POJ 5(3):271-278 (212) ISSN:1836-3644 Slinity effects on mcro nd micronutrients uptke in medicinl plnt King of Bitters (Androgrphis pnicult Nees.) Dryush Tlei 1, 2,*, Mihdzr Adul Kdir 3, Mohd Khnif Yusop 4, Alirez Vldini 3 nd Mohd Pud Adullh 1 1 Deprtment of Cell nd Moleculr Biology, Fculty of Biotechnology & Biomoleculr, Universiti Putr Mlysi, 434 UPM, Serdng, Selngor, Mlysi 2 Medicinl Plnt Reserch Centre, Shhed University, Tehrn, Irn 3 Deprtment of Agriculture Technology, Fculty of Agriculture, Universiti Putr Mlysi, 434 UPM, Serdng, Selngor, Mlysi 4 Deprtment of Lnd Mngement, Fculty of Agriculture, Universiti Putr Mlysi, 434 UPM, Serdng, Selngor, Mlysi *Corresponding uthor: khnif@gri.upm.edu.my nd Tlei1348@gmil.com Astrct Plnts species require mcro nd micro nutrients to complete the growth cycles such s photosynthesis, enzyme ctivities nd secondry metolite production. The ojective of the present study ws to investigte the effects of slinity on iomss s well s mcro nd micro nutrients uptke in the medicinl plnt of Androgrphis pnicult (AP). In this regrd, split plot experiment ws crried out sed on Rndomized Complete Block Design (RCBD) with two fctors; five slinity levels (control, 4, 8, 12 nd 16 dsm - 1 ) in min plots nd 12 different AP ccessions in su-min plots with three replictes. Seventy-dy-plnts (efore flowering stge) were exposed to different slinity levels on Hoglnd medium. The results indicted tht slinity levels hd significnt effect on the mesured trits. Slinity cused reduction in dry mteril nd slt tolernce index (STI). The sodium (N + ), iron (Fe), zinc (Zn) nd copper (Cu) contents significntly incresed, while nitrogen (N), phosphorus (P), potssium (K + ), clcium (C 2+ ), mgnesium (Mg 2+ ) nd mngnese (Mn) content decresed in high slinity levels. Furthermore, under slinity stress, tolernt ccessions could produce higher K +, P, N, Mg 2+ nd C 2+ nd lower N +, Fe, Zn nd Cu thn sensitive ccessions. Interestingly, N + concentrtion ws highly correlted to STI (r =.79), while the lowest correltion ws oserved etween P nd N + concentrtion (r =.15). Sltstress cused chnges in mcro nd micro nutrients uptke, which my led to decline in photosynthesis cpcity nd respirtion. This type of iotic stress cn lso distur metolism of severl cellulr components such s protein synthesis. Keywords: Androgrphis pnicult, slinity stress, slt tolernce, mcro nd micronutrients. Arevitions: CHLO, chlorophyll; NL, numer of leve; RL, root length; SL, shoot length; STI, slt tolernce index; TDW, totl dry weight. Introduction Soil slinity is mjor environmentl fctor cuses reduction in plnt growth nd productivity in rid nd semirid res of the world. Approximtely, 2% of the world s cultivted re nd out hlf of the world s irrigted lnds re reported to e seriously ffected y slinity nd wter logging (Munns nd Tester, 28; FAO, 28). In generl, high slt concentrtion in soils cuses reduction in osmotic potentil, which led to distur wter vilility to root cells nd mke difficulties for plnt to otin oth wter s well s nutrients. Consequently, rpid reduction hppens in growth rte, productivity nd mny metolic chnges due to hormonl signl generted y the roots (Munns, 22; Ashrf nd Hrris, 24). Reserchers hve indicted tht plnt species hve different ilities to tolerte slinity stress, which depends on severl intercting vriles such s the plnt growth stge, slt concentrtion nd the durtion of stress over time (Munns, 22; Vllejo et l., 21; Zhu, 22). The deleterious effects of slinity on plnt growth is ttriuted to the decrese in osmotic potentil of the growing medium, specific ion toxicity nd nutrient ion deficiency y disrupting potssium (K + ) nutrition, which cn e result of inorgnic ion (N +, Cl, nd K + ) nd comptile orgnic solute ccumultions (solule crohydrtes, mino cids, proline, etines, etc.) (Luo et l., 25; Hsegw et l., 2). The osmotic djustment in oth roots nd leves contriute to the mintennce of wter uptke nd cell turgor, llowing physiologicl processes, such s stomt opening, photosynthesis, nd cell expnsion (Serrj nd Sinclir, 22). It is reported tht the root growth nd totl fresh weight of root were more dversely ffected y slinity compred to shoot growth (Prid nd Ds, 25). Eventully, responses of plnt roots nd shoots to soil slinity should e understood oth under slinity conditions. Besides, there is the possiility tht mny nutrient interctions in sltstressed plnts my hve importnt consequences to growth. Plnts require mcro nd micro elements to grow. In soil, micronutrients re not lwys present in the solution nd their 271
