Plant growth regulatory activity in the phytopathogenic fungus Plectosphaerella melonis strain 502
Authors: Tsekhmister H. , Kyslynska A. , Kopilov E. , Nadkernychna O. 
Published: February 2022

Aim. To investigate the ability of our phytopathogenic fungal strain 502, earlier preliminarily identified as the phytopathogen
Plectosphaerella melonis (syn. Acremonium cucurbitacearum), to have phytotoxic and/or plant growth regulatory activity.
Methods. The phytotoxicity of strain 502, was studied by bioassays using the test cultures of corn (Zea mays L.), garden
cress (Lepidium sativum L.), cucumber (Cucumis sativus L.), and onion (Allium cepa L.). The cytotoxicity and genotoxicity
of the fungus were estimated using the Allium cepa-test. The mitotic index of the, the duration of mitosis phases, and the
frequency of aberrant ana-telophases of Allium cepa L. roots meristem was also investigated. For this purpose, strain 502,
was grown in the following culture media: synthetic Raulin-Thom medium for 10 days at 26 ± 2 °С. Cell-free filtrate
(culture fluid) was used for the study. Ethylene production was quantified in culture filtrate using gas-chromatography meth-
od. Ethylene measurement was performed every 7 days during 8 weeks. The determination was carried out using a gas
chromatograph «Agilent Technologies 6850» (USA) fitted with a flame ionization detector, using commercial ethylene
as a standard for identification and quantification Every experiment had three repeats. The reliability of experimental data
was assessed by statistical methods using Statistica 12 (Stat-Soft Inc., USA). Results. Undiluted culture fluid (obtained
by growing the fungus on liquid wort) of our strain 502 inhibited the growth of Z. mays seedlings by 14 %, L. sativum
seedlings by 18 % (1 : 100 dilution) and stimulated the growth of L. sativum roots by 54 and 41 % (1 : 10 and 1 : 100
dilutions, respectively). The culture fluid, obtained by growing the fungus on Raulin-Thom’s synthetic agar, demonstrated
a slight inhibitory effect on the seedlings and roots of L. sativum, and at the dilution of 1 : 1000 stimulated growth by
30 %. Insignificant changes in the mitotic index of the meristem of A. cepa roots were revealed at the effect of the culture
fluid of P. melonis, strain 502, diluted at the ratio of 1 : 100 and 1 : 1000. At the same time, the number of cells at the prophase
stage decreased 1.7 times (1 : 100 dilution). There is a significant increase in the number of cells at the metaphase stage –
1.3 and 1.4 times (dilution 1 : 100 and 1 : 1000, respectively), the anaphase stage – 2.1 and 1.8 times (dilution 1 : 100 and
1 : 1000, respectively) and the telophase stage – 1.8 times (1 : 100 dilution), as compared with the positive control
(culture medium). The frequencies of aberrant ana-telophases in the apical meristems of the initial roots were
5.0 and 2.2 % (at the culture fluid dilution of 1 : 100 and 1 : 1000, respectively). We researched the abil-
ity of P. melonis 502 to synthesize ethylene and the highest level of it was registered after 5 weeks of cultivation
(111.78 nmol/h g). Conclusions: It was demonstrated by us that the culture fluid of strain 502 showed no phytotoxic effect
on roots and seedlings of the investigated cultures, demonstrating the exclusion of phytotoxins from the possible range of
effectors. No cytotoxic or genotoxic activity of the culture fluid was observed either. However, the culture fluid altered the
dynamics of the cell cycle, in particular, shortened the prophase and stimulated the metaphase, anaphase, and telophase. The
culture fluid of the fungus stimulated the growth of L. sativum roots depending on the nutrient medium, where the fungus
was grown and cultivated. In particular, when growing the fungus on the liquid wort, the growth was higher by 54 and 41 %
(dilution 1 : 10 and 1 : 100, respectively), when growing on synthetic Raulin-Thom’s medium – by 30 %. This demonstrates
the ability of strain 502 to possibly synthesize growth promoting substances. Also, we have shown the ability of this strain to
synthetize ethylene in vitro (111.78 ± 13.27 nmol/h per g), which can act as virulence factor. We consider the obtained results
to be the first stage of the study on the mechanism of the interaction between pathogenic strain 502 and plants.