Heat Shock Factors HsfB 1 and HsfB2b Are Involved in the Regulation of Pdf1.2 Expression and Pathogen Resistance in Arabidopsis

In order to assess the functional roles of heat stress-induced class B-heat shock factors in Arabidopsis, we investigated T-DNA knockout mutants of AtHsfB1 and AtHsfB2b. Micorarray analysis of double knockout hsfB1/hsfB2b plants revealed as strong an up-regulation of the basal mRNA-levels of the defensin genes Pdfl.2a/b in mutant plants. The Pdfexpression was further enhanced by jasmonic acid treatment or infection with the necrotrophic fungus Alternaria brassicicola. The single mutant hsfB2b and the double mutant hsfB 1/B2b were significantly improved in disease resistance after A. brassicicola infection. There was no indication for a direct interaction of Hsf with the promoter of Pdf1.2, which is devoid of perfect HSE consensus Hsf-binding sequences. However, changes in the formation of late HsfA2-dependent HSE binding were detected in hsfB1/B2b plants. This suggests that HsfB1/B2b may interact with class A-Hsf in regulating the shut-off of the heat shock response. The identification of Pdfgenes as targets of Hsf-dependent negative regulation is the first evidence for an interconnection of Hsf in the regulation of biotic and abiotic responses.     

The Quaternary Structure of NADPH Thioredoxin Reductase C Is Redox-Sensitive

NADPH thioredoxin reductase C （NTRC） is a chloroplast enzyme able to conjugate NADPH thioredoxin reductase （NTR） and thioredoxin （TRX） activities for the efficient reduction of 2-Cys peroxiredoxin （2-Cys PRX）. Because NADPH can be produced in chloroplasts during darkness, NTRC plays a key role for plant peroxide detoxification during the night. Here, it is shown that the quaternary structure of NTRC is highly dependent on its redox status. In vitro, most of the enzyme adopted an oligomeric state that disaggregated in dimers upon addition of NADPH, NADH, or DTT. Gel filtration and Western blot analysis of protein extracts from Arabidopsis chloroplast stroma showed that native NTRC forms aggregates, which are sensitive to NADPH and DTT, suggesting that the aggregation state might be a significant aspect of NTRC activity in vivo. Moreover, the enzyme is localized in clusters in Arabidopsis chloroplasts. NTRC triple and double mutants, A164G- V182E-R183F and A164G-R183F, replacing key residues of NADPH binding site, showed reduced activity but were still able to dimerize though with an increase in intermediary forms. Based on these results, we propose that the catalytically active form of NTRC is the dimer, which formation is induced by NADPH.     

植物响应联合胁迫机制的研究进展

自然界中, 植物通常面对多重联合胁迫。在全球气候变化日益加剧的背景下, 多重联合胁迫对植物生长发育及作物产量形成的不利影响日益显著。阐明植物响应和适应联合胁迫的生理与分子机制, 对人们理解植物对自然环境的适应机理, 及培育耐受联合胁迫的新品种有重要意义。研究表明, 植物响应联合胁迫的机制是特异的, 不能简单地从单一胁迫响应叠加来推断。植物遭受联合胁迫时, 各种生理、代谢和信号途径相互作用, 使得植物响应联合胁迫非常复杂。该文综述了植物响应联合胁迫的生理与分子机理的最新进展, 并阐述了植物响应联合胁迫的研究方法。     

植物响应联合胁迫机制的研究进展

自然界中, 植物通常面对多重联合胁迫。在全球气候变化日益加剧的背景下, 多重联合胁迫对植物生长发育及作物产量形成的不利影响日益显著。阐明植物响应和适应联合胁迫的生理与分子机制, 对人们理解植物对自然环境的适应机理, 及培育耐受联合胁迫的新品种有重要意义。研究表明, 植物响应联合胁迫的机制是特异的, 不能简单地从单一胁迫响应叠加来推断。植物遭受联合胁迫时, 各种生理、代谢和信号途径相互作用, 使得植物响应联合胁迫非常复杂。该文综述了植物响应联合胁迫的生理与分子机理的最新进展, 并阐述了植物响应联合胁迫的研究方法。     

