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Brief Report

Psychrotolerant Strains of Phoma herbarum with Herbicidal Activity

by
Andrey Trigubovich
1,
Maryna Mandryk-Litvinkovich
2,
Anastasiya Valakhanovich
1,
Elizaveta Gorodilova
3,
Daniil Malygin
4,5,
Emiliya Kalamiyets
2 and
Sofia Sokornova
4,5,*
1
Institute of Microbiology of the National Academy of Sciences of Belarus, 2 Kuprevich Str.,220084 Minsk, Belarus
2
State Scientific Production Association “Chemical Synthesis and Biotechnology”, 2 Kuprevich Str.,220141 Minsk, Belarus
3
Research Resource Center for Molecular and Cell Technologies, Saint-Petersburg State University, 17 Botanicheskaya Str., 198504 Saint-Petersburg, Russia
4
All-Russian Institute of Plant Protection, 3 Podbelsokogo Rd., 196608 Saint-Petersburg, Russia
5
Center for Algorithmic Biotechnology, St. Petersburg State University, 7/9 Universitetskaya Emb.,199034 Saint-Petersburg, Russia
*
Author to whom correspondence should be addressed.
Agronomy 2023, 13(6), 1619; https://doi.org/10.3390/agronomy13061619
Submission received: 25 May 2023 / Revised: 9 June 2023 / Accepted: 13 June 2023 / Published: 15 June 2023
(This article belongs to the Special Issue Pests, Pesticides and Food Safety in a Changing Climate)
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Abstract

The search for stress-tolerant producer strains is a key factor in the development of biological mycoherbicides. The aim of the study was to assess the herbicidal potential of phoma-like fungi. Morphological and physiological features of two Antarctic psychrotolerant strains 20-A7-1.M19 and 20-A7-1.M29 were studied. Multilocus sequence analysis was used to identify these strains. They happened to belong to Phoma herbarum Westend. The psychrotolerant properties of these strains were suggested not only by ecology, but also by their capability to grow in a wide temperature range from 5 °C to 35 °C, being resistant to high insolation, UV radiation, aridity, and other extreme conditions. It was shown that treatment with their cell-free cultural fugate, crude mycelium extract, and culture liquid significantly reduced the seed germination of troublesome weeds such as dandelion and goldenrod. Cell-free cultural fugate and culture liquid also led to the formation of chlorosis and necrotic spots on leaves. Thus, psychrotolerant strains P. herbarum 20-A7-1.M19 and 20-A7-1.M29 demonstrate high biotechnological potential. Our next step is to determine the structures of biologically active substances and to increase their biosynthesis, as well as the development of biological and biorational mycoherbicides. New mycoherbicides can reduce the chemical load on agroecosystems and increase the effectiveness of applied chemicals.

