Colloidal systems in food.
\r\n\t- Traditionally accepted topics related to global health security,
\r\n\t- The impact of human activities and climate change on “planetary health”,
\r\n\t- The impact of global demographic changes and the emergence chronic health conditions as international health security threats.
\r\n\t- A theme dedicated to the COVID-19 Pandemic,
\r\n\t- Novel considerations, including the impact of social media and more recent technological developments on international health security.
\r\n\tThe goal of this book cycle is to provide a comprehensive compendium that will be able to stand on its own as an authoritative source of information on international health security.
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A member of multiple editorial boards and co-author of over 550 publications.",coeditorOneBiosketch:"An Associate Professor of Surgery & Integrative Medicine at Northeast Ohio Medical University and Cardiothoracic Surgeon at the Summa Health Care System. A prolific writer and presenter, with multiple books, hundreds of peer-reviewed articles, and innumerable presentations around the world.",coeditorTwoBiosketch:"A CEO of the INDUSEM Health and Medicine Collaborative, Global Executive Director. of the American College of Academic International Medicine (ACAIM) and head of the World Academic Council of Emergency Medicine.",coeditorThreeBiosketch:"A Director of Research in the Department of Emergency Medicine at Nazareth Hospital in Philadelphia, USA, and co-chief editor of the International Journal of Critical Illness and Injury Science. 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They are multicomponent systems containing different types of ingredients. Therefore, researches about food colloid are important; but first, we must be answered to below questions before starting. (1) How the physical properties of structure, stability, and rheology are influenced by the ingredient composition and formulation conditions? (2) How the interactions between various kinds of dispersed entities (e.g., particles, droplets, and bubbles) and macromolecules (proteins and polysaccharides) influence behavior in bulk fluid phases and at solid and liquid interfaces (air-in-water or oil-in-water)?
For these aims, many types of research are annually carried out for the understanding of how different classes of food ingredients control the physicochemical mechanisms determining overall stability and textural properties. Information on these model systems determined the reliable data about these colloidal systems, but not very reliant, because in the food products numerous components exist which influenced the quality and stability of final products. One of the main objectives of the colloid-based approach is the control of biopolymer interactions with the objective of fabricating well-defined nanoscale structures for controlled destabilization of colloidal systems [1, 2].
It should be noted that based on many papers presented at the 13th European Food Colloids Conference, almost all researches are focusing on four main areas: (i) structure and rheology of protein gels; (ii) properties of adsorbed protein layers; (iii) functionality from protein-polysaccharide interactions; and (iv) oral processing of food colloids. But nowadays, the behavior of dispersed systems within the human digestive systems has emerged as a major topic of research interest. Another outstanding influence on future food colloids research has a strong biomedical emphasis (the topic of controlled release and nutrient delivery) [3]. The development of colloid-based strategies to control delivery of nutrients during digestion in gastrointestinal is very important.
In this chapter, the progress in the field of ingredients, microstructure, and stability of food colloidal systems are discussed. Moreover, the application of new structured functional ingredients for the design of novel colloidal matrix (such as multilayer interfaces, multiple emulsion, gel-like emulsions, and so on) are reviewed. In this further, we will discuss: (i) classification and functions of colloidal systems in food; (ii) types of colloidal systems in food; and (iii) stability of colloidal systems.
A colloid system is a type of mixture in which one part is dispersed constantly throughout another. Colloid systems are usually formed when one part is dispersed through another, but does not combine to form a solution. Therefore, there are many types of colloidal systems that depend on the form of the two parts mixed together.
A colloidal system contains two separate phases: a dispersed phase (or internal phase) and a continuous phase (or dispersion medium). The part which is dispersed is known as the dispersed phase and is suspended in the continuous phase. Colloidal systems in food can be classified into different groups based on the states of matter constituting the two phases. Food colloids are sols, gels, emulsion, and foam. For example, egg white foam is a simple colloid system. Air bubbles (disperse phase) are trapped in the egg white (continuous phase) resulting in a foam. The detailed classification of colloidal systems in food is shown in Table 1 [4].
System | Minor phase | Major phase | Products |
---|---|---|---|
Sol | Solid | Liquid | Raw custard, unset jelly |
Gel | Liquid | Solid | Jelly and jam |
Emulsion | Liquid | Liquid | Mayonnaise, milk |
Solid emulsion | Liquid | Solid | Butter, margarine |
Foam | Gas | Liquid | Whipped cream, whisked egg white |
Solid foam | Gas | Solid | Meringue, bread, cake, ice cream |
Colloidal systems in food.
Food colloids give structure, texture, and mouth-feel to many different food products; for example, jam, ice cream, mayonnaise, etc. Food colloid contains hydrocolloid that provides thickening, gelling, emulsifying, and stabilizing properties in food products [5].
Food hydrocolloids are high molecular weight hydrophilic biopolymers used as functional constituents in the food processing to modify microstructure, texture, flavor, and shelf-life. The term “hydrocolloid” comprises all the numerous polysaccharides that are obtained from plants, seaweeds, and microbial sources, as well as modified biopolymers made by the chemical or enzymatic treatment of starch or cellulose. One of the key functional roles of food hydrocolloids is in the preparation of emulsions and in the control of emulsion shelf-life. Most hydrocolloids can behave as stabilizers (stabilizing additives) of oil-in-water emulsions, but sole a few of them can act like emulsifiers (emulsifying agents). The second functionality needs considerable surface activity at the oil-water interface, and therefore the capability to favor the development and stabilization of fine droplets throughout and next emulsification [6, 7].
The most part of colloids are stable, but the two phases may separate during a period of time due to an increase in temperature or through physical force. Furthermore, they may become unstable after freezing or heating, especially if they contain an emulsion of fat and water. The details of the instability of food colloids are reviewed in further part.
