CS275634B6 - A regulating effect on algal growth and chlorophyll synthesis - Google Patents

Abstract

The solution relates to a composition with regulatory activity on the growth and synthesis of chlorophyll in green algae. The composition consists of a solution of a 2:-R-3-Rlbenzothiazolium salt of the general formula I, where R is hydrogen, methylthio, ethoxycarbonylmethylthio; Rl is hydrogen, methyl, benzyl, allyl, methoxycarbonylmethyl, ethoxycarbonylmethyl, propoxycarbonylmethyl, isopropoxycarbonylmethyl; X" is a bromide, iodide, sulfate, methosulfate group. Compounds of general formula I have an inhibitory effect on the growth and synthesis of chlorophyll of algae at concentrations in the culture medium of 5.3 mg.dm-3 and higher. Benzothiazolium salts in which R1 is alkoxycarbonylmethyl, as well as derivatives in which R is ethoxycarbonylmethylthio, have a stimulatory effect on the growth and synthesis of chlorophyll of algae in the concentration range of 3.2 mg.dm-3 to 0.03 ng.dm-3.

Description

The invention relates to a composition with a regulating effect on the growth and synthesis of green algae chlorophyll, which consists of a solution of a benzothiazolium salt of the general formula I

wherein R is hydrogen, methylthio, ethoxycarbonylmethylthio;

R ¹ is hydrogen, methyl, benzyl, allyl, methoxycarbonylmethyl, ethoxycarbonylmethyl, propoxycarbonylmethyl, isopropoxycarbonylmethyl:

X is a bromide, iodide, sulfate, methosulfate group.

The benzothiazolium salt may be in solution in organic solvents such as dimethyl sulfoxide, methyl alcohol, triethylene glycol, N, N-dimethylformamide, but an aqueous solution may be preferably used. '

Benzotiazoliové salt of the formula I act to inhibit the synthesis of chlorophyll and growth of green algae at concentrations in the culture medium of 5.3 mg.dm ^- ^ and above. At these concentrations, they also cause photodegradation of chlorophyll in isolated plant chloroplasts. Derivatives in which R ¹ is alkoxycarbonylmethyl and also a derivative in which R is alkoxycarbonylmethylthio act to stimulate the growth and synthesis of green algae chlorophyll in a concentration range of 3.2 mg-dm 2 to 0.03 ng.dm 2 ^.

Some benzothiazolium salts are effective as plant growth regulators (Sutoris V., Halgaš 3., Sekerka V .: Czechoslovak Authorization Nos. 225008, 223426, 231212, 231229, 233443; Sutoris V., Gáplovský A., Sekerka V .: Chem.Papers 40 (1) 103, 1986), or they act specifically to increase the sugar content in sugar-producing plants (Sutoris V., Sekerka V., Halgaš 3., Bajči P., Konečný V., Varkonda Š .: Czechoslovak Author's Certificate No. 237706). The benzothiazolium salts were found to have antimicrobial activity against G ⁺ bacteria of the Staphylococcus group (Sutoris V., Foltínová P., Halgaš 3 .: Czech author's certificate No. 237709) as well as antifungal activity (Sutoris V., Halgaš 3., Sekerka V ., Folínová A., Gáplovský A .: Chem.Zvěsti 37, 653-662, 1983).

The inhibitory activity of the compounds of the formula I on algae was tested by stationary cultivation of a synchronous culture of Chlorella vulgaris. The growth of algal cells and the content of synthesized chlorophyll were evaluated. The inhibitory activity of these compounds is expressed by the factor IC50, which represents the concentration of a given benzothiazolium salt in the algal culture medium that causes 50% inhibition of the growth and growth of the algae, respectively. chlorophyll synthesis, with respect to algal control samples. '

