单宁细胞形态与部分柿属种及品种相关性研究

采用软柿果肉直接压涂法,在光学显微镜下对204个柿属种及品种果实中的单宁细胞形态特征进行观察分析.结果显示:(1)在6个种柿属植物果实中,均含有单宁细胞,其外形大多属于短形和近圆形,但在数量、大小和颜色上存在差异,其中,油柿(Diospyros oleifera Cheng.)、君迁子(D.lotus Linn.)、柿(D.kaki Thunb.)、浙江柿(D.glaucifolia Metc.)的单宁细胞通常无色,而黑柿(D.nitida Mcrr.)为黄绿色,乌材(D.eriantha Champ.)为紫红色,乌柿(D.eathayensis Stheward.)为淡紫色;单宁细胞从大到小依次为油柿>君迁子>浙江柿>乌材>黑柿>乌柿.(2)单宁细胞在不同品种类型间差异明显,其中涩柿单宁细胞多为无色,单宁细胞比较宽大;甜柿品种均会出现褐变的单宁细胞,单宁细胞较小或瘦长;完全甜柿品种大多存在着凝固型褐变单宁细胞,仅少数凝聚呈球形,且单宁细胞分散存在于果肉中,果肉中的褐斑较细小;不完全甜柿在种子周围的褐斑处,可以看到大量的表面凹形且褐变的收缩型单宁细胞,且常以单宁细胞束的形态存在于果肉中,使果肉中的褐斑大而密;原产我国的完全甜柿中不存在凝聚型的单宁细胞,只有凝固型的单宁细胞.(3)聚类分析结果表明,单宁细胞的特征可以作为不同类型柿属种的分类依据.     

热处理对脱涩后火柿生理生化变化的影响

以CO2脱涩后的火柿为试材,研究了脱涩后火柿在5℃条件下贮藏过程中主要生理生化变化及35℃热处理对其软化生理的影响。结果表明,柿果可溶性单宁含量的继续下降是柿果软化的一个原因;乙烯在脱涩柿果的软化中起着主导作用,呼吸速率及PE、PG和纤维素酶活性的变化与乙烯存在着紧密的关系。35℃热处理抑制了脱涩火柿可溶性单宁含量的下降速率,使乙烯生成量降低和乙烯峰出现时间推迟,刺激了早期的呼吸速率,抑制了后期的呼吸速率,降低了PE、PG和纤维素酶的活性,从而达到了延缓脱涩柿果轮化的目的。     

CO2和1-MCP组合处理对磨盘柿贮藏效果的影响

以磨盘柿采后果实为材料,研究1-MCP的处理以及CO2脱涩与1-MCP组合处理对磨盘柿室温和0~1℃贮藏过程中果实硬度、可溶性单宁含量、总抗坏血酸(TAA)含量以及抗氧化活性的影响。结果显示:(1)贮藏过程中柿果实硬度随时间延长呈下降趋势,室温贮藏CO2处理5 d、CO2处理后进行1-MCP(CO2//1-MCP)处理10 d、1-MCP处理后进行CO2(1-MCP//CO2)处理15 d、CO2与1-MCP同时(CO2+1-MCP)处理30 d均完全软化,0~1℃贮藏75 d后硬度最高的是CO2+1-MCP处理(12.43 kg.cm-2),最低的是CO2//1-MCP处理(2.80 kg.cm-2),甚至低于CO2处理(5.71 kg.cm-2);(2)柿果实可溶性单宁和总抗坏血酸(TAA)含量与脱涩有关,CO2处理、CO2//1-MCP处理、1-MCP//CO2处理和CO2+1-MCP处理大幅低于CK和1-MCP处理,1-MCP处理抑制了可溶性单宁和TAA含量的下降;(3)室温贮藏中除CO2处理之外的处理柿果实总酚含量呈小幅上升趋势,0~1℃贮藏中CK、1-MCP处理、1-MCP//CO2处理和CO2+1-MCP处理的总酚含量稳定在9.91~12.38 mg.g-1FW之间,CO2处理和CO2//1-MCP处理于30 d之后迅速下降,至75 d时分别只有6.83和6.32 mg.g-1FW;(4)各组处理柿果实ABTS自由基清除能力和氧自由基清除能力值(ORAC)的变化趋势与总酚含量大致相当。研究发现,1-MCP能有效阻止磨盘柿果实贮藏期间的硬度、可溶性单宁含量、TAA含量以及抗氧化活性的下降,CO2脱涩与1-MCP处理的不同顺序对硬度和抗氧化活性影响巨大,但对可溶性单宁和TAA含量影响甚微;先CO2脱涩后1-MCP处理对磨盘柿贮藏效应影响不大,1-MCP处理和高浓度CO2脱涩同时进行是磨盘柿脱涩保鲜的最优方案。     