vilility is limited due to severl fctors, which minly limit their soluility. The min fctors determining the level of solule Mcro- nd micronutrients in soil solution re: Oxidtion-reduction rections nd the ph. Some of the interctions interfere nd dmge the vilility of micronutrients. Some of the interctions re soil interctions, nd some others hppen in the plnt tissue due to excessive concentrtions. Internl concentrtions of mjor nutrients nd their uptke hve een frequently studied (Ms nd Grieve, 1987; Crmer et l., 1987), ut the reltionship etween mcro nd micronutrient concentrtion under slinity condition is rther complex nd remins poorly understood (Tozlu et l., 2). Androgrphis pnicult elongs to fmily Acnthce is n importnt medicinl plnt (Vldini et l., 212). The plnt exhiited spcious cpcity of phrmceuticl properties such s nti-hiv (Bsk et l., 26), nti-h1n1 (Ko et l., 26), nticncer (Chun et l., 21) nd nti-heptitis (Dumrongsk et l., 29). It grows undntly in southestern Asi in moist nd sunny situtions (Lttoo et l., 28). It is n erect nnul her with drkgreen qudrngulr stem; lnceolte nd pinnte leves; smll nd white flowers; liner-olong cpsules; nd tiny yellowish-rown seeds (Jing et l., 29). A huge lck of informtion relting to the slt-tolernce threshold nd modlity of minerl nutrition of this her t sline re is oviously sensile. Moreover, the slinity effects on growth nd yield, hs direct economic implictions. On the other hnd, A. pnicult is ctegorized in the group of sensitive plnts to slinity. The symptoms of such sensitivity pper y scorched lef tips nd mrgins, lef curling, nd in extreme cses reduced growth, scission of leves, nd deth of the plnt. However, reports on mcro nd micronutrients uptke rtes re ville within plnts, ut the results hve not lwys een consistently relile. The reltionship etween slt tolernce nd Mcro-micro elements tendency of the plnt is not investigted yet. Determintion of slinity effects on mcro-micro elements nd the interreltionships mong trits will help to mke etter comprehension of growing inquiries nd the mechnisms of slt tolernce of the plnt under slinity. The present explortion is sort of first ttempt to understnd the dptive fetures of the quntittive effects of slinity levels on Mcro-micronutrients flow in A. pnicult. Results Assessment of morphologicl trits efore pplying slinity stress The results of our study showed tht there were no significnt differences mong 12 ccessions for SL, NL, TDW nd chlorophyll content. The low coefficient of vrition (CV = 4.1-1.1 %) efore pplying slinity stress complied with the ccurcy of the experiment nd homogeneous feedck of the studied ccessions (Tle 1). Evlution of different ccessions of A. Pnicult t control conditions The ANOVA nlysis of the investigted chrcteristics under control condition reveled significnt difference mong the studied ccessions in terms of ll Mcro nd micronutrients uptke, except for the Fe nd Mn nutrients. It lso showed tht there is no significnt difference in totl dry weight of the 12 ccessions (Tle 2). This outcome, in fct evidenced tht the selected ccessions for this experiment were significntly different from ech other in the uptke cpcity of microelements even under norml condition. Effects of different slinity levels on yield nd STI The slinity levels ffected the yield of A. pnicult. Vrition due to slinity levels (S), ccessions (A) nd their interction were highly significnt (p<.1). The