甜菜细胞壁界面特征显著影响纤维素酶解效率

生物质不同器官、组织理化性质各异,这种异质性对纤维素酶解效率的影响有待深入研究。该研究以甜菜为研究对象,系统分析不同器官理化性质与酶解效率间的关系,探究细胞壁与纤维素酶的吸附特点及在酶解过程中的形态变化。结果表明,甜菜根、茎、叶酶解还原糖得率不同,根和茎的还原糖得率相近且高于叶,分别为14.64%和14.26%,叶还原糖得率较低为10.15%;酶解还原糖得率与木质素（P<0.01）、半纤维素（P<0.01）呈极显著负相关,而与比表面积（P<0.01）呈极显著正相关;甜菜根、茎、叶的薄壁组织更容易与纤维素酶结合,并且更易被降解。甜菜细胞壁界面理化特征是影响酶与底物有效吸附,进而影响纤维素酶解效率的关键因素。研究结果可为甜菜农业废弃物的利用提供参考。     

Cell wall polysaccharide chemistry of peach genotypes with contrasted textures and other fruit traits

Cell wall composition, pectin, and hemicellulose fine structure variation were assessed in peach and related genotypes with contrasted texture and fruit shape. Cell walls were prepared from four commercial peaches, eight genotypes from the Jalousia × Fantasia peach cross, and six genotypes from the Earlygold peach × Texas almond cross. Sugar composition was determined chemically while fine structure of homogalacturonan pectin and xyloglucan hemicellulose were assessed by coupling pectin lyase and glucanase degradation, respectively, with MALDI-TOF MS analysis of the degradation products. The results indicate clear compositional and structural differences between the parents and their related genotypes on the basis of pectin versus cellulose/hemicellulose content and on the fine structure of homogalacturonan and xyloglucan. A relation between methyl- and acetyl-esterification of pectin with fruit shape is revealed in the Fantasia × Jalousia peach genotypes.     

Moderate ferulate and diferulate levels do not impede maize cell wall degradation by human intestinal microbiota

The degradation of plant fiber by human gut microbiota could be restricted by xylan substitution and cross-linking by ferulate and diferulates, for example, by hindering the association of enzymes such as xylanases with their substrates. To test the influence of feruloylation on cell wall degradability by human intestinal microbiota, nonlignified primary cell walls from maize cell suspensions, containing various degrees of ferulate substitution and diferulate cross-linking, were incubated in nylon bags in vitro with human fecal microbiota. Degradation rates were determined gravimetrically, and the cell walls were analyzed for carbohydrates, ferulate monomers, dehydrodiferulates, dehydrotriferulates, and other minor phenolic constituents. Shifting cell wall concentrations of total ferulates from 1.5 to 15.8 mg/g and those of diferulates from 0.8 to 2.6 mg/g did not alter the release of carbohydrates or the overall degradation of cell walls. After 24 h of fermentation, the degradation of xylans and pectins exceeded 90%, whereas cellulose remained undegraded. The results indicate that low to moderate levels of ferulates and diferulates do not interfere with hydrolysis of nonlignified cell walls by human gut microbiota.     

Apoplastic pH and monolignol addition rate effects on lignin formation and cell wall degradability in maize

Monolignol polymerization rate and apoplastic pH and may influence the formation of lignin and its interactions in cell walls. Primary maize walls were artificially lignified by gradual "end-wise" or rapid "bulk" polymerization of coniferyl alcohol at pH 4 or 5.5. Lignification efficiency was greatest for end-wise polymers at pH 5.5 (90-98%), intermediate for bulk polymers formed at either pH (54-82%), and lowest for end-wise polymers at pH 4 (41-53%). End-wise polymers had about 2.2-fold more ether inter-unit linkages and 70% fewer end-groups than bulk polymers. Low pH enhanced the formation of ether linkages in end-wise but not in bulk polymers. Differences in lignin structure did not influence the enzymatic degradability of cell walls, but lowering apoplastic pH from 5.5 to 4.0 during lignification reduced cell wall degradability by 25%. Further studies indicated this pH-dependent depression in degradability was related to cell wall cross-links formed via lignin quinone methide intermediates.     

