Rocz. AR Pozn. CCCLXXXIII, Ogrodn. 41: 189-193
© Wydawnictwo Akademii Rolniczej im. Augusta Cieszkowskiego w Poznaniu, Poznań 2007 PL ISSN 0137-1738
ALICJA SANIEWSKA, MARIAN SANIEWSKI
THE EFFECT OF TRANS-2-HEXENAL AND TRANS-2-NONENAL ON THE MYCELIUM GROWTH OF PHOMA NARCISSI
IN VITRO
*From Research Institute of Pomology and Floriculture in Skierniewice
ABSTRACT. The effect of trans-2-hexenal and trans-2-nonenal on the mycelium growth of Phoma narcissi on PDA medium was investigated. Both aldehydes strongly inhibited the mycelium growth of P. narcissi when applied to PDA medium or applied as volatiles; greater inhibitory effect was observed when they were applied as volatiles. The inhibitory effect on the mycelium growth of P. narcissi was greater of trans-2-nonenal than trans-2-hexenal.
Key words: trans-2-hexenal, trans-2-nonenal, Phoma narcissi, mycelium growth
Introduction
Several unsaturated aldehydes are produced from polyunsaturated fatty acids via the lipoxygenase pathway when plants are wounded mechanically or infected by pathogens.
Several of these aldehydes, including trans-2-hexenal and trans-2-nonenal are found to be inhibitory to pathogenic fungi, i.e. Colletotrichum truncatum, Rhizoctonia solani, Sclerotium rolfsii (Vaughn and Gardner 1993), Penicillium expansum (Neri et al.
2006), Alternaria alternata (Hamilton-Kemp et al. 1992), Botrytis cinerea (Abanda- -Nkpwatt et al. 2006, Hamilton-Kemp et al. 1992, Kishimoto et al. 2006).
In this study the effect was examine of trans-2-hexenal and trans-2-nonenal on the mycelium growth of Phoma narcissi (Aderh.) Boerema, de Gruyter et Noordel., the pathogen of Hippeastrum and other species of family Amaryllidaceae (Saniewska 1998).
*The research was supported by Ministry of Science and Information Society Technologies, Grant No 2 P06R 125 26.
Material and methods
The stock culture of Phoma narcissi was maintained on potato-dextrose-agar (PDA- Merck) slants at 25°C in the dark. Trans-2-hexenal and trans-2-nonenal were purchased from Sigma-Aldrich Chemicals. These two aldehydes at final concentration 10, 50, 100 and 250 μg·cm-3 in medium were used. These compounds were added to PDA after sterilization at temperature of about 50°C. Five mm diameter plugs were taken from 7- day-old culture of P. narcissi, and placed in the middle of 90 mm Petri dishes contain- ing PDA medium, supplemented earlier with tested compounds.
In another experiment mycelial plugs (5 mm diameter) of P. narcissi taken from 7- -day-old culture were put in the center of 90 mm Petri dishes with PDA and on the bot- tom of other Petri dishes trans-2-hexenal or trans-2-nonenal were placed at a dosage 0.1, 0.3 and 0.5 μl per plate. Then plates were sealed together with parafilm (plate with P. narcissi was over the dish with these aldehydes). Control plates contained the cul- tures growing on pure PDA, without any additions. Five Petri dishes were used as an experimental unit and the trial was repeated twice. Incubation was conducted in dark- ness at 25°C. After two, four and six days of incubation the diameter of fungal colonies was measured in two perpendicular directions.
The data were subjected to an analysis of variance and the Duncan’s t-test was used to estimate the differences between means at 5% level of significance.
Results and discussion
Trans-2-hexenal and trans-2-nonenal showed strong inhibitory effect on the myce- lium growth of Phoma narcissi when applied directly to PDA medium or applied as volatiles. After 6-day incubation with both of these aldehydes applied alone to the me- dium at a concentration of 250 μg·cm-3 the mycelium growth of the pathogen was totally inhibited (Fig. 1, 2). Lower concentrations of trans-2-nonenal proportionally in a lesser degree inhibited mycelium growth; at a concentration of 10 and 50 μg·cm-3 in 22 and 49%, respectively. In case of trans-2-hexenal the concentration of 10, 50 and 100 μg·cm-3 applied to PDA medium to a similar degree, by about 20%, inhibited mycelium growth of P. narcissi.
