Effects of different fruit juices used as carbon source on cucumber seedling under in-vitro cultures

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1 Afrin Journl of Biothnology Vol. (3), pp. 7-7, July, Avill onlin t DOI:.97/AJB.3 ISSN 3 Ami Journls Full Lngth Rsrh Ppr Effts of iffrnt fruit juis us s ron sour on uumr sling unr in-vitro ulturs Ikrm-ul-Hq*, Zr Arsh, Tsr, Ghulm Mustf, Muksh, Muhmm Umr Dhot, Sji Ali n Munzz Institut of Biothnology n Gnti Enginring (IBGE), Univrsity of Sinh, Jmshoro, 7, Pkistn. Apt 3 Jun, In this stuy, th fft of vrious ommril fruit juis (us s plnt ron sour) ws ssss on uumr (Cuumis stivus L.) v. Liz t sling stg unr spti onitions. Ss wr grmint on ½MS mium (within -ys) unr rk onitions. Thy wr su-ultur on MS (MS sl slts) mium n its rivtivs [MS, supplmnt with fruit juis in pl of suros (3%) suh s, MS (orng), MS (ppl), MS 3 (r grps) n MS (strwrry)] for six wks. Mximum sling growth ws osrv in MS n MS 3 ulturs (p >.). Signifint inrs in prolin n ruing sugrs ws msur in MS n MS in omprison to MS (ontrol) ultur. Aunn of hlorophyll ontnts (Chl, n ) n totl rotnois inluing its prursor (lyopn) ws lso foun in MS mium. Both orng n grps suppli ulturs wr most fftiv in rltion to ll growth rlt prmtrs. Ky wors: Cuumis stivus L., sling growth, prolin ontnts, totl rotnois, lyopns. INTRODUCTION Cuumr (Cuumis stivus L.) is tnr hrous nnul vgtl vin rop us for its frsh fruit ssrts. In ition to liious tst, it is firly unnt with loris n high miinl vlus for humn ings. This rop n srv s n tiv rug for srting n promoting flow of urin s wll s usful for oth high n low loo prssurs u to high ontnts of potssium (. to. mg g - ) in fruit (Kns, 979). In Pkistn, its totl proution is 7 tons (Anon, ). Aoring to ths figurs, proution is quit low. Inrs in uumr proution lik tht of ny othr rop, n hiv ithr y ringing mor r unr ultivtion or y opting improv vritis n ttr ulturl prtis (Wsm t l., ). Bus of *Corrsponing uthor. E-mil: ro.ikrm@yhoo.om. Tl: Arvitions: MS, Murshig n Skoog; B Vit, vitmins B ; Chl, hlorophyll; F Wt, frsh wight; D Wt, ry wight; NOCl, soium hypohlri. nrrow gnti sis, ross hyriiztion is vry iffiult twn its losly rlt spis (Dn n Custrs, 99). Aspti improvmnt ginst surf growing pthogns is possil. A rprouil tissu ultur systm of plnt ithr vlop from s or rgnrtion of omplt plnt tht hs to improv for this rop (Gmorg t l., 9). Normlly, plnt growth rquirs rohyrt s sour of fr nrgy to run its ll growth prosss. Its ontinuous supply is ssntil us in vitro ulturs hv slow noli prosss with limit light intnsity, gs xhng rts n with high rltiv humiity. Suros is most wily us ron sour in plnt tissu ultur. It lso ts s n osmoti gnt to support th growth of ultur tissus (Alkht, ) Fruits r rih sour of sugrs n vitmins. Ths omponnts my lso usful for plnt tissu growth. Thir us in nutrint mium not only nhns growth rt, ut lso triggrs iffrntition in ultur tissus (Kinnrsly n Hnrson, 9: Siiqu n Pswn, 99; Wsm t l., ). Fruit juis lso ontin som growth inhiitors suh s thyln n ABA tht oul slow own growth rt ut mort th ffts of