异源表达CkLEA1基因增强了拟南芥对非生物胁迫的耐受性

植物在生长过程中会受到各种非生物胁迫的伤害,导致生长发育和产量受到严重影响,胚胎晚期丰富蛋白（late embryogenesis abundant proteins,LEA蛋白）在植物抵抗非生物胁迫过程中起着重要的保护作用。在前期的研究基础上,将受多种胁迫诱导的柠条锦鸡儿CkLEA1（GenBank登录号KC309408）基因转入野生型拟南芥,通过实时荧光定量PCR从7株T₃代纯合体中筛选出3个转基因株系做进一步研究。种子萌发率实验发现,在200 mmol/L NaCl和400 mmol/L甘露醇处理下,转基因株系萌发率均高于野生型拟南芥。干旱处理2周大的幼苗后,转基因株系明显比野生型更抗旱,存活率高于野生型,并且失水率低于野生型。同时,转基因株系积累了较少的丙二醛（MDA）,超氧化物歧化酶（SOD）活性和谷胱甘肽（GSH）含量也高于野生型。这些结果表明,柠条锦鸡儿CkLEA1基因在种子萌发阶段提高了拟南芥对盐和渗透胁迫的耐受性,并且提高了转基因拟南芥幼苗生长阶段对干旱胁迫的抵抗能力。     

The N-terminal ATPase AT-hook-containing region of the Arabidopsis chromatin-remodeling protein SPLAYED is sufficient for biological activity

The SNF2-like chromatin-remodeling ATPase SPLAYED (SYD) was identified as a co-activator of floral homeotic gene expression in Arabidopsis. SYD is also required for meristem maintenance and regulates flowering under a non-inductive photoperiod. SNF2 ATPases are structurally and functionally conserved from yeast to humans. In addition to the conserved protein features, SYD has a large unique C-terminal domain. We show here that SYD is present as two forms in the nucleus, full-length and truncated, with the latter apparently lacking the C-terminal domain. The ratio of the two forms of endogenous SYD differs in juvenile and in adult tissues. Furthermore, an SYD variant lacking the C-terminal domain (SYDDeltaC) rescues the syd null mutant, indicating that the N-terminal ATPase AT-hook-containing region of SYD is sufficient for biological activity. Plants expressing SYDDeltaC show molecular and morphological phenotypes opposite to those of the null mutant, suggesting that the construct results in increased activity. This increased activity is at least in part due to elevated SYD protein levels in these lines. We propose that the C-terminal domain may control SYD accumulation and/or specific activity in the context of the full-length protein. The presence of the C-terminal domain in rice SYD suggests that its role is probably conserved in the two classes of flowering plants.     