Trans-2-hexenal and trans-2-nonenal applied as volatiles at all used concentrations, 0.1, 0.3 and 0.5 μl per 90 mm Petri dishes (i.e. 2; 6 and 10 μg·cm-3 of free space) greatly inhibited the mycelium growth of P. narcissi; at a concentration of 10 μg·cm-3 totally inhibited mycelium growth of the pathogen after six days incubation (Fig. 3, 4). At a concentration of 2 μg·cm-3 trans-2-hexenal limited the mycelium growth of the patho- gen in 25% but trans-2-nonenal in 84% after six-day incubation.
Recently, Prost et al. (2005) evaluated antimicrobial activities of 43 oxylipins, which are a large family of metabolites derived from polyunsaturated fatty acids, against a set of 13 plant pathogenic microorganisms including bacteria, oomycetes and fungi, and showed that many of the tested oxylipins are involved in defense against pathogens. Presumed functions of oxylipins include direct antimicrobial effect, stimula- tion of plant defense gene expression, and regulation of plant cell death, but the precise contribution of individual oxylipins to plant defense remains essentially unknown (Prost et al. 2005).
Fig. 1. The effect of trans-2-hexenal applied directly to PDA medium on the mycelium growth of Phoma narcissi; values followed by the same letter do not differ at 5% level of significance (Duncan’s test). Surface of mycelium growth in control on PDA medium after
2, 4 and 6 days of incubation is 7.1; 20.9 and 42.1 cm2, respectively
Ryc. 1. Wpływ trans-2-heksenalu dodanego bezpośrednio do pożywki PDA na wzrost grzybni Phoma narcissi; wartości oznaczone tą samą literą nie różnią się istotnie (5%) we- dług testu Duncana. Powierzchnia wzrostu grzybni w kontroli na pożywce PDA po 2, 4 i 6
dniach inkubacji wynosi, odpowiednio 7,1; 20,9 i 42,1 cm2
Fig. 2. The effect of trans-2-nonenal applied directly to PDA medium on the mycelium growth of Phoma narcissi; values followed by the same letter do not differ at 5% level of significance (Duncan’s test). Surface of mycelium growth in control on PDA medium after 2, 4 and 6 days of incubation is 8.3; 25.7 and 51.5 cm2, respectively
Ryc. 2. Wpływ trans-2-nonenalu dodanego bezpośrednio do pożywki PDA na wzrost grzyb- ni Phoma narcissi; wartości oznaczone tą samą literą nie różnią się istotnie (5%) według te- stu Duncana. Powierzchnia wzrostu grzybni w kontroli na pożywce PDA po 2, 4 i 6 dniach
inkubacji wynosi, odpowiednio 8,3; 25,7 i 51,5 cm2 d d
c
c c
b b
b a
a a
a
0 20 40 60 80 100 120
2 4 6
Days of incubation – Dni inkubacji Inhibition of mycelium growth (%) Hamowanie wzrostu grzybni (%)
10 μg·cm-3 50 μg·cm-3 100 μg·cm-3 250 μg·cm-3 b b
c
b b c
b b b
a a
a
0 20 40 60 80 100 120
2 4 6
Days of incubation – Dni inkubacji Inhibition of mycelium growth (%) Hamowanie wzrostu grzybni (%)
10 μg·cm-3 50 μg·cm-3 100 μg·cm-3 250 μg·cm-3
Fig. 3. The effect of trans-2-hexenal applied as a volatiles on the mycelium growth of Phoma narcissi; values followed by the same letter do not differ at 5% level of significance (Duncan’s test). Surface of mycelium growth in control on PDA medium after 2, 4 and 6
days of incubation is 7.2; 25.3 and 45.3 cm2, respectively
Ryc. 3. Wpływ trans-2-heksenalu podanego w postaci par na wzrost grzybni Phoma narcis- si; wartości oznaczone tą samą literą nie różnią się istotnie (5%) według testu Duncana. Po- wierzchnia wzrostu grzybni w kontroli na pożywce PDA po 2, 4 i 6 dniach inkubacji wynosi,
odpowiednio 7,2; 25,3 i 45,3 cm2
Fig. 4. The effect of trans-2-nonenal applied as a volatiles on the mycelium growth of Phoma narcissi; values followed by the same letter do not differ at 5% level of significance (Duncan’s test). Surface of mycelium growth in control on PDA medium after 2; 4 and 6
