Proteomic analysis of pathogen-responsive proteins from maize stem apoplast triggered by Fusarium verticillioides

During the attack of a pathogen, a variety of defense-associated proteins are released by the host plant in the apoplast to impede the perceived attack. This study utilized the mass spectrometry(LC-MS/MS) and label-free quantification method to analyze the apoplastic fluid(APF) from maize stalk and identified the proteins responsive to the Fusarium verticillioides infection. We have identified 742 proteins, and among these, 119 proteins were differentially accumulated(DAPs), i.e., 35 up-regulated, 18 down-regulated, and 66 proteins were only induced by the pathogen infection. The differentially accumulated proteins were analyzed for their Gene Ontology(GO) and Kyoto Encyclopedia of Gene and Genomes(KEGG) pathway enrichment. The highly enriched Biological Process(BP) term was the L-serine biosynthesis process, whereas the most enriched Molecular Function(MF) term was the cysteine-type endopeptidase inhibitor activity. It was also found that the pathways related to the biosynthesis of amino acid, biosynthesis of secondary metabolites, protein processing in the endoplasmic reticulum, and carbohydrate metabolic pathways were significantly enriched. Moreover, 61 out of 119 differentially accumulated proteins were predicted as secretory proteins. The secretory pathways analysis showed that a greater number of proteins were secreted through the conventional secretion system compared to the unconventional secretion system. The identified secreted proteins were related to a variety of pathways in defense responses including cell redox homeostasis, cell wall modification, signal transduction, carbohydrate metabolism, binding proteins(metal ion binding, RNA binding and heme-binding), maintenance and stabilization of other proteins, indicating a complex response from the plant to the fungal infection. Our data suggested that a number of host proteins belonging to various pathways have been modulated in the apoplastic region.

Accumulation of Sugars and Liquid in Apoplast of Fruit Flesh Result in Pineapple Translucency

Translucency is a recurring problem for pineapple industry. Translucent fruit contained more sucrose, glucose and fructose in apoplast than those in apoplast of normal fruit. There were more liquid in intercellular space of translucent fruit than that of normal flesh. The contents of alcohol and ethylene in translucent fruit were higher than those in normal fruit. Translucent fruit contained less calcium than normal fruit. Electrolyte leakage of translucent flesh was more than that of normal flesh. There were 205 proteins of which the expressions in translucent flesh were higher than those in normal flesh. Calcium-ions-binding protein EF-hand domain-containing protein, ethylene-synthesizing enzyme 1-aminpcyclopropane-1-carboxylate oxidase, ROS-producing protein universal stress protein A-like protein were the top three proteins of which the expressions in translucent flesh were higher than those in normal fruit. When much sugar was transferred into fruit pulp and accumulated in intercellular space, water will be absorbed from cells around and translucence formed. The accumulation of sugar and liquid in apoplast were due to that cell wall and membrane were degraded, which was from being attacked by ROS. There might be more and larger pores in cell wall and membranes of translucent flesh. These data played foundations for researching methods for controlling pineapple translucency.

The pH of the Apoplast： Dynamic Factor with =unctional Impact Under Stress

apoplast 是有一部薄水电影的互连的分隔空间在应力下面的那 alkalinizes。这 ? 没有功能的含意，全身的 pH 增加可以是第二等的效果，从离子运动或质子泵规定产生。在另一方面，正在增加承受应力是机制的部分的指示。不管这争吵， apoplast 的 alkalinization 几乎没收到很少注意。apoplastic pH (pH apo ) 不仅在植物病原体相互作用期间而且响应咸度或干旱增加。不很对机制被知道引起叶 apoplast 到 alkalinize，也不是否，并且如果那么，功能的影响怎么被传送。争论解释被给了，并且 pH apo 规定在知识的这缺乏后面被看作主要原因。一聚在一起散布信息表明那在 pH apo 由经由在 abscisic 酸上施加的效果调整有气孔的孔传送功能。而且， apoplastic alkalinization 可以在应力下面调整生长，而这需要被验证。在这评论，全面调查在应力下面的 apoplast 被给的使成碱性，和 apoplastic alkalinization 的适用性作为为感觉信息的传播的 transducing 元素被讨论的大约几生理的机制。

