Direct ubiquitination of pattern recognition receptor FLS2 attenuates plant innate immunity

Innate immune responses are triggered by the activation of pattern-recognition receptors (PRRs). The Arabidopsis PRR FLAGELLIN-SENSING 2 (FLS2) senses bacterial flagellin and initiates immune signaling through association with BAK1. The molecular mechanisms underlying the attenuation of FLS2 activation are largely unknown. We report that flagellin induces recruitment of two closely related U-box E3 ubiquitin ligases, PUB12 and PUB13, to FLS2 receptor complex in Arabidopsis. BAK1 phosphorylates PUB12 and PUB13 and is required for FLS2-PUB12/13 association. PUB12 and PUB13 polyubiquitinate FLS2 and promote flagellin-induced FLS2 degradation, and the pub12 and pub13 mutants displayed elevated immune responses to flagellin treatment. Our study has revealed a unique regulatory circuit of direct ubiquitination and turnover of FLS2 by BAK1-mediated phosphorylation and recruitment of specific E3 ligases for attenuation of immune signaling.     

Concentration difference of auxin involved in stem development in soybean

Auxin regulates cell division and elongation of the primordial cells through its concentration and then shaped the plant architecture. Cell division and elongation form the internode of soybean and result in different plant heights and lodging resistance. Yet the mechanisms behind are unclear in soybean. To elucidate the mechanism of the concentration difference of auxin related to stem development in soybean, samples of apical shoot, elongation zone, and mature zone from the developing stems of soybean seedlings, Charleston, were harvested and measured for auxin concentration distributions and metabolites to identify the common underlying mechanisms responsible for concentration difference of auxin. Distribution of indole~(-3)-acetic acid(IAA), indole~(-3)-butyric acid(IBA), and methylindole~(-3)-acetic acid(Me-IAA) were determined and auxin concentration distributions were found to have a complex regulation mechanism. The concentrations of IAA and Me-IAA in apical shoot were significantly different between elongation zone and mature zone resulting in an IAA gradient. Tryptophan dependent pathway from tryptamine directly to IAA or through indole~(-3)-acetonitrile to IAA and from indole~(-3)-propionic acid(IPA) to IAA were three primary IAA synthesis pathways. Moreover, some plant metabolites from flavonoid and phenylpropanoid synthesis pathways showed similar or reverse gradient and should involve in auxin homeostasis and concentration difference. All the data give the first insight in the concentration difference and homeostasis of auxin in soybean seedlings and facilitate a deeper understanding of the molecular mechanism of stem development and growth. The gathered information also helps to elucidate how plant height is formed in soybean and what strategy should be adopted to regulate the lodging resistance in soybean.     

猪圆环病毒1型ORF2基因的克隆与序列分析

【研究目的】提取沙门氏菌鞭毛蛋白并鉴定其抗原性;【方法】选用鸭沙门氏菌经半固体培养基诱导鞭毛,经肉汤培养后,用酸裂解、经差速离心、硫酸铵沉淀,分离出粗制鞭毛蛋白。将粗制的鞭毛蛋白用SDS-PAGE和磷钨酸负染投射电镜观察,并包板做ELISA检测其抗原性;【结果】SDS-PAGE显示提取的鞭毛蛋白相对分子质量约为58.0KD,且纯度较高,抗原性较好,每1000ml培养液可获得32.8mg鞭毛蛋白;【结论】酸裂解法可获得高质量的鞭毛蛋白,抗原性较好,为禽副伤寒沙门氏菌病诊断方法的建立奠定了基础。     

PIN proteins perform a rate-limiting function in cellular auxin efflux

Intercellular flow of the phytohormone auxin underpins multiple developmental processes in plants. Plant-specific pin-formed (PIN) proteins and several phosphoglycoprotein (PGP) transporters are crucial factors in auxin transport-related development, yet the molecular function of PINs remains unknown. Here, we show that PINs mediate auxin efflux from mammalian and yeast cells without needing additional plant-specific factors. Conditional gain-of-function alleles and quantitative measurements of auxin accumulation in Arabidopsis and tobacco cultured cells revealed that the action of PINs in auxin efflux is distinct from PGP, rate-limiting, specific to auxins, and sensitive to auxin transport inhibitors. This suggests a direct involvement of PINs in catalyzing cellular auxin efflux.     