1. Introduction

Reducing the chemical load on agroecosystems is one of the global goals in the development of breakthrough agrobiotechnologies that can be achieved, including via the application of bio- and biorational mycoherbicides [1]. However, unstable and insufficiently high effectiveness of biological plant protection products in field conditions is a common cause of a limitation of their wide spreading. Microorganisms that live in extreme climate habitats are promising research directions because they show high viability and resistance to adverse abiotic factors [2]. At the same time, most of these studies have been carried out mainly on Antarctic bacterial strains, while almost no attention has been paid to the activity of micromycetes of Antarctic substrates [3,4].
The most studied polar producer strains of micromycetes belong to the genus Penicillium [5]. Antarctica species of other genera, such as Phoma, also show the ability to produce metabolites that are promising for agrobiotechnology [6,7]. Phoma-like species are a diverse group of Ascomycetes fungi that are widespread in nature and occupy various ecological niches in different regions of the world. Their ability to colonize different substrates is explained by the production of biologically active substances that provide protective functions, such as various pigments, enzymes, polysaccharides, etc. [8]. They are also known for their ability to produce specialized metabolites such as phomanolide, phomodione, cercosporamide, cytochalasins, macrocidin, herbarumin, anthraquinone pigments, etc. These metabolites demonstrate insecticide, antiviral, fungistatic, antibacterial, and herbicide activities [9,10,11].
The development of mycopesticides should include the accurate identification of species, which allows for a more complete characterization of producer strains. Correct identification makes it possible to reveal a correlation between the phylogenetic relationships of fungi and their metabolomics and conduct molecular barcoding. The identification of micromycetes is most often limited to traditional phenotype-based identification methods and the molecular analysis of the internal transcribed spacers (ITSs) region [12,13]. It should be noted that morphological characteristics of phoma-like fungi are unstable and can overlap not only in different species, but also in genera [14,15]. As a result, inaccurate identification does not allow for predicting useful biotechnological characteristics of producer strains and their environmental safety. Thus, the correct identification of producer strains using multilocus analysis allows for a qualitative selection of potential mycoherbicides. Molecular barcoding is required for producer strain certification. To assess environmental risks, it is important to predict their ability to produce mycotoxin.
This area of research is especially relevant for novel strains of Phoma that are found in the Antarctic and have adapted to a harsh climate and extreme conditions: low temperatures in the short summer period and high insolation, UV radiation, and aridity. Such extreme impacts can lead to the formation of new genotype variants, which can be interesting for molecular genetics research, establishing their phylogenetic position and connection with the production of active substances [7].
The aim was to assess the biotechnological potential of psychrotolerant phoma-like fungi. The objectives of the study included clarifying the identification of strains using molecular and classical mycological methods, evaluating the fungus growth rate, and estimating herbicidal activity against troublesome Asteraceae species: dandelion (Taraxacum officinale) and goldenrod (Solidago canadensis).

