applied

sciences
Article
Effective Use of Flax Biomass in Biorefining Processes
Jolanta Batog , Aleksandra Wawro * , Weronika Gieparda , Krzysztof Bujnowicz, Joanna Foksowicz-Flaczyk ,
Szymon Rojewski , Magdalena Chudy and Marcin Praczyk

Institute of Natural Fibres and Medicinal Plants—National Research Institute, Wojska Polskiego 71B,
60-630 Poznań, Poland; jolanta.batog@iwnirz.pl (J.B.); weronika.gieparda@iwnirz.pl (W.G.);
krzysztof.bujnowicz@iwnirz.pl (K.B.); joanna.flaczyk@iwnirz.pl (J.F.-F.); szymon.rojewski@iwnirz.pl (S.R.);
magdalena.chudy@iwnirz.pl (M.C.); marcin.praczyk@iwnirz.pl (M.P.)
* Correspondence: aleksandra.wawro@iwnirz.pl; Tel.: +48-61-8455814

Abstract: Flax is one of the few plants that are entirely a source of raw materials for further production.
Promising directions for the use of flax biomass may be the production of bioenergy in the form
of 2G biofuels and the production of “green” composites. The aim of the study is to compare the
biomass of fiber flax, linseed and dual-purpose varieties of cultivated flax (Linum usitatissimum L.)
susceptibility to the biorefining processes. In the first stage of the research, based on the results of
yield structure features and biometric measurements of plants, the most optimal flax line was selected
for the fiber flax, linseed and dual purpose. Next, the forms of flax were pretreated with sodium
hydroxide (NaOH), the chemical composition was determined and SEM images were taken. The
obtaining of bioethanol process SFF (simultaneous saccharification and fermentation) was carried
out. In addition, biodegradable polymers were modified with flax biomass, shapes were prepared,
and the rheological and mechanical properties, as well as microbiological activity of biocomposites,
were determined. The highest concentration of ethanol (8.72 g·L−1 ) and the greatest susceptibility to
mold fungi of the biocomposites were obtained for the fiber flax variety PET 16/20.

Keywords: flax biomass; biorefining process; alkaline pretreatment; SSF; bioethanol; biocomposites;
mold fungi

Citation: Batog, J.; Wawro, A.;

Gieparda, W.; Bujnowicz, K.;
Foksowicz-Flaczyk, J.; Rojewski, S.;
1. Introduction
Chudy, M.; Praczyk, M. Effective Use
of Flax Biomass in Biorefining Cultivated flax (Linum usitatissimum L.) belongs to the agricultural species with a very
Processes. Appl. Sci. 2023, 13, 7359. wide range of applications. It is an annual plant with medicinal and dietary properties,
https://doi.org/10.3390/ also used in industry. Three types of flax are distinguished in cultivation forms: fiber flax,
app13137359 linseed and dual purpose [1]. The total cultivated area of flax in the world in 2021 was
4,384,000 ha, of which only 241,103 ha was used for fiber flax [2].
Academic Editor: In Jung Kim
Linseed varieties are focused on a high yield of seeds, which are characterized by
Received: 21 April 2023 the highest content of alpha-linolenic acid among plants [3,4]. Flax seeds contain 33–45%
Revised: 14 June 2023 fat, and linseed oil over 50% of highly saturated fatty acids—mainly alpha-linolenic acid
Accepted: 19 June 2023 (omega 3) and linoleic acid. The main producers of linseed are Russia (1,492,119 ha),
Published: 21 June 2023 Kazakhstan (1,366,068 ha) and Canada (403,500 ha) [2]. Flax seeds are used in the food and
baking industry (bread, biscuits and breakfast cereals), the oil industry (edible oil, paints,
varnishes, printing inks), the pharmaceutical industry and as dietary supplements.
The basic raw material obtained from the cultivation of fibrous forms is straw and fiber.
Copyright: © 2023 by the authors.
Flax straw is used to obtain long fiber used to produce high-quality yarns and fabrics, and
Licensee MDPI, Basel, Switzerland.
This article is an open access article
short fiber, which is used to produce carded yarns, insulating mats or paper [5,6]. Fiber flax
distributed under the terms and
is mainly cultivated in France (112,580 ha), Belarus (42,300 ha) and Russia (36,483 ha) [2].
conditions of the Creative Commons
In addition, to increase the profitability of flax cultivation, breeding works are carried
Attribution (CC BY) license (https:// out to produce dual-purpose varieties [7–9]. It is an attempt to combine a high yield of
creativecommons.org/licenses/by/ seeds with a high yield of monomorphic fiber, intended for technical purposes.
4.0/).

Appl. Sci. 2023, 13, 7359. https://doi.org/10.3390/app13137359 https://www.mdpi.com/journal/applsci

