GhKWL1 Upregulates GhERF105 but Its Function Is Impaired by Binding with VdISC1, a Pathogenic Effector of Verticillium dahliae

Verticillium wilt, caused by Verticillium dahliae, is a devastating disease for many important crops, including cotton. Kiwellins (KWLs), a group of cysteine-rich proteins synthesized in many plants, have been shown to be involved in response to various phytopathogens. To evaluate genes for their function in resistance to Verticillium wilt, we investigated KWL homologs in cotton. Thirty-five KWL genes (GhKWLs) were identified from the genome of upland cotton (Gossypium hirsutum). Among them, GhKWL1 was shown to be localized in nucleus and cytosol, and its gene expression is induced by the infection of V. dahliae. We revealed that GhKWL1 was a positive regulator of GhERF105. Silencing of GhKWL1 resulted in a decrease, whereas overexpression led to an increase in resistance of transgenic plants to Verticillium wilt. Interestingly, through binding to GhKWL1, the pathogenic effector protein VdISC1 produced by V. dahliae could impair the defense response mediated by GhKWL1. Therefore, our study suggests there is a GhKWL1-mediated defense response in cotton, which can be hijacked by V. dahliae through the interaction of VdISC1 with GhKWL1.     

Exploring the Effectiveness and Durability of Trans-Kingdom Silencing of Fungal Genes in the Vascular Pathogen Verticillium dahliae

Host-induced gene silencing (HIGS) based on trans-kingdom RNA interference (RNAi) has been successfully exploited to engineer host resistance to pests and pathogens, including fungi and oomycetes. However, revealing the mechanisms underlying trans-kingdom RNAi between hosts and pathogens lags behind applications. The effectiveness and durability of trans-kingdom silencing of pathogenic genes are uncharacterized. In this study, using our transgenic 35S-VdH1i cotton plants in which dsVdH1-derived small RNAs (siVdH1) accumulated, small RNA sequencing analysis revealed that siVdH1s exclusively occur within the double-stranded (ds)VdH1 region, and no transitive siRNAs were produced beyond this region in recovered hyphae of Verticillium dahliae (V. dahliae). Accordingly, we found that VdH1 silencing was reduced over time in recovered hyphae cultured in vitro, inferring that once the fungus got rid of the 35S-VdH1i cotton plants would gradually regain their pathogenicity. To explore whether continually exporting dsRNAs/siRNAs from transgenic plants into recipient fungal cells guaranteed the effectiveness and stability of HIGS, we created GFP/RFP double-labeled V. dahliae and transgenic Arabidopsis expressing dsGFP (35S-GFPi plants). Confocal images visually demonstrate the efficient silencing of GFP in V. dahliae that colonized host vascular tissues. Taken together, our results demonstrate that HIGS effectively triggers long-lasting trans-kingdom RNAi during plant vasculature V. dahliae interactions, despite no amplification or transitivity of RNAi being noted in this soil-borne fungal pathogen.     

MAT2A Localization and Its Independently Prognostic Relevance in Breast Cancer Patients

(1) Background: methionine cycle is not only essential for cancer cell proliferation but is also critical for metabolic reprogramming, a cancer hallmark. Hepatic and extrahepatic tissues methionine adenosyltransferases (MATs) are products of two genes, MAT1A and MAT2A that catalyze the formation of S-adenosylmethionine (SAM), the principal biological methyl donor. Glycine N-methyltransferase (GNMT) further utilizes SAM for sarcosine formation, thus it regulates the ratio of SAM:S-adenosylhomocysteine (SAH). (2) Methods: by analyzing the TCGA/GTEx datasets available within GEPIA2, we discovered that breast cancer patients with higher MAT2A had worse survival rate (p = 0.0057). Protein expression pattern of MAT1AA, MAT2A and GNMT were investigated in the tissue microarray in our own cohort (n = 252) by immunohistochemistry. MAT2A C/N expression ratio and cell invasion activity were further investigated in a panel of breast cancer cell lines. (3) Results: GNMT and MAT1A were detected in the cytoplasm, whereas MAT2A showed both cytoplasmic and nuclear immunoreactivity. Neither GNMT nor MAT1A protein expression was associated with patient survival rate in our cohort. Kaplan–Meier survival curves showed that a higher cytoplasmic/nuclear (C/N) MAT2A protein expression ratio correlated with poor overall survival (5 year survival rate: 93.7% vs. 83.3%, C/N ratio ≥ 1.0 vs. C/N ratio < 1.0, log-rank p = 0.004). Accordingly, a MAT2A C/N expression ratio ≥ 1.0 was determined as an independent risk factor by Cox regression analysis (hazard ratio = 2.771, p = 0.018, n = 252). In vitro studies found that breast cancer cell lines with a higher MAT2A C/N ratio were more invasive. (4) Conclusions: the subcellular localization of MAT2A may affect its functions, and elevated MAT2A C/N ratio in breast cancer cells is associated with increased invasiveness. MAT2A C/N expression ratio determined by IHC staining could serve as a novel independent prognostic marker for breast cancer.     

