苹果果实发育期间细胞壁组分变化特性

以 '富士'、'国光'、'红星'、'金冠'和'嘎拉'5个苹果品种为试材,分析了果实发育成熟过程细胞壁物质(CWM)、水溶性果胶(WSP)、共价结合果胶(CSP)、离子结合果胶(ISP)、纤维素及半纤维素各组分变化.结果表明:在苹果果实发育过程中,5个品种果实CWM含量变化均呈先升后降的变化规律,均以果实膨大期为其含量下降的转折点;果实总果胶含量均呈不断降低的趋势,其中CSP为主导成分,'富士'和'国光'果实CSP含量最高,WSP含量最低,'嘎拉'与'红星'果实3种果胶含量变化居中,'金冠'果实总果胶含量最低且变化小,但在近成熟期'红星'和'金冠'果实WSP呈明显的上升趋势.果实半纤维素含量也具相似的变化规律,'国光'、'富士'和'金冠'等3个品种的高峰值显著高于'嘎拉'和'红星';比较5个品种纤维素含量,'国光'果实在成熟期之前显著高于其他4品种,而其他4品种的纤维素含量变化比较平稳.     

枇杷冷藏过程中果肉木质化与细胞壁物质变化的关系

“大红袍”和“解放钟”枇杷果实在 1℃下贮藏时 ,细胞壁物质代谢异常 ,果肉硬度持续升高而出汁率逐渐降低 ,果胶酯酶 (PE)和多聚半乳糖醛酸酶 (PG)活性和水溶性果胶含量下降 ,原果胶含量、苯丙氨酸解氨酶(PAL)活性及木质素和纤维素含量不断增加。约经 3周贮藏后 ,果实出现果皮难剥、果肉质地变硬、粗糙少汁的异常劣变现象。在 12℃下贮藏的枇杷果实 ,细胞壁物质代谢正常 ,果肉硬度增加少 ,PE和PG活性及水溶性果胶含量较高 ,无原果胶增加现象 ,PAL活性呈下降趋势 ,木质素和纤维素含量变化不大 ,果实不出现木质化败坏。这些结果表明冷藏枇杷的木质化败坏可能是一种低温失调现象     

枇杷冷藏过程中果肉木质化与细胞壁物质变化的关系

“大红袍”和“解放钟”枇杷果实在１℃下贮藏时,细胞壁物质代谢异常,果肉硬度升高而出汁率逐渐降低,果胶酯酶（ＰＥ）和多聚半乳糖醛酸酶（ＰＧ）活笥和水溶性果胶含量下降,原果胶含量、苯丙氨酸解氨酶（ＰＡＬ）活性及木质素和纤维含量不断增加。约经３周贮藏后,果实出现果皮难剥、果肉质地变硬、粗糙少汁的异常劣变现象。在１２℃下贮藏的枇杷果实,细胞壁物质代谢正常,果肉硬度增加少,ＰＥ和ＰＧ活性及水溶性果胶含量较遍     

香蕉果实成熟软化时果皮和果肉中α-L-阿拉伯呋喃糖苷酶活性的变化

阿拉伯糖是果实软化过程中变化最明显的细胞壁糖残基之一,α-L-阿拉伯呋喃糖苷酶是导致细胞壁多糖中阿拉伯糖残基降解的主要糖苷酶。为阐明该酶在香蕉果实成熟软化中的作用,实验对香蕉贮藏过程中果皮和果肉中该酶活性以及果实硬度、呼吸强度和乙烯释放量的变化进行了研究。结果表明:α-L-阿拉伯呋喃糖苷酶在果实初期的变化很小,到果实硬度开始急剧下降时达到最大,增加量达10倍以上,且果肉中的酶活性大于果皮中;乙烯吸收剂处理延缓了香蕉果实呼吸和乙烯高峰的出现时间,降低了果实硬度、果皮和果肉中α-L-阿拉伯呋喃糖苷酶活性变化的速度和幅度。以上结果表明α-L-阿拉伯呋喃糖苷酶起诱导香蕉果实成熟的作用,在果实的软化中起着十分重要的作用,且其活性受乙烯的调节。     