interction of S A ws not significnt in terms of totl dry weight (TDW), slt tolernce index (STI) (Tle 3). Totl dry weight results showed tht growth ws negtively correlted to the sustrte concentrtion of NCl (p<.1). Plnts grown t the low levels of NCl (control) reched reltively to the higher totl dry weights nd did not exhiit the toxicity symptoms. However, the totl dry weight ws significntly reduced t higher levels of slinity (16 dsm -1 ), in which the symptoms of slt toxicity ws reveled s growth depression. The leves commenced to fll on the dy 2 th of pplying 16 dsm -1 of slinity. The concentrtion of 16 dsm -1 NCl significntly reduced the dry weight y 29.49%, compred to control. Among the ccessions, the highest STI (68.7%) ws oserved in ccession 11264, wheres the lowest (49.1%) with the sme condition elonged to ccession 11329 (Fig. 1). At high slinity level, the slt tolernce index of tolernt ccession (11264) ws found to e higher (43.19%) thn the sensitive one (11329) with only 18.23%. The gretest decrese in STI ws oserved t 16 dsm -1 of slt stress. Effects of different slinity levels on mcronutrients The nlysis of vrince reveled significnt difference etween slinity levels, in terms of mcronutrients ccumultion in the plnts. Vrition due to the slinity levels (S), ccessions (A) nd their interction were highly significnt (p<.1). The interction of S A ws not significnt in terms of clcium content (C 2+ ) (Tle 3). The slinity levels significntly decresed ll mcronutrient contents (Fig. 2 nd Fig. 3). The slinity levels significntly cused chnges in performnces of ll ccessions. All ccessions showed decresed in K + content under slt tretment. The potssium content of tolernt ccession ws found to e higher thn sensitive one in the slt medium. Moreover, the clcium (C 2+ ) ccumultion of different ccessions decresed under slt tretment. The sline tretment lso decresed the other mcronutrients (P, N nd Mg 2+ ) in ll ccessions compred with control. The decreses in tolernt ccessions were lower thn sensitive ccessions. The potssium contents of ccession 11264 nd 11179 were found to e higher thn ccession 11329 nd 1136 in the slted medium. The high slinity level of 16 dsm -1 significntly reduced the K + content y 81.8 %, compred to control. The phosphor, nitrogen, clcium nd mgnesium contents of tolernt ccession(11264) were found to e higher thn the sensitive one(11329) under high level of slinity (Tle 4). Effects of different slinity levels on micronutrients The slinity levels significntly ffected ll ccessions in terms of N + nd Zn content, while there were no significnt differences mong ccessions sed on Cu, Fe nd Mn (Tle 5). The interction of S A ws not significnt in terms of Fe concentrtion. The slinity levels significntly incresed micronutrients except Mn (Fig. 2 nd Fig. 4). Among the 272
Tle 1. Anlysis of vrince on mesured chrcteristics of 12 ccessions of A. pnicult efore slinity stress t mture stge. Source df Men Squre NL SL TDW CHLO R 2 1.5 * 12.35 ns.75 *.51 ns Accession 11.48 ns 3.16 ns.22 ns 2.7 ns Error 22.54 4.56.11 4.51 CV% 4.8 8.7 1.1 4.1 * nd ns, refer to 5% nd not significnt, respectively. NL: numer of leve, SL: shoot length, RL: root length, TDW: totl dry weight nd CHLO: chlorophyll. Fig 1. The effects of slinity levels on slt tolernce index in 12 cessions of A. pnicult. Error rs show pproximtely 99% confidence intervls nd different letters indicte significnt difference mong ccessions using Duncn s multiple comprison test t P <.1. ccessions, the highest N + ccumultion (7.3%) ws oserved in ccession 11329, wheres the lowest N + ccumultion (4.44%) elonged to ccession 11264 oth t sme condition (Fig. 2). In ll ccessions N +, Cu, Zn nd Fe content incresed under the effect of slt tretment, while Mn content decresed t the sme condition (Tle 4). In the high slinity level, some ccessions such s 11264 nd 11149 showed significntly less N + thn other ccessions (11329 nd 1136). Correltion mong the trits under slinity stress condition