Appl. Sci. 2023, 13, 7359 2 of 15

However, the fact that natural bast fibers constitute a negligible part of the global
textile market makes it difficult to sell basic raw materials obtained from flax, especially
fiber. The main reason for this is the replacement of flax fiber with cotton and synthetic
fibers, and the high cost of natural fiber products. Moreover, weather conditions have a
great influence on the quality of the fiber. All this makes it necessary to develop alternative
directions for the use of flax biomass [10]. One of them is the use of flax biomass in
biorefining processes.
Lignocellulosic biomass is the oldest and most widespread source of renewable energy,
the third largest natural energy source in the world. The use of biomass is inextricably
linked to the reduction of greenhouse gas emissions, improvement of energy security and
socio-economic development.
The cultivation and processing of flax plants can be climate-neutral and harmoniously
integrated into a circular bioeconomy. As mentioned, thanks to the multi-directional
use of flax raw materials, by-products do not have to be waste, but a raw material for
further production.
In this study, one of the planned biorefining processes of flax biomass is the process of
obtaining 2G bioethanol, because according to the assumptions of the RED II Directive, it
introduced the obligation to reach 14% share of RES in transport by 2030, including at least
3.5% from advanced biofuels (from non-food plant matter) [11].
The structure of plant biomass is formed by lignocellulose, in which cellulose and
hemicellulose are potential substrates in the process of obtaining biofuels, but lignin
consisting of aromatic compounds is a strong obstacle in this process.
Therefore, there is a need to carry out pretreatment, the purpose of which is to loosen
the compact structure of lignocellulose and delignify the raw material [12]. The next
stages of processing plant biomass into bioethanol are enzymatic hydrolysis and ethanol
fermentation. An effective and economical solution seems to be the SSF process combining
cellulose hydrolysis with sugar fermentation in one bioreactor [13–15].
Natural plant raw materials often contain substances that have antibacterial or anti-
fungal properties, which reduces the number of microorganisms and helps to keep them at
a safe, low level.
The current market, and especially the requirements of today’s customers, prefers a
hygienic lifestyle and attaches more and more importance to the issue of health protection
and care for the natural environment. In addition, due to legal regulations, environmentally
friendly materials are increasingly sought by manufacturers of industrial products and
their users [16,17].
The plastics industry is looking for new materials that will reduce the amount of waste.
Both legislation and consumer expectations require less environmental impact from plastics.
This increases interest in environmentally friendly biodegradable materials. One of the
leading biodegradable polymers on the market is polylactic acid (PLA) in the packaging
industry [18–20].
However, in many areas, the increased use of biodegradable materials is limited by
their higher price than currently used polymers. A way to reduce the consumption of
valuable biodegradable polymers may be to replace some of them with cheaper biomass of
annual plants, e.g., from flax or hemp [21,22].
Considering that, the use of natural fillers for polymer matrices requires suitable
preparation to obtain biocomposites with good mechanical properties [23]. Increased
production of biocomposite materials in the EU will result in growing interest from industry
for components based on renewable raw materials, including flax biomass and fibers. All
this contributes to the great interest in research on the mechanism and properties of PLA
composites with natural fillings derived from various plant materials [24–27].
Overview studies on the production of bioethanol from lignocellulosic biomass and
the authors’ experience in this scope [28,29], allow us to say that the conducted research is
an important element in the search for an alternative method of flax straw management to
 to expand the sale of the raw material and increase the profitability of crops. So far, there
are no literature reports describing the use of flax biomass in the biorefining process.
Therefore, the aim of this presented study is the compared perspective lines of culti-
Appl. Sci. 2023, 13, 7359 3 of 15
vated flax—linseed (LS), fiber flax (FF) and dual purpose (DP), obtained by crossing se-
lected parental forms, in terms of the efficiency of the bioethanol production process and
the functional and microbiological properties of biocomposites with flax biomass.
expand the sale of the raw material and increase the profitability of crops. So far, there are
no literature reports describing the use of flax biomass in the biorefining process.
2. Materials and
Therefore, Methods
the aim of this presented study is the compared perspective lines of cul-
2.1. Flax flax—linseed
tivated Biomass (LS), fiber flax (FF) and dual purpose (DP), obtained by crossing
selected parental forms, in terms of the efficiency of the bioethanol production process and
The cultivated flax biomass was obtained from the Experimental Farm of the Institute
the functional and microbiological properties of biocomposites with flax biomass.
of Natural Fibres and Medicinal Plants National Research Institute in Pętkowo, Poland.
The research
2. Materials material consisted of 12 homogeneous lines of three functional forms of
and Methods
flax (Table 1).
2.1. Flax Biomass
The cultivated flax biomass was obtained from the Experimental Farm of the Institute
Table 1. ListFibres
of Natural of flaxand
lines tested in Plants
Medicinal the experiment.
National Research Institute in P˛etkowo, Poland.
The research material consisted of 12 homogeneous lines of three functional forms of
flax No.
(Table 1). Flax Lines Cultivation Form
1 PET 16/02 Linseed
Table 21. List of flax lines tested
PET 16/06
in the experiment. Linseed
3 No. PET 16/07 Flax Lines Linseed
Cultivation Form
4 1
PET 16/16
PET 16/02
Linseed
Linseed
5 2 PET 16/21 PET 16/06 Linseed
Linseed
6 3 SW 16/01 PET 16/07 Dual purpose
Linseed
4 PET 16/16 Linseed
7 5
SW 16/05 PET 16/21
Dual purpose
Linseed
8 6 SW 16/08 SW 16/01 Dual purpose
Dual purpose
9 7 SW 16/13 SW 16/05 Dual purpose
Dual purpose
8 SW 16/08 Dual purpose
10 9 PET 16/20 SW 16/13 DualFiber flax
purpose
11 10 PET 16/09 PET 16/20 Fiber
Fiber flax
flax
12 11 PET 16/23 PET 16/09 Fiber
Fiber flax
flax
12 PET 16/23 Fiber flax

The breeding lines were obtained by controlled pollination of the parental forms
then, The breeding lines were obtained by controlled pollination of the parental forms
the best single plants for reproduction were selected from the obtained genotypes.
then, the best single plants for reproduction were selected from the obtained genotypes.
Genetically stabilized lines of the F7 generation were used for the study.
Genetically stabilized lines of the F7 generation were used for the study.
The
The flax biomassobtained
flax biomass obtained forfor
thethe tests
tests waswas prepared
prepared according
according to Figure
to Figure 1. 1.

Figure
Figure 1.
1. Scheme ofthe
Scheme of theexperiment
experiment with
with thethe obtained
obtained flax biomass.
flax biomass.
Appl. Sci. 2023, 13, 7359 4 of 15

2.2. Cultivation of Flax

Plants of the tested flax lines were grown in a pot experiment in a greenhouse. Each line
was plated in five replications (5 pots). The pots were filled with soil from the experimental
field of valuation class IV b, then they were placed randomly. Standard mineral fertilization
was used for the cultivation of flax. A total of 30 seeds were sown in each pot. Seed sowing
was carried out in the second half of April 2021. Flax plants were harvested at the stage of
full seed maturity in the first half of August.

2.3. Biometric Measurements of Flax

The assessment of the traits that have the greatest impact on the biorefining processes
efficiency—straw yield, total plant yield, plant height and technical plant length, was the
basis for the selection process. Seed weight and panicle length were also determined. The
total yield was determined by weighing the harvested plants after removing the roots and
then plants were ginned to determine straw yield. To determine the total length of the
plant, it was measured from the beginning of the roots to the top of the panicle, whereas
measuring the plant from the root collar to the first branching of the panicle will allow
you to determine the technical length. A total of 20 randomly selected plants from each
repetition were used for biometric measurements (see Section 2.2). The results are the
average of a combination of 5 replicates.
In addition, in order to fully evaluate plants, for the three best lines, one from each
variety of flax, an analysis of the total fat content was carried out. Chemical analysis of
fat content was performed in a Soxhlet extractor. After grinding, samples of flax were
degreased for 3 days, ground and additionally degreased for 1 day. Each sample was
analyzed in duplicate.

