Effect of Growing Factors on Productivity and Quality of Lemon Catmint, Lemon Balm and Sage under Soilless Greenhouse Production: I.

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1 Mediinl nd Aromti Plnt Siene nd Biotehnology 2 Glol Siene Books Effet of Growing Ftors on Produtivity nd Qulity of Lemon Ctmint, Lemon Blm nd Sge under Soilless Greenhouse Prodution: I. Drought Stress Artur Mnukyn Center of Greenhouses nd Lortories Dürnst, Center of Life Sienes Weihenstephn, Tehnil University of Munih, Dürnst 4, 8535 Freising-Weihenstephn, Germny Corresponding uthor: * mnukyn@wzw.tum.de ABSTRACT The formtion of iotive onstituents of plnts depends on the tul environmentl nd on growing ftors. Controlled environment tehnology ensures the pplition of speifi stresses, suh s drought stress, tht n optimize the prodution of seondry plnt metolites y induing nturl iohemil hnges in plnts. The impt of drought stress on herl yield, ontent nd omposition of essentil oils nd polyphenols, s well s on ntioxidtive pity of polyphenol-rih extrts of lemon tmint (Nepet tri L. f. itriodor), lemon lm (Meliss offiinlis L.) nd sge (Slvi offiinlis L.) ws studied under soilless greenhouse ultivtion. It ws found tht the tested sustrte moistures (5, 25, 25 hp) hd onsiderle effet on herl yield of lemon tmint, lemon lm nd sge with mximum rw mediinl mteril t 5 hp. Drought stress influened the essentil oil ontent of lemon tmint nd lemon lm, ut not of sge, with 25 hp resulting higher mount of essentil oils. 5 hp provided high yield of essentil oil for ll three speies. The essentil oil omposition of studied hers ws minly ffeted y tested sustrte moistures. The influene of drought stress ws lso signifint for the ontent of polyphenols in lemon lm nd sge, ut not in lemon tmint. High drought stress (25 hp) ffeted positively polyphenoli ontent in lemon lm, while in sge polyphenols rehed mximum t 25 hp. HPLC nlysis of polyphenols showed differenes in sensitivity of min polyphenols in studied hers to drought stress ftor. Polyphenol-rih extrts of lemon tmint nd lemon lm showed no differenes in ntioxidtive pity y the ABTS system, while for sge there were smll differenes. Keywords: iotive ompounds, hydroponis, Meliss offiinlis, Nepet tri f. itriodor, Slvi offiinlis, sustrte moisture INTRODUCTION Soilless greenhouse prodution of mediinl nd romti plnts is perspetive nd eonomil lterntive for high produtivity, superior qulity with eologil sound nd ensures tht the plnts free from ioti nd ioti ontmintions with onsistent iohemil profiles. For these purposes n dpted tehnology, inluding informtion on effets of stress ftors, suh s drought stress, is required. Growing nd environmentl ftors ply mjor role in mngement of herl yield nd qulity providing mediinl plnt mteril with the use of nturl tools (Frnz 983; Hornok 986; Bettry et l. 992; Mnukyn 24, 24; Zoyed et l. 25; Khlid 26; Mnukyn et l. 26; Belitz et l. 27; Kooheki et l. 28; Rzmjoo et l. 28; Frhni et l. 29; Sid-Al Ahl et l. 29; Bghlin et l. 2; Li et l. 2). The optiml tehnology tht will llow produing high qulity plnt mteril with minimum mounts nturl resoures (e.g. wter) is environmentlly sound nd highly neessry for modern prodution proess (Shnitzler et l. 23). As for plnt prodution in generl, it is likewise true for growing of mediinl nd romti plnts. The im of this study is to investigte the effet of drought stress on herl yield, mount nd omposition of essentil oils (EOs) nd flvonoids, s well s on ntioxidtive pity of flvonoid-rih extrts of some vlule mediinl plnts under soilless greenhouse onditions. Suh informtion provides sis to modify nd optimise the