CONCENTATION OF MINERAL ELEMENTS IN CALLUS TISSUE CULTURE OF SOME SUNFLOWER INBRED LINES

CONCENTATION OF MINERAL ELEMENTS IN CALLUS TISSUE CULTURE OF SOME SUNFLOWER INBRED LINES M. Sri 1, Drgn Vsi, Lj. Vsiljevi, D. Skori, Snezn Mezei nd Sloodnk Pjevi 2 ABSTRACT Conentrtion of minerl elements in lli of six sunflower inred lines hve een determined. Clli were indued from otyledons of seven dys old seedlings grown under sterile onditions. The differenes etween inred lines in onentrtion of N, P, K, Mg, Fe nd Mn were signifint, wheres differenes in the onentrtion of C, N, Cu nd Zn were insignifint. Averge onentrtions of minerl elements in lli were lower thn verge onentrtions of minerl elements in leves of plnts grown in vivo. Key words: minerl elements, llus, sunflower, inred lines. N INTRODUCTION umerous riteri hve een used to evlute the otined effets of geneti speifiity of the minerl nutrition in plnts. It should e emphsized tht in previous investigtions the onentrtion nd ontent of minerl elements in plnts hve een most frequently used s riterion for the ourrene of geneti speifiity. Cultivr speifiity of minerl nutritions is mnifested through the differenes in ion uptke, their trnsport, distriution nd umultion in individul orgns, s well s in their reutiliztion, efflux, metoli role nd iomss prodution. The differenes mentioned re found in plnt orgns of different ge of oth ultivted plnts nd those inhiting nturl phytoenoses. All differenes re found under oth rtifiil nd nturl onditions. Numerous results, ited in reviews (Epstein, 1972; Sri, 1981, 1983; Clrk, 1983), show tht there is differene etween genotypes of different plnt speies onerning onentrtion of some elements of minerl nutrition of plnts grown in vivo. A review of results onerning sunflower (Helinthus nnuus L.) is given the pper of Sri et l. (1991). Prolems of geneti speifiity of minerl nutrition of plnts grown in vitro were investigted to lesser extent. One must keep in mind the ft tht physiology nd iohemistry of plnts grown in vitro differ from those grown in vivo. Here is one exmple. The intensity of growth nd development of plnts grown in vivo is funtion of net photosynthesis whih, most often, orreltes with the intensity of photosynthesis nd the lef re. In vitro, the growth is funtion of minerl elements nd orgni omponents of the medium, euse the intensity of photosynthesis nd lef re re negligile. Physiologil nd iohemil differenes re fully expressed in llus ultures. The growth nd morphogenesis of plnts grown in vitro ould e effetively influened y optiml onentrtion of the elements of minerl nutrition nd other orgni omponents. The suess of growing plnts in vitro is relted primrily to the omposition of the medium. Still, the numer of ppers deling with prolems of geneti speifiity of minerl nutrition of plnts grown in vitro is onsiderly sprse. The review of ppers deling with genetil spet of minerl nutrition of plnts grown in vitro is given in pper of Sri et l. (1995). We elieve tht the results referring to minerl nutrition of plnts grown in vitro will ontriute not only to improving this method, ut lso to the further progress in solving the prolems of geneti speifiity of minerl nutrition of plnts nd even the ion uptke mehnism. MATERIALS AND METHODS Six sunflower inred lines,, PH- BC 2-152A,,, nd were used in the experiment. Eh inred line ws represented y ten Institute of Field nd Vegetle Crops, M.Gorkog 30, 21000 Novi Sd, Yugoslvi 1 Serin Ademy of Sienes nd Arts, Knez Mihjlov 35, 11000 Belgrde, Yugoslvi 2 Fulty of Nturl Sienes, Trg Dositej Ordovi 7, 21000 Novi Sd, Yugoslvi 37