2—甲基—1—丁烯异构为2—甲基—2—丁烯的研究

异戊烯是抽余碳五的一种馏分，主要由两种同分异构体2—甲基—2—丁烯(2MB2)和2—甲基—1—丁烯(2MB1)组成，其中，2—甲基—2—丁烯含量越高，应用价值越高。本研究提供了一种提高异戊烯中2—甲基—2—丁烯含量的方法，即在催化剂作用下，在一定的温度、压力和空速下，以液相形式将2—甲基—1—丁烯异构为2—甲基—2—丁烯。最佳反应条件为：反应温度25—55℃，反应空速5—20hr^-1，反应压力0．6—0．9MPa。在此条件下，异戊烯中的2—甲基—2—丁烯与2—甲基—1—丁烯的比例由原料中的1—4：1提高到10—13：1。     

1-羟乙基-2-苯并咪唑氨基甲酸甲酯和1-乙基(2’一丙烯酸酯-1’-基)-2-苯并咪唑氨基甲酸甲酯的合成及用薄层一紫外分光光度法测定其含量

１－乙基（２’－丙烯酸酯－１’－基）－２－苯并咪唑氨基甲酸甲酯是合成多菌灵高 分子化学型缓释剂的重要单体。本文报道了１－乙基（２’－丙烯酸酯－１’－基）－２－苯并 咪唑氨基甲酸甲酯及其制备过程中的中间体－１－羟乙基－２－苯并咪唑氨基甲酸甲酯的 合成和分析方法。     

1-(2-氯苯基)-2-(4-氟苯基)-3-溴-1-丙烯的合成

1-(2-氯苯基)-2-(4-氟苯基)-3-溴-1-丙烯(2)是氟环唑的关键中间体。以1,2-二氯乙烷和水为溶剂,1-(2-氯苯基)-2-(4-氟苯基)-1-丙烯(1)在氢溴酸/双氧水体系中发生α-溴代反应得到该化合物。通过使用氢溴酸/双氧水代替溴素或N-溴代丁二酰亚胺,提高了溴的利用率及产品的收率,避免了烯烃的加成,减少了副反应的发生,减少了三废。通过实验获得了反应的优惠条件:二氯乙烷和水为溶剂,回流反应,n(1)∶n(HBr)∶n(H2O2)=1∶1.1∶2.5,氢溴酸和双氧水的滴加时间为6 h,在上述反应条件下,反应收率为90.6%,选择性90.6%。     

6-(呋喃-2-亚甲氨基)-2-甲基-1H-吲哚-3-羧酸乙酯的合成、表征与生物活性研究

以2,4-二硝基氯苯为原料,经过3步反应得到标题化合物。当温度为78℃,反应时间为8.5 h,目标化合物的收率可达65%。通过IR、1HNMR、13CNMR和MS光谱技术对其结构进行表征。运用SRB法评价了目标化合物对人体肺癌细胞的生长抑制活性。初步的生物实验表明,该化合物对癌细胞H1792、A549、H157和Calu-1具有生长抑制作用,且具有浓度依赖性。细胞周期分析发现其能诱导Calu-1细胞凋亡。     

2-(2-氯苄基)-2-(1-氯环丙基)环氧乙烷的合成

报道了丙硫菌唑中间体2-(2-氯苄基)-2-(1-氯环丙基)环氧乙烷的合成方法。以1-氯-N,N-二甲基环丙基甲酰胺和邻氯氯苄为原料,经格氏反应制备2-氯苄基-(1-氯环丙基)酮;后者再与硫叶立德试剂反应得到相应的环氧化物。     

