Evaluation and Use of Nem (Azadirachta indica A. Jus) as Organic Substrate Componet by Cody Wyat Kiefr A thesi submited to the Graduate Faculty of Auburn Universiy in partial fulfilment of the requientsor the Dgree of Mtr of Scinc Auburn, Alabam ay 9, 2011 Keywords: urease, potting soils, commercial media, compost, poultry protein compost Copyright 2011 by Cody Wyat Kiefr Approved by Jef L. Sibley, Chair, Profesor of Horticulture Charls H. Gilam, P Glnn B. Fan, Astnt Profsor ofortiult H. Alen Torbert, Reerch Director of USDA Soil Dynamics Lab ii Abstrac With ?green-mindednes? increasing n popularity, the use of organic substraes has become even more iportant in the retil horticultural market. Multiple experimnts were conductd to evaluate commercialy available bagged substraes. Th efcts of lven commercialy ilabl substras on plnt growth were evaluatd using four species of plnts. In experiment one, Jungle Growth ? organic substraes outperformed other commercial potting substras and wre only slightly inferior in perfance to the best-ranking substrae, Miracle-Gro ? Moisture Control ? . The scond experimnt revealed that, where difrent, the Jungle Growth substraes outperformed al other substras tted (though not staistcly greater than Sta-Gren ? Flower and Vegetable mix when usd on tomatoes and petunis). The srch for benefical mndmnts for horticultural soiles mdi is a constant and ongoing proces. Ne (Azadirachta indica A. Juss) cake powder is currently being used as a substra component, but its full efets on the rhizosphere and subsequent plant bioms have yet to be explored. Therefore, a third study was conductd to evaluate nem at varying percentages ranging from 0-5% in two stock substraes ? one containing poultry protein compost and the otr containing peat in place of the compost and receiving nutrints via fertigaton. Result show the benefit of t addition of neem, espeialy at 5%, in the compost-containing treatments. The same cnnot be said of the non-compost treatment. Plants grown in 0-1% ne iii treaments within the non-compost mixes had result that outperformed any of the compost treatents and often outperformed other non-compost treatmnts. Trefore, neem appers to benefical when amnded into poultry protein compost-containing substras, but antagonistin added to standard mixes that wil be fertigated. Nursery and greenhouse growers continue to sek mterils to decrease costs of plant production whil maintning environmental stwardship. Incorporation of nem cke as substrae component could potentialy impact nirogen releas a result of altering stra baterial activiy. A prelinary study investigad the impact of nem on substrae gas rels nd provides a starting point forurther investigaon regarding nee us as substrae component. With three substrae groups being testd with varying percentages of nem, this study reports on both across-group result as wl as within-group result. Across al three groups, 3% neem within the pine bark + poultry protein compost + neem group was significantly greatr in CO 2 production than al treatmnts within the pine bark + neem group as wel as zero percent nem within its own group and the pine bark + poultry protein compost + 19-612 + ne group. Nitrous oxide emison was significantly greater in the pine bark + poultry protein compost + 19-612 + neem group than al other tretmnts. Within-group comparisons reveal that three percnt ne had greater CO 2 eison than zero percent nem for both the pine bark + nem and pine bark + poultry protein compost + nem groups. Three percnt neem also produced significantly greatr CH 4 than zero percent nem in the pine bark + poultry protin compost + neem group ? and within the sa substrae group, two percent nem had significantly greatr N 2 O emison than zero percnt nem. There wre no significant iv diferencs among treatents within the pine bark + poultry protein compost + 19-612 + ne group for al thre gas analyzed. v Acknowledgements The author, first, foremost and most iportantly, would like to thank God, ?Who delivers me through Jesus Christ our Lord!? (Romns 7:25 NIV). It is only by His grace tha the author has drawn breath and strength to acomplish the tasks set before him. The author?s life was bought by the spotles blood of the Lamb and t author is fver indebtd to this supreme act of grac and love ? his life and soul are Yours, Lord! The author ows a great mesure of thanks and love to his parents, who walked with him through the entire university experienc with support and guidance ? thanks Mom and Dad! Along t same lines, the author would also like to thank his four-legged friend, Oto, who never csd to bri with excitemnt at spending time with hi and who strode with him through many trials. Drs. Sibley and Gil, much thanks to you for taking a chance on me. With only a question of intrest from the author, you did not stop until he walked through the doors of Funches a masters student. Thank you Drs. Fain, Torbert and Wats, for your help throughout the resrch and thesi proces. hank you Drs. Ponder and Eakes for alowing me to work beside you and gain valuabl xperienc taching. As for the rest of you ?hortis?, thanks for your support and friendship. Mt, Jeremy and Chris, you took a lot of time out of your schedules and lives to guide the author through the research proces, so thanks. vi And Jes, what can the author say? The Lord has been so faithful in bringing you in his lif and in giving you t patienc to put up with him. You have been invaluable, not only in your help with resarch, but also a his bet friend. As the author writs thi, the countdown to marrige begins, but as you read it, the liflong journey of serving God togetr wil have alredy begun. The author loves you and thanks you for everything! Onward you, Oto and the author go togetr as a family! There is only one thing left to say: Prais God from Whom all besings flow!! vii Table of Contents Abstrac...............................................................................................................................ii knowledgmnts................................................................................................................v List of Tabls...................................................................................................................i ist of Abbreviations.........................................................................................................ix Literature Review.................................................................................................................1 Evaluation of Nm Cake Powder Percntages an Organic Substrae Component......3 Gas Production from Soiles Mdia Amnded with Nem Cake Powder........................30 Final Discusion.................................................................................................................50 Appendix A: Commercialy Available Soiles Mdia Comparisons................................53 vii List of Tables Table 1. Growth indices, pH, EC and SPAD 28 DP for salvi from neem percentages study 2010..........................................................................................................................28 able 2. Growth indices, pH, EC and SP 56 DP for salvi from nee percentages study 2010..............................................................................................................29 Table 3. Ga analysis reults for organic urease inhibition substrae amndment.............46 bl 4. Pine bark + neem within-group gas analysis reults............................................47 Table 5. Pi bark + compost + neem within-group gas analysis reults..........................48 bl 6. Pine bark + ct + ne + 19-612 within-group gas analysis reults.........49 Table 7. Growth indices, pH, EC and SPAD on petunia in a 2009 commercial bagged subsrae sudy........................................................................................................69 able 8. Growth indices, pH, EC and SP on marigold in a 2009 cercial bagged subsrae sudy........................................................................................................70 Table 9. Growth indices, pH, EC and SPAD on dianthus in a 2010 commercial bagged subsrae sudy........................................................................................................71 able 10. Growth indices, pH, EC and SP on marigold in a 2010 cercial bagged subsrae sudy........................................................................................................72 Table 11. Growth indices, pH, EC and SPAD on petunia in a 2010 commercial bagged subsrae sudy........................................................................................................73 able 12. Growth indices, pH, EC and SP on tomato in a 2010 cercial bagged subsrae sudy........................................................................................................74 ix List of Abbreviations EC Electrial Conductiviy GI Growth Index PB Pine Bark PC oultry Protein Compost JG Jungle Growth ? MG Miracl-Gro 1 Chapter I Literatue Rview Soiles Mdia: Beginning in the 1970?s, pine bark quickly became an invaluable resource to the horticulture industry. Present-day demands for this comodity, howver, have also increased and, with the growth of the horticulture sector, our industry is feling the presure to find some altrnatives to exclusive pine bark usage (Avent, 2003). While greenhouse plants arelmost exclusily produced in pet-substraes, 75-100%, by volum, of continer substraes in the eastern US are comprisd of pine bark (Lu et al., 2006). Future projections projct t rising cost of pine bark cbid with les availbiity to the hortiulture industry (Lu et al., 2006). Numerous organic and inorganic substances are being used to develop varying substras. Clan chip residual (the mteril lft on the forest floor following timber harvest) cn be a relistc alternative to pine bark-exclusive substraes for ornaental plant production (Boyer et al., 2008). Fain et al. (2008) succesfully usd WholTre (whole pine tree shoots) in varying percentges a suitabl grenhouse mdia for growing marigolds and petunias. With additional frtilzer, Jckson and Wright (2007) were able to grow plants in 100% pulverized pine wood. Cotton gin compost was usd as a viabl substrae component by Col t al. (2002). Spent tea grinds can be succesfully used in 2 horticultural substraes (Wls, 2008). Poultry liter was a suitable amndment for substraes omprisd primarily of either pine bark, WholeTre (cretd by chipping, then further milng entire pine trees ? log, limbs, needls and bark), or clean chip residual for growing plants in contairs with favorabl result and is a readily availble source of nitrogen (Fin et al., 2008; Mrble et al., 2010; Mitchel and Donald, 1995). Parboiled fresh rice hulls can be used sucesfully as a substrae component comparabl to perlit (Evans and Gchukia, 2007). Using stras from peanut hulls, pine bark, peatmoss and combination thereof, Bilderback et al. (1982) reported succes in growing azles. In searching for an ecologicaly-friendly alternative to rockwool, Urrestarau et al. (2005) concluded that almond shels can be an efcti and benefical soiles mdia for plant production. Coconut coir c a vible substrae for plnt production, however its succes depends on particle siz (Noguera et al., 2003). Peat- vermicult substras rved as viable substras for growing chrysanthemums (Pul and Le, 1976). Mineral substraes have the potential to perform silrly to peat-based mixes (Sith e al., 1995). Smith and Hal (1994) determined that a perlite-sd substrae can be comparable in manageent and productiviy to peat-basd potting