A sol can be defined as a colloidal dispersion in which a solid is the dispersed phase and liquid is the continuous phase. Gravy stirred custard and other thick sauces are some of the examples. While jelly is formed, gelatin is scattered into a liquid and heated to make a sol. As the solution cooks, protein molecules unwind developing a network which traps water and creates a gel.
If corn flour is mixed with water and heated, the starch granules absorb water till they rapture, after that starch granule disperses in the water and the mixture becomes more viscous and makes a gel after cooling. Additional types of gel are formed with pectin and agar. Pectin, a form of carbohydrate found in fruit, is used in the production of jam to help it set. Agar is a polysaccharide extracted from seaweed which is capable of forming gels. If a gel is allowed to stand for a time, it starts to “weep.” This loss of liquid is known as syneresis. The proper ratio of the ingredients is necessary to achieve the desired viscosity of the sols at a certain temperature.
Sols can be transformed into gels as a result of a reduction in temperature. In pectin gels, the pectin molecules are a major phase and the liquid is the scattered phase, whereas in pectin sol, the pectin molecules are a minor phase and the liquid is a major phase. Sols can be made as an initial step in the creation a gel. Jams and jellies produced using pectin are traditional cases that make a sol before the preferred structure.
Numerous natural and processed foods involve either relatively or entirely as emulsions or have been in an emulsified form at some time through their fabrication containing milk, cream, butter, margarine, fruit beverages, soups, batters, mayonnaise, cream liqueurs, sauces, desserts, salad cream, ice cream, and coffee whitener.
Emulsion products exhibit a wide variety of different physicochemical and organoleptic characteristics in appearance, aroma, texture, taste, and shelf-life. The processing of an emulsion-based food product with specific quality features is influenced by the selection of suitable raw materials (e.g., water, oil, emulsifiers, thickeners, minerals, acids, bases, vitamins, flavors, colorants, etc.) and processing situations (e.g., mixing, homogenization, pasteurization, sterilization, etc.).
An emulsion involves two immiscible phases (typically oil or water), with one of the liquids scattered as fine sphere-shaped droplets in the other. A system which contains oil droplets dispersed in an aqueous phase is termed an oil-in-water or O/W emulsion (e.g., mayonnaise, milk, cream, soups, and sauces). A system which involves water droplets scattered in an oil phase is termed as a water-in-oil or W/O emulsion (e.g., margarine, butter, and spreads) [8].
Multiple (or double) emulsions are multipart liquid dispersion systems well-known too as emulsions of emulsions, in which the droplets of one scattered liquid (water-in-oil or oil-in-water) are more scattered in another liquid (water or oil, correspondingly), making W/O/W or O/W/O. The innermost scattered droplets (hereafter called inner droplets or just droplets, while the droplets of the multiple emulsion will be named, for simplicity, the drops) in the multiple emulsion are disconnected from the external liquid phase by a film of another phase.
Although multiple emulsions are an emerging technology, only a few industrial products based on multiple emulsions exist in the marketplace. The main application of multiple emulsions is a protection system for the controlled release of active compounds. In the food industry, W/O/W emulsions are able to increase the solubility of specific active materials, solubilize oil-insoluble ingredients, and serve as protecting liquid reservoirs for molecules sensitive to outside environmental reactivity including oxidation, light, and enzymes, and act as entrapment reservoirs for covering off flavors and odors.
Applications in the cosmetics trade include aqueous preparations that provide a good “feel” and slow release of active materials or flavors, deposition of water-soluble agents onto the skin from wash-off systems. Most applications are related to the pharmaceutical industry, such as enhancing the chemotherapeutic effect of anticancer drugs, drug immobilization, treatment of drug overdoses, and protecting insulin from enzymatic degradation. However, the size of the droplets and the thermodynamic instability is a significant drawback of this technology. It seems that double-emulsion technology can now be applied in various areas, mainly in food, cosmetics, and pharmaceuticals [9, 10].
Emulsions are suggested as carriers of plant antioxidants in food systems that are discussed deeply in Chapter 2. In fact, plant antioxidants due to the natural sources and health-promoting product are very attractive in food science. So, information about the structure of plant antioxidants, degradation of them in food systems, physical and chemical stabilities of these systems are important for study in the future. In Chapter 3, the application of nanoemulsion in food science is discussed. Nanoemulsion is very attracting due to the advances of nanotechnology in the recent years. Nanoemulsion has been applied in functional foods and pharmaceutical industries. Therefore, nanoemulsion production with a novel technique and its stability is very important.
In general, emulsions are thermodynamically unstable and therefore tend to breakdown over time due to various physicochemical mechanisms. Therefore, stabilizers are used in emulsion formulations for improving their long-term stability, such as emulsifiers, texture modifiers, ripening inhibitors, and weighting agents. Emulsifiers (such as small molecule surfactants, phospholipids, proteins, polysaccharides, and other surface-active polymers) are typically amphiphilic molecules that have both hydrophilic and hydrophobic groups on the same molecule.
The most important polysaccharide emulsifiers in food applications are Arabic gum, modified starches, modified cellulose, pectin, and some galactomannans. The role of the emulsifier is to adsorb at the surface of the freshly formed fine droplets and so prevent them from coalescing with the near droplets to form larger droplets again. Mayonnaise is an example of a stable emulsion of oil and vinegar, when egg yolk (lecithin) may be used as an emulsifying agent.
Emulsions are usually formulated by a single type of emulsifier. But, in some cases, the quality and functional properties of emulsions can be improved by using a combination of several emulsifiers rather than the individual alone. Each of them has a unique molecular and physicochemical characteristic that can be applied for modulation the interfacial properties of emulsion droplets.