2. Materials and Methods

Object of study. Two Phoma spp. strains 20-A7-1.M19 and 20-A7-1.M29 were isolated from mold lesions. These lesions were found on samples of anthropogenic materials delivered from the construction site of the Belarusian Antarctic Station, collected during the 7th Belarusian Antarctic Expedition (2014–2015) in extreme conditions of East Antarctica, Enderby Land, oasis Mount Vechernyaya. The strains 20-A7-1.M19 and 20-A7-1.M29 were deposited in the Belarusian collection of non-pathogenic microorganisms as BIM F-795 G and BIM F-796 G, respectively. They were stored on Czapek-Dox agar medium slants in test tubes at 4 °C.
Deep liquid fermentation. For the initial culture production, small pieces of mycelium were aseptically transferred to Czapek-Dox agar plates and incubated for 14 days at 24 °C in the dark. Mycelial plugs (5 mm in diameter) from the colony margins were used for the inoculation of liquid media. The Czapek-Dox broth medium was used to obtain the cultural liquid and mycelial biomass by incubating it at 25 °C and at 200 rpm for 5–7 days (stationary growth phase).
Traditional phenotype-based identification. The study of cultural and morphological features was carried out according to the protocol of Boerema G.H. using the following nutrient media: plain agar, malt agar, potato-dextrose agar, and Czapek-Dox agar medium with various carbon sources [16]. Microphotographs of pycnidia and conidia of strains 20-A7-1.M19 and 20-A7-1.M29 were obtained using a Leica M165 Stereo Microscope equipped with a digital camera.
Multilocus phylogenetic analysis. The strains 20-A7-1.M19 and 20-A7-1.M29 were analyzed using multilocus analysis including the amplification and sequencing of large subunit ribosomal DNA (LSU), and partial beta-tubulin and elongation factor genes’ loci. The genomic DNA of the fungus was extracted using the standard acetyltrimethylammonium bromide (CTAB)/chloroform protocol with Sambrook modification using an Ultraclean Microbial DNA Isolation Kit (Mo Bio Laboratories, Carlsbad, CA, USA) [17]. For nucleotide sequence comparisons, the fragments of three loci were analyzed: ITS, partial tef1, and tub2. Amplification of ITS was conducted utilizing the following primer combinations: ITS1 (5′-TCCGTAGGTGAACCTGCGG-3′) and ITS4 (5′-TCCTCCGCTTATTGATATGC-3′); partial tef1: EF1-728F (5′-CTCGAGAAGTTCGAGAAGG-3′) and EF1-986R (5′-TACTTGAAGGAACCCTTACC-3′); and tub2: βtub2Fw (5′-GTBCACCTYCARACCGGYCARTG-3′) and βtub4Rd (5′-CCRGAYTGRCCRAARACRAAGTTGTC-3′) [18,19,20,21] (Figure S1). The PCR conditions were executed according to the protocol of the HotStarTaq Master Mix Kit (Quagene). The PCRs were performed in a Tercik Thermal Cycler (DNA-Technology, Moscow, Russia) in a total volume of 20 µL. Sanger sequencing was performed at the Research Park «Centre for Molecular and Cell Technologies Center» of Saint Petersburg State University. Consensus sequences were computed from the forward and reverse sequences using the MEGA v. 7.24 software package. The consensus sequences were deposited in GenBank.
Metabolite extraction. Submerged culture was used for obtaining crude mycelium extract (CME), containing pigments. The culture liquid (CL) was separated from the mycelium via centrifugation at 5000 rpm for 10 min. The biomass was washed with distilled water, then dried with running air at 24 °C and ground in a homogenizer. The extraction of mycelium was carried out with ethyl acetate (EtAc) in an ultrasonic bath for 30 min, three times. Then, the extracts were combined and evaporated. The yield of crude extracts was estimated by the weight.
Chromatographic and spectrometric analyses. Optical density was evaluated using a spectrophotometer, Shimadzu UV-2401PC UV-VIS.
GC MS/MS analysis was performed using an Agilent5860 chromatograph and Agilent ChemStation E.02.02.1431 software (Agilent Technologies, Santa Clara, CA, USA). Separation was conducted with a capillary column 30 m long, 0.25 mm in diameter, and stationary phase film (95% dimethylpolyoxane, 5% diphenyl), at a thickness of 0.1 μm. The following conditions were used: helium flow rate of 1 mL/min−1, and evaporator temperature of 230 °C, at a flow split ratio of 1:20. The temperature conditions of the column thermostat were the following: initial temperature of 70 °C, increased by 6 °C/min−1 up to 340 °C. The peaks were recorded using an Agilent 5975S mass selective detector (Agilent Technologies, Santa Clara, CA, USA) in the total ion recording mode with a frequency of 1.8 scans per second. Electron impact ionization was performed at 70 V under an ion source temperature of 230 °C.
HPLC MS/MS analysis was performed using a Shimadzu LCMS-IT-TOF-8030 (Shimadzu Corporation, Kyoto, Japan). The separation was performed on a C18 column (4.9 × 250 mm, 4 µm) (Water Milford, MA, USA). The mobile phase consisted of 1% aqueous formic acid (solvent A) and 1% formic acid in acetonitrile/methanol (25/75) (solvent B). Separation was achieved using the following elution gradient: 2% B isocratic for 10 min, from 2% to 98% B linear for 30 min, and 98% B isocratic for 7 min. The flow rate was 0.6 mL/min, and the injection volume was 10 µL. The column oven was set at 30 °C. The mass spectral data were acquired using the following ESI inlet conditions: nebulizing gas and drying gas were nitrogen at a flow rate of 3.0 and 15.0 L/min, respectively; the interface voltage was set to −3.5 kV; the desolvation line temperature was 250 °C; and the heat block temperature was 400 °C. The mass spectrometer was operated in the negative ion scan and product ion scan mode using analyte-specific precursor ions, with argon as the CID (collision-induced dissociation) gas at a pressure of 230 kPa.
Seed germination bioassay. Seeds of T. officinale and S. canadensis were first dipped in 70% ethyl alcohol (C2H5OH) for 30 s and in 5% sodium hypochlorite (NaOCl) for 30 min [22]. After that, they were dipped in 10% CL, cell-free cultural fugate (CFCF), and CME for 20 min. Then, they were placed on wet filter paper in Petri dishes and incubated at room temperature. The seed germination percent was recorded after 7 days. Seeds incubated in sterilized distilled water served as the control.
Detached leaf bioassay. Detached leaf bioassay was carried out according to Sokornova, Berestetskiy (2018) [23]. Surface-sterilized (70% C2H5OH) weed leaves were treated with 10% CL, CFCF, and CME, and were incubated at 28 ± 2 °C for 2 days. A positive reaction was indicated by the appearance of chlorotic and necrotic spots.
Data analysis. The bioassays were performed three times. Data were subjected to one-way and two-way ANOVA followed by the comparison of multiple treatment levels, with the control demonstrating the least significant difference (LSD) at p = 95%. The statistical analyses were performed using Statistica 8.0 (StatSoft, Tusla, OK, USA).