38 ROMANIAN AGRICULTURAL RESEARCH Numer 7-8 / 1997 plnts, whih were donors of the initil explnts. Clli were indued from otyledons, tken from seven dys old sunflower seedlings grown in sterile environment (Vsiljevi nd Vsi, 1994). The explnts were ultured on the MS medium (Murshige nd Skoog, 1962), supplemented with 1 mll -1 enzylminopurine nd 2 mll -1 nphtleneeti id. After three weeks of ulture lli were ut in two prts. Mss of fresh nd dry mtter of one prt of eh llus ws mesured. The other prt of eh llus ws pled on fresh medium nd grown under fully ontrolled temperture (25 0 C), photoperiod (16:8; light:drk) nd humidity (85%). The sme proedure hs een repeted for five times. After tht, lli were rinsed with distilled wter, dried to onstnt ir-dry mss, merted nd prepred for hemil nlyses. Clli were nlysed for nitrogen ontent y the Kjeldl method, phosphorus ontent y molydte-vndte spetrophotometry, potssium ontent y flme photometry, lium, mngnese, sodium, iron, zin, mgnesium nd opper ontent y tomi sorption spetrosopy. Dt were proessed y the nlysis of vrine. RESULTS AND DISCUSSIONS The differenes in the onentrtion of minerl elements in the lli otined from different inred lines vried from highly signifint to insignifint (Tles 1 nd 2). Avrge onentrtion of minerl elements in lli were ompred to verge onentrtions of minerl elements in root, stem nd leves of intt plnts (Sri et l., 1991) (Figures 1, 2 nd 3). Figure 1. Averge onentrtions of some minerl elements in lli nd root of sunflower inreds Tle 1. Conentrtion of some mroelemts in lli of six sunflower inreds (mg/100 g dry mtter) Genotype N*** P** K* C ns N ns Mg*** 3197 281 2833 825 700 96 4669 331 24 808 767 89 5222 350 19 841 750 90 5534 362 2783 725 767 87 5282 315 2583 816 767 91 4828 332 2833 733 650 99 Averge 4789 328 2561 792 733 90 LSD 5% 1% 218 306 31 44 511 716 282 89 125 1.7 2.4 Tle 2. Conentrtion of some miroelemnts in lli of six sunflower inreds (ppm) Genotype Fe*** Mn*** Zn ns Cu ns 225 278 285 315 212 8 193 167 148 165 190 198 192 18 20 13 15 Averge 268 7 LSD 5% 1% 32 45 12 36 51 5.5 7.8

M.SARIC ET AL.: CONCENTRATION OF MINERAL ELEMENTS IN CALLUS TISSUE CULTURE OF SOME SUNFLOWER INBRED LINES 39 Figure 2. Averge onentrtions of some minerl elements in lli nd stem of sunflower inreds Figure 3. Averge onentrtions of some minerl elements in lli nd leves of sunflower inreds Nitrogen Nitrogen onentrtion in lli rnged from 5534 () to 3197 mg/100 g of dry mtter (). Averge nitrogen onentrtion of ll six inred lines ws 4789 mg/100 g of dry mtter. Averge nitrogen ontent of twenty sunflower inreds grown in vivo depending of plnt orgn ws 6765 (leves), 3462 (stem) nd 3829 (root) mg/100 g of dry mtter. On the sis of the otined results it ould e onluded tht nitrogen onentrtion in lli of inred lines ws within the limits of plnts grown in vivo. Phosphorus Phosphorus ontent vried s well, nd the order of miniml nd mximl onentrtions ws the sme s for nitrogen, i.e. hd the lowest (281 mg/100 g of dry mtter) nd the highest phosphorus onentrtion (362 mg/100 g of dry mtter). In omprison to phosphorus onentrtion in intt plnts, the verge phosphorus onentrtion ws signifintly lower in lli (328 mg/100 g of dry mtter), euse it rnged from 730 to 1687 mg/100 g of dry mtter in leves of intt plnts. Potssium Potssium onentrtion ws the highest in (2833 mg/100g of dry mtter), opposite to nitrgen nd phosphorus onentrtion. The lowest onentrtion ws 19 mg/100 g of dry mtter in. In omprison to intt plnts where onentrtion vried from 3900 to 6300 mg/100 g of dry mtter it ould e sid tht potssium onentrtion ws more thn three times higher in intt plnts thn in lli. Clium The differenes in lium onentrtion in lli of tested inred lines were insignifint. If we ompre lium onentrtion in in vitro nd in vivo grown plnts, lium onentrtion in lli ws signifintly lower thn in intt plnts. Sodium The differenes in sodium onentrtion in different inreds were insignifint. Mgnesium The inred lines tested differ signifintly in mgnesium onentrtion. Inred line hd the highest (96 mg/100 g of dry mtter) nd the lowest mgnesium onentrtion (87 mg/100 g dry mtter). Mgnesium onentrtion in plnts grown in vivo rnged from 563 to 822 mg/100 g dry mtter while it ws five times lower in lli. Iron Iron onentrtion ws signifintly different in different inreds nd hd the lowest (225 ppm) nd the highest iron onentrtion (315 ppm) Mngnese The differenes in mngnese onentrtion in tested inred lines were signifint hd the highest (212 ppm) nd the lowest mngnese onentrtion ( ppm). The differenes in the onentrtion of zin s well s opper were insignifint. Clli ould e onsidered onvenient for studying the geneti speifiity of minerl nutrition euse mny ftors tht use signifint vriility of results in vivo re negligile in vitro.