qulity of mediinl nd romti plnts, using onventionl plnt reeding omined with mnipultion of gronomi prties. MATERIALS AND METHODS Plnt mteril nd experimentl design The experiments were onduted t the Center of Greenhouses nd Lortories Dürnst of the Center of Life Sienes Weihenstephn, Tehnil University of Munih. Mediinl nd romti plnts to study were lemon tmint (Nepet tri L. f. itriodor), lemon lm (Meliss offiinlis L.) nd sge (Slvi offiinlis L.) - (ll three speies elong to the Lmiee fmily). Cultivtion ws strted y sowing seeds, derived from hydroponilly grown plnts, in ommon pet ( Florton 3 ) in green- Fig. Mediinl nd romti plnts under soilless greenhouse onditions. Reeived: 2 My, 2. Aepted: 3 June, 2. Originl Reserh Pper

2 Mediinl nd Aromti Plnt Siene nd Biotehnology 5 (2), Glol Siene Books house. Seeds were strtified y pplition of 2 g/l KNO 3 solution (Merk) nd C for one week. Priking ws done one month lter into ell trys with ommon pet sustrte ( Florton 2 ). The experiment strted nother month lter with trnsplnting of the seedlings into 3 l (lemon tmint nd sge) nd 5 l (lemon lm) pots. The plnts were ultivted in utomtilly ontrolled greenhouse (Fig. ). For ll speies unomposted wood fire Tores speil (Ts) ws used s sustrte, whih hd ph 5.9 nd ontined.8 g/l wter-solule slts (233 mg/l N, 9 mg/l P 2 O 5 nd 9 mg/l K 2 O). For eh speies 2 e-nd-flood tles were used with 3.3 m 2 surfe re for eh with 9 plnts per m 2 (3 plnts per tretment with 4 repetitions). The nutrient solution ws mde up from 7 g Flory sis (P-43; K-22; Mg-2; B-.; Cu-.5; Fe-; Mn-.2; Mo-.3; Zn-.25 mg/l) in ddition to 5 mg/l N (omposed of 2.4 g mmonium nitrte (Merk) nd 48.2 g lium nitrte (Merk) per l of wter) with ph of nd eletril ondutivity (EC) of ms/m. The irrigtion of plnts ws performed vi tensio swith sensors providing sustrte moisture of 5, 25, 25 hp. Plnts were hrvested t week 8 fter trnsplnting. The riterion for lemon tmint hrvest ws the time of flower emergene; the other two speies were hrvested lmost t the sme time hving minimum 8 (for lemon lm) nd 5 g/plnt (for sge) fresh iomss. The fresh mediinl mteril ws dried t 35 C for 5-7 dys. Anlyses EO from fresh herl mteril of seleted mediinl nd romti plnts were extrted y stem distilltion nd pentne (Crl Roth) extrtion for h (SDE). The extrts were dried over sodium sulfte (Merk) nd onentrted under strem of nitrogen. The mount of essentil oil ws determined grvimetrilly. The omposition of the essentil oil ws nlyzed y GC nd GC-MS. The oils were diluted in etone (Crl Roth) (split : 4) with seprtion of the ompounds y GC (Fisons Instruments Meg 536, Itly) on Supelo-Wx pillry olumn (6 m, i.d. =.32 mm,.25 m film thikness) with helium s rrier gs (.8 ml/min) nd temperture progrm: 5 C (3 min), C/min, 2 C (2 min), 2 C/min, 55 C ( min), 8 C/min, 24 C. Identifition of EO min ompounds ws performed with GC-MS system (HP 589 Series II/HP 597 A, HEWLETT PACKARD) on the sme olumn nd with the sme temperture progrm y eletron impt ioniztion t 7 ev. Mss spetr were evluted y omprison of retention times nd mss spetr (Wiley 99) nd with n own terpenoid mss spetr dtse. Polyphenols were extrted from.4 g dry mteril (powder) with 8% MeOH (Crl Roth) nd seprted y HPLC. Extrted polyphenols were resolved in 5 μl CH 3 CN (Merk) nd 5 μl EtOH (Merk), injetion volume ws μl. The nlytil HPLC ws rried out on dionex system (pump P58, utosmpler Gin 5) using 25 mm x 4,6 mm RP-8 olumn (phenomenex Hydro-RP) with gurd olumn. A grdient sequene using (A) wter, etonitrile nd eti id (Merk) (97: 2: ) nd (B) etonitrile in the following proportions: % B ( min), 5% B (5 min), 95% B ( min), % B (4 min) t