Comparison of WeatherMax^®and Engame^TMFormulations of Glyphosate on Cotyledon Surface Structure, Chlorophyll A Fluorescence and Shikimate Levels in Isogenic Cotton Cultivars Differing in Roundup Resistance

The effects of Roundup WeatherMax® and EngameTM formulations of glyphosate were investigated on the cotyledons of glyphosate resistant (GR) and glyphosate sensitive (GS) isogenic cotton cultivars. EngameTM is a mixture of glyphosate and 1-aminomethanamide dihydrogen tetraoxosulfate (AMADS). Fully expanded cotton cotyledons treated with EngameTM or AMADS developed surface lesions within 2 hours after treatment whereas surfactant-treated control or WeatherMax®-treated tissues did not develop lesions. The EngameTM and AMADS damage appeared as depressions which were confirmed by scanning electron microscopy. Light micrographs of cross sections through the depressions revealed collapsed and compressed epidermal and mesophyll cells with congealed cytoplasmic contents in the palisade and spongy mesophyll cells. Changes to photosynthetic electron transport were evident at 4 hours after treatment (HAT) in all treatments as revealed by chlorophyll A fluorescence. In GR cotton, the fluorescence perturbations decreased with time such that at 72 HAT EngameTM-treated cotyledons could not be distinguished from the surfactant-or WeatherMax®-treated plants. The GS cotton continued to show progressive decreases in the fluorescence parameters Fv/Fm and performance index (PI) to 72 HAT. Shikimate levels increased following glyphosate treatment in glyphosate sensitive cotton and EngameTM caused a two-to three-fold greater increase in shikimate compared to WeatherMax®. These results indicate that the EngameTM-based glyphosate formulation involved structural tissue damage which likely increased glyphosate uptake and subsequently increased inhibition of photosynthesis and the shikimate pathway.

Overexpression of the Wounding-Responsive Gene AtMYB15 Activates the Shikimate Pathway in Arabidopsis

The MYB transcription factor genes play important roles in many developmental processes and various defense responses of plants. The shikimate pathway is a major biosynthetic pathway for the production of three aromatic amino acids and other aromatic compounds that are involved in multiple responses of plants, including protection against UV and defense. Herein, we describe the characterization of the R2R3-MYB gene AtMYB15as an activator of the shikimate pathway in Arabidopsis. The AtMYB15 protein is nuclear localized and a transcriptional activation domain is found in its C-terminal portion. Northern blots showed that AtMYB15 is an early wounding-inducible gene. Resutls of microarray analysis, confirmed using quantitative real-time polymerase chain reaction, showed that overexpression of AtMYB15 in transgenic plants resulted in elevated expression of almost all the genes involved in the shikimate pathway. Bioinformatics analysis showed that one or more AtMYB15-binding AC elements were detected in the promoters of these upregulated genes. Furthermore, these genes in the shikimate pathway were also found to be induced by wounding. These data suggest an important role of AtMYB15as a possible direct regulator of the Arabidopsis shikimate pathway in response to wounding.

The shikimate pathway regulates programmed cell death

Programmed cell death(PCD)is essential for both plant development and stress responses including immunity.However,how plants control PCD is not well-understood.The shikimate pathway is one of the most important metabolic pathways in plants,but its relationship to PCD is unknown.Here,we show that the shikimate pathway promotes PCD in Arabidopsis.We identify a photoperiod-dependent lesion-mimic mutant named Lesion in short-day(lis),which forms spontaneous lesions in short-day conditions.Mapbased cloning and whole-genome resequencing reveal that LIS encodes MEE32,a bifunctional enzyme in the shikimate pathway.Metabolic analysis shows that the level of shikimate is dramatically increased in lis.Through genetic screenings,three suppressors of lis(slis)are identified and the causal genes are cloned.SLISes encode proteins upstream of MEE32 in the shikimate pathway.Furthermore,exogenous shikimate treatment causes PCD.Our study uncovers a link between the shikimate pathway and PCD,and suggests that the accumulation of shikimate is an alternative explanation for the action of glyphosate,the most successful herbicide.

