Analiza biometryczna wybranych odmian lnu oleistego o zróżnicowanej aktywności wiązania kadmu z gleby

Marcin Praczyk, Krzysztof Heller, Grażyna Silska, Przemysław Baraniecki, Karolina Wielgus, Joanna Makowiecka

Institute of Natural Fibres and Medicinal Plants, Poznań, Poland

Autor korespondencyjny – M. Praczyk, e-mail: marcin.praczyk@iwnirz.pl DOI: 10.5604/12338273.1142899

Biometric analysis of selected linseed varieties

in terms of different ability

of cadmium absorption from soil

Analiza biometryczna wybranych odmian lnu oleistego

o zróżnicowanej aktywności wiązania kadmu z gleby

Abstract

The objective of the study was an evaluation of 7 linseed cultivars from the Polish Linum Collection. Thise cultivars were characterized by different ability of cadmium absorption from soil. The examined cultivars were tested for 4 quantitative traits. The impact of the increase of cadmium content in the soil on tested traits was also examined. Multivariate analysis of variance was used to determine the relationship between tested cultivars, cadmium content in the soil and year of the experiment. Significant influence of cultivars and tested interactions for the analyzed quantitative traits was found. Total yield of ACMC Duff cultivar was significantly reduced by increasing of cadmium content in the soil to 4 mg/kg, and showed significant influence on the panicle length of tested cultivars. Calculated variability coefficients showed slight variability of studied traits, regardless of the year of experiment, as well as the concentration of cadmium in the soil. Presented results indicate that the increase of cadmium content in the soil to 4 mg/kg may result in significantly negative impact on the ontogenesis of plants of cultivated flax.

Słowa kluczowe: wpływ kadmu, ontogeneza lnu, cechy ilościowe Streszczenie

Celem badań była analiza biometryczna siedmiu odmian lnu oleistego charakteryzujących się zróżnicowaną aktywnością wiązania kadmu z gleby w nasionach pod względem czterech cech ilościowych oraz ocena wpływu zwiększenia zawartości kadmu w glebie na ontogenezę roślin lnu. Analizowane odmiany pochodziły z krajowych zasobów genowych rodzaju Linum. Za pomocą wieloczynnikowej analizy wariancji określono zależności pomiędzy badanymi odmianami, zawar-tością kadmu w glebie i latami prowadzenia doświadczeń. Obserwowano istotny wpływ odmian i analizowanych interakcji na badane cechy ilościowe. Zwiększenie zawartości kadmu w glebie do 4 mg/kg w istotny sposób obniżało ogólną masę roślin odmiany ACMC Duff oraz miało istotny wpływ na długość wiechy badanych odmian. Obliczone współczynniki zmienności wykazały niewielkie zróżnicowanie badanych cech ilościowych, niezależnie od zastosowanego stężenia kadmu w glebie oraz lat prowadzenia doświadczeń. Otrzymane wyniki wskazują, że zwiększenie zawartości kadmu w glebie z 2 do 4 mg/kg może mieć istotnie ujemny wpływ na wzrost i rozwój roślin lnu oleistego.

Introduction

Pollution of natural environment with cadmium poses a high threat for humans and livestock. Cadmium can cause serious damages to internal organs (Czeczot and Skrzycki 2010). The total content of cadmium in soils used in agriculture is usually below 0.5 mg/kg while higher contents are of anthropogenic origin (Bjelkova et al. 2011). Various plant species display different activity of cadmium absorption from soil. Kuboi et al. (1987) have classified agricultural plant species in terms of this trait and have identified a group of low, medium and high activity of cadmium absorption from soil. Flax (Linum usitatissimum L.) is the species characterized by a high binding ability of cadmium from soil and its accumulation in the seeds (Angelova et al. 2004, Wielebski 2012). Therefore the cadmium content in seeds of industrial crops is usually in the range of 0.05–0.2 mg/kg (Bjelkova et al. 2011). The limit for cadmium in flaxseed by Commission Regulation EC 2006 is 0.1 mg/kg (Wielebski 2012). High cadmium content is an important barrier to the use of flaxseed (Rajesh 2013). For years linseed has been known for its healthy properties, mainly in the context of gastrointestinal and cardiovascular diseases. Pressed linseed oil is rich in polyunsaturated omega-3 fatty acids (Wałkowski et al. 1998, Silska and Praczyk 2011). A positive effect of linseed oil was found, inter alia, in anticancer therapy, which now, with consequently increasing number of deaths caused by cancer is of great importance. Numerous studies also consider the health-promoting properties of the lignans, of which the flax seed is a rich source.