超量表达CsGH3.6通过抑制生长素信号转导 增强柑橘溃疡病抗性

背景 由柑橘黄单胞杆菌柑橘亚种（Xanthomonas citri subsp. citri,Xcc）引起的柑橘溃疡病是柑橘生产上最具毁灭性的一种病害。植物生长素在调控柑橘溃疡病菌引起的寄主侵染部位脓疱形成中起重要作用。生长素早期响应基因GH3通过酰基化吲哚-3-乙酸（indole-3-acetic acid,IAA）调控植物激素动态平衡。前期研究发现柑橘CsGH3.6在调控生长素响应溃疡病侵染中起着重要作用。目的 通过对超量表达CsGH3.6转基因晚锦橙的抗病性、植株表型、细胞和激素变化进行分析,利用RNA-Seq解析CsGH3.6调控的信号通路,探明CsGH3.6调控激素动态平衡影响柑橘溃疡病抗性的内在机制。方法 利用针刺法对离体转基因叶片接种溃疡病菌Xcc,统计接种第10 天时病斑面积和病情指数,以野生型为对照,评价转基因植株的抗性水平;提取感病前后叶片内源激素,利用高效液相色谱技术(high performance liquid chromatography,HPLC )检测转基因植株中激素含量变化;温室中观察转基因植株表型变化;通过测量叶片纵径、横径和厚度分析转基因植株叶型变化特征;制备叶片表皮切片,显微观察表皮细胞和气孔,并统计转基因植株表皮细胞长度和气孔密度;采用RNA-Seq测序技术研究转基因植株转录组变化情况,并利用Nr、Nt、Pfam、COG、SwissProt和gene ontology (GO)数据库注释基因功能,进一步利用KEEG数据库和MapMan软件解析超量表达CsGH3.6影响的重要基因、功能和途径,阐明CsGH3.6调控柑橘溃疡病抗性的分子机制。结果 超量表达CsGH3.6显著增强转基因植株的溃疡病抗性;转基因植株分枝增多且下垂,叶片向上卷曲,变小,颜色浅;转基因植株气孔密度增加,表皮细胞变短;激素含量分析显示,转基因植株自由生长素（IAA）和茉莉酸（jasmonic acid,JA）含量显著降低,而水杨酸（salicylic acid,SA）含量显著增加;转录组测序分析表明,超量表达CsGH3.6显著抑制生长素信号转导相关基因表达,特别是所有注释的Aux/IAA家族基因均下调表达,相反,与生物胁迫相关基因的表达为上调,其中绝大部分基因为病程相关蛋白基因。结论 超量表达CsGH3.6通过酰基化自由IAA抑制生长素信号转导,调控JA和SA的动态平衡,改变细胞和植株的形态建成,从而增强柑橘对溃疡病的抗性。研究结果暗示调控激素动态平衡在柑橘抗病育种中具有潜在价值。     

The HD-Zip transcription factor GhHB12 represses plant height by regulating the auxin signaling in cotton

Upland cotton (Gossypium hirsutum L.) is the most important natural textile fiber crop worldwide. Plant height (PH) is a significant component of plant architecture, strongly influencing crop cultivation patterns, overall yield, and economic coefficient. However, cotton genes regulating plant height have not been fully identified. Previously, an HD-Zip gene (GhHB12) was isolated and characterized in cotton, which regulates the abiotic and biotic stress responses and the growth and development processes. In this study, we showed that GhHB12 was induced by auxin. Moreover, overexpression of GhHB12 induces the expression of HY5, ATH1, and HAT4, represses the spatial-temporal distribution, polar transport, and signaling of auxin, alters the expression of genes involved in cell wall expansion, and restrains the plant height in cotton. These results suggest a role of GhHB12 in regulating cotton plant height, which could be achieved by affecting the auxin signaling and cell wall expansion.     