Shikimic acid accelerates phase change and flowering in Chinese jujube

The juvenile-to-adult phase change with first flowering as the indicator plays a crucial role in the lifecycle of fruit trees. However, the molecular mechanisms underlying phase change in fruit trees remain largely unknown. Shikimic acid (ShA) pathway is a main metabolic pathway closely related to the synthesis of hormones and many important secondary metabolites participating in plant phase change. So,whether ShA regulates phase change in plants is worth clarifying. Here, the distinct morphological characteristics and the underlying mechanisms of phase change in jujube (Ziziphus jujuba Mill.), an important fruit tree native to China with nutritious fruit and outstanding tolerance abiotic stresses, were clarified. A combined transcriptome and metabolome analysis found that ShA is positively involved in jujube(Yuhong’×Xing 16’) phase change. The genes in the upstream of ShA synthesis pathway (ZjDAHPS, ZjDHQS and ZjSDH), the contents of ShA and the downstream secondary metabolites like phenols were significantly upregulated in the phase change period. Further, the treatment of spraying exogenous ShA verified that ShA at a very low concentration (60 mg·L^(-1)) can substantially speed up the phase change and flowering of jujube and other tested plants including Arabidopsis, tomato and wheat. The exogenous ShA (60 mg·L^(-1)) treatment in jujube seedlings could increase the accumulation of endogenous ShA, enhance leaf photosynthesis and the synthesis of phenols especially flavonoids and phenolic acids, and promote the expression of genes (ZjCOs, ZjNFYs and ZjPHYs) involved in flowering pathway. Basing on above results, we put forward a propose for the underlying mechanism of ShA regulating phase change, and a hypothesis that ShA could be considered a phytohormone-like substance because it is endogenous, ubiquitous, movable and highly efficient at very low concentrations. This study highlights the critical role of ShA in plant phase change and its phytohormone-like properties.

Novel and Efficient Syntheses of Four Useful Shikimate- derived Epoxy Chiral Building Blocks via Cyclic Sulfite Intermediates

All of the four stereoisomers of methyl 4,5-epoxy-3-hydroxy-cyclohex-l-ene-carboxylate （la--ld） are useful chiral building blocks. Novel and efficient syntheses of these four epoxy chiral building blocks from naturally abun- dant （--）-shikimic acid （2） via cyclic sulfite intermediates are described in this article. The targeted compound （3R,4R,5S）-la was synthesized via four steps from （--）-shikimic acid in 79% overall yield. The other three targeted compounds （3S,4R,5S）-lb, （3S,4S,5R）-1c and （3R,4S,5R）-ld were synthesized via seven steps from （--）-shikimic acid in 56%, 64% and 65% overall yields, respectively.

Natural sources, biosynthesis, biological functions, and molecular mechanisms of shikimic acid and its derivatives

Shikimic acid is a hydroaromatic compound possessing critical biological properties,such as antibacterial and antiviral activity.This review mainly focused on shikimic acid and its derivatives.We first briefly introduced the sources of shikimic acid and its derivatives and discussed their biosynthesis.Several in vitro and in vivo studies indicate that shikimic acid and its derivatives exhibit diverse bioactivities,such as antioxidant,antiviral,anti-inflammatory,antibacterial,hypolipidemic,bone protective,skin protective,neuroprotective,and antidiabetic activities.We mainly focused on the related molecular mechanisms.Overall,the wide range of bioactivities of shikimic acid and its derivatives indicate that a more detailed exploration of their potential for the prevention and treatment of certain diseases is warranted.

Changes of chlorogenic acid content and its synthesis-associated genes expression in Xuehua pear fruit during development

According to synthetic pathway of plant chlorogenic acid （CGA）, the expression patterns of genes encoding enzymes that are associated with CGA synthesis were studied in normally developed Xuehua pear fruit. The study demonstrated that CGA content in peel and flesh of Xuehua pear decreased as fruit development progressed, with a higher level in peel. The expression levels of PbPAL 1, PbPAL2, PbC3H, PbC4H, Pb4CL 1, Pb4CL2, Pb4CL6, PbHC T1 and PbHC T3 genes decreased in fruit, which was consistent with the pattern of variation in CGA content. That indicated that these genes might be key genes for influencing fruit CGA synthesis in Xuehua pear. However, Pb4CL7 gene expression profile is not consistent with variation of CGA content, hence, it may not be a key gene involved in CGA synthesis.