‘恭城月柿''不同生育期果实和叶片单宁组分含量的变化

柿单宁是一种重要的食品功能成分,广泛应用于食品、医疗等领域,研究柿单宁的生物生产特性对其产业应用具有重要意义。该研究以广西特色涩柿品种‘恭城月柿’为材料,测定了幼果期、膨大期、成熟前期、成熟期和成熟后期柿果肉、果皮和叶片的可溶性单宁含量,并利用液质联用色谱对全部样品中6种单宁组分[没食子酸(GA)、没食子儿茶素(GC)、儿茶素(CA)、芦丁(Rutin)、表没食子儿茶素(EGC)和表儿茶素(EC)]进行了定量分析。结果表明:柿果肉和果皮中含有丰富的可溶性单宁,在膨大期含量最高,分别达到21.93 mg·g~(-1)和18.41 mg·g~(-1),随着果实进入成熟阶段显著下降;柿叶片的可溶性单宁含量较低,并且在整个生育期内无明显变化。另外,在柿果肉中检测到GA、GC和CA 3种单宁组分,其中GA含量显著高于其他组分;在柿果皮中检测到GA、GC、CA、Rutin和EGC 5种组分,其中CA含量最高,EGC仅在幼果期和膨大期有痕量检出;在柿叶中检测到全部6种单宁组分,其中整个生育期CA含量极高,EGC和EC均为痕量检出。由此可知,幼果期至膨大期的柿果实可产出较多可溶性单宁,其中果肉中含有较多的GA组分,果皮中含有大量的CA组分,柿叶片在整个生育期均可产出极为丰富的CA单宁组分。该研究为柿单宁的功能性开发利用提供基础数据。     

山茱萸果实发育过程中单宁物质的分布与积累特征

选取不同发育时期的山茱萸果实作为研究对象,采用果实形态观察法、显微及超微技术、组织化学定位法以及紫外分光光度计法对山茱萸果实发育过程中单宁物质分布及积累特征进行观察分析,并以单因素ANOVA检验不同发育时期单宁含量的差异,以揭示单宁物质在山茱萸果实发育中的变化规律,为山茱萸果实涩味调控机制研究提供理论依据。结果表明:(1)山茱萸果实发育过程中果皮颜色和果实体积变化明显,可将其发育过程划分为幼果期、中果期、成熟期3个时期;单宁物质主要分布在山茱萸果实中果皮的单宁细胞中。(2)在山茱萸果实发育过程中单宁细胞数目呈先增后减的变化趋势,幼果期单宁细胞从无到有,随着果实发育单宁细胞数目不断增多,至中果期单宁细胞数目开始减少。(3)单宁含量的变化规律与单宁细胞数目的变化一致,单宁含量在花后120 d时达到最多,随后逐渐减少。(4)单宁物质首先在细胞质的小液泡中积累,中央大液泡形成后则为单宁物质积累的主要场所,其积累形态主要有颗粒状、不规则状和板块状3种;单宁细胞中线粒体数目较多,中果期后期及成熟期在中央大液泡液泡膜附近有电子致密物质积累。研究认为,山茱萸果实中中果皮薄壁细胞为单宁物质积累的专属细胞,即单宁细胞,单宁物质的合成运输与液泡、囊泡以及线粒体的作用密切相关;成熟期山茱萸果实总单宁含量降低,涩味降低,表明单宁物质积累的动态变化与植物对环境的适应性和果实涩味息息相关,可结合代谢组和转录组的方法对山茱萸果实中单宁物质的合成机制进行进一步研究。     

杉木种子涩籽地理流行趋势的研究

本文应用趋势面分析福建省杉木种子涩籽地理流行趋势,建立涩籽地理流行数学模型,阐述其流行规律,绘制山涩籽地理流行图,并将其划分成四个区：1．闽西南杉木涩籽流行递减区;2．闽东南杉木涩籽流行递增区;3．闽北杉木涩籽流行递减区：4,闽中北闽西北杉木涩籽流行渐稳区。同时建议将大型杉木种子园建在建瓯县。     