SLPIN6基因沉默对番茄抗旱性的影响

SLPIN6基因是番茄PIN家族一员,番茄PIN家族是一类生长素运输体蛋白家族,调控生长素极性运输.研究表明,干旱胁迫改变植物生长素极性运输平衡,影响植物抗旱性.团队前期研究发现SLPIN6基因在干旱胁迫时表达量显著上调,可能参与番茄植株抗旱应答调控.为进一步明确该基因抗旱调节功能,试验采用VIGS方法对SLPIN6基因作沉默处理,通过观察番茄植株抗旱表型及测定抗逆生理指标变化,发现SLPIN6基因沉默后番茄植株与对照相比,沉默番茄植株萎蔫程度加重,抗逆生理指标差异明显,表明SLPIN6基因参与番茄抗旱应答调控,具有抗旱调节功能.     

Reversal of obesity- and diet-induced insulin resistance with salicylates or targeted disruption of Ikkbeta

We show that high doses of salicylates reverse hyperglycemia, hyperinsulinemia, and dyslipidemia in obese rodents by sensitizing insulin signaling. Activation or overexpression of the IkappaB kinase beta (IKKbeta) attenuated insulin signaling in cultured cells, whereas IKKbeta inhibition reversed insulin resistance. Thus, IKKbeta, rather than the cyclooxygenases, appears to be the relevant molecular target. Heterozygous deletion (Ikkbeta+/-) protected against the development of insulin resistance during high-fat feeding and in obese Lep(ob/ob) mice. These findings implicate an inflammatory process in the pathogenesis of insulin resistance in obesity and type 2 diabetes mellitus and identify the IKKbeta pathway as a target for insulin sensitization.     

金叶榆二倍体及其同源四倍体的生理特征与转录组差异分析

本文旨在探究金叶榆（Ulmus pumila cv.‘Jinye’）二倍体及其同源四倍体的生理特征和差异基因表达情况,为金叶榆关键调控基因的定位及基因工程育种提供分子基础。试验以金叶榆二倍体及其同源四倍体为材料,比较两者叶绿素相对含量,可溶性糖、可溶性蛋白、MDA含量,SOD和POD活性等生理指标,并进行转录组测序。结果表明：金叶榆同源四倍体叶片中叶绿素相对含量,可溶性糖和可溶性蛋白含量及SOD、POD活性显著高于二倍体,MDA含量显著低于二倍体。转录组测序共得到1 283个差异表达基因,其中1 078个表达上调,205个表达下调。995个差异表达基因被GO功能注释,与代谢过程、细胞过程、催化活性、结合和转运蛋白活性等密切相关;COG注释结果显示,主要注释在碳水化合物的运输和代谢、次生代谢产物合成和脂质运输和代谢;KEGG通路分析发现,植物激素信号转导通路差异显著性最为明显,其中生长素、油菜素内酯和水杨酸信号转导途径富集基因数最多。参与代谢过程、催化活性和激素信号转导通路等关键基因的差异表达,可能与同源四倍体植株能源物质含量高、酶活提高及生长势和抗逆性增强有关。     

黄曲霉毒素B1诱导MEK/ERK/ROS介导的巨噬细胞自噬反应

为研究黄曲霉毒素B1(AFB1)能否诱导细胞自噬,并初步探讨其可能的作用机制。利用免疫荧光、酶标仪检测、免疫印迹等技术检测经AFB1处理过的细胞的自噬水平、相关蛋白的表达量以及活性氧(Reactive oxygen species,ROS)的产生情况。结果表明:AFB1能够引起RAW264.7细胞和Hela细胞的自噬反应,并且该自噬为完全自噬。AFB1诱导的小鼠单核巨噬细胞RAW264.7自噬是依赖细胞外信号调节激酶(Extracellular signal-regulated kinase,ERK)途径发生的,并受ROS的调节。此外,发现AFB1诱导ROS产生呈浓度依赖性,而ROS抑制剂同时抑制了ROS和自噬发生,但自噬抑制剂并没有有效地抑制ROS的产生,表明ROS处于AFB1诱导自噬的上游,对自噬起到调节作用。这表明AFB1诱导的自噬现象在细胞中普遍存在,而自噬作为机体的免疫机制,在抵抗AFB1毒力侵染的过程中发挥重要作用。     

