利用隶属函数值法对玉米成熟期抗旱性的综合评价

以10个不同基因型玉米为试验材料,在宁夏中部干旱带,通过测定不同品种成熟期的K~+/Na~+、光合气体交换参数、农艺性状及产量性状等指标进行抗旱性鉴定,并利用主成分分析和隶属函数法对其抗旱性强弱进行综合评价。结果表明,干旱胁迫地和对照田中,各参试品种的光合特征参数、K~+/Na~+及形态性状等均有明显差异,且干旱胁迫条件下的净光合速率(P_n)、株高、产量均显著低于对照(P0.05)。同一品种不同器官的K~+/Na~+值表现为茎基茎叶根,表明植株体内的离子吸收和运输具有选择性,茎基具有贮存K~+的作用,根具有贮存Na~+的功能。不同基因型玉米品种的抗旱能力根据综合评价值(D值)大小,综合评价其抗旱性强弱顺序为A111-3/H242HoViY1北21/A58A3/A18高WY2宁3/1522H237/A18桂青贮1号H237/1506A3/A18。     

盐胁迫下钾吸收途径的抑制对小麦叶片K~+、Na~+含量的影响

为研究盐胁迫下小麦维持钾、钠平衡的生理机制,以耐盐小麦沧麦6005和盐敏感小麦矮抗58为材料,利用TEA、NEM、Ba(NO_3)_2三种药物分别抑制钾离子通道、钾载体及非选择性阳离子通道,测定正常及盐胁迫下小麦叶片K~+、Na~+含量,比较耐盐性不同的小麦品种在K~+、Na~+吸收中的差异。结果显示,盐胁迫下,沧麦6005和矮抗58叶片K~+含量下降,Na~+含量增加;沧麦6005叶片Na~+含量低于矮抗58,K~+/Na~+比值高于矮抗58。正常条件下,NEM、TEA处理均可降低沧麦6005和矮抗58叶片K~+含量,NEM处理较TEA处理效果更为明显;TEA处理显著降低了盐胁迫下矮抗58叶片K~+含量,而NEM处理则明显降低了盐胁迫下沧麦6005的叶片K~+含量;TEA、NEM、Ba(NO_3)_2处理降低了盐胁迫下矮抗58叶片Na~+含量,仅NEM处理降低了沧麦6005叶片Na~+含量。综上所述,正常条件下,钾载体是小麦K~+吸收的主要方式;盐胁迫下,耐盐品种和盐敏感品种K~+吸收途径不同,耐盐品种的NSCCs和钾离子通道具有更强的"拒钠"能力。     

Differential Sensitivity of Rice Cultivars to Salinity and Its Relation to Ion Accumulation and Root Tip Structure

Abstract

Effects of NaCl on the growth, ion content, root cap structure and Casparian band development were examined in four rice (Oryza sativa L.) cultivars with different salt resistance (salt-sensitive indica-type IR 24 and japonica-type Nipponbare and salt-resistant indica-type Nona Bokra and Pokkali). Experiments were conducted to find the differences in salinity resistance during early seedling and developed seedling stages among the cultivars. For salinity treatment, sodium chloride (NaCl) was added to nutrient solution at concentrations of 0, 25 and 50 mM for 7 days from germination to the 7th day (early seedling stage) or from the 7th day to 14th day (developed seedling stage). Growth inhibition by salinity was more prominent in the early seedling stage than in the developed seedling stage. Based on the growth, the order of the sensitivity was IR24 > Nipponbare > Nona Bokra > Pokkali. The growth of NaCl-treated rice cultivars relative to control was significantly and negatively correlated with the Na⁺ content and Na⁺/K⁺ ratio in roots and shoots in both stages. Scanning electron microscopic observation revealed that the root cap tissues proliferated and extended to the basal part of the root tip by salinity. The length of root cap was, however, reduced by 50 mM NaCl in sensitive cultivars due to peeling off. An endodermal Casparian band was formed in the basal region of the root tip. Development of the Casparian band was more prominent in sensitive cultivars than in tolerant cultivars. Root cap proliferation might be related to NaCl resistance in rice seedlings, but the Casparian band may not function efficiently in Na⁺ exclusion. Essentially the present results suggest that exclusion of Na⁺ from roots plays a critical role in expression of Na⁺ resistance in rice seedlings and the root cap is important for Na⁺ exclusion.     