Novel or improved functional attributes can often be obtained by using emulsifier mixtures rather than single emulsifiers, for example, enhancements in antioxidant activity, flavor encapsulation, nutraceutical delivery, or textural attributes. Due to the increasing demand for clean-label products, utilization of natural emulsifier’s mixtures can be recommended [11]. In addition, the new technique to conjugate proteins with polysaccharide by Maillard reaction arising in the controlled dry heating between the ℇ-amino groups of proteins and the reducing end carbonyl groups of polysaccharides are established. The most remarkable characteristic of the resultant protein-polysaccharide conjugates is the outstanding emulsifying attributes which are preferred in comparison to commercial emulsifiers [12].
Moreover, wet-heating has been adopted to prepare protein-polysaccharide conjugate. Wet-heating mostly shortens the reaction time to only several hours at high temperature and short reaction time limits the Maillard reaction to initial stage to provide better browning control [13, 14].
Foams consist of small bubbles of gas (frequently air) scattered in a liquid, for example, egg white foam. As liquid egg white is whipped, air bubbles are included. The mechanical action leads albumen proteins to unfold and make a network, entrapping the air. If egg white is heated, protein coagulates and moisture is driven off. This creates solid foam, for example, a meringue. Ice cream, bread, and cake are other instances of solid foams.
Recent interest from researches is the application of structural design principles for the fabrication of edible colloids with novel functional properties. This research activity is driven forward by an increasing recognition within the food industry of the value of colloidal systems as delivery vehicles for nutrients and flavor compounds.
In Chapter 5, the role of electrostatic and steric forces in food colloids and their stability are discussed. Based on biopolymer interaction, a combination of protein and polysaccharide functionality for production of the novel biopolymer with enhanced functional properties is deeply studied. Proteins and polysaccharides are two groups of hydrocolloids that are widely used in food formulations simultaneously. These macromolecules are known to play important physicochemical roles, such as thickening, stabilizing, gelling, emulsifying properties, etc., in food products. Interactions between two hydrocolloids play an important role in the structure and stability of processed foods and depend not only on the physicochemical properties of proteins or polysaccharides alone [15–17].
Nowadays, the intelligent manipulation of protein and polysaccharide interactions provides opportunities for the design of new ingredients and interfacial structures with applications in the food and pharmaceutical industries.
So, food scientists can control the microstructure, texture, and shelf-life of edible colloidal systems with attention to theirs. Protein-polysaccharide interactions could play a key role in the nanoscale engineering of novel foods designed to address the widespread health concerns associated with obesity problem and the release of specific nutrients [18].
In addition, nowadays multilayer interfaces are very interested in food industries. Multilayer interfaces in food colloids typically consist of adsorbed layers of proteins and polysaccharides made by the sequential or simultaneous deposition of oppositely charged macromolecules at the surface of emulsion droplets [19].
With the advancement of nanotechnology in different fields such as food industry, some researchers studied various nanoencapsulation techniques for controlled and protection of some bioactive ingredients including pharmaceuticals and food bioactive components with the high bioavailability. Based on the main applied ingredients/equipment for the formulation of encapsulation systems, nanocarriers are classified into five groups: (1) lipid-based nanocarriers (such as nanoemulsions, nanoliposomes, and nanolipid carriers); (2) nanostructured colloids nanocarriers (such as caseins, cyclodextrins, and amylose); (3) nanocarriers produced by special equipment such as electro-spinning/spraying, nanospray dryer, and micro/nanofluidics systems; (4) biopolymers nanoparticles nanocarriers (such as single biopolymer nanoparticles, biopolymer-biopolymer complexation, nanogels of alginates, whey, soy proteins, and chitosan, nanotubes, or nanofibrils); and (5) miscellaneous nanocarriers (such as nanoparticles made from chemical polymers, nanostructured surfactants, inorganic nanoparticles, and nanocrystals). Hence, it is possible to choose appropriate nanodelivery systems based on the solubility and predicted functionality of bioactive components.
In last few years, there has been many published studies on the nanoencapsulation of different food ingredients such as phenolic compounds and antioxidants, natural food colorants, antimicrobial agents and essential oils, minerals, flavors, essential fatty acids and fish oil, and vitamins [20]. Active compounds such as antioxidants and antimicrobials are added into the food formulation for aims of quality loss and microbial safety management. But there are limitations such as pro-oxidation in lipid foods and compliance of regulatory maximum allowable concentration. Therefore, controlled release packaging (CRP) is a novel technology that is applied for the package with release active compounds in a controlled trend to improve safety and quality for food products during storage. Research in controlled release packaging focused on released systems such as active compounds from the package, non-releasing antimicrobials or antioxidants, oxygen absorbers, and free-radical scavengers those grafted on to packaging materials [21].
One developing area in the application of colloidal dispersions is the manufacture of functional foods. Functional foods are becoming progressively favorite among consumers as the result of improved knowledge of functional components and their influence on human wellbeing and biological functions. The customers would like to overcome health problems such as cardiovascular problems and obesity through consuming foods rather than drugs. The plan of functional foods for the delivery of nutraceuticals and micronutrients is a great technological challenge. Colloidal delivery systems are actually found in nature. Casein, for example, is a very illustrative instance of a natural colloidal delivery system for calcium. In milk, calcium is cleverly “engineered” into porous casein colloidal elements of sizes lesser than approximately 500 nm [22]. In Chapter 4, the nanostructured colloids in various areas of food science are discussed.
Nanostructured colloids can be naturally present in food or they can be synthetically manufactured. Some examples of natural nanostructured colloids include casein micelles and β-lactoglobulin in milk, and in the case of synthetically manufactured colloids are metal oxide nanoparticles and clay. Synthetically manufactured nanostructures are added to enhance solubility, improve bioavailability, biologically active compounds protection, increasing shelf-life, color, flavor, and add nutritional value.
The industrial sciences have been of great attention to the development of new bio-based structures with potential in innovative applications. Structures with gel-like behavior are usually used in the cosmetic, pharmaceutical, and food industries for the aim of controlling the physical properties of final products. In the food industry, words like oleogels and organogels have been increasingly used. Oleogels are new emulsion-based structure that can be used to control phase separation and decrease the mobility and migration of the oil phase, providing solid-like properties without using high levels of saturated fatty acids as well as to be a carrier of bioactive compounds. In this area, it can be used as the food grade and bio-based structurants for producing edible oleogels with fat replacement and structure-tailoring functionality [23].