Regulatory activity on the growth and synthesis of Chlorella vulgaris chlorophyll was monitored on the following benzothiazolium salt derivatives: 1. 3-methylbenzothiazolium iodide 2. 3-allylbenzothiazolium bromide 3. 3-benzylbenzothiazolium bromide 4. 2-methylthio-3-thiethylmethenzothenzene 3-methylbenzothiazolium iodide 6. benzothiazolium sulfate 7. 3-methoxycarbonylmethylbenzothiazolium bromide 8. 3-ethoxycarbonylmethylbenzothiazolium bromide 9. 3-propoxycarbonylmethylbenzothiazolium bromide 10. 3-isopropoxycarbonylmethylbenzothiazolium 2-benzothiazolium

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The IC 50 values of the compounds of the formula I are given in Tab. 1. For comparison, we note that the inhibitory activity of 2-niercaptobenzothiazole, determined by the same method and expressed by an IC ₅₀ factor, is 68.4 / Uniol. dm ^- · ^{5 for} algal chlorophyll synthesis and 80.4 / Umol'din ^-5 for algal growth. As follows from the values in tab. 1, the inhibitory activity of 3-methylbenzothiazolium iodide and 3-allylbenzothiazolium bromide on algae is higher than that of standard 2-nitrocaptobenzothiazole.

The stimulatory activity of benzothiazole salts on algal chlorophyll growth and synthesis is shown in Tab. 2.

Table 1

Inhibitory activity of the compounds of formula I on algae Chlorella vulgaris

Sample no. R R ¹ X ICjq /umól.dm chlorophyll synthesis rastu rias' 1 H ch ₃ I 37.0 54.0 2 H ch ₂ ch = ch ₂ Br 58.6 65.2 3 H CH-C.H, Loj Br 152 316 4 sch ₃ ch ₃ ch ₃ so ₄ 171 188 5 sch ₃ ch ₃ I 160 182 6 H H hso ₄ 598 776 7 H CH ₂ C00CH ₃ Br 375 398 8 H CH ₂ C00C ₂ H ₅ Br 288 344 9 H CH ₂ C00C ₃ H ₇ Br 261 273 10 H CH ₂ COOC ₃ H ₇ -i Br 293 365 11 sch ₂ coo-c ₂ h ₅ ch ₃ Br 242 249

Table 2

The stimulatory activity of the compounds according to the invention on the growth and synthesis of the chlorophyll of the alga Chlorella vulgaris, expressed as a percentage of the control sample

Sample no. % of control ί θ 'x / Chlorophyll synthesis Rast algae 7 119.4 and 18.5 109.3 and 14.7 8 113.9 and 20.7 103.4 and 7.9 9 123.2 - 25.9 106.5 and 13.4 10 120.1 and 16.1 105.8 and 11.3 11 129.2 and 19.0 114.9 and 12.6

^χ / average of the concentration range 1.10 ^- ^ mol dm ^"3 to 1.10 ^- niól.dnt ^^ ^- ^

CS 275634 B 6

The regulatory activity of the benzothiazolium salts of formula I on algae and plants is demonstrated in more detail by way of examples.

Example 1

The effect of bile ducts according to the invention on the growth of the algae Chlorella vulgaris and the content of synthesized chlorophyll

Chlorella vulgaris algae were grown by stationary cultivation at 25-1 ° C under a light regime of 16 hours light for 8 hours dark, in the culture medium according to Šetlík (Šetlík I .: Annual Report of Algolog. Lab. For 1967, 71-140, Třeboň 1968). A solution of the study compound in distilled water was added to the nutrient medium in such a concentration as to obtain a series of culture flasks with a graded concentration of substance and a graded biological effect. The flasks were inoculated with the exact volume of a homogeneous algae suspension. After culturing for 7 days, the volume of the culture medium was made up to the original volume with distilled water (water evaporation compensation) and the absorbance at 660 nm (proportional to the possibility of algal cells) and chlorophyll content after extraction with N, N'-dimethylformamide from centrifuged cells were measured in the cell suspension. rias. The total chlorophyll content of algae in the culture medium was evaluated according to the equation:

Chl (mg.dm ” ³ ) = 17.9 A ^^^ + 8.08 A ^^ where A is the absorbance of the sample at 647 resp. Swing 664.5 nm in 1 cm (Inskeep WP, Bloom PR: Plant Physiol. 77/2 /, 483-5, 1985).