40 ROMANIAN AGRICULTURAL RESEARCH Numer 7-8 / 1997 A llus is generlly more homogenous thn n intt plnt, lthough it is not idelly homogeneous. Its diversity is mnifested in the totl numer of ells, numer of meristemti ells, ell size, differentition of somti ells, nd vuole size. Division tivity of ells is not uniformly rrnged throughout llus tissue. It is well known tht in in vitro ulture new vriility n pper in signifintly higher perentge thn spontneous muttions. This vriility is lled somlonl vriility nd n e oserved on morphologil, ytologil nd moleulr level. In our experiment, we wnted to hieve the highest possile geneti uniformity within eh tested genotype nd to void differenes in nutrient onentrtions whih ould our due to geneti divergene within the tested genotypes, so we hve used different inred lines s donors of explnts. Differenes in onentrtion of minerl elements were otined etween different sugreet ultivrs (Mezei et l., 1995). Averge onentrtion of minerl elements in lli ws lower thn in intt plnts lthough they were grown on medium supplemented with higher onentrtions of nutrients thn intt plnts. It ould e explined y the ft tht reltive to their totl fresh or dry weight, rooted plnts in vivo hve, through root hirs nd myorrizl fungi, very lrge surfe re involved in nutrient uptke (Mengel, 1984) ompred with lli. Also, in in vivo nutrient onentrtions in medium re kept onstt wheres in in vitro medium the initil onentrtions of nutrients re very high, ut then derese rpidly (Leifert et l., 1995). Averge onentrtions of minerl elements in lli hve een ompred to verge onentrtions of minerl elements in root, stem nd leves of intt plnts. Generlly, if we ompre onentrtions of tested minerl elements in intt plnts nd in lli, we n onlude tht onentrtion of ll minerl elements ws lower in lli thn in intt plnts (Sri et l., 1991), lthough lli were grown on medium supplemented with higher onentrtions of minerl elements thn intt plnts (Sri et l., 1995). It would e interesting to investigte the retions of plnts of the sme genotype ut of different origin to individul minerl elements. REFERENCES Clrk, R.B., 1983. Plnt genotype differenes in the uptke, trnslotion, umultion nd use of minerl elements required for plnt growth. In: Geneti Aspets of Plnt Nutrition. Mrtins Nijhoff/J.W. Junk Pulishers, the Hgue/Boston/Lnster: 49-70 Epstein, E., 1972. Physiologil genetis of plnt nutrition. In: Minerl Nutrition of Plnts: Priniples nd Perspetives. John Wiley nd Sons, New York: 325-344. Leifert, C., Murphy, K.P., Lumsden, P., 1995. Minerl nd rohydrte nutrition of plnt ell nd tissue ultures. Crit.Rev.Plnt Si. 14(2): 83-109. Mengel, K., 1984. Ernhrung und Stoffwehsel der Pflnze. Gustv Fisher Verlg. Stuttgrt, Germny. Mezei, S., Sri, M., Pjevi, S., Kovev, L., 1995. Conentrtion of minerl elements in llus tissue ultures of two sugreet (Bet vulgris L.) ultivrs. Arh.Biol.Si., Belgrde, 47 (1-2): 29-32. Murshige, T., Skoog, F., 1962. A revised medium for rpid growth nd iossys with too tissue ulture. Physiol. Plnt 15: 473-497. Sri, M., 1981. Geneti speifiity in reltion to plnt minerl nutrition. J. Plnt Nutr. 3: 743-764. Sri, M., 1983. Geneti improvement of rops yields s relted to plnt nutrient requirements. 9th CIEC World Fertilizer Congress. Budpest, Hungry: 115-128. Sri, M., Krsti, B., Skori, D., 1991. Element diversity in sunflower inred lines. Heli 14 (15): 41-48. Sri, M., Mezei, S., Ruzi, D., 1995. Geneti spets of minerl nutrition of plnts grown in vitro. Arh.Biol.Si., Belgrde 47(1-2): 1-12. Vsiljevi, Lj., Vsi, D., 1994. Plnt regenertion of sunflower (Helinthus nnuus L.) in vitro. Pro. Oil nd Protein Crops Meeting of EUCARPIA. Alen, Bulgri: 182-184.

M.SARIC ET AL.: CONCENTRATION OF MINERAL ELEMENTS IN CALLUS TISSUE CULTURE OF SOME SUNFLOWER INBRED LINES Tle 1. Conentrtion of some mroelemts in lli of six sunflower inreds (mg/100 g dry mtter) Genotype N*** P*** K* C ns N ns Mg*** 3197 4669 5222 5534 5282 4828 281 331 350 362 315 332 2833 24 19 2783 2583 2833 825 808 841 725 816 733 700 767 750 767 767 650 96 89 90 87 91 99 Averge 4789 328 2561 792 733 90 5% 218 31 511 89 1.7 LSD 1% 306 44 716 282 125 2.4 41 Tle 2. Conentrtion of some miroelemnts in lli of six sunflower inreds (ppm) Genotype Fe*** Mn*** Zn ns Cu ns 225 278 285 315 212 8 193 167 148 165 190 198 192 Averge 268 7 5% LSD 1% 32 45 12 36 51 5.5 7.8 18 20 13 15 Figure 1. Averge onentrtions of some minerl elements in lli nd root of sunflower inreds

42 ROMANIAN AGRICULTURAL RESEARCH Numer 7-8 / 1997 Figure 2. Averge onentrtions of some minerl elements in lli nd stem of sunflower inreds Figure 3. Averge onentrtions of some minerl elements in lli nd leves of sunflower inreds