Iron fractions in the apoplast of intact root tips of Zea mays L. seedlings affected by nitrogen form

The effects of ammonium (NH4+-N) and nitrate (NO3- - N) were examined on Fe fractions and FeCN (ferricyanide) reductase activity in intact root tips (0-3 cm) of young maize (Zea mays L. cv. Lenz) in solution culture by using short-term experiment under controlled Fe deficiency conditions (containing high HCO3- concentration in pre-culture solution). The results showed that Fe(Ⅱ) concentrations in root tip apoplast of maize were only 20-40 nmol/g FW which accounted for 7%-13% of total Fe. Most of Fe in root tips existed as Fe(Ⅲ) compounds. Imposition of the roots to NH4+ - N or NO3- - N for 60 min led to an increase of Fe(Ⅱ) in root tip apoplast. NH4+ - N led to an increased concentration of Fe(Ⅱ) and exchangeable Fe (Fe(Ⅱ) and Fe (Ⅲ)) in root tips, while NO3- - N increased FeCN reductase activity. The relationship between pH and Fe fractions, FeCN reductase activity was also discussed.

Apoplastic Proteases: Powerful Weapons against Pathogen Infection in Plants

Plants associate with diverse microbes that exert beneficial,neutral,or pathogenic effects inside the host.During the initial stages of invasion,the plant apoplast constitutes a hospitable environment for invading microbes,providing both water and nutrients.In response to microbial infection,a number of secreted proteins from host cells accumulate in the apoplastic space,which is related to microbial association or colonization processes.However,the molecular mechanisms underlying plant modulation of the apoplast environment and how plant-secreted proteases are involved in pathogen resistance are still poorly understood.Recently,several studies have reported the roles of apoplastic proteases in plant resistance against bacteria,fungi,and oomycetes.On the other hand,microbe-secreted proteins directly and/or indirectly inhibit host-derived apoplastic proteases to promote infection.These findings illustrate the importance of apoplastic proteases in plant–microbe interactions.Therefore,understanding the protease-mediated apoplastic battle between hosts and pathogens is of fundamental importance for understanding plant–pathogen interactions.Here,we provide an overviewof plant–microbe interactions in the apoplastic space.We define the apoplast,summarize the physical and chemical properties of these structures,and discuss the roles of plant apoplastic proteases and pathogen protease inhibitors in host–microbe interactions.Challenges and future perspectives for research into protease-mediated apoplastic interactions are discussed,which may facilitate the engineering of resistant crops.

Iron toxicity resistance strategies in tropical grasses:The role of apoplastic radicular barriers

The revegetation of mined areas poses a great challenge to the iron ore mining industry.The initial recovery process in degraded areas might rely on the use of Fe-resistant grasses.Tropical grasses, such as Paspalum densum and Echinochloa crus-galli, show different resistance strategies to iron toxicity; however, these mechanisms are poorly understood.The Fe-resistance mechanisms and direct iron toxicity as a function of root apex removal were investigated. To achieve this purpose, both grass species were grown for up to 480 hr in a nutrient solution containing 0.019 or 7 mmol/L Fe-EDTA after the root apices had been removed or maintained. Cultivation in the presence of excess iron-induced leaf bronzing and the formation of iron plaque on the root surfaces of both grass species, but was more significant on those plants whose root apex had been removed. Iron accumulation was higher in the roots, but reached phytotoxic levels in the aerial parts as well. It did not hinder the biosynthesis of chloroplastidic pigments. No significant changes in gas exchange and chlorophyll a fluorescence occurred in either grass when their roots were kept intact; the contrary was true for plants with excised root apices. In both studied grasses, the root apoplastic barriers had an important function in the restriction of iron translocation from the root to the aerial plant parts, especially in E. crus-galli. Root apex removal negatively influenced the iron toxicity resistance mechanisms(tolerance in P. densum and avoidance in E. crus-galli).