Fruit trees play an important role in human nutrition; and among these highlights, the papaya, this is a crop of tropical climate, widely appreciated for being one of the few fruit that provide continuous production throughout the year after the start of fruiting, to possess fruits with a high nutritional value, and to achieve high yields; generating good income to the families dedicated to their cultivation, due to the high prices that reaches in the market [1].
\nCarica papaya Linn belonging to family Caricaceae is known as papaya in France, United Kingdom, Mexico, Cuba, etc., papita in India, tree melon in Holland, paw paw in Australia and United Kingdom, and mamao in Brazil. The plant is native of tropical America. The properties of papaya fruit and other parts of the plant are also well known in traditional system of medicine. During the last few decades, considerable progress has been achieved regarding the biological activity and medicinal application of papaya and now it is considered as valuable nutraceutical fruit plant [2].
\nThe world production of papaya occurs in more than 60 countries, according to FAO, for the year 2010, a production of 11,568,346 tons of the fruit was registered, with the main producing countries of the highest to lowest volume: India, Brazil, Indonesia, Nigeria, Mexico, Ethiopia, Colombia, Thailand, and Guatemala [3].
\nAs a crop is an important source of employment, has a good yield, earliness to enter into production, and guarantees staggered crops throughout the year. Despite all these advantages, it does not reach the maximum productive potential, this is due in large measure to the problematic that it manifests in terms of the quality of seed [4]. Is consider one of the tropical fruits more appreciated for fresh consumption and for industrialization [5]. In Mexico like in Cuba, Maradol variety is the more cultivated, is a Cuban variety.
\nDifferent varieties of papaya are commercially propagated through seed an easy management and low cost, without taking into account the heterogeneity caused by crossed polinization [5].
\nIn the sowing of papaya, it is best to use freshly harvested seeds, because while increasing the storage time of the seed, the germination rate decreases [6]. The majority part of the papaya sowing is done with stored dry seeds, and in this condition, the seed germination is erratic, asynchronous, slow, and incomplete [7], which diminishes the germination percent [5]. The desiccation produces stress in the papaya seeds when the moisture content lowers to 8.0% [8], being the cause of the seed dormancy or metabolic quiescence [9, 10]. In Taiwan, the papaya industry is limited by seed germination rates [11]. This is attributed to the presence of inhibitors as the phenolic compounds in the sarcotesta and seed coat [12, 13], and in some cases, the seeds lack embryos [6].
\nThe seed is enclosed in a gelatinous sarcotesta (aril or seed coat), which is formed from the outer integument [14]. The sarcotesta can delay germination, and also dormancy is observed in seeds from which the aril has been removed [15].
\nPapaya, like many plants, presents as one of its main problems in its reproduction, the dormancy of the seed which influences the quality of it; because by reaching its maximum point of maturity, it initiates a period of latency produced by internal and external factors. It is normally interrupted when the natural conditions suitable for germination are present or when treatments are used that help to propitiate these ideal conditions and increase the percentages of germination [16], but in vitro conditions favored germination of papaya more than in vivo environment [17].
\nThe seed of papaya is characterized by being bitegumented, since the internal tegument originates the tegumen and the external one to the testa, which is multiplicative up to 60 layers and 3 distinctive strata: endotesta, mesotesta, and exotesta (sarcotesta). This last one of semipermeable consistency, high humidity and concentrate phenolic compounds that, as a whole, induce latency. This causes the inhibition of fluid and gas exchange, delayed dehydration and colonization of pathogenic microorganisms. In [18] are mentioned others researchers who investigated this problem [19, 20].
\nOne of the ways to break the latency of the seeds and make them have a good quality overall, increasing the percentages of germination is using the different methods of scarification.
\nThe methods of scarification include physical, mechanical, and biological treatments such as dry heat, the rupture of the testa, the soaking in water, and chemical solutions that promote the germination of the seeds, where any treatment that destroys or reduces the impermeability is called scarification, so in some cases, it is only enough to destroy a single point of the cover to produce the imbibitions and exchange of gases and thus initiate the germination [21].
\nApparently, latency is a survival mechanism in the presence of certain climatic conditions: very low temperatures, alternations of dry and humid times, and desert climates. The exact causes of the latency phenomenon are unknown, and on the other hand, when the latency is due to testa conditions, the lethargy ends at the moment that it cracks or weakens by mechanical or chemical actions or by effect of the environment [22, 23].
\nDifferent seed treatments to promote germination and to reduce germination time are mentioned in [24], sowing seeds and at warm temperatures, exposing dry seeds to 10°C prior to sowing, drying seeds and soaking seeds in distilled water, potassium nitrate, thiourea, sodium thiosulfate, tannic acid or ferulic acid. The same authors described contradictory results using gibberellins on papaya seed germination. But in your research, they demonstrated that dehydration to 5.3% or 6.9% and 6.8% moisture content, followed by exposure to subzero temperatures and treatment with GA3, were the most favorable combined treatments to enhance papaya seed germination.
\nThe used of smoke water on seed germination and seedling growth of papaya, cultivar Tainung No. 2 consistently and significantly increased the percentage of nitrogen in roots and shoots and significantly increased the percentage of magnesium in shoots. In these experiments, smoke-water showed potent germination promotion at low concentrations and promoted multiple growth attributes such as chlorophyll content and seedling vigor index at all concentrations in papaya seedling production [11].
\nPregerminative treatments are used to break the latency status of the seeds. In [25] are mentioned stratification and scarification. Scarification is any process that breaks, scratches, mechanically alters or softens the covers of the seeds to make them permeable to water and gases.
