PAENIBACILLUS POLYMYXA КВ — PRODUCER OF BIOLOGICALLY ACTIVE SUBSTANCES

L. Tokmakova, L. Shevchenko, A. Trepach
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Abstract

Objective. To investigate the ability of Paenibacillus polymyxa KB to produce biologically active substances that stimulate the growth and development of plants. Methods. Qualitative and quantitative determination of auxins, abscisic acid (ABA) and cytokinins in the culture fluid (CF) of P. polymyxa KB was carried out by high-performance liquid chromatography (HPLC) using Agilent 1200 liquid chromatograph; gibberellins were measured by high-performance liquid chromatography (HPLC/MS) using Agilent 1200 liquid chromatograph (Agilent Technologies, USA) and Agilent G1956B mass spectral detector. The content of vitamins in P. polymyxa KB was determined using Waters liquid chromatograph with a tqs-micro mass-selective detector. The content of amino acids in CF was studied by ion exchange chromatography on a Sycam amino acid analyser. The ability of P. polymyxa KB to synthesize exopolysaccharides and enzymes was determined using classical microbiological and biochemical research methods. Antifungal properties of the bacterium were studied by the dual culture method. Statistical calculations were carried out using analysis-of-variance method. Results. P. polymyxa KB synthesises phytohormonal compounds: auxins — up to 81.15 μg/g of dry biomass, abscisic acid — up to 9.84 μg/g; gibberellic acids (GA3 — within 2.48 μg/g and GA4 — 1.96 μg/g of dry biomass); cytokinins (zeatin — up to 16.42 μg/g, kinetin — up to 6.78 μg/g, isopentenyl-adenine — up to 9.03 μg/g of dry biomass), as well as vitamins: thiamine (B1) — within 0.93 mg/dm3 CF, riboflavin (B2) — 0.48 mg/dm3, pyridoxine (B6) — 0.27 mg/dm3, folic acid (B9) — 0.18 mg/dm3 CF. P. polymyxa KB shows phosphatase, amylolytic, proteolytic, pectolytic activity, produces amino acids: histidine — within 300.4 mg/dm3, alanine — 236.3 mg/dm3, proline — 164.2 mg/dm3, valine — 157.8 mg/dm3, isoleucine — 148.4 mg/dm3, threonine — 126.2 mg/dm3, cystine — 124.0 mg/dm3 CF and others. The bacterium synthesizes exopolysaccharides amylopectin and levan. P. polymyxa KB shows antagonistic activity against phytopathogenic micromycetes from the genera Fusarium, Alternaria, Nigrospora — pathogens of agricultural crops. Conclusion. P. polymyxa KB synthesizes a significant amount of physiologically active substances that can play an important role in the processes of plant organogenesis.
多粘类芽孢杆菌КВ -生物活性物质的生产者
目标。研究多粘类芽孢杆菌KB产生刺激植物生长发育的生物活性物质的能力。方法。采用高效液相色谱法,采用Agilent 1200型液相色谱仪,对多粘菌培养液(CF)中生长素、脱落酸(ABA)和细胞分裂素进行定性和定量测定;采用高效液相色谱法(HPLC/MS)测定赤霉素含量,采用Agilent 1200型液相色谱仪(Agilent Technologies, USA)和Agilent G1956B质谱检测器。采用Waters液相色谱- tqs-微量质量选择检测器对多粘草KB中维生素的含量进行了测定。采用离子交换色谱法,在Sycam氨基酸分析仪上研究了CF中氨基酸的含量。采用经典的微生物学和生化研究方法测定了多粘多糖KB合成外多糖和酶的能力。采用双重培养法研究了该菌的抑菌性能。采用方差分析方法进行统计计算。结果。P. polymyxa KB合成植物激素化合物:生长素-高达81.15 μg/g干生物量,脱落酸-高达9.84 μg/g;赤霉素酸(GA3 - 2.48 μg/g和GA4 - 1.96 μg/g);细胞分裂素(玉米素-高达16.42 μg/g,动素-高达6.78 μg/g,异戊基腺嘌呤-高达9.03 μg/g),以及维生素:硫胺素(B1) -在0.93 mg/dm3 CF内,核黄素(B2) - 0.48 mg/dm3,吡哆醇(B6) - 0.27 mg/dm3,叶酸(B9) - 0.18 mg/dm3 CF。P. polymyxa KB具有磷酸酶,淀粉酶,蛋白水解,肽水解活性,产生氨基酸。组氨酸- 300.4 mg/dm3,丙氨酸- 236.3 mg/dm3,脯氨酸- 164.2 mg/dm3,缬氨酸- 157.8 mg/dm3,异亮氨酸- 148.4 mg/dm3,苏氨酸- 126.2 mg/dm3,胱氨酸- 124.0 mg/dm3 CF等。这种细菌合成外多糖支链淀粉和左旋酵素。P. polymyxa KB对农作物病原菌镰刀菌属、Alternaria属、Nigrospora属的植物致病菌有拮抗作用。结论。多粘草KB合成大量的生理活性物质,在植物器官发生过程中发挥重要作用。
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