mixes. While potntial aternative substraes sm to abound, there a requirements that must be mt of the mtril in order for it to be usable and efective. Chang and Lin (2007) report that basic requirements for a succsful and benefial plnt-growing medium include: excelnt cheical resitance properties, light weight, inexpensivenes, abs of pest and disae and availbiity Nkongolo and Caron (2006) noted that particle siz, specifly in pet and pine bark-based substraes, influencs plant 3 response. Handreck (1983) reports the importance of particle siz and advise that substra formultors heed the ?fines? fraction of a substra, especialy particl size smaler than 0.5mm, as thi size controls the physical properties of pine bark-basd substras. Shrinkage is a physical property of organi soils (such as peat) that must be managed (Schw?rzel et al., 2002). Nemati e al. (2002) noted that insufficent aration of rtifcl growing mdia is a comon proble in greenhous production. Neem: The soiles mdia amendmnt in question is a product of Azadirachta indica A. Juss, or t nem tre. The ne tree is known as the ?vilage dispensary? in Indi and Southeast Aia, where its native (Biswa et al., 2002). This i due to the result of both scienc and tradition about the medicinal and agricultural uses that neem provides. The nem tre is an evergreen tre belonging to the mahogany faily, Mliac, of which Melia azedarach Chinaberry is alo a member (Bisw et al., 2002). Steped in Indian tradition and lore, the neem tree continues to play roles in their traditions, mdicine and agriculture and now t rest of the world is beginning to pay more atention to neem, whih has been somewhat controversial to the traditionalist in Asia (Mrden, 1999). Almost every part of the neem tree (bark, leaves, seds, fruit, flowers and roots) i used in various ways, with more than 140 cheicl compounds having ben isolated by scintst (Brahmchari, 2004). These compounds have proven efective against and to ombat intestinal worms, treat ashma and rheumatism, a wel as being useful as analgesic, anti-malril, anti-fungal, insectidal as wel as ?general health promoter? (Brahmahari, 2004). Al of these uss alo sem to com with litle to no il efct to humns (Mrden, 1999). 4 The cmicals iolated from neem can be categorized into two groups: isoprenoids and non-isoprenoids (Brahmchari, 2004). Non-isoprenoids aremino acids, carbohydrates, flavonoids and others, while isoprenoids contain compounds such as zdirachtin (Brahmchari, 2004). This lat compound, azdirachtin is used today in many insetides in the United Staes. In the early 1990?s a company in Florida patentd stbilizd version of azadirachtin to increse it usefulnes and was then approved for use on food crops by the Environmental Protction Agency in 1994 ? t first neem- basd product marketed in t U.S. (Mrden, 1999). Many studies have followd in search for the insctidal and medicinal applictons for compounds including neem, but other uses for neem have not ben resrched in the United Staes. Neem has ben used as a soil amendmnt in many studis in India. Neem sed residue provided a nitrogen value, aftr oil extraction, of 7% and at a reles rat fast enough to satisfy maize nutrition (Agbenin et al., 1999). Neem oil aso nhanced plnt growth when incorporatd into soil at 2.5, 5.0 and 20 ppm, though 10 ppm decreasd plant growth (Bhaskar and Charyulu, 2005). Likewise, Agyarko et al. (2006) reported tha soil nutritional levels increased with poultry manure and increasing levels of nem leaves. Bhala and Dvi Prasd (2008) reported higher vegetive growth in plants than reproductive growth (both showing higher growth than the control) when neem cke was incorporated into the soil. Nm?s ipact wihin a substrae has been atributed to its potential urese retardation activiy (B. Hurst, personal communication). Ureas i the naturaly occurring enzyme in soil responsible for the hydrolysis of ure into carbon dioxide and amonia (Mnunza et al., 1997). Amonium and amonia are the nitrogen sources 5 within fertilzrs (be it synthetic fertilzr or organic), but the amonia cn undergo volatiltion, which may dereas nitrogen available to t plant. Therefore, it is often desirable to have certin levels of urese inhibition in order to constrain amonium production, resulting in a more readily available supply of nitrogen over a longer period of time. The aasing of amonium cn ld to problems other than amonia volatilztion, such as nitrie toxicity and damage to young plants (Brener and Douglas, 1971). Urease prevalenc in soil is closely linked with organic mater content within a soil or substra (Burns et al., 1972). Fishbein et al. (1973) have shown that urease enzyme purified from jack bean mel may actualy have more than a dozen molculr forms. Each dimr of urese i composd of two half-units, which mans that the enzyme itslf cn be disociatd into constiuents (Fihbein et al., 1973), though it is not stad whetr these subunits themselve are functional. However, if neem breaks down the enzyme dimrs, it is currently unknown if the urease then becoms non-functional. There have been some directed studies aimd at ures and/or nitrificaton inhibition. Burns et al. (1972) showd tha pronase, a proteolytic enzyme, degrades jack bean urease, but has no efect on urease within the soil matrix. Bremner and Dougls (1971) found that metali slts ontinng silver, mercury, gold or copper can have an efct on ureas activiy (silver sulfate had a 48% inhibitory efect). Organic compounds such as phenylmercuri aceta and ctchol ibited ureas by 67% and 74%, respectively. Other inhibitors of urease include phosphorodimides, phosphorotriamides and hydroxamic aid (Manunz et al., 1997). In corn production, Schlegel et al. (1986) noted the following urese inhibitors: hydroxamates, heterocycli sulfur compounds, 6 xanthates, organophosphorus insectides, quinones and phosphoroamides, noting phenylphosphorodiamide as the most efective inhibitor. Schlegel et al. (1986) showed tha urease inhibitors did not increas yild of corn but inhibitors had the greates potentil efct when added to surfce applied urea that was not watered in. Efcts of pesticdes on ureases the focus of Ingram et al. (2005) who reportd tha pestides an have a noticbl efct on bacterial urese, but no significant efect on free urease in the soil. Treatment of soils with toluene resultd in increse in soil ureas activiy, as did chloroform fumigaton (Klose and Tabati, 1999). Also, the addition of glucose, or any organic mater that is hospitabl to microbial activiy, was shown to increas ureas function in soils (Zntua and Bremner, 1976). Mohanty et al., (2007) reported on the potential inhibitory efects of neem sd kernel powder on urease in thre mineral soils native to India, showing slight suppresion of urease activiy when applid to acidi soils. M?ndez-Bautist e al., (2009) studied the efcts of nem leaf extracts on greenhouse gas emisons and inorganic nitrogen in urea- amnded soil and reported that the laf extract had no significant efet on urease, but y lit nirificaton. Majumdar et al., (2000) coated ure with nem sd powder before adding to rie filds in North India, resulting in slight nitrificaton inhibition. Kumar et al., (2007) usd neem oils to coat urea and added it to sndy-loam soils resulting in some nitrificaton inhibition as wel. Bhala and Devi Prasad (2008) showed, in one of their studis, that addition of nem cake into a mineral soil is an economical and efctive method for reducing fertilzr applicton by prolonging fertilzr availble to the plant. 7 The question now is whetr neem wil have any efect on urease activiy within soiles potting media, especialy a pine bark-based substra. Sondly, would neem cake us be cost-fctive? Ptra and Chand (2009) noted that though nee is being proved as an efeti soil amendmnt, it is not being usd wide-scal becaus of, in their case, the coating procs for the urea is cumbersome and the mterils are not readily vailbl. Trefore, one area of reserch to be addresd in this theis i to evaluate neem cake?s efectivenes in soils mdia and, if eective, can neem be used in cost- fctive aounts incorporated in the substraes with benefil result to the plant. 8 Literatue Cited: Agbenin, J.O., E.B. Agbaji, I. Suleiman, and A.S. Agbaji. 1999. Asemnt of nitrogen mineralizaton potential and availbiity from neem sd residue in a savanna soil. Biol. Fertil. Soils 29:408-412. Agyarko, K., P.K. Kwakye, M. Bonsu, and B.A. Osei. 2006. Impact of applicaton of neem laves and poultry manure on nutrint dynamics of a Hapli Arisol. Archis of Agronomy and Soil Scienc 52:687-695. vent, T. 2003. So You Want to Start a Nursry. Timber Pres, Inc. Bhala, R.S. and K.V. Devi Prasd. 2008. Ne cake-urea mixed applicatons increase growth in paddy. Current Scienc 94:1066-1070. Bhaskar, K.V. and P.B.B.N. Charyulu. 2005. Efect of environmental fctors on nitrifying bacteria isolted from the rhizosphere of Setaria itali (L.) Beauv. African J. of Biotchnology 4:1145-1146. Bilderbak, T.E., W.C. Fonteno, and D.R. Johnson. 1982. Physical properties of media composed of peanut hulls, pine bark, and peatmoss and their efcts on azl growth. J. Amer. Soc. Hort. Sci. 107:522-525. Biswa, K., I. Chatopadhyay, R.K. Banerje, and U. Bandyopadhyay. 2002. Biological ctivies and medicinal propertis of nem (Azadirachta indica). Current Sinc 82:1336-1345. Boyer, C.R., G.B. Fain, C.H. Gilam, T.V. Galgher, H.A. Torbert, and J.L. Sibley. 2008. Clen Chip Residual: A substrae component for growing annuals. HortTchnology 18:423-432. 9 Brahmachari, G. 2004. Neem?An Omnipotent Plant: A Retrospective. ChemBioChe 5:408-421. Brener, J.M. and L.A. Douglas. 1971. Inhibition of urease activiy in soils. Soil Biol. Biochem. 3:297-307. Burns, R.G., A.H. Pukite, and A.D. McLaren. 1972. Concerning the location and persistenc of soil urease. Soil Si. Soc. Amr. Proc. 36:308-311. Chang, C.P. and S.M. Lin. 2007. The formation and growing properties of poly (ethylene trephthalate) fiber growing media after thermo-oxidative treatment. Matrils Scienc and Engineering 457:127-131. Cole, D.M., J.L. Sibly, E.K. Blythe, D.J. Eakes, and K.M. Tilt. 2002. Evaluation of cotton gin compost as horticultural substrae. SNA Research Conferenc 47:274-277. Evans, M.R. and M.M. Gachukia. 2007. Physical properties of sphagnum peat-based root substraes amnded with perlite or parboild fresh rice hulls. HortTechnology 17:312-315. Fain, G.B., C.H. Gilam, J.L. Sibley, and C.R. Boyer. 2008. Establishment of greenhouse-grown Tagets patula and Petunia xhybrida in ?WholTr? substras. Acta Hort. 782:387-393. Fishbein, W.N., K. Ngarajn, and W. Scurzi. 1973. Urease ctalysis and structure. The J. of Biological Chemistry 248:7870-7877. Handreck, K.A. 1983. Prticle siz and the physical properties of growing media for ontainers. Commun. In Soil Sci. Plant Anal. 14:209-222. 10 Ingram, C.W., M.S. Coyne, and D.W. Wilams. 2005. Efects of commercial dizinon and iidacloprid on microbial urese ctiviy in soil and sod. J. Environ. Qual. 34:1573-1580. Jackson, B.E. and R.D. Wright. 2007. Pine tree substrae: Frtilty requirements. SNA Research Conferenc 52:523-526. Klose, S. and M.A. Tabati. 1999. Urease activiy of microbial biomas in soils. Soil Biol. ochem. 31:205-211. Kumar, R., C. Devakumar, V. Sharma, G. Kakkar, D. Kumar, and P. Panneerslvam. 2007. Influenc of physiochemicl paraetrs of nem (Azadirachta indica A. Juss) oil on nitrificaton inhibition in soil. J. Agric. Food Che. 55:1389-1393. Lu, W., J.L. Sibley, C.H. Gilm, J.S. Bannon, and Y. Zhang. 2006. Estimation of U.S. bark gneration and iplicatons for horticultural industries. J. Environ. Hort. 24:29-34. Majumdar, D., S. Kumar, H. Pathak, M.C. Jain, nd U. Kumar. 