Fatemeh Rezaie Kinji 1 *, Parvaneh Rahdari 2. Research Article. Greener Journal of Agricultural Sciences

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1 Greener Journl of Agriculturl Sciences ISSN: ; ICV: 6.15 Vol. 2(7), pp , Novemer, 2012 Copyright 2017, the copyright of this rticle is retined y the uthor(s) Reserch Article Shortge on (Mgnesium nd Clcium) Dry Weight, Fresh Weight, Root nd Shoot Length, Lef Reltive Wter Content (RWC), Chlorophyll Content nd Mlondildehyde Activity in Fenugreek (Trigonell Foenum Grceum) Ftemeh Rezie Kinji 1 *, Prvneh Rhdri 2 1 High Student of Plnt Physiology of Islmic Azd University of Tonekon Brnch 2 Assistnt Professor nd Fculty Memer of Islmic Azd University of Tonekon Brnch ARTICLE INFO ABSTRACT Article No.: DOI: /GJAS Sumitted: 08/10/2012 Accepted: 20/10/2012 Pulished: 30/11/2012 *Corresponding Author Ftemeh Rezie Kinji E-mil: ftemerezei16@yhoo.com Keywords: Fenugreek (Trigonell Foenum Grceum), Minerl Element, RWC, Growth Fctors, Mlondildehyde Fenugreek (Trigonell Foenum Grceum) is one of the medicinl plnts tht hve een used s folk medicine in Irn nd mny countries. In this study, effects of mcronutrient element shortge (Nitrogen, Phosphorous, Potssium, Mgnesium nd Clcium) on dry nd fresh weight, root nd shoot length, Lef Reltive Wter nd chlorophyll content in Fenugreek plnt ws investigted. In reltion to Mgnesium shortge which cused, increse of root nd shoot dry weight, root nd shoot fresh weight, root nd shoot length nd lef reltive wter content, it ws oserved tht this increment ws meningful in 0/05 of sttisticl level. In Clcium shortge, root nd shoot length, root nd shoot dry weight nd shoot fresh weight decresed, which in sttisticl level 0/05 showed meningful difference thn lnk level, while in reltion to lef reltive wter content, there ws no meningful difference to the control level. In Potssium shortge, shoot length, root nd shoot fresh weight nd lef reltive wter content meningful incresed, while root nd shoot dry weight, oserved decrement in sttisticl level of 0/05 which ws meningful thn lnk level. Investigting the effect of Phosphorous shortge, shoot length meningfully incresed thn in lnk level, while in root dry weight, shoot fresh weight nd lef reltive wter content, meningful decrement ws oserved thn in the lnk level (P<0.05). In Nitrogen shortge, root length nd fresh weight nd lef reltive wter content meningfully incresed thn lnk, while shoot fresh weight, root nd shoot dry weight were meningfully decresed in sttisticl levels of 0/05 thn lnk. Mcronutrient elements shortge cused decrement of chlorophyll in Fenugreek leves with Nitrogen, Potssium nd Phosphorous recording shortge thn the lnk level. In plnts with Clcium nd mgnesium shortge, increse of chlorophyll ws oserved. Mcronutrient elements shortge cused meningful decrese in chlorophyll nd totl chlorophyll concentrtion in Fenugreek leves (P<0.05). Also, mcronutrient elements shortge cused decrese in Mlondildehyde ctivity in root nd shoot of Fenugreek plnt.