3. Results

Phoma-like fungi are known for their resistance to unfavorable environmental factors and for being widespread around the world. They can remain viable in soil and other substrates for a long time, which contributes to their distribution even in unfavorable conditions [24]. Phoma-like fungi are a large non-taxonomic group of asexual ascomycetes. Their system is complex due to high morphological variability, polymorphism, and the absence of specific reproductive organs in most species [7,16,20]. Therefore, for accurate species identification, it is necessary to use modern methods of molecular genetics in combination with morphological characteristics. The phoma-like strains that we studied have some similar characteristics: both can produce pigments and melanized pycnidia on Czapek-Dox agar. The color of these pigments varies from reddish-violet to yellowish-brown, and the color of melanized pycnidia varies from brown to black. At the same time, visible morphological differences do not make it possible to reliably determine whether these are different species or a manifestation of strain differences in fungi of the same species (Figure 1).
It should be noted that at the beginning of growth, the hyphae were light, and then darkened. The maximum pigmentation of the mycelium and the cultivation medium was observed on the 7th day of cultivation at a lower temperature of 15 °C. Within 10–20 days, numerous spherical dark pycnidia were formed. Their size ranged from 400 to 500 μm in diameter. The pycnidia contained single slightly oval pycnidiospores without septa, 5–7 μm × 2–3 μm in size. Both strains had significant cultural differences between colonies. However, the study of the microscopic structure of sporulation organs did not show significant differences, and their appearance was typical for Phoma herbarum.
Both strains can show psychrotolerant properties in a wide temperature range from 5 °C to 35 °C. The optimal growth temperature of the strains 20-A7-1.M19 and 20-A7-1.M29 is near 28 °C. However, increasing the temperature of cultivation to 37 °C has a certain fungistatic effect. The linear growth rate on the Czapek-Dox agar is the highest for the 20-A7-1.M29 strain at 28 °C and reaches 7.4 ± 0.3 mm/day. At the same temperature, the growth rate of the 20-A7-1.M19 strain was a bit lower—6.7 ± 0.3 mm/day. Lowering the cultivation temperature to 15 °C led to an increase in the lag phase time but did not significantly reduce the growth rate—5.7 ± 0.2 mm/day for both strains. At 5 °C, the development of colonies began on the 9th day. At this temperature, the 20-A7-1.M19 strain showed a growth rate twice that of the 20-A7-1.M29 strain—4.3 ± 0.2 mm/day and 1.6 ± 0.2 mm/day, respectively. Thus, the 20-A7-1.M19 strain is more resistant to low temperatures, but susceptible to high ones.
Lower temperatures favor the formation of the dormant stage structures of P. herbarum. So, at temperatures from 5 to 15 °C, pycnidia are formed on the 10–20th day. On minimal cultivation media without carbon source P. herbarum, the 20-A7-1.M19 and 20-A7-1.M29 strains form pycnidia up to 7 days. The addition of 2% glucose slows down this process.
Phylogenetic analysis based on sequences from three loci, including translation elongation factor 1-α (EF1-α), β-tubulin (TUB) genes, and the rDNA internal transcribed spacer (ITS), was performed (Figure S1). It indicated that Phoma strains 20-A7-1.M19 and 20-A7-1.M29 are clustered in a distinct clade closely related to Phoma herbarum (Figure 2; Table S1). Thus, multilocus sequence analysis identified these strains as belonging to the species Phoma herbarum.
There are data that indicate that phoma-like micromycetes produce many phytotoxic metabolites, such as macrocidin, herbarumin, phaeosphaeride, anthraquinone pigments, etc. [7,25]. So, we studied the herbicidal activity of CL, CFCF, and CME of P. herbarum 20-A7-1.M19 and P. herbarum 20-A7-1.M29 on the seeds and leaves of the common troublesome weeds dandelion (T. officinale) and goldenrod (S. canadensis). Preliminary experiments have shown that the herbicidal activity correlates with the absorption of the samples at 460 nm (Figure S2). Therefore, the maximum production of the desired substances was observed in the middle of the stationary growth phase of mycelium. The deep fermentation was stopped at OD460 ≥ 0.25. The spectrophotometry data show that there are at least two peaks at 460 and 480 nm.
The assessment of metabolite fingerprinting of CME was performed using high-performance liquid and gas chromatography with mass spectrometry. It showed similarities of metabolic profiles of the 20-A7-1.M19 and 20-A7-1.M29 strains (Figure S2).
The results of impact of CL, CFCF, and CME on seed germination are represented in Table 1. In the control samples treated with distilled water, the seed germination rate reached 70–80%. CL and CFCF suppressed seed germination. The addition of CL or CFCF of P. herbarum 20-A7-1.M19 suppressed the germination of 40–42% of dandelion seeds and 30–57% of goldenrod seeds. The suppression of dandelion seed germination by CL, CFCF, and CME P. herbarum 20-A7-1.M29 was 30, 40, and 60%, respectively. The suppression of goldenrod seed germination by CL, CFCF, and CME P. herbarum 20-A7-1.M29 was 55, 65, and 40%, respectively. These data indicate that CL, CFCF, and CME show a herbicidal effect on selected troublesome weeds.
Since the germination percentages only reflect radicle emergence from the seed coat, we decided to estimate the effect on plants in situ. Detached leaf bioassay was performed on leaves treated with CL, CFCF, and CME of P. herbarum 20-A7-1.M19 and P. herbarum 20-A7-1.M29.
The damage caused to T. officinale and S. canadensis by the P. herbarum 20-A7-1.M19 and P. herbarum 20-A7-1.M29 strains was similar. The treatment of leaves with CL and CFCF led to severe chlorosis, yellowing, wilting, and the appearance of necrotic spots, which were more pronounced on dandelion (Figure 3). The leaf damage ranged from 80 to 90%. With such damage, plants spend additional energy restoring themselves, and in most cases die. The experiments were carried out in vitro; therefore, to test herbicidal activity, it is necessary to conduct experiments in field conditions. It is important to note that these P. herbarum strains show herbicidal, not phytotoxic, activity. The main difference is that substances with phytotoxic activity require initial damage to the leaves to infect the plant, while substances with herbicidal activity can damage the leaves anyway.