IAA分解代谢相关酶（IAAO、POD）的研究进展

 吲哚乙酸IAA参与植物体内诸多生理活动,其分解代谢相关酶吲哚乙酸氧化酶（IAAO）、过氧化物酶（POD）在IAA代谢过程中起着关键作用,二者通过调控植物体内IAA的水平来调控复杂的植物生长,该文综合总结了IAAO和POD近年来研究的相关进展,尤其是结合生物化学和分子生物学方面阐述了这两种酶在IAA代谢中的作用及其相关性。     

大豆疫霉效应因子PsCRN115诱导烟草抗性的机制分析

CRN(crinkling and necrosis induced protein)蛋白是卵菌所特有的一类效应因子,在已经测序的疫霉种中,每个基因组中预测都含有上百个CRN基因,关于其功能和分子机制目前研究不多。前期研究发现,大豆疫霉效应因子PsCRN115是大豆疫霉的致病性必需因子且能抑制植物的细胞死亡。为了进一步研究PsCRN115的功能,本研究采用农杆菌渗入法介导的基因瞬时转化技术在烟草上过表达该基因,发现PsCRN115可显著提高烟草对烟草疫霉的抗性。随后采用荧光定量PCR研究了其作用机制,发现PsCRN115可诱导水杨酸(SA)通路防卫反应基因的高表达,但对乙烯(ET)和茉莉酸(JA)通路防卫反应基因的表达影响不显著。表明:大豆疫霉效应因子PsCRN115是致病必需的因子,能被植物所识别,并诱导植物的防卫反应。     

NBS-LRR Proteins and Their Partners:Molecular Switches of Plant Defense

Specificity of the plant innate immune system is often conferred by resistance（R）proteins.Most plant disease resistance （R）proteins contain a series of leucine-rich repeats（LRRs）,a nucleotide-binding site（NBS）,and a putative amino-terminal signaling domain.They are termed NBS-LRR proteins.The LRRs are mainly involved in recognition,and the amino-terminal domain determines signaling specificity,whereas the NBS domain presumably functions as a molecular switch.During the past years,the most important discoveries are the role of partners in NBS-LRR gene mediated defenses,mounting support for the so-called＂guard hypothesis＂of R gene function,and providing evidence for intramolecular interactions and intermolecular interactions within NBS- LRR proteins as a mode of signaling regulation.The outcome of these interactions determines whether a plant activates its defense responses.     

An agonist of toll-like receptor 5 has radioprotective activity in mouse and primate models

The toxicity of ionizing radiation is associated with massive apoptosis in radiosensitive organs. Here, we investigate whether a drug that activates a signaling mechanism used by tumor cells to suppress apoptosis can protect healthy cells from the harmful effects of radiation. We studied CBLB502, a polypeptide drug derived from Salmonella flagellin that binds to Toll-like receptor 5 (TLR5) and activates nuclear factor-kappaB signaling. A single injection of CBLB502 before lethal total-body irradiation protected mice from both gastrointestinal and hematopoietic acute radiation syndromes and resulted in improved survival. CBLB502 injected after irradiation also enhanced survival, but at lower radiation doses. It is noteworthy that the drug did not decrease tumor radiosensitivity in mouse models. CBLB502 also showed radioprotective activity in lethally irradiated rhesus monkeys. Thus, TLR5 agonists could potentially improve the therapeutic index of cancer radiotherapy and serve as biological protectants in radiation emergencies.     

Role of the Arabidopsis glucose sensor HXK1 in nutrient,light, and hormonal signaling

Glucose modulates many vital processes in photosynthetic plants. Analyses of Arabidopsis glucose insensitive2 (gin2) mutants define the physiological functions of a specific hexokinase (HXK1) in the plant glucose-signaling network. HXK1 coordinates intrinsic signals with extrinsic light intensity. HXK1 mutants lacking catalytic activity still support various signaling functions in gene expression, cell proliferation, root and inflorescence growth, and leaf expansion and senescence, thus demonstrating the uncoupling of glucose signaling from glucose metabolism. The gin2 mutants are also insensitive to auxin and hypersensitive to cytokinin. Plants use HXK as a glucose sensor to interrelate nutrient, light, and hormone signaling networks for controlling growth and development in response to the changing environment.     