2 Ftemeh nd Prvneh / Greener Journl of Agriculturl Sciences 295 INTRODUCTION Fenugreek(Trigonellfoenum L.) is n ngiosperm plnt. This plnt hs dicotyledonous petls from the order of roses, pes groups, sugroups of groups nd genus (Trigonel L.) from group trifolite. The nme of this plnt is comined from the Greek word trigon nd Foenumgrecum. Fenugreek is herceous nd n nnul plnt tht hs its height t 50cm. Fenugreek is one of the medicinl plnts tht hve een used in Irn nd severl ntions trditionl medicines nd it hs mny helth enefits. Fenugreek hs wide rnge of therpeutic effects such s nlgesic, nti-therosclerotic, ntiinflmmtory, crmintive, ntispsmodic, nti-cncer, chelting lood sugr, incresing sexul desire, stringent, crdic tonic, ldder, lxtive, expectornt diuretic, reducing cholesterol, reducing lood lipids, reducing lood pressure, uterine tonic, nti-worm. Growth nd yield of griculturl plnts is function of environmentl fctors nd their interctions. Lowit (1980)suggests tht stress is result of norml physiologicl processes otined from the effects of comintion of environmentl fctors. Actully, the mount nd severity of these dverse fctors cn cuse prolems in orgnisms nd lso direct nd indirect dmges in plnt or its components. These limiting fctors re clled environmentl stress nd they re divided into two types: iologicl nd non-iologicl (Meyodi,1381).Mture stge hs significnt effects on plnt minerls nd one of the most importnt influences is mture plnts with high phosphorus reduction. Nitrogen, phosphorus nd potssium in young plnts nd young tissue plnts re high. On the other hnd, clcium, mngnese nd iron re greter in older plnts nd more mture prts of the plnt. Levels of minerl will chnge with incresing plnt ge. Clcium increses with incresing plnt ge while P reduces with incresing plnt ge. In different regions of world, concentrtion of minerls in plnts, depend on the interction of severl fctors such s soil, plnt species, nd stge of growth, climte, nd production nd interction elements in the sorption time. Plnts reduce the hrmful effect of stress with increse in metolism nd sets osmotic potentil y the ccumultion of orgnic nd inorgnic mterils in the cells.this method works y the sorption of inorgnic ions y plnt in the externl environment such s incresing the concentrtion of potssium in the shoots nd/or synthesis of comptile solutes y high concentrtions of mterils such s osmolyte In response to stress, chnges creted in the ionic current. Sodium including elements tht will ccumulte when pplied to environmentl stress in the plnt nd it cn led to disrupted memrne nd inhiition of ion trnsport of nutrients from roots to shoots. Mgnesium is minerl component of the chlorophyll molecule. Mgnesium contriutes in the construction of the plnt oil nd it sets phosphorus uptke in the plnt nd lso it is effective in hydrocron nd sugr production. One of the symptoms of mgnesium deficiency in plnts is the yellowing etween the veins. First, the symptoms of deficiency re oserved in old leves nd leves fll y cute shortge. Clcium plys n importnt role in the stility of the cell wll, cell development nd internl processes, memrne stility, lnce of ctions nd nions, ctivtor of some enzymes nd osmotic pressure. Clcium deficiency cuses nlysis of cell wll nd dried leves. This element is necessry for synthesis nd trnsport of nutrients in plnts nd elimintes some dverse effects of nutrient imlnces of plnts in soils. Also this element is effective in the regultion of wter uptke. Phosphorus ecomes unusle ecuse of their specific rections such s soil dsorption nd sediment formtion for plnt. In clcreous soils nd clcreous plster (soils of Irn), one of the prolems of plnt nutrition in the soils is the lck of plnts with ville phosphorus ecuse conversion of solule phosphorus to less solule compounds such s clcium phosphtes. Therefore the entrnce of P in different rections with composition of soil nd low phosphorus vilility in soil, mounts of this