Cadmium is not an essential element for the growth and reproduction of plants, and its large quantities can have a negative impact on the plant ontogeny. In this study, the effect of increasing the levels of cadmium in the soil from 2 to 4 mg/kg of soil on 4 economically important, quantitative traits of flax was investigated. The plant height (total length), technical length, panicle length, and total yield were determined. Tests were carried out on seven flax cultivars having diverse ability for accumulating cadmium from soil selected in preliminary experiment.

Materials and methods

The baseline for the analysis were the results of the three-year pot experiments conducted in the greenhouse, at the Institute of Natural Fibres and Medicinal Plants, Experimental Station in Pętkowo (Wielkopolska voivodeship), in 2011–2013.

In each year of the experiment, two concentrations of cadmium in soil were used: 2 and 4 mg of Cd per kilogram of soil. Cadmium was introduced into the soil in the form of cadmium nitrate, supplementing its soils content to the above quantities. The soil came from the farmland of the Experimental Station in Pętkowo (52o

12’40” N 17o15’31” E). It was characterized by a neutral reaction, trace heavy metals content (Cd content 0.1 mg/kg) and belonged to the IVa soil class.

50 cultivars of linseed from Polish collection of Linum genus were tested in terms of their binding ability of cadmium from soil in the first year of the study. Experiment was conducted with three replications. Seeds of tested cultivars were sown in soil containing 2 and 4 mg cadmium per 1 kg of the soil. At the end of the growing season the cadmium content was determined in harvested seed. On this basis, 7 cultivars, characterized by the differential ability of cadmium accumulation (the highest and the lowest amount of cadmium in seeds) were selected for further study. Four selected cultivars were characterized by low cadmium binding ability from soil, while three – by high cadmium binding ability.

Table 1 List of linseed cultivars with different ability of cadmium accumulation from soil

Wykaz odmian lnu oleistego o zróżnicowanej aktywności pobierania kadmu z gleby

No. — Lp. Variety — Odmiana Description — Opis

1 Abby

Cultivars with low ability of cadmium absorption from soil Odmiany o niskiej aktywności wiązania kadmu z gleby

2 Dufferin

3 Lindor

4 La Estanzuela 117

5 Shanxi

Cultivars with high ability of cadmium absorption from soil Odmiany o wysokiej aktywności wiązania kadmu z gleby

6 ACMC Duff

7 Alessandra

The cultivars listed in Table 1 were sown randomly in 2012 and 2013 in 3 replications, in the soil containing 2 mg and 4 mg of cadmium/kg of the soil. 30 seeds from tested cultivars were sown in the pots. The biometric measurements were performed for 20 plants harvested from each object (pot).

The tested cultivars were analyzed for the following quantitative traits: 1) plant height – a distance from the root collar to the top of the plant,

2) technical length – a distance from the root collar to the first branch of the panicle,

3) panicle length – a distance from the first branch of the panicle to the top of the plant,

4) total plant weight (total yield) – the weight of harvested plants together with roots. The results were analyzed statistically. A multivariate and univariate analysis of variance was performed, taking into account the double and triple interactions between the years of the experiment and the used cultivars, as well as the concentration of cadmium in the soil. We also calculated the variability coefficients for the traits analyzed, taking into account both years of experiment and two applied concentrations of cadmium in the soil. Calculations were performed using the statistical package Statistica v. 10.

