Pálfi, Gábor

English

Pálfi Gábor

University of Szeged

A N E W I N D I C A T O R O F A B N O R M A L A M I N O A C I D A N D P R O T E I N M E T A B O L I S M I N D I S E A S E D P L A N T S : R I C E , P O T A T O , S O Y B E A N , T O B A C C O G . P Á L F I Ins t i tu te for P l an t Phys io logy , At t i l a József Univers i ty , Szeged (Received September 26, 1966) I n t r o d u c t i o n T h e f r ee amino acid con ten t of p l an t organisms m a y be increased by means of a large N or P supply , or in consequence of cer ta in diseases ( B I D W E L L et al . , 1 9 6 4 ; F A R K A S and K I R Á L Y , 1 9 6 1 ) . By p a p e r c h r o m a t o g r a p h y w e h a v e looked for a physiological indicat ion to d e m o n s t r a t e beyond any d o u b t if o r no t the increase of t h e f ree a m i n o acid con ten t is a consequence of some in fec t ion . A n u n k n o w n substance, lying a t 0,64 Rf be tween the spot of / - a m i n o bu ty r ic acid and val ine , was detected f i rs t in the c h r o m a t o g r a m of rice ( P Á L F I , 1 9 6 4 ) . F ixed wi th copper n i t ra te , this substance, in dis t inct ion to a m i n o acids, gives a blue co lour instead of a red one. The re fo re , the u n k n o w n substance has been designated by us as „b lue s u b t a n c e " a n d its indicat ion as the „b lue tes t . " In the case of rice, it a p p e a r e d t h a t u n d e r condi t ions p romot ing the occurence of diseases ( P Á L F I , 1 9 5 6 ) o r even p r o v o k i n g them ar t i f ic ia l ly ( P O D H R A D S Z K Y , 1961) the m o r e resistant fo re ign variet ies con ta ined less of the „b lue subs tance" t han the less res is tant H u n g a r i a n types . The re fo re , the test m a y be suggested as a physiological indica t ion to select disease-resistant variet ies . B y p repa r ing an a m i n o ac id - f r ee pro te in ex t rac t of diseased leaves and hydro l i z ing it w i th hyd roch lo r i c acid, the „b lue subs tance" c a n n o t be f o u n d a m o n g t h e a m i n o acids p roduced . Thus it is not a componen t of pro te in . When the f r ee a m i n o acid ex t rac ts con ta in ig the „b lue subs tance" a re h y d r o l i z e d , all the amides a n d pept ides in t hem decompose bu t the „blue subs tance" remains . Thus it is nei ther an a m i d e nor a pept ide . Since the „b lue subs tance" has been detected in whea t , as well ( P Á L F I , 1 9 6 5 ) , ou r invest igat ions were ex tended to o the r p lants , too. Mater ia ls and m e t h o d s Ascending one-d imens iona l pape r s (WHATMAN N o 1) have been slowly deve loped by being cooled in a 2 :1 :1 solvent of bu tano l -ace t i c a c id -wa te r . In the second dimension a pheno l -water mix tu re (4:1) has been app l ied . Final deve lopmen t has been secured by n inhyd r ine at 98 ° C . T h e alcohol ic ex t r ac t of 1 g f resh substance or 200 mg substance dr ied at 65 °C served as s t a r t ing ma te r i a l . Leaves of hea l thy and diseased p lan ts were collected f r o m t h e 72 G. PALF1 same parcels d u r i n g f lowering. 12 H u n g a r i a n a n d fore ign rice var ie t ies were s tudied a m o n g o thers the Hungarian Dungban sbali. P o t a t o was f r o m the sor t of „Tompa rozsa"; soy-bean Jregi korona" a n d tobacco „Szabolcsi". T h e diseases were : „blast disease" a t rice (Piricularia oryzae CAV.), „leaf roll virus" (Cormm solani HOLMES) a t p o t a t o ; „soybean mosaic" at soy-bean and „ M a r m o r tabaci var. vulgare Holmes" at tobacco. Resul t s Figure 1 shows tha t the „b lue subs tance" is present besides rice in the leaf-ex t rac ts of diseased p o t a t o a n d tobacco, as well . A t the same time, t h e ex t rac t s of hea l thy p lan t s collected f r o m t h e same parcels as the diseased ones did no t con-ta in the „b lue substance" . F igure 2 i l lustrates t h a t the blue spot can be f o u n d in t h e c h r o m a t o g r a m of the leaf ex t rac t s of v i rus - in fec ted soy-bean, too. A n d the „b lue subs tance" couldn ' t be f o u n d in the case of the hea l thy con t ro l , e i ther . I t can be seen tha t „blue sbus tance" reaches the largest R f value in the second d i -mension, i. e., in pheno l -wa te r solvent . A £ C D E ? Fig. 1. C h r o m a t o g r a m of leaf ex t rac t s of heal thy a n d diseased plants . A = hea l thy r ice; B = Pi r icu lar ia infected rice; C = hea l thy p o t a t o ; D = virus infected p o t a t o ; E = hea l thy tobacco ; F = virus infected tobacco, ft = blue spot, u n k n o w n . Figure 3 shows the two-d imens iona l d e v e l o p m e n t of the leaf ex t rac t s of v i rus - in fec ted tobacco — the „b lue subs tance" a p p e a r s here, too. In this f igu re as well as in the previous ones it is obvious t h a t the „b