element is necessry for pproprite concentrtions of growth in soils (Govere, 2004). MATERIALS AND METHODS Fenugreek seeds were prepred from the reserch institute of forest nd rngelnd nd study ws performed in the plnt iology lortory in Islmic Azd University of Tonekon in spring 1390.Produced seeds hd the sme size. For disinfection of seeds, we used 5% sodium hypochlorite (Distilled Wter 5%) in 10 minutes nd then seeds were wshed with distilled wter (Flhti, 2007). For germintion, seeds were cultured in medium Petri dishes in germintor with temperture of 25 C. After four dys of growth, seeds were plnted in pots with 30 cm dimeter nd length with snd plnted. Then, pots were plced in growth room, with temperture of 1 ± 23 (1 ± 26 c during the drk nd light periods) nd light intensity 25 kilo lux nd 16-h photoperiod. In the first week fter plnting in pots, irrigtion ws performed with 40 ml of distilled wter. After the first week order to supply of minerl, irrigtion ws performed y 40 ml of distilled wter with Hoglnd solution. After 13 dys ccording to the deficiency, 40 ml prepred solution ws given to ech pot. Numer of pots for 5 tretments with 4 replictes nd control group were included. Tretment of Plnt After 7 dys, Hoglnd solution with PH of 5/7 ws given to ll pots shown in tle 1. Also, the mteril requirements expressed in tle 1 is to pply for ny deficiency (Norikne, 2003). Order to evlute dry weight,

3 296 Ftemeh nd Prvneh / Greener Journl of Agriculturl Sciences seedlings were plced in n oven with temperture of 50 C nd fter 24 hours the dry weight ws mesured. Sttisticl Anlysis Studies were conducted in completely rndomized design with four replictes nd dt nlysis with nlysis of vrince nd Duncn's men comprison test reviewed y SPSS nd chrt drwn with Excel. mm /67 0/05 5 0/05 0/5 16/5 0/1 0/05 0/05 Tle 1: Hoglnd solution for supplying chemicls nd imposed deficiencies Control N C Mg K KNO3 K2SO4 KNO3 KNO3 NNO3 C(NO3)2 CCl2 NNO3 C(NO3)2 C(NO3)2 MgSO4.7H2O MgSO4.7H2O MgSO4.7H2O N2 SO4 MgSO4.7H2O NH2PO4.2H2O NH2PO4.2H2O NH2PO4.2H2O NH2PO4.2H2O NH2PO4.2H2O Fe-EDTA Fe-EDTA Fe-EDTA Fe-EDTA Fe-EDTA Mn SO4.H2O Mn SO4.H2O Mn SO4.H2O Mn SO4.H2O Mn SO4.H2O Cu SO4.5 H2O Cu SO4.5 H2O Cu SO4.5 H2O Cu SO4.5 H2O Cu SO4.5 H2O Zn SO4.7H2O Zn SO4.7H2O Zn SO4.7H2O Zn SO4.7H2O Zn SO4.7H2O H3BO3 H3BO3 H3BO3 H3BO3 H3BO3 N MoO4.2H2O N MoO4.2H2O N MoO4.2H2O N MoO4.2H2O N MoO4.2H2O CoSO4.7H2O CoSO4.7H2O CoSO4.7H2O CoSO4.7H2O CoSO4.7H2O Ni SO4.7H2O Ni SO4.7H2O Ni SO4.7H2O Ni SO4.7H2O Ni SO4.7H2O P KNO3 C(NO3)2 MgSO4.7H2O NCL Fe-EDTA Mn SO4.H2O Cu SO4.5 H2O Zn SO4.7H2O H3BO3 N MoO4.2H2O CoSO4.7H2O Ni SO4.7H2O After 42 dys of plnting the seeds in pots nd 5 periods of tretment, plnts were removed from pots nd then the root length, length of shoot, root fresh weight, shoot fresh weight, reltive wter content, mesurement of chlorophyll nd mesurement of mlondildehyde ws clculted. Reltive Wter of Shoot in Treted Plnts nd Control First shoots were plced in two distilled wter for 5 hours in Petri dish. After torgor, weight of torgor ws mesured with digitl scles nd the numers were plced in following formul. It ws clculted sed on Smrt nd Binghm DW =Dry weight FW =Fresh weight TW =Torgor weight Mesurement of MDA Totl size MDA ws mesured ccording to the method of Ohkw According of this method, 01.0 g from fresh plnt tissue (Root / Shoots) with Trichlorocetic cid 5% (TCA) ws pulverized inside mortr equl mnner. Solule moved the micro tue nd seprte opertion ws performed in rpm12000 centrifuge for 15 minutes. Thiorituric 5% ws dissolved in TCA 20% nd moved to test tues for 25 minutes t temperture of 100 C. Then, these tues were centrifuged t rpm for 5 minutes. Otined solution uptke red t 532 nm nd for review, dt ws red during the 600 nm wvelength. Extinction coefficient for MDA ws 155 mm cm. = A =Asorption = Extinction Coefficient B =Mesured Cell Width C=the Amount of Mteril Mesurement of Chlorophyll Content The method of Arnon (1949) ws used to mesure chlorophyll. 1.0mg of fresh plnt hs een homogenized in the Chinese mortr y Stn. In order to prevent the withdrwl of mgnesium in the chlorophyll uilding, clcium cronte 5/0mM ws dded. The otined solution ws filtered with Wtmn pper No. 2. Extrct dilution ws performed y 80% cetone. To clculte the mount of chlorophyll, sornce ws determined in the wvelengths of 645 nd 663nm using 80% cetone control. Lef chlorophyll content per mg per g wet tissue ws clculted using the following formul:

4 Ftemeh nd Prvneh / Greener Journl of Agriculturl Sciences 297 C=(0/0127.A(663)-0/00269.A(645)mgr/mLit C=(0/0229.A(645)-0/00486.A(663) mgr/mlit CT=(0/0202.A(645)-0/0082.A(663) mgr/mlit RESULTS Severl studies hve een performed on plnts nd the effects of nutrient stress on the growth fctors t seedling stge tht it shows drop in growth of stress. In this study, results showed reduce stress effects on growth. Significnt difference ws oserved mong the tretments during rdicle nd the highest growth rte ws oserved in plnts deficient in nitrogen nd mgnesium in comprison with control (Figure 1). Chnges in length of shoot, the effect of nutrient deficiencies showed significnt increse (p <0.05) in plnts with mgnesium deficiency thn in control. Wheres significnt decrese ws oserved in plnts with clcium deficiency thn in the control; ut the difference ws not significnt in plnts with phosphorus nd potssium deficiency thn the control (Figure 2). The root dry weight ws significntly different (p <0.05) mong the different tretments nd ccording to the oservtion on plnts with mgnesium deficiency thn in the control showed significnt increse, while the difference ws not significnt in clcium deficiency thn in the control (Figure 3).Also, root fresh weight showed significnt increse thn control in the effects of mgnesium deficiency(p <0.05) wheres there ws no significnt difference in clcium deficiency thn control (Figure 5).Shoot fresh weight showed the highest increse thn control in mgnesium deficiency plnts nd weight ws equl in plnts with nitrogen nd clcium deficiency(figure 6). According to numers clculted in the ove nmed formul, it ws found tht reltive wter content shoots hd significnt increse in plnts potssium deficiency thn control wheres it ws equl in plnts with clcium nd mgnesium deficiency nd hd significnt decrese in plnts with phosphorus deficiency thn control (Figure 7). Figure 1: Effect of high-nutrient deficiency on root of Fenugreek Height (cm) Control K Mg N P C Tretments Figure 2: Effect of high-nutrient deficiency on shoot length Fenugreek

5 298 Ftemeh nd Prvneh / Greener Journl of Agriculturl Sciences Figure 3: Effect of high-nutrient deficiency on dry weight of Fenugreek s root Figure 4: Effect of high-nutrient deficiency on dry weight of Fenugreek s shoot Figure 5: Effect of high-nutrient deficiency on fresh weight of Fenugreek s root

6 Ftemeh nd Prvneh / Greener Journl of Agriculturl Sciences 299 Figure 6: Effect of high-nutrient deficiency on fresh weight of Fenugreek s shoot Figure 7: Effect of high-nutrient deficiency on reltive wter of Fenugreek s lef Totl Chlorophyll (mg/m lit) Figure 8: Effect of high-nutrient deficiency on Chlorophyll of Fenugreek Control K Mg N P C Tretments

7 300 Ftemeh nd Prvneh / Greener Journl of Agriculturl Sciences MDA (mm -1 cm -1 ) Control K Mg N P C Tretments Figure 9: Effect of high-nutrient deficiency on Mlondildehyde of Fenugreek s shoot DISCUSSION Figure 10: Effect of high-nutrient deficiency on Mlondildehyde of Fenugreek s root In this study, root nd shoot incresed in plnts with mgnesium deficiency. Also dry nd fresh weight incresed in roots nd shoots. Reltive wter content in mgnesium deficiency plnts hd no significnt difference compred to control (P <0.05). Jones nd et l (1996) found tht nitrogen nd mgnesium re necessry in formtion of chlorophyll. Mgnesium trnsfers phosphorus to seed of plnts nd lso enled dehydrogense nd decroxylse enzymes nd it