植物矮化基因相关研究进展

植物株型的矮化调控是遗传育种的一项热门研究。国内外学者对植物矮化机理、矮化基因、矮化遗传育种等均进行了较深入的研究,水稻、玉米、小麦等粮食作物和黄瓜、番茄、南瓜等园艺作物均已形成了较完善的矮化研究体系。矮化植株株型紧凑、冠幅小,能够有效提高抗倒伏能力,在生产实践中具有管理便利的优点,因此矮化育种是植物育种的发展趋势。激素调控是目前运用较为广泛的矮化调控手段,植物激素通过影响细胞的分裂和伸长来改变节间长度和数目,从而调节高度,达到矮化植株的效果,常用激素有赤霉素、油菜素内酯、生长素、乙烯等,这些激素促进或抑制植物的生长发育,与矮化突变体的形成有关,且各种激素信号通路之间存在相互作用。综述了禾本科、茄科、葫芦科等植物矮化基因的研究现状,激素调控下矮化突变体的形成,矮化基因的克隆及功能研究进展,探讨了植物矮生性状分子机理和分子遗传学研究进展,为后续研究植物矮化基因提供理论基础。     

生长素促进拟南芥AtNRT1.1基因表达增强硝酸盐吸收

为研究生长素信号对植物硝酸盐营养吸收的调控作用,采用扫描离子选择电极技术 (SIET) 和实时荧光定量 PCR 技术(qRT-PCR),测定拟南芥野生型Col-0、生长素过表达突变体yuc1-D以及生长素通路缺失突变体axr1-12 3种株系初始根中硝酸根离子(NO-3)吸收速率以及硝酸盐转运蛋白基因AtNRT1.1表达量的差异,并进一步检测Col-0株系以及硝酸盐转运蛋白突变体nrt1.1株系在正常条件(CK)或施加外源IAA及生长素极性运输抑制剂2,3,5 三碘苯甲酸(TIBA)处理下的NO-3流速和AtNRT1.1基因表达量的差异。结果表明:yuc1 D株系的NO-3吸收速率以及AtNRT1.1基因表达量相比野生型均有大幅增加,而axr1-12株系的 NO-3吸收速率以及AtNRT1.1基因表达量相比野生型显著降低;在Col-0株系中施加外源IAA对NO-3吸收速率以及AtNRT1.1基因表达量有明显促进作用,而施加TIBA的效果反之,说明生长素对硝酸盐吸收有增强效应。nrt1.1株系在CK、IAA、TIBA处理下NO-3吸收速率差异较野生型不明显,揭示了AtNRT1.1基因在生长素促进硝酸盐吸收途径中所起的重要作用。      

Functional polymorphism among members of abscisic acid receptor family(ZmPYL) in maize

Pyrabactin resistance 1-like proteins(PYLs) are direct receptors of abscisic acid(ABA). For the redundant and polymorphic functions, some members of the PYL family interact with components of other signaling pathways. Here, 253 positive colonies from a maize cDNA library were screened as interacting proteins with the members of ZmPYL family. After sequencing and function annotation, 17 of 28 interaction combinations were verified by yeast two-hybrid(Y2 H). The germination potential, taproot length and proline content of a quartet mutant of Arabidopsis PYL genes were significantly deceased comparing to the wild type(WT) under alkaline stress(pH 8.5) and 100 μmol L–1 methyl jasmonate(MeJA) induction. The malondialdehyde(MDA) content was significantly increased. After germinating in darkness, the characteristics of dark morphogenesis of the quartet mutant seedlings were more obvious than those of the WT. The differential expression of the related genes of photomorphogenesis in the mutant was much more than that in the WT. Three light and two JA responsive cis-affecting elements were identified during the promoter sequences of the AtPYL1 and AtPYL2 genes. These results suggested that functional polymorphism has evolved among the members of ZmPYL family. In response to developmental and environmental stimuli, they not only function as direct ABA receptors but also interact with components of other signaling pathways mediated JA, brassinosteroid(BR), auxin, etc., and even directly regulate downstream stress-related proteins. These signaling pathways can interact at various crosstalk points and different levels of gene expression within a sophisticated network.