Effect of phenolic structures on the degradability of cell walls isolated from newly extended apical internode of tall fescue (Festuca arundinacea Schreb.)

Apical internodes of tall fescue (Festuca arundinacea Schreb. var. Clarine) harvested at flowering were sectioned into 5 or 10 equal parts to study in situ degradability and cell wall composition, respectively. The basal (youngest) section had the greatest primary wall content. Cell walls in the upper (older) sections had the highest xylose/arabinose ratio and lignin content and a lignin rich in syringyl units, all typical of extensive secondary wall development. Almost all of the p-coumaric (p-CA) and about half of the ferulic acid (FA) were released by 1 M NaOH and presumed to be ester-linked. The total FA content was approximately double that of p-CA in all sections other than the youngest with a distribution similar to that of total p-CA. However, the ratio of esterified to ether and ether plus ester linked (Et & Et+Es) FA differed with age. Whereas the esterified form remained essentially constant ( approximately 4.5 g/kg of cell wall), Et & Et+Es ferulate increased with increasing age of the tissue and was significantly related to lignin deposition (r = 0.79, P < 0.01). The extent of cell wall degradation after 48 h of incubation in the rumen was inversely related to maturity, falling from 835 g/kg of dry matter in the youngest section to 396 g/kg in the oldest. Both the rate and extent of cell wall degradation were significantly negatively related to the ratio of xylose to arabinose, lignin content, proportion of syringyl units present in lignin, and concentration of Et & Et+Es FA present. A positive relationship between Et & Et+Es FA was also found, with the rate (P < 0.01) being better correlated than the extent (P < 0.05) of cell wall degradation. Application of the newly extended internode model to fescue produced results consistent with the view that both the lignin content and the extent to which lignin was covalently bound to the other wall polymers crucially influenced the rate and extent of degradation.     

Methyl esterification divergently affects the degradability of pectic uronosyls in nonlignified and lignified maize cell walls

Nonlignified cell walls from Zea mays (L.) cell suspensions were incubated with and without pectin methylesterase (PME) and a portion were artificially lignified to assess how methyl esters influence the release of pectic uronosyls and total sugars from cell walls by fungal enzymes. Treatment with PME reduced uronosyl concentrations from 97 to 92 mg/g, reduced uronosyl methylation from 57% to 21%, and increased Klason lignin concentrations in artificially lignified cell walls from 99 to 116 mg/g. Although PME treatment slightly enhanced uronosyl release from nonlignified cell walls, it reduced uronosyl release from artificially lignified cell walls by 55% after 4 h and by 7% after 72 h of enzymatic hydrolysis. Pectin hydrolysis in PME treated cell walls was probably impaired by enhanced benzyl ester cross-linking of uronosyls to lignin via quinone methide intermediates. Variations in uronosyl methylation had little effect on the overall release of total sugars from cell walls.     

铝胁迫下植物根尖细胞壁组成物质变化抑制根伸长的生理机制研究

根伸长受抑制是植物受铝毒害的主要症状,铝诱导的细胞壁组成物质的变化是其主要原因。本文主要对铝胁迫下植物根尖细胞壁组成物质如木质素、 胼胝质、 纤维素、 半纤维素、 果胶、 细胞壁多糖蛋白及相关代谢酶类在铝胁迫下的变化对根伸长的影响及生理机理的研究进展进行了综述,明确了铝胁迫诱导的植物根尖细胞壁组成物质含量、 比例及结构的变化导致细胞壁刚性降低,从而抑制细胞伸长,最终抑制根伸长。本文还指出,鉴于缺乏对同一植物甚至同一个种类的植物根尖细胞壁各主要组成物质铝胁迫下变化的系统研究,不能对造成该植物根伸长受抑制的原因做出全面合理的解释,所以今后应侧重于铝胁迫下各细胞壁组分变化在抑制根伸长中的贡献率的研究,尤其要针对主要粮食作物进行系统研究,以有效解决铝胁迫造成的产质量降低。     