柿子脱涩及保鲜方法

在真空条件下,人们可用多种不同的方法去掉柿子中的涩味。其中,最常用的方法是造成富含二氧化碳的大气环境(>80％CO_2),并持续1～2天。无论用自然方法还是用人工方法脱涩,都必须经过在真空条件下柿内形成乙醛的过程。乙醛与单宁反应形成不溶性胶,带有涩味的水果中所含的可溶性单宁与乙醛发生聚合反应形成无涩味的不溶性化合物。因此,脱涩的速度和程度取决于在二氧化碳或氮气里(或两者的混合物),柿子内部乙醛形成的水平。     

杉木败育种子及其涩粒物质的研究

杉木种子的涩粒(败育种子)约占种子数的30—70%,涩粒率高是杉木种子发芽率低的根本原因之一。试验表明,立地条件、授粉方式、胚胎发育、光照、涩粒物质及母株的遗传性等都与涩粒的形成有着密切关系。涩粒物质的成分很复杂,其中含萜类物质。还含有多种矿质元素。通过采取多种综合措施,可以有效地提高种子的质量,较大幅度地降低涩粒率。     

杉木种子涩籽地理分布的空间插值法研究

应用距离反比插值法进行杉木种子涩籽率的空间插值,并提出一种改进的距离反比插值法(MID),其包含了距离反比法(ID)和距离平方反比法(IDS)。根据福建省25个杉木种子产地的涩籽率观察值,建立了杉木种子涩籽率的改进距离反比插值模型。改进距离反比插值法经交叉验证,其平均误差及平均误差平方的平方根均较距离反比法和距离平方反比法更小,具有较高的精度,平均精度达88．91％,可应用于杉木种子涩籽的空间内插。这不仅为杉木种子涩籽的空间插值提供了一种新方法,而且为地理信息系统(Geographic information system,GIS)辅助的杉木种子涩籽区域分布与流行趋势研究提供了理论依据,为福建省杉木种子园的合理布局与优化奠定基础。     

小型猪涩脉模型的建立与评价

目的基于小型猪的生理特点及循环系统与人类极为相似的特征，通过药物对其造模，建立一种新型的脉诊动物教学模型并对其进行评价。方法选用中华实验用成年小型猪6只（体重20±1．6kg，选取心率〈100次/min）。采用戊巴比妥钠麻醉，定位取脉位置，切脉后，用NX-8型脉诊仪同步采取正常脉图、二导心电图，血流动力学参数，血流变学指标。耳大静脉心得安注射液及葡聚糖T-500静脉给药进行造模，观察造模后一般情况（心率、皮肤颜色）及指下脉形变化，同步描记脉图及二导心电图，涩变后记录数值：不同时间点的心率变化；主波高（HB）、升支斜率（Sup）、降支斜率（Sdown）、降中峡位置（即降中峡高，主波高HE/HB）、升支时间，脉波周期（TAB/TAG）、主波峰时间，脉波周期（TB/TAG）；并采集造模后血流动力学及血流变学各项参数。结果6头小型猪均造模成功，指下感觉涩变明显；脉图变化显著；与造模前相比HB、Sup、Sdown、HE/HB、TAB/TAG、TB/TAG差异均有显著性（除SdownP〈0．05外，其余均P〈0．01）。血流动力学及血流变学相关参数差异也均有显著。讨论通过运用小型猪建立涩脉模型成功、方法可靠，小型猪应用于脉诊教学实验方法简便、损伤小、可重复运用，有充分的可行性，以此为基础可以建立多种脉诊模型，以补充脉诊实验教学中的空白。     

Effects of persimmon tannin on survival and reproduction of Halyomorpha halys (Hemiptera: Pentatomidae)

Halyomorpha halys (Hemiptera: Pentatomidae) occurs in sweet persimmon orchards from late July to early September and its damage level is negligible before mid‐July. It rarely appears in persimmon orchards after mid‐September, but is easily found in soybean fields. From the phenological observation of H. halys, it is assumed that seasonal migration of H. halys to sweet persimmon orchards may be related to the content of soluble tannin in sweet persimmon fruits. The feeding preference of H. halys was evaluated on sweet and astringent persimmon fruits, and the effects of persimmon fruits and purified persimmon tannin were tested on the survival of H. halys. Numbers of visiting bugs and feeding spots on the fruits were significantly higher on sweet persimmon than those on astringent persimmon. There was no significant difference in survivorships of H. halys fed sweet persimmon and soybean. However, the number of eggs laid by sweet persimmon fed adult females was significantly lower than those fed soybean. The higher concentration of persimmon tannin caused higher mortality of H. halys. Male and female H. halys fed with 3% persimmon tannin showed the lowest survival. Longevities of male H. halys fed with 0, 0.01, 0.1 and 1% persimmon tannin were 14.9, 12.7, 14.5 and 16.3 days, respectively. However, males fed with 3% tannin lived only 6.4 days. The longevities of female H. halys were similar to those of male (10.1, 14.4, 13.8, 12.0 and 6.1 days for 0, 0.01, 0.1, 1 and 3% tannin concentration, respectively). These results suggest that H. halys invades sweet persimmon orchards during late July and early September when more suitable foods, e.g. soybean, are in short supply and sweet persimmon fruits become suitable for feeding (e.g. lower tannin content).     