2000. Reducing nitrous oxide emison from an irrigated ric field of north Indi with nitrificaton inhibitors. Agriculture, Ecosystms and Environment 81:163-169. Manunz, B., S. Deiana, M. Pintore, V. Solinas, and C. Gsa. 1997. A molecular modeling study of the urease active site. J of Moleculr Structure (Theocm) 419:33-36. Marble, S.C., C.H. Gilam, J.L. Sibley, G.B. Fain, H.A. Torbert, T.V. Galgher, and J.W. Olive. 2010. Evaluation of composted poultry liter as a substrae amendmnt for WholeTre, Clean Chip Residual, and pinebark for container grown woody nursry crops. J. Environ. Hort. 28:107-116. 11 Marden, E. 1999. The Nem Tree Patnt: International Conflict over the Commodificaton of Life. Boston Collge International and Comparative Law Review 22:279-295. M?ndez-Bautista, J., F. Fern?ndez-Luque?o, F. L?pez-Valdez, R. Mendoza-Cristino, J.A. ontes-Molina, F.A. Gutierre-Miceli, and L. Dendooven. 2009. Efect of pest crolling nem (Azadirachta Indica A. Juss) and mat-raton (Glirdia spium Jaquin) laf extracts on emison of greenhouse gas and inorganic-N content in ure-mended soil. Cheosphere 76:293-299. Mitchel, C.C. and J.O. Donald. 1995. The value and use of poultry manures a fertilzr. Alabam Cooperative Extension System. Noveber 1995. Retrieved from: http:/w.aces.edu/pubs/docs/ANR-0244/ Mohanty, S., A.K. Patra, and P.K. Chhonkar. 2007. Nem (Azadirachta indica) seed kernel powder retrds urease and nitrificaton activies in diferent soils at contrasting moisture and tmperature regims. Bioresourc Tchnology 99:894- 899. Nemati, M.R., J. Caron, O. Banton, and P. Tardif. 2002. Detrmining air entry value in pet substraes. Soil Sci. Soc. Am. J. 66:367-373. Nkongolo, N.V. and J. Caron. 2006. Pore space organization and plant response in peat substraes: I. Prunus xcistena and Spiraea japonic. Sientfic Resarch and Esay 1:77-89. Noguera, P., M. Abad, R. Puchades, A. Maquieira, and V. Noguera. 2003. Influence of particle siz on physical and chemicl properties of coconut coir dust as container mdium. Comm. in Soil Si. and Plant Analysis 34:593-605. 12 Patra, D.D. and S. Chand. 2009. Natural nitrificaton inhibitors for augmenting nitrogen use eficency in soil-plnt system. UC Dvis: The Procedings of the Intrnational Pant Nutrition Colloquium XVI. Retrived from: http:/w.escholrship.org/uc/item/4h30z8tg Paul, J.L. and C.I. Le. 1976. Relation betwn growth of chrysanthemums and aeration of various container medi. J. Amer. Soc. Hort. Si. 101:500-503. Schlegel, A.J., D.W. Nelson, and L.E. Somrs. 1986. Field evaluation of urease inhibitors for corn production. Agronomy J. 78:1007-1012. Schw?rzel, K., M. Renger, R. Sauerbrey, and G. Wesolk. 2002. Soil physical haractristic of pet soils. J. Plant Nutr. Soil Sci. 165:479-486. Smith, C.A. and D.A. Hal. 1994. The development of perlite as potting substrae for ornaental plnts. Act Horticultura 361:159-166. Smith, C.A. P. McDonald, and A.M. Sword. 1995. Culture of hardy woody ornaentals in pure-mineral container substraes. Acta Horticulturae 401:161- 167. Urrestarazu, M., G.A. Mart?nez, and M.C. Sals. 2005. Almond shel waste: possible locl rockwool substiute in soiles crop culture. Scienta Horticultura 103:453- 460. Wels, D.E. 2008. Evaluation of spent ta grinds asn alternative horticultural substrae component. Thesi. Auburn University. Zantua, M.I. and J.M. Bremner. 1976. Production and persistenc of urease activiy in soil. Soil Biol. Bioche. 8:369-374. 13 Chapter II Evaluation of Nem Cake Powder Pcentages as an Organic Substrate Component Signficane to Industry: The sarch for benefical amendmnts for horticultural soiles mdia is a constant proces. Nem (Azadirachta indi A. Juss) i currently being usd as a substrae component, but its full efects on the rhizosphere and subsequent plnt biomas have yet to be explored. Therefore, this study was conducted to evaluat neem cke powder (referred to as neem) at varying percentges ranging from zero-tofive percent in two stock substras ? one containing poultry protein compost and the otr containing peat in place of the compost and receiving nutrints via frtigaton. Result show the benefit of the addition of neem, espeialy at five percent, in the compost-containing treatments. T same cnnot be said of the non-compost treatment. Plants grown in zero-toone percnt ne tretments within t-t mixes had result that outperformed any of the compost treatnts and often outperformed other non-compost treatments. Trefore, neem appers to benefical when amnded into poultry protin compost- containing substras, but antagonistin added to standard mixes that wil be fertigated. 14 Introduction: Beginning in the 1970?s, pine bark quickly became an invaluable resource to the horticulture industry. Present-day demands for this comodity, howver, have also increased and, with the growth of the horticulture sector, our industry is feling the presure to find some altrnatives to exclusive pine bark usage (Avent, 2003). While greenhouse plants arelmost exclusily produced in pet-substraes, 75-100%, by volum, of continer substraes in the eastern US are comprisd of pine bark (Lu et al., 2006). Future projections projct t rising cost of pine bark cbid with les availbiity to the hortiulture industry (Lu et al., 2006). Numerous organic and inorganic substances are being used to develop varying substras. Clan chip residual (the materil lft on the forest floor following timber harvest) cn be a relistc alternative to pine bark-exclusive substraes for ornaental plant production (Boyer et al., 2008). Fain et al. (2008) succesfully usd WholTre (whole pine tree shoots) in varying percentges a suitabl grenhouse mdia for growing marigolds and petunias. With additional frtilzer, Jckson and Wright (2007) were able to grow plants in 100% pulverized pine wood. Cotton gin compost was usd as a viabl substrae component by Col t al. (2002). Spent tea grinds can be succesfully used in horticultural substraes (Wls, 2008). Poultry litr ws a suitbl amndmnt for substraes omprisd primarily of either pine bark, WholeTre (created by chipping, then further milng entire pine trees ? log, limbs, needls and bark), or cln chip residual for growing plants in contairs with favorabl result and is a readily available source of nitrogen (Fin et al., 2008; Mrble et al., 2010; Mitchel and Donald, 1995). 15 Parboiled fresh rice hulls can be used succesfully as a substrae component comparabl to perlit (Evans and Gchukia, 2007). Using stras from peanut hulls, pine bark, peatmoss and combination thereof, Bilderback et al. (1982) reported succes in growing azles. In searching for an ecologicaly-friendly alternative to rockwool, Urrestarau et al. (2005) concluded that almond shels can be an efcti and benefical soiles mdia for plant production. Coconut coir c a vible substrae for plnt production, however its succes depends on particle siz (Noguera et al., 2003). Peat- vermicult substras srved as viable substras for growing chrysanthemums (Pul and Le, 1976). Mineral substraes have the potential to perform silrly to peat-based mixes (Sith e al., 1995). Smith and Hal (1994) determined that a perlite-sd substrae can be comparable in manageent and productiviy to peat-basd potting mixes. While potntial aternative substraes sm to abound, there a requirements that must be mt of the mtril in order for it to be usable and efective. Chang and Lin (2007) report that basic requirements for a succsful and benefial plnt-growing medium include: excelnt cheical resitance properties, light weight, inexpensivenes, abse of pest and disae and availbiity Nkongolo and Caron (2006) noted that particl size, specifly in pet and pine bark-based substraes, influencs plant respons. Handrek (1983) reports the importanc of particl siz and advise that substrae formultors heed the ?fines? fraction of a substrae, especialy particl size smalr than 0.5mm, as thi size controls the physical propertis of pine bark-basd substraes. Shrinkage is a physical property of organi soils (such as peat) that must be 16 managed (Schw?rzel et al., 2002). Nemati e al. (2002) noted that insufficent aration of rtifcl growing mdia is a comon proble in greenhous production. The soiles mdi amendmnt in question is a product of Azadirachta indica A. Juss, or t nem tre. The ne tree is known as the ?vilage dispensary? in Indi and Southeast Aia, where its native (Biswa et al., 2002). This i due to the result of both scienc and tradition about the medicinal and agricultural uses that neem provides. The nem tre is an evergreen tre belonging to the mahogany faily, Mliac, of which Melia azedarach Chinaberry is alo a member (Bisw et al., 2002). Steped in Indian tradition and lore, the neem tree continues to play roles in their traditions, mdicine and agriculture and now t rest of the world is beginning to pay more atention to neem, whih has been somewhat controversial to the traditionalist in Asia (Mrden, 1999). Nm has ben usd as a soil amendmnt in many studies in Indi. Nem sed residue provided a nitrogen value, aftr oil extraction, of 7% and at a relas rat fast enough to satisfy maize nutrition (Agbenin et al., 1999). Nem oil aso enhanced plnt growth when incorporatd into soil at 2.5, 5.0 and 20 ppm, though 10 ppm decreasd plant growth (Bhaskar and Charyulu, 2005). Likewise, Agyarko et al. (2006) reported tha soil nutritional levels increased with poultry manure and increasing levels of nem leaves. Bhala and Dvi Prasd (2008) reported higher vegetive growth in plants than reproductive growth (both showing higher growth than the control) when neem cke was incorporated into the soil. Nm?s ipact wihin a substrae has been atributed to its potential urese retardation activiy (B. Hurst, personal communication). Ureas i the naturaly occurring enzyme in soil responsible for the hydrolysis of ure into carbon dioxide and 17 amonia (Mnunza et al., 1997). Amonium and amonia are the nitrogen sources within fertilzrs (be it synthetic fertilzr or organic), but the amonia cn undergo volatiltion, which may dereas nitrogen available to t plant. Therefore, it is often desirble to have certin levels of urese inhibition in order to constrain amonium production, resulting in a more readily available supply of nitrogen over a longer period of time. The aasing of amonium cn ld to problems other than amonia volatilztion, such as nitrie toxicity and damage to young plants (Brener and Douglas, 1971). Mohanty et al., (2007) reported on the potential inhibitory efects of neem sd kernel powder on urease in thre mineral soils native to India, showing slight suppresion of urease activiy when applid to acidi soils. M?ndez-Bautist e al., (2009) studied the efcts of nem leaf extracts on greenhouse gas emisons and inorganic nitrogen in urea- amnded soil and reported that the laf extract had no significant efet on urease, but y lit nirificaton. Majumdar et al., (2000) coated ure with nem sd powder before adding to rie filds in North India, resulting in slight nitrificaton inhibition. Kumar et al., (2007) usd neem oils to coat urea and added it to sndy-loam soils resulting in some nitrificaton inhibition as wel. Bhala and Devi Prasad (2008) showed, in one of their studis, that addition of nem cake into a mineral soil is an economical and efctive method for reducing fertilzr applicton by prolonging fertilzr availble to the plant. The objective of this experiment was to determine the impact of neem cake powder as substra component on plant growth or quality. 