干旱胁迫下沙生灌木花棒的抗氧化保护响应研究

采用盆栽试验,测定了在不同土壤水分胁迫程度和胁迫时间下花棒苗抗氧化保护酶(SOD、CAT、POD、APX)活性和抗氧化物(AsA、GSH、Car)含量,并用隶属函数值法综合评价了试验条件下花棒苗的抗氧化能力。结果显示:(1)与同期适宜水分处理相比,花棒苗的抗氧化保护酶活性在中度和重度干旱胁迫30d时均显著增加;胁迫60d时,POD活性均显著降低,而其余酶活性仍增加(重度胁迫)或显著增加(中度胁迫);胁迫90d时,中度胁迫处理除APX活性持平外,其余酶活性仍显著升高,而重度胁迫处理的SOD活性持平、POD活性显著增加、CAT和APX活性均显著降低。(2)相对于同期适宜水分处理,干旱胁迫30d时,中度胁迫处理的花棒苗AsA、Car、GSH含量分别显著增加、稍有增加和显著降低,而重度胁迫的AsA显著增加,Car和GSH含量均显著降低;胁迫60d时,中度胁迫处理的AsA、Car、GSH含量分别显著增加、稍有降低和稍有提高,而重度胁迫的AsA、Car、GSH含量分别显著增加、稍有增加和显著降低;胁迫90d时,中度胁迫处理的AsA含量显著增加,Car和GSH含量均稍有提高,而重度胁迫的AsA含量稍有下降,Car和GSH含量则持平。(3)花棒苗的MDA含量在干旱胁迫过程中均比适宜水分处理增加,且中后期达到显著水平。(4)花棒苗抗氧化能力在干旱胁迫30d表现为重度干旱>中度干旱>适宜水分,胁迫60d为中度干旱>重度干旱>适宜水分,胁迫90d时为中度干旱>适宜水分>重度干旱。研究表明,花棒苗在中度干旱胁迫下能调动自身抗氧化保护酶类与抗氧化物质并提高其抗氧化能力,消除或减轻干旱诱导的活性氧毒害;重度干旱胁迫下的花棒苗在胁迫前中期亦能增强自身的抗氧化能力,但在胁迫后期因部分活性氧清除机制受到破坏而使抗氧化能力降低。     

大叶杨干旱和水淹胁迫的转录组分析

大叶杨（Populus lasiocarpa）是中国特有的杨属物种,干旱和水淹是影响大叶杨生长和分布范围的两个关键因子。AP2/ERF转录因子家族在植物响应非生物胁迫中发挥重要作用。本研究采用转录组测序、生物信息学分析手段并结合分子实验验证初步鉴定了参与大叶杨干旱和水淹胁迫响应的关键基因。研究结果显示：（1）在大叶杨中分别鉴定到3,986/385个响应干旱/水淹胁迫的差异表达基因,其中包括237个同时响应干旱和水淹胁迫的差异表达基因。（2）在大叶杨中共鉴定到205个AP2/ERF家族成员,系统发育分析表明其在大叶杨中主要分为5个亚家族,并显著富集于差异表达基因中。（3）筛选部分胁迫前后差异表达的PlAP2/ERF基因进行qRT-PCR实验,经证实这些基因在大叶杨受到干旱/水淹胁迫时均可被诱导表达。综上,大叶杨在水淹胁迫下的差异表达基因数量明显少于干旱胁迫,AP2/ERF基因家族的部分基因参与到大叶杨干旱/水淹胁迫的应激表达过程。     

表观遗传调控植物响应非生物胁迫的研究进展

植物的生长发育容易受到外界环境变化的影响。非生物胁迫发生时, 表观遗传机制对胁迫应答基因的表达调控发挥了十分重要的作用。近年来, 调控植物非生物胁迫应答的表观遗传机制研究取得了一系列重要进展, 为进一步深入解析植物响应非生物胁迫的分子机制奠定了基础。该文对DNA甲基化修饰、组蛋白修饰、染色质重塑和非编码RNA等主要表观遗传调控方式在植物响应非生物胁迫中的作用进行了简要综述。     

Defining the Functional Network of Epigenetic Regulators in Arabidopsis thaliana

Development of ChiP-chip and ChlP-seq technologies has allowed genome-wide high-resolution profiling of chromatin-associated marks and binding sites for epigenetic regulators. However, signals for directing epigenetic modifiers to their target sites are not understood. In this paper, we tested the hypothesis that genome location can affect the involvement of epigenetic regulators using Chromatin Charting （CC） Lines, which have an identical transgene construct inserted at different locations in the Arabidopsis genome. Four CC lines that showed evidence for epigenetic silencing of the luciferase reporter gene were transformed with RNAi vectors individually targeting epigenetic regulators LHP1, MOM1, CMT3, DRD1, DRM2, SUVH2, CLF, and HD1. Involvement of a particular epigenetic regulator in silencing the transgene locus in a CC line was determined by significant alterations in luciferase expression after suppression of the regulator＇s expression. Our results suggest that the targeting of epigenetic regulators can be influenced by genome location as well as sequence context. In addition, the relative importance of an epigenetic regulator can be influenced by tissue identity. We also report a novel approach to predict interactions between epigenetic regulators through clustering analysis of the regulators using alterations in gene expression of putative downstream targets, including endogenous loci and transgenes, in epigenetic mutants or RNAi lines. Our data support the existence of a complex and dynamic network of epigenetic regulators that serves to coordinate and control global gene expression in higher plants.     