2.4. Bioethanol Production Process

2.4.1. Flax Biomass Pretreatment
The flax raw material was crushed into particles of 20–40 mm and dried for 24 h
at 50–55 ◦ C. In the next stage, the biomass was ground using a cutting mill (Retsch SM-
200, Haan, Germany) into 2 mm fragments. The crushed raw material was chemical
pretreatment with 2% sodium hydroxide for 5 h at 90 ◦ C [28]. The weight ratio of NaOH to
biomass was 10:1. To neutralize, the alkaline solution of the tested biomass was filtered
and washed and dried for 24 h at 50 ◦ C. Then, using the Miller method, the amount of
released reducing sugars obtained after the alkaline pretreatment was determined [30]. The
enzyme test was performed using Flashzyme 200 (AB Enzyme, Darmstadt, Germany) at
a dose of 20 FPU·g−1 . The flax biomass was incubated at 55 ◦ C in 0.05 M citrate buffer
of pH 4.8 for 24 h. After this test, the supernatant was appropriately diluted, DNS acid
was added and the mixture was incubated in a boiling water bath for 10 min. Then, the
supernatant was cooled to room temperature and the absorbance at 530 nm was measured
(UV-VIS spectrophotometer, Jasco V-630, Pfungstadt, Germany).

2.4.2. Process of Simultaneous Saccharification and Fermentation (SSF)

The SSF process was carried out in 100 mL flasks and the total volume of flax biomass
hydrolysate was 40 mL. The prepared solution was subjected to pH adjustment to the
desired value (pH 4.8) using 1 M sulfuric acid and 1 M sodium hydroxide. Then Flashzyme
Plus 200 (AB Enzyme) enzyme was added in the amount of 20 FPU·g−1 and non-hydrated
freeze-dried yeast Saccharomyces cerevisiae was added in the dose of 0.5 g·L−1 (corresponding
to the cell concentration after inoculation of about 1 × 107 cfu·mL−1 ). The flasks (plugged
with stoppers with fermentation tubes) were incubated at 37 ◦ C for 72 h on a shaker
(200 rpm).
Appl. Sci. 2023, 13, 7359 5 of 15

2.5. Biocomposites Production Process

2.5.1. Natural Fillers from Halophyte Biomass
Samples of three forms of flax biomass were dried and ground in a Rekord A mill
(Jehmlich, Nossen, Germany) with a sieve separator with a mesh size of 0.5 mm. A sieve
analysis of the obtained natural fillers was performed using the Analysette 3 Spartan
analyzer (Fritsch, Idar-Oberstein, Germany) and their moisture content was determined
with a MA.X2.A moisture analyzer (Radwag, Radom, Poland).

2.5.2. Polymer Matrix

A biodegradable polymer—poly(lactic acid) of the Ingeo 3251D variety (Nature Works,
Plymouth, MN, USA) with a density of 1.24 g·mL−1 and a melt flow rate (MFR) of
35 g/10 min (190 ◦ C, 2.16 kg) was used as the matrix for the biocomposites.

2.5.3. Composites Preparation

PLA blends with 20% biomass content of flax varieties designated as PLA-FF-20,
PLA-DP-20 and PLA-LS-20 were compounded in co-rotating twin screw extruder Leistritz
MICRO 27 GL/GG-44D (Leistritz Extrusionstechnik, Nürnberg, Germany) with Brabender
gravimetric feeding system (Brabender Technologie, Duisburg, Germany). Compounding
conditions: barrel temperature profile 170–200 ◦ C, extrusion speed of 150 rpm and through-
put 16 kg·h−1 . Then, the composite granules were dried to a humidity level below 0.05% in
a Drywell DW25/40 molecular dryer (Digicolor, Herford, Germany) at a temperature of
70 ◦ C (dew point −40 ◦ C).
In order to determine the properties of the obtained biocomposites and microbiological
tests, type 5A test samples in accordance with ISO 527-2 were molded by injection molding
machine Haitian Mars MA600 (Haitian Plastics Machinery, Ningbo, China) [31]. Tempera-
ture profiles were 180 ◦ C (hopper), 185 ◦ C, 190 ◦ C and 190 ◦ C (nozzle). The temperature of
the mold was set to 40 ◦ C.

2.5.4. Resistance of Biocomposites to Mold Fungi

The samples of polylactide biocomposites with 20% biomass content from three vari-
eties of flax: PLA-LS-20, PLA-DP-20 and PLA-FF-20 and samples of pure polylactide—PLA
3251D as references were used for the test of resistance to mold fungi. The tests were
carried out in accordance with the methodology specified in the standard ISO 846 [32].
The tested samples were allocated on an agar medium and inoculated with a mixture
of five strains of mold fungi, which most often cause the decomposition of cellulose:
Aspergillus niger, Chaetomium globosum, Penicillium ochrochloron, Paecilomyces variotii and
Penicillium funiculosum. The agar medium with mineral salts was used, prepared in accor-
dance with the standard, using Chempur reagents and agar powder from Merck, whereas
tested microorganisms originated from the pure culture collection of the Institute of Fer-
mentation Technology and Microbiology, Technical University of Lodz, Poland.
Then, the inoculated samples were placed in a climatic chamber in optimal conditions
for mold development, i.e., at a temperature of 29 ± 1 ◦ C and relative air humidity of
90% for 28 days. These samples were designated as series I. For comparison, samples of
biocomposites in each variant, which were allocated on a sterile agar medium and not
inoculated with a mixture of mold fungi, were placed in the same climatic conditions. These
samples were designated as series S. In the case of samples not inoculated with the test
fungi, the degree of infection with microorganisms other than the test fungi was assessed.
In addition, biocomposite control samples for each variant, not incubated, were left and
stored under standardized conditions of room temperature and relative humidity. These
samples were designated as series 0.
After 28 days, visual assessment of fungal growth on the samples’ surfaces was
assessed on a scale from 0 to 5, where:
0—no visible growth under the microscope
1—growth invisible to the naked eye but clearly visible under the microscope
Appl. Sci. 2023, 13, 7359 6 of 15

2—growth visible to the naked eye, covering up to 25% of tested surface

3—growth visible to the naked eye, covering up to 50% of tested surface
4—considerable growth, covering more than 50% of tested surface
5—intensive growth, covering all tested surface.
Next, the samples were cleansed and mechanical properties were evaluated. All the
measurements were performed six times.