Results and discussion

Based on multivariate analysis of variance was discarded testing hypothesis about the lack of differences between cultivars, cadmium content in the soil and years. A hypothesis concerning the lack of interactions was also rejected. Lack of significance at the level of 0.001 was found only for the cadmium content (Tab. 2). Due to significant interactions further analyses were performed for each trait independently.

Table 2 Multivariate analysis of variance — Wielowymiarowa analiza wariancji

Source of variability

Źródło zmienności Wartość Value Statystyka F F

Effect df Efekt df Error df Błąd df P Wartość p Year — Rok 0.156 97.11 3 54.00 <0.001 Cultivar — Odmiana 0.044 17.02 18 153.22 <0.001 Year × cultivar Rok × odmiana 0.095 11.01 18 153.22 <0.001 Cadmium content Zawartość kadmu 0.794 4.65 3 54.00 0.005 Year × Cd content

Rok × zawartość kadmu 0.702 7.61 3 54.00 <0.001

Cultivar × Cd content

Odmiana × zawartość kadmu 0.413 3.12 18 153.22 <0.001

Year × cultivar × Cd content

Rok × odmiana × zawartość kadmu 0.435 2.90 18 153.22 <0.001

Univariate analysis of variance showed significant differences (P < 0.001) of examined linseed cultivars in terms of all analyzed quantitative traits (Tab. 3, 4). All the traits were significantly affected by the conditions in the year of the experiment (Tab. 3, 4). Constant model for observations was assumed due to the large diversity of the impact of conditions in the years for traits. The content of cadmium in the soil significantly determined the value of panicle length and total yield, however only the panicle length at a significance level of 0.001 (Tab. 3, 4). For the total weight of plants significant influence of cadmium content in the soil was on the 0.05 level (Tab. 5, 6). Based on the analysis of variance, an interaction between year of the experiment and tested cultivars for all analyzed traits were found (Tab. 3, 4). In the case of plant height and technical length the interaction between the year of the experiment and the applied concentration of cadmium in the soil was found (Tab. 3, 4). For the panicle length the interaction cultivar × concentration of cadmium in the soil (at the level of 0.001), as well as year × cultivar × Cd content (at the level of 0.05) was found (Tab. 3, 4).

Table 3 Analysis of variance for plant height and technical length

Jednowymiarowa analiza wariancji dla wysokości roślin i długości technicznej

Source of variability

Źródło zmienności df

Plant height

Wysokość roślin Długość techniczna Technical length

MS F p MS F p

Year — Rok 1 283.8 22.92 < 0.001 138.3 12.87 < 0.001

Cultivar — Odmiana 6 390.1 31.50 < 0.001 567.8 52.84 < 0.001 Cadmium content in soil

Zawartość kadmu w glebie 1 32.2 2.60 0.112 0.5 0.04 0.837

Year × cultivar

Rok × odmiana 6 142.4 11.50 < 0.001 132.3 12.32 < 0.001

Year × Cd content

Rok × zawartość kadmu 1 140.9 11.38 0.001 203.1 18.89 < 0.001

Cultivar × Cd content

Odmiana × zawartość kadmu 6 26.0 2.10 0.067 19.2 1.79 0.117

Year × cultivar × Cd content

Rok × odmiana × zawartość kadmu 6 32.8 2.65 0.024 27.1 2.52 0.031

Error — Błąd 56 12.4 10.7

Total — Ogółem 83

Table 4 Analysis of variance for panicle length and total yield

Jednowymiarowa analiza wariancji dla długości wiechy i ogólnej masy roślin Source of variability

Źródło zmienności df

Panicle length

Długość wiechy Ogólna masa roślin Total yield

MS F p MS F p

Year — Rok 1 25.85 12.803 < 0.001 365.00 108.797 < 0.001

Cultivar — Odmiana 6 34.04 16.857 < 0.001 37.86 11.284 < 0.001 Cadmium content in soil