4. Discussion

In this study, it was found that the psychrotolerant strains 20-A7-1.M19 and 20-A7-1.M29 isolated from anthropogenic wastes have some differences in their morphological characteristics but belong to the same P. herbarum species. The evaluation of the morphological and physiological features of two Antarctic psychrotolerant P. herbarum strains 20-A7-1.M19 and 20-A7-1.M29 showed that they grow rapidly and form pycnidia on poor substrates at low temperatures. This suggests that they will be stress-tolerant to adverse conditions in the field. We confirmed that the P. herbarum 20-A7-1.M19 and 20-A7-1.M29 strains show herbicidal potential. They inhibit seed germination and cause leaf spot of dandelion and goldenrod from the Asteraceae family.
Since the optical density peak is at 460 nm, we assume that our substance is a pigment.
The metabolite fingerprinting obtained via chromatography and spectrometry differed from known fungal extracts with herbicidal activities (Table 2; Figure S2). The metabolic profile data suggest that pigments show herbicidal activity. The main groups of pigments include simple benzoquinones, anthraquinones, naphthoquinones, terphenylquinones, pulvinic acids and the derived products, terpenoid quinines, benzotropolones, compounds of fatty acid origin, and nitrogen-containing substances (betalains and other alkaloids) [26]. Our next step is determining the structures of herbicidal active substances.
It is worth noting that mycoherbicides and biorational herbicides were successfully developed based on P. herbarum and their metabolites [31,32,33,34,35,36,37]. For example, a phytotoxic anthraquinone, which was identified as anhydropseudophlegmacin-9,10-quinone-3′-amino-8′-O-methyl ether, was isolated from a phytopathogenic fungus P. herbarum FGCC#54 strain [38]. This pigment is rarely found in nature. However, via the study of metabolites of the endophytic biota of an obnoxious weed Parthenium hysterophorus (Asteraceae family), anhydropseudophlegmacin-9,10-quinone-3′-amino-8′-O-methyl ether was detected. The assessment of the herbicidal potential of this pigment against common weeds of Central India was carried out, and a positive result was obtained [39]. Furthermore, it should be noted that phoma-like fungi can lead endophytic lifestyles [40]. Phytotoxine lactones herbarumin I–III were also isolated from P. herbarum [41,42]. Derivates with higher toxicity were also developed based on herbarumin I–III [32]. Another herbicidal active compound (phaeosphaeride A) is produced by Phoma sp. [25].
Both the P. herbarum 20-A7-1.M19 and 20-A7-1.M29 strains showed high herbicide activity on dandelion and goldenrod. Strain differences are manifested in the synthesis of specialized metabolites. Seed treatment with culture liquid, as well as cell-free cultural fugate and crude mycelium extract, significantly reduces the germination of these troublesome plants and can be applied for weed control.
Thus, the P. herbarum 20-A7-1.M19 and 20-A7-1.M29 strains can be considered as potential biological and biorational mycoherbicides.