The obtained values of these parameters were compared with the values of the parameters of the simultaneously tested control samples (evaluated as% of the control). Based on these values, the concentrations of the studied compounds at which 50% inhibition was achieved (IC 50) were calculated by extrapolation. In tab. Figure 3 shows the regulatory activity of 3-isopropoxycarbonylmethylbenzothiazolium bromide on algal chlorophyll growth and synthesis as a function of applied concentration.

The values of the concentration of the studied compound at which there is a 50% inhibition of algal growth resp. chlorophyll syntheses are determined by extrapolation from the dependence of the inhibition values found on the logarithm of the concentration of study compound applied.

Table 3

Regulatory activity of β-isopropoxycarbonylmethylbenzothiazolium bromide on the growth and synthesis of chlorophyll algae Chlorella vulgaris

Concentration % control - O ' Chlorophyll content Rast algae i.io ” ³ 0 8.2 and 0.1 5.10 ” ⁴ 0 22.6 and 0.4 3.10 ” ⁴ 47.6 and 3.5 66.0 - 0.4 2.10 ” ⁴ 53.5 and 3.6 72.0 and 0.1 1.10 ' ⁴ 96.0 and 2.6 93, B - 4.7 1.10 ^-5 117.7 - 20.3 102.3 - 12.5 1. ID ” ⁷ 128.7 and 12.0 108.3 and 11.1 1.10 ” ⁹ 122.5 and 17.8 109.5 - 11.3 1.10 ^-11 130.5 - 2.1 118.6 and 1.8 1.10 ” ¹³ 110.1 and 14.7 103.2 and 8.2

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Example 2

Influence of benzothiazolium salts on chlorophyll photodegradation in chloroplastic preparations

To a preparation of chloroplasts prepared from spinach using TRIS buffer (2.10 mol.dm; pH 7.0) and sucrose (0.4 mol.dm), an aqueous solution of benzothiazolium salts was added, and the samples were irradiated with a neon lamp for 4 hours after homogenization. The original chlorophyll content in the non-irradiated samples was 23 mg.dm ^-3 . Then, the irradiated samples were diluted with acetone in a ratio of 4: 1 and the decrease in chlorophyll absorbance at a wavelength of 664.5 nm with respect to the absorbance of the non-irradiated control sample was determined spectrophotometrically. At the tested concentrations of benzothiazolium salts (1.10 ^- to 5.10 niol.dm) a significant increase in chlorophyll photodegradation was observed. An analogous increase in the photodegradation of the growth of lineage pigments (chlorophyll, carotene) due to the higher concentrations of benzothiazolium salts was also determined in solutions of N, N'-dimethylformamide. Table 4 shows the values of the effect of chlorophyll absorbance at a wavelength of 664.5 nm in a chloroplast preparation with 3-ethoxycarbonylmethylbenzothiazolium bromide after irradiation of the samples for 4 hours.

Table 4

Influence of chlorophyll absorbance at a wavelength of 664.5 nm in a preparation of 3-ethoxycarbonylmethylbenzothiazolium bromidone spinach chloroplasts after 4 hours of irradiation of the samples (the original chlorophyll content in the samples was 24 mg.dm)

Molm.dm concentration ”3 % Original chlorophyll absorbance 0 (check) 63.3 1.10 ' ² 15.8 5.10 ³ 19.7 1.10 ³ 26.0 5.10 ⁴ 30.7 2.10 ^-4 40.9 1.10 ' ⁴ 43.5 5.10 ^-5 49.2 1.10 “ ⁵ 63.2