lue subs tance" a p p e a r i n g , also asparag ine can be found in a cons iderable a m o u n t . F u r t h e r it w a s exper ienc-ed tha t i diseased p lants w h e r e / - a m i n o bu ty r i c acid was present in a larger a m o u n t , m o r e „b lue sub tance" could be detec ted . It m a y be seen in f igures 4 and 5 tha t the „b lue subs tance" a p p e a r s on ly in the th in layer c h r o m a t o g r a m of the diseased p o t a t o . a n e v t i n d i c a t o r i n d i s e a s e d p l a n t s 7 3 Fig. 2. C h r o m a t o g r a m of ex t r ac t f r o m vi rus- infec ted soybean leaf . /} = the „blue spot" . Fig. 3. C h r o m a t o g r a m of ex t r ac t f rom vi rus- infec ted tobacco leaf , rf = the „blue spot" . Fig. 4. Thin laver c h r o m a t o g r a m of hea l thy p o t a t o . 1 = Leu + l i eu ; 2 = Val + M e t ; 3 — - A m b ; 4 = Ala + T h r ; 5 = G l u — N H , - f Arg ; 6 = G l u ; 7 = Scr + G l y ; 8 Asp; 9 = Lvs ; 10 = Asp — N H , ; 11 = Cys . Fig. 5. Thin layer c h r o m a t o g r a m of diseased po ta to . 1—7. numbers as in Fig. 4; 8 = Lvs; 9 = Asp. — NH-.,; 10 = A s p ; 11 = Cys ; 6 = blue spot. From the leaves of rice con ta in ing „b lue subs tance" a t a p w a t e r ex t rac t has also been p r e p a r e d and inocula ted wi th a pure cul ture . Bacterii of seven species, a m o n g them Bac. subtilisATCC 6633 and Escherichia coli 0 111 consumed the a m i n o acids a t 30 C ° in seven days but the „blue subs tance" remained unchanged . It fo l lows f r o m this tha t this substance is no 1 - a m i n o acid a n d is not toxic. I t was detected tha t the largest amoun t of the „b lue subs tance" could be found in the leaves of p lants , the stem conta in ing less and the roots and seed the least of i t . I n the lower leaf s toreys of diseased p lan t s there was less „blue sub-s t ance" t han in the y o u n g e r u p p e r leaves. Thus the „b lue subs tance" appeared in a large a m o u n t w h e r e the pro te in metabol ism was m o r e intense. In the case of rice, dur ing r ipening of the corn when the prote in synthesis was m a r k e d l y dec-l a s s e d in the still green bu t a l r eady old uppe r leaves, too, the „blue substance" could h a r d l y be po in ted ou t f r o m the ext rac ts . These da t a indica te tha t the a p p e -arance of the „b lue subs tance" is connected wi th an increased prote in metabol i sm. 74 G. PÁLFI T h e blue spot can be detec ted not on ly in p lan t s ment ioned in this p a p e r but on the leaves of o the r diseased p lants , as well . T h u s recent ly it has been de-tected also in the c h r o m a t o g r a m s of ex t rac ts of v i rus - in fec ted bean , s u n f l o w e r and vi rus and Pbytophtora infec ted Solarium lacimatum AIT. medic ina l he rb . These p lants , howeve r , h a v e no t been studied in detai l . S u m m a r y In the course of the s tudy of the f ree a m i n o acid spec t rum of diseased rice, soybean, p o t a t o and tobacco leaves an u n k n o w n c o m p o u n d has bean de t ec t ed , the a p p e a r a n c e of which is cor re la ted wi th an a b n o r m a l physiological s t a te of t h e p lants , or w i th some d is turbances in the a m i n o acid — pro te in metabo l i sm. Refe rences B I D W E L L , G . S . , B A R B , R . A . , S T E W A R D , F . C . ( 1 9 6 4 ) : P r o t e i n s y n t h e s i s a n d t u r n - o v e r i n c u l t u r e d p l a n t t issue: Sources of carbon for synthesis a n d the f a t e of the prote in b r e a k d o w n p r o -ducts . — Na tu re . 203, 367—373. FARKAS, G . L., KIRÁLY, Z . (1961): Amide metabol ism in w h e a t leaves in fec ted w i t h stem rus t . — Physiol . P l an t . 14, 344—353. PÁLFI, G . (1964) : Einc neue, n i n h v d r i n - und isa t inposi t ive , aminosaureahn l i che V e r b i n d u n g aus Reisb la t tc rn , die das Mass der S t i cks to f fve r so rgung anzcigt . — Die N a t u r w i s s e n s c h a f t c n . 51, 489. PALFI, G . (1965): Relat ions be tween a b u n d a n t N - s u p p l y a n d the amino acid concen t r a t i on of var ious leaf levels of rice plants . — Plant and Soil. 23, 275—284. PODHRADSZKY, J . (1961): P r o v o k a t í v vizsgálati módsze rek r i z s f a j t ák P i r i cn / í j r / a - rez i sz tenc iá já -nak e lb í rá lására . — Növény te rme lé s . 10, 67—78 . PÁLFI, G . (1965): A búza t ransz lokációs aminósavai . — N ö v é n y t e r m e l é s . 14, 181 — 1 9 0 . 

A new indicator of abnormal amino acid and protein metabolism in diseased plants: rice, potato, soybean, tobacco

http://acta.bibl.u-szeged.hu/21652/1/biologica_012_fasc_003_004_071-074.pdf

A new indicator of abnormal amino acid and protein metabolism in diseased plants: rice, potato, soybean, tobacco

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