5. Conclusions

The search for new potential mycoherbicides is very important for the agriculture safety purpose of weed control. We suppose that the Phoma herbarum 20-A7-1.M19 and 20-A7-1.M29 strains are promising as potential producer strains. These strains demonstrate a high growth rate and resistance to low temperatures in the short summer period, as well as high insolation, UV radiation, and aridity. Further studies will focus on identifying the biologically active substances, clarifying their chemical structures, and comparing them with existing compounds regarding their herbicidal activity. This will allow us to evaluate potential ecological risks and proceed to the formulation development.

Supplementary Materials

The following supporting information can be downloaded at https://www.mdpi.com/article/10.3390/agronomy13061619/s1, Figure S1: High performance liquid chromatography–mass spectrometry analysis of Phoma herbarum; Table S1: Reference Sequences; Figure S2: Gas chromatography–mass spectrometry (GC-MS) analysis of Phoma herbarum.

Author Contributions

Conceptualization, M.M.-L. and S.S.; data curation, M.M.-L.; formal analysis, E.K., D.M. and M.M.-L.; methodology E.G., A.T., A.V. and D.M.; supervision, E.K.; writing: original draft, A.T.; writing: review and editing, S.S. and D.M. All authors have read and agreed to the published version of the manuscript.

Funding

This research received no external funding.

Data Availability Statement

Not applicable.

Acknowledgments

Sequencing and chromatography was carried out using the equipment of the ‘Development of Molecular and Cellular Technologies’ Resource Center at St. Petersburg State University.

Conflicts of Interest

The authors declare no conflict of interest.

