Antioxidant and radioprotective properties of commercially grown litchi (Litchi chinensis) from India

In an earlier study, radiation treatment (0.5 kGy) and low temperature (4 °C) storage were found to extend the shelf life of litchi fruit var. ‘Shahi’ and ‘China’ from India up to 28 days, while maintaining its physical, biochemical and organoleptic properties. The current study was aimed to elucidate the antioxidant and radioprotective properties of the litchi fruit, and the effect of radiation processing on these properties. The litchi fruit was found to be significantly rich in antioxidant and radioprotective properties, and the antioxidant parameters were found to be well correlated with the phenolic and flavonoid contents. A significant protection to pBR322 plasmid DNA and Escherichia coli cells from radiation induced damage was observed in the presence of litchi juice. Plasmid DNA was well protected even at the dose of 5 kGy, whereas, bacterial cells could be protected up to 0.5 kGy. Neither of these properties was found to be sensitive to radiation processing or low temperature storage.     

Functionality of phosphorylated vicilin exposed to chemical and physical agents

The effects of phosphorylation with sodium trimetaphosphate (STMP) on functional and physicochemical properties of pea vicilin, as probed by high hydrostatic pressure and chemical denaturation were evaluated. The isoelectric point of unmodified and phosphorylated vicilin decreased in the presence of 0.5 M NaCl, resulting in a decrease of the solubility at pH 1.0. The gelation capacity of unmodified vicilin in the presence of NaCl decreased approximately 80% when compared with unmodified vicilin without NaCl. Increasing pressure from 0.1 MPa (atmospheric pressure) to 240 MPa significantly decreased the solubility of vicilin phosphorylated with 4% STMP at pH 1.0 and 4.0 by about 30%. Pressure had no effect on solubility of native vicilin. Pressure treatment at 240 MPa improved the gelation capacity of vicilin phosphorylated with 1% STMP. Glycerol decreased the gelation capacity of vicilin and its solubility in the acidic pH range.     

Physical and mechanical properties of peanut protein films

The properties of peanut protein films were modified using physical and chemical treatments, and their effects on color, mechanical strength, water solubility and barrier to water vapor and oxygen of the films were investigated. Physical treatments consisted of heat denaturation of film-forming solution for 30 min at 60°C, 70°C, 80°C and 90°C, ultraviolet irradiation of films for up to 24 h, and three ultrasound processes of film-forming solution. Chemical treatments consisted of addition of aldehydes and anhydrides. Heat curing at 70°C, ultraviolet irradiation for 24 h, ultrasound for 10 min in a water-bath, and formaldehyde and glutaraldehyde addition caused a significant increase in the tensile strength of the films. The water vapor permeability (WVP) and oxygen permeability (OP) of the films decreased after heat denaturation and aldehyde treatment. OP also decreased with UV treatment. Heat curing was the most effective treatment, making the films stronger, more resistant to water and less permeable to water vapor and oxygen.     

Factors affecting instant properties of powdered cocoa beverages

The increasing number of small and medium scale manufacturers of powder cocoa beverages (PCBs) in Nigeria requires relevant technical data useful in designing new and value added products from cocoa powder. This paper reports a preliminary study carried out to determine how some physical and chemical factors relate to the instant properties of some commercial samples of PCBs produced in Nigeria. The levels of chemical component like moisture, fat and sugar in the PCBs varied from 0.8% to 3.6%, 2.0% to 10.4% and 52.4% to 90.5%, respectively. Physical properties like the bulk density, angle of repose, average particle size and uniformity index of the products varied from 0.49 to 0.81 g/cm³, 25.0° to 37.7°, 0.031 to 0.796 mm and 6.25 to 7.44, respectively. Instant properties such as wettability, dispersibility and solubility ranged from 10.7 to 21.7 s, 50.0% to 94.5% and 44.2% to 76.6%, respectively. These properties differed significantly (p < 0.05) among the 10 commercial samples of PCB studied. Sugar (sucrose) content of products had the most significant (p < 0.05) main effect on their instant properties followed by the fat content. Wetting time showed a significant (p < 0.05) negative linear correlation with sugar content. Agglomeration increased the average particle size, which correlated negatively with uniformity index. The instant properties of fine PCBs (average particle size <0.294 mm) were more predictable than the agglomerated samples.     