The correltion mong most of the mesured trits ws significnt t p<.1 (Tle 6 nd Fig. 5). Interestingly, N + concentrtion ws highly correlted to STI, while others not. The correltion etween STI nd N + s well s Zn nd Fe concentrtions were negtively significnt, while positive correltion ws determined etween STI nd K +, C 2+, Mg 2+, N nd Mn concentrtions under slt stress. There ws no significnt correltion etween STI nd P nd Cu content. Cluster nlysis The cluster nlysis of the 12 ccessions of A. pnicult using UPGMA-Wrd method in control conditions produced two clusters for STI nd ll mcro nd micronutrients (Fig. 6), while under slinity conditions four clusters creted for STI nd ll nutrients (Fig. 7). Cluster 1 (red color) contined three ccessions including tolernt ccession, cluster 2 (green color) contined three ccessions, cluster 3 (lue color) contined three ccessions including sensitive ccession nd cluster 4 included three ccessions. The distnce etween leder nd joiner ccession in cluster nlysis ws 4.45 sed on ll mesured trits. According to cluster nlysis, the studied ccessions rected in different wys under control nd slt conditions, where numer of clusters in control ws two nd then incresed up to four clusters under slt stress. In fct, the chnge in numer of clusters hppened due to the slt tretment. Discussion After 3 dys of tretment, the results indicted tht pplied NCl inhiited the plnt growth, nd led to decrese in dry mteril in the slted medium compred to control. This my e relted to the effect of slt stress, which cuses the limittion of wter sorption nd iochemicl processes. A liner decline in dry mteril correlted with incresing slinity levels ws recorded in our experiment. This result ws in greement with mny other reports on the other plnts, in which the dry weight under slt-stress ws less in the control medium (Pek et l., 1988; Liu nd Vn Stden, 2; Munns, 22; Prid nd Ds, 25). Sensitive ccessions under slinity condition produced less dry weight thn tolernt ccessions. For instnt, ccessions 11264 (tolernt) produced 31.3% less dry weight in high slinity level, while the sensitive ccessions such s 11329 produced 5.9 % less dry weight compred to control. Severl 273
N, K nd N content (percent) C, Mg nd P content (percent) Tle 2. Anlysis of vrince on nutrient chrcteristics of 12 ccessions of A. pnicult in control condition. Source df Men squre TDW K + P N C 2+ Mg 2+ N + Cu Zn Fe Mn Accession 11 22.4 ns.21 **.8 **.58 **.5 *.5 **.2 ** 24.8 ** 493.3 * 474.7 ns 588.6 ns Error 22 1.7.5.1.13.2.1.4 5.9 156.5 2632.1 469.6 CV% 1.1 9.2 12.7 7.2 24.2 11.3 21.8 25.8 18. 39.2 32.5 Tle 3. Anlysis of vrince on yield, STI nd mcronutrients in 12 ccessions of A. pnicult under slinity stress. Men squre Source df TDW STI K + P N C 2+ Mg 2+ Slinity (S) 4 3179.4** 3474.4** 1.59**.25** 6.85**.24**.42** Accession (A) 11 5.9** 473.9**.12**.9**.52**.8*.4** S A 44 1.9ns 96.9ns.12**.5**.44**.5ns.3** Error 19 1.6 13.2.5.2.21.3.2 **, * nd ns, refer to 1%, 5% nd not significnt, respectively. TDW: totl dry weight nd STI: slt tolernce index 6 5 4 3 2 1 N K N e d c c c Control 4 8 12 16 Slinity levels (dsm -1 ).7 Mg C P.6.5.4.3.2.1. c c c Control 4 8 12 16 Slinity levels (dsm -1 ) Fig 2. The effects of slinity levels on trend of N + nd K + ccumultion of A. pnicult. Verticl rs represent S.E. for three smples nd different letters indicte significnt difference mong the slinity levels using Duncn s multiple comprison test t P <.1. Fig 3. The effects of slinity levels on mcro nutrients ccumultion in A. pnicult. Verticl rs represent S.E. for three smples nd different letters indicte significnt difference mong the slinity levels using Duncn s multiple comprison test t P <.1. reserchers, hve reported similr differences etween sensitive nd tolernt genotypes in different plnt species (Munns, 22). The K + is required for cells to mintin the osmotic lnce, opening nd closing of stomt nd n essentil cofctor for mny enzymes (Yeo, 1998). High stomt K + requirement