18 Materials and Methods: Two stock substraes wre used for this study: one with poultry protein compost (PPC) as the fertilzr sourc and the otr with a strter charge fertilzr (a cust 7-2 10 blend) incorporated into t mixture. The compost sock mixture consisted of pinebark, poultry protin compost, perlite and vermiculte in a 50:17:10:5 ratio. To acount for the volume of the PPC in the first mixture, peat ws used as a substiute. With t two stock substraes on hand, neem cke powder ( residual of the nem tree sds or kernels after being cold-presed for oil) was incorporated at 1, 2, 3, 4 and 5 percnt by volum whil aso keeping a ?control? tretment with no nem added. Due to neem cake?s sal particle siz and the relatively negligibl aounts that were usd, it ws not necesry to compensate for its addition by the subtraction of any of the otr amendmnts. The neem ws thoroughly incorporated into the mixtures a t starter charge was added to t non-compost sock substra. Thereere twelve treatments (six percentages of neem and two stock substraes) replicated 10 tims for a totl of 120 experimntl units. Square pots (4?x 4?x 6?) wre fild with each substrae and planted with blue salvi (Salvia farinacea ?Fr Rhea Blue?). Pots were arranged in a randomizd complet block design and wtred on an ?s neded? basi. Clear wter was used on al treatments for the first wek and on the compost treatmnts for the duration of the study. Non-compost treatmnts, however, reeived fertigaton three tims a wek and clear wter every fourth wtering aftr the inital wk of instaltion with 20-20 solubl frtilzr (TotalGro ? , Inc, Winnsboro, Louisina) at 175 ppm nitrogen. Percnt nitrogen sources for this fertilzr are 5.98% nitrae nitrogen, 5.58% amoniacl nitn and 8.44% urea nitrogen. The study was 19 conducted in a double-poly greenhouse for the 8-wek study with he trmosta set a min/ax tmperatures of 65/78 o F. Dt taken 28 and 56 days after planting (DAP) included: growth indices, pH, elctrial conductiviy (EC), SPAD-502 (mesure chlorophyll levels in the plants) redings nd shoot dry weight (shoot dry wights was measured at 56 DAP; t shoot fresh cutti were placd in a 175 o F oven for four days and weighe). Al data were analyzed using Wlr-Duncan k-ratio t-tes in SAS software. Results: Mid-study (28 DAP): There were fw diferencs among al treatments 4weks into the st (Table 1). Non-compost treatmnts without nee and with 1% ne had significantly higher growth indies than any of the PC treamnts. The 4% neem, non- compost tretment also had significantly higher growth indices than did t 0-4% ne t treatnts, but GI?s were not significantly greatr n the 5% neem, PC treament. Masurements of pH at 28 DAP show that the one percent nem, non-compost mixture had the lowst and was not significantly diferent from the 2, 3 and 5% neem, non-compost mixtures (Table 1). It ws, however, significantly lower than al other mixtures. Electrial conductiviy shows that the zro percnt, PPC mixture was significantly higher in EC than 4 and 5% neem, PPC ixtures a wel as the 0%, non- compost mixture (Table 1). Al other mixtures are not significantly difrent. 20 SPAD result show that the 0% neem and 3% neem, non-compost mixtures had significantly higher SPAD readings than did 0, 2 and 4%, cpostixtures (Table 1). The rest of the treatments had no significant diferenc in values. erination (58 DAP): At terminaton, result show that one percent nem in the non-compost mixture had significantly greater GI?s than any of the PPC ixtures, a wel as the 4 and 5% neem, non-ompost tretmnts (Table 2). T 2% neem, non-compost mixture also had significantly greater GI?s than the 0, 1, 2, 3 and 5% ne, PPC ixes a wel as the 5% neem, non-ompost tretment. Within t PPC treaments, there were no significant difrencs in GI. SPAD result show that the 2 to 5% neem, non-compost mixtures had significantly higher SPAD readings than did any of the PPC treaments and the 1% neem, non-ompost mix (Table 2). In addition, the 5% neem, non-compost treatent was alo significantly gretr than the 0% neem, non-compost treatent at 56 DAP. Mesurements for pH show that the 4% neem, non-compost mixture had a pH significantly lowr than any of the PPC treants a wl as the 0% neem, non-compost mix (Tble 2). The 0and 2% neem, ixes, likewise, had a significantly higher pH than al non-compost mixtures excpt the 0% neem, non-compost tretment. At 56 DP, the 4and 5% neem, non-compost mixtures had signifiantly greater EC masureents than t 0-2% , PPC ixtures and the 0% neem, non-compost tretment (Tbl 2). In the same way, the 0% neem, non-compost treatnt had a significantly lowr EC value than the 1, 3, 4 and 5% ne, non-t tretments. Shoot dry weights of0 and 1% neem, PPC treants were significantly lower than any of the non-compost tretnts (Tabl 2). The 2% nem, PPC trement was 21 also significantly less than al non-compost treatments except for the 4% neem, non- compost mix. In the same way, the 0-3% ne, non-compost treatents had significantly greter dry wights than al PPC mixes, except for the 5% nem, PPC trement. Discuion: N-compost treatments, those containing peat and fertigated, outperformed the PPC treaments. A a whol, the non-ompost tretments had greatr growth indics, higher SPAD readings and greater fresh and dry wights. Within the non-compost treaments, the retments with ls nem (especialy 0 and 1%) often outperformed the other tretnts within the non-compost group. Within the PPC group, though, t opposite was true: t 5% neem with PPC treament outperformed al other mixes within the PC group. It is dificult to draw one overal conclusion from this experiment, especialy since the initial purpose of this study was to consider the efect of ne at diferent percntges. It sms, howver, that the PPC used in 6 of t treatments may hinder the rformance of the plants when compared to the superior result of plnts in the fertigatd, non-compost tretments. Examining t neem percentages sems to render a twofold conclusion: when usd in conjunction with PPC, higher ne percntages (in this cae 5% neem) aid t plant in growth; but, within t fertigated, non-compost tremnts, litl-tono nee outperformed the higher percntages of nem (0 and 1% nee, in this cae, outperformd 4 and 5% nem treaents within the non-compost group). Consquently, when using poultry protein compost, t addition of neem has a 22 benefical efect on the plant, however it has an antagonistic efet wih increasing amounts added as an amendmnt in the case of frtigated treatmnts. 23 Literatue Cited: Agbenin, J.O., E.B. Agbaji, I. Suleiman, and A.S. Agbaji. 1999. Asemnt of nitrogen mineralizaton potential nd availbiity from neem sd residue in a savanna soil. Biol. Fertil. Soils 29:408-412. Agyarko, K., P.K. Kwakye, M. Bonsu, and B.A. Osei. 2006. Impact of applicaton of neem laves and poultry manure on nutrint dynamics of a Hapli Arisol. Archis of Agronomy and Soil Scienc 52:687-695. vent, T. 2003. So You Want to Start a Nursry. Timber Pres, Inc. Bhala, R.S. and K.V. Devi Prasd. 2008. Ne cake-urea mixed applicatons increase growth in paddy. Current Scienc 94:1066-1070. Bhaskar, K.V. and P.B.B.N. Charyulu. 2005. Efect of environmental fctors on nitrifying bacteria isolted from the rhizosphere of Setaria itali (L.) Beauv. African J. of Biotchnology 4:1145-1146. Bilderbak, T.E., W.C. Fonteno, and D.R. Johnson. 1982. Physical properties of media composed of peanut hulls, pine bark, and peatmoss and their efcts on azl growth. J. Amer. Soc. Hort. Sci. 107:522-525. Biswa, K., I. Chatopadhyay, R.K. Banerje, and U. Bandyopadhyay. 2002. Biological ctivies and medicinal propertis of nem (Azadirachta indica). Current Sinc 82:1336-1345. Boyer, C.R., G.B. Fain, C.H. Gilam, T.V. Galgher, H.A. Torbert, and J.L. Sibley. 2008. Clen Chip Residual: A substrae component for growing annuals. HortTchnology 18:423-432. 24 Bremner, J.M. and L.A. Douglas. 1971. Inhibition of urease activiy in soils. Soil Biol. Biochem. 3:297-307. Chang, C.P. and S.M. Lin. 2007. The formation and growing properties of poly (ethylene trephthalate) fiber growing media after hermo-oxidative treatment. Matrils Scienc nd Engineering 457:127-131. Cole, D.M., J.L. Sibly, E.K. Blythe, D.J. Eakes, and K.M. Tilt. 2002. Evaluation of cotton gin compost as horticultural substrae. SNA Research Conferenc 47:274-277. Evans, M.R. and M.M. Gachukia. 2007. Physical properties of sphagnum peat-based root substraes amnded with perlite or parboild fresh rice hulls. HortTechnology 17:312-315. Fain, G.B., C.H. Gilam, J.L. Sibley, and C.R. Boyer. 2008. Establishment of greenhouse-grown Tagets patula and Petunia xhybrida in ?WholTr? substras. Acta Hort. 782:387-393. Handreck, K.A. 1983. Prticle siz and the physical properties of growing media for ontainers. Commun. In Soil Sci. Plant Anal. 14:209-222. Jackson, B.E. and R.D. Wright. 2007. Pine tree substrae: Frtilty requirements. SNA Research Conferenc 52:523-526. Lu, W., J.L. Sibly, C.H. Gilam, J.S. Bannon, and Y. Zhang. 2006. Estimation of U.S. bark generation and iplictons for horticultural industries. J. Environ. Hort. 24:29-34. 25 Kumar, R., C. Devakumar, V. Sharma, G. Kakkar, D. Kumar, and P. Panneerslvam. 2007. Influenc of physiochemicl paraetrs of nem (Azadirachta indica A. Juss) oil on nitrificaton inhibition in soil. J. Agric. Food Che. 55:1389-1393. Majumdar, D., S. Kumr, H. Pthak, M.C. Jain, nd U. Kumar. 2000. Reducing nitrous oxide emison from an irrigated ric field of north Indi with nitrificaton inhibitors. Agriculture, Ecosystms and Environment 81:163-169. Manunz, B., S. Deiana, M. Pintore, V. Solinas, and C. Gsa. 1997. A molecular modeling study of the urease active site. J. of Moleculr Structure (Theocm) 419:33-36. Marble, S.C., C.H. Gilam, J.L. Sibley, G.B. Fain, H.A. Torbert, T.V. Galgher, and J.W. Olive. 2010. Evaluation of composted poultry liter as a substrae amendmnt for WholeTre, Clean Chip Residual, and pinebark for container grown woody nursry crops. J. Environ. Hort. 28:107-116. Marden, E. 1999. The Nem Tree Patnt: International Conflict over the Commodificaton of Life. Boston Collge International and Comparative Law Review 22:279-295. M?ndez-Bautista, J., F. Fern?ndez-Luque?o, F. L?pez-Valdez, R. Mendoza-Cristino, J.A.ontes-Molina, F.A. Gutierrez-Miceli, and L. Dendooven. 2009. Efect of pest crolling nem (Azadirachta Indica A. Juss) and mat-raton (Glirdia sepium Jacquin) laf extracts on emison of greenhouse gas and inorganic-N contnt in ure-mended soil. Cheosphere 76:293-299. 26 Mitchel, C.C. and J.O. Donald. 1995. The value and use of poultry manures a fertilzr. Alabam Cooperative Extension System. Noveber 1995. Retrieved from: http:/w.aces.edu/pubs/docs/ANR-0244/ Mohanty, S., A.K. Patra, and P.K. Chhonkar. 2007. Nem (Azadirachta indica) seed kernel powder retrds urease and nitrificaton activies in diferent soils at contrasting moisture and tmperature regims. Bioresourc Tchnology 99:894- 899. Nemati, M.R., J. Caron, O. Banton, and P. Tardif. 2002. Detrmining air entry value in pet substraes. Soil Sci. Soc. Am. J. 66:367-373. Nkongolo, N.V. and J. Caron. 2006. Pore space organization and plant response in peat substraes: I. Prunus xcistena and Spiraea japonic. Sientfic Resarch and Essay 1:77-89. Noguera, P., M. Abad, R. Puchades, A. Maquieira, and V. Noguera. 2003. Influence of particle siz on physical and chemicl properties of coconut coir dust as container mdium. Comm. in Soil Si. and Plant Analysis 34:593-605. Paul, J.L. and C.I. Le. 1976. Relation betwen growth of chrysanthemums and aeration of various container medi. J. Amr. Soc. Hort. Sci. 101:500-503. Schw?rzel, K., M. Renger, R. Sauerbrey, and G. Wesolk. 2002. Soil physical haractristic of pet soils. J. Plant Nutr. Soil Sci. 165:479-486. Smith, C.A. and D.A. Hal. 1994. The development of perlite as potting substrae for ornaental plnts. Act Horticultura 361:159-166. Smith, C.A. P. McDonald, and A.M. Sword. 