is lso importnt in cellulr respirtion. This element enled enzymes in oxidtion nd reduction in plnts nd it seprtes oxygen molecules in photosynthesis. In this study, in plnts with nitrogen deficiency, dry weight of roots nd shoots hd significnt decrese compred with controls. Root fresh weight incresed in plnts with nitrogen deficiency ut shoot fresh weight reduced compred with controls. Reltive wter content incresed compred to control in nitrogen deficit. Gyne G (1979), found tht with development deficiency on citrus stem elongtion lmost stopped. As result, nitrogen deficiency led to decrese dry in ll seedlings nd ll prts mker (rnches nd leves, stems, roots).with nitrogen deficiency, growth ws stopped in 76% of ll the seedlings nd smll leves ppered in complete nutrition in the control group. In this study, nitrogen deficiency led to reducing growth fctors. Alizdeh et l (2008) found tht incresing nitrogen fertilizer leds to incresed dry mtter in shoot nd root length in corn wheres with consume more nitrogen root dry mtter decresed. Jeyhooni (2009) expressed tht nitrogen deficiency led to the stoppge of growth in tomto shoots. Nitrogen excused 2 to 5 percent of dry nd ecuse nitrogen interferes in moleculr structure of chlorophyll, there is significnt positive reltionship etween nitrogen content of leves nd the

8 Ftemeh nd Prvneh / Greener Journl of Agriculturl Sciences 301 mount of chlorophyll (Cssmn et l, 1994).Rnge of chloric in tretments of nitrogen nd mgnesium deficiency is less thn chloric in tretments of phosphorus, potssium nd clcium deficiency (Mrschner, 1995).After nitrogen, phosphorus is the most importnt element of plnt nd its vilility, rises s limiting fctor in the production of griculture products worldwide.(afif et l, 1993). An importnt sign of phosphorus deficiency is drft nd short plnts. Often, phosphorus deficient plnts re confused with young plnts nd in vein re lttice nd rown in older leves in deficiency conditions (Slim, 2007). In this study, it ws oserved tht plnt growth reduced throughout the shoots nd roots. Also plnt height ws higher in control thn nitrogen deficient plnts in greenhouse. One reson for the low high in plnts under phosphorus deficiency could e due to the phosphorus effect in grin growth period. High in ll micronutrient deficiency plnts ws lower compred to control. Also in nitrogen, high ws reduced gretly. The lrgest decrese ws in growth nd dry weight of nitrogen tretment. Phosphorus is one of the key elements which ply n importnt tsk in plnts. This element hs role in energy trnsfer in plnt metolic processes, cell division, phospholipids uilding wlls of plnt cells, reproductive prts of plnts, growth nd development of lterl roots nd hir nd in formtion nd trnsport like sugrs nd strches in plnt (Bisws nd Mukherjee,1991; Bennett,1996. Hvlin et l.,1999mrschner,2002).in this study, we lso otined similr results. In this study, reltive wter content incresed in plnts deficient in potssium thn controls. 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9 302 Ftemeh nd Prvneh / Greener Journl of Agriculturl Sciences in Bhi grss (Psplumnottum Flugge). Remote Sens. Environ. 84, , 2003 Smrt, R.E. nd G.E. Binghm.(1974). Rpid estimtes of reltive wter content. Plnt Physiol., 53: Stress. Plnt Cell Physiol., 40: Toms, A., nd Cul A. (2004). Chnges in spectrl relectnce of whet leves in response to specific mcronutrient deficiency, Received 10 Octoer 2003; received in revised form 17 Septemer 2004; ccepted 17 Septemer 2004 Wng J, Zhng H, Allen RD. (2000). Over expression of n Aridopsis peroxisoml scorte gene increse protection ginst oxidtive. J. Bio & Agri., 24: Cite this Article: Ftemeh Rezie Kinji, Prvneh Rhdri (2012). Shortge on (Mgnesium nd Clcium) Dry Weight, Fresh Weight, Root nd Shoot Length, Lef Reltive Wter Content (RWC), Chlorophyll Content nd Mlondildehyde Activity in Fenugreek (Trigonell Foenum Grceum). Greener Journl of Agriculturl Sciences, 2(7): ,