is reported for photosynthesis (Chow et l., 199). The result of the present study showed tht NCl tretment cused n increse in N + concentrtion, nd decrese in K + nd C 2+ concentrtion in ll ccessions. Seemingly, this result is similr to findings of the other reserchers (Colmer et l., 25; Mtuis nd Amtmnn, 1999; Munns et l., 22). However, the increse in N + concentrtion in sensitive ccession like 11329 ws higher thn tolernt ccession such s 11264, ut the decrese in K + nd C 2+ concentrtion in sensitive ws higher thn the tolernt ccession. Clcium is importnt during slt stress, e.g. in cell wll nd memrnes, nd regultes growth nd development of plnt (Hepler, 25). As result, C 2+ fertilizers my reduce N + toxicity to the plnts. Besides, the role of Mg 2+ in chlorophyll structure nd s n enzyme co-fctor is considerle. Another importnce of Mg 2+ in plnts is in the export of photosynthetic, which leds to improve chlorophyll reduction in Mg 2+ deficient source leves (Mrschner nd Ckmk, 1989). In generl, slinity reduces N content in plnts (Feigin, 1985). The interction etween slinity nd P is very complex nd there is no cler mechnism to expln the chnges of P uptke in response to slinity stress in different species (Grttn nd Grieve, 1992). However, it is known tht P concentrtion is relted to the photosynthesis rte (Overlch et l., 1993) nd; therefore, decrese of P in leves will reduce the plnt growth. In sline soils, the soluility of micronutrients (Fe, Zn, Mn, nd Cu) is minly low, nd plnts grown in these soils often develop deficiencies in these nutrients (Pge et l., 199). Differences cn e ttriuted to plnt type, plnt tissue, slinity level nd environmentl conditions. Thus, slinity stress either promotes or inhiits the uptke of some micro nutrients depends on plnt species. The Fe, Zn nd Cu content increse in response to rising NCl levels, which is consistent with other reserchers tht reported high uptke of these micronutrients (Moreno et l., 2; Alm, 1999). In present study, we found similr tendency of incresed concentrtions of ll micronutrients. Nevertheless, the Mn ws n exceptionl cse in this pproch. Mngnese is importnt in ctivity of enzymes involved in respirtion nd synthesis. 274
Fe, Zn nd Mn content (ppm) Tle 4. The effects of slinity levels on mcro nd micronutrients in 12 ccessions of A. pnicult. Accession K + % P% N% C 2+ % Mg 2+ % N + % Zn (ppm) 11179 2.3±.8 c.17±.2 4.7±.17 cd.61±.4 c.24±.1 cd 2.±.4 93.1±5. c 11216 2.2±.5.18±.2 4.4±.15 c.57±.7 c.23±.1 cd 2.4±.5 cd 78.1±2.6 11228 2.2±.6.21±.2 cde 4.4±.12 c.39±.5.19±.1 2.5±.5 cd 72.9±3. 11249 2.2±.6 c.2±.2 cd 4.6±.19 cd.45±.5.2±.1 2.2±.4 76.6±4. 11264 2.1±.6.19±.2 c 4.3±.16.53±.5 c.22±.1 cd 2.3±.4 c 73.2±3.1 11265 2.2±.8.2±.2 c 4.7±.18 cd.43±.6.2±.2 c 2.1±.4 8.4±4.5 11266 2.2±.8.2±.2 cd 4.6±.15 cd.55±.5 c.23±.1 cd 2.4±.6 c 78.5±4.3 1136 2.2±.11.25±.2 f 4.7±.24 cd.44±.5.21±.1 c 2.4±.6 c 66.6±3.6 11314 2.4±.13 c.23±.1 cdef 4.8±.21 cd.5±.4 c.22±.2 c 2.5±.6 cd 75.7±5.6 11329 2.1±.9.24±.2 ef 4.6±.16 cd.54±.5 c.22±.1 c 2.9±.7 d 77.2±4.6 11339 2.1±.9.2±.2 cd 4.2±.18.63±.6 c.25±.1 2.8±.7 cd 78.6±6.2 11348 2.2±.7.23±.2 def 4.8±.13 d.48±.7 c.21±.1 c 2.8±.7 cd 83.3±5.6 c Different letters indicte significnt difference etween the vlues of pirs of tretment within columns (men vlues ± stndrd error) t P <.1 ccording Duncn s multiple comprisons test. Tle 5. Anlysis of vrince on micronutrients in A. pnicult under slinity stress. Source df N + Men squre Cu Zn Fe Mn Slinity (S) 4 166.69** 23.46** 1285.33** 42579.3* 3217.18** Accession (A) 11 1.21** 47.6ns 614.52** 4263.75ns 159.87ns S A 44.73** 68.28** 352.13** 442.89ns 527.14** Error 19.38 26.53 195.66 4434.5 266.7 **, * nd ns, refer to 1%, 5% nd not significnt, respectively. 