Comparative effects of salt and alkali stresses on organic acid accumulation and ionic balance of seabuckthorn (Hippophae rhamnoides L.)

Seabuckthorn (Hippophae rhamnoides L.) is uniquely capable of growing well under various extreme environmental conditions, such as water deficit, salt stress, low temperature, and high altitude. It is of economic value and its berries are used in cosmetics and pharmaceutical products. In this study, we compared the effects of salt stresses (9:1 molar ratio of NaCl to Na₂SO₄, pH 6.48–6.65) and alkali stresses (9:1 molar ratio of NaHCO₃ to Na₂CO₃, pH 8.70–8.88) on the levels of inorganic ions and organic acids in H. rhamnoides L. to elucidate the physiological mechanism by which it tolerates salt or alkali stress (high pH). The results showed that, in leaves and stems under alkali stress, the Na⁺ content increased to a much greater extent than under salt stress. Neither salt nor alkali stress decreased the K⁺ content in leaves and stems; however, in roots, the K⁺ content decreased sharply with increasing alkali stress, whereas it remained relatively unchanged with increasing salt stress. This revealed a specific mechanism of absorption or transport for Na⁺ and K⁺ that was affected strongly by alkali stress. The results indicated that accumulation of organic acid (OA) was a central adaptive mechanism by which H. rhamnoides maintained intracellular ionic balance under alkali stress. OA may play different roles in different organs during adaptation to alkali stress, and its percentage contribution to total negative charge was higher in leaf than in stem. H. rhamnoides accumulated mainly malate, oxalate, and citrate in leaves and stems; however, in roots, less malate and citrate was accumulated, and acetate accumulation was enhanced significantly, which indicated that roots and shoots use different mechanisms to modulate OA metabolism.     

Na+/K+-ATPase Activation by cAMP in the Midgut of Lutzomyia longipalpis (Lutz & Neiva, 1912; Diptera: Psychodidae)

Lutzomyia longipalpis (Lutz & Neiva, 1912) females have been intensively studied regarding the regulation of midgut pH. The mechanisms involved in pH regulation are complex, and some aspects remain to be clarified. Here, we investigated the role of the Na⁺/K⁺-ATPase pump as an electrochemical potential generator and its modulation by the second messenger cAMP in the midgut of female L. longipalpis. Our results suggest that not only may Na⁺/K⁺-ATPase be the main generator of an electrochemical potential across membranes in the midgut of female L. longipalpis, but also its activity is positively regulated by cAMP. cAMP-mediated Na⁺/K⁺-ATPase pump activity might be necessary to maintain the transport of the nutrients produced during blood digestion.     

Anti-hyperglycemic Effect of the Aqueous Extract of Banana Infructescence Stalks in Streptozotocin-induced Diabetic Rats

Water extract of banana (Musa sapientum) infructescence stalks has been used in folk medicine in the treatment of diabetes mellitus. This work aims at verifying the claimed effect and elucidating its possible mode of action. The extract was given in replacement of drinking water to diabetic rats, and its mechanism of action was studied by investigating its involvement in glucose transport in Caco-2 monolayers, and in rat jejuna using an in situ perfusion technique. Its effect on the Na⁺/K⁺ ATPase was studied by measuring the amount of inorganic phosphate liberated. The extract reduced significantly blood glucose levels in diabetic rats and glucose transport across rat jejuna and Caco-2 monolayers, and induced a 50 % decrease in their Na⁺/K⁺ ATPase activity. The extract did not induce any further decrease in jejunal glucose uptake in the simultaneous presence of phloridzin and phloretin, respective inhibitors of SGLT1 and GLUT2 transporters nor did it induce a change in the protein expression of SGLT1 and GLUT2. It was concluded that the extract acts by reducing the Na⁺/K⁺ ATPase activity of enterocytes and consequently the sodium gradient required for sugar transport by SGLT1, which leads to down-regulation of GLUT2 and contributes to the observed anti-hyperglycemic effect.     