2 Ikrm-ul-Hq t l. 7 Tl. Plnt nutrint mium with omposition of iffrnt juis us for th stuy of thir ffts on sling growth in uumr (C. stivus L.) v., Liz (-wks ultur). Mium Mium omposition Jui omposition (. g D wt) MS MS sl mium (ontrol) (Murshig n Skoog (9) ½MS Hlf onntrtion of MS sl slts (MS ) MS MS + ml orng jui (ontin 3% sugr).3% vit A,.% vit C,.% rioflvin, 3.% niin,.9% C,.3% F, 3.% vit B,.% folt, 3.% P,.3% Mg, 3.% pntthnt MS MS + 3 ml ppl jui (ontin 3% sugr) % sugrs, 7% frutos,.7% suros, % gluos,.7% mli i,.% ptin,.% strh,.% polyphnols,. protins,.% vitmins (minly sori i),.% shs,.3% β-rotn. MS 3 MS + ml r-grps jui (ontin 3% sugr) % rohyrts,.7% nitrt,.% protin,.% fr mino is,.% humin,.% Al,.7% B,.% C,.% Cl,.3% Cu,.3% F,.% Mg,.% Mn,.% K,.% P,.% R,.% Si,.% N,.3% SO. MS MS + 9 ml strwrry jui (ontin 3% sugr).% protin,.% ft,.% rohyrt,.% fir,.% sugr,.% C,.% F, Mg,.% P,.% N, % K,.7% Zn,.% Cu,.% vit C,.% thimin,.3% rioflvin,.% niin,.% vit B,.7 vit A,.% vit E,.% vit K. growth nhnrs (uxins n ytokinins) in in vitro ultur (Als, 973; Mttoo n Suttl, 99; Pu n Chi, 993). Silvr nitrt (AgNO 3 ) n oltous ions grtly improv plnt rgnrtion ffiiny in oth monots n iots (Lu n Yng, 97; Girihr t l., 3). Plnt vlopmnt is ompromis on vrious growth stgs n s grmintion, n its mrgn stgs r mor snsitiv. Th im of this stuy ws to hrtriz th growth promoting tivity of vrious juis on uumr sling unr in-vitro y rpling sugr of nutrint mium. MATERIALS AND METHODS Hlthy ss of uumr (C. stivus L.) v., Liz wr slt n wsh thoroughly with running tp-wtr. Thy wr isinft for surf growing miros with 3% Roin Blh ( % NOCl). Aftr min, ss wr rins for 3 tims with striliz H O. Striliz ss wr sok in striliz H O for h, thn ultur on ½MS [MS (Murshig n Skoog, 9) sl slts with B vit omplxs n 3% sugr] mium for ys in th rk. Grmint ss wr su-ultur on fiv iffrnt plnt nutrint mium n inut unr light onitions (Tl ). Almost -wks ol slings wr rmov from mium n ln from gr y wshing with tp wtr. Thir iomss ws msur in orr to wigh thir frsh wight (F Wt), whil thir ry wight (D Wt) ws msur y rying t 7 C for 3-ys. Chlorophyll ontnts n totl rotnois wr nlys from frsh lvs y grining in % ton (Arnon, 99; Ngt n Ymshit, 99). Crohyrts wr xtrt from plnt ry mtrils oring to Cih n Brun (97) y homogniztion in ml sugrs xtrtion uffr (glil ti i: mthnol: wtr, ::, v/v/v). Thy wr quntifi following Duois t l. (9) mtho. Ruing sugrs wr lso trmin y following Millr (99) n totl protins oring to Brfor (97) mthos. Nitrt ontnts wr trmin y using Morris n Rily (93) mtho. Asort (Shlt n Numnn, ), prolin (Bts t l., 973) n phnoli (Ozyigit t l., 7) ontnts wr lso trmin. Th signifin of th ollt vlus from h ultur (with rplits) ws lult using COSTAT omputr pkg (CoHort softwr, Brkly, USA). RESULTS AND DISCUSSION An xprimnt ws prform y ulturing grmint ss (-ys ol) of uumr (C. stius L.) v., Liz on MS ulturs supplmnt with vrious fruit juis (Tl ). Diffrnt fruit juis wr in th MS

3 A B C D E Figur. Dvlopmnt of slings of uumr (C. stivus L.) v., Liz on iffrnt plnt nutrint sl mium [Murshig n Skoog, (9) sl slts] suppli with iffrnt juis in pl of ron sour unr in-vitro onition ( wks ultur). : Growing sling on simpl MS mium; : sling on simpl MS mium suppli with orng jui in pl of 3% sugr; : sling on MS mium suppli with ppl jui inst of 3% sugr; : sling growing on MS plnt nutrint mium suppli with r-grps jui in pl of 3% sugr n : slings growing on MS plnt nutrint mium supplmnt with strwrry jui in pl of 3% sugr. mium just to ring th lvl of sugr up to 3% (MS without sugr), whih is rquir for plnt growth unr in-vitro onitions. Ths juis oul lso sour of vrious nutrint sustns, whil this xprimnt ws fous on sugr ontnts only. Ths ulturs wr mintin in light onitions for -wks. Diffrnt rsponss of growing slings wr osrv from ontrol to jui suppli nutrint ulturs (Figurs n ). Signifint hng in growth ws osrv mong th jui ulturs. Th plnt iomss of th uumr slings ws inrs (p >.). Th most fftiv plnt growth ulturs wr MS (orng) n MS 3 (grps). Mximum plnt hight n root lngth ws osrv in MS n MS 3 ulturs. Ths