days of incubation is 7.2; 25.3 and 45.8 cm2, respectively
Ryc. 4. Wpływ trans-2-nonenalu podanego w postaci par na wzrost grzybni Phoma narcissi;
wartości oznaczone tą samą literą nie różnią się istotnie (5%) według testu Duncana. Po- wierzchnia wzrostu grzybni w kontroli na pożywce PDA po 2; 4 i 6 dniach inkubacji wynosi,
odpowiednio 7,2; 25,3 i 45,8 cm2 a
a a a a
a a
a a
0 20 40 60 80 100 120
2 4 6
Inhibition of mycelium growth (%) Hamowanie wzrostu grzybni (%)
2 μg·cm-3 6 μg·cm-3 10 μg·cm-3
Days of incubation – Dni inkubacji c c
c b b
b
a a
a
0 20 40 60 80 100 120
2 4 6
Inhibition of mycelium growth (%) Hamowanie wzrostu grzybni (%)
2 μg·cm-3 6 μg·cm-3 10 μg·cm-3
Days of incubation – Dni inkubacji
Kishimoto et al. (2006) showed that (E)-2-hexenal and (Z)-3-hexenal, enhance re- sistance against a necrotrophic pathogen, Botrytis cinerea in Arabidopsis thaliana due to induction of lignification, which formed a physical barrier to the penetration of the hyphae, and the accumulation of antifungal substances including PDF1.2, PR-3 and camalexin, which inhibited the growth of the hyphae in the plant cell.
Conclusions
1. Trans-2-hexenal and trans-2-nonenal, produced from polyunsaturated fatty acids via the lipoxygenase pathway, caused great inhibitory effect on the mycelium growth of Phoma narcissi, when applied to PDA medium or applied as volatiles.
2. It is probable that trans-2-hexenal and trans-2-nonenal play an important role in disease resistance after attack by pathogens.
References
Abanda-Nkpwatt D., Krimm U., Schreiber L., Schwab W. (2006): Dual antagonism of alde- hydes and epiphytic bacteria from strawberry leaf surface against the pathogenic fungus Bo- trytis cinerea in vitro. BioControl 51: 279-291.
Hamilton-Kemp T.R., McCracken C.T., Jr., Loughrin J.H., Andersen R.A., Hildebrand D.F. (1992): Effects of some natural volatile compounds on the pathogenic fungi Alternaria alternata and Botrytis cinerea. J. Chem. Ecol. 18: 1083-1091.
Kishimoto K., Matsui K., Ozawa R., Takabayashi J. (2006): Components of C6-aldehyde- induced resistance in Arabidopsis thaliana against a necrotrophic fungal pathogen, Botrytis cinerea. Plant Sci. 170: 715-723.
Neri F., Mari M., Menniti A.M., Brigati S., Bertolini P. (2006): Control of Penicillium expan- sum in pears and apples by trans-2-hexenal vapours. Postharvest Biol. Technol. 41: 101-108.
Prost I., Dhondt S., Rothe G., Vicente J., Rodriguez M.J., Kift N., Carbonne F., Griffiths G., Esquerre-Tugaye M.-T., Roshal S., Castresana C., Hamberg M., Fournier J. (2005):
Evaluation of the antimicrobial activities of plant oxylipins supports their involvement in de- fense against pathogens. Plant Physiol. 139: 1902-1913.
Saniewska A. (1998): Czynniki biotyczne i abiotyczne hamujące wzrost i rozwój Phoma narcissi (Aderh.) Boerema, de Gruyter et Noordel., comb. nov. Zesz. Nauk. Inst. Sadown. Kwiac.
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Vaughn S.F., Gardner H.W. (1993): Lipoxygenase-derived aldehydes inhibit fungi pathogenic on soybean. J. Chem. Ecol. 19: 2337-2345.
WPŁYW TRANS-2-HEKSENALU I TRANS-2-NONENALU NA WZROST GRZYBNI PHOMA NARCISSI IN VITRO
S t r e s z c z e n i e
Badano wpływ trans-2-heksenalu i trans-2-nonenalu na wzrost grzybni Phoma narcissi na pożywce PDA. Obydwa aldehydy silnie hamowały wzrost grzybni P. narcissi, kiedy dodawano je do pożywki lub podawano w postaci lotnej; większe inhibitorowe działanie wywierało zastoso- wanie tych aldehydów w postaci lotnej. Hamujący wpływ trans-2-nonenalu na wzrost grzybni P. narcissi był większy niż trans-2-heksenalu.
