Rozniki Akdemii Rolnizej w Poznniu CCCLXXXIII (27) ALICJA SANIEWSKA, MARIAN SANIEWSKI THE EFFECT OF TRANS-2-HEXENAL AND TRANS-2-NONENAL ON THE MYCELIUM GROWTH OF PHOMA NARCISSI IN VITRO * From Reserh Institute of Pomology nd Floriulture in Skierniewie ABSTRACT. The effet of trns-2-hexenl nd trns-2-nonenl on the myelium growth of Phom nrissi on PDA medium ws investigted. Both ldehydes strongly inhiited the myelium growth of P. nrissi when pplied to PDA medium or pplied s voltiles; greter inhiitory effet ws oserved when they were pplied s voltiles. The inhiitory effet on the myelium growth of P. nrissi ws greter of trns-2-nonenl thn trns-2-hexenl. Key words: trns-2-hexenl, trns-2-nonenl, Phom nrissi, myelium growth Introdution Severl unsturted ldehydes re produed from polyunsturted ftty ids vi the lipoxygense pthwy when plnts re wounded mehnilly or infeted y pthogens. Severl of these ldehydes, inluding trns-2-hexenl nd trns-2-nonenl re found to e inhiitory to pthogeni fungi, i.e. Colletotrihum truntum, Rhizotoni solni, Slerotium rolfsii (Vughn nd Grdner 1993), Peniillium expnsum (Neri et l. 26), Alternri lternt (Hmilton-Kemp et l. 1992), Botrytis inere (And- -Nkpwtt et l. 26, Hmilton-Kemp et l. 1992, Kishimoto et l. 26). In this study the effet ws exmine of trns-2-hexenl nd trns-2-nonenl on the myelium growth of Phom nrissi (Aderh.) Boerem, de Gruyter et Noordel., the pthogen of Hippestrum nd other speies of fmily Amryllidee (Sniewsk 1998). * The reserh ws supported y Ministry of Siene nd Informtion Soiety Tehnologies, Grnt No 2 P6R 125 26. Roz. AR Pozn. CCCLXXXIII, Ogrodn. 41: 189-193 Wydwnitwo Akdemii Rolnizej im. August Cieszkowskiego w Poznniu, Poznń 27 PL ISSN 137-1738
19 A. Sniewsk, M. Sniewski Mteril nd methods The stok ulture of Phom nrissi ws mintined on potto-dextrose-gr (PDA- Merk) slnts t 25 C in the drk. Trns-2-hexenl nd trns-2-nonenl were purhsed from Sigm-Aldrih Chemils. These two ldehydes t finl onentrtion 1, 5, nd 25 μg m -3 in medium were used. These ompounds were dded to PDA fter steriliztion t temperture of out 5 C. Five mm dimeter plugs were tken from 7- dy-old ulture of P. nrissi, nd pled in the middle of 9 mm Petri dishes ontining PDA medium, supplemented erlier with tested ompounds. In nother experiment myelil plugs (5 mm dimeter) of P. nrissi tken from 7- -dy-old ulture were put in the enter of 9 mm Petri dishes with PDA nd on the ottom of other Petri dishes trns-2-hexenl or trns-2-nonenl were pled t dosge.1,.3 nd.5 μl per plte. Then pltes were seled together with prfilm (plte with P. nrissi ws over the dish with these ldehydes). Control pltes ontined the ultures growing on pure PDA, without ny dditions. Five Petri dishes were used s n experimentl unit nd the tril ws repeted twie. Inution ws onduted in drkness t 25 C. After two, four nd six dys of inution the dimeter of fungl olonies ws mesured in two perpendiulr diretions. The dt were sujeted to n nlysis of vrine nd the Dunn s t-test ws used to estimte the differenes etween mens t 5% level of signifine. Results nd disussion Trns-2-hexenl nd trns-2-nonenl showed strong inhiitory effet on the myelium growth of Phom nrissi when pplied diretly to PDA medium or pplied s voltiles. After 6-dy inution with oth of these ldehydes pplied lone to the medium t onentrtion of 25 μg m -3 the myelium growth of the pthogen ws totlly inhiited (Fig. 1, 2). Lower onentrtions of trns-2-nonenl proportionlly in lesser degree inhiited myelium growth; t onentrtion of 1 nd 5 μg m -3 in 22 nd 49%, respetively. In se of trns-2-hexenl the onentrtion of 1, 5 nd μg m -3 pplied to PDA medium to similr degree, y out 2%, inhiited myelium growth of P. nrissi. Trns-2-hexenl nd trns-2-nonenl pplied s voltiles t ll used onentrtions,.1,.3 nd.5 μl per 9 mm Petri dishes (i.e. 2; 6 nd 1 μg m -3 of free spe) gretly inhiited the myelium growth of P. nrissi; t onentrtion of 1 μg m -3 totlly inhiited myelium growth of the pthogen fter six dys inution (Fig. 3, 4). At onentrtion of 2 μg m -3 trns-2-hexenl limited the myelium growth of the pthogen in 25% ut trns-2-nonenl in 84% fter six-dy inution. Reently, Prost et l. (25) evluted ntimiroil tivities of 43 oxylipins, whih re lrge fmily of metolites derived from polyunsturted ftty ids, ginst set of 13 plnt pthogeni miroorgnisms inluding teri, oomyetes nd fungi, nd showed tht mny of the tested oxylipins re involved in defense ginst pthogens. Presumed funtions of oxylipins inlude diret ntimiroil effet, stimultion of plnt defense gene expression, nd regultion of plnt ell deth, ut the preise ontriution of individul oxylipins to plnt defense remins essentilly unknown (Prost et l. 25).