Carbon dioxide assimilation during postharvest removal of astringency from persimmon fruit

Application of anaerobic conditions with CO₂ or N₂ atmospheres to remove astringency from harvested persimmon fruit ( Diospryros kaki L. cv. Triumph), caused production of more acetaldehyde under CO₂ than under N₂, ¹⁴CO₂ applied in a 100% CO₂ atmosphere, for 48 h to astringent persimmon fruits was incorporated mainly into malate and very little into other metabolites, such as carbohydrate or amino acids. Application of malate or pyruvate to pulp discs of astringent persimmons caused an immediate rise in acetaldehyde production. The higher levels of acetaldehyde produced by whole fruits held in a CO₂ atmosphere, than by fruits held in a N₂ atmosphere, can be explained through fixation of atmospheric CO₂ into malate, leading to acetaldehyde production.     

Volatile compounds associated to the loss of astringency in persimmon fruit revealed by untargeted GC–MS analysis

High resolution volatile profiling (67 compounds identified) of fruits from 12 persimmon cultivars was established and used to characterize the different astringency types of persimmon fruit before and after deastringency treatment. Analysis of the volatile profile of fruit enables us to differentiate between cultivars that at the moment of harvest produced non-astringent fruit (Pollination Constant Non Astringent—PCNA-type) from astringent ones (non-PCNA-type). Fruit failing to accumulate astringent compounds naturally (PCNA fruit) showed high levels of 3(2H)-benzofuranone, while this compound was not detected in any astringent type fruit (non-PCNA). In addition to this, PCNA cultivars also showed at harvest higher accumulation of benzeneacetaldehyde and lipid-derived aldehydes (hexanal, heptanal, octanal and decanal) than non-PCNA fruit. The application of postharvest deastringency treatment to all non-PCNA cultivars resulted on an important insolubilization of tannins. In general the CO₂-treatment enhanced the levels of acetaldehyde, however those cultivars showing high levels of dihydrobenzofuran at harvest did not present an increment of acetaldehyde. In contrast, all non-PCNA cultivars exhibited an important accumulation of lipid-derived aldehydes due to CO₂-treatment. Therefore, we propose that lipid-derived aldehydes (mainly decanal, octanal and heptanal) may be playing a role in the astringency loss. Our results suggest that 3(2H)-benzofuranone, benzeneacetaldehyde and lipid-derived aldehydes could be used as markers for both natural and artificial loss of astringency.     

Turbidity as a measure of salivary protein reactions with astringent substances

Binding of tannins to proline-rich proteins has been proposed as an initial step in the development of astringent sensations. In beer and fruit juices, formation of tannin-protein complexes leads to the well-known effect of haze development or turbidity. Two experiments examined the development of turbidity in human saliva when mixed with tannins as a potential in vitro correlate of astringent sensations. In the first study, haze was measured in filtered human saliva mixed with a range of tannic acid concentrations known to produce supra-threshold psychophysical responses. The second study examined relationships among individual differences in haze development and the magnitude of astringency ratings. Mostly negative correlations were found, consistent with the notion that high levels of salivary proteins protect oral tissues from the drying effects of tannic acid.     

Isolation and expression of gene encoding leucoanthocyanidin reductase from <Emphasis Type="Italic">Diospyros kaki</Emphasis> during fruit development

Leucoanthocyanidin reductase (LAR) converts leucoanthocyanidin to (+)-catechin, a precursor of proanthocyanidins abundant in Japanese persimmon (Diospyros kaki Thunb.) fruits. A putative LAR gene (DkLAR) was isolated by rapid amplification of cDNA ends from young fruits. The full-length cDNA of DkLAR gene was 1 356 bp long and encoded an open reading frame of 349 residues. The deduced DkLAR protein was closely related to the homolog in other plant species. The expression of the DkLAR gene in Chinese pollination-constant non-astringent (PCNA) genotype was coincident with the tannin cell development, but was not in Japanese PCNA and Chinese pollination-variant astringent (PCA) genotypes.     