Impact of lignin structure and cell wall reticulation on maize cell wall degradability

In this study, eight maize recombinant inbred lines were selected to assess both the impact of lignin structure and the impact of cell wall reticulation by p-hydroxycinnamic acids on cell wall degradability independently of the main "lignin content" factor. These recombinant lines and their parents were analyzed for in vitro degradability, cell wall residue content, esterified and etherified p-hydroxycinnamic acid content, and lignin content and structure. Lignin structure and esterified p-coumaric acid content showed significantly high correlation with in vitro degradability (r=-0.82 and r=-0.72, respectively). A multiple regression analysis showed that more than 80% of cell wall degradability variations within these 10 lines (eight recombinant inbred lines and their two parents) were explained by a regression model including two main explanatory factors: lignin content and estimated proportion of syringyl lignin units esterified by p-coumaric acid. This study revealed new biochemical parameters of interest to improve cell wall degradability and promote lignocellulose valorization.     

NaOH预处理提高甘蔗叶产甲烷性能及其机理分析

为提高甘蔗叶厌氧消化的产甲烷性能,采用NaOH对粉碎后的甘蔗叶进行了预处理,得到了不同NaOH浓度、不同预处理时间条件下甘蔗叶厌氧消化的甲烷产率,并研究比较了预处理前后甘蔗叶微观物理形态、化学分子结构和化学组分的变化。结果表明:与未预处理甘蔗叶相比,NaOH预处理甘蔗叶的累计产甲烷量提高了22.02%～89.94%,厌氧消化时间T80缩短了2～4 d,其中6%NaOH-5d预处理甘蔗叶的产甲烷性能最好;NaOH破坏了甘蔗叶表面蜡质层和细胞壁结构,促进了甘蔗叶表面二氧化硅、木质素等分解,打破了对纤维素的束缚;预处理后甘蔗叶的木质纤维素结构发生明显变化,其中木质素的羟基、甲氧基和羰基等部分官能团发生不同程度断裂,紧致的大分子结构发生分解,纤维素的结晶度降低,部分氢键遭到破坏,半纤维素发生了分子间和分子内的降解;预处理甘蔗叶的木质纤维素含量均有不同程度的降低,可被微生物分解利用的有机物质增多,其中6%NaOH-5d预处理甘蔗叶厌氧消化的木质素、纤维素、半纤维素降解率分别提高了9.27%、25.14%和21.52%。因此NaOH预处理是一种提高甘蔗叶厌氧消化产甲烷性能的有效方法。     

Solid-state selective (13)C excitation and spin diffusion NMR to resolve spatial dimensions in plant cell walls

The average spatial dimensions between major biopolymers within the plant cell wall can be resolved using a solid-state NMR technique referred to as a (13)C cross-polarization (CP) SELDOM (selectively by destruction of magnetization) with a mixing time delay for spin diffusion. Selective excitation of specific aromatic lignin carbons indicates that lignin is in close proximity to hemicellulose followed by amorphous and finally crystalline cellulose. (13)C spin diffusion time constants (T(SD)) were extracted using a two-site spin diffusion theory developed for (13)C nuclei under magic angle spinning (MAS) conditions. These time constants were then used to calculate an average lower-limit spin diffusion length between chemical groups within the plant cell wall. The results on untreated (13)C enriched corn stover stem reveal that the lignin carbons are, on average, located at distances ～0.7-2.0 nm from the carbons in hemicellulose and cellulose, whereas the pretreated material had larger separations.     

Distribution of Accumulated Aluminum and Changes in Cell Wall Polysaccharides in Eucalyptus camaldulensis and Melaleuca cajuputi under Aluminum Stress

Distribution of aluminum (Al) within plant components and Al-induced changes in cell wall polysaccharides in root tips of Eucalyptus camaldulensis Dehnh. seedlings were compared with those of Melaleuca cajuputi Powell. In E. camaldulensis , 0.5 mM Al (pH 4.2 for 40 d) reduced plant dry weight by 50%, increased callose concentration in the root tips and induced leaf necrosis. In comparison with M. cajuputi , Al concentrations were higher in roots and leaves of E. camaldulensis on both a fresh weight basis and in the cell sap, but were lower in the cell wall. Al increased pectin, hemicellulose and cellulose concentration in the cell walls of E. camaldulensis root tips. Al-induced leaf necrosis and growth reduction in E. camaldulensis is discussed in the context of potentially toxic concentrations of Al in plant tissue and changes in polysaccharide content which could reduce water and nutrient uptake and cell wall extensibility in roots.     