two ulturs wr onsir to nhn th vgttiv plnt growth of th slings signifintly. Mnwhil, shoot ry wight ws highr in MS n root ry wight in MS (ppl), not in MS 3 (Figur ). Th prformn of th morphologil hrtrs ws not fft vrsly us of th pplition of fruit juis in MS mium thn ontrol (MS suppli with 3% sugr) ulturs. Supplmnt of juis nhn th growth rt of th ultur slings in th form of plnt hight, root lngth n plnt iomss (Figurs n ). Suh lrt growth rt sms to triggr y th pplitions of hormons in th ulturs (Klly t l., 97; Blstri t l., 99; Ry t l., ; Puhoo n Rmurn, ). Chlorophyll ontnts wr lso fft signifintly. Chlorophyll (Chl ) ws highr in MS ulturs ut quntifi lmost th sm in MS n MS (strwrry) ulturs. Similrly, mximum Chl ws lso osrv in MS ulturs. Lyopns wr msur highr in MS n MS ulturs, whil totl rotnois wr highr in MS n MS ulturs. Biosynthsis of lyopn ws sustntilly lowr mong th ulturs us it is prursor (yli) of rotnois. Som xtrnl strsss on th growing plnts us n inrs in totl rotnois (Snmnn, ; Mrov t l., ). Mnwhil, th MS ultur rprsnt th st iosynthsis of ll plstis ut h rs iffrntilly mong othr fruit jui suppli ulturs. Minimum plstis wr osrv in MS n MS. It mns tht oth wr involv in th inhiition of thir iosynthsis. Suh inhiitions r th prim rols of spti ulturing to stop or ru th iffrntition prosss of th vloping tissus. Totl rotnois r lso th sign of growth inhiition u to rtin ppli strsss ut its mximum onntrtion ws osrv in orng jui suppli ulturs. Thr sling growth rt ws vry ffiint thn othr ulturs. Th influning ftors of th rotnois r not lr. Signifint inrs in totl protins, sugrs n ruing sugr ontnts ws osrv in th slings ultur on MS 3 mium. Eh rs in MS s wll s in MS ulturs. Similrly, protin ontnts wr highr in MS, whil phnolis n nitrts in MS. Mximum sort ontnts wr msur in MS ulturs. Almost h prmtr ws prform iffrntly in th prsn of vrious fruit juis in th ulturs (Figur ). During slings growth, umultion of prolin n ruing sugrs hs lso n onsir s sign for ioti strsss on th growing plnts or tissus in

4 Ikrm-ul-Hq t l. 77 Shoot hight (m) Root lngth (m)... Shoot F Wt (g) ** Root F Wt (g) Lf r (m)* Shoot D Wt (g) Root D Wt (g) Plstis mg/g 7 3 Chl Chl Chl Crotnois Lyopns Protins n sugrs (mg/g) 3 3 Protin Totl s ugrs Ruing s ugrs Diffrnt io-om ponnts 9 7 Prolin (mg/g) Phnolis (mm/g) Nitrt (mg/g) Asort (mg/g) 3 MS MS MS MS 3 MS MS MS MS MS 3 MS f Figur. Efft of ommrilly vill fruit jui on plnt growth rlt morphologil n io-hmil prmtrs of th wks ol sling ultur of uumr (C. stivus L.) v., Liz unr in-vitro onitions. Brs with lttrs shown t iffrnt growth prmtr r signifintly iffrnt t p<. y using th Dunn s multipl rng (DMR) tst. mium. Both of ths ontnts wr highr in MS n MS 3 ulturs. It mns tht orng n grp juis wr involv in th nhnmnt of sling growth npplition of somwht strsss. In this stuy, th pplitions of iffrnt fruit juis prform signifint rol in sling growth. Eh growth hrtr hs shown nfiil inrmnt thn ontrol ulturs. Som ulturs suppli with ppl n

5 7 Afr. J. Biothnol. strwrry juis wr not prform s goo s orng n grp. This hvior of growth my us of iffrn in jui ompositions. Som jui sustns r growth supportiv, whil othrs my not triggr som othr mo of growth. Mnwhil, thr is mor n to prform suh xprimnts for th purpos of intifying ny spifi mo of growth tht is unr ontrol of spifi jui omponnt. Orng n grp juis r st nutrint sours for uumr sling growth. Thy my lso hv th ility to prform th st in-vitro plnt mss proution or in-irt plnt rgnrtion from singl plnt ll or its tissu. REFERENCES Als FB (973) Ethyln in plnt iology. Ami prss