1995. Culture of hardy woody ornamentals in pure-mineral ontiner substraes. Acta Horticulturae 401:161-167. 27 Urrestarazu, M., G.A. Mart?nez, and M.C. Sals. 2005. Almond shel waste: possible locl rockwool substiute in soiles crop culture. Scienta Horticultura 103:453- 460. Wels, D.E. 2008. Evaluation of spent tea grinds asn alternative horticultural substrae component. Thesi. Auburn University. 28 29 30 Chapter III Gas Production from Soiles Mdia Amende with Nem Cake Powder Signficane to Industry: Nursry and greenhouse growers continue to sek materils to decrease costs of plant production whil maintning nvironmental stwrdship. Incorporation of neem cke as substrae component could potntily impact nirogen releas a result of altering stra bactrial activiy. This prelinary study investigas the impact of nem on substrae gas reles and provides a starting point to further investigaon regarding ne us as substrae component. In this study, three substrae groups were tesd with varying percentages of nem, and this paper reports on both across-group result as wel as within-group result. Across al three substraes, three percnt neem within the pine bark + poultry protein compost + nem group was significantly greatr in CO 2 production than al treatmnts within the pine bark + nee group as wel as zero percent nem within its own group and t pi bark + poultry protein compost + 19-612 + ne group. Nitrous oxide emison was significantly greatr in the pine bark + poultry protein compost + 19-612 + nee group than al other tretments. Within-group comparisons reveal that hree percnt nem had greater CO 2 eison than zero percent neem for both the pine bark + ne and pine bark + poultry protein compost + nem groups. Three percnt neem also produced significantly greatr CH 4 than zero percent neem in the pine bark + poultry protin compost + neem group ? and within the sam 31 substrae group, two percent nem had significantly greater N 2 O emison than zero percnt nem. Comparing within-group, there were no significant differencs among treaents in the pine bark + poultry protein compost + 19-612 + ne group for al thre gas analyzed. Introduction: Fertilzr is an expensive part of any plant production program and environmental safty is becoming an increasingly importnt subject. Therefore, any cost-efctive method to reduce frtilzer us in an environmntaly-friendly manner is a neded and valuabl product. Nitrogen is often viwed as limting factor in plnt nutrition, and while there a many forms or sourcs of nitrogen, our study focused specifaly on urea. Urea breks down into amonium with the aid of an enzyme known as urese. Amonium then further breaks down into amonia, which then undergoes volatilztion. Therefore, slowing down this ctlysis of ure could, in tory, prolong substrae nitrogen supplies. Since urease in soil is a byproduct of bacteria, limting ureas production by afcting the enzym itself or its bacterial producrs could inhibit the breakdown of urea. Beginning in the 1970?s, pine bark quickly became an invaluable resource to the horticulture industry. Present-day demands for this comodity, howver, have also increased and, with the growth of the horticulture sector, our industry is feling the presure to find some altrnatives to exclusive pine bark usage (Avent, 2003). While greenhouse plants arelmost exclusily produced in pet-substraes, 75-100%, by volum, of continer substraes in the eastern US are comprisd of pine bark (Lu et al., 32 2006). Future projections project the rising cost of pine bark combined with les availbiity to the hortiulture industry (Lu et al., 2006). Numerous organic and inorganic substances are being used to develop varying substras. Clan chip residual (the materil lft on the forest floor following timber harvest) cn be a relistc alternative to pine bark-exclusive substraes for ornaental plant production (Boyer et al., 2008). Fain et al. (2008) succesfully usd WholTre (whole pine tree shoots) in varying percentges a suitabl grenhouse mdia for growing marigolds and petunias. With additional frtilzer, Jckson and Wright (2007) were able to grow plants in 100% pulverized pine wood. Cotton gin compost was used as a viabl substrae component by Col t al. (2002). Spent tea grinds can be succsfully used in horticultural substraes (Wls, 2008). Poultry litr ws a suitble amndment for substraes omprisd primarily of either pine bark, WholeTre (creatd by chipping, then further milng entire pine trees ? log, limbs, needls and bark), or clen chip residual for growing plants in contairs with favorabl result and is a readily available source of nitrogen (Fin et al., 2008; Mrble et al., 2010; Mitchel and Donald, 1995). Parboiled fresh rice hulls can be used sucesfully as a substrae component comparabl to perlit (Evans and Gchukia, 2007). Using stras from peanut hulls, pine bark, peatmoss and combination thereof, Bilderback et al. (1982) reported succes in growing azles. In searching for an ecologicaly-friendly alternative to rockwool, Urrestarau et al. (2005) concluded that almond shels can be an efcti and benefical soiles mdia for plant production. Coconut coir c a vible substrae for plnt production, however its succes depends on particle siz (Noguera et al., 2003). Peat- vermicult substras srved as viable substras for growing chrysanthemums (Pul and 33 Le, 1976). Mineral substraes have the potential to perform simlarly to peat-based mixes (Sith e al., 1995). Smith and Hal (1994) determined that a perlite-sd substrae can be comparable in manageent and productiviy to peat-basd potting mixes. While potntial aternative substraes sm to abound, there a requirements that must be mt of the mtril in order for it to be usable and efective. Chang and Lin (2007) report that basic requirements for a succsful and benefial plnt-growing medium include: excelnt cheical resitance properties, light weight, inexpensivenes, abs of pest and disae and availbiity Nkongolo and Caron (2006) noted that particle siz, specifly in pet and pine bark-based substraes, influencs plant respons. Handrek (1983) reports the importanc of particl siz and advise that substrae formultors heed the ?fines? fraction of a substrae, especialy particl size smalr than 0.5mm, as thi size controls the physical propertis of pine bark-basd substraes. Shrinkage is a physical property of organi soils (such as peat) that must be managed (Schw?rzel et al., 2002). Nemati e al. (2002) noted that insufficent aration of rtifcl growing mdia is a comon proble in greenhous production. The soiles mdi amendmnt in question is a product of Azadirachta indica A. Juss, or t nem tre. The ne tree is known as the ?vilage dispensary? in Indi and Southeast Aia, where its native (Biswa et al., 2002). This i due to the result of both scienc and tradition about the medicinal and agricultural uses that neem provides. The nem tre is an evergreen tre belonging to the mahogany faily, Mliac, of which Melia azedarach Chinaberry is alo a member (Bisw et al., 2002). Steped in Indian tradition and lore, the neem tree continues to play roles in their traditions, mdicine and 34 agriculture and now the rest of the world is beginning to pay more atention to neem, whih has been somewhat controversial to the traditionalist in Asia (Mrden, 1999). Nm has ben usd as a soil amendmnt in many studies in Indi. Nem sed residue provided a nitrogen value, aftr oil extraction, of 7% and at a relas rat fast enough to satisfy maize nutrition (Agbenin et al., 1999). Nem oil aso enhanced plnt growth when incorporatd into soil at 2.5, 5.0 and 20 ppm, though 10 ppm decreasd plant growth (Bhaskar and Charyulu, 2005). Likewise, Agyarko et al. (2006) reported tha soil nutritional levls increased with poultry manure and increasing levels of nem leaves. Bhala and Dvi Prasd (2008) reported higher vegetive growth in plants than reproductive growth (both showing higher growth than the control) when neem cke was incorporated into the soil. Nm?s ipact wihin a substrae has been atributed to its potential urese retardation activiy (B. Hurst, personal communication). Ureas i the naturaly occurring enzyme in soil responsible for the hydrolysis of ure into carbon dioxide and amonia (Mnunza et al., 1997). Amonium and amonia are the nitrogen sources within fertilzrs (be it synthetic fertilzr or organic), but the amonia cn undergo volatiltion, which may dereas nitrogen available to t plant. Therefore, it is often desirable to have certin levels of urese inhibition in order to constrain amonium production, resulting in a more readily available supply of nitrogen over a longer period of time. The aasing of amonium cn ld to problems other than amonia volatilztion, such as nitrie toxicity and damage to young plants (Brener and Douglas, 1971). 35 Mohanty et al., (2007) reported on the potential inhibitory efects of neem sd kernel powder on urease in thre mineral soils native to India, showing slight suppresion of urease activiy when applid to acidi soils. M?ndez-Bautist e al., (2009) studied the efcts of nem leaf extracts on greenhouse gas emisons and inorganic nitrogen in urea- amnded soil and reported that the laf extract had no significant efet on urease, but y lit nirificaton. Majumdar et al., (2000) coated ure with nem sd powder before adding to rie filds in North India, resulting in slight nitrificaton inhibition. Kumar et al., (2007) usd neem oils to coat urea anddded it to sndy-loam soils resulting in some nitrificaton inhibition as wel. Bhala and Devi Prasad (2008) showed, in one of their studis, that addition of nem cake into a mineral soil is an economical and efctive method for reducing fertilzr applicton by prolonging fertilzr availble to the plant. The objectives of this study were to determine neem cake powder?s efect on gas production and emison from soils mdia. This wa done in hope to extrapolat the result into the potential prolongation of nitrogen sourcesithin the substraes. Materials and Methods: The study consisted of three groups of treatments: 100% pine bark (PB) + neem; 80% PB + 20% poultry protin compost (PPC) + ne; and 80% PB + 20% PC + urea + neem. Within each of these ubstraes wre varying concentrations of neem. Within the PB + nee substra wre four treamnts with 0, 1, 2 and 3% nee. The PB + PC + neem substra had the same percntges of nee added, but also included 20 percent PPC with 80 percent PB. T third substra contained the same percntages of PB 36 (80%) and PPC (20%) as the second substrae, and with the varying neem concentrations, but with the addition of Sott?s Osmocot Clasic 19-612 at 9 lbs/yd 3 (1.7 lbs N/yd 3 ). Each of the 12treaents contained 4 repliates for a total of 48 experimental units. Substraes wre plcd in trade-galon continers without plnts and plcd in a glas greenhous at the USDA Soil Dynamics Laboratory, Auburn University, Alabam. Substras wreatered as needed, but with minil eaching. Moist conditions were necesary to mimc rhizosphere microenvironments in order to facilte microbial growth. Dt were taken at regular intervals beginning in My 2010 and ended in August 2010. Dats collcted 3 days weekly for the first 2weks and then once weekly for the next 7 weks. Aftr that, data ws collectd bikly. In order to detrmine substra microbial activiy, gas emisons wre ollectd from an airtght gas chamber large enough to aommodate one pot each. The top of the gas chamber was outfited with a rubber septum through which a needl could penetrae. Four evacuated collction vials were neded for each experimntal unit, each one represnting a tim within the 15 minuts of collection (ties 0, 1, 2 and 3 represent initial tme and 5, 10 and 15 minutes, respectively). Gas smpls wre pulled for each experintl unit for each of the aforementioned ties and resultre analyzed using a gas chromtograph. Gas spls ttd for wre: carbon dioxide (CO 2 ), mthane (CH 4 ) and nitrous oxide (N 2 O), which represent microbil respiration.. CO 2 , CH 4 and N 2 O data were analyzed using Tukey?s Studentizd Range Test in SAS Staistcl Softwre (alpha = 0.05). 