Zawartość kadmu w glebie 1 24.97 12.367 < 0.001 25.74 7.673 0.007

Year × cultivar Rok × odmiana

6 9.28 4.596 < 0.001 40.71 12.134 < 0,001 Year × Cd content

Rok × zawartość kadmu

1 5.66 2.802 0.099 0.26 0.078 0.780

Cultivar × Cd content

Odmiana × zawartość kadmu 6 8.94 4.429 < 0.001 6.16 1.835 0.108

Year × cultivar × Cd content

Rok × odmiana × zawartość kadmu 6 7.47 3.701 0.003 5.62 1.674 0.144

Error — Błąd 56 2.02 3.35

Different influence of the years on the investigated traits was observed. In the first year of the experiment, most of the analyzed cultivars exhibited greater total length and technical length at a cadmium concentration in soil of 4 mg/kg. In the second year, most cultivars were characterized by higher values of the total length and technical length at the cadmium concentration in soil of 2 mg/kg. The total weight of the plant and the panicle length were higher in most varieties at soil cadmium concentrations of 2 mg/kg. These results were found in both years of the experiment (Tab. 5, 6). As mentioned, the content of cadmium in the soil significantly determined the value of the total weight of the plant and panicle length (Tab. 5, 6). Significant effect of cadmium content in the soil for total yield

Table 5 Influence of cadmium content in the soil on average values of the 4 quantitative traits for 7 linseed cultivars in the first year of experiment — Wpływ zawartości kadmu w glebie na średnie wartości 4 cech ilościowych dla 7 odmian lnu oleistego w pierwszym roku prowadzenia doświadczenia (Pętkowo, 2012)

Cultivar Odmiana Cadmium content in soil Zawartość kadmu w glebie Plant height Wysokość roślin [cm] Technical length Długość techniczna [cm] Panicle length Długość wiechy [cm] Total yield Ogólna masa roślin [g] Abby 2 mg/kg 64.9 49.0 15.9 21.1 4 mg/kg 67.7 53.2 14.5 20.5 LSD — NIR0,05 nd nd nd nd Shanxi 2 mg/kg 71.7 55.7 15.4 22.9 4 mg/kg 70.7 61.7 9.0 22.7 LSD — NIR0,05 nd nd 5.80 nd Alessandra 2 mg/kg 73.4 61.2 12.2 19.5 4 mg/kg 70.0 62.0 8.0 18.1 LSD — NIR0,05 nd nd 3.66 nd Estanzuela 117 2 mg/kg 69.3 57.9 11.4 22.9 4 mg/kg 72.8 60.6 12.2 21.2 LSD — NIR0,05 nd nd nd nd ACMC Duff 2 mg/kg 75.4 65.1 10.3 25.5 4 mg/kg 82.6 70.7 11.9 21.0 LSD — NIR0,05 nd nd 1.49 4.06 Lindor 2 mg/kg 66.7 57.0 9.7 21.3 4 mg/kg 63.2 53.5 9.7 21.7 LSD — NIR0,05 nd nd nd nd Dufferin 2 mg/kg 64.5 52.4 12.1 14.2 4 mg/kg 67.2 56.8 10.4 14.2 LSD — NIR0,05 nd nd nd nd

of ACMC Duff cultivar in the both years of experiment was observed. Increasing cadmium content in the soil from 2 mg/kg to 4 mg/kg resulted in a significantly lower total yield of ACMC Duff cultivar in both years of experiment (Tab. 5, 6). Significant influence of cadm content in soil on the length of panicle was observed only in the first year of research. ACMC Duff cultivar showed significantly longer panicle at the concentration of cadmium in the soil at the level of 4 mg/kg. Shanxi and Alessandra cultivars had significantly longer panicles at the concentration of cadmium in the soil at the level of 2 mg/kg (Tab. 5).