flow rte of ml/min. Detetion nd quntifition were performed using Diode Arry detetor (Dionex UVD 34S) nd hromeleon (Dionex) softwre. Folin-Ciolteu test (Merk) pplied to determinte the totl ontent of polyphenolis in plnt mteril (Singleton et l. 998). The ABTS-system ws used to evlute the ntioxidtive pity of polyphenol-rih extrts (Arno et l. 999; Re et l. 999). The ility to redue the peroxidse (myelogloin/h 2 O 2 )-generted ABTS + [2,2'-zino-is(3-ethylenzthizoline-6-sulfoni id) rdil tion] hs een used to rnk the ntioxidnt tivity of vrious gents inluding dietry flvonoids nd hlones (Chn et l. 23). In this system, myogloin (Sigm) nd H 2 O 2 (Merk) oxidise ABTS (Merk) to the green ABTS + rdil tion. 2,2'-zino-is(3-ethylenzthizoline-6-sulfoni id) rdil tion (ABTS +) is stle hromophore whih sors strongly t 734 nm. This retion n e followed photometrilly. The lower rte of ABTS + formtion indites reduing or Fe-helting properties of the extrt. The nlyses were performed with 3-4 replitions. The sttistil nlyses were performed ording to Dospekhov (985). RESULTS The herl yield of studied speies ws ffeted y different sustrte moistures. High drought stress influened negtively the produtivity of hers. At 25 hp, lemon tmint, lemon lm nd sge produed respetively 42.4, 25.4 nd 5.9% less totl rw mediinl mteril thn t 5 hp. In se of lemon lm nd sge high drought stress of 25 hp provided higher perent of dry weight (Tle ). Tested sustrte moistures influened the EO ontent of lemon tmint nd lemon lm (Fig. 2). High drought stress (25 hp) indued higher mount of EO in lemon tmint nd lemon lm (.5 nd.68%, respetively), while in the se of sge there ws no signifint differene etween the tretments. Clultions of EO yield provided sttistil differenes for ll three speies. 5 hp tretment ws est for lemon tmint, lemon lm nd sge (.363,.87 nd.98 ml/plnt, respetively) (Fig. 3) due to the high fresh herl iomss per plnt (see lso Tle ). Lemon tmint EO omposition ws ffeted y the different sustrte moistures under soilless greenhouse ondi oil ontent [%],8,6,4,2 5 hp 25 hp 25 hp 5 hp 25 hp 25 hp 5 hp 25 hp 25 hp Lemon tmint Lemon lm Sge Fig. 2 Influene of sustrte moisture on essentil oil ontent of hers. Vlues represent men ± stndrd devition (SD). Sttistis re mde within the sme speies. 2

3 Drought stress nd soilless ulture of hers. Artur Mnukyn Tle Influene of sustrte moisture on herl yield. Speies Sustrte Fresh mediinl Rw mediinl mteril moisture, hp mteril, g/plnt g/plnt % (dry weight) Lemon tmint LSD 5* Lemon lm LSD 5* Sge LSD 5* * Within the sme speies Tle 2 Influene of sustrte moisture on lemon tmint essentil oil omposition. Compound* Sustrte moisture, hp Content in essentil oil, % is-oimene.4 **.3.4 is-3-hexenl trns-oimene...2 is-3-hexenyl ette methyl-5-hepten-2-on trns-3-hexen--ol is-3-hexen--ol trns-2-hexen--ol itronelll linlool..9. -ryophyllene..8. itronellyl ette humulene nerl neryl ette gernil gernyl ette.7.8. itronellol nerol gerniol ionone..2. phytol * Only identified ompounds re presented ** Sttistil nlyses re mde within the sme ompound oil totl yield [ml/plnt],4,35,3,25,2 5,5 5 hp 25 hp 25 hp 5 hp 25 hp 25 hp 5 hp 25 hp 25 hp Lemon tmint Lemon lm Sge Fig. 3 Influene of sustrte moisture on essentil oil yield of hers. Vlues represent men ± stndrd devition (SD). Sttistis re mde within the sme speies. Tle 3 Influene of sustrte moisture on lemon lm essentil oil omposition. Compound* Sustrte moisture, hp Content in essentil oil, % -myrene.6 **.7.8 is-oimene is-3-hexenl.2..2 trns-oimene methyl-5-hepten-2-on trns-3-hexen--ol trns-2-hexen--ol oten-3-ol itronelll linlool not identified not identified