月见草中一个新的莽草酸衍生物

为了研究月见草Oenothera biennis L.的化学成分,采用硅胶、Sephadex LH-20凝胶、MCI gel CHP-20等柱色谱并结合半制备高效液相等分离技术,利用现代波谱学手段进行化合物结构鉴定。从月见草50%丙酮提取物中分离得到10个化合物,分别鉴定为dimethyl 3-lactyl shikimate(1)、methyl shikimate(2)、3,3′,4-O-三甲基鞣花酸(3)、3,3′-二甲氧基-鞣花酸(4)、3,4,3′-三甲氧基鞣花酸(5)、alternariol 9-methyl ether(6)、2,3-二羟基-1-(4-羟基-3,5-二甲氧基苯基)-1-丙酮(7)、scirpyrone H(8)、dimethoxydimethylphthalide(9)、clearanol C(10)。其中化合物1为新化合物,化合物2~10均为首次从该植物中分离得到。化合物2和3可显著提高TGF-β1诱导BEAS-2B细胞的活力,推测其具有潜在的肺保护作用。

Facile Syntheses of (6S)-6-Fluoroshikimic Acid and (6R)-6-Hydroxyshikimic Acid

(6S)-6-Fluoroshikimic acid 2 and (6 R ) 6 hydroxyshikimic acid 3 have been synthesized via an OsO 4 catalysed dihydroxylation of diene 7, which was derived from (-) shikimic acid 1.

Occurrence of Glyphosate-Resistant Horseweed (Conyza canadensis) Population in China

Horseweed （Conyza canadensis）, an invasive alien weed, is one of the main weeds in orchards in China. Although glyphosate has been used for control of horseweed and many other weeds in orchards for more than 25 years in China, a case of glyphosate-resistant horseweed has not been identified in orchard in China so far despite glyphosate-resistant horseweed cases have been reported in some other countries. Seeds of 25 horseweed populations were collected from different orchards with different glyphosate application history. Potted seedlings with 11-13-leaf growth stage were treated with glyphosate at 0.035, 0.07, 0.14, 0.28, 0.56, 1.12, 2.24, 4.48, and 8.96 kg a.i. ha-1. The dosage dependence response curve of each population was constructed with Log-logistic dose response regression equations. The ED50 value of each population was calculated and compared with the susceptible population from China. Different populations had different relative glyphosate-resistant levels which increased with the number of years of glyphosate application. Two populations with the highest resistance levels, 8.28 and 7.95 times, were found in Ningbo, Zhejiang Province, China, where glyphosate was used for weed control in orchards twice each year for 15 yr. The two resistant populations accumulated approximately two to four times less shikimic acid than the two susceptible populations 48 h after glyphosate application.

Glyphosate Effects on Sugarcane Metabolism and Growth

Glyphosate is the most widely used herbicide in the world. In sugarcane, it is used as a herbicide when applied at its field rate, but it is also used as ripener when applied as low doses. However, the effects of glyphosate on plant metabolism and sugarcane growth are not fully understood. This study aimed to evaluate the metabolic changes and the effects on sugarcane plant growth caused by the application of different doses of glyphosate. Sugarcane plants were grown in a greenhouse and subjected to glyphosate applications at doses of 7.2;18;36;72;180;360 and 720 g a.e. ha-1. Plants grown without an herbicide application were used as a control. Plants from each treatment were collected at 2, 7, 14, and 21 days after treatment (DAT) application to quantify the levels of shikimic acid, quinic acid, shikimate-3-phosphate, glyphosate, α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA), phenylalanine, tyrosine, and tryptophan. Visual evaluations of plant intoxication were performed at the same time as the collection of plants, and the quantification of their shoot dry biomass was performed at 21 DAT. At doses of glyphosate greater than 72 g a.e. ha-1, increases in the levels of shikimic acid, quinic acid, and shikimate-3-phosphate occurred and AMPA was detected in the plants. Initially, glyphosate caused increases in the plant levels of phenylalanine and tyrosine at doses of 72 and 180 g a.e. ha-1, although a decrease in the levels of aromatic amino acids subsequently occurred at and above the doses of 72 or 180 g a.e. ha-1. The doses ranging from 7.2 to 36 g a.e. ha-1 promoted an increase in plant shoot biomass, and doses greater than 72 g a.e. ha-1 caused significant reductions in dry mass.