Editorial: Effect of root anatomy and apoplastic barrier development on cadmium uptake in rice

Cadmium (Cd) is a toxic metal with high mobility from soil and known translocation into plants (Song et al., 2015).Because the main source of human exposure to Cd is from food consumption, there has been increased research examining Cd uptake in agricultural plants (Li et al., 2014;Rizwan et al., 2016;Song et al., 2015).

A G-type lectin receptor-like kinase regulates the perception of oomycete apoplastic expansin-like proteins

Phytophthora capsici is one of the most harmful pathogens in agriculture, which threatens the safe production of multiple crops and causes serious economic losses worldwide. Here, we identified a P. capsici expansin-like protein, Pc EXLX1, by liquid chromatography-tandem mass spectrometry from Nicotiana benthamiana apoplastic fluid infected with P. capsici. Clustered regularly interspaced short palindromic repeats/crispr associated protein9(CRISPR/Cas9)-mediated Pc EXLX1 knockout mutants exhibited significantly enhanced virulence,while the overexpression of Pc EXLX1 impaired the virulence. Prokaryotically expressed Pc EXLX1 activated multiple plant immune responses, which were BRI1-associated kinase 1(BAK1)-and suppressor of BIR1-1(SOBIR1)-dependent. Furthermore, overexpression of Pc EXLX1 homologs in N. benthamiana could also increase plant resistance to P. capsici. A G-type lectin receptor-like kinase from N. benthamiana, expansin-regulating kinase 1(ERK1), was shown to regulate the perception of Pc EXLX1 and positively mediate the plant resistance to P. capsici. These results reveal that the expansin-like protein, Pc EXLX1, is a novel apoplastic effector with plant immunity-inducing activity of oomycetes, perception of which is regulated by the receptor-like kinase, ERK1.

Co-evolved plant and blast fungus ascorbate oxidases orchestrate the redox state of host apoplast to modulate rice immunity

Apoplastic ascorbate oxidases(AOs)play a critical role in reactive oxygen species(RoS)-mediated innate host immunity by regulating the apoplast redox state.To date,little is known about how apoplastic effectors of the riceblast fungus Magnaportheoryzaemodulate the apoplast redox state of rice to subvert plant immunity.In this study,we demonstrated that M.oryzae MoAo1 is an Ao that plays a role in virulence by modulating the apoplast redox status of rice cells.We showed that MoAo1 inhibits the activity of rice OsAO3and OsAO4,which also regulate the apoplast redox status and plant immunity.In addition,we found that MoAo1,OsAO3,andOsAO4 allexhibit polymorphic variations whosevaried interactions orchestrate pathogen virulence and rice immunity.Taken together,our results reveal a critical role for extracellular redox enzymes during rice blast infection and shed light on the importance of the apoplast redox state anditsregulation inplant-pathogeninteractions.

Ammonium improves iron nutrition by decreasing leaf apoplastic pH of sunflower plants(Helianthus annuus L.cv.Frankasol)

The effect of nitrogen form on pH and concentration of soluble iron (Fe) in leaf apoplast was investigated in hydrophonically grown sunflower plants (Helianthus annuus L. cv. Frankasol), and the mechanism underlying the improved Fe nutrition by ammonium (NH4) supply was also elucidated. Ammonium supply ameliorated Fe nutrition of plants grown without Fe through decreasing apoplastic pH and increasing soluble Fe concentration in apoplastic fluid of young leaves. The soluble Fe concentration in apoplastic fluid and cell sap of young leaves, and xylem exudates of NH4 fed-plants was higher than that of nitrate (NO3) fed-plants, and no typical Fe-deficiency chlorosis in young leaves was observed in NH4 fed plant without Fe supply. The apoplastic pH was 6.15 and 5.94 in young leaves of Fe-deficient plants fed respectively with NO3 and NH4, while in Fe-sufficient plants, the apoplastic pH was 6.43 with NO3, and 5.50 with NH4 supply. In primary leaves, the apoplastic pH was around 6.25 irrespective of nitrogen form and Fe supply. The pH of xylem exudate was 5.72 in Fe-deficient plants fed with NO3 and 5.49 with NH4. Iron nutrition increased the pH of xylem exudate by 0.27 and 0.16 unit under NO3 and NH4 supply respectively.