ryophyllene humulene nerl neryl ette gernil gernyl ette itronellol nerol gerniol is-9-otdeenoi id methyl ester not identified not identified * Only identified nd not identified min sustnes of out or > % re presented ** Sttistil nlyses re mde within the sme ompound Tle 4 Influene of sustrte moisture on sge essentil oil omposition. Compound* Sustrte moisture, hp Content in essentil oil, % -pinene.58 ** mphene pinene sinen myrene terpinene limonene ,8-ineole is-oimene.8.2. is-3-hexenl terpinene p-ymol terpinolene trns-3-hexen--ol trns-2-hexen--ol thujone thujone mphor ornyl ette ryophyllene humulene orneol myrtenol viridiflorol mnool * Only identified ompounds re presented ** Sttistil nlyses re mde within the sme ompound tions (Tle 2). Min ompounds of the EO were gerniol ( %), nerol ( %), gernil ( %), itronellol ( %) nd nerl ( %). The identified 22 ompounds represented pproximtely 97, 98 nd 98% of the totl oil t 5, 25, 25 hp sustrte moisture, respetively. Drought stress hd onsiderle impt on the quntity of some minor ompounds in lemon tmint EO, suh s trns-oimene, is-3-hexenyl ette, 6-methyl-5-hepten-2-on, trns-3-hexen--ol, is-3- hexen--ol, trns-2-hexen--ol, linlool, -ryophyllene, neryl ette, gernyl ette, -ionone nd phytol. High drought stress of 25 hp provided high ontent of trnsoimen, is-3-hexenyl ette, neryl ette, gernyl ette nd phytol in lemon tmint EO, while the ontent of 6-2

4 Mediinl nd Aromti Plnt Siene nd Biotehnology 5 (2), Glol Siene Books Tle 5 Influene of sustrte moisture on polyphenol ontent of hers. Speies Sustrte moisture, hp Glli id equivlents- GAE [mm] Lemon tmint 5.65 ±, ±, ±, LSD 5* Lemon lm 5.8 ±, ±, ±,5 LSD 5* Sge 5.63 ±, ±, ±,4 LSD 5* * Within the sme speies pek / internl stndrd 6 4 2,8,6,4,2 5 hp 25 hp 25 hp 5 hp 25 hp 25 hp 5 hp 25 hp 25 hp Pek (ffei id) Pek 2 (rosmrini id) Pek 3 (p-oumri id) Fig. 4 Influene of sustrte moisture on quntity of mjor polyphenols in lemon tmint. Vlues represent men ± stndrd devition (SD). Sttistis re mde within the sme ompound. pek / internl stndrd hp 25 hp 25 hp 5 hp 25 hp 25 hp 5 hp 25 hp 25 hp Pek (ffei id) Pek 2 (rosmrini id) Pek 3 (melitri id) Fig. 5 Influene of sustrte moisture on quntity of mjor polyphenols in lemon lm. Vlues represent men ± stndrd devition (SD). Sttistis re mde within the sme ompound. pek / internl stndrd hp 25 hp 25 hp 5 hp 25 hp 25 hp 5 hp 25 hp 25 hp Pek (ffei id) Pek 2 (rosmrini id) Pek 3 (melitri id) Fig. 6 Influene of sustrte moisture on quntity of mjor polyphenols in sge. Vlues represent men ± stndrd devition (SD). Sttistis re mde within the sme ompound. sorption 734 nm,9 Control,8 5 hp 25 hp,7 25 hp,6,5,4,3, retion time [min] Fig. 7 Influene of sustrte moisture on ntioxidtive pity of polyphenol-rih extrts of lemon tmint. sorption 734 nm,9 Control,8 5 hp 25 hp,7 25 hp,6,5,4,3, retion time [min] Fig. 8 Influene of sustrte moisture on ntioxidtive pity of polyphenol-rih extrts of lemon lm. sorption 734 nm,9 Control 5 hp,8 25 hp,7 25 hp,6,5,4,3, retion time [min] Fig. 9 Influene of sustrte moisture on ntioxidtive pity of polyphenol-rih extrts of sge. methyl-5-hepten-2-on nd is-3-hexen--ol deresed. The ontent of trns-3-hexen--ol, linlool, -ryophyllene nd -ionone rehed mximum t 25 hp nd fter tht deresed gin. At the sme time, for the ontent of lemon tmint EO min ompounds, drought stress ftor hd no signifint effet. Lemon lm EO onsisted minly of gernil ( %), nerl ( %) nd gerniol ( %) under soilless greenhouse prodution (Tle 3). 