Marine Toxins Targeting Ion Channels

This introductory minireview points out the importance of ion channels for cell communication. The basic concepts on the structure and function of ion channels triggered by membrane voltage changes, the so-called voltage-gated ion channels (VGICs), as well as those activated by neurotransmitters, the so-called ligand-gated ion channel (LGICs), are introduced. Among the most important VGIC superfamiles, we can name the voltage-gated Na⁺ (Na_V), Ca²⁺ (Ca_V), and K⁺ (K_V) channels. Among the most important LGIC super families, we can include the Cys-loop or nicotinicoid, the glutamate-activated (GluR), and the ATP-activated (P2X_nR) receptor superfamilies. Ion channels are transmembrane proteins that allow the passage of different ions in a specific or unspecific manner. For instance, the activation of Na_V, Ca_V, or K_V channels opens a pore that is specific for Na⁺, Ca²⁺, or K⁺, respectively. On the other hand, the activation of certain LGICs such as nicotinic acetylcholine receptors, GluRs, and P2X_nRs allows the passage of cations (e.g., Na⁺, K⁺, and/or Ca²⁺), whereas the activation of other LGICs such as type A γ-butyric acid and glycine receptors allows the passage of anions (e.g., Cl⁻ and/or HCO₃⁻). In this regard, the activation of Na_V and Ca_V as well as ligand-gated cation channels produce membrane depolarization, which finally leads to stimulatory effects in the cell, whereas the activation of K_V as well as ligand-gated anion channels induce membrane hyperpolarization that finally leads to inhibitory effects in the cell. The importance of these ion channel superfamilies is emphasized by considering their physiological functions throughout the body as well as their pathophysiological implicance in several neuronal diseases. In this regard, natural molecules, and especially marine toxins, can be potentially used as modulators (e.g., inhibitors or prolongers) of ion channel functions to treat or to alleviate a specific ion channel-linked disease (e.g., channelopaties).     

Vertical Distribution of Sodium in Roots of Rice Plants Exposed to Salinity as Analyzed by Cryo Time-of-Flight Secondary Ion Mass Spectrometry

Abstract

Distribution of Na⁺ along the root axis under salinity stress was analyzed in two rice (Oryza sativa L.) cultivars with different salt resistance (salt-sensitive IR 24 and salt-resistant Pokkali). Rice plants were grown hydroponically and NaCl was applied with nutrient solution at concentrations of 0, 25 and 50 mM for 7 d after germination. The distribution of Na⁺ in roots under salinity was analyzed by the cryo time-of-flight secondary ion mass spectrometry (cryo TOF-SIMS). The Na⁺ content in the root was higher in salt-sensitive IR 24 than in salt-resistant Pokkali under NaCl stress. The content was highest at the root tip and was decreased basipetally along the root axis. The difference in Na⁺ content between the cultivars was apparent in all regions from the root tip.     

Differences in the Vegetative Growth between Common and Tartary Buckwheat in Saline Hydroponic Culture

Abstract

Common buckwheat (Fagopyrum esculentum Moench cv. Tsushima) and Tartary buckwheat (F. tataricum (L.) Gaertn. cv. Pontivy) were grown in a nutrient solution with or without added NaCl to investigate interspecific differences in their responses to salinity, based on their dry-matter production. The mechanism of salt tolerance was also studied. Addition of 100 mM NaCl to the culture solution (salt treatment) lowered the plant growth rate to 48% and 16% of the control in Tsushima and Pontivy, respectively, and decreased the net assimilation rate and mean leaf area of Pontivy more severely than in Tsushima. The salt treatment decreased the leaf growth rate and leaf area per leaf to 30% and 72% of the control, respectively, in Tsushima, and to 12% and 52%, respectively, in Pontivy. It decreased the photosynthetic rate to 67% and 35% of the control, and stomatal conductance to 25% and 15% of the control in Tsushima and Pontivy, respectively. It also decreased the transpiration rate to 41% and 30% of the control in Tsushima and Pontivy, respectively, and increased the wateruse efficiency 1.6 times in Tsushima, but did not influence the wateruse efficiency in Pontivy. In the saline solution, the accumulation of Na⁺ in leaves and stem was greater in Pontivy than in Tsushima, but that in the roots, was greater in Tsushima than in Pontivy. In both species, Na⁺ accumulated rapidly in the leaves after removal of the roots in the saline solution. We conclude that the difference in salt tolerance between common and Tartary buckwheat may result from the difference in accumulation of Na⁺ in leaves and absorption of Na⁺ by the roots.     