2-甲基-3-(2-呋喃基)-2-丙烯-1-醇的合成研究

本文报道了利用交叉Cannizzaro反应,合成2-甲基-3-(2-呋喃基)-2-丙烯-1-醇的一种新方法。该合成法,步骤简单;原料易得,价廉;在最佳反应条件下,产率可达58．1％。较已有文献报道的产率44％有较大幅度的提高。     

羟磺甜菜碱的研制

研究了羟磺甜菜碱（ＣＤＳ）的合成工艺路线。研制产品与名牌进口样品的红外图谱显示完全一致,各项技术指标基本相符。     

1-(2-三甲基硅氧乙基)-5-巯基四氮唑的合成研究

以乙醇胺为起始原料,经三甲基氯硅烷保护制得2-(三甲基硅氧基)乙胺,再经与二硫化碳反应,再与溴乙烷缩合得到N-(2-三甲基硅氧乙基)二硫代氨基甲酸乙酯、然后再与叠氮化钠成环反应合成了1-(2-三甲基硅氧乙基)-5-巯基四氮唑.用三甲基氯硅烷作为乙醇胺的羟基保护试剂,保护和脱保护具有反应条件温和、收率高等 优点,反应总收...     

1-[2-(2,4-二氯苯基)-4-烃氧基甲基-1,3-二氧戊环-2-基]甲基-1H-1,2,4-三氮唑类化合物的合成及杀菌活性研究

2,4-二氯苯乙酮经溴化得ω-溴-2,4-二氯苯乙酮,该化合物与丙三醇在对甲基苯磺酸催化下脱水,得2-(2,4-二氯苯基)-2-溴甲基-4-羟甲基-1,3-二氧戊环,该中间体与苯甲酰氯在常温下反应,合成了2-(2,4-二氯苯基)-2-溴甲基-4-苯甲酰氧基甲基-1,3-二氧戊环,然后与三氮唑钠盐在130℃反应36 h,之后在碱性条件下水解,获得1-[2-(2,4-二氯苯基)-4-羟甲基-1,3-二氧戊环-2-基]甲基-1H-1,2,4-三氮唑,再以吡啶为缚酸剂继续与甲磺酰氯反应,合成了1-[2-(2,4-二氯苯基)-4-甲磺酰氧基甲基-1,3-二氧戊环-2-基]甲基-1H-1,2,4-三氮唑,最后在碱性条件下,与12个不同结构的酚缩合成标题化合物1-[2-(2,4-二氯苯基)-4-烃氧基甲基-1,3-二氧戊环-2-基]甲基-1H-1,2,4-三氮唑。标题化合物的结构用GC-MS、FTIR进行了表证。生物活性实验结果表明,12个标题化合物对水稻稻瘟病菌的抑菌率均在88.0%以上,其中,1-[2-(2,4-二氯苯基)-4-间甲基苯氧基甲基-1,3-二氧戊环-2-基]甲基-1H-1,2,4-三氮唑达100%。1-[2-(2,4-二氯苯基)-4-苯氧基甲基-1,3-二氧戊环-2-基]甲基-1H-1,2,4-三氮唑和1-[2-(2,4-二氯苯基)-4-对硝基苯氧基甲基-1,3-二氧戊环-2-基]甲基-1H-1,2,4-三氮唑对油菜菌核病菌的抑菌率分别为100%和97.8%;1-[2-(2,4-二氯苯基)-4-间甲基苯氧基甲基-1,3-二氧戊环-2-基]甲基-1H-1,2,4-三氮唑对小麦赤霉病菌的抑制活性为91.2%。     

1-[3-（三氟甲基）苯基]-2-丙酮合成的研究

以间-溴三氟甲苯为原料,经格氏反应和氧化反应合成1-[3-（三氟甲基）苯基]-2-丙酮。反应优化条件为：溴化亚铜为催化剂,反应时间3h,反应温度5℃,氧化时n（重铬酸钠）：n（浓硫酸）=1：4。在此优化条件下。总收率82．3％。产物结构经IR和^1H NMR确证。     