2 ompounds representing out 89, 95 nd 97% of the totl oil (t 5, 25, 25 hp, respetively) were identified. The ontent of ll these ompounds, exept for -oten-3-ol nd nerol, depended on moisture level in used wood fire sustrte. From the min ompounds of lemon lm EO, the ontents of gernil nd nerl peked up with inresing of drought stress, while gerniol rehed mximum when plnts were grown t 5 hp. For sge, the 25 identified ompounds of EO represented pproximtely 97% of the oil omposition t 5, 25, 25 hp. Among these, -thujone ( %), mphor ( %), -thujone ( %),,8-ineole ( %) nd viridiflorol ( %) were the mjor onstituents under soilless greenhouse onditions (Tle 4). Drought stress hd onsiderle influene on the ontent of most identified ompounds in sge EO, suh s -pinene, 22

5 Drought stress nd soilless ulture of hers. Artur Mnukyn mphene,,8-ineole, is-oimen, is-3-hexenl, p-ymol, terpinolene, trns-3-hexen--ol, trns-2-hexen--ol, -thujone, -thujone, mphor, ornyl ette, -ryophyllene, -humulene, orneol, myrtenol, viridiflorol nd mnool. From the min ompounds the ontent of -thujone nd viridiflorol ws higher t 5 nd 25 hp, while high drought stress of 25 hp resulted mximl mount of thujone. The ontent of other two min ompounds (,8- ineole nd mphor) ws minimum t 25 hp. The influene of drought stress ws lso essentil for the ontent of polyphenols in lemon lm nd sge, ut not in lemon tmint under soilless greenhouse onditions (Tle 5). Lemon lm nd sge ontined more polyphenoli ompounds thn lemon tmint. High drought stress of 25 hp ws effetive for iosynthesis of polyphenols in lemon lm; while for sge we hve reorded higher ontent of polyphenols t 25 hp. HPLC nlysis of min polyphenols provided differenes in sensitivity of min polyphenols in seleted hers to drought stress ftor. In lemon tmint, ll three min polyphenoli ompounds (ffei, rosmrini nd p-oumri ids) showed high results t 5 hp (Fig. 4). In lemon lm, min pek 2 (rosmrini id) rehed mximum t high drought stress (25 hp), while pek (ffei id) topped t 25 hp nd pek 3 (melitri id) t 5 hp (Fig. 5). In sge, min pek 2 (rosmrini id) nd pek (ffei id) rehed mximum results t 5 hp, while pek 3 (melitri id) in se of high drought stress tretment (Fig. 6). At the sme time we hve reorded tht onentrtion of rosmrini id in lemon lm nd sge ws. times higher thn in lemon tmint. Polyphenol-rih extrts of lemon lm nd sge hd higher ntioxidtive pity in the ABTS-system nd were tested in -fold lower onentrtions (: 2) (Figs. 8-9) thn those of lemon tmint (: 2) (Fig. 7). Polyphenolrih extrts of lemon tmint grown under different sustrte moisture onditions showed no essentil differenes in ntioxidtive pity (Fig. 7). In se of lemon lm there were no signifint differenes s well nd the shpes of the urves were lmost synhroni (Fig. 8). Polyphenolrih extrts of sge showed smll differenes in ntioxidtive pity y the ABTS model system, prtiulrly 25 hp tretment with high ontent of polyphenols (see lso Tle 5) produed higher ntioxidtive pity (Fig. 9). DISCUSSION The intensive use of wter nd fertiliser in modern hortiulture reflets need for developing new nd more environmentl suitle prodution methods to redue the usge of wter nd fertilizer. At the sme time the improvement of inner qulity of useful speies is highly neessry. In this study we hve foused on drought s single stress ftor. Our results showed tht omprtively high drought stress (25 hp) influened negtively the produtivity of studied hers s wited. At the sme time some differenes noted in onnetion to the influene of drought stress ftor on herl yield, s there were stronger differenes in se of sge nd lemon tmint. High drought stress ffeted positively EO ontent in lemon tmint nd lemon lm, ut this dvntge ws not enough for high yield of EO, euse of low fresh herl yield in tht tretment. Drought