37 Results: Overall: Notation for reporting data wil adhere to the following guidelines: PB is pine bark; PPC is poultry protein compost; fertilzr wil refer to the Osmocote 19-6 12; and when entire groups of treatmnts are refrenced, the values that follow are given in order of neem rate increase within the group?s treatmnts. T unit for trace gas emison values i ?ols m -2 in -1 . Al data is preentd in Table 3. First, result wil be given for across-ubstra comparisons. Three substraes tesd in this experimentre: 100% PB + neem; 80% PB + 20% PPC + nem and 80% PB + 20% PPC + 19-612 + neem. Secondly, result wil be given for within-substrae treaments. Across-ubstrate rsult: Carbon Dioxde (CO 2 ): Increasing neem percentage (by volume) as a ubstrae component appeared to increase CO 2 production (Tbl 3). Howver, in the PB + nem treaments, there was no stistcl diferenc among treatents. Within t PB + PPC + ne group, CO 2 production for the 3% ne treatent was staitcly larger than the 0% neem treatent. However, tre was no stistcl difrence among tretents in t PB + PC + frtilzr + nem group. Across al groups, PB + PPC +3% neem had the highest value for CO 2 production, though not staistcly difrent tn PB + PPC + 1 and 2% neem. The PB + 0% neem treatent had the lowest value for CO 2 , but was not stistcly difrent than any of the PB + nem treatnts, PB + PPC + 0% neem, or PB + PPC + fertilzr + 0% neem. 38 Methane (CH 4 ): Methane?s relation to neem percentage does not sem to be as clear-cut as with CO 2 . Thre percent ne usd in conjunction with PB + PPC had signifiantly higher CH 4 than 0% nem in the same mixture (Table 3). There was no sificnt diferenc in 4 levels aong tretnts within the otr two groups testd. Again, among al groups PB + PPC + 3% neem had t highest CH 4 value, but was not significantly diferent than PB + 0 and 2% ne; PB + PPC + 1 and 2% neem or PB + PPC + fertilzr + 1, 2 and 3% neem. The PB + PPC + 0% neem had the lowst value for CH 4 across al treatments, but was not staistcly difrent from any treatment other than PB + PPC + 3% ne. Nitrous Oxide (N 2 O): For nitrous oxide there were no staistcl diferencs among treatents within the PB + neem group or t PB + PPC + neem group (Table 3). The PB + PPC + frtilzer + nee group, though, shows that N 2 O eison for 3% nem was significantly higher than 0 and 1% neem. Aross al treatments, N 2 O from 3% ne in PB + PPC + fertilzr was significantly greter han al other treatments, other than 2% nem in PB + PPC + fertilzr. N 2 O from PB + PPC + fertilzr + 2% neem was greter than al treatments from the PB + neem and P + nem groups. N 2 O from the 0 and 1% ne treaments within the PB + PB + fertilzr group were also staistcly greater han al trentsithin t PB + nem and PB + PB + nem groups. Figure 3 also shows a stepwis increase in N 2 O production with the increase of nee within each substrae, though there a negligible result in t pine bark only mixes. 39 Within-substrate rsult: PB + nem: Three percnt neem within the pine bark media had significantly greater CO 2 eison than zro percnt ne (Table 4), though neither t 3nor 0% nem tretnts were significantly diferent than the 1 and 2% neem treatents. There were no significant difrences among treatnts in this substra group for CH 4 and N 2 O mison. PB + PPC + neem: The 3% neem treaent within the PB + PPC + neem substrae group had significantly greatr CO 2 eison than t 0% ne treatnt of the sam group (Table 5). Neither t 3 nor 0% neem treatents wre significntly diferent than the 1 and 2% nem treatnts. Methane result wre simlar to CO 2 : 3% nem was significantly greater than 0% neem, though neither t 3 nor 0% nee treatents were sifintly difrent tn the 1 and 2% neem treatents. Two percnt nem had significantly greater N 2 O production than 0% ne, though neither had significantly diferent values from 1 and 3% neem. PB + PPC + 19-612 + ne: Aong al treatments and for al three gas reults, CO 2 , CH 4 and N 2 O, there were no significant diferencs (Tabl 6). Discuion: Aross ubstraes, there were fw diferencs among treatents. Three percnt neem in both the PB + PPC + nem and PB + PPC + 19-612 + nem groups had greater CO 2 production than 0-2% nee in the PB + neem group. Three percnt nee in the PB + PPC + nem produced more CO 2 than al P treatmnts a wl as 0% neem within its own group and 0% neem in the PB + PPC + 19-612 + nee. Methane 40 comparisons show no notable result across ubstraes. The PB + PPC + 19-612 + neem group, as a whole, had significntly greater N 2 O production than any other treatnts. Within-group comparisons show that 3% neem within the PB + neem and PB + PPC + neem groups had significantly greater CO 2 production than 0% ne within their groups. Thre percent nem within the PB + PPC + neem group also had significantly greater CH 4 production than 0% nee within the sa group. PB + PPC + 2% neem ws significntly greater in N 2 O emison than 0% neem, as wel. Across al treatnts and for al three gas tstd (CO 2 , CH 4 and N 2 O), the PB + PPC + 19-612 + neem group had no significant diferencs. Studies to detrmine the fate of urease when neem is added are ongoing, with some supplmntal dat not hving ben analyzd yet. It ss reaonable to conclude tha basd on the presentd data, neem does have an efect on soil respiration, though more testing to prove the extent to which this occurs is currently underway. Current tsing includes t aformntioned aid-oated tubes for amonia voltilztion, pH and EC, as wel as nutrient compositon of the difrent tatents. 41 Literatue Cited: Agbenin, J.O., E.B. Agbaji, I. Suleiman, and A.S. Agbaji. 1999. Asemnt of nitrogen mineralizaton potential and availbiity from neem sd residue in a savanna soil. Biol. Fertil. Soils 29:408-412. Agyarko, K., P.K. Kwakye, M. Bonsu, and B.A. Osei. 2006. Impact of applicaton of neem laves and poultry manure on nutrint dynamics of a Hapli Arisol. Archis of Agronomy and Soil Scienc 52:687-695. vent, T. 2003. So You Want to Start a Nursry. Timber Pres, Inc. Bhala, R.S. and K.V. Devi Prasd. 2008. Ne cake-urea mixed applicatons increase growth in paddy. Current Scienc 94:1066-1070. Bhaskar, K.V. and P.B.B.N. Charyulu. 2005. Efect of environmental fctors on nitrifying bacteria isolted from the rhizosphere of Setaria itali (L.) Beauv. African J. of Biotchnology 4:1145-1146. Bilderbak, T.E., W.C. Fonteno, and D.R. Johnson. 1982. Physical properties of media composed of peanut hulls, pine bark, and peatmoss and their efcts on azl growth. J. Amer. Soc. Hort. Sci. 107:522-525. Biswa, K., I. Chatopadhyay, R.K. Banerje, and U. Bandyopadhyay. 2002. Biological ctivies and medicinal propertis of nem (Azadirachta indica). Current Sinc 82:1336-1345. Boyer, C.R., G.B. Fain, C.H. Gilam, T.V. Galgher, H.A. Torbert, and J.L. Sibley. 2008. Clen Chip Residual: A substrae component for growing annuals. HortTchnology 18:423-432. 42 Bremner, J.M. and L.A. Douglas. 1971. Inhibition of urease activiy in soils. Soil Biol. Biochem. 3:297-307. Chang, C.P. and S.M. Lin. 2007. The formation and growing properties of poly (ethylene trephthalate) fiber growing media after thermo-oxidative treatment. Matrils Scienc nd Engineering 457:127-131. Cole, D.M., J.L. Sibly, E.K. Blythe, D.J. Eakes, and K.M. Tilt. 2002. Evaluation of cotton gin compost as horticultural substrae. SNA Research Conferenc 47:274-277. Evans, M.R. and M.M. Gachukia. 2007. Physical properties of sphagnum peat-based root substraes amnded with perlite or parboild fresh rice hulls. HortTechnology 17:312-315. Fain, G.B., C.H. Gilam, J.L. Sibley, and C.R. Boyer. 2008. Establishment of greenhouse-grown Tagets patula and Petunia xhybrida in ?WholTr? substras. Acta Hort. 782:387-393. Handreck, K.A. 1983. Prticle siz and the physical properties of growing media for ontainers. Commun. In Soil Sci. Plant Anal. 14:209-222. Jackson, B.E. and R.D. Wright. 2007. Pine tree substrae: Frtilty requirements. SNA Research Conferenc 52:523-526. Lu, W., J.L. Sibly, C.H. Gilam, J.S. Bannon, and Y. Zhang. 2006. Estimation of U.S. bark generation and iplictons for horticultural industries. J. Environ. Hort. 24:29-34. 43 Kumar, R., C. Devakumar, V. Sharma, G. Kakkar, D. Kumar, and P. Panneerslvam. 2007. Influenc of physiochemicl paraetrs of nem (Azadirachta indica A. Juss) oil on nitrificaton inhibition in soil. J. Agric. Food Che. 55:1389-1393. Majumdar, D., S. Kumr, H. Pthak, M.C. Jain, nd U. Kumar. 2000. Reducing nitrous oxide emison from an irrigated ric field of north Indi with nitrificaton inhibitors. Agriculture, Ecosystms and Environment 81:163-169. Manunz, B., S. Deiana, M. Pintore, V. Solinas, and C. Gsa. 1997. A molecular modeling study of the urease active site. J. of Moleculr Structure (Theocm) 419:33-36. Marble, S.C., C.H. Gilam, J.L. Sibley, G.B. Fain, H.A. Torbert, T.V. Galgher, and J.W. Olive. 2010. Evaluation of composted poultry liter as a substrae amendmnt for WholeTre, Clean Chip Residual, and pinebark for container grown woody nursry crops. J. Environ. Hort. 28:107-116. Marden, E. 1999. The Nem Tree Patnt: International Conflict over the Comodificaton of Life. Boston Collge International and Comparative Law Review 22:279-295. M?ndez-Bautista, J., F. Fern?ndez-Luque?o, F. L?pez-Valdez, R. Mendoza-Cristino, J.A.ontes-Molina, F.A. Gutierrez-Miceli, and L. Dendooven. 2009. Efect of pest crolling nem (Azadirachta Indica A. Juss) and mat-raton (Glirdia sepium Jacquin) laf extracts on emison of greenhouse gas and inorganic-N contnt in ure-mended soil. Cheosphere 76:293-299. 44 Mitchel, C.C. and J.O. Donald. 1995. The value and use of poultry manures a fertilzr. Alabam Cooperative Extension System. Noveber 1995. Retrieved from: http:/w.aces.edu/pubs/docs/AR-0244/ Mohanty, S., A.K. Patra, and P.K. Chhonkar. 2007. Nem (Azadirachta indica) seed kernel powder retrds urease and nitrificaton activies in diferent soils at contrasting moisture and tmperature regims. Bioresourc Tchnology 99:894- 899. Nemati, M.R., J. Caron, O. Banton, and P. Tardif. 2002. Detrmining air entry value in pet substraes. Soil Sci. Soc. Am. J. 66:367-373. Nkongolo, N.V. and J. Caron. 2006. Pore space organization and plant response in peat substraes: I. Prunus xcistena and Spiraea japonic. Sientfic Resarch and Esay 1:77-89. Noguera, P., M. Abad, R. Puchades, A. Maquieira, and V. Noguera. 2003. Influence of particle siz on physical nd chemicl properties of coconut coir dust as container mdium. Comm. in Soil Si. and Plant Analysis 34:593-605. Paul, J.L. and C.I. Le. 1976. Relation betwen growth of chrysanthemums and aeration of various container medi. J. Amr. Soc. Hort. Sci. 101:500-503. Schw?rzel, K., M. Renger, R. Sauerbrey, and G. Wesolk. 2002. Soil physical haractristic of pet soils. J. Plant Nutr. Soil Sci. 165:479-486. Smith, C.A. and D.A. Hal. 1994. The development of perlite as potting substrae for ornaental plnts. Act Horticultura 361:159-166. Smith, C.A. P. McDonald, and A.M. Sword. 1995. Culture of hardy woody ornamentals in pure-mineral ontiner substraes. Acta Horticulturae 401:161-167. 45 Urrestarazu, M., G.A. Mart?nez, and M.C. Sals. 2005. Almond shel waste: possible locl rockwool substiute in soiles crop culture. Scienta Horticultura 103:453- 460. Wels, D.E. 2008. Evaluation of spent tea grinds asn alternative horticultural substrae component. Thesi. Auburn University. 