\nSeed scarification methods have been developed and modified over time to make these more practical and effective. Important methods of seed scarification include heat, freeze–thaw, mechanical, and acid scarification [16].
\nIn the scientific literature, some types of scarification are described, such as mechanical [26, 27, 28], physical [29, 30, 31], chemical [32, 33], and biological. Mechanical, physical, and biological scarification have disadvantages in relation to chemical scarification, because they require more time, are laborious and inadequate to condition large quantities of seed; while chemical scarification still requires more research [18], especially with calcium hydroxide.
\nThere are chemical substances used to scarify seeds, among the most used are the sulfuric acid [34], sodium hydroxide, and hydrochloric acid [7, 18, 20, 31]. The positive effect of the use of NaOH in the benefit of papaya seed is that simulates natural degradation of sarcotesta and improves the conditions of the seed, so it is a viable alternative for use in conditioning seed [18]. Other results have been demonstrated that the combination of NaOH treatment and stratification is an effective practice to break Iris lactea var. chinensis seeds dormancy and improve germination percentage [35].
\nChemistry scarification is considered as one of the most effective scarification methods used for seed scarification. Sulfur acid is the most popular and effective chemical product for acid scarification. The effectiveness of acid scarification depends on concentration of acid duration of scarification and species and cultivars used [16, 36].
\nTraditionally, it has been used to separate the mucilage from the papaya ferment the seeds in water at different time intervals and the sunny one for 2 or 3 h [37].
\nThe objective of the research was to evaluate the scarifying effect of calcium hydroxide on the germination and vigor quality of papaya seeds, Maradol variety.
\nThe experiment was carried out in the Laboratory of Seed Test of the plant of Benefit Manuel Espinosa Ramírez of the business unit of seeds base Granma, belonging to the company producer and marketer of seeds, using seed of papaya ‘Red Maradol’, collected in areas of the Experimental Station Jucaibama of the Agricultural Research Institute Jorge Dimitrov, Bayamo, Granma province.
\nThe sample consisted of 60 randomly selected fruits of hermaphroditic plants in a commercial production lot of approximately 1 ha (2222 plants), showing commercial maturity (two strips), of homogeneous size (±2 kg). The seeds were extracted, and the batch was homogenized; 200 g of fresh dough were deposited with 500 ml plastic flasks representing each experimental unit.
\nThe treatments were composed of the solution of calcium hydroxide (CaOH2) at three doses (60, 80, and 120 g l-1 of water) by dipping the seed for a period of 24 h, the standardized sodium hydroxide (NaOH) at 25% with a 15-min immersion time, for a total of four treatments plus the control and six replicates. The control consisted in fermenting in running water the seed for 24 h.
\nTo eliminate the sarcotesta (aril), the seeds were rubbed between two jute cloths where the time needed to remove all the aril of the material was evaluated, being the optimal time to use of 15 s for each treatment given the amount of material to process. Immediately, they were rinsed three times with running water, spread over a sieve in the shade and at room temperature (28 ± 1°C) for drying for 48 h.
\nThe physical quality of the seed was determined by the effectiveness of the product, physical appearance, and mechanical damage within 3 days of the treatment, compared to the control; the physiological quality was determined by the germination percentage 7 days after sowing (vigor), being valued by the germination rate [38], and 28 days for final germination (as indicated in the germination standard, with the method in sand). The seed was soaked for a term of 24 h and placed in previously disinfested aluminum trays at a temperature of 100°C.
\nThe incubation was carried out in the germinating chamber Paul Polikeit, Model HALLE S. A, with 80% of relative humidity, 40 ± 2°C of temperature and natural light; for the sanitary quality, it was determined by assembling all the treatments with the method between paper (BP), evaluating by observing the evidences of the development of microflora on the seed, during the germination test.
\nTo carry out this test, three repetitions of 25 seeds were used by extraction, it was put to incubated in water the seeds for a time of 24 h, after the time elapsed, each seed was sectioned in longitudinal form leaving the cotyledons visible placed in culture tubes wrapped with aluminum foil adding a solution of 2, 3.5-triphenyl chloride tetrazolium to 1%, and the tubes were placed in an incubator at 35 ± 1°C for a time of 2 h.
\nThe experimental design used was completely randomized with bifactorial arrangement and six replicates. The variables assessed were vigor (vigor, 120, 240, and 360 days of conservation at 4–8°C, in percent); germination (germination, 120, 240, and 360 days of conservation at 4–8°C, in percent); the time needed to eliminate aril (s), mechanical damage, MD (%), and abnormal plants, AP (%), according to ISTA Methodology [38].
\nThe data for each measured variable were statistically processed to check compliance with the normal distribution of the data (Kolmogorov-Smirnov test) and the homogeneity of the variances (Bartlett test). These two premises of the analysis of variance were not met, even after testing several data transformation equations, so we proceeded to the application of nonparametric variance analysis through Kruskal-Wallis, to demonstrate the existence or not of variability between treatments with a probability level of 0.05. The averages (aver.) of the treatments, the standard deviation (sd) of the mean, and the significance are shown. The multiple comparisons between treatments were made through the differences between the averages of the ranges [39].
\nWe also performed analysis of partial correlations between variables with the use of Spearman correlating coefficient, with the aim of determining the existence of linear relationship between selected variables. Those variables that could have a direct or inverse relationship were selected in relation to the vigor and germination in their different times used as the aril, the mechanical damages, and the percentage of abnormal seeds.
\nStatistical processing was carried out with the use of statistical packages MINITAB 13 [40] for the test of homogeneity of variances and Infostat 2017 [41] for the rest of the statistical analyses.
\nSignificant differences were found between the different treatments in the vigor of the seeds (Table 1). The highest level of vigor of the seed was due to the use of calcium hydroxide, more than sodium hydroxide and fermentation. This tendency was maintained during the different storage times of the seeds at a constant temperature. Significant although not shown statistically, there is evidence of a decrease in the seeds vigor for all the evaluated treatments by increasing the conservation time of papaya seeds, which suggests that it is more efficient to apply calcium hydroxide in order to improve the response of papaya seeds with a minimum storage time.