Anatomical and Chemical Alterations but not Photosynthetic Dynamics and Apoplastic Transport Changes are Involved in the Brittleness Culm Mutation of Rice

Brittleness culm is an important agronomic trait that has a potential usefulness in agricultural activity as animal forage although the developmental mechanism is not clear yet. In the present study,the anatomical and chemical characteristics as well as some ecophysiological features in the brittleness culm mutation of rice(Oryza sativa L.) were investigated. Compared with the wild type(WT),the brittleness culm mutant(bcm) exhibited higher culm vascular bundle distance and lower culm wall thickness,leaf interveinal distance and leaf thickness. Ratio of bundle sheath cell/whole bundle and areas of whole vascular bundles and bundle sheath of leaves were reduced while ratios of xylem and phloem to whole bundles were elevated in bcm. The Fourier transform infrared(FTIR) microspectroscopy analysis and further histochemical and physiological measurements revealed that the different contents and depositions of cell wall components such as pectins,lignin,suberin and cellulose all participated in the mutation of brittleness. However,the mutant presented no significant changes in leaf photosynthetic dynamics and apoplastic transport ability. These results strongly indicate that the alterations in anatomical and chemical characteristics,rather than changes in major ecophysiological features such as photosynthesis and apoplastic transport were involved in the brittleness mutation of rice.

冬青卫矛叶片质外体适应冬季胁迫的蛋白质组学分析

植物的质外体在感知外界信号和胁迫应答中起重要作用。该研究采用生理生化和蛋白质组学方法,对秋季和冬季冬青卫矛叶片的理化指标、微观结构以及叶片质外体液体中的蛋白变化进行比较分析,探索冬青卫矛叶片质外体响应冻胁迫的分子机制,以期为植物抗冻分子机制研究提供依据。结果表明:(1)冬季冬青卫矛叶片中MDA、可溶性糖含量以及SOD、POD活性均显著升高,气孔张开度减小,叶片厚度变小。(2)冬季冬青卫矛质外体液体中的蛋白质种类和含量显著高于秋季。(3)冬青卫矛叶片质外体液体中共鉴定到838个肽段和194个蛋白质;与秋季相比,冬季冬青卫矛叶片质外体液体中共筛选到43种差异积累蛋白(DAPs),其中26个蛋白质显著上调,17个蛋白质显著下调;蛋白表达模式显示,胚胎发育晚期丰富蛋白质、铁超氧化物歧化酶、过氧化物酶、丝氨酸羧肽酶等在冬季表达量较高,推测它们可能是冬季胁迫响应敏感的蛋白质。(4)KEGG富集分析显示,差异蛋白主要与应激防御、细胞壁修饰、抗病、自由基清除、甘油脂类代谢、淀粉和蔗糖代谢、次生代谢物的生物合成等生物学过程相关。(5)验证实验结果表明,冬季冬青卫矛8个差异积累蛋白与其对应的基因的表达趋势一致。研究认为,冬季冬青卫矛质外体液体中积累的蛋白可通过清除活性氧、促进单糖、寡糖和游离氨基酸等渗透调节物的生成而增强对环境的适应;推测冬青卫矛质外体中积累的单糖和寡糖可能通过增加质外体液体的浓度从而降低冰点,进而提高冬青卫矛对冬季胁迫的耐受性。