Studies on the metabolism in roots of lowland rice

In the previous studies we investigated some characteristics of paddy root in comparison with shoot by analysis of inorganic elements¹⁾ and metabolizable forms of organic compounds²⁾ of shoot and root at successive growth stages, and found that the contents of the above-mentioned components in root were generally lower than those in shoot with few exceptions. From this it is concluded that in root the contents of the residual components, mainly cell wall substances, must be high. To ascertain this surmise, we carried out analysis of shoot, root and inflorescences of rice plant with respect to hemicellulose, cellulose and lignin which construct cell walls.     

Tissue-specific and developmental modifications of grape cell walls influence the adsorption of proanthocyanidins

Cell wall material from Vitis vinifera L. cv. Cabernet Sauvignon grape skin and flesh was isolated at different stages of grape maturity to determine whether developmental changes in cell wall composition in different tissue types influence the binding of proanthocyanidins (PAs). Trends in cell wall adsorption of, and selectivity for, PAs were determined using two skin PAs that differed in their average molecular masses. Flesh cell walls consistently bound a higher amount of PA than those from skin. Key structural differences that reduced PA adsorption in skin cell walls by comparison with flesh cell walls were endogenously higher concentrations of insoluble PA, Klason lignin, and lower cell wall-bound protein. These differences may confer reduced flexibility and porosity of skin cell walls relative to flesh cell walls. Analysis of skin and flesh cell wall properties revealed that the onset of ripening was associated with a loss of type I arabinogalactan and galacturonic acid, which indicated a degradation of pectin within the cell wall. Flesh cell walls consistently bound PAs of larger molecular mass, and changes in PA adsorption properties after the onset of ripening were minor. For skin cell walls, adsorption of PA was lowest immediately following solubilization of galacturonic acid, and high molecular mass PAs were poorly bound. As ripening progressed, PAs of higher molecular mass were selectively adsorbed by skin cell walls, which indicates that ongoing cell wall remodeling during ripening may confer an increased porosity within the skin cell wall matrix, resulting in a greater adsorption of PA within a permeable structure.     

Temporal sequence of cell wall disassembly events in developing fruits. 2. Analysis of blueberry (Vaccinium species)

Softening and pathogen susceptibility are the major factors limiting the marketing of blueberries as fresh fruits, and these traits are associated with fruit cell wall structure. However, few studies that characterize wall modifications occurring during development and ripening have been reported for this fruit. In this study the ripening-associated modifications of blueberry fruit cell walls (composition, pectin and hemicellulose solubilization, and depolymerization) at five stages of ripeness have been analyzed. Xylose was found to be the most abundant noncellulosic neutral sugar associated with fruit walls, and the observed high Xyl/Glc ratio suggested that xylans, which are usually a minor hemicellulosic fruit wall component, are abundant in blueberry. The pectic matrix showed increased solubilization at early and intermediate stages of ripening, but no changes were detected in late ripening. Furthermore, little reduction in pectin polymer size occurred during blueberry ripening. In contrast, hemicellulose levels decreased as ripening progressed, and a clear depolymerization of these components was observed. A model for cell wall degradation in this fruit is discussed.     