The water vapour permeability,mechanical properties and solubility of fish gelatin–chitosan films modified with transglutaminase or 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) and plasticized with glycerol

The effect of glycerol on the mechanical and water barrier properties, as well as on the water solubility, of fish gelatin–chitosan films (4:1, w/w) cross-linked with TGase or EDC was determined. The addition of glycerol in concentrations up to 30% (of the substrate mass) to the fish gelatin–chitosan films modified with TGase or EDC did not change their solubility in buffers of pH 3 and 6 at 25 °C or during heating at 100 °C for 60 min. The chemical and enzymatic cross-linking of the components did not increase the water barrier properties of the films. WVP of the films modified with EDC and TGase was not affected by glycerol at concentrations up to 25% of the substrate mass. Tensile strength of the films decreased after modification of the components with TGase or EDC, respectively, by about 25% and 40%. The elongations of the enzymatically modified films containing 20% of glycerol and of chemically modified films containing 15% of glycerol were, respectively, about 8 and 13 times higher than those of unplasticized films; however, the tensile strengths of plasticized films were, respectively, 2.5 and 5 times lower.     

Empirical,thermodynamic and quantum-chemical investigations of inclusion complexation between flavanones and (2-hydroxypropyl)-cyclodextrins

The inclusion complexation of (2-hydroxypropyl)-cyclodextrins with flavanones was investigated by phase solubility measurements, as well as thermodynamic and quantum chemical methods. Inclusion complexes were formed between (2-hydroxypropyl)-α-cyclodextrin (HP-α-CD), (2-hydroxypropyl)-β-cyclodextrin (HP-β-CD), (2-hydroxypropyl)-γ-cyclodextrin (HP-γ-CD) and β-cyclodextrin (β-CD) and four flavanones (naringenin, naringin, hesperetin and dihydromyricetin) in aqueous solutions and their phase solubility was determined. For all the flavanones, the stability constants of their complexes formed with different CDs followed the rank order: HP-β-CD (MW 1540) > HP-β-CD (MW 1460) > HP-β-CD (MW 1380) > β-CD > HP-γ-CD > HP-α-CD. Experimental results and quantum chemical calculations showed that the ability of flavanones to form inclusion complex with (2-hydroxypropyl)-cyclodextrins was determined by both the steric effect and hydrophobicity of the flavanones. For flavanones that have similar molecular volumes, the hydrophobicity of the molecule was the main determining factor of its ability to form inclusion complexes with HP-β-CD, and the hydrophobicity parameter Log P is highly correlated with the stability constant of the complexes. Results of thermodynamic study demonstrated that hydrophobic interaction is the main driving force for the formation process of the flavanone–CD inclusion complexes. Quantum chemical analysis of the most active hydroxyl groups and HOMO (the highest occupied molecular orbital) showed that the B ring of the flavanones was most likely involved in hydrogen bonding with the side groups in the cavity of the CDs, through which the inclusion complex was stabilised.     

Chemical analysis and preliminary toxicological evaluation of Garcinia mangostana seeds and seed oil

The chemical analysis and preliminary toxicological evaluation of Garcinia mangostana seeds and seed oil have been investigated in order to determine the possibility of using them for human and/or animal consumption. Proximate analysis showed that the seeds had high amount of carbohydrate and were rich in oil (21.68 ± 6.18%) but have a low protein content. The physical properties of the oil extracts showed the state to be liquid at room temperature (25 ± 1 °C) and the colour of the oil golden-orange. The specific gravity of the oil was 0.98 ± 0.01. Among the chemical properties of the oil extracts, acid value, saponification number, iodine value, percent free fatty acid and peroxide value compared well with those of conventional edible oils. The seed flour was found to be a good source of minerals. It contained considerable amounts of potassium (7071 mg/kg), magnesium (865 mg/kg) and calcium (454 mg/kg). Fatty acid composition of the seed oil indicated that the oil contained one essential fatty acids small proportions: linoleic acid (1.30%). The most prevalent fatty acids were palmitic acid (49.5%) and oleic acid (34.0%). Weanling albino rats appeared to suffer no toxicological effects when fed with G. mangostana seed oil in their diet for 8 weeks. Weekly monitoring of the rats showed good physical appearance and steady weight increase. Histological examination of sections of the heart, liver, kidney, spleen and lung revealed that the kidney of some of the rats had some degrees of pathology which included diffuse glomerular and tubular degeneration. No lesion was found in the heart and liver of the rats. The seed oil could be useful as an edible oil and for industrial applications.     