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Agronomy 13 01619 g001
Figure 1. Colonies of Phoma strains 20-A7-1.M19 (a) and 20-A7-1.M29 (b) on Czapek-Dox agar, pycnidia forming on agar surface (exhibiting 1 to 3 osteoles) (60× magnification), and pycnidia microscopic structures.
Figure 1. Colonies of Phoma strains 20-A7-1.M19 (a) and 20-A7-1.M29 (b) on Czapek-Dox agar, pycnidia forming on agar surface (exhibiting 1 to 3 osteoles) (60× magnification), and pycnidia microscopic structures.
Agronomy 13 01619 g001
Agronomy 13 01619 g002
Figure 2. Phylogenetic consensus tree based on EF1-α, TUB, and ITS sequences calculated using the maximum likelihood method. Numbers on nodes indicate bootstraps (n = 1000 replicates). There was a total of 914 positions in the final dataset.
Figure 2. Phylogenetic consensus tree based on EF1-α, TUB, and ITS sequences calculated using the maximum likelihood method. Numbers on nodes indicate bootstraps (n = 1000 replicates). There was a total of 914 positions in the final dataset.
Agronomy 13 01619 g002
Agronomy 13 01619 g003
Figure 3. Chlorotic (A2,A3) and necrotic (B2,B3) spots on the leaves of T. officinale (A) and S. canadensis (B) in the experimental variants opposite to the control (A1,B1).
Figure 3. Chlorotic (A2,A3) and necrotic (B2,B3) spots on the leaves of T. officinale (A) and S. canadensis (B) in the experimental variants opposite to the control (A1,B1).
Agronomy 13 01619 g003
Table 1. Germination of T. officinale and S. canadensis seeds after treatment with cultural liquid (CL), cell-free culture fugate (CFCF), and crude mycelium extract (CME) of P. herbarum 20-A7-1.M19 and P. herbarum 20-A7-1.M29. LSD0.05 = 9.
Table 1. Germination of T. officinale and S. canadensis seeds after treatment with cultural liquid (CL), cell-free culture fugate (CFCF), and crude mycelium extract (CME) of P. herbarum 20-A7-1.M19 and P. herbarum 20-A7-1.M29. LSD0.05 = 9.
Treatment SamplesSeed Germination, %
Taraxacum officinaleSolidago canadensis
Control (H2O)7078
Control (0.05% Ethyl acetate)6877
CL M193048
CFCF M192821
CME M191031
CL M294022
CFCF M293015
CME M291032
Table 2. Some pigments isolated from phoma-like fungi.
Table 2. Some pigments isolated from phoma-like fungi.
Metabolite ClassSubstancePhoma-like Fungi SpeciesPigment ColorUV λmax/nmMonoisotopic Mass,
g/mol		Ref.
PolyketidesPhomaliginPhoma lingam,
P. wasabiae		Bright yellow415, 330, 245311.17327290[27]
Wasabidienone AP. wasabiaeYellow(CHCl3)
406, 275, 245		268.13107373[28]
Wasabidienone B, C, EBrownish yellow311.17327290 *
HydroxyanthraquinoneCynodontin (3-methyl-1,4,5,8-tetrahydroxyanthraquinone)Setophoma terrestris (=Phoma terrestris)BronzeNo data286.04773803[29]
Pachybasin (1-hydroxy-3-methylanthracene-9,10-dione 9,10 Anthracenedione, 1-hydroxy-3-methyl- 1-Hydroxy-3-methylanthraquinone)Phoma exigua var foveata (=Phoma foveata)Yellow(EtOH)
403, 281, 252, 224		238.062994177[30]
Emodin (6-methyl-1,3,8-trihydroxyanthraquinone)Orange437, 289, 265, 252, 222270.05282342
Chrysophanol (3-Methylchrysazin 1,8-Dihydroxy-3-methylanthraquinone)Red436, 288, 278, 256, 226254.05790880
Phomarin (1,6-dihydroxy-3-methyl-9,10-anthraquinone)Orange(MeOH)
215,231, 251, 338, 356, 441
* Monoisotopic Mass for Wasabidienone E
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MDPI and ACS Style

Trigubovich, A.; Mandryk-Litvinkovich, M.; Valakhanovich, A.; Gorodilova, E.; Malygin, D.; Kalamiyets, E.; Sokornova, S. Psychrotolerant Strains of Phoma herbarum with Herbicidal Activity. Agronomy 2023, 13, 1619. https://doi.org/10.3390/agronomy13061619

AMA Style

Trigubovich A, Mandryk-Litvinkovich M, Valakhanovich A, Gorodilova E, Malygin D, Kalamiyets E, Sokornova S. Psychrotolerant Strains of Phoma herbarum with Herbicidal Activity. Agronomy. 2023; 13(6):1619. https://doi.org/10.3390/agronomy13061619

Chicago/Turabian Style

Trigubovich, Andrey, Maryna Mandryk-Litvinkovich, Anastasiya Valakhanovich, Elizaveta Gorodilova, Daniil Malygin, Emiliya Kalamiyets, and Sofia Sokornova. 2023. "Psychrotolerant Strains of Phoma herbarum with Herbicidal Activity" Agronomy 13, no. 6: 1619. https://doi.org/10.3390/agronomy13061619

APA Style

Trigubovich, A., Mandryk-Litvinkovich, M., Valakhanovich, A., Gorodilova, E., Malygin, D., Kalamiyets, E., & Sokornova, S. (2023). Psychrotolerant Strains of Phoma herbarum with Herbicidal Activity. Agronomy, 13(6), 1619. https://doi.org/10.3390/agronomy13061619

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