The Influence of silver and zinc oxide Nanoparticles on germination of palmAbstract

In this project, we will investigate the influence of Silver (Ag) and Zinc Oxide (ZnO) Nanoparticles on germination of date palm known as Phoenix Diffusa. The proposal will demonstrate improvement in the palm by using the biosynthesized Ag and ZnO NanoparticlestechniquethanapplyingchemicallysynthesisNanoparticlesforthegerminationprocess.TheNanoparticlesthatwill support the germination are to be synthesized with plant extracts for examples BoerhaaviaDiffusa and PongamiaPinnata. BoerhaaviaDiffusa and PongamiaPinnata are leaf product and they serve as reducing agents for Ag and ZnO Nanoparticles respectively.UsinggreensynthesizedNanoparticleswillhelptominimizethetoxicityeffectthatarisesfromothermethods.Weare toapplyAgandZnONanoparticlesthanotherexistingdifferentNanoparticles;gold,iron,copper,alloy,magneticetc.becausethey can highly protect plant from harmful disease. Silver and zinc oxide are also mostly available and cheaper than gold Nanoparticle. Maximizing the germination of the palm through the influence of biosynthesized Nanoparticles will be reported. The Date palm is atypeofplantthatneedswarmthorheatatitshighqualitytogrowwell.TheuseofmetalNanoparticlesinthisworkwillefficiently generateheat,whichisstronglyenhancedbyPlasmonresonanceinthepresenceofelectromagneticradiation.Transmissionelectron microscope (TEM) and spectrometer are to be used for the characterization of the resultant images’spectra.

Figure 1 shows a flow chart for an overall view of the project settings.

Figure 1: A flow chart of Date palm gaining heat from biosynthesized Ag and ZnO Nanoparticles

Literature Review.

UsingmetalNanomaterialshasincreasedbecauseoftheirnumerousadvantagesandgoodimpactonmanyfieldsoflife,forexample, in agriculture, [1]. The application and properties of Nanoparticles is dependable upon material, shape and size. Hence, the intense exploration by many researchers to produce synthesized Nanoparticles that will be of high benefit to the environment, particularly to their numerous fields of application. The effects of Ag and ZnO Nanoparticles on plants have been under investigations [1-6]. Plantgrowthparameterssuchastheheight,thenumberofleavesandseedshasbeenusedtodeterminetheplantreproductionwhen treatedwiththeZnONanoparticles[6].ZincoxideNanoparticleshaveplausibleattentionbecausetheyareparticularlyantibacterial, antifungal, Ultraviolet filtering features, high catalytic and photochemical activity [7-8]. PongamiaPinnata was used as reducing agent for ZnO Nanoparticles and it contains a wide range of biologically active compounds with functional properties such as anti- inflammatory, antioxidant, antifungal, antimicrobial [9], anti-lipid oxidative, anti-diarrheal and anti-ulcer [10].The Silver Nanoparticles are very successful towards diagnosing and treating cancer [11-12]. For the Ag synthesis, BoerhaaviaDiffusa was the plant extract used as reducing agent. The analysis revealed an average particle size of the Nanometer scale of Silver Nanoparticles having face-centered cubic structure with spherical shape [13]. Reference [14] stated that the application of micro- nutrientfertilizersasNanoparticlesisanessentialpathtoproductionofrequirednutrientsgraduallyandinacontrolledway,thatis useful to mitigate the problems of pollution from fertilizers. It is because of that when there is a transformation of material to a Nanoparticle their morphological, chemical and biological changes tends to facilitates characteristics activities [15]. The Nanoparticles will however need to undergo a green synthesis in order that a minimal toxicity release to agricultural environment. Different processes that involve the biological synthesis of microorganisms, plants, enzymes, and alga have been proposed as feasible environmental friendly alternatives to chemical and physical methods because of the harmful and costly [16-19]. Again, the importance of using natural, renewable and low cost plant extract could be able to produce the metal oxide Nanoparticles with aqueousmediumbyavoidingthepresenceofhazardoussubstanceandtoxicsolvents.Therehavebeendifferenttechniquesreported for the synthesis of Nanoparticles such as light amplification stimulation emitting radiation, photo-chemical reduction, electromagnetic processes and bio-synthetic methods. Plant extract based on biological synthesis undergo highly controlled assembling which makes them suitable and less harmful for metal Nanoparticle applications [20]. Under natural conditions the germination and reproduction of date palm may take several years, usually with a low rate and uneven. Palm is a type of plant that needswarmthorheatwithahighqualityseedforahealthygermination.MetalNanoparticlesefficientlygenerateheatwhichis

(1

)𝑵�

strongly enhanced because of resonating plasma at light radiation [21]. In this work, Ag and ZnO Nanoparticles which have free electrons have been shown with their ability to transfer warmth to the surrounding.