2 18 16 14 12 1 8 6 4 2 Zn Mn Fe c c c c c c Control 4 8 12 16 Slinity levels (dsm -1 ) Fig 4. The effects of slinity levels on micronutrients ccumultion in A. pnicult. Verticl rs represent S.E. for three smples nd different letters indicte significnt difference mong the slinity levels using Duncn s multiple comprison test t P <.1. Fig 5. Correltions mong 12 ccessions for STI, mcro nd micronutrients under slinity condition. The strength nd direction of the correltions mong the different trits re indicted y the color (red indictes positive correltions while green indictes negtive correltions, nd the shding represents the strength of the correltion). It is difficult to suggest mechnism to explin the slinity impcts on micronutrients due to the reltively smll differences etween control nd slt-treted plnts nd the non-liner reltionships etween some of the micronutrient contents in plnts. Mterils nd methods Genetic mteril nd germintion conditions Twelve ccessions of A. pnicult seeds provided from the Agro Gene Bnk, University Putr Mlysi, used for this study during Septemer 21 - April 211 (Tle 7). The seeds were surfce sterilized with 1% sodium hypochlorite (NOCl) solution for 1 minutes (Tlei et l., 211) nd thoroughly rinsed with distil wter. Then, seeds were soked in 15 cm dimeter seprte Petri dishes contining filter ppers No.2 moistened with sterile wter. The Petri dishes were seled with prfilm to prevent ny wter loss nd then incuted under controlled growth chmer (light/drk regime of 14/1h t 28-3 o C, reltive humidity 6-75%). The seedlings t two leves stge were trnsferred into the jiffy medi. 275
Tle 6. Phenotypic correltion coefficients (r) mong mesured trits in 12 ccessions of A. pnicult under slinity conditions. STI N + % K + % C 2+ % Mg 2+ % P% N% Cu (ppm) Zn (ppm) Fe (ppm) Mn (ppm) STI 1 N + -.785** 1 K +.478** -.53** 1 C 2+.26** -.97.22** 1 Mg 2+.466** -.291**.43**.752** 1 P.17 -.15*.174*.2.111 1 N.421** -.435**.53** -.16.14.254** 1 Cu -.143.131 -.3.74.76 -.72 -.81 1 Zn -.326**.233** -.31 -.33 -.28.1 -.12.228** 1 Fe -.293**.35** -.227**.78 -.4.177* -.117.78.347** 1 Mn.296** -.277**.184* -.154*.88.277**.366**.37.137.79 1 The significnt correltion is indicted y ** (P<.1) nd * (P<.5). Tle 7. List of 12 ccessions of A. pnicult No. Accession numer Stte Vernculr nme 1 11179 Selngor Tutup Bumi 2 11216 Negeri Semiln Hempedu Bumi 3 11228 Negeri Semiln Hempedu Bumi 4 11249 Negeri Semiln Hempedu Bumi 5 11264 Perk Akr Cerit 6 11265 Perk Akr Cerit 7 11266 Perk Akr Cerit 8 1136 Johor Hempedu Bumi 9 11314 Terenggnu Lidh Ulr 1 11329 Kelntn Lidh Ulr 11 11339 Kelntn Lidh Ulr 12 11348 Terenggnu Lidh Ulr Fig 6. Dendrogrm generted y UPGMA clustering method of 12 ccessions of A. pnicult sed on STI, mcro nd micronutrients t control condition. Fig 7. Dendrogrm generted y UPGMA clustering method of 12 ccessions of A. pnicult sed on STI, mcro nd micronutrients under slinity condition. Tolernt nd sensitive ccessions were plced in the red nd lue clusters, respectively. Experimentl design We crried out split plot experiment sed on Rndomized Complete Block Design (RCBD) with two fctors nd three replictes. The fctors were five different concentrtion of sline wter [control (), 4, 8, 12 nd 16 dsm -1 ] in min plots nd 12 different ccessions in su min plots. The 3-dy-seedlings were trnsferred from jiffy medi into the pot with snd medium. After 3 dys from culturing (lmost in 7 dys old), the plnts were plced in different slinity levels (, 4, 8, 12 nd 16 dsm -1 ) on Hoglnd medium. Ech plnt ws irrigted once dy with five levels of sline wter. After every three slinity pplictions, plnts were gin irrigted with norml Hoglnd nutrient solution. After 3 dys slinity exposure, ll plnts were hrvested nd dt on totl dry weight (TDW) (fter drying t 68 o C for 48 hours), slt tolernce index (STI), N +, K +, C 2+, Mg 2+, N, P, Cu, Zn, Fe nd Mn content were mesured. Determintion of mcro nd micronutrients content Nitrogen ws determined y the digestion method descried y (AOAC, 199) using n utomted ion nlyzer (Lcht instrument, Wisconsin, USA) nd others mcro nd micronutrients content were determined y using n Anlyst 51, Perkin Elmer instrument, USA descried y Miller (1998). 276
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