Distribution of Accumulated Aluminum and Changes in Cell Wall Polysaccharides in Eucalyptus camaldulensis and Melaleuca cajuputi under Aluminum Stress

Distribution of aluminum (Al) within plant components and Al-induced changes in cell wall polysaccharides in root tips of Eucalyptus camaldulensis Dehnh. seedlings were compared with those of Melaleuca cajuputi Powell. In E. camaldulensis, 0.5 mM Al (pH 4.2 for 40 d) reduced plant dry weight by 50%, increased callose concentration in the root tips and induced leaf necrosis. In comparison with M. cajuputi, Al concentrations were higher in roots and leaves of E. camaldulensis on both a fresh weight basis and in the cell sap, but were lower in the cell wall. Al increased pectin, hemicellulose and cellulose concentration in the cell walls of E. camaldulensis root tips. Al-induced leaf necrosis and growth reduction in E. camaldulensis is discussed in the context of potentially toxic concentrations of Al in plant tissue and changes in polysaccharide content which could reduce water and nutrient uptake and cell wall extensibility in roots.     

小麦根尖细胞壁对铝的吸附/解吸特性及其与耐铝性的关系

研究了耐铝性明显差异的2个小麦基因型西矮麦1号(耐性)和辐84系(敏感)根系对铝毒胁迫的反应与根尖细胞壁组分以及细胞壁对铝的吸附和解吸的关系。结果表明,30mol/L.AlCl3可迅速抑制小麦根系伸长,但对辐84系根系伸长的抑制更为明显,且小麦根系相对伸长率随着铝浓度的提高而急剧降低。在30mol/L.AlCl3处理24h后,西矮麦1号根系伸长的抑制率为33.3%,而辐84系根系伸长的抑制率高达70.9%。小麦距根尖0～10.mm根段的铝含量和细胞壁中果胶糖醛酸含量显著高于10～20.mm根段,且前者对铝的累积吸附量明显大于后者;在0～10.mm根段,敏感基因型果胶含量高于耐性基因型,其根尖含铝量及根尖细胞壁对铝的吸附量都要大于后者。采用1.0.mol/L.NH3.H2O对细胞壁预处理2.h降低果胶甲基酯化程度后,耐性和敏感基因型根尖细胞壁对铝的累积吸附量分别降低了17.1%和20.9%,但对铝的累积解吸率没有影响。由此可见,小麦根尖是铝毒的主要位点,细胞壁果胶含量和果胶甲基酯化程度可能是导致不同小麦基因型根尖细胞壁对铝吸附量、铝积累量的差异及其对铝毒胁迫反应的差异的重要原因。     

镉安全水稻材料根系细胞壁果胶对镉的响应特征

【目的】研究镉(Cd)处理下水稻根系细胞壁果胶对Cd胁迫的响应,进一步深化Cd安全水稻材料根系细胞壁Cd的固持机制。【方法】以Cd安全水稻材料D62B为研究对象,普通材料Luhui17为对照进行水培试验。设4个Cd质量浓度处理：0 mg/L (CK)、0.5 mg/L (Cd0.5)、1.0 mg/L (Cd1)、2.0 mg/L (Cd2)。在水稻分蘖期采集根系样品,分析细胞壁多糖中果胶、半纤维1、半纤维2以及残渣部分的Cd含量,测定果胶糖醛酸含量、果胶酯化度、果胶甲酯酶(PME)活性、根系过氧化氢(H₂O₂)含量以及细胞壁过氧化物酶(POD)活性,进而分析根系细胞壁果胶对Cd的响应特征。【结果】1) Cd胁迫下,D62B和Luhui17根系细胞壁果胶合成增加,根系细胞壁低酯化和高酯化果胶糖醛酸含量均表现为D62B高于Luhui17。Cd处理下D62B根系细胞壁低酯化和高酯化果胶糖醛酸含量较对照分别增加了13.21%～71.82%和22.10%～64.27%,Luhui17分别增加了24.14%～137.86%和13.12%～41.26%。...     