Effect of gamma radiation and cold storage on chemical and organoleptic properties and microbiological status of liquid egg white and yolk

This investigation aims to establish a feasible radiation dose for treating liquid egg white (LEW) and yolk (LEY) at room temperature to improve their microbial safety. Samples of LEW and LEY were subjected to gamma irradiation doses of 0,1,2,3 and 4 kGy at room temperature followed by storage at 4 ± 1 °C. Then the effects of irradiation and cold storage on proximate composition, pH, soluble protein and free sulfhydryl content (SH) were determined for LEW and LEY in addition to the contents of total carotenoids in LEY. Moreover, free fatty acids (FFA) and peroxide value (PV) were determined for lipids of LEY.The microbial safety of LEW and LEY was established during storage throughout the enumeration of the total plate count, enterobacteriaceae, Staphylococcus aureus as well as the detection of Salmonella. The effects of irradiation at 3 kGy dose, which was enough for improving the microbial safety of samples, on amino acid composition of LEW and LEY and fatty acid profiles of LEY lipids were studied. In addition, sensory evaluation was carried out for liquid and scrambled egg white and egg yolk samples. The results showed that gamma irradiation and refrigerated storage had no significant effects on proximate composition and pH of liquid egg samples, while significantly decreased the contents of total carotenoids in LEY samples. Furthermore, gamma irradiation had no significant effects on protein solubility and the contents of free SH in LEW, while induced significant slight decreases in protein solubility and the contents of free SH in LEY. Cold storage, however, showed no significant effects on protein solubility and free SH in all liquid egg samples. FFA contents and PV of LEY lipids significantly increased post irradiation treatments and during storage, but the observed values were relatively low and acceptable. In addition, gamma irradiation at 3 kGy dose had no significant effects neither on the amino acid composition of LEW and LEY nor on fatty acid profiles of LEY lipids. The sensory preference did not alter neither for the liquid egg samples nor for scrambled egg samples that prepared from irradiated liquid egg products. Finally, gamma irradiation at 3 kGy dose appeared to be the optimum for treating LEW and LEY at room temperature followed by cold storage at 4 ± 1 °C.     

Effect of storage time and temperature of milk protein concentrate (MPC85) on the renneting properties of skim milk fortified with MPC85

This study investigated the effect of storage temperature (20–50 °C) and time (0–60 days) on the renneting properties of milk protein concentrate with 85% protein (MPC85). Reconstituted skim milk was fortified with the MPC85 (2.5% w/w) and the renneting properties of the skim milk/MPC85 systems were investigated using rheology. It was found that the final complex modulus (final G∗) and the yield stress of the rennet-induced skim milk/MPC85 gels decreased exponentially with storage time of the MPC85 for storage temperatures greater than 20 °C, with a greater effect at the higher storage temperatures. Changes in the solubility of MPC85 with storage time were correlated with the rheological properties. The primary phase of renneting (cleavage of κ-casein) was not affected by the storage of the MPC85; hence the effect was related to the secondary stage of renneting (aggregation/coagulation of rennet-treated casein micelles). Using a temperature–time superposition method, a master curve was formed from the final G∗, yield stress and solubility results. This suggested that the same physical processes affected the solubility and rennet gelation properties of the milks. It is proposed that the MPC85 protein in rennet-treated skim milk/MPC85 solutions may transform from an interacting material, when solubility is high, to an inert or weakly interacting material, when solubility is low, and that this results in the reduced final G∗ and yield stress of the rennet gels when MPC85 is stored at elevated temperatures for long periods.     

Surface tension and refractive index of guar and tragacanth gums aqueous dispersions at different polymer concentrations,polymer ratios and temperatures

Surface tension and refractive index of guar gum (GG) (up to 0.97% w/w) and tragacanth gum (TG) (up to 1.41% w/w) aqueous dispersions were determined for systems with only one and both gums (mixtures) (up to 1.25% w/w). Experimental determinations were carried out from 5 to 30 °C for surface tension and from 10 °C to 40 °C for refractive index. For mixtures, the effect of polymer ratio (1:3, 1:1 and 3:1) was analyzed at different total gum concentration and temperature. GG showed lower surface activity and refractive index than TG at the same experimental condition. Models for the estimation of surface tension and refractive index as function of gum concentration and temperature were proposed and satisfactorily tested. Both physical and optical properties, in the tested gum concentration range, allowed the determination of different behaviour for both gums because of a critical aggregation concentration for TG dispersions while this phenomenon was not observed for GG.     