stress minly ffeted the EO omposition of studied hers under soilless greenhouse onditions. For polyphenoli ontent nd omposition of seleted mediinl nd romti plnts we hve seen essentil influenes s well, while in se ntioxidtive pity of polyphenol-rih extrts we hve reorded omprtively less ritil impts. This my e explined y speifi inhiition potentils of some polyphenols in the extrts of seleted hers. Severl uthors hve pulished dt on the effets of irrigtion regime, sustrte moisture nd drought stress on produtivity nd qulity of different mediinl nd romti plnts. The impt of ove-mentioned ftors on iosynthesis of some phrmeutil ompounds in mediinl plnts ws desried oth s positive nd negtive or neutrl depending on ertin iotive ompound nd speies. One reserh study investigted the effets of ute periods of drought stress nd plnt ge on dry weight, nd lkmide nd phenoli id ontent in purple oneflower roots [Ehine purpure (L.) Moenh] nd found tht the totl lkmide, inluding the tetrenoi id isomers, nd hlorogeni id onentrtions from fll-hrvested roots were lrgely unffeted y drought stress, regrdless of t whih stge the stress ourred (Gry et l. 23). On the other hnd, mny other reserhes showed ertin effet of drought stress on iotive onstitution of some hers. For instne, in n experiment, where Niotin glu plnts were sujeted to different irrigtion levels, it ws found tht inresed irrigtion frequeny effet negtively the lkloids perentge in the leves, t the sme time the highest rutin perentge ws found with the shorter irrigtion intervls (Sleh et l. 978). At the sme time, drought stress effets derese of the indole lkloid in Cthrnthus roseus (Frishkneht et l. 987) nd influenes negtively on the ontent of digitoxigenin in woolly foxglove Digitlis lnt, wheres the other rdenolides, inluding digoxin, were less ffeted (Stuhlfuth et l. 987). It ws lso shown tht in dedly nightshde (Atrop elldonn L.) there is orreltion etween the yield of tropne lkloids (hyosymine nd sopolmine) nd irrigtion regime, totl nitrogen supply. The mximl yield of tropne lkloids ws hieved when plnts grown under low ville soil wter (35%) nd low nitrogen supply onditions. By ontrst, the mximl ontent of lkloids ws hieved with 95% depletion of ville soil wter ompnied with high nitrogen supply (Bri evi et l. 22). A numer of phytohemils in ultivted St. John s wort, inluding hyperiin, pseudoohyperiin, hlorogeni id, rutin, hyperoside, isoqueritrin, queritrin, nd queretin, re signifintly inresed eing influened y ute drought stress. Inreses rnged from 5 to 36% (hyperoside nd rutin, respetively). Conversely, the onentrtions of hyperforin nd dhyperforin were deresed y n verge of % in drought-stressed plnts s ompred to well-wtered plnts (Gry et l. 23). The derese in soil field pity from to 4% redued EO yield of drought-tolernt nd non-droughttolernt lones of Tgetes minut under greenhouse onditions. The drought signifintly ltered the ontent of some oil omponents (Mohmed et l. 22). Wter stress influenes the EO s well s its onstituents of etelvine (Piper etle L.), effets eing depended on mgnitude of drought stress (Chtterjee 999). It resulted in signifint redutions in hlorophyll, rotenoids, Fe, Mn, Zn, nd EO yield of Jpnese mint (Menth rvensis L. v. MS 77) (Misr et l. 2). Drought nd old stress tretments used inresed in levels of (-)-epitehin nd hyperoside in Crtegus levigt nd Crtegus monogyn (hwthorn) plnts. Suh tretments lso enhned the ntioxidnt pity of the polyphenoli extrts (Kirkosyn et l. 23). Meliss offiinlis L. ws found highly tolernt ginst wter stress. Dry yield under wter defiit (, 2.5, 25., 37.5 nd 5.