1-甲基环丙烯处理对采后杨桃果实软化和细胞壁代谢的影响

为探讨1-甲基环丙烯(1-methylcyclopropene, 1-MCP)延缓采后杨桃果实软化的作用机理,本文研究了0.6 μL/L 1-MCP处理对在(15±1)℃、相对湿度90%下贮藏的‘香蜜’甜杨桃(Averrhoa carambola Linn. cv. Xiangmi)果实软化和细胞壁代谢的影响。结果表明：与对照果实相比,1-MCP处理可保持较高的杨桃果实硬度,有效降低果胶酯酶(pectinesterase, PE)、多聚半乳糖醛酸酶(polygalacturonase, PG)、纤维素酶等细胞壁降解酶活性,延缓原果胶、纤维素、半纤维素含量的下降和水溶性果胶含量的增加。因此认为,0.6 μL/L 1-MCP处理能有效控制采后‘香蜜’甜杨桃果实的软化进程,延长果实保鲜期。     

贮藏期柚粒化过程中果实总细胞壁物质积累和维管束超微结构的变化

汁胞粒化是柑橘类果实一类普遍的生理失调病害,主要表现为汁胞硬度增加,果实品质降低。为了明确汁胞粒化过程其他果实组织的生理代谢特征,该试验以成熟‘琯溪蜜柚’果实为材料,室温贮藏60 d,测定不同贮藏阶段果实背面维管束汁胞、侧面维管束汁胞、囊衣和果皮总细胞壁物质含量,以及两类汁胞可溶性固形物含量,同时利用透射电子显微镜观察果实背面维管束和侧面维管束细胞超微结构的动态变化。结果显示：(1)贮藏10 d时两类果实维管束的筛管和伴胞次生细胞壁开始明显加厚,韧皮部薄壁细胞线粒体和囊泡数量开始增多,而且次生细胞壁也开始明显加厚;贮藏20 d时两类维管束韧皮部薄壁细胞线粒体和囊泡数量持续增加,而且高尔基体出现(之后消失),同时囊衣和果皮总细胞壁物质含量开始显著提高;贮藏40 d时仅侧面维管束韧皮部薄壁细胞线粒体数量持续增多,侧面维管束汁胞总细胞壁物质含量开始显著升高;贮藏60 d时两类果实维管束次生细胞壁持续加厚,囊衣、果皮和侧面维管束汁胞总细胞壁物质含量均持续显著升高,然而至贮藏期结束背面维管束汁胞总细胞壁物质含量始终无显著变化。(2)贮藏期内囊衣总细胞壁物质含量始终显著高于果皮,而果皮总细胞壁物质含量始终显著高于两类汁胞;贮藏后期侧面维管束汁胞总细胞壁物质含量显著高于背面维管束汁胞。(3)在果实贮藏过程中背面维管束汁胞可溶性固形物含量始终无显著变化,而侧面维管束汁胞可溶性固形物含量从贮藏40 d至贮藏期结束持续显著降低。研究表明,贮藏期柚果实维管束、囊衣和果皮中细胞壁物质代谢的变化早于汁胞;发现果实维管束韧皮部薄壁细胞内线粒体数量增加的同时维管束次生细胞壁明显加厚,在整个贮藏期内侧面维管束汁胞可溶性固形物含量的显著降低伴随着总细胞壁物质含量的显著升高。这些结果可能有助于柑橘类果实粒化机理的全面揭示。     

香蕉果实后熟过程中果肉软化差异的研究

对香蕉果实贮藏过程中内、外果肉相关生理生化指标及细胞结构的变化进行系统的观察分析,结果显示:(1)在贮藏初期香蕉果实内果肉的硬度小于外果肉,在贮藏过程中的同一时期,均表现出内果肉硬度小于外果肉,且内果肉硬度较外果肉先降到零;(2)在贮藏初期内果肉中多聚乳糖醛酸酶(PG)和淀粉酶的活性均高于外果肉,随着贮藏时间的延长,酶活性在内、外果肉均表现出不断增加,且这两种酶活性在内果肉中早于外果肉达到最高值,但其在内果肉中的最高值均略低于外果肉的最高值;而淀粉含量却相反,在贮藏初期内果肉中淀粉含量低于外果肉,且在贮藏过程中的降解速率高于外果肉;(3)超微结构显示,香蕉果实内果肉中淀粉粒和细胞壁结构的降解均早于外果肉.研究表明,香蕉果实的软化首先由内果肉细胞降解开始,并呈放射状向外逐步延伸.     

赤霉素和萘乙酸对柿果实采后成熟软化生理指标的影响

以采后'富平尖柿'果实为试材,常温下用60 mg/L赤霉素(GA3)和20 mg/L α-萘乙酸(NAA)进行处理,考察柿果实成熟软化相关生理指标及果胶物质代谢在贮藏过程中的变化,以明确GA3和NAA处理对柿果实贮藏效果的影响.结果显示:GA3、NAA处理果实的贮藏时间分别比对照延长了4 d和10 d;GA3和NAA处理可显著延缓果实硬度的下降进程,有效降低果实呼吸强度和乙烯释放量,且呼吸高峰和乙烯高峰的出现明显迟于对照;果实多聚半乳糖醛酸酶(PG)活性的升高受到抑制,从而延缓了原果胶的降解以及水溶性果胶含量的增加,阻碍了果实的软化进程.试验表明,GA3和NAA处理可有效延缓柿果实的后熟软化,延长其贮藏期限, 并以GA3的效果尤为明显.     