Table 6 Influence of cadmium content in the soil on average values of the 4 quantitative traits for 7 linseed cultivars in the second year of experiment — Wpływ zawartości kadmu w glebie na średnie wartości 4 cech ilościowych dla 7 odmian lnu oleistego w drugim roku prowadzenia doświadczenia (Pętkowo, 2013)

Cultivar Odmiana Cadmium content in soil Zawartość kadmu w glebie Plant height Wysokość roślin [cm] Technical length Długość techniczna [cm] Panicle length Długość wiechy [cm] Total yield Ogólna masa roślin [g] Abby 2 mg/kg 60.7 44.0 16.7 13.6 4 mg/kg 57.4 43.0 14.4 13.2 LSD — NIR0,05 nd nd nd nd Shanxi 2 mg/kg 72.5 58.5 14.0 17.7 4 mg/kg 65.5 51.7 13.8 16.5 LSD — NIR0,05 nd nd nd nd Alessandra 2 mg/kg 76.7 61 15.7 15.6 4 mg/kg 79.8 65.9 13.9 17.7 LSD — NIR0,05 nd nd nd nd Estanzuela 117 2 mg/kg 75.3 64.6 10.7 15.2 4 mg/kg 75.1 63 12.1 16.0 LSD — NIR0,05 nd nd nd nd ACMC Duff 2 mg/kg 87.6 75.8 11.8 20.7 4 mg/kg 78.6 67.4 11.2 17.3 LSD — NIR0,05 nd nd nd 3.15 Lindor 2 mg/kg 84.7 72.1 12.6 16.9 4 mg/kg 74.9 65 9.9 12.7 LSD — NIR0,05 nd nd nd nd Dufferin 2 mg/kg 71.7 61.9 9.8 17.8 4 mg/kg 74.1 59.3 11.8 17.0 LSD — NIR0,05 nd nd nd nd

Flax cultivars are characterized by an individualized response to cadmium in soil, which is confirmed by many authors (Morgan 1993, Angelova et al. 2004, Bjelkova et al. 2011). The results of previous experiments indicate that the cadmium content in harvested seeds of certain cultivars is slightly lower than the amount of cadmium introduced into the soil (Praczyk et al., unpublished).

Grant et al. (1997) reported a slight impact of elevated levels of cadmium in soil on the ontogeny of many crop species, however, no interaction between cultivars, the cadmium content in soil and years of the experiment was determined. The presented results suggest that negative effect of higher cadmium content in soil (4 mg/kg) on the total yield of flax cultivars can be established, especially in cultivars with high ability of cadmium absorption from soil.

In the present study, also the variability of all analyzed quantitative traits was calculated. The values of the variation coefficients indicate that this variability was low and remained at a similar level, regardless of the concentration of cadmium in soil and year of the experiment (Tab. 7, 8). The highest coefficient of variation in the first year of the study, for both concentrations of cadmium in soil was found for panicle length, while in the second year of the study – for the technical length (Tab. 7, 8).

Many authors mention low variability of important flax traits (Popescu et al. 1999, Sood et al. 2007, Singh et al. 2009). Only slight differences in cultivars occur even between genotypes originating from geographically distant regions, which demonstrates the low variability of the initial materials for breeding cultivars of fibrous and oil flax (Rólski et al. 2001, Praczyk and Silska 2013). The results obtained in this study confirmed the ones reported in the literature in terms of low variability of flax features, indicating at the same time that the cadmium found in soil, even in relatively high amounts (4 mg/kg) has no effect on the variability of features.