\nVigor (%) | \n|||||
---|---|---|---|---|---|
Compound | \nDose | \n0 days | \n120 days | \n240 days | \n360 days | \n
Aver. ± SD | \nAver. ± SD | \nAver. ± SD | \nAver. ± SD | \n||
Ca(OH)2 | \n60 g l−1 | \n80.7ab ± 2.8 | \n83.7a ± 2.7 | \n77.3b ± 2.3 | \n71.7ab ± 5.7 | \n
Ca(OH)2 | \n80 g l−1 | \n81.7ab ± 1.6 | \n81.2ab ± 1.2 | \n79.3ab ± 2.3 | \n60.0bc ± 6.8 | \n
Ca(OH)2 | \n100 g l−1 | \n89.3a ± 1.2 | \n82.3a ± 2.2 | \n82.3a ± 1.7 | \n75.7a ± 3.6 | \n
NaOH | \n25 g l−1 | \n50.3c ± 4.2 | \n42.7c ± 3.7 | \n26.5c ± 3.4 | \n6.7c ± 0.5 | \n
Fermentation \n | \n75.3bc ± 1.9 | \n75.8bc ± 3.4 | \n73.5bc ± 3.5 | \n71.7ab ± 4.8 | \n
Effect of calcium hydroxide and sodium hydroxide with different doses on the vigor of the seeds in different storage times.
Different letters indicate significant differences to p ≤ 0.05 through the differences between the average of the ranges.
The variability in the response of calcium hydroxide could be a consequence of the fact that some seeds within the same batch have a more persistent dormancy than others and that small and large seeds can be found in the same batch [42].
\nSodium hydroxide reached lower percentages, even less than 60%, which is the minimum value established for Cuba for this crop [43, 44].
\nThe physiological quality of the papaya seed is characterized by a high sensitivity to several factors with respect to germination and vigor, considering that there are integrating elements of great importance at the plantation level [45].
\nFor the germination (Table 2), the application of calcium hydroxide obtained the best results, with percentages over the 80% in comparison with the rest of the treatments. Germination was more affected when the seeds were treated with sodium hydroxide and to the extent that the storage time of the seed was increased. Like the vigor, the germination percentage decreases in the treatments when the conservation time increased.
\nGermination (%) | \n|||||
---|---|---|---|---|---|
Compound | \nDose | \n0 days | \n120 days | \n240 days | \n360 days | \n
Aver. ± SD | \nAver. ± SD | \nAver. ± SD | \nAver. ± SD | \n||
Ca(OH)2 | \n60 g l−1 | \n88.0ab ± 2.18 | \n87.3b ± 1.6 | \n80.2bc ± 1.73 | \n89.3ab ± 6.6 | \n
Ca(OH)2 | \n80 g l−1 | \n89.0ab ± 1.9 | \n89.0ab ± 1.6 | \n81.8ab ± 1.2 | \n70.5bc ± 8.2 | \n
Ca(OH)2 | \n100 g l−1 | \n94.0a ± 2.9 | \n92.3a ± 2.0 | \n85.3a ± 1.8 | \n88.8a ± 4.3 | \n
NaOH | \n25 g l−1 | \n65.3c ± 4.2 | \n56.0c ± 3.2 | \n42.0d ± 3.0 | \n15.5c ± 3.1 | \n
Fermentation \n | \n83.7bc ± 2.3 | \n83.7bc ± 2.7 | \n76.3cd ± 2.9 | \n84.3ab ± 6.08 | \n
Effect of calcium hydroxide and sodium hydroxide with different doses in germination percentage of seeds.
Different letters indicate significant differences to p ≤ 0.05 through the differences between the average of the ranges.
Physiologically, these results could be interpreted as a sequence of events of deterioration that begins with problems of functionality in the seminal membranes, which causes an excessive flow of cellular constituents, evidenced this by the high absorbance values and consequent loss of metabolites, the magnitude of which can restrict the germinative process [46].
\nCold stored papaya seeds maintained significantly higher germination and better seedling vigor than the room stored seeds. With the increase in the duration of storage seed germination decreased after 20 mo. at room temperature, it declined marginally during the same period when kept in cold storage. Irrespective of the storage conditions, seeds kept in sealed polythene bags or plastic bottles had better germination and seedling vigor than those on paper and cloth bags. Shoot length and dry weight decreased significantly with the increase in the duration of storage. Viability of papaya seeds can be maintained considerably at room temperature up to 8 mo. by storing the seed in sealed, preferably airtight, polythene bags or plastic bottles. Cold storage using polythene bags or plastic bottle is recommended [47].
\nThe storage conditions are very important. According to the classification of seed storage behavior, the papaya seed is classified as recalcitrant seed [48], others in intermediate seed [49]. In storage behavior ambient conditions, the papaya seeds survive for a short period of time [50] and are considered intermediate between recalcitrant and orthodox attribute and deteriorate rapidly at higher storage temperatures and relative humidity. Fresh seeds give higher germination rate and seedling vigor that will decline with increasing the storage time [51] and consider that the best conditions for papaya seed storage is when containing 6.0% moisture and stored at 0°C gave higher percentage of germination, lower dormancy, and seed death.
\nIn [18], the treatment that most affected the germination was the application of sodium hydroxide and the higher incidence of microorganisms, with high percentages of plants affected by fungi, which remained even below the germination approved for marketing, which requires, more than 60% [43].
\nIn [52], described some pre-sowing treatments with the finality of increase the germination, like preconditioned papaya seed at 24°C before transfer to 32°C, soaked in KNO3 for 30 min, soaking in gibberellic acid (GA3; 200 ppm) for 24 h resulted in highest germination percentage in soil compared to α-naphthalene acetic acid (NAA) and KNO3 treatment, and using a protocol of sterilization and germination of papaya seeds in response to light emitting diodes and got 100% of sterilization and 100% of germination.
