essential oil could retains its quality and effectiveness again the fungus disease. This suggests that it could be used in the food industry to increase the shelf life of stored produce (tubers) to ensure food safety without the use of additives or preservatives.

Cloning and Characterization of a Squalene Synthase Gene from the Chaga Medicinal Mushroom, Inonotus obliquus (Agaricomycetes).

Squalene synthase catalyzes the condensation of 2 molecules of farnesyl diphosphate to produce squalene, the first committed precursor for sterol, brassinosteroid, and triterpene biosynthesis. A squalene synthase gene, designated IoSQS, was isolated from Inonotus obliquus, a medicinal mushroom that produces a plethora of bioactive triterpenes. IoSQS complementary DNA was found to contain an open reading frame of 1476 bp, encoding a protein of 491 amino acids with a calculated molecular mass of 55.85 kDa. The IoSQS genomic DNA sequence consisted of 1813 bp and contained 4 exons and 3 introns. The restriction fragment polymorphisms revealed by Southern blot analysis suggested that IoSQS was a single-copy gene. Promoter analysis indicated that the 5' upstream region of IoSQS possessed various potential elements associated with physiological and environmental factors. The expression pattern of IoSQS in different stages and under methyl jasmonate treatment correlated with the accumulation of total triterpenoids and was consistent with the predicted results of the IoSQS promoter region. The N-terminal 466 residues of the hydrophilic sequence were expressed as a His-tagged protein in Escherichia coli, and the resultant bacterial crude extract was incubated with farnesyl diphosphate and NADPH. Squalene was detected in vitro in reaction mixture by high-performance liquid chromatography analysis. These results suggest that the IoSQS enzyme is involved in squalene production in I. obliquus.