SELECTION OF AN ASTRINGENCY REFERENCE STANDARD FOR THE SENSORY EVALUATION OF BLACK TEA

Astringent and bitter sensations are characteristic sensory qualities of black tea. Three different classes of potential astringent reference standards (two concentrations each of alum and tannic acid and three fruit juices) were evaluated in this study. The perceived astringency, bitterness and sourness of each were profiled using computerized time-intensity and compared with the astringent intensity of a standardized brew of black tea. The differences in temporal profiles of potential reference standards across taste attributes were evident and intensity ratings were found to be dependent upon the stimulus and its concentration. Both concentrations of tannic acid were evaluated as the highest in perceived bitterness. For the juices, a strong sour taste was perceived in addition to astringency. It was concluded that the best reference standard for the astringency of black tea is a solution of 0.7 g/L alum as it is low in perceived bitterness and sourness.     

Expression of ethylene response genes during persimmon fruit astringency removal

Thirteen ethylene signaling related genes were isolated and studied during ripening of non-astringent ‘Yangfeng’ and astringent ‘Mopan’ persimmon fruit. Some of these genes were characterized as ethylene responsive. Treatments, including ethylene and CO₂, had different effects on persimmon ripening, but overlapping roles in astringency removal, such as increasing the reduction in levels of soluble tannins. DkERS1, DkETR2, and DkERF8, may participate in persimmon fruit ripening and softening. The expression patterns of DkETR2, DkERF4, and DkERF5 had significant correlations with decreases in soluble tannins in ‘Mopan’ persimmon fruit, suggesting that these genes might be key components in persimmon fruit astringency removal and be the linkage between different treatments, while DkERF1 and DkERF6 may be specifically involved in CO₂ induced astringency removal. The possible roles of ethylene signaling genes in persimmon fruit astringency removal are discussed.     

基于网络药理学的柿果药理功能定位及作用机制

基于网络药理学,通过国内外文献检索获取柿果中的化合物,采用Swiss target prediction数据库对化合物进行潜在靶点垂钓以探讨柿果的药理功能定位及作用机制。以Cytoscape软件构建化合物-靶点网络,靶点-疾病名称-疾病分类网络,同时对靶点进行蛋白相互作用(PPI)网络构建,采用DAVID数据库对靶点进行通路富集分析。本研究共收集到柿果中16个化合物,可作用于68个靶点,这些靶点主要作用于心血管疾病、神经精神性疾病等。PPI网络图包含84个节点,226条边,其中degree值排前10的关键蛋白分别为ERS1、PGS2、MMP2、TIMP1、MMP9、MMP1、AR、SLC6A3、PRKCB、CYP19A1。上述靶点可调节氮素代谢、血清素能突触以及TRP通道炎症介质的调节等信号通路。本研究为阐明柿果的药理功能定位及其作用机制研究提供了可靠的依据。     

Xyloglucan-derived oligosaccharides induce ethylene synthesis in persimmon (Diospyros khaki L.) fruit

In this work the effect of injection of xyloglucan-derived oligosaccharides (XGOs) into whole persimmon (Diospyros khaki L.) fruits on ethylene biosynthesis was investigated. Fruits collected during different ripening stages produced low levels of ethylene without a climacteric-like peak. Pretreatment of these fruits with 10 cm³ C2H4 m^-3 for 8 h stimulated little or no endogenous ethylene production. However, when persimmon fruits were injected with a mixture of XGOs a burst in ethylene production was observed compared with water-injected control fruits or fruits injected with different monosaccharide solutions. In order to study the influence of oligosaccharide structure and fruit ripening stage on the ability of XGOs to induce ethylene synthesis, fucosylated and non-fucosylated XGOs were injected into persimmon fruits harvested at two different ripening stages. Both oligosaccharide structures were able to induce ethylene production. Induction of ethylene by XGOs was much more evident in fruits harvested later in time, indicating that the process is developmentally regulated. The levels of 1-aminocyclopropane-1-carboxylic acid (ACC) in injected persimmon fruits were also examined. This study showed that the increase in the rate of ethylene biosynthesis induced by XGOs was accompanied by the accumulation of its metabolic precursor ACC.