Influence of salt and pH on the solubility and structural characteristics of transglutaminase-treated wheat gluten hydrolysate

Hydrolyzed wheat gluten (GH, 77–85% protein) was prepared by limited chymotrypsin digestion at 37 °C for 4 h (degree of hydrolysis = 6.4%) and 15 h (degree of hydrolysis = 10.3%). Microbial transglutaminase (MTGase) treatment (55 °C for 1 h, or 5 °C for 18 h) effect on the solubility and structural characteristics of GH was examined under selected food processing conditions (pH 4.0–7.0, 0–0.6 M NaCl). The MTGase treatment increased solubility of GH by 3–29-fold (P < 0.05) within pH 4.0–7.0. Addition of 0.6 M NaCl or changing the conditions of MTGase incubation did not significantly alter solubility characteristics of GH. The MTGase treatment decreased surface hydrophobicity, and increased carboxyl groups in GH, suggesting cross-linking and deamidation. Fluorescence and UV spectra attributed the improved GH solubility to MTGase-induced polar environment, and partial masking of some nonpolar aromatic amino acids possibly due to high-molecular-weight polypeptides formed.     

Effects of plasticizers, pH and heating of film-forming solution on the properties of pea protein isolate films

Effects of glycerol (3-7% w/w) and sorbitol (4-8% w/w) concentration, pH (7.0, 9.0, 11.0) and heating (90 °C, 20 min) of film-forming solution (FFS) on the water vapor permeability (WVP), moisture content (MC), solubility, light transmission and transparency of pea protein isolate (PPI) films were investigated. Films plasticized with sorbitol exhibited significantly lower WVP, lower MC and higher solubility, in comparison with glycerol-plasticized films. Increasing glycerol content of the films led to increases in WVP and MC but did not affect film solubility. In contrast, increase in sorbitol content had no effect on permeability and MC but resulted in increased film solubility. Moisture sorption isotherms of PPI films suggested that the difference in WVP observed among films plasticized with glycerol and sorbitol might be due to the different hygroscopicity of these plasticizers. The pH of FFS did not have a significant effect on WVP and MC. Solubility of PPI films formed from non-heated FFS was not affected by pH, whereas solubility of films formed from heat-treated FFS generally increased when pH was increased from 7.0 to 11.0. Heating of FFS resulted in improved film transparency. All tested films were characterized by excellent ability to absorb UV radiation. Microstructural observation by scanning electron microscopy did not show differences between sorbitol- and glycerol-plasticized films.     

Changes in the physical properties,solubility, and heat stability of milk protein concentrates prepared from partially acidified milk

A limiting factor in using milk protein concentrates (MPC) as a high-quality protein source for different food applications is their poor reconstitutability. Solubilization of colloidal calcium phosphate (CCP) from casein micelles during membrane filtration (e.g., through acidification) may affect the structural organization of these protein particles and consequently the rehydration and functional properties of the resulting MPC powder. The main objective of this study was to investigate the effects of acidification of milk by glucono-δ-lactone (GDL) before ultrafiltration (UF) on the composition, physical properties, solubility, and thermal stability (after reconstitution) of MPC powders. The MPC samples were manufactured in duplicate, either by UF (65% protein, MPC65) or by UF followed by diafiltration (80% protein, MPC80), using pasteurized skim milk, at either the native milk pH (~pH 6.6) or at pH 6.0 after addition of GDL, followed by spray drying. Samples of different treatments were reconstituted at 5% (wt/wt) protein to compare their solubility and thermal stability. Powders were tested in duplicate for basic composition, calcium content, reconstitutability, particle size, particle density, and microstructure. Acidification of milk did not have any significant effect on the proximate composition, particle size, particle density, or surface morphology of the MPC powders; however, the total calcium content of MPC80 decreased significantly with acidification (from 1.84 ± 0.03 to 1.59 ± 0.03 g/100 g of powder). Calcium-depleted MPC80 powders were also more soluble than the control powders. Diafiltered dispersions were significantly less heat stable (at 120°C) than UF samples when dissolved at 5% solids. The present work contributes to a better understanding of the differences in MPC commonly observed during processing.     