The effet of trns-2-hexenl nd trns-2-nonenl... 191 Inhiition of myelium growth (%) Hmownie wzrostu grzyni (%) 2 Dys of inution Dni inkuji Fig. 1. The effet of trns-2-hexenl pplied diretly to PDA medium on the myelium growth of Phom nrissi; vlues followed y the sme letter do not differ t 5% level of signifine (Dunn s test). Surfe of myelium growth in ontrol on PDA medium fter 2, 4 nd 6 dys of inution is 7.1; 2.9 nd 42.1 m 2, respetively Ry. 1. Wpływ trns-2-heksenlu dodnego ezpośrednio do pożywki PDA n wzrost grzyni Phom nrissi; wrtośi oznzone tą smą literą nie różnią się istotnie (5%) według testu Dunn. Powierzhni wzrostu grzyni w kontroli n pożywe PDA po 2, 4 i 6 dnih inkuji wynosi, odpowiednio 7,1; 2,9 i 42,1 m 2 1 μg m -3 5 μg m -3 μg m -3 25 μg m -3 Inhiition of myelium growth (%) Hmownie wzrostu grzyni (%) 2 d d 1 μg m -3 5 μg m -3 μg m -3 25 μg m -3 Dys of inution Dni inkuji Fig. 2. The effet of trns-2-nonenl pplied diretly to PDA medium on the myelium growth of Phom nrissi; vlues followed y the sme letter do not differ t 5% level of signifine (Dunn s test). Surfe of myelium growth in ontrol on PDA medium fter 2, 4 nd 6 dys of inution is 8.3; 25.7 nd 51.5 m 2, respetively Ry. 2. Wpływ trns-2-nonenlu dodnego ezpośrednio do pożywki PDA n wzrost grzyni Phom nrissi; wrtośi oznzone tą smą literą nie różnią się istotnie (5%) według testu Dunn. Powierzhni wzrostu grzyni w kontroli n pożywe PDA po 2, 4 i 6 dnih inkuji wynosi, odpowiednio 8,3; 25,7 i 51,5 m 2
192 A. Sniewsk, M. Sniewski Inhiition of myelium growth (%) Hmownie wzrostu grzyni (%) 2 2 μg m -3 6 μg m -3 1 μg m -3 Dys of inution Dni inkuji Fig. 3. The effet of trns-2-hexenl pplied s voltiles on the myelium growth of Phom nrissi; vlues followed y the sme letter do not differ t 5% level of signifine (Dunn s test). Surfe of myelium growth in ontrol on PDA medium fter 2, 4 nd 6 dys of inution is 7.2; 25.3 nd 45.3 m 2, respetively Ry. 3. Wpływ trns-2-heksenlu podnego w posti pr n wzrost grzyni Phom nrissi; wrtośi oznzone tą smą literą nie różnią się istotnie (5%) według testu Dunn. Powierzhni wzrostu grzyni w kontroli n pożywe PDA po 2, 4 i 6 dnih inkuji wynosi, odpowiednio 7,2; 25,3 i 45,3 m 2 Inhiition of myelium growth (%) Hmownie wzrostu grzyni (%) 2 2 μg m -3 6 μg m -3 1 μg m -3 Dys of inution Dni inkuji Fig. 4. The effet of trns-2-nonenl pplied s voltiles on the myelium growth of Phom nrissi; vlues followed y the sme letter do not differ t 5% level of signifine (Dunn s test). Surfe of myelium growth in ontrol on PDA medium fter 2; 4 nd 6 dys of inution is 7.2; 25.3 nd 45.8 m 2, respetively Ry. 4. Wpływ trns-2-nonenlu podnego w posti pr n wzrost grzyni Phom nrissi; wrtośi oznzone tą smą literą nie różnią się istotnie (5%) według testu Dunn. Powierzhni wzrostu grzyni w kontroli n pożywe PDA po 2; 4 i 6 dnih inkuji wynosi, odpowiednio 7,2; 25,3 i 45,8 m 2
