Ice‐‐bacteria as antagonists in biological frost protection

Investigated was the efficacy of 4 ice nucleation‐inactive (ice^‐) bacterial strains (A506, GSPB 1147, 1181, 2357) at 4 ice nucleation active (ice⁺) strains (553, 554, GSPB 1139, 2035) on the surface of 4 culture crops (corn, tomato, bush‐ and field bean). Examinated was the reduction of frost damage at the used culture crops by inoculation with ice⁺ and ice^‐ strains at 4 different variants (A‐D) under laboratory conditions. The ice nucleation activity was determined by the tube‐freezing‐assay. An effective reduction of ice⁺ bacteria was possible, when plant surfaces were preinoculated (3days) with ice^‐bacteria before ice⁺ bacterial strains colonized the surfaces of plants. A statistical comparison of mean values of obtained results showed, the ice crystallization (INT) by inoculation started at ‐3°C and in mean (MNT) below ‐5°C independent of the used ice⁺ and ice^‐ strains. Obtained differences were not significant. As untreated as colonized plants with antagonists started to freeze at ‐5 and ‐6°C and in average at ‐7°C. However, the preinoculation resulted in 1.38 K differences for the temperatures, at which 50% of leaves were frozen (PROZ 50) in favour of preinoculated plants. This difference in freeze temperatures by preinoculation with ice^‐ bacteria was discussed as a possible method of frost protection.     

Plant protection and rhizosphere colonization of barley by seed inoculated herbicide degrading Burkholderia (Pseudomonas) cepacia DBO1(pRO101) in 2,4-D contaminated soil

The 2,4-dichlorophenoxyacetic acid (2,4-D) degrading bacterium, Burkholderia cepacia (formerly Pseudomonas cepacia) DBO1(pRO101) was coated on non-sterile barley (Hordeum vulgare) seeds, which were planted in two non-sterile soils amended with varying amounts of 2,4-D herbicide. In the presence of 10 or 100 mg 2,4-D per kg soil B. cepacia DBO1(pRO101) readily colonized the root at densities up to 10⁷ CFU per cm root. In soil without 2,4-D the bacterium showed weak root colonization. The seeds coated with B. cepacia DBO1(pRO101) were able to germinate and grow in soils containing 10 or 100 mg kg^–1 2,4-D, while non-coated seeds either did not germinate or quickly withered after germination. The results suggest that colonization of the plant roots by the herbicide-degrading