猕猴桃果实贮藏期间细胞壁多糖物质降解特性及组织结构差异分析

为了深入了解猕猴桃果实采后细胞壁多糖物质降解及组织结构变化与果实贮藏性的关系,本研究以红阳和华特猕猴桃果实为试材,对25和4℃贮藏期间的细胞壁多糖物质含量及果胶降解酶活性进行测定,并比较两品种果实在25℃贮藏期间的细胞显微结构和钙组分含量差异。结果表明,在25和4℃贮藏条件下,随着贮藏时间的延长,红阳和华特猕猴桃果实中半纤维素（HCL）、纤维素（CL）和共价结合型果胶（CSP）含量不断降低,水溶性果胶（WSP）含量不断上升,离子结合型果胶（ISP）含量相对稳定。红阳猕猴桃各细胞壁多糖组分含量变化速度较快。两品种猕猴桃果实硬度均与WSP含量呈显著负相关,与CSP含量呈显著正相关。果胶降解酶活性检测结果显示,25℃贮藏前期,红阳猕猴桃中果胶酸裂解酶（PL）和β-半乳糖苷酶（β-Gal）活性显著高于华特;贮藏中后期,红阳中果胶甲酯酶（PME）活性显著高于华特。4℃贮藏期间,红阳中PME活性仍显著高于华特;4℃贮藏前期,红阳中β-Gal活性与华特无显著差异,而PL活性低于华特。相关性分析表明,25℃贮藏期间,与红阳和华特果实软化显著相关的果胶降解酶分别是PME和PL;4℃贮藏期间,与华特果实软...     

硼对豌豆根尖细胞壁组分对铝吸附解吸的影响

  【目的】  比较加硼和不加硼条件下豌豆根尖细胞壁组分对铝的吸附解吸特征的差异,探讨硼对植物铝胁迫的缓解机制。  【方法】  以中豌6号豌豆(Pisum Sativum)为试验材料,在硼(0.6 μmol/L)水平下水培6天,提取豌豆根尖1 cm段细胞壁各组分并进行铝吸附解吸试验,根尖细胞壁各组分分别为螯合态果胶(果胶1),碱溶态果胶(果胶2),半纤维素和纤维素,并分析3个不同硼处理 (无硼、50 μmol/L H₃BO₃和 50 μmol/L 3-硝基苯硼酸) 条件下各组分对铝吸附解吸的影响。  【结果】  豌豆根尖细胞壁各组分含量为:纤维素>半纤维素>果胶2>果胶1。硼能够与果胶1和果胶2发生络合反应,与半纤维素也可能发生络合反应,从而影响果胶1、果胶2和半纤维素对铝的吸附解吸。在铝胁迫下,根尖细胞壁中的果胶是主要的铝结合位点,以果胶2结合最多。与对照比,硼处理显著提高了果胶2对铝的吸附量,但解吸量变化不显著。pH 3.5条件下,硼酸与3-硝基苯硼酸处理相比,更能有效地影响果胶对铝的吸附解吸。因此将铝固定在细胞壁的果胶2内,可能是硼酸缓解铝毒的重要机制之一。  【结论】  细胞壁是铝的主要结合部位,细胞壁果胶2是根尖细胞壁主要的铝结合位点,硼与果胶2的结合可能是硼缓解铝毒的重要机制之一。     

Fractionation of flax shives by water and aqueous ammonia treatment in a pressurized low-polarity water extractor

Fractionation of flax shives into cellulose, hemicellulose, and lignin with a two-stage extraction process using water and aqueous ammonia was carried out in a pressurized low-polarity water extractor operated at different temperatures, flow rates, and ammonia concentrations. During the first stage with water, 84% of hemicellulose and 32% of lignin were removed at 190 degrees C at a flow rate of 1.5 mL/min for 30 min. During the second stage with aqueous ammonia, more than 77% of the lignin was removed, and hemicellulose removal reached 95% at 200 degrees C at a flow rate of 0.5 mL/min and with a solvent/feed ratio of 40 mL/g. The temperature and flow rate had a significant effect on lignin removal. The impact of additives (anthraquinone and hydrogen peroxide) and modifications (overnight soaking, reduced particle size, and elevated temperature) on lignin extraction was also studied. The combination of higher temperatures and reduced particle sizes resulted in enhanced lignin extraction. The extraction profiles of free phenolics (vanillin, acetovanillone, and vanillic acid) during the two-stage processing were monitored and compared with those of lignin.