Project Objectives

1. Use of Ag and ZnO Nanoparticles synthesized with plant extract, which is better than high toxic chemical synthesize Nanoparticles

2. SettingupofasupportingwarmthsurroundingforpalmdevelopmentobtainedfromAgandZnONanoparticles3. Characterization ofNanoparticles.4. Analyzation of the influence synthesized Ag and ZnO Nanoparticles on palmgermination.

Theoretical background.

Theoretically,metalNanoparticlesarenotgoodlightemittersthatcanreleaseheatefficientlythroughopticalexcitation.Theoverall heatgenerationwillbecalculatedbyaknownopticallyabsorptiverateformula[21].Mostessentialparametricforheatapplication fromNanoparticlesofNanoscienceisthetemperatureincreaseandasimplemechanismisneeded[22-23].Thismechanisminvolves a strong drive of light amplification by stimulated emission of radiation electrical fields, free carriers in the Nanoparticles, where theheatwillresultfromenergyacquired.ThedistributionalheatroundtheNanoparticlesthatisexcitedbyopticswillbeexpressed with the equation of the heat-transfer asshown

in [21]: 𝜌(𝒓)𝑐(𝒓)𝛿𝑇(𝒓,𝑡)=∇𝑘(𝒓)∇𝑇(𝒓,𝑡)+𝑄(𝒓,𝑡) (1)𝛿𝑡In this project, the local temperature is T( 𝒓, t) while coordinates are r and t timing. The parametric materials’ c(r), ρ(r) and k(r) will be heat-specification, mass-density, and thermal-conduction, respectively. Dissipative lightning that energizes theNanoparticlesisrepresentedasoffunction𝑄(𝒓,𝑡).Maxwell’sequationsareusedtocalculatethefieldsfromasystem.The equation 2 will describe an ordinary heat surrounding a Nanoparticle at steadily situation[21]:

∆𝑇(𝒓) = 𝑽𝑵��

𝟒𝝅𝒌𝒐𝒓 ; ∆𝑻𝒎𝒂𝒙∝ �𝟐, (2)

The distance from the center of a Nanoparticle and the environmental heat conduction is r and Ko, respectively. 𝑽𝑵�is the Nanoparticles volume [21]. Half of the diameter of the Nanoparticles to the secondary degree is proportional to increase in temperature. The surrounding medium of the heated Nanoparticles can be a chemical solution, polymer and water with presents of living things such as plant and organisms. Figure 2, shows the generated heat Q can be calculated analytically based on the assumption that the radius of the Nanoparticles radius is small compare to light of incidence and the wavelength.

Figure 2: (a) Schematic of photonic driven Nanoparticles (b) The temperature increase for Ag and ZnO are functional separations between the environmental medium and middle of Nanoparticle [21].

In the figure 2, dielectrically parameter of the Nanoparticles and environment are represented with ƐNP and Ɛo, respectively where theexplanationisinreference[24].Theheatgenerationrateandincreasingtemperaturesdependsphysicallyontomaterialfeatures. Date palm growing requires dry, medium moisture and hot temperatures above (75º F) to survive. Increasing the heat to higher temperature around 90º F allows seeds to discard its shell, resulting in faster germination.[25]. An indicator of acceleration and speed of the date palm germination are to be calculated based on Mean germination time (MGT) with the following equation[26]:

𝐺𝑇 = � ∑𝐧𝐢 x 𝐝𝐢𝑁 (3)

The days and number of seeds is represented with niand di, respectively where N is the total number of seed germination during the experiment.

The Experimental process:

Inthiswork,differentprocesseswereinvolvedtodevelopthedatepalmsthatgrowthroughtheinfluenceofbiosynthesizedAgand ZnONanoparticles.Firstly,extractsofPongamiaPinnataandBoerhaaviaDiffusawillbepurchasedfromanonlineshop(Alibaba). For biosynthesis of silver Nanoparticles, a mixture of 10.0 mL of BoerhaaviaDiffusa solution and 1.0 mL Silver nitrate aqueous will be prepared. Following by the process of placing the mixture in the magnetic stirrer for 2 min to produce Ag0 (reduction process) which can be obtainable by monitoring the colour changing of the resultant solution[32].Similarly ZnO NPS will be synthesized by adding two millilitre of Zinc acetate dehydrate to 50 ml of PongamiaPinnata extract. Then, the reaction will need 2 hrs to complete with the stirring process at 70 Cº. Next, a white powder will be obtained by centrifugation at 4000 rpm for 10 min. Following by washing process by distilled water and then they will be dried at 100 Cº [33].Reduction process of Ag and ZnO ions will be monitored by UV absorption spectra measurement to approximately 72 hours. UV also will be used for the formation and stability of produced Nanoparticles. A HITACHI, model U-2900/2910 is to be used to record the spectroscopic measurements of UV-Vis from 300 nm -580 nm spectrophotometer. The ZnO and Ag Nanoparticles is expected to disperse in deionization at 374 and 420 nm, respectively[27,28].