Tab Te ste d tr aits v ar ia bilit y fo r m ea n f ro m 7 li ns ee d c ulti va rs in th e f irs t y ea r o f e xp er im en t Zm ie nnoś ć bada ny ch ce ch dl a śr edni ej z 7 odm ian ln u ol ei ste go w pi er ws zy m rok u pr ow adz eni a doś wi adc ze ni a (P ęt ko w o, 20 12) Tr ai t C ech a C d co nte nt in so il 2 mg /k g Zaw ar toś ć kadm u w g le bi e 2 m g/ kg C d co nte nt in so il 4m g/k g Zaw ar toś ć kadm u w g le bi e 4 m g/ kg ra ng e bet w een th e lo w es t a nd h ig he st va lu e — ro zs tę p sta nd ar d d ev ia tio n od ch yl eni e st an da rd ow e va ria bi lity c oe ffic ie nt w sp ół cz yn ni k zm ie nn oś ci [%] ra ng e b et we en th e lo w es t a nd h ig he st va lu e — ro zs tę p sta nd ar d d ev ia tio n od ch yl en ie st an da rd ow e va ria bil ity c oe w sp ół cz yn zm ie nn oś ci Pla nt h eig ht W yso ko ść ro śl in [c m] 10. 9 4. 21 6.1 19. 4 6.1 8.6 Pan icl e l en gt h D ługoś ć w ie chy [c m] 6.2 2. 38 19. 2 6.5 2. 21 20. 4 Tech ni cal len gt h D łu go ść tech ni cz na [c m] 16. 1 5. 33 9.4 17. 5 6. 05 10. 1 To ta l y ie ld O gól na m as a roś lin [g ] 11. 3 3. 56 16. 9 8.5 2. 88 14. 5 Tab Te ste d tr aits v ar ia bilit y fo r m ea n f ro m 7 li ns ee d c ulti va rs in th e seco nd y ear o f e xp er im en t Zm ie nnoś ć bada ny ch ce ch dl a śr edni ej z 7 odm ian ln u ol ei ste go w dr ugi m rok u pr ow adz eni a doś wi adc ze ni a (P ęt ko wo , 2013 ) Tr ai t C ech a C d co nte nt in so il 2 m g/k g Zaw ar toś ć kadm u w g le bi e 2 m g/ kg C d co nte nt in so il 4m g/k g Zaw ar toś ć kadm u w g le bi e 4 m g/ kg ra ng e bet w een th e lo w es t a nd h ig he st va lu e — ro zs tę p sta nd ar d d ev ia tio n od ch yl eni e st an da rd ow e va ria bi lity c oe ffic ie nt w sp ół cz yn ni k zm ie nn oś ci [%] ra ng e bet w een th e lo w es t a nd h ig he st va lu e — ro zs tę p sta nd ar d d ev ia tio n od ch yl en ie st an da rd ow e va ria bil ity c oe w sp ół cz yn zm ie nn oś ci Pla nt h eig ht W yso ko ść ro śl in [c m] 26. 9 8. 89 11. 8 21. 2 7. 94 11. 1 Pan icl e l en gt h D ługoś ć w ie chy [c m] 6.9 2. 55 19. 5 4.5 1. 65 13. 3 Tech ni cal len gt h D łu go ść tech ni cz na [c m] 31. 8 10. 28 16. 4 24. 4 8. 93 15. 0 To ta l y ie ld O gól na m as a roś lin [g] 7.1 2. 28 13. 6 4.5 2.0 12. 7

Conclusions

1. Some variability of all quantitative traits was observed regardless of the concentration of cadmium in soil and year of experiment. The increase in the cadmium content in soil to 4 mg/kg did not cause any changes in the variability of studied quantitative traits.

2. The results of the variance analysis allowed to determine significant differences between flax varieties tested in terms of total yield and panicle length, as well as the expression of the all tested traits in the years of experiment.

3. A significant influence of the concentration of cadmium in soil on the total weight of the plant and panicle length was found. The increase of cadmium content in soil from 2 to 4 mg/kg resulted in a significantly lower total yield of ACMC Duff cultivar in both years of experiment. This suggests the possibility of negative influence of a relatively small amount of cadmium in soil to reduce linseed crop.

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