桃果实采后贮藏过程中细胞壁多糖的变化

以‘雨花三号’水蜜桃果实为试材,分别在5℃（低温）和20℃（常温）贮藏一段时间后,研究桃果实采后细胞壁多糖降解、硬度以及乙烯释放速率的变化特征。结果表明,乙烯释放高峰明显滞后于果实采后硬度的快速下降期。不同温度下贮藏过程中果实细胞壁多糖变化的对比表明,低温抑制了细胞壁果胶和细胞壁其余组分的变化,从而抑制了果实的软化。富含半乳糖醛酸的果胶主链断裂。果胶和细胞壁其余组分也发生了半乳糖和阿拉伯糖等中性糖的损失,说明果胶和细胞壁其余组分的增溶及其侧链中性糖的降解也是桃果实采后软化的重要因素,这可能是由多种相关多糖降解酶的作用所导致的。但半纤维素多糖中中性糖的降解对桃果实采后软化的进程并没有影响。     

pH值、温度和金属离子对endo-PG降解香蕉果胶多糖的影响

从香蕉果肉中分离出水溶性果胶多糖(WSP)和酸溶性果胶多糖(ASP),探讨了pH值、温度、金属离子对内切多聚半乳糖醛酸酶(endo-PG)降解果胶多糖的影响.结果表明,pH 3.7～4.6较适于endo-PG的作用.在相同pH值下,endo-PG对WSP的降解效应随香蕉果实的成熟逐渐增强,而对ASP则逐渐减弱;在20～40℃内,温度越高,endo-PG对WSP和ASP的降解作用越强.Cd~(2+)、Cu~(2+)、Mg~(2+)和Fe~(3+)可显著抑制endo-PG对果胶多糖的降解;而Fe~(2+)和Mn~(2+)则起显著的促进作用.pH值、温度、金属离子等共同调控了endo-PG对果胶多糖的降解作用.     

Effect of hydroxyl radical on the scission of cellular wall polysaccharides in vitro of banana fruit at various ripening stages

Fruit ripening is generally attributed to disassembly of cellular wall, particularly due to solubilisation and depolymerisation of pectin and hemicellulose. Experiments were conducted to test effects of hydroxyl radicals (·OH) on the scission of cellular wall polysaccharides from pulp tissues of banana fruit at different ripening stage. Cellular wall materials were isolated from pulp tissues of banana fruit at different ripening stages. Two pectic fractions, water soluble pectin (WSP) and acid soluble pectin (ASP), and two hemicellulosic fractions, 1 M KOH soluble hemicellulose (HC1) and 4 M KOH soluble hemicellulos (HC2), were obtained from the cellular wall materials from pulp tissues, respectively. Effects of ·OH induced by the Fenton reaction on the scission of pectin and hemicellulose in vitro were investigated. As fruit ripening progressed, the sugar components of the WSP, HC1 and HC2 attacked by ·OH showed obvious molecular-mass downshifts. Thus, ·OH caused the disassembly of polysaccharides (WSP, ASP, HC1 and HC2) from cellular walls of pulp tissues of banana fruit, demonstrated by the reduced molecular mass distribution. Moreover, ·OH production in pulp tissues increased significantly as banana fruit ripened, which further help account for the role of ·OH in accelerated fruit ripening.     

Structural features of pectic polysaccharides from the skin of Opuntia ficus-indica prickly pear fruits

After removal of the mucilage with water at room temperature, pectic polysaccharides were solubilized from Opuntia ficus-indica fruit skin, by sequential extraction with water at 60 degrees C (WSP) and EDTA solution at 60 degrees C (CSP). Polysaccharides with neutral sugar content of 0.48 and 0.36 mol/mol galacturonic acid residue were obtained, respectively, in the WSP and CSP extracts. These pectic polysaccharides were de-esterified and fractionated by anion-exchange chromatography, yielding for each extract five fractions, which were thereafter purified by size-exclusion chromatography. Two of these purified fractions were characterized by sugar analysis combined with methylation and reduction-methylation analysis. The study was then supported by (1)H and (13)C NMR spectroscopy. The results showed that the water-soluble fraction WSP3 and the EDTA soluble fraction CSP3, consisted of a disaccharide repeating unit -->2)-alpha-l-Rhap-(1-->4)-alpha-d-GalpA-(1--> backbone, with side chains attached to O-4 of the rhamnosyl residues. The side chains contained highly branched alpha-(1-->5)-linked arabinan and short linear beta-(1-->4)-linked galactan.     