Figure 3: Absorptive spectra of ultraviolet visible for (a) Silver (b) Zinc oxide under the synthesis of extracts BoerhaaviaDiffusa and PongamiaPinnata respectively [27]. The insets are the TEM image of Ag and ZnO Nanoparticles [28].

Insetsoffigure3shows,thebiologicallysynthesisofZnOandAgNanoparticlesaretobedry-freezeandkeepinpowderformfor material characterization using Transmission electron microscopy (TEM). The absorptive spectra of ultraviolet visible from BoerhaaviaDiffusaandPongamiaPinnataextractsreactingatvariousintervalsisthegraphoffigure3(aandb)withtheexhibition ofAgandZnONanoparticles.Theresonancepeaksasshowninthefigurebetween420and425nmwhichisprominentwillconfirm the formation of the Nanoparticles. In addition, energy dispersive x-ray spectroscopy(EDXS) and Field Emission SEM (Scanning Electron Microscope) will be used to examine the shape and size ofNanoparticles[27,28].

Figure 4: (a) Set-up for the experiment which will have ice in a container of Ag and ZnO Nanoparticles. Inset of Ag and ZnO Nanoparticles with the light excitation and the transformation of warmth to the icy surrounding medium [21]. (b) Analysis of variance examine the different between the expectation from palm treatment with Nanoparticles and without treatment palm.

Secondly, the required temperature is to be generated from the Nanoparticles through experiment conduction shown in figure 4a which include the usage of thermocouple meter (4mm) and glass substrate of silicon (0.2,0.13 mm) that will be purchased from Cole-Parmer (online shop). The heat will be generated by optically driven synthesized ZnO and Ag Nanoparticles and will be characterized based on the experimental setup of figure 4a. Recording is by placing the ZnO and Ag Nanoparticles solution (20µl) for the medium of established thermal dynamic features (icing)(-20Cº) and recording change in temperature surrounding the Nanoparticles after exciting optically. The amount of heat that will be generated from the optically excitation of ZnO and Ag will becalculatedfromdevelopedtheoryoncethereisachangeinlocaltemperature.Theexperimentalapparatuswillbeusedtorecord change in temperature surrounding the Nanoparticles during optical excitation (laser intensity will be monitored between 0.2 and 50 mW. Locating and numbering the Nanoparticles in the icing will be determined as some light intensity thresholds that cause meltingforspecifictemperatureprobing.TherewillbecalculationsofincreasingtemperatureonthetopofZnOandAgondifferent sizes. Temperature increase can become noticeable if the Nanoparticles size is sufficiently large. The heat generated by these Nanoparticles optically will create a phase transforming to the surroundings and such processes are explained in the equation1.

Finally, the phase transformation of heat generation from the surface of ZnO and Ag to surrounding of date palm will improve the germination as analyzed statically. For the germination experiment as reported in the reference [30]. Firstly, the Date palm seeds(Medjool)(30seeds)of13mmaretobeobtainedfromAlibaba(onlineshop)andtheywillbewashedbydistilledwater[31]. After that, half of seeds will be soaked in each Nanoparticles solution of 20% and 60% and the remaining seeds will be untreated. Subsequently, the mean germination time will be calculated using the equation (3) and an additional calculation of germination present(GP=𝑛𝑖/�×100),thegerminationrate(GR=∑𝑛𝑖/𝑡)andvigorindex(Vi=GP×(rl+sl)whererlandslrepresent the root and shoot length respectively. The data from germination rate, duration, the percentage, shoot, root and weights are to be analyzed using SPSS version 22.0. Analysis of variance (ANOVA) will be used to examine the different between the palm treated with Nanoparticles and the untreated palm. Figure 4b shows that by using biosynthesized ZnO and Ag that there is potentiality of palm germination enhancement. A warmth humid surrounding released from photonic driven Ag and ZnO Nanoparticles will provide a healthy palm which requires temperatures above 75º F to survive. Though, a date palm may tolerate extensive dry condition at very warmth environment, high water is needed for healthy growth and reproduction [29] which is the key idea of theproposal.