重金属镉(Cd)在植物体内的转运途径及其调控机制

重金属镉(Cd)的毒害效应与其由土壤向植物地上部分运输有关,揭示Cd^(2+)转运途径及其调控机制可为提高植物抗镉性以及镉污染的植物修复提供依据。对Cd^(2+)在植物体内的转运途径,特别是限制Cd^(2+)移动的细胞结构和分子调控机制研究进展进行了回顾。Cd^(2+)通过共质体和质外体途径穿过根部皮层进入木质部的过程中,大部分在皮层细胞间沉积,少部分抵达中柱后转移到地上部分。为了免受Cd^(2+)的危害,植物体产生了多种限制Cd^(2+)吸收和转移的生理生化机制:1)环绕在内皮层径向壁和横向壁上的凯氏带阻止Cd^(2+)以质外体途径进入木质部;2)螯合剂与进入根的Cd^(2+)螯合形成稳定化合物并区隔在液泡中;3)通过H+/Cd^(2+)离子通道等将Cd^(2+)逆向转运出根部。植物共质体和质外体途径转运重金属镉的能力以及两条途径的串扰尚待进一步明晰和阐明。

水稻胚乳的发育及其养分输入的途径

用光镜和电镜观察了水稻胚乳细胞及其中的淀粉体和蛋白质颗粒的发育过程;用整体解剖法查明了胚乳核的分裂方式,用微孔滤膜抽滤酶解细胞悬浮液的方法研究了胚乳细胞的增殖动态。主要结果如下:(1)水稻胚乳游离核与细胞的分裂有两种形式:无丝分裂与有丝分裂,并以无丝分裂为主。(2)核期结束时,游离核间形成成膜体,产生细胞壁。(3)胚乳细胞分裂的持续时间受到温度的影响,气温高时,胚乳细胞增殖快,在平均气温27℃时分裂的持续时间为12天。(4)淀粉体是由质体积累淀粉发育起来的,质体能通过长出芽胞或中间收缩断开的方式进行分裂。(5)胚乳细胞中有两种蛋白质颗粒,P_1与P_2,它们都在粗糙内质网上形成,P_1积累在由内质网围起的囊泡中,而P_2积累在液泡中。(6)由背部维管束卸出的灌浆物质必须经过质外体才能进入胚乳。糊粉层是胚乳吸收养分的细胞层。

植物根中质外体屏障结构和生理功能研究进展

本文综述了近10年来植物根中质外体屏障结构和功能的研究进展。质外体屏障指根中内、外皮层初生壁的凯氏带,或次生壁栓质化和木质化,以及植物体表角质层组成的保护组织,能隔绝水、离子和氧气不能自由进出植物体的屏障结构,具有保护植物体的生理功能。根中凯氏带的分子发育机理研究表明根内皮层类似哺乳动物上皮组织的保护作用。植物根中质外体保证内部各种生理代谢在稳定的内部环境中进行,是植物适应各种逆境的重要屏障结构。根中质外体屏障在植物适应干旱、洪涝灾害、离子胁迫和病虫害的侵袭等方面具有重要作用,在探索适应并修复极端生态环境的植物资源中有广阔的应用前景。

白茅解剖结构和屏障结构特征研究

利用光学显微镜和荧光显微镜对白茅进行了解剖学和组织化学研究,结果表明:1)白茅不定根解剖结构为表皮、外皮层、厚壁机械组织层、皮层、内皮层和中柱;根茎结构为角质层、表皮、周缘厚壁机械组织层、皮层、厚壁机械组织层和髓。2)不定根具内侧内皮层及其邻近皮层细胞、外侧表皮和皮下层组成的屏障结构;根茎具内侧厚壁机械组织层、外侧角质层和周缘厚壁机械组织层组成的屏障结构,屏障结构胞壁具凯氏带、木栓质和木质素沉积的组织化学特点。3)白茅通气组织包括根中通气组织,茎皮层通气组织和维管束中的气腔。4)白茅的屏障结构和解剖结构是其适应湿地环境的重要特征,但是根茎周缘厚壁机械组织层没有栓质化,髓部没有髓腔,推测其在湿地环境中分布有一定的局限性。