\nThe efficiency of the different scarification methods has been demonstrated in several investigations to favor the germinative process. In [53] were evaluated several methods of scarification of pacain seeds (Chamaedorea sp.), where the germination results indicated that the highest percentage was 77.8% and that corresponded to cold water treatment for 15 days (physical scarification), followed by treatment with mechanical scarification by hammer blow with 77.17% compared to the absolute control that obtained a 5.67% of germination.
\nWith chemical scarification methods also obtained positive results [54], and verified germination percentages of 97 and 94%, respectively, in seeds of Centrosema macrocarpum for 10 min immersion.
\nThe treatments used release the aril at different time intervals and there are differences between them in relation to the mechanical damage and the quantity of abnormal seeds of the Maradol variety (Table 3). The best results were obtained when calcium hydroxide was applied at the 100 l−1 dose and with sodium hydroxide. Those compounds only needed an average time of 12 s to achieve an efficiency in the detachment of the aril, while with the application of the control dose, the worst results were obtained, with an average of 38 s that in some cases reached reaching more than 420 s to achieve the release of the aril, so that with this result, it is inferred that to work 25 kg of wet seeds, it would take approximately 38 min if treated with sodium hydroxide or calcium hydroxide in 100 l dose and approximately 106 min for the case fermented only with water.
\nCompound | \nDose | \nAril (s) | \nMD (%) | \nAP (%) | \n
---|---|---|---|---|
Aver. ± SD | \nAver. ± SD | \nAver. ± SD | \n||
Ca(OH)2 | \n60 g l−1 | \n21.2cd ± 1.3 | \n1.0a ± 0.9 | \n1.5ab ± 1.5 | \n
Ca(OH)2 | \n80 g l−1 | \n17.7cd ± 2.5 | \n1.0a ± 0.9 | \n0.7a ± 1.0 | \n
Ca(OH)2 | \n100 g l−1 | \n13.3ab ± 1.4 | \n0.5a ± 0.6 | \n0.4a ± 0.7 | \n
NaOH | \n25 g l−1 | \n12.0a ± 0.9 | \n0.5a ± 0.2 | \n3.3b ± 1.0 | \n
Fermentation \n | \n37.7d ± 6.3 | \n9.2b ± 1.2 | \n0.8a ± 0.9 | \n
Effect of calcium hydroxide and sodium hydroxide with different doses in the time (s) of detachment of the seed’s aril.
Different letters indicate significant differences to p ≤ 0.05 through the differences between the average of the ranges.
When carrying out an essay [55], with different methods to improve the germination of four forage shrubs legumes [Tagasaste (Cytisus proliferus) and three species of Teline (Genista)], obtained the most encouraging results for the elimination of the aril in the treatment with concentrated sulfuric acid for 30 min, demonstrating the effectiveness of chemical compounds in the scarification of seeds.
\nRefs. [7, 20] observed a negative effect on the papaya germination due to the presence in the aril or sarcotesta of inhibitory substances. It is also said that the marked decrease in germination, in the presence of sarcotesta is due to the low oxygenation of the seeds, which is why it is recommended to remove it. Likewise, in a study carried out in Honduras with seeds of the papaya, Maradol variety, higher percentages of germination were obtained with freshly harvested and oared seeds (83%), while fresh seeds with burned arils showed a lower percentage of germination, with 75% [56].
\nSimilar results presented by [18, 20] which treated seeds with very corrosive products not only the germination is affected, but that several plants emerged with problems fundamentally in the radicle and hypocotyl, assuming the influence of other factors such as fluctuation in the moisture of the seed in the time of conservation.
\nThe variables that were selected for the multiple correlation analyses (Table 4) showed that the release of aril seed correlated significantly with germination and vigor 360 days, positively and in a mean size. No correlations were found between this variable and the rest of the values of germination and vigor evaluated. Candiani et al. [57] concluded that the germination of Michelia champaca L. seeds is hindered probably due to the presence of inhibiting substances in the aril, is considered as endogenous causes of seed dormancy which include factors such as phytohormones, or by interference with water uptake [58]. Abscisic acid (ABA) is the key inhibitor of germination in Taxus yunnanensis seeds during wet sand storage [59].
\nCorrelations analysis | \n|||
---|---|---|---|
\n | Aril | \nMD | \nAdnor | \n
Ger 0 | \n0.13 ns | \n0.07 ns | \n−0.74* | \n
Ger120 | \n0.24 ns | \n0.15 ns | \n−0.71* | \n
Ger240 | \n0.25 ns | \n0.18 ns | \n−0.72* | \n
Ger360 | \n0.46* | \n0.34 ns | \n−0.70* | \n
Vigor | \n0.13 ns | \n0.08 ns | \n−0.73* | \n
Vigor120 | \n0.28 ns | \n0.19 ns | \n−0.70* | \n
Vigor240 | \n0.30 ns | \n0.22 ns | \n−0.69* | \n
Vigor360 | \n0.46* | \n0.34 ns | \n−0.70* | \n
Spearman correlations coefficient between variables.
Indicate significant differences to p ≤ 0.005.
Ger is germination, MD is mechanical damage, and adnor is adnormal plants.
The mechanical damage caused to the seeds during this process in this experiment did not correlate with the different levels of germination and vigor that were studied; however, the percentages of abnormal plants in the different time intervals evaluated showed a significant correlation, with medium to high values, but inverse and indicates that as the percentage of abnormal plants increases in a seed lot, the number of seeds germinates decreases and the vigor. Germination vigor is driven by the ability of the plant embryo, embedded within the seed, to resume its metabolic activity in a coordinated and sequential manner.