%) vried from 3.5 to 9.2 g/plnt. Redution in yield ws not sttistilly signifint till 25% wter defiieny, while the EO rtio inresed with eh inrese in wter defiieny (Ozturk et l. 24). In one study Slvi offiinlis L. ws sujeted to five different wtering levels (, 75, 5, 25 nd % of ETo) under limti onditions of extremely high temperture. It ws found there were not signifint differenes mong the monitored wter rtes with regrd to the existing reltion etween the units of dry mtter nd the EO yields. Wter rtes ffeted the omposition of Slvi offiinlis EO, prtiulrly the onentrtions of -pinene, mphene, ineole, - nd -thujone, ornyl ette nd -humulene (Corell et l. 29). In other study the endogenous levels of diterpene rnosi id nd -toopherol were mesured in drought-reovery yle in leves of sge (Slvi offiinlis L. sus. offiinlis), drought-suseptile speies, growing in Mediterrnen field onditions. As the drought progressed, the rnosi id 23

6 Mediinl nd Aromti Plnt Siene nd Biotehnology 5 (2), Glol Siene Books mounts in the leves deresed, giving rise to inresed levels of its oxidtion produts, rosmnol nd isorosmnol. At the sme time, -toopherol levels deresed progressively with drought (Munné-Bosh et l. 2). Drought stress lso deresed signifintly the folir ftty id ontent in Slvi offiinlis eril prts. Besides, moderte wter defiit inresed the EO yield nd the ontent of min voltile onstituents mphor, -thujone nd,8-ineole (Bettie et l. 29). The ojetive of nother study ws to investigte the effet of drought stress on herl growth nd omposition of tive onstituents in Slvi miltiorrhiz Bunge. The results showed tht exept rosmrini id the ontent of other tive onstituents inresed under wterstress onditions. Wter stress signifintly inresed slvinoli id B yield nd deresed tht of tnshinone IIA (Liu et l. 2). Following experiment investigted the influene of soil wter stress on plnt height, fresh nd dry weight nd EO ontent of Irnin Sturej hortensis L. The results showed tht the umultion of EO inresed signifintly under severe wter stress t the flowering stge, when the men lef wter potentil deresed from.5 to.6 MP. This tretment ffeted the quntity of the EO more thn moderte wter stress during the vegettive nd flowering stges. From the min oil onstituents the mount of rvrol inresed under moderte stress, while terpinene ontent deresed under moderte nd severe wter stress tretments (Bher et l. 22). It ws found tht the plnt growth of two lemongrsses [Cymopogon nrdus. (L.) Rendle vr. onfertiflorus (Steud.) Bor. nd Cymopogon pendulus (Steud.) Wts.] ws redued onsiderly y drought stress whilst the level of EO ws mintined or enhned. Gerniol nd itrl, s the mjor EO onstituents, inresed sustntilly in oth speies (Singh-Shgwn et l. 994). Drought stress ffeted growth nd EO metolism in itronell jv (Cymopogon winterinus) ultivrs. In generl, plnt growth ws redued onsiderly while the level of EO ws enhned under wter stress. The mjor oil onstituent itronelll deresed signifintly in one tested ultivr, while itronellol ontent, sisi id nd IAA inresed in oth the ultivrs under drought stress onditions (Shih et l. 2). Drought stress enhned lkloid jmliine prodution in Cthrnthus roseus (L.) G. Don plnts (Adul Jleel et l. 28). In other experiment with two vrieties (rose nd l) of Cthrnthus roseus, the vritions in ntioxidnt potentils (superoxide dismutse, sorte peroxidse, tlse, peroxidse nd polyphenol oxidse) nd indole lkloid ontent were studied, exposed to wter defiit stress. The results showed tht the ntioxidnt onentrtions nd tivities of ntioxidnt enzymes were high under wter defiit stress in ll prts of the plnts. Indole lkloid ontent ws high in the