2,4,4-三甲基-3-(3-羰基-1-丁烯-1-基)-2-环己烯基-1-肟的合成研究

以环柠檬醛为原料,经过亚硝化、缩合两步反应得到2,4,4-三甲基-3-(3-羰基-1-丁烯-1-基)-2-环己烯基-1-肟,收率达65.1%。并分析了该反应中亚硝化的反应机理。     

2-己基-1,3-二噁戊烷的合成研究

在对甲苯磺酸和硫酸铜复合催化剂存在下,以正庚醛（Ａ）和乙二醇（Ｂ）为原料经脱水缩合合成了新型香料２－己基－１,３－二恶戊烷。优化试验结果表明：在ｎ（Ａ）：ｎ（Ｂ）＝１：１．１、催化剂用量为主原料总质量的０．２５％、反应时间为６ｈ坂应温度为９１～９８℃,产物收率达９４．１％。产物经理化检测和红外光谱确证。     

Downregulation of Methionine Cycle Genes MAT1A and GNMT Enriches Protein-Associated Translation Process and Worsens Hepatocellular Carcinoma Prognosis

The major biological methyl donor, S-adenosylmethionine (adoMet) synthesis occurs mainly in the liver. Methionine adenosyltransferase 1A (MAT1A) and glycine N-methyltransferase (GNMT) are two key enzymes involved in the functional implications of that variation. We collected 42 RNA-seq data from paired hepatocellular carcinoma (HCC) and its adjacent normal liver tissue from the Cancer Genome Atlas (TCGA). There was no mutation found in MAT1A or GNMT RNA in the 42 HCC patients. The 11,799 genes were annotated in the RNA-Seq data, and their expression levels were used to investigate the phenotypes of low MAT1A and low GNMT by Gene Set Enrichment Analysis (GSEA). The REACTOME_TRANSLATION gene set was enriched and visualized in a heatmap along with corresponding differences in gene expression between low MAT1A versus high MAT1A and low GNMT versus high GNMT. We identified 43 genes of the REACTOME_TRANSLATION gene set that are powerful prognosis factors in HCC. The significantly predicted genes were referred into eukaryotic translation initiation (EIF3B, EIF3K), eukaryotic translation elongation (EEF1D), and ribosomal proteins (RPs). Cell models expressing various MAT1A and GNMT proved that simultaneous restoring the expression of MAT1A and GNMT decreased cell proliferation, invasion, as well as the REACTOME_TRANSLATION gene EEF1D, consistent with a better prognosis in human HCC. We demonstrated new findings that downregulation or defect in MAT1A and GNMT genes can enrich the protein-associated translation process that may account for poor HCC prognosis. This is the first study demonstrated that MAT1A and GNMT, the 2 key enzymes involved in methionine cycle, could attenuate the function of ribosome translation. We propose a potential novel mechanism by which the diminished GNMT and MAT1A expression may confer poor prognosis for HCC.     

2-溴-2-硝基-1,3-二丙二醇的合成

以硝基甲烷和四醛为原料,在碱性条件下经烷基羟基化和溴化两步合成2-溴-2-硝基-1,3-二丙二醇,考虑了烷基羟基化反应,溴化反应温度,时间,对产品收率的影响.收率可达78.6%     

2-羟基-3-烯丙氧基丙磺酸钠的合成

采用新方法合成了2-羟基-3-烯丙氧基丙磺酸钠。先由环氧氯丙烷和亚硫酸氢钠合成中间体3-氯-2-羟基丙磺酸钠，然后中间体再与丙烯醇在氢氧化钠催化下生成目标产物2-羟基-3-烯丙氧基丙磺酸钠。2步反应总收率在70％左右，中间体和产物分别进行了光谱表征。     

溴化铜作溴化剂合成3-氯- α-溴-苯基乙基酮

以间氯苯基乙基酮和溴化铜为原料，用乙酸乙酯与氯仿组成的混合液作溶剂合成了3－氯－α－溴－苯基乙基酮。考察了影响溴代收率的因素，最佳合成条件：溴化铜：间氯苯基乙基酮（摩尔比）为2.1∶1，反应在回流温度下进行，反应时间2.5h，收率可达91.9％。