菌肥与腐熟秸秆对盐碱地燕麦渗透生理特性及产量的影响

为明确菌肥和腐熟秸秆对盐碱地燕麦渗透生理及产量的调节作用,以燕麦品种白燕2号和草莜1号为材料,分析了菌肥(F)、腐熟秸秆(S)及二者配施(FS)对燕麦不同生育时期植株K⁺、Na⁺、可溶性糖、有机酸含量及株高和产量的影响。结果表明,苗期和拔节期燕麦各器官K⁺和Na⁺积累量较高,且茎和叶的K⁺、Na⁺积累量均明显高于根部。与空白对照(CK)相比,S和FS处理均显著提高各时期燕麦根、茎、叶K⁺含量,显著降低Na⁺含量。F、S和FS处理对各时期两个燕麦品种植株可溶性糖含量也有显著影响。F处理显著增加了抽穗期和灌浆期两个品种植株总有机酸含量,尤其是草酸、乙酸、柠檬酸比例提高;S和FS处理下抽穗期燕麦植株总有机酸含量在第一年较CK显著提高,而第二年显著降低,主要是由草酸变化引起。F、S及FS处理在第二年均显著提高两个品种的籽粒、鲜草和干草产量,其中FS处理的增产效果最好。这说明腐熟秸秆配施菌肥有助于增强盐碱地燕麦植株的渗透调...     

Effect of Green Tea and Vitamin E Combination in Isoproterenol Induced Myocardial Infarction in Rats

The present study was aimed to investigate the combined effects of green tea and vitamin E on heart weight, body weight, serum marker enzymes, lipid peroxidation, endogenous antioxidants and membrane bound ATPases in isoproterenol (ISO)-induced myocardial infarction in rats. Adult male albino rats, treated with ISO (200 mg/kg, s.c.) for 2 days at an interval of 24 h caused a significant (P < 0.05) elevation of heart weight, serum marker enzymes, lipid peroxidation and Ca⁺² ATPase level whereas there was a significant (P < 0.05) decrease in body weight, endogenous antioxidants, Na⁺/ K⁺ ATPase and Mg⁺² ATPase levels. Administration of green tea (100 mg/kg/day, p.o.) and vitamin E (100 mg/kg/day, p.o.) together for 30 consecutive days and challenged with ISO on the day 29th and 30th, showed a significant (P < 0.05) decrease in heart weight, serum marker enzymes, lipid peroxidation, Ca⁺² ATPase and a significant increase in the body weight, endogenous antioxidants, Na⁺/K⁺ ATPase and Mg⁺² ATPase when compared with ISO treated group and green tea or vitamin E alone treated groups. These findings indicate the synergistic protective effect of green tea and vitamin E during ISO induced myocardial infarction in rats.     

Salt tolerance variability amongst Argentine Andean potatoes (Solanum tuberosum L. subsp.andigena)

Summary Salt tolerance in Andean potatoes can be a source for this trait in cultivated potatoes or be exploited to expand potato cultivation to more marginal areas. The purpose of this study was to determine salt tolerance in twelve Argentine Andean potato varieties through the response of nodal segments cultured in vitro and to evaluate whether changes in Na⁺, K⁺ or proline accumulation associate with salt tolerance. Genotypes Bianca, Airampia, Sisa Sani and Cuarentilla were the most tolerant. The most susceptible varieties were Balcacha, Runa Rosada, Collareja, Overa, Bianca Redonda and Papa Baya, while Cuarentona and Colorado Ml comprised an intermediate group. These results indicate that variability for salt tolerance exists in this germplasm, and highlights its potential use in breeding programmes. No association was found between the salt tolerance rating and changes in K⁺, Na⁺ and proline accumulation in a salt gradient, thus precluding the use of these parameters as predictors for salt tolerance of these species in in vitro studies.     