Enantioselective Total Synthesis of the (+) Antipode of Zeylenone

Starting from shikimic acid, the total synthesis of zeylenone was studied. The productwas proved to be the (+)antipode of zeylenone through analysis and comparison of their respectivespectra (including NMR, MS, IR and CD) and optical data. The absolute configuration of thenatural product was thus determined to be (1S,2S,3R).

Shikimic Acid Promotes Oligodendrocyte Precursor Cell Differentiation and Accelerates Remyelination in Mice

The obstacle to successful remyelination in demyelinating diseases, such as multiple sclerosis, mainly lies in the inability of oligodendrocyte precursor cells(OPCs) to differentiate, since OPCs and oligodendrocytelineage cells that are unable to fully differentiate are found in the areas of demyelination. Thus, promoting the differentiation of OPCs is vital for the treatment of demyelinating diseases. Shikimic acid(SA) is mainly derived from star anise, and is reported to have antiinfluenza, anti-oxidation, and anti-tumor effects. In the present study, we found that SA significantly promoted the differentiation of cultured rat OPCs without affecting their proliferation and apoptosis. In mice, SA exerted therapeutic effects on experimental autoimmune encephalomyelitis(EAE), such as alleviating clinical EAE scores, inhibiting inflammation, and reducing demyelination in the CNS. SA also promoted the differentiation of OPCs as well as their remyelination after lysolecithin-induced demyelination.Furthermore, we showed that the promotion effect of SA on OPC differentiation was associated with the up-regulation of phosphorylated m TOR. Taken together, our resultsdemonstrated that SA could act as a potential drug candidate for the treatment of demyelinating diseases.

Enhanced production of shikimic acid using a multi-gene co-expression system in Escherichia coli

Shikimic acid(SA) is the key synthetic material for the chemical synthesis of Oseltamivir, which is prescribed as the front-line treatment for serious cases of influenza. Multi-gene expression vector can be used for expressing the plurality of the genes in one plasmid, so it is widely applied to increase the yield of metabolites. In the present study, on the basis of a shikimate kinase genetic defect strain Escherichia coli BL21(?aro L/aro K, DE3), the key enzyme genes aro G, aro B, tkt A and aro E of SA pathway were co-expressed and compared systematically by constructing a series of multi-gene expression vectors. The results showed that different gene co-expression combinations(two, three or four genes) or gene orders had different effects on the production of SA. SA production of the recombinant BL21-GBAE reached to 886.38 mg·L^(-1), which was 17-fold(P < 0.05) of the parent strain BL21(?aro L/aro K, DE3).

Improved Stereoselective Syntheses of （-t-）-Valiolamine and （-I-）-Valienamine Starting from （-）-Shikimic Acid

Improved stereoselective syntheses of the targeted compounds （＋）-valiolamine 1 and （＋）-valienamine 2 starting from naturally abundant （–）-shikimic acid are described. A common key intermediate compound 7 was first synthesized from （–）-shikimic acid in 9 steps. The compound 7 was then converted to （＋）-valiolamine 1 in 3 steps, and was also converted to （＋）-valienamine 2 in 4 steps. In summary, （＋）-valiolamine 1 and （＋）-valienamine 2 were synthesized from （–）-shikimic acid in 12 （or 13） steps in 40% and 39% overall yields, respectively.