46 47 48 49 50 Chapter IV Final Discuion To facilte the final discusion chapter, the discusion wil be segmnted acording to the experiments deribed within ts tis. Commercially Availabe Organic Substrate Comparison: It was during this experiment that we originaly noticed the presenc of diferent fungi in some of the Jungle Growth ? products. The fungi?s presnc did not presnt a concern, it was almost t opposite ? w asumed that increased microorganismal activiy within the potting media could very wl benefit plant mtril. It did concern us, though, that presnc of the fungi was sporadic. Within any given Jungl Growth ? product, there was bag-tobag varition whih also showed in the performance of the t. Also, it was noted that some of the Jungle Growth ? products continued to undergo a ?het? aftr stocking. This could have ben due to the poultry protein compost continuing to compost afer bagging. The implicatons of this posibilty are ls than benefical. If cting is stil occurring post-bagging, then t physical and chemical properties of its contents are stil changing. Even furtr, this leads to bag-tobag difrencs, which w found in some of our studies. The somtims ?erratic? result came through in some of the studis. Whil ts cats a negative light, it should also be 51 noted that even though this happened, the Jungle Growth ? products sil performed as wl as or beter than their competiors in ts two studies. However, further evaluation of changes over tim of t poultry protein compost and the efcts on nutrition and other parametrs would be benefical for the cpany. Thereere case of whih a particular bag of substrae kild the plant it hosted within thre days of plnting, but subsequent bags of the sam formul or recipe resultd in some of the greates plant growth. Therfore, quality control should truly be evaluated within t cpany?s production line and within the individual components of the substrae mixes. Evaluation of Nem Percntages as an Organic Substrate Component: Again, as mntioned in the commercialy available organic substra comparison study, the poultry protein compost should be further eluatd, beause of the inconsistencis that sm to be apparent wn working with the substnc. Along the sam lines, the nee cake should be evaluated for its longevity. Azadirachtin, t cheial atributd to neem cake?s benefits, lvels should be looked at over time to determine if the cicl degrades with time, or in certain conditions that may be present in a bagged substrae, espeialy one that may continue composting after mixing and bagging. A beter choice of plant mterial could have been advantageous for this study. Salvi?s growth habit is not alwys esy to measure (as it did not grow verticaly, and ws quite britle) not only growth indics, but flowr count is more difiult. Beuse of this, it was dificult to ascertain neem?s efets on plant response. 52 Use of Nem Cake as an Organic Substrate Component: Whil ts experiment ss to have had fw obvious shortcomings, the initial experimental pln should have been st up factorily. Because the study took plce in a glas grenhouse, there becam a concrn after a fw dat-taking days that our own respiratory gas wre influencing the sampls collcted from the cmbers ? the gas chromatograph showed an unusualy high crbon dioxide value. Trefore, we moved olletion to outside of the greenhouse. This raied a concern, of sorts, in that the nvironmnt outsi was much more variant than in the grenhouse, so this may have cause som fluctuaion (i.c. temperature diferencs) in t substra respiration and subsquent gas sample values. Uing a covered aa could have benefitd greatly, tking out the varible of sun exposure and radiant het. Also, this particular project gives an incomplet answer to the question posed at the beginning. This study took a sondary look at nem?s efcts on urease activiy. We can asume a conclusion based on the collectd gas values, but a future study heavily steped in microbiology and soil pathology is neesary to determine the true interaction betwn neem and the bacteria producing ureas a wl as the urease itlf. 53 Apendix A Commercially Availabe Soiles Media Comparison Signficane to Industry: With ?gre-mindednes? steadily gaining popularity, the organic movement is beginning to hit the horticultural mrket. The efcts of eleven commercialy vailabl substraes on plant growth were evaluatd using four specis of plants. In experiment one, Jungl Growth ? organic substraes outperformed other commercil potting substraes and were only slightly infrior in perfanc to t best-ranking substrae, Miracl-Gro ? Moisture Control ? . In the second study, where diferent, the Jungl rowth substraes outperformed al otr substraes tted (though not staistcly greater than Sta-Gren ? Flowr and Vegetabl mix when usd on tomtoes and petunias). Introduction: The 1970?s horticulture industry had a ?breakthrough? in using pine bark as an invaluable resource, however present-day demnds for this commodity have also increasd and, with the growth of the horticulture sector, our industry is feling the presure to find some altrnatives to exclusive pine bark usage (Avent, 2003). While greenhouse plants arelmost exclusily produced in pet-substraes, 75-100%, by volum, of continer substraes in the eastern US are comprisd of pine bark (Lu et al., 2006). Future projections projct rising cost of pine bark cbid with les availbiity to the hortiulture industry (Lu et al., 2006). 54 Numerous organic and inorganic substances are being used to develop varying substras. Clan chip residual (the materil lft on the forest floor following timber harvest) cn be a relistc alternative to pine bark-exclusive substraes for ornaental plant production (Boyer et al., 2008). Fain et al. (2008) succesfully usd WholTre (whole pine tree shoots) in varying percentges a suitabl grenhouse mdia for growing marigolds and petunias. With additional frtilzer, Jckson and Wright (2007) were able to grow plants in 100% pulverized pine wood. Cotton gin compost was used as a viabl substrae component by Col t al. (2002). Spent tea grinds can be succsfully used in horticultural substraes (Wls, 2008). Poultry litr ws a suitble amndment for substraes omprisd primarily of either pine bark, WholeTre (creatd by chipping, then further milng entire pine trees ? log, limbs, needls and bark), or clen chip residual for growing plants in contairs with favorabl result and is a readily available source of nitrogen (Fin et al., 2008; Mrble et al., 2010; Mitchel and Donald, 1995). Parboiled fresh rice hulls can be used sucesfully as a substrae amndment comparabl to perlit (Evans and Gchukia, 2007). Using stras from peanut hulls, pine bark, peatmoss and combination thereof, Bilderback et al. (1982) reported succes in growing azles. In searching for an ecologicaly-friendly alternative to rockwool, Urrestarau et al. (2005) concluded that almond shels can be an efcti and benefical soiles mdia. Coconut coir can be a viable substrae for plnt production, however its succs depends on particle siz (Noguera et al., 2003). Pat-vermiculte substras had benefial result when growing chrysanthemums (Paul and Le, 1976). Mineral substraes have t potential to perform silarly to pet-basd mixes (Smith e al., 55 1995). Smith and Hal (1994) determined that a perlite-basd substrae can be comparable in mnagement and productiviy to peat-sd potting mixes. Whil potential atrnative substraes sm to abound, there a requirements that must be met of the mtril in order for it to be usable nd efective. Chang and Lin (2007) report that basic requirements for a succsful and benefial plnt-growing medium include: excelnt cheical resitance properties, light weight, inexpensivenes, abs of pest and disae and availbiity. Nkongolo and Caron (2006) noted that particle siz, specifly in pet and pine bark-based substraes, influences plant respons. Handrek (1983) reports the importanc of particl siz and advis that substrae formultors heed the ?fines? fraction of a substrae, especialy particle siz smalr than 0.5mm, as thi size controls the physical propertis of pine bark-basd substraes. Shrinkage is a physical property of organi soils (such as peat) that must be managed (Schw?rzel et al., 2002). Nemati e al. (2002) note that insufficent aration of rtifcl growing mdia is a comon proble in nursry and grenhous production. Azadirachta indic A. Juss, or the nem tree, is known as the ?vilage dispensary? in India and Southeastia, where its native (Biswa et al., 2002). This i due to the result of both scienc and tradition about the medicinal and agricultural uses nem provides. The nem tre is an evergreen tre belonging to the mahogany faily, Meliace, of which Melia azedarach Chinaberry is alo aember (Bisw et al., 2002). Stped in Indian tradition and lore, the neem tree continues to play roles in their traditions, medicine and agriculture and now the rest of the world is beginning to pay more atntion to neem, whih has been somewhat controversial to the traditionalist in Asia (Mrden, 1999). 56 Nem has been usd as a soil amendmnt in many studies in India. Nem sed residue provided a nitrogen value, aftr oil extraction, of 7% and at a rels rat fast enough to satisfy maize nutrition (Agbenin et al., 1999). Nem also enhanced plnt growth when incorporatd into soil at 2.5, 5.0 and 20 ppm, though 10 ppm decreased plant growth (Bhaskar and Charyulu, 2005). Likewise, Agyarko et al. (2006) reportd tha soil nutritional levels increased with increasing lvels of neem laves and poultry manure incorporation. Bhal and Dvi Prasd (2008) reportd higher vegetaive growth in plnts than reproductive growth (both showing higher growth than the control) when neem was incorporated into the soil. The objctives of this experiment wre to compare commercialy-vailable soiles potting media to determinehich is most efective in growing selctd annuals and tomatoes. Materials and Methods: Experimnt 1: Instaled on Septmber 16, 2009, seven commercialy available potting substraes wre usd, with each being its own treatnt and ech tretmnt containing twlve replicates. The seven substraes wre Jungle Growth ? products: Flower and Vegetabl, Profsional Mix and WatrWise ? ; and Scott?sts: Miracle- Gro ? Organic Choice ? , Miracle-Gro ? Moisture Control, Mirale-Gro ? Potting Mix and Scott?s Premium Potting Mix. Plant mterial used in the study included Petunia xhybrida and Tagetes patula which wre transplntd on Septmber 16, 2009, from 288- cell trays into Diln Products 6?Azalea pots (item #DIL60ATW) and grown in a double- layer poly greenhouse located at Auburn University?s Paterson Greenhouse Complx. Al plnts wre hand-wtred s needed and the trmost ws st a min/ax 57 temperatures of 65/78 o F. Pots were arranged in a randomized complet block design acording to replicate number. Dat wre taken at terminaton of the 8-wk study (56 days after planting, DAP) and included: growth indics (taken as [(height + width 1 + width 2 ) / 3] in centimters), SD-502 readings (mesure chlorophyll levels in the plants), flowr count, pH and electril conductiviy (EC). Al data wre analyzed using Waller-Duncan k-ratio t-tes. Also, it ismportant to note that no supplemntal frtilzer was ued for the duration of the study, only what ws premixed by the mnufcturer. Trminaton date was November 11, 2009. Experimnt 2: This study was a repeat of the first sudy with a few modificatons. On January 29, 2010, four species: Ptunia xhybrida ?Dreams Midnight?, Tagetes patul ?Durango Gold?, Dianthus plumarius ?Floral Lace? and Lycopsersicon lyopersicum ?Early Girl? were transplanted from 288-cel trays into Diln Products 6?Azalea pots (item #DIL60ATW). For dinthus and marigold, the same sven substraes (treatments) from the first experiment wre used. Ptuni and tomto recid four additional treaments; Scott?s ? Sding Soil and three Sta-Green ? products: Flower and Vegetable, Tre and Shrub and Al Purpose Potting Mix. Ech treatment was replicatd 12 tims. Again, al pots were placd in a randomized complet block design inside a greenhouse at the Paterson Grenhouse Complex. Plants wre hand-watered as need and the grenhous thermosta ws st a min/x temperatures of 65/78 o F . Dat tken was the same as t first experient and again analyzd using Waler-Duncn k-ratio t-tes. Also, no supplemntal frtilzr was applied during the 8-wk study. Terminaton date was Mrch 26, 2010. 