香蕉果实成熟软化过程中β-D-木聚糖苷酶活性变化

β-D-木聚糖苷酶是细胞壁半纤维素中阿拉伯木聚糖和木聚糖残基降解的主要酶,对香蕉贮藏过程中果皮、果肉中β-D-木聚糖苷酶活性以及果实硬度、呼吸强度和乙烯释放量的变化进行测定分析。结果显示:β-D-木聚糖苷酶活性在果实贮藏初期的变化很小,到果实硬度开始急剧下降时迅速增加,其增加量在果皮和果肉中分别为12和22倍以上,且果肉中的酶活性大于果皮中;乙烯吸收剂处理延缓了香蕉果实呼吸和乙烯的高峰出现以及果实硬度、果肉和果皮中β-D-木聚糖苷酶活性变化的速度和幅度,但并不改变其活性的变化趋势。结果证明,β-D-木聚糖苷酶能诱导香蕉果实成熟,在果实软化中起着十分重要的作用,且其活性受乙烯的调节。     

香蕉果实XET cDNA的克隆及其在成熟软化的果实果皮和果肉中的表达分析

木葡聚糖内糖基转移酶(Xyloglucan endotransglycosylase,XET)通过分解细胞壁半纤维素多糖的主要成分--木葡聚糖而参与果实软化.为了阐明香蕉(Musa acuminata.Colla cv.GrandNain)果实成熟过程中的软化与细胞壁代谢酶XET基因表达模式的关系,采用RT-PCR和RACE-PCR方法,首次从成熟香蕉果实果肉中分离了编码XT基因的全长cDNA(MA-XET1,全长1 095 bp).序列分析表明,MA-XET1的5'端和3'端的非翻译区分别为66 bp和1 89bp,该片段含有一个完整的开放读码框,编码280个氨基酸,推导的MA-XET1蛋白质中存在XET蛋白的催化活性部位DEIDFEFL.Southern杂交表明,MA-XET1在香蕉基因组中由多拷贝基因编码.Northern分析显示,跃变前期的果肉中,不能检测MA-XET1基因的表达,跃变期的果实果肉中MA-XET1表达增加,跃变后期该基因表达略有减弱;在跃变前期的果实果皮中,MA-XET1的积累较低,跃变期的果实果皮中积累大幅增加,而后迅速下降.Propylene(丙烯,乙烯的类似物)处理降低香蕉果实果皮和果肉的硬度,而且propylene促进MA-XET1在果皮和果肉中的积累.这些结果表明,MA-XET1参与香蕉果实成熟过程中的果皮和果肉软化,并且,MA-XET1的表达在转录水平上受乙烯调控.     

Endo-1,4-[beta]-Glucanase,Xyloglucanase, and Xyloglucan Endo-Transglycosylase Activities Versus Potential Substrates in Ripening Tomatoes

In ripening fruits of tomato (Lycopersicon esculentum L. var 83-G-38), the amounts of cellulose and xyloglucan (XG) remained constant during tissue softening, but the relative molecular weight (Mr) of XG decreased markedly and the Mr of cellulose declined slightly. These changes could have been due to activities of non-specific endo-1,4-[beta]-glucanases and/or buffer-soluble XG endo-transglycosylase, both of which increased when tissue firmness declined most rapidly. Tomato extracts also reduced the viscosity of XG solutions, especially in the presence of added XG oligosac-charides. This depolymerizing (XGase) capacity differed from [beta]-glucanase and XG transglycosylase activity (a) by being almost entirely buffer insoluble, and (b) by declining precipitously during fruit softening. Although it disappeared from ripe fruit, XGase may have functioned in promoting wall loosening at earlier stages of fruit development when its activity was highest. By contrast, during aging of fruit in the ripening-inhibited mutant rin there was no change in Mr of XG or cellulose, and activities of [beta]-glucanases and XG transglycosylase were lower than in wild-type tomato. Nevertheless, some softening of the fruit did take place over time and XG amounts declined, possibly because high XGase activity was maintained in the mutant, unlike in wild-type fruit.     