The Overall Impact

The technique that will be employed in the development process of the date palm for this project will assure an eco-friendly environment. The project aim is to use green synthesized Nanoparticles to support the growth of the palm. Moreover, the reason behindtheuseofAgandZnOamongotherNanoparticlesisthattheyarecheapandhighlyavailable.SaudiArabiacanbecurrently considered to be the homeland of the date palm because it is one of the most important national products. In 2014 German-Saudi BusinessMagazine(GSBM),reportedthatmorethan20millionvarietiesofthedatepalmwithdifferentqualitiescoversmorethan 150,000 hectares in a Saudi Kingdom and this generates billions of USD to the country annually. The GSBM also estimated palm production of 1.1 million tons which accounts for approximately 72% of the country’s total agriculture each year. This project can significantly boost the economy of the country by more than 20% because of the provision of engineered warmth condition need forfasterpalmdevelopmentasdesiredbysomeregioninSaudiandothercountriesofverylowaveragetemperature.

Project Team: The project will involve a team of three researchers and their job responsibilities during the period are stated below:

1. Professor Losa is the principal investigator for this project. Prof Eid has an advanced experience in Nanoscience with a very good research reputation in Sudan University over the years. She is originally from Saudi with good knowledge of important date palm to the country. She will direct, examine and present results toconferences.

2. A research associate Dr James O’Shea is an expert working with different optical, electronic and mechanical equipment in a complexscientificlaboratory.Heistosetupthelabfortheexperiment,train,supportandmonitorthePhDstudentashecarriesout the work. O’Shea will help write some paper forpublication.

3. A PhD student who specializes in agricultural science. He will review related articles, monitor the palm germination, synthesize Ag and ZnO Nanoparticles, then collect data and characterizing the different material. Finally, he will write the required scientific reports or thesis based on theresults.

Finance management:

Firstly, the flowing table highlights the potential cost (£) of individuals participating in the project including the effort and percentage of work for each researcher.

Grade Name Appointmentdate

Duration of appointment

Full time

%

Salary Pension .Nat. Ins Total Total coston

grant(£)SRS08 Losa 10/01/2017 36 100

RS07 JamesO'Shea

10/01/2017 36 100 29541 2,203 4727 36,471 112,791

PhD Will Ruth

10/01/2017 41,160

TS05 Helen Davies

10/01/2017 36 50 22057 3529 1560 27146 42009

The total cost: £ 195960

Secondly, the table below shows the detailed cost that will be allocated to consumable and equipment.

Consumable Cost (£)

Equipment Cost (£)

Chemical component such as AgNo3, distilled water and Zinc acetate dehydrate.

670 Magnetic stirrer, UK (Stuart Analog Stirrer, 12" x 12", stainless steel top plate, 120 V)

823

Date palm seeds (Medjool) and extracts of Boerhaavia. Diffusa and PongamiaPinnata

290 Ultrasonic cleaner, UK (Cole-Parmer long-bed Ultrasonic cleaner, 220 VAC)

2353

Electric usage. 3680 Ultraviolet spectrometer UV ( UK,Jenway 63 Series Visible.Spectrophotometer;115VAC)

1965

Computer system. 3500 Thermocouple meter (Digi-Sense Calibrated Water- Resistant Thermocouple) UK Thermometer.

154

Page charges for journals. 2300 TEM, EDXS and FESEM In lab

IP protection. 13500 Total 5295Glass wafer (0.2,0.13 mm). 400Other laboratory work and materials. 1000Total 25340

3- Facilities.

Access for the university photo physics labs for imaging and sensing by using various microscopese.g. TEM and FESEM.

£5650

Access for the university chemical lab for biosynthesis of Nanoparticles £3360Access for the university science environmental labs for setting the germination experiment. £2800Total £11810

4- Travel andsubsistence.

Distance Details Cost (£)

47thInternational Conference on Environmental System July 16-20th,2017 ~ Charleston,SC 320010thInternational Conference on Nanotechnology Fundamentals and applications (ICNFA’17) December

15th, 2017, Canada2500

Total 5700

The total project cost will be £ 342085 for a three-year project (156weeks) distributed in the table below

Staff £ 195960Consumable £ 25340Facilities £ 11810Travel and Subsistence. £ 5700Equipment. £ 5295Sub-total £ 244105Indirect cost £ 97980

Timeline: The attached Gannt chart stated all the work that will be carried out for a period of three years. The project is proposed tostartearlyJanuary2017andendtowardsthelastweekof2019.Thelongerperiodasassignedtosometitleincludestheholidays and break that will come up during theproject.

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