58 Results: Experiment 1: Petunia: Petunias grown in MG Moisture Control ? had the greates growth (as indictd by growth indices, Table 7), followed by JG Profesional Mix and JG Flower & Vegetable. MG Organi Choic ? , MG Potting Mix and Scott?s ? Premium Potting Mix producd the last growth in petunias, though plants in JG WaterWis ? Mix were not diferent from Scott?s ? Premium Potting Mix. JG Flowr & Vgetabl and MG Potting Mix had the highest pH measurements, though they were not diferent from J Profesional Mix and JG WatrWis ? Mix (Tabl 7). MG Moisture Control ? had the lowst pH, but was not diferent fromG Organic Choice ? or Scott?s ? Premium Potting Mix. JG WterWis ? Mix had the highest EC masurent. MG Moisture Control ? had the next highest EC value, but was not difernt from J Flower & Vegetable. MG Organic Choice ? had the lowest EC value, but was not diferent from MG Potting Mix or Sott?s Premium Potting Mix. Petunias grown in Moisture Control ? had the highest SPAD measurement, but wre not diferent from JG Flower & Vegetable, JG Profesional Mix, or JG WaterWis ? Mix (Table 7). Ptunias grown in M Potting Mix had the lowest SPAD msurent, but wre not diferent from Scott?s ? Premium Potting Mix. Marigold: Marigolds grown in MG Moisture Control ? had the greates growth (Table 8). JG WterWis ? Mix produced the next greates growth, but ws not diferent from Scott?s ? Premium Potting Mix, JG Flower & Vgetbl, or JG Profesional Mix. Marigolds in MG Organic Choice ? producd the last growth. There was no staistl difrence in pH mesurements aong al of the substraes (ble 8). JG WterWis ? Mix had the highest EC value, but was diferent 59 from JG Flower & Vegetable, MG Moisture Control ? , or MG Potting Mix. Scott?s ? Premium Potting Mix had the lowst EC value, but was only diferent from JG WaterWis ? Mix. Marigolds grown in JG Profesional Mix had the highest SPAD value, but were not diferent from J WaterWis ? Mix, JG Flower & Vegetable, or MG Moisture Control ? (Table 8). Mrigolds grown in M Organic Choic ? had the lowest SPAD value. Experiment 2: Dianthus: Plants grown in JG Profesional Mix had the largest growth indics and wre not diferent from those in J Flowr & Vegetable Mix (Tble 9). Dianthus grown in MG Potting Mix had the next greates growth, but ws not diferent from J WaterWis ? Mix, or Scott?s ? Premium Potting Mix. The only difrenc in pH measureents aong the substraes wa betwen JG WaterWis ? Mix and MG Moisture Control ? (Table 9). MGoisture Control ? had the lowst pH, but was only diferent from JG WtrWis ? ix (with the highest pH). JG aterWis ? Mix had a higher EC value than al other mixes. Tre were no other difrencs in EC among the substraes. Plants grown in JG Profsional Mix had the highest flower count, followed by JG Flower & Vegetable (Table 9). MG Potting Mix had the next highest flr count, but was not difrent from J WtrWise ? Mix and Scott?s ? Premium Potting Mix. MG Moisture Control ? had the lowest flr count and was not diferent from MG Organic Choice ? . Dianthus grown in JG Profesional Mix had the highest SPAD value and were not diferent from those in J Flowr & Vgetable. T otr substrae yilded lowr SPAD 60 values than JG Profesional Mix and JG Flower & Vegetable and there were no diferencs among the otr treatments. Marigold: Plants grown in JG Profesional Mix had the greates growth and were not diferent from mrigoldsn in J Flowr & Vegetable (Tbl 10). Marigolds grown in MG Organic Choice ? had the least ? growth and wre not diferent from those n i Moisture Control. Tre wre no diferencs among plants grown in the other media. There were no diferencs among treatents in pH (Table 10). JG WaterWis ? Mix had t highest EC masureent. There wre no diferencs among the otr treaments in EC. arigolds grown in JG Flower & Vegetable had the highest flower count and were not diferent from those grown in JG Profsional Mix and JG WatrWis ? Mix (Tabl 10). Plants grown in M Organic Choice ? had the lowest fler count and were not diferent from mrigolds grown in MG Moisture Control ? or Scott?s ? Premium Potting Mix. lants grown in JG Flower and Vegetable and JG Profesional Mix had the greates SPAD values (no difrenc betwn the two treatmnts) (Table 10). Marigolds grown in MG Organic Choice ? had the lowest SPAD value and wre not difrent from those grown i Moisture Control ? . Tre wre no diferencs among the otr treamnts. Petunia: Plnts grown in JG Flower & Vegetable had the greates growth and were not difrent from those grown in J Profsional Mix or St-Gren ? Flower a 61 Vegetable (Table 11). Petunias grown in Scott?s ? Seding Soil had the least growth and wre not difrent from thos in MG Moisture Control. Sta-Green ? Al Purpose Potting Mix had the highest pH, but was not diferent from Scott?s Sding Soil, JG WatrWise ? Mix, Sta-Green ? Flower and Vgetabl, or Sta-Green ? Tree and Shrub (Tble 11). MG Organic Choic had the lest pH and was not difrent from JG Profesional Mix. J WaterWis ? Mix had t highest EC measureent. There wre no diferencs among the otr treatments in EC. Marigolds grown in Sta-Gren ? Flower and Vegetbl had the highest flower count (Tble 11). Plants grown in J Flr & Vtabl had t next hist flr count and wre not diferent from those in JG Profesional Mix or JG WaterWis ? Mix. Marigolds grown in MG Moisure Control ? had the lowst fler count and wre not diferent from those in Scott?s ? Seding Soil, MG Organic Choic ? , or Sta-Green ? Al Purpos Potting Mix. lants grown in JG Flower & Vegetable had the highest SPAD value and were not diferent from thosen in J Profsional Mix, Sta-Green ? Tree and Shrub, or Sta- Gren ? Flowr and Vgetable (Table 11). Marigolds grown in J WatrWise ? Mix had the lest SPAD value and wre not difrent from those in MG Organic Choic, MG Moisture Control ? , MG Potting Mix, Scott?s ? Premium Potting Mix or Sott?s ? Seding Soil. Tomato: Plants grown in JG Flower & Vegetable had the greates growth and were not diferent from those grown in Sta-ren ? Flowr and Vgetbl or JG Profsional Mix (Tabl 12). Tomatoes grown in MG Moisture Control ? had the least growth and were not diferent from those i Organic Choice ? . 62 Sta-Green ? Al Purpose Potting Mix had the highest pH, but was not diferent from Scott?s Sding Soil, Sta-Gren ? Flower and Vegetable, JG WterWis ? Mix, or Sta-Green ? Tree and Shrub (Tble 12). MG Organic Choic ? had the lowst pH and was not difrent from JG Profesional Mix, M Moisture Control, Scott?s ? Premium Potting Mix, M Potting Mix, or JG Flower & Vegetable. There were no diferencs among treatnts for EC. Flowr count was highest for tomatoes grown in JG Profesional Mix, and plants grown in JG Fler & Vegetble and St-Green ? Flower and Vgetable wre not diferent (Tabl 12). Tomatoes grown in M Moisture Control ? had the lowst fler count and were not diferent from those iG Organic Choice, MG Potting Mix, Sott?s ? Premium Potting Mix, Scott?s ? Sding Soil, or Sta-ren ? Al Purpose Potting Mix. Tomatoes grown in JG Flower & Vegetable had the highest fruit count and wre not diferent from those in J Profsional Mix, JG WaterWis ? Mix, or Sta-Green ? Flowr and Vegetabl. Fruit count was lowest for plnts grown in Moisture Control ? and tomtoes grown in MG Organic Choice ? , MG Potting Mix, Scott?s ? Premium Potting Mix, Scott?s ? Seding Soil, and Sta-ren Al Purpose Potting Mix were not diferent. Tomatoes grown in JG WaterWis ? Mix had the lowest SPAD value and were not diferent from MGoisture Control or MG Potting Mix (Tabl 12). Disregarding plants in J WaterWis ? Mix, there were no diferencs among al other treatments for highest SPAD values. 63 Discuion: Experiment 1: In referenc to growth indices, plants grown in Miracle-Gro ? Moisture Control ? outperformd al other treamnts in both the petunia and marigold studies. For the petunias and marigolds, Jungle Growth ? Flower & Vegetble and Profsional Mix produced the second largest plants. Jungl Growth ? WatrWis ? a Scott?s ? Premium Potting Mix, though, were not diferent from Jungle Growth Flower & Vegetable or Jungle Growth ? Profsional Mix in marigolds. For both petunia and marigold, plnts grown in Miracle-Gro ? oisture Control ? , Jungle Growth ? Flower & Vegetble, Jungle Growth ? Profsional Mix and Jungle Growth WatrWis Mix had the highest SPAD value and were not staistcly difrent. Experiment 2: For the purposes of this diussion, dianthus and marigolds are addresed in one sction and t petunias with the tomtoes in another, followed by overal conclusions based on the substraes usd: 7 products were usd with dianthus/marigold and 11 products wre usd with petunia/tomto. Dianthus/Marigold: In both case (dianthus and mrigold), Jungle Growth ? Flower & Vegetable and Profesional Mix outperformed the otr potting mixes in plant growth. Jungl Growth ? Profsilix producd t greates flower count in dinthus (with the second greates number being from Jungle Growth ? Flr & Vgetable). Marigold flowr count ws alo greates in the Jrowth products. It is alo importnt to note tha the EC for Jungl Growth ? WaterWis ? were significantly higher than the otr products in both the dianthus and mrigold cas. Petunia/Tomato: Again, Jungle Growth ? Flower & Vegetable and Profesional Mixs reultd in the greates growth for both petunia and tomto; howver, Sta-Gren ? 64 Flower and Vegetable is not diferent. Flower count for petunias w highest Sa-Green ? lr atbl followd scondly by the three Jungle Growth ? products (with no diferenc among the Jungle Growth ? treatments). Tomato flowr count was gretes aong Jungle Growth ? Flowr & Vegetbl, Jungle Growth ? Profesional Mix and Sta- Green ? Flowr and Vegetable (with non diferenc among the 3 treatmnts). Fruit count in tomatoes yielded the sm result as flowr count, with t exception of Jungle Growth ? WtrWis ? Mix being included among the three highest flower count treaments. Final EC result also show that Jungle Growth ? WaterWis ? Mix, in the case of the petunia study, was stistcaly greater tn al other mixes (there is no staistcl diferenc among any of the mixes in the tomto sudy). Overall: Jungle Growth ? Flower & Vegetable and Profesional Mix, as wel as Sta-Gren ? Flowr and Vgetabl are most beneficl for plant growth, for the four species tted in this experiment. Growth indies, flower count and fruit count (in the as of tomatoes) reveal that Jungl Gth ? outperforms al other treatments tted, other than St-Green ? Flower and Vegetable when used, for al four specis of plants. Whil Experimnt 1 indicatd that Miracl-Gro ? Moisture Control ? produced the greates plant performance, followed by the Jungle Growth products, the sam result did not occur in Experimnt 2. Miracl-Gro ? Moisture Control ? was not a competior in this cae. It should be noted that one component of the Jungle Growth ? products that was not part of any other treatmnt is neem cake powder. Isolating this diferenc drew us to pursue furtr reserch on nee cakeder?s activiy within the soil and its subsequent efct on plants. 65 Literatue Cited: Agbenin, J.O., E.B. Agbaji, I. Suleiman, and A.S. Agbaji. 1999. 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