Evaluation of salt-tolerant genotypes of durum wheat derived from in vitro and field experiments

The performance of selected salt-tolerant genotypes of durum wheat [Triticum turgidum L. subsp. durum (Desf.) Husn.], derived from field and in vitro assessment methods, was evaluated under greenhouse and field conditions. Eight durum wheat genotypes comprising three salt-tolerant genotypes and one salt-sensitive genotype selected from each of the methods were used. This study was conducted under both saline and non-saline field conditions as well as under greenhouse condition with salinized solution culture at 0 mM (control), 75 and 150 mM NaCl (concentrations) using supplemental Ca²⁺. Days to heading, days to maturity, plant height, number of grains per spike, grain weight per spike, 1000 grain weight, number of spikes per m², grain yield and harvest index were recorded in the field experiments. Plant dry weight, Na⁺, K⁺ and Ca²⁺ accumulated in the hydroponically grown seedlings were measured 20 days after salinity treatments. In spite of the smaller range of genotypes used by the in vitro screening method, tolerant genotypes screened by the in vitro method (ITGs) performed comparably with those of the field-derived tolerant genotypes (FTGs) for grain yield under saline field conditions. Field salinity significantly reduced (P < 0.01) means of all traits averaged on eight tested genotypes. In vitro salt-tolerant genotypes Dipper-6 and Prion-1 produced the highest dry weight and K⁺/Na⁺ ratio under salt stress conditions (150 mM NaCl) in the greenhouse. Although dry matter correlated with the grain yield (R² = 0.37), the regression coefficient was higher for shoot K⁺/Na⁺ ratio (R² = 0.44). Dipper-6 (ITG) and Prion-1 (ITG) genotypes have been ranked superior while Massara-1 (ISG) was inferior for salt tolerance in the regression analysis. However, based on grain yield reduction Ajaia/Hora/Jro/3/Gan (FTG) and PI40100 (ITG) were the most tolerant having 58% and 60% reduction, respectively.     

玉米耐盐基因ZmHKT1的序列变异分析

HKT(High-affinity potassium transporter,高亲和性钾转运蛋白)基因所编码的蛋白能起到维持植物体内的Na+/K+平衡的作用。在作物中HKT蛋白主要通过排出叶片中的Na+,从而提高植物耐盐性。对54份玉米自交系材料的Zm HKT1基因及上游序列进行多态性分析。结果表明,在3 784 bp的序列中,共有194个SNP和32个In Del。大部分变异集中在基因上游序列和内含子区,编码区的变异相对较少,说明编码区在进化和人工选择中承受了更大的选择压力。根据序列变异情况可将54份玉米自交系材料划分为15个单倍型,上游序列主要存在3种单倍型,而基因序列存在9种单倍型。遗传重组和连锁不平衡分析发现,全长序列只有3次重组事件发生,上游序列存在着非常强的连锁不平衡。     

盐胁迫浓度和胁迫时的温度对水稻耐盐性的影响

 在水培条件下，用5种不同浓度的NaCl溶液对20个水稻品种的3~4叶期幼苗进行盐胁迫处理，10 d后考查不同品种单株盐害级别、苗高、茎叶干重和根系Na[sup]+[/sup]/K[sup]+[/sup]含量比等性状。结果表明，0.5%的NaCl胁迫下，品种间的盐害级别和根系Na[sup]+[/sup]/K[sup]+[/sup]含量比差异最大；0.8%的NaCl胁迫下，品种间的相对苗高和相对茎叶干重差异最大。其中韭菜青、农林72、80 85、洞庭晚籼和丁旭稻为强耐盐水稻品种；Pokkali、IR26、小白芒、勐旺谷、明恢63为耐盐或中度耐盐品种。进一步选择8个耐盐性不同的水稻品种，在3种温度条件下进行苗期耐盐性鉴定，发现在相对低温[昼/夜为(23±3)℃/(15±1)℃\]时，水稻生长缓慢，不同品种的盐害级别和根系Na[sup]+[/sup]/K[sup]+[/sup]含量比均较低；在温度较高[昼/夜为(36±1)℃/(26±2)℃\]时，水稻生长加快，不同品种的盐害级别和根系Na[sup]+[/sup]/K[sup]+[/sup]含量比均增高；只有在适温条件[昼/夜为(30±2)℃/(24±2)℃\]下，水稻生长较快，品种间的盐害级别和根系Na[sup]+[/sup]/K[sup]+[/sup]含量比差别最明显。提出在适温条件下，以0.5%或0.8%的NaCl浓度进行水稻品种耐盐性的筛选和耐盐特性的遗传研究比较合适。     

PET fabric supported fixed site carrier membrane for selective metal ion transport

Development of a novel fixed site carrier membrane (FCM), supported by PET fabric for metal ion separation is reported. The membranes were prepared by dipping PET fabric into the methylene chloride solution of Poly(5-vinyl-m-phenylene-m′-phenylene-32-crown-10)_ (P(VCE)), a polymeric metal ion carrier. It was found that the flux of mono-valent metal ion transported across the membrane is significantly differed from each other and the flux decreases in the order Cs⁺>Rb⁺>K⁺>Na⁺>Li⁺ irrespective to the anion except perchlorate anion. It was explained in terms of the stability of the complex, formed by crown ether unit of the P(VCE) and the various metal ions, meanwhile, the lower rate of transport in the presence of perchlorate anion was ascribed to its low hydrophilicity.     