Nw York, p 3. Alkht A (). Comprison Effts of Suros n Dt Plm Syrup on Somti Emryognsis of Dt Plm (Phonix tylifr L.). Amr. J. Biothnol. Biohm. : 9-3. Anon (). Sttistis of Pkistn, -. Govrnmnt of Pkistn, Ministry of Foo, Agriultur n Livstok Division (Eonomis Wing). Islm. Arnon DT (99). Coppr nzyms in isolt hloroplsts polyphnol oxis in Bt Vulgris. Plnt Physiol. : -. Blstri E, Pizzi L, Cinlli F (99). Survivl n growth of trnsplnt n nturl slings of Posioni oni (L.) Dlil in mg ostl r. J. Exp. Mr. Biol. Eol. : 9-. Bts LS, Wlrn RP, Tr ID (973). Rpi trmintion of fr prolin for wtr strss stuis. Plnt Soil, 39: -7. Brfor MN (97). A rpi n snsitiv mtho for th quntittion of mirogrm quntitis of protin utilizing th prinipl of protin-y ining. Anl. Biohm. 7: -. Cih AJ, Brun WA (97). Efft of po rmovl on nonstruturl rohyrt onntrtion in soyn tissu. Crop Si. : Dn NAPM, Custrs JBM (99). Introution rsistn into uumrs y intrspifi hyriiztion. Crop Imp. Prot. pp Duois M, Gills KA, Hmilton JK, Rrs PA, Smith F (9). Colorimtri mtho for trmintion of sugrs n rlt sustns. Anl. Chm. : 3-3. Gmorg OL, Dvis P, Sthlut RW (9). Somti mryognsis in ll suspnsion ulturs of glyin spis. Plnt Cll Rp. : 9-. Gmorg O, Millr R, Ojim K (9). Nutrint rquirmnts of suspnsion ulturs of soyn root lls. Exp. Cll Rs. : -. Girihr P, Inu EP, Rmu DV, Rvishnkr GA (3). Efft of silvr nitrt on in vitro shoot growth of Coff. Trop. Si. 3: -. Kns JM (979). Enylopi of Mil Foos. Thorns Pu Lt, Willing Borough, North Ampotnshin UJ, p. 9. Klly JA, Fuss CM, Hll JR (97). Th trnsplnting n survivl of turtl grss Thlssi tstuinum, in Bo Cig By, Flori. Fish. Bull. USA. 9: 73-. Kinnrsly AM, Hnrson WE (9). Altrntiv rohyrts promot iffrntition of plnt lls. Plnt Cll Tiss. Org. Cult. : 3-. Lu OL, Yng SF (97). Inhiition of thyln proution y oltous ion. Plnt Physiol. : -7. Mrov I, Brirov E, Koi R, Fril Z, Slovk B, Pokorn J (). Influn of xognous strss ftors on proution of rotnois y som strins of rotnogni ysts. Ann. Miroiol. : 73-. Mttoo AK, Suttl JC (99). Th plnt hormon thyln. CRC prss. Bo Rton, Flori, p Millr GL (99). Us of initrosliyli i rgnt for th trmintion of ruing sugr. Anl. Chm. 3: -9. Morris AW, Rily JP (93). Th trmintion of nitrt in s wtr. Anl. Chm. 9: Murshig T, Skoog F (9). A rvis mium for rpi growth n iossys with too tissu ultur. Physiol. Plnt, : Ngt M, Ymshit I (99). Simpl mtho for simultnous trmintion of hlorophyll n rotnios in tomto fruit. J. Jpn. So. Foo Si. Thnol. 39: 9-9. Ozyigit II, Khrmn MV, Ern O (7). Rltion twn xplnt g, totl phnols n rgnrtion rspons in tissu ultur otton (Gossypium hirsutum L.). Afr. J. Biothnol. : 3-. Pu EC, Chi GL (993). D novo shoot morphognsis n plnt growth of mustr (Brssi jun) in vitro in rltion to thyln. Physiol. Plnt, : 7-7. Puhoo D, Rmurn RA (). Stuy on th us of rrot jui in th tissu ultur of Duus rot. Afr. J. Biothnol. 3: -. Ry CS, Bhnj MR, Rju VS (). Cssi uniflor Millr: nw ror for Anhr Prsh, Ini. In. J. Forst, 3: 3-3. Ry KR, Hogs HF, Kimll BA (). Crop osystm rsponss to glol hng: Cotton, In KR Ry, HF Hogs (.) Climt hng n glol rop proutivity. CABI Pul., Nw York. pp Snmnn G (). Gnti mnipultion of rotnoi iosynthsis: strtgis, prolms n hivmnts. Trn Plnt Si. : -7. Shlt A, Numnn PM (). Exognous sori i (Vitmin C) inrss rsistn to slt strss n rus lipi proxition. J. Exp. Bot. : 7-. Siiqu AB, Pswn L (99). Efft of growth rgultors n orgni supplmnts on iffrntition of Cymiium longifolium P., in vitro. J. Hill Rs. : 3-3. Wsm K, Kmrn MO, Jiln SM (). Efft of nitrogn on growth n yil of uumr (Cuumis stivus L.). J. Agri. Rs. : 9-.