roots of rose vriety in response to drought stress when ompred to l vriety (Adul Jleel et l. 28). In other reserh the EO ontent nd hemil omposition of Germn hmomile (Mtriri hmomill L.) were determined t different irrigtion regimes (irrigtion t, 85, 7 nd 55% of field pity). Results indited tht the highest mount of EO perent, yield of dry flower nd essentil oil per pot were otined from irrigtion t 85% of the field pity nd lowest mount of EO perent were otined when the plnts irrigted with nd 55% of field pity (Pirzd et l. 26). Wter stress hs signifint effets on morphologil nd iohemil hrteristis of purple sil (Oimum silium). As the soil wter ontent deresed, herl yield nd lef hlorophyll ontents (, nd totl hlorophyll) deresed, ut the mounts of nthoynin nd proline inresed (Moeini Alishh et l. 26). Wter stress (wterlogging nd drought) inresed the levels of etulini id nd phenoli ompounds (queretin, rutin,,5-dihydroxyxnthone, isouliginosin B) in Hyperium rsiliense Choisy plnt (Nif de Areu et l. 25). The ojetive of nother study ws to determine the ontents of seondry metoli sustnes suh s resvertrol, SOD, phenoli ompounds nd free mino ids in Rehmnni glutinos under temperture nd wter stress. Of the 6 individul phenoli ompounds, myrietin showed the highest onentrtions in the wter defiieny (.8 MP) nd ontrol (.4 MP) nd low temperture (5 C) tretment. The 2 free mino ids in R. glutinos deresed t high temperture nd with wter defiieny (Chung et l. 26). In the following study, the hnges in the lkloid ontent (ererine, jtrorrhizine nd plmtine) in Amur orktree (Phellodendron murense) under different wter onditions (mild drought, severe drought, wterlogging nd ontrol) were disussed. It ws shown tht mild drought ws generlly enefiil to the synthesis nd umultion of the ove-mentioned lkloid ontents. The three lkloid ontents did not show gret hnges under severe drought, wheres those ontents hd signifintly redued under wterlogging ompred with ontrols (Xi et l. 27). An experiment ws rried out to determine the ssoition of te polyphenols with wter stress nd their suitility s inditors for drought tolerne. The experiment ws onduted in rin-out shelter, nd onsisted of six te lones nd four levels of soil wter ontents (38, 3, 22, nd 4% v/v). The results indite tht delining soil wter ontent redued oth growth nd ontent of polyphenols in te (Cheruiyot et l. 27). Wter stress inresed the yield of EO from the leves of plinlefed nd urly-lefed ultivrs of prsley, ut not turniprooted ultivr. Wter stress lso used hnges in the onstituents of EO (priniplly,3,8-p-menthtriene, myristiin, terpinolene + p-ymenene), ut these hnges vried etween ultivrs (Petropoulos et l. 28). Wter defiit indued signifint redution in growth prmeters nd ftty id ontent nd n inrese in the EO oil ompounds (rvone nd limonene) in rwy (Crum rvi L.) (Lrii et l. 29). CONCLUSION It n e onluded tht in soilless greenhouse prodution the produtivity nd qulity of lemon tmint (Nepet tri L. f. itriodor), lemon lm (Meliss offiinlis L.) nd sge (Slvi offiinlis L.) minly depended on drought stress. Herl yield, mount nd omposition of essentil oils nd polyphenols signifintly depended on sustrte moisture, while ntioxidtive pity of polyphenol-rih extrts ws ffeted to lesser extent. ACKNOWLEDGEMENTS This work ws prtly supported y the Interntionl Centre for Sientifi Culture (ICSC) - World Lortory (Lusnne, Switzerlnd). REFERENCES Adul Jleel C, Snkr B, Murli PV, Gomthinygm M, Lkshmnn GMA, Pnneerselvm R (28) Wter defiit stress effets on retive oxygen metolism in Cthrnthus roseus; impts on jmliine umultion. 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