磷对NaCl胁迫下玉米幼苗渗透调节能力的影响

研究不同供磷水平对NaCl胁迫下玉米幼苗有机渗透调节物质和离子含量的影响。结果表明,盐胁迫下低磷处理玉米幼苗叶片中可溶性糖和游离氨基酸增加,根系中显著降低;增加供磷水平,叶片中可溶性糖和游离氨基酸含量下降,根系中含量上升,同时叶片和根系中可溶性蛋白含量增加。磷可降低盐胁迫下玉米幼苗各器官中的Na~+含量,同时增加各器官的K~+、Ca~(2+)和Mg~(2+)含量,降低Na~+/K~+与Na~+/Ca~(2+)比值。磷有助于维持植株的碳氮代谢平衡,促进有机渗透调节物质的运输与分配,改善各器官的离子平衡,增强植株的渗透调节能力,从而缓解盐胁迫带来的伤害。     

外源硅对NaCl胁迫下玉米幼苗渗透调节物质含量影响

采用水培法研究外源硅对盐胁迫下玉米幼苗可溶性糖、游离氨基酸等渗透调节物质含量的影响。结果表明,盐胁迫下适量的增加硅供应能增加叶片和根系中渗透调节物质含量,其中根系中游离氨基酸、可溶性蛋白和可溶性糖含量的增加幅度大于叶片;同时能增加玉米幼苗各器官K+的含量,降低不同部位Na+含量,维持玉米幼苗体内的离子平衡。研究表明,硅参与盐胁迫下渗透调节物质在植物中的运输和分配,适宜浓度的硅能提高玉米耐盐性。     

玉米耐盐基因ZmHKT1;5在烟草中的功能验证

HKT类基因是与植物耐盐性密切相关的一类基因。在作物中HKT蛋白可通过排出Na+来维持植物体内的Na~+/K~+平衡,从而影响植物耐盐性。通过在烟草中过表达玉米ZmHKT1;5基因,验证该基因具有提高植物耐盐性的作用。结果表明,过表达ZmHKT1;5基因的T0代材料即显示出叶片耐盐能力的明显提高;T2代转基因株系种子在含盐培养基上的发芽能力明显强于野生型材料,T2代转基因株系幼苗阶段的耐盐能力也得到了明显的提高。通过比较在盐胁迫后2月龄的转基因材料和野生型材料的生理指标,发现野生型材料中MDA和H_2O_2的含量相较转基因材料发生了更为明显的上升,说明转基因材料中过表达ZmHKT1;5基因有效降低了盐胁迫引起的过氧化物积累。综合转基因验证的结果,证明ZmHKT1;5基因具有提高植物耐盐性的作用。     

Chemical analysis and nutritional assessment of the less known pulses,Vigna aconitifolia (Jacq.) Marechal andVigna vexillata (L.) A. Rich.

The seeds ofVigna aconitifolia andVigna vexillata were analysed for proximate composition, minerals, seed protein fractions, amino acids, fatty acids, and antinutritional factors. Both the pulses were found to be rich sources of proteins and minerals like Ca, Mg, Fe, Zn, and Mn.Vigna aconitifolia seeds exhibited fairly high levels of crude lipid. The most limiting essential amino acids in both the pulses were the sulphur-amino acids, cystine, and methionine. Threonine inV. aconitifolia and phenylalanine and tyrosine inV. vexillata occurred in higher quantities when compared with WHO/FAO requirement pattern. Oleic acid and palmitic acid inV. aconitifolia and linoleic acid and palmitic acid inV. vexillata lipids were found to be the predominant fatty acids. The tannin content of both the pulses was negligible compared with the domesticated legumes. The other antinutritional factors like total free phenols, L-DOPA and haemagglutinating activity were also analysed/assayed.