Physical and chemical properties of ultrasonically, spray-dried green banana (Musa cavendish) starch

Ultrasonic wave propagation and the spray dryer technique were applied to study their effect on the physical and chemical properties of green banana starch. The results showed high resistant starch content, which was reduced by ultrasound treatment and also by spray drying. Both techniques increased the solubility, swelling power and water absorption capacity. The gels exhibited non-Newtonian shear-thinning behavior, since flow behavior index was less than one (n < 1). Ultrasound wave propagation reduced yield stress and consistency coefficient in starch gels. Under oscillatory shear, all gels exhibited solid-like viscoelastic behavior, storage modulus was higher than loss modulus to entire frequency range (G′ > G′′), which was confirmed by the Cox-Merz experiment that showed that the complex dynamic viscosity was greater than the apparent viscosity in all samples. The gelatinization temperature was mainly influenced by drying technique and ultrasound treatment reduced the amount of energy required to gelatinize the starch.     

Reducing cracking and breakage of soybean grains under combined near-infrared radiation and fluidized-bed drying

Near-infrared radiation was combined with fluidized-bed drying to reduce the cracking and breakage of soybean grains because of its advantages such as a decrease in the moisture gradient and the stresses development within the grain kernel, leading to high quality of product. Physical qualities, i.e. cracking, breakage and colour, were investigated together with microstructure of soybean grains. Protein solubility and urease activity were also determined. The parameters studied were near-infrared radiation powers of 4, 6 and 8 kW, air velocity of 4.5 m/s, air temperature of 40 °C and grain bed depth of 6 cm. Results showed that the drying rate was increased with the increase of near-infrared radiation power. The moisture content was reduced from an initial moisture content of 20% d.b. to the final moisture contents of 13.5% d.b. (4 kW), 12.8% d.b. (6 kW) and 12.5% d.b. (8 kW). The cracking and breakage of soybean grains occurred negligible, which was lower than 4.4% and 5.3% for cracking and breakage, respectively. At near-infrared radiation powers of 4 and 6 kW was accepted for both soybean grains trade and animal feed industries in Thailand. The total colour difference changes (ΔE) were varied in the range of to 2.9-4.2. The protein solubility of final product was in an acceptable range (77-82%) for feed meal. The reduction of residual urease was varied in the range of to 39-70%. This study has demonstrated that the soybean grains under combined near-infrared radiation and fluidized-bed drying was properly treated.     

Chemical composition and functional properties of raw and roasted Nigerian benniseed (Sesamum indicum) and bambara groundnut (Vigna subterranean)

Benniseed and bambara groundnut seeds were roasted at 80 and 120 °C for 10–60 min. For both flours, the effects of roasting temperature and time on selected functional properties and chemical composition were determined, as were the effects of pH on the emulsification capacity and nitrogen solubility. The chemical constituents of the raw flours were present at higher concentrations than those of the roasted flours except for fat and ash. Protein concentrates of both flours contained 80.5–81.5% crude protein as the major constituent. Nitrogen solubility was lowest at pH 4.0 for raw and roasted benniseed flour and pH 5.0 for raw and roasted bambara groundnut flour. Roasting generally lowered the nitrogen solubility and increased the water and oil absorption capacities while decreasing the foaming capacity and emulsification capacity of both flours.     

High pressure induced changes on sarcoplasmic protein fraction and quality indicators

The combined effect of pressure and mild temperature treatments on bovine sarcoplasmic proteins and quality parameters was assessed. M. longissimus dorsi samples were pressurised in a range of 200–600 MPa and 10–30 °C. High Pressure Processing (HPP) induced a reduction of protein solubility (p < 0.001) compared to non-treated controls (NT), more pronounced above 200 MPa. HPP at pressures higher than 200 MPa induced a strong modification (p < 0.001) of meat colour and a reduction of water holding capacity (WHC). SDS–PAGE analysis demonstrated that HPP significantly modified the composition of the sarcoplasmic protein fraction. The pressurisation temperature mainly affected protein solubility and colour; a smaller effect was observed on protein profiles. Significant correlations (p < 0.001) between sarcoplasmic protein solubility and both expressible moisture (r = −0.78) and colour parameters (r = −0.81 to −0.91) suggest that pressure induced denaturation of sarcoplasmic proteins could influence to some extent WHC and colour modifications of beef. Changes in protein band intensities were also significantly correlated with protein solubility, meat lightness and expressible moisture. These results describe the changes induced by HPP on sarcoplasmic proteins and confirm a relationship between modification of the sarcoplasmic protein fraction and alteration of meat quality characteristics.     