\nWas analyzed the vigor tests on lettuce (Lactuca sativa L.) seeds and their correlation with emergence [60], demonstrated that saturated salt accelerated aging and digital image analysis were the best laboratory tests for lettuce seed vigor evaluation, especially for seed lots to be used for plug seedling production. In some case, the use of seed vigor tests is used to predict field emergence in plants, like lucerne (Medicago sativa) [61].
\nIn studies [62] about studied the correlations of seed germination percent of two sweet corn hybrids (Zea mays L.) with field emergence and some measured traits related to yield, the results of correlation analysis indicated that there was a high positive correlation between seed germination ability and vigor with seedling field emergence and most of the measured traits, as the percent of the radicle emergence.
\nThe germination vigor depends on multiple biochemical and molecular variables. Their characterization is expected to deliver new markers of seed quality that can be used in breeding programs and/or in biotechnological approaches to improve crop yields [63].
\nCalcium hydroxide has great potential to be used as biocide in agriculture, because it has the advantage of not being phytotoxic, is economic and easy to use and is harmless to the environment and to humans. Decreased Ca levels in the nutrient medium reduced soybean leaf dry matter during seed fill, seed production, seed Ca concentration, and seed germination and increased the incidence of seedling disorders such as watery hypocotyl and epicotyl necrosis [64].
\nIn [65] performed standard germination tests, germination and growth rate, accelerated aging, electrical conductivity, respiration rate and ATP content, to evaluate the vigor of the seeds of Bromus biebersteinii Roem & Schult and determined that the correlations with the behavior in the field of that forage were accelerated aging, respiration rate, and ATP content. The cited authors conclude that a vigor test alone is not adequate to measure the quality of seed lots. A combination of tests which measure both physiological and biochemical aspects should be used.
\nSome researches [66] showed that seed size is an important factor for germination and seedling vigor, establishing that larger seeds produce more vigorous seedlings but with slower emergence. So it can be argued that by using the largest scarified seeds that we have, we can help considerably to decrease the percent of abnormal plants and at the same time increase the vigor and the percentage of germination, variables evaluated in the present investigation.
\nTo correlate characters related to seed germination, is important to investigate the effects of environmental factors prevailing during seed maturation under controlled conditions to understand exact reasons for unusual seed dormancy and germination requirements, for example, the germination of Citrullus colocynthis (Cucurbitaceae) is very sensitive to light and incubation temperature as well as to the environmental conditions associated with the time of seed maturation [67] and seed dormancy is a temporary failure of a viable seed to complete germination under normally favorable physical environmental conditions [33].
\nSeed germination tests assess the ability of the seed to produce a healthy plant when placed under favorable environmental conditions. Germination tests are conducted for a prescribed time period under laboratory conditions that assure optimum moisture, temperature, and light. Unfortunately, these conditions are seldom encountered in the field, and field emergence may be overestimated by standard germination tests. Seed lots that have low germination also are less vigorous due to seed deterioration. As seeds deteriorate, loss of vigor precedes loss of viability, so seeds with low germination usually will be less vigorous. Hence, in seed lots with poor germination, those seeds that do germinate often produce weaker seedlings with reduced yield potential. However, some species (such as many native grasses) have inherently low germination potential and cannot be assumed to have poor vigor due to low germination. Seed vigor usually cannot be assessed by the consumer. Germination is a good indicator of seed vigor [63, 68].
\nUsing efficient methods to scarify papaya seeds can increase the germination percentages of seeds, only if there is good control of environmental factors, because papaya seed germination is affected by light, temperature, oxygen, pH, and the moisture of the substrate [7].
\nIt is necessary to conduct researches about the biometric and morphological characteristics of fruit and seeds, aiming at the maximum germination capacity and seed vigor [69], because biometric studies of seeds and their phenotypical correlations allow the quantitative evaluation of a character’s relevance in relation to another [70]. It continues to investigate the correlations between the different indicators that can characterize the quality of the seeds. In adaptive correlations between seed size and germination time [71], present a model for the coevolution of seed size and germination time within a season when both affect the ability of the seedlings to compete for space and show that even in the absence of a morphological or physiological constraint between the two traits, a correlation between seed size and germination time is nevertheless likely to evolve.
\nSeed germination is a complex process and we need to understand the underlying molecular, hormonal, and mechanical aspects [72]. The environment during seed production has major impacts on the behavior of progeny seeds [73]. For that reason, the seed biology is considered the principal research topic for food security take into consideration the climate change [72].
\nNowadays, there are advances in the propagation of papaya by biotechnological methods. Efficient micropropagation of papaya has become crucial for the multiplication of specific sex types of papaya and in the application of genetic transformation technologies. Significant progress has been achieved using organogenesis and somatic embryogenesis as the shoot tip, axillary bud and single node culture, organogenesis, anther and ovule culture, and regeneration from protoplasts, callus induction and somatic embryogenesis and the mass propagation by ex vitro rooting and acclimatation [74].
\nIn natural conditions, the germination of papaya seeds has difficult by the presence of aril (sarcotesta) that become in a physical barrier which limits the diffusion of water and gases into the seeds and by the effect of phytohormones which preventing germination of seeds, causing dormancy, limiting the development of the embryos and causes a low and variable germination affecting the final percentage. This problem can be solved with the scarification of papaya seed. The results of this research showed that the dormancy by the presence of aril is produced in the papaya seed can be broken with the use of NaOH; but higher results were achieved with the use of calcium hydroxide, Ca(OH)2. The results suggested that chemical scarification with calcium hydroxide can improve germination percentage and vigor of the papaya seeds, take into account that the seed is considered the major essential input in crop production. If the seed quality parameters (vigor, germination rate) decrease, the yields are affected. The scarification of papaya seeds with the use of calcium hydroxide for its proven effect in this research on the benefit of papaya seeds and easy acquisition and will reduce the costs of seed. But a good germination of the papaya seedlings depends on the many environmental factors and excellent agronomy practice. Finally, the effectiveness of scarification methods could change among cultivars within the papaya specie.
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