PEG胁迫下水稻根外皮层厚壁细胞的解剖学研究

通过PEG模拟干旱胁迫,采用Spurr树脂半薄切片结合显微观察的方法,比较水稻和旱稻根外皮层边缘厚壁细胞的形态差异,研究根外皮层边缘厚壁细胞加厚的规律。结果表明:随PEG浓度的升高,水稻(扬稻6号)根外皮层边缘厚壁细胞的胞壁宽度明显增加;旱稻(中旱3号)对低浓度PEG胁迫不太敏感,当PEG浓度达50g·L-1以上时,根外层厚壁细胞表现出明显加厚的趋势,且随PEG胁迫时间延长,厚壁细胞栓质化程度变高。推测水稻根系在干旱胁迫条件下具有一定的缓冲能力,外皮层边缘厚壁细胞在根外层导水率方面发挥显著的质外体屏障作用,可有效地防止水分丧失,保护根组织免受干旱的伤害。

天胡荽的解剖和屏障结构特征研究

利用光学显微镜和荧光显微镜对天胡荽进行了解剖学和组织化学研究,结果表明,1)天胡荽不定根为初生结构,具二原型维管柱、内皮层、皮层、外皮层和表皮。2)茎、花柄和叶主要为初生结构,除了茎和花柄维管束具次生结构,具表皮、厚角组织、皮层、内皮层、维管束和髓;茎具诱导型通气组织。3)天胡荽不定根的屏障结构包括内侧的有凯氏带且栓质化的内皮层,外侧的有凯氏带且栓质化的外皮层和紧邻外侧具扩散状栓质层的表皮细胞;匍匐茎、花柄和叶柄具相似的质外体屏障结构,一是内侧的有凯氏带且栓质化的内皮层,二是外侧的表皮外角质层,但花柄和叶柄有凯氏带的细胞,并不栓质化;叶片的仅为表皮外角质层。4)天胡荽的解剖和屏障结构特征是其适应多种水湿环境的结构基础。该研究为今后选择湿地生态修复植物提供线索。

2种杨树嫩茎质外体内源激素对盐胁迫的响应

以春季盆栽的‘吴屯杨’与‘荷兰杨’为试验材料,采用微透析技术,结合高效液相色谱测定不同浓度NaCl(0、200、300mmol/L)胁迫下2种杨树嫩茎质外体的内源激素变化.结果表明:盐胁迫下2种杨树嫩茎质外体内脱落酸(ABA)含量升高,其他激素含量降低;高浓度盐胁迫下脱落酸(ABA)的积累量大于中浓度盐胁迫;同等盐胁迫条件下,‘荷兰杨’需要积累较‘吴屯杨’更多的脱落酸(ABA)来抵御胁迫环境;‘吴屯杨’的抗盐平衡系数小于‘荷兰杨’;2种杨树的细胞分裂素(CTK)与吲哚乙酸(IAA),细胞分裂素(CTK)与赤霉素(GA),吲哚乙酸(IAA)与赤霉素(GA)显著相关;‘吴屯杨’的耐盐性强于‘荷兰杨’.

微透析法获取鹅掌柴嫩茎质外体汁液的研究

该文研究的重点是获取质外体汁液。假设微透析技术可以在植物上得到质外体汁液,并以盆栽鹅掌柴为材料,在活体嫩茎插入微透析探针后的不同时间段内分别收集透析液,测定其中的离子浓度和苹果酸脱氢酶活性加以验证。结果表明:在探针插入120 min后,收集的透析液中Na+、K+和Ca2+浓度逐渐趋于平缓,并且苹果酸脱氢酶的活性消失,从而证明120 min后得到的微透析液,没有受到破损细胞液的污染,是纯净的鹅掌柴嫩茎质外体汁液,从而为活体、方便、快捷地获取植物质外体汁液提供新的方法。