Antihypertensive effect of a bovine lactoferrin pepsin hydrolysate: Identification of novel active peptides

The potential of bovine lactoferrin (LF) as a source of antihypertensive peptides has been examined. For this purpose, LF pepsin hydrolysate with molecular mass lower than 3 kDa (LFH < 3 kDa) was prepared and orally administered to spontaneously hypertensive rats (SHR), resulting in reduced systolic blood pressure in a significant and maintained manner up to 24 h after administration. LFH < 3 kDa was further fractionated by semi-preparative high performance liquid chromatography (HPLC) and 38 peptides, contained in the active fractions, were identified by using an ion trap mass spectrometer. Based on the peptide abundance, a total of 11 peptides were chemically synthesized and their ACE inhibitory activity tested. Only three of them, corresponding to peptides of sequences LIWKL, RPYL and LNNSRAP exerted in vitro inhibitory effects on angiotensin I converting enzyme (ACE) activity and had a 50% inhibitory concentration (IC₅₀) of 0.47, 56.5 and 105.3 μM, respectively. The three peptides also showed antihypertensive effects in SHR and remarkably the effect of LIWKL remained significant for up to 24 h post-administration, similarly LFH < 3 kDa and the captopril control. The two most potent in vitro inhibitory peptides showed ex vivo inhibitory effect on ACE-dependent vasoconstriction as well. In conclusion, three novel LF-derived peptides and a pepsin LFH < 3 kDa lowered blood pressure and exhibit potential as orally effective antihypertensive compounds.     

Chemical composition and characterisation of seeds from two varieties (pure and hybrid) of Aesculus hippocastanum

Investigations have been conducted on some samples of naturally desiccated horse-chestnuts (Aesculus hippocastanum), representative of the two most common mediterranean varieties: the pure species (AHP, giving white flowers), and a hybrid (AHH, giving pink flowers). Different experimental techniques have been used to gain more information on morphological structure and chemical composition of these complex matrices. Surface analysis by SEM showed no differences in such floured samples (wild type), while thermal behaviour (DSC) outlines some significant differences between them. Chemical composition reveals some differences in residual moisture (AHP = 6.97%; AHH = 6.59%), proteins (AHP = 2.64%; AHH = 1.82%), lipids (AHP = 4.13%; AHH = 5.10%), glucides (AHP = 15.2%; AHH = 14.3%), and ashes (AHP = 2.51%; AHH = 2.19%). Most likely, these characters modulate other undifferentiated chemical parameters, such as cold water solubility (CWS:AHP = 53.9%; AHH = 48.6%), and total inorganic soluble salts (TISS:AHP = 2.18%; AHH = 1.92%). Principal component analysis was applied to differentiate the two horse-chestnuts varieties. In particular, the first principal component effectively distinguish and discriminates AHH and AHP samples in two well-separated categories, giving, at the same time, some information on the influence of the whole set of chemical compositional parameters.     

Effects of intercropping, Bradyrhizobium inoculation and chicken manure fertilisation on the chemical composition and physical characteristics of soybean seed

A field experiment was carried out to investigate the effect of two Bradyrhizobium strains (local and imported), chicken manure fertilisation (7 t/ha) and intercropping with sorghum on the chemical composition and physical characteristics of soybean seed. For both monocropping and intercropping systems, moisture content slightly increased for both systems and for all treatments, while ash, fibre and carbohydrate contents fluctuated for both systems and treatments. The protein content of the seeds was significantly (p ? 0.05) increased for all treatments. Tannin content was increased significantly (p ? 0.05) with a concomitant decrease in protein digestibility for both systems and for all treatments. The seed weight (100 seeds), hydration coefficient and cookability were increased for all treatments. Mineral composition of the seeds was increased and the increment varied with different treatments.