Example 3

Influence of benzothiazolium salts on fresh mass growth and chlorophyll content in the aboveground part of maize

To test the increase in fresh mass as well as the content of synthesized chlorophyll in the aboveground part of corn, corn seeds were pre-germinated in the dark in rolls of rolled filter paper moistened with distilled water for 3 days. The key ring placed in hydroponics comprising aqueous solutions of salts benzotiazolivoých concentrations of 1.10 ^-3 to 1.10 ^- ^ ^-5 mol.dm and placed in a culture box with a light regime of 14 hours light, 10 hours dark. After 6 days of culturing at room temperature, the weight of the green part of the germination and the chlorophyll content were determined.

At the concentrations of benzothiazolium salts tested, a stimulating effect was observed both on the increase in fresh mass and on the content of synthesized chlorophyll in the aboveground part of maize. Table 5 shows the stimulatory effects of 3-non-methoxycarbonylmethylbenzothiazolium bromide and 3-methylbenzothiazolium iodide on fresh mass growth and the content of synthesized chlorophyll in the aboveground part of maize.

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Table 5

Stimulatory activity of 3-methoxycarbonylmethylbenzothiazolium bromide and 3-methylbenzothiazolium iodide on fresh mass growth and synthesized chlorophyll content in the aboveground part of maize

Sample no. Mole concentration. dm % Controls - Ι.5.θ Chlorophyll content Weight stalks 0 (check) 100.0 and 5.1 100.0 + 5.4 1.10 ⁵ 109.2 and 5.8 98.6 + 3.4 7 1.10 ' ⁶ 110.7 and 0.7 113.0 + 4.2 1.10 ⁸ 111.2 and 6.5 107.5 + 2.2 i.io- ¹⁰ 102.3 and 1.1 111.7 + 1.3 1.10 ' ⁵ · 115.5 and 6.4 108.9 + B, 2 1 1.10 ^-8 117.0 and 5.6 115.1 + 2.6 1.10 “ ¹¹ 120.4 - 3.8 133.9 + 2.5

PATENT CLAIMS

Claims (3)

Table 5 Stimulation efficiency of 3-methoxycarbonylmethylbenzothiazolium bromide and 3-methylbenzothiazolium iodide for the increase in fresh mass and the content of synthesized chlorophyll in the maize sample Sample No. Concentration mol. dm% Controls - IS-θ Chlorophyll content Stalk Weight 0 (control) 100.0 ,1 5.1 100.0 ,4 5.4 1.10 5 5 109.2 ,8 5.8 98.6 i 3.4 7 1.10 ' 6 110.7 0.7 113.0 i 4.2 1.10 "8 111.2 i 6.5 107.5 i 2.2 i.io-10 102.3 i 1.1 111.7 - 1.3 1.10'5 115.5 I 6.4 108.9 i 0.2 1 1.10-8 117.0 I 5.6 115.1 I 2.6 1.10 ”11 120.4 - 3.8 133.9 i 2 , 5 PATENT CLAIMS 1. An algal growth regulating and chlorophyll synthesis composition comprising as active ingredient 2-R-3-R1 benzothiazolium salt of formula I wherein R is hydrogen, methylthio, ethoxycarbonylmethylthio; R 1 is hydrogen, methyl, benzyl, allyl, methoxycarbonylmethyl, ethoxycarbonylmethyl, propoxycarbnonylmethyl, isopropoxycarbonylmethyl; X is a bromide, iodide, sulfate, methosulfate group. 2. A composition according to item 1 with antiallergic activity, characterized in that the concentration of the active compound is 5.3 mg.dm-3 or higher. 3. A composition according to claim 1, wherein the growth stimulatory effect on algal growth and chlorophenyl synthesis is characterized in that the concentration of the active ingredient is between 3.2 mg.dm -3 and 0.03 ng.dm where R is ethoxycarbonylmethylthio.