The effet of trns-2-hexenl nd trns-2-nonenl... 193 Kishimoto et l. (26) showed tht (E)-2-hexenl nd (Z)-3-hexenl, enhne resistne ginst nerotrophi pthogen, Botrytis inere in Aridopsis thlin due to indution of lignifition, whih formed physil rrier to the penetrtion of the hyphe, nd the umultion of ntifungl sustnes inluding PDF1.2, PR-3 nd mlexin, whih inhiited the growth of the hyphe in the plnt ell. Conlusions 1. Trns-2-hexenl nd trns-2-nonenl, produed from polyunsturted ftty ids vi the lipoxygense pthwy, used gret inhiitory effet on the myelium growth of Phom nrissi, when pplied to PDA medium or pplied s voltiles. 2. It is prole tht trns-2-hexenl nd trns-2-nonenl ply n importnt role in disese resistne fter ttk y pthogens. Referenes And-Nkpwtt D., Krimm U., Shreier L., Shw W. (26): Dul ntgonism of ldehydes nd epiphyti teri from strwerry lef surfe ginst the pthogeni fungus Botrytis inere in vitro. BioControl 51: 279-291. Hmilton-Kemp T.R., MCrken C.T., Jr., Loughrin J.H., Andersen R.A., Hildernd D.F. (1992): Effets of some nturl voltile ompounds on the pthogeni fungi Alternri lternt nd Botrytis inere. J. Chem. Eol. 18: 183-191. Kishimoto K., Mtsui K., Ozw R., Tkyshi J. (26): Components of C6-ldehydeindued resistne in Aridopsis thlin ginst nerotrophi fungl pthogen, Botrytis inere. Plnt Si. 17: 715-723. Neri F., Mri M., Menniti A.M., Brigti S., Bertolini P. (26): Control of Peniillium expnsum in pers nd pples y trns-2-hexenl vpours. Posthrvest Biol. Tehnol. 41: 11-18. Prost I., Dhondt S., Rothe G., Viente J., Rodriguez M.J., Kift N., Cronne F., Griffiths G., Esquerre-Tugye M.-T., Roshl S., Cstresn C., Hmerg M., Fournier J. (25): Evlution of the ntimiroil tivities of plnt oxylipins supports their involvement in defense ginst pthogens. Plnt Physiol. 139: 192-1913. Sniewsk A. (1998): Czynniki iotyzne i iotyzne hmująe wzrost i rozwój Phom nrissi (Aderh.) Boerem, de Gruyter et Noordel., om. nov. Zesz. Nuk. Inst. Sdown. Kwi. Monogr. Rozpr.: 1-32. Vughn S.F., Grdner H.W. (1993): Lipoxygense-derived ldehydes inhiit fungi pthogeni on soyen. J. Chem. Eol. 19: 2337-2345. WPŁYW TRANS-2-HEKSENALU I TRANS-2-NONENALU NA WZROST GRZYBNI PHOMA NARCISSI IN VITRO Streszzenie Bdno wpływ trns-2-heksenlu i trns-2-nonenlu n wzrost grzyni Phom nrissi n pożywe PDA. Oydw ldehydy silnie hmowły wzrost grzyni P. nrissi, kiedy dodwno je do pożywki lu podwno w posti lotnej; większe inhiitorowe dziłnie wywierło zstosownie tyh ldehydów w posti lotnej. Hmująy wpływ trns-2-nonenlu n wzrost grzyni P. nrissi ył większy niż trns-2-heksenlu.