香蕉果实软化相关β-半乳糖苷酶基因的克隆及其表达分析

β-半乳糖苷酶（β-galactosidase）通过分解细胞壁半纤维素切除半乳糖键而参与果实软化。为了阐明香蕉（Musasp．）果实成熟过程中的软化与细胞壁代谢酶β-半乳糖苷酶基因表达之间的关系,采用RT-PCR方法,从成熟香蕉果实果肉中分离了编码β-半乳糖苷酶基因的部分cDNA（MA-Gal）,序列分析表明,MA-Gal包含927bp,编码309个氨基酸,包含5个β-半乳糖苷酶结构域（典型真核生物中β-半乳糖苷酶包含7个结构域）,推导的MA-Gal蛋白质中有β-半乳糖苷酶蛋白的催化活性部位GGPIILSQIENEY（F）;系统进化树分析结果表明MA-Gal属于第一类β-半乳糖苷酶基因（该类主要在果实中表达）;β-半乳糖苷酶活性和硬度的变化表明其与香蕉果实硬度变化密切相关;Northern分析显示,跃变前期的果肉中,MA-Gal基因的表达量很低,后随着果实的软化表达量不断增加,并在呼吸跃变后达到最高。所有结果表明,MA-Gal参与香蕉果实成熟过程中的软化。     

Changes in cell-wall polysaccharides from the mesocarp of grape berries during veraison

Softening of grape berries ( Vitis vinifera L. × V. labruscana cv. Kyoho) was evaluated by studying changes in composition and degradation of cell-wall polysaccharides. The grape berry softens at the beginning of the second growth cycle many weeks before harvest. The softening stage is called 'veraison' by viticulturists. On day 50 after full bloom, green hard berries (before veraison [BV]), softening berries (veraison [V]) and partly peel colored berries (C) were selected from the same clusters. In addition, mature berries (M) were collected on day 78 after full bloom. Mesocarp tissues at each stage were fractionated into hot water-soluble (WS), hot EDTA-soluble (pectin), alkali-soluble (hemicellulose) and residual (cellulose) fractions. Neutral and acidic sugar contents of WS and pectin fractions decreased only after the V stage, while the neutral sugar content of the hemicellulose fraction decreased from the BV to V stages. Cellulose content constantly decreased as the berry ripened, but the large decrease was found from the BV to V stages. Molecular masses of pectic and hemicellulosic polysaccharides decreased from the BV to V stages. Hemicellulosic xyloglucan was markedly depolymerized from the BV to V stages. The neutral and acidic sugar composition of each fraction changed little during the berry ripening. These data indicated that softening of berry during veraison involved the depolymerization of pectin and xyloglucan molecules and decrease in the amounts of hemicellulose and cellulose.     

Changes in the cell-wall polysaccharides of outer pericarp tissues of kiwifruit during development.

Changes in pectin, hemicelluloses and cellulose in the cell walls of outer pericarp tissues of kiwifruit (Actinidia deliciosa cv. Hayward) were determined during development. An extensive amylase digestion was employed to remove possible contaminating starch before and after fractionation of wall polysaccharides. An initial treatment of crude cell walls with alpha-amylase and iso-amylase or DMSO, was found to be insufficient removing the contaminating starch from wall polysaccharides. After EDTA and alkaline extraction, the pectic and hemicellulose fractions were again treated with the combination of alpha-amylase and iso-amylase. The amounts of predominant pectic sugars Gal, Rha and Ara, unaffected by the first and second amylase digestion, decreased markedly during the early fruit enlargement (8-12 weeks after anthesis, WAA), then increased during 16-20 WAA, and finally declined during fruit maturity (20-25 WAA). The molecular-mass of pectic polysaccharides decreased during fruit enlargement (8-16 WAA), and then changed little during fruit maturity. The higher molecular-mass components of hemicelluloses in HC-I and HC-II fractions detected at the early stage of fruit enlargement (8-12 WAA) were degraded at the late stage of fruit enlargement (16 WAA), but then remained stable at the much lower molecular-mass till fruit maturity. The amount of Xyl in the HC-II fraction decreased during the early fruit enlargement and fruit maturity, an observation that was consistent with xyloglucan (XG) content. The gel permeation profiles of XG showed a slight increase in higher molecular-mass components during 8-12 WAA, but thereafter there was no significant down-shift of molecular-mass until harvest time. The cellulose fraction increased steadily during fruit enlargement through maturity, but the XG contents in HC-I and HC-II fractions remained at a low level during these stages. Methylation analysis of HC-I and HC-II fractions confirmed the low level of XG in the hemicellulosic fractions. It was suggested that pectin in the outer pericarp of kiwifruit was degraded at the early stage of fruit enlargement, but XG remains constant during fruit enlargement and maturation.