4种沙生灌木幼苗PV曲线水分参数对干旱胁迫的响应

以柠条、沙木蓼、杨柴和花棒4种沙生灌木幼苗为材料,采用盆栽法在适宜水分、中度干旱和重度干旱(田间持水量的75%、50%和35%)3种土壤水分条件下,应用PV技术测定了它们在膨压为0时的渗透势(ψstlp)、相对水含量(RWCtlp)和相对渗透水含量(ROWCtlp),以及饱和含水时的渗透势(ψssat)、束缚水含量(Va)、膨压随叶水势下降而降低的速率b值和组织细胞总体弹性模量(ε′)等水分参数,并用隶属函数值法对4种苗木在干旱下保持膨压的能力进行了综合评价.结果显示:与适宜水分条件下相比较,除花棒幼苗在中度及重度干旱下ψssat值、柠条苗在中度干旱下ψssat和ψstlp差值和在中度及重度干旱下Va值、沙木蓼苗在中度干旱下ε′和RWCtlp值均变化很小以外,4种苗木其它水分参数在不同程度干旱胁迫下均有较明显的变化,且其变化幅度随干旱胁迫的加剧而增加,从而使苗木保持膨压及吸水保水的能力较适宜水分下明显增强;苗木保持膨压能力的综合评价结果为:中度干旱下柠条>花棒>杨柴>沙木蓼,重度干旱下柠条>花棒>沙木蓼>杨柴.研究表明,柠条幼苗对干旱胁迫具有极强的渗透调节适应性.     

植物气孔对全球环境变化的响应及其调控防御机制

气孔是植物与环境发生联系的重要门户,控制着植物与外界的气体和水分交换。本文针对全球大气CO2浓度升高、气候变暖、干旱加剧等环境问题,分析了气孔对全球水循环、碳循环的重要贡献。系统总结了气孔的形态发育和生理功能对大气高CO2浓度、干旱、土壤盐渍化、病虫害等的响应及其调控防御机制。综述了脱落酸（ABA）、Ca2＋、H2O2、一氧化氮（NO）和光信号调控气孔运动的分子机制。从理论和实践两方面,提出了通过调控气孔运动协调CO2同化和水分散失的矛盾,在不影响光合效率的前提下提高水分利用率等未来的研究方向。     

拟南芥AtR8 lncRNA对盐胁迫响应及其对种子萌发的调节作用

长链非编码RNA (lncRNA)是一类长度大于200个核苷酸且不编码蛋白质的非编码RNA, 主要由RNA聚合酶II转录生成, 大量存在于生物体内并具有多种生物学功能。AtR8 lncRNA是拟南芥(Arabidopsis thaliana)中RNA聚合酶III转录的长链非编码RNA。前期研究发现, 水杨酸(SA)处理诱导萌发种子中AtR8 lncRNA的表达, AtR8 lncRNA缺失抑制SA胁迫下的种子萌发。进一步研究发现, AtR8 lncRNA转录区域内存在保守的盐胁迫响应元件(TCTTCTTCTTTA); NaCl处理抑制萌发种子中AtR8 lncRNA的表达; 与野生型相比, 高浓度NaCl处理明显抑制了atr8 (AtR8 lncRNA部分缺失型拟南芥)种子萌发。研究结果表明, AtR8 lncRNA在拟南芥种子萌发期的盐胁迫中起重要作用。