2.6. Analytical and Testing Methods

The chemical composition of the main cultivated flax biomass components before and
after alkaline pretreatment was determined according to standard methods. The cellulose con-
tent was determined by the Seifert method using a mixture of acetylacetone and dioxane [33].
The lignin content was determined by the Tappi (Technical Association of the Pulp and Pa-
per Industry) method (T-222 om-06) using concentrated sulfuric acid [34]. The theoretical
hemicellulose content was arithmetically calculated as the difference between holocellulose
(TAPPI—T9 wd-75) and cellulose [35]. All chemical composition results were the average of
three measurements and were calculated on the basis of the dry weight of the raw material.
In addition, for flax biomass before and after pretreatment, using a scanning electron
microscope (SEM, S-3400N, Hitachi, Tokyo, Japan) under high vacuum conditions, the
physical morphology was determined. Biomass samples were covered with gold dust.
The ethanol concentration from flax biomass was determined by HPLC on Elite
LaChrom liquid chromatograph from VWR-Hitachi using an RI L-2490 detector, Rezex
ROA 300 × 7.80 mm column from Phenomenex, at a flow rate of 0.6 mL·min−1 , at 40 ◦ C.
The samples were loaded onto the column at 10 µL. The quantitative identification was per-
formed by the external standard method using the peak area (measurement and computer
integration using the Ez-Chrom Elite program).
Mechanical properties tests of the composites were carried out at room temperature
with a universal testing machine Inspekt Table 50 (Hegewald & Peschke MPT, Nossen,
Germany) according to ISO 527 and ISO 178, respectively [31,36,37]. In both tests, the
crosshead speed was set to 5 mm·min−1 . Tensile tests in accordance with ISO 527-1 and
2 were carried out using an MFA clip-on extensometer (MF Mess- & Feinwerktechnik,
Velbert, Germany) with a nominal length of 20 mm [31,36].

2.7. Statistical Analysis

The SSF process experiments were performed in triplicate. Standard ethanol concentra-
tion deviations were calculated using ANOVA analysis of variance, Statistica 13.0 software
(p < 0.05) [38].

3. Results and Discussion

3.1. Selection of Optimal Genotype for the Fiber Flax, Linseed and Dual Purpose
A comparative analysis of selected lines of flax with a high degree of homozygosity
was carried out in order to select the best genotypes for biorefining processes.
The lines were compared in terms of the following yield-forming features: total yield,
straw yield, seed yield, plant height, technical length and panicle length. The test results
are presented in Tables 2–4.

Table 2. Comparison of tested linseed lines in terms of yield structure traits.

Total Straw Seed Plant Technical Panicle

Breeding Yield Yield Yield Height Length Length
Line
(g) (g) (g) (cm) (cm) (cm)
PET 16/02 146.4 ± 47.3 76.4 ± 25.6 48.0 ± 11.4 50.5 ± 3.1 39.3 ± 2.0 11.2 ± 2.5
PET 16/06 126.4 ± 22.5 52.8 ± 5.2 31.6 ± 9.8 46.3 ± 2.5 36.6 ± 1.9 9.7 ± 2.9
PET 16/07 92.8 ± 21.9 32.4 ± 17.7 31.6 ± 9.8 42.5 ± 2.8 33.9 ± 1.5 8.6 ± 1.5
PET 16/16 200.4 ± 16.6 79.6 ± 17.8 70.0 ± 4.5 50.0 ± 2.7 40.5 ± 2.8 9.4 ± 1.4
PET 16/21 111.2 ± 28.2 48.8 ± 11.5 28.8 ± 9.9 47.6 ± 5.1 36.3 ± 4.6 11.2 ± 1.7
Appl. Sci. 2023, 13, 7359 7 of 15

A significant differentiation of the tested linseed lines was found in terms of yield-
forming features, i.e., total yield, straw yield and seed yield. The PET 16/16 line was
characterized by significantly the highest total plant weight and seed weight compared
to the other tested linseed genotypes. The straw weight of this line was also the highest,
with significant differences observed in comparison to the lines: PET 16/06, PET 16/07 and
PET 16/21.
There was definitely less differentiation between the examined objects observed in the
case of morphological traits, i.e., plant height, technical length and panicle length.
For biomass biorefining processes, however, straw yield and total yield are the most
important, and on this basis, the best line was PET 16/16.

Table 3. Comparison of tested dual-purpose lines in terms of yield structure traits.

Total Straw Seed Plant Technical Panicle

Breeding Yield Yield Yield Height Length Length
Line
(g) (g) (g) (cm) (cm) (cm)
SW 16/01 153.6 ± 25.3 57.2 ± 16.2 52.8 ± 14.8 43.7 ± 8.6 33.4 ± 7.5 10.0 ± 1.9
SW 16/05 123.2 ± 41.7 48.8 ± 18.0 48.0 ± 16.0 39.4 ± 3.6 29.7 ± 3.2 9.6 ± 1.4
SW 16/08 120.8 ± 23.8 50.8 ± 14.3 38.8 ± 5.4 46.0 ± 2.0 38.0 ± 2.4 8.0 ± 1.7
SW 16/13 143.6 ± 19.9 64.0 ± 14.6 48.8 ± 8.3 44.6 ± 6.6 35.0 ± 4.0 9.5 ± 2.6

The differentiation of the tested dual-purpose lines in terms of yield-producing traits

was lower than in the case of linseed lines. The highest total plant weight was found in
line SW 16/01, whereas the highest mass of straw was characterized by line SW 16/13.
The straw weight of the SW 16/13 line was 10.6% higher than the straw weight of the SW
16/01 line, with the total weight of the plants being lower by 6.6%. However, since the
mass of straw is considered to be the most important trait for biomass biorefining processes,
a line SW 16/13 was selected for further chemical research.

Table 4. Comparison of tested fiber flax lines in terms of yield structure traits.

Total Straw Seed Plant Technical Panicle

Breeding Yield Yield Yield Height Length Length
Line
(g) (g) (g) (cm) (cm) (cm)
PET 16/20 122.8 ± 14.3 70.4 ± 8.5 36.8 ± 3.3 50.9 ± 3.0 41.7 ± 1.7 9.2 ± 1.6
PET 16/09 113.2 ± 28.2 70.0 ± 22.9 28.4 ± 8.7 67.6 ± 5.2 50.4 ± 5.4 17.2 ± 1.3
PET 16/23 125.2 ± 10.1 66.8 ± 11.5 37.6 ± 6.2 53.4 ± 3.2 41.2 ± 4.1 12.2 ± 2.6

Within fiber flax lines, very slight differences in the tested objects in terms of straw
weight were observed. Therefore, the most important trait did not differentiate the com-
pared lines to a sufficient extent to make a clear choice of the best one on this basis.
The highest straw weight was found for line PET 16/20, but line PET 16/09 showed
only 0.4 g less weight of straw. However, the line that showed the highest straw yield (PET
16/20) was also characterized by a slightly lower total yield than the best line (PET 16/23),
and significantly higher than the line PET 16/09. On this basis, it was decided that the line
PET 16/20 would be directed for further chemical research.
Summing up, in this study, the straw yield was significantly highest among the fiber
flax lines (an average of 69 g), and the lowest within the dual-purpose lines (an average
of 55.2 g). However, dual-purpose lines were characterized by a slightly lower mass of
straw than linseed lines (an average of 58 g). In turn, the seed yield was the highest
in dual-purpose forms, slightly lower in linseed forms, and significantly lowest in fiber
flax forms.
Since it is possible to effectively use both straw and seeds in the cultivation of flax, the
total fat content in the seeds of three selected lines of flax was also analyzed (Table 5).
 Since it is possible to effectively use both straw and seeds in the cultivation of flax,
the total fat content in the seeds of three selected lines of flax was also analyzed (Table 5).

Appl. Sci. 2023, 13, 7359 Table 5. Comparison of selected lines of flax in terms of fat content in seeds. 8 of 15

No. Breeding Line Fat Content (%)

1 PET 16/16 30.9 ± 0.4
Table 5. Comparison of selected lines of flax in terms of fat content in seeds.
2 SW 16/13 31.1 ± 0.6
3 No. PET 16/20Breeding Line Fat28.5 ± 0.3(%)
Content
1 PET 16/16 30.9 ± 0.4
The highest2 fat content was foundSW in the dual-purpose line SW
16/13 16/13.
31.1 ± 0.6 In this respect,
3 PET 16/20 28.5 ± 0.3
it exceeded not only the fiber flax form (PET 16/20) but also the linseed form (PET 16/16).
Whereas, fat content in the seeds of linseed forms is higher compared to fiber flax forms,
which is The highest fatby
confirmed content was found
numerous in the
studies dual-purpose
[39,40]. However,line SW
the 16/13. In between
difference this respect,
the best
it exceeded not only the fiber flax form (PET 16/20) but also the linseed form (PET 16/16).
line SW 16/13 and the worst line PET 16/20 was only 2.6%. This proves the high lability of
Whereas, fat content in the seeds of linseed forms is higher compared to fiber flax forms,
thewhich
fat content in flaxbyseeds
is confirmed and the
numerous possibility
studies of profitable
[39,40]. However, use of seeds,
the difference regardless
between the bestof the
cultivation form.
line SW 16/13 and the worst line PET 16/20 was only 2.6%. This proves the high lability of
the fat content in flax seeds and the possibility of profitable use of seeds, regardless of the
3.2.cultivation
Bioethanolform.
Production Process
3.2.1. Flax Biomass Pretreatment
3.2. Bioethanol Production Process
Physicochemical
3.2.1. pretreatment was carried out, which is a very important stage in the
Flax Biomass Pretreatment
conversion of biomass for
Physicochemical energy purposes.
pretreatment Theout,
was carried effect of the
which is aprocess is to loosen
very important stagethe
in com-
pact
thestructure
conversion ofoflignocellulose,
biomass for energywhich enablesThe
purposes. better access
effect of theof enzymes
process is to to cellulose,
loosen the in-
creasing
compactthestructure
susceptibility of biomasswhich
of lignocellulose, to decomposition
enables better into
access simple sugars,
of enzymes towhich are then
cellulose,
converted into
increasing theethanol in theof
susceptibility fermentation process.
biomass to decomposition into simple sugars, which are
The
then selectedinto
converted flaxethanol
line’s biomass was ground
in the fermentation using a knife mill on a sieve with a mesh
process.
of 2 mm. The biomass was treated with 2% sodium
The selected flax line’s biomass was ground usinghydroxide
a knife milland
on athe content
sieve with a of released
mesh
of 2 mm. The biomass was treated with 2% sodium hydroxide and the content of released
reducing sugars was determined using the Miller method (Figure 2).
reducing sugars was determined using the Miller method (Figure 2).

Reducing sugars (mg∙g-1)

260
240
220
200
Dual Linseed Fiber flax
purpose

Figure 2. Content of reducing sugars in flax biomass after pretreatment.

Figure 2. Content of reducing sugars in flax biomass after pretreatment.
It was found that the highest content of reducing sugars was obtained for the form of
It was
fiber found
flax—the linethat
PETthe highest
16/20, content
which of reducing
amounted to 248.1 mgsugars
·g− 1 . was obtained
The linseed for the
turned form of
out to
fiber flax—the line PET 16/20, which amounted to 248.1 mg·g −1. The linseed turned out to
be the most alkali resistant. However, the difference between these forms was not too large
be and
the most alkalitoresistant.
amounted 27.3 mg·gHowever, the difference between these forms was not too large
− 1 , which was 11% of the value obtained for fiber flax.

and amounted
Next, the to 27.3 mg·g
chemical −1, whichof
composition was 11%
three of theofvalue
varieties obtained
flax biomass wasfor fiber flax.
assessed, before
andNext, the chemical composition of three varieties of flax biomass was assessed, before
after alkaline treatment using chemical methods (Table 6).
and after alkaline treatment using chemical methods (Table 6).
Table 6. Chemical composition of flax biomass before (BP) and after (AP) alkaline treatment (%).

TableCultivation
6. Chemical composition of flax biomass before (BP) and after (AP) alkaline treatment (%).
Sample Cellulose Holocellulose Hemicellulose Lignin
Form
Cultivation
Sample
BP Cellulose
37.42 ± 0.62 Holocellulose
69.31 ± 0.99 Hemicellulose
31.89 ± 0.41 Lignin
17.83 ± 0.43
Form
Linseed
AP 47.82 ± 0.17 71.61 ± 0.39 23.79 ± 0.51 20.30 ± 0.60
BPBP 37.42 ± 0.62
± 0.21 69.31
72.13 ±±0.99 31.89 ± 0.4118.44 ±17.83
± 0.42 0.10 ± 0.43
Linseed
Dual purpose
39.94 0.26 32.19
APAP 47.82 ± 0.17
49.20 ± 0.56 71.61 ±±0.39
72.31 0.44 23.79 ± 0.51
23.11 ± 0.54 21.26 ±20.30
0.04 ± 0.60
Dual purpose BPBP 39.94 ± 0.21
42.17 ± 0.29 72.13
74.27 ±±0.26
0.47 32.19
32.10 ± 0.4216.93 ±18.44
± 0.63 0.31 ± 0.10
Fiber flax
AP 51.40 ± 0.58 75.72 ± 0.21 24.32 ± 0.55 19.86 ± 0.30
Appl. Sci. 2023, 13, 7359 9 of 15

The fiber flax has the highest cellulose content, which is confirmed by the results
of the content of reducing sugars. Alkaline treatment resulted in partial degradation
of hemicellulose. The seeming increase in lignin content was mainly due to the loss of
hemicellulose as a result of the alkaline pretreatment environment.
Similar values of chemical composition were observed for the biomass of other plants
such as hemp. Before chemical treatment, cellulose was in the range of 47–51%, hemicel-
lulose was equal to 28–33% and lignin was 14–16%. Whereas, hemp biomass subjected
to chemical treatment with NaOH was characterized by an increased content of cellulose
(57–63%), as well as partial degradation of hemicellulose (20–22%). In turn, the lignin
content, similarly to flax biomass, slightly increased after chemical treatment (17–18%) [29].
On the other hand, for sorghum biomass treated with 2% sodium hydroxide, an increase in
the cellulose content from 37% (before pretreatment) to 71% was observed, and a decrease
in the content of hemicellulose from 25% to 16%, and especially lignin from 21% to 6% [41].
SEM photos of biomass were also taken for all forms of flax before and after treatment
with 2% NaOH (Table 7).

Table 7. SEM photos of flax biomass before and after pretreatment.

Before Pretreatment After Treatment with 2% NaOH

Dual purpose

Linseed

Fiber flax

Untreated flax biomass is intact and has a sedimentary layer on the surface that
effectively blocks access to lignocellulose [42]. After treatment with NaOH, all tested
varieties of flax, especially fiber flax and dual purpose, show damage to the biomass
structure and partial cleaning of its surface [43]. This proves the effectiveness of the alkaline
treatment and has a positive effect on the enzymatic availability and digestibility of biomass
in the subsequent stages of the bioethanol production process.
Appl. Sci. 2023, 13, x FOR PEER REVIEW 10 of 16

and partial cleaning of its surface [43]. This proves the effectiveness of the alkaline treat-
Appl. Sci. 2023, 13, 7359 ment and has a positive effect on the enzymatic availability and digestibility of biomass
10 of 15
in the subsequent stages of the bioethanol production process.

3.2.2.
3.2.2.SSF
SSFProcess
ProcessofofFlax
FlaxBiomass
Biomass
The
The biomass of selectedflax
biomass of selected flaxlines
lineswas
wassubjected
subjectedtotothe
theSSF
SSFprocess
processfor
for72
72hhatat37
37°C,
◦ C,
i.e., in conditions appropriate for the synergy of the Flashzyme Plus 200 enzyme
i.e., in conditions appropriate for the synergy of the Flashzyme Plus 200 enzyme and and Sac-
charomyces cerevisiae
Saccharomyces distillery
cerevisiae yeast.
distillery Figure
yeast. 3 shows
Figure the ethanol
3 shows concentration
the ethanol determined
concentration deter-
by HPLC.
mined by HPLC.

Figure
Figure3.3.Ethanol
Ethanolconcentration
concentrationof
offlax
flaxbiomass:
biomass:LS—linseed;
LS—linseed;DP—dual
DP—dualpurpose;
purpose;FF—fiber
FF—fiberflax.
flax.

Thehighest
The highest ethanol
ethanol content
content was
was obtained
obtained for
for fiber
fiber flax
flax biomass
biomass and
and it
it was
was 8.72
8.72 gg·L·L− 1
−1, ,
andthe
thelowest
lowest for for linseed
linseed was
was 7.65
7.65 g·L − 1
g·L−1 (a(adifference
differenceofofabout
about12%
12%compared
comparedto tothethevalue
value
and −1 ) was
of fiber flax). Moreover,
of fiber flax). Moreover, the difference between linseed and dual purpose (8.18
the difference between linseed and dual purpose (8.18 g·L ) was g · L −1
about0.5
about g·L−1−, 1the
0.5g·L , the same
same asas between
between dual
dual purposeand
purpose andfiber
fiberflax.
flax.
A similar ethanol concentration was obtained for hemp biomass, which was pretreated
A similar ethanol concentration was obtained for hemp biomass, which was pre-
with sodium hydroxide of the same concentration, i.e., 2%, and then the SSF process with
treated with sodium hydroxide of the same concentration, i.e., 2%, and then the SSF pro-
the addition of the Flashzyme Plus 200 enzyme. The ethanol content for hemp biomass was
cess with the addition of the Flashzyme Plus 200 enzyme. The ethanol content for hemp
in the range of 6–7 g·L−1 [29]. In other studies the SSF was carried out on the pretreated
biomass was in the range of 6–7 g·L−1 [29]. In other studies the SSF was carried out−on the
Miscanthus species and the ethanol concentration from this biomass was 7.42 g·L 1 [44].
pretreated Miscanthus species and the ethanol concentration from this biomass was 7.42
Summing up, the obtained ethanol concentrations for flax biomass are at a satisfactory
g·L−1 [44].
level, and the biomass of each selected line of flax can be a potential raw material for
Summing up, the obtained ethanol concentrations for flax biomass are at a satisfac-
obtaining bioethanol.
tory level, and the biomass of each selected line of flax can be a potential raw material for
obtaining bioethanol.
3.3. Biocomposite Production Process
3.3.1. Fillers from Flax Biomass
3.3. Biocomposite Production Process
Natural fillers from flax biomass with particles smaller than 0.5 mm were prepared.
3.3.1. Fillers from
The humidity wasFlax Biomass and a sieve analysis of the natural fillers was performed.
determined
The Natural
details arefillers
shownfrominflax biomass
Table 8. with particles smaller than 0.5 mm were prepared.
The humidity was determined and a sieve analysis of the natural fillers was performed.
The
Tabledetails are shown
8. Particle in Table and
size distribution 8. humidity of fillers from flax biomass.
Particle Size Distribution (%)
Table 8. Particle sizeHumidity
Cultivation distribution and humidity of fillers from flax biomass.
Form (%) Below Below Below Below
0.5 mm Particle
0.25 mmSize Distribution
0.2 mm (%) 0.1 mm
Cultivation Humidity
FiberForm
flax 6.8 (%) 37 Below Below
35 Below
23 Below
5
Dual purpose 6.5 31 0.5 mm 0.25
27 mm 0.2
35 mm 0.1 7mm
Fiber flax
Linseed 7.2 6.8 28 37 33 35 3023 95

The bulk density of fillers from selected forms of flax was also determined and the
following values were obtained, i.e., for fiber flax 0.165 kg·L− 1 , dual purpose 0.172 kg·L− 1
and linseed 0.183 kg·L− 1 .
 Appl. Sci. 2023, 13, 7359 11 of 15

3.3.2. Mechanical Properties of Biocomposites

The effects of the selected flax varieties biomass on both the tensile and flexural
properties of PLA composites are shown in Table 9.

Table 9. Tensile and flexural properties of PLA/flax composites.

Tensile Strength Tensile Modulus Flexural Strength Flexural Modulus

Sample
δM (MPa) Et (GPa) δfM (MPa) Ef (GPa)
PLA 3251D 65.5 ± 1.3 3.3 ± 0.2 105.4 ± 1.0 3.4 ± 0.1
PLA-FF-20 55.6 ± 1.0 5.3 ± 0.3 89.2 ± 2.4 5.6 ± 0.2
PLA-DP-20 54.2 ± 1.2 5.1 ± 0.3 85.7 ± 1.8 4.9 ±0.3
PLA-LS-20 53.1 ± 1.1 5.1 ± 0.3 84.4 ± 1.8 4.6 ± 0.2

The use of the PLA composites of flax fillers in the amount of 20% resulted in a
decrease in tensile strength in the range of 15–19%. Compared to PLA, the flexural strength
of the composites decreased in the range of 13–19%. On the other hand, depending on the
variety of linen, an increase in modulus was noted during the elongation (54–60%) and
bending (35–64%) of the compositions in relation to PLA.
Natural raw materials containing cellulose, hemicellulose, lignins and pectins are
active hydrophilic fillers [45]. The decrease in tensile and flexural strength indicates that
despite the use of hydrophilic flax biomass in combination with a hydrophilic biodegradable
polymer (PLA), the structure of the biocomposite does not show full adhesion at the
interface of individual components [46]. The reason for this may be the different content of
fiber and shives in the biomass unit or the different content of cellulose in the composition
for different varieties of flax, which is responsible for the effectiveness of adhesion with the
biodegradable polymer matrix. The filler/matrix adhesion is an important factor affecting
the final mechanical properties of composites. Good interface adhesion ensures effective
stress transfer between the matrix and natural filler [27,47].

3.3.3. Microbiological Properties of Biocomposites

Appl. Sci. 2023, 13, x FOR PEER REVIEW 12 of
In the mold fungal test, regardless of the type of linen biomass, the growth of 16
test
fungi was observed on the surface of the tested samples inoculated with microorganisms
(series I). The results of the test are shown in Figure 4.

Intensity of mould fungi growth

PLA-LS-20

PLA-DP-20

PLA-FF-20

PLA 3251D

0 1 2 3 4 5

Series S Series I

Figure 4. Intensity of mold fungi growth on the samples’ surfaces.

Figure 4. Intensity of mold fungi growth on the samples’ surfaces.
The highest mold fungi resistance was found for biocomposites PLA-LS-20—fungal
The highest
growth mold fungi
was insignificant resistance
(second wasinfound
degree), for of
the form biocomposites PLA-LS-20—fungal
single traces of fungi on the surface
growth
of thewas insignificant
samples. (second degree),
Biocomposites PLA-FF-20in the form
were of most
the singlesusceptible
traces of fungi on test
to the the sur-
fungi.
face of the samples.
Intensive Biocomposites
fungal growth PLA-FF-20
(fourth degree) were the most
was observed on thesusceptible to the test
samples, covering fungi.
more than
Intensive
50% of fungal growth
the tested (fourth
surface. Ondegree) was observed
the samples of PLAon the samples,
composites covering
without themore thanof
addition
50% of the (PLA
biomass tested3251D),
surface.inoculated
On the samples of PLAa slight
with molds, composites without
increase the addition
was found (secondof bio-
degree),
mass (PLA up
covering 3251D),
to 25%inoculated with
of the tested molds, a slight increase was found (second degree),
surface.
covering up to 25% of the tested surface.
Composites not inoculated with molds (series S), both PLA-LS-20, PLA-DP-20 and
PLA-FF-20 showed susceptibility to microorganisms other than test fungi in the second
degree, which indicates the presence of substances that are a medium for the growth of
Appl. Sci. 2023, 13, 7359 12 of 15

Composites not inoculated with molds (series S), both PLA-LS-20, PLA-DP-20 and
PLA-FF-20 showed susceptibility to microorganisms other than test fungi in the second
degree, which indicates the presence of substances that are a medium for the growth of
microorganisms. The samples of biocomposites PLA-FF-20 were the most susceptible, and
the PLA-LS-20 were the least susceptible; however, for all samples it was growth visible
to the naked eye, covering up to 25% of the tested surface. At the same time, on samples
of PLA composites without the addition of biomass (PLA 3251D), not inoculated, trace
development of microorganisms other than test fungi was observed, visible only under
a microscope.
In addition, changes in the appearance of the tested samples were assessed. Discol-
oration of the tested samples of biocomposites containing flax biomass was found, whereas
distortions and losses on the surface of the samples were not observed.
The conducted microbiological tests showed a greater susceptibility of biocomposites
containing flax biomass to the action of mold fungi, in relation to pure PLA. Moreover, in
other studies, it was shown that the addition of lignocellulosic fillers, such as conifer bark
or coniferous sawdust to PLA composites increased the susceptibility to the mold fungi
fouling process [48]. The observed differences are related to the varieties of flax from which
the biomass was obtained. Biocomposites PLA-FF-20 were the most susceptible to the test
fungi, whereas biocomposites PLA-LS-20 were the most resistant to mold fungi. This may
be due to the fact that lignification is much stronger in the stalks of linseed varieties than in
the stalks of flax cultivated mainly for fiber.

3.3.4. Mechanical Properties of Biocomposites after Test Mold Resistance

In order to determine the effect of mold fungi on the mechanical properties of the
tested biocomposites, tensile and flexural strength tests were carried out again after the
completion of the fungal tests. The results of individual studies are presented in Table 10.

Table 10. Tensile and flexural properties of PLA/flax composites after microbiological testing:
O—reference samples; I—samples inoculated with a mixture of mold fungi; S—not inoculated
samples subjected to the same climatic conditions as the inoculated samples.

Tensile Strength Tensile Modulus Flexural Strength Flexural Modulus

Sample
δM (MPa) Et (GPa) δfM (MPa) Ef (GPa)
PLA 3251D (O) 65.5 ± 1.1 3.3 ± 0.2 105.4 ± 1.0 3.4 ± 0.1
PLA 3251D (I) 64.8 ± 1.3 3.3 ± 1.0 104.2 ± 1.4 3.4 ± 0.5
PLA 3251D (S) 65.4 ± 1.2 3.3 ± 0.6 105.2 ± 1.6 3.4 ± 0.7
PLA-FF-20 (O) 55.6 ± 1.0 5.3 ± 0.3 89.2 ± 2.4 5.6 ± 0.2
PLA-FF-20 (I) 52.4 ± 1.9 5.0 ± 0.7 84.1 ± 2.3 5.3 ± 0.8
PLA-FF-20 (S) 55.2 ± 1.8 5.3 ± 0.9 88.8 ± 2.0 5.6 ± 0.6
PLA-DP-20 (O) 54.2 ± 1.2 5.1 ± 0.3 85.7 ± 1.8 4.9 ± 0.3
PLA-DP-20 (I) 51.5 ± 2.1 4.8 ± 0.7 81.5 ± 1.7 4.7 ± 0.8
PLA-DP-20 (S) 53.8 ± 2.1 5.0 ± 1.0 85.1 ± 1.6 4.8 ± 0.8
PLA-LS-20 (O) 53.1 ± 1.1 5.1 ± 0.3 84.4 ± 1.8 4.6 ± 0.2
PLA-LS-20 (I) 51.2 ± 1.1 4.9 ± 0.6 81.4 ± 2.1 4.4 ± 1.0
PLA-LS-20 (S) 52.8 ± 2.0 5.1 ± 0.9 84.0 ± 1.7 4.6 ± 0.9

The strength of flax biomass biocomposites treated with mold over the test period
(28 days) decreased in the range of 3.5–7% compared to reference samples. The effect of
only climatic conditions on the biocomposite samples did not significantly affect the change
in mechanical properties—the changes did not exceed 1%.
Analogous changes in the strength of individual biocomposites can be observed in
terms of elastic modules. Samples treated with mold mixtures showed changes in the
elastic module in the range of 4–6% compared to reference samples. Changes in the elastic
module of non-inoculated samples did not exceed 1.2%.
Regarding the changes in mechanical properties of biocomposites with biomass of
different forms of flax, it can be noted that the largest changes are shown in samples with
Appl. Sci. 2023, 13, 7359 13 of 15

biomass of fiber flax. These changes relate to both the tensile strength and elastic module of
the biocomposites. The smallest changes in strength are observed for biocomposite samples
with linseed biomass.

4. Conclusions
In this study, five lines of linseed, four lines of dual purpose and three lines of fiber
flax were analyzed, and based on the biometric measurements, one genotype for each
form of flax was selected, i.e., linseed—PET 16/16, dual purpose—SW 16/13 and fiber
flax—PET 16/20. The obtained values of straw weights for all three forms of flax testify to
the significant potential of selected genotypes to produce a large amount of raw material
for biorefining processes.
In the next stage, the variety of flax with the greatest energy potential was selected. It
was found, based on the content of reducing sugars obtained after alkaline pretreatment,
that all selected forms of flax can be an effective raw material for bioethanol production,
especially fiber flax, for which 8.72 g·L−1 of ethanol was obtained. These results indicate
the possibility of expanding the raw material base to produce 2G bioethanol.
Whereas, the biomass fillers of the three tested varieties of flax caused a decrease in
the biocomposite’s tensile strength by several percent compared to the biodegradable PLA
polymer and a significant increase in the modulus of elasticity in the range of 53–60%. In
addition, the susceptibility of biocomposites containing flax biomass to the action of mold
fungi in relation to pure PLA was demonstrated. Biocomposites with fiber flax biomass
were the most susceptible to the tested fungi and showed the greatest changes in mechanical
properties. The lowest susceptibility to test fungi and the smallest changes in strength were
observed for samples of biocomposites with linseed biomass. The obtained results can be
the basis for the design of modern biocomposites with the assumed properties depending
on the target application, e.g., packaging for food products (vegetables, fruit), elements of
equipment for public transport vehicles and elements of furniture and interior design used
in public places.
On the basis of the conducted research, the possibility of using flax biomass, in
particular fiber flax biomass, for valuable bioproducts—bioethanol and “green” composites,
and for each tested form of cultivated flax, the possibility of profitable use of the collected
seeds was demonstrated.

Author Contributions: Conceptualization, J.B., A.W., M.P., K.B. and J.F.-F.; Methodology, J.B., A.W.,
M.P., K.B. and J.F.-F.; Software, A.W., W.G.; Validation, J.B., A.W., M.P., K.B. and J.F.-F.; Formal analysis,
J.B.; Investigation, A.W., W.G., M.C., K.B., S.R. and J.F.-F.; Resources, J.B., A.W., W.G., M.P., M.C., K.B.,
S.R. and J.F.-F.; Data curation, J.B., A.W., W.G., M.P., M.C., K.B., S.R. and J.F.-F.; Writing—original
draft preparation, J.B., A.W., W.G., M.P., K.B. and J.F.-F.; Writing—review and editing, J.B. and A.W.;
Visualization, J.B.; Supervision, J.B.; Project administration, J.B. and M.P.; Funding acquisition, J.B.
and M.P. All authors have read and agreed to the published version of the manuscript.
Funding: This research was funded by the Polish Ministry of Agriculture and Rural Development as
part of the grant awarded to the Institute of Natural Fibres and Medicinal Plants in 2022/23.
Institutional Review Board Statement: Not applicable.
Informed Consent Statement: Not applicable.
Data Availability Statement: Not applicable.
Acknowledgments: The study was conducted as a research project—Grant: Area 2. Assessment of
the potential of fiber and herbal plants in terms of their applicability in various areas of bioeconomy.
Task 2.1. Assessment of the application potential of flax and hemp according to the principle of
cascade use of fiber plants. Ministry of Agriculture and Rural Development, Poland (2021); research
project—Production of initial materials for breeding of flax new varieties with improved quality and
increased resistance to Fusarium spp. and drought. Ministry of Agriculture and Rural Development,
Poland (2022–2023).
Conflicts of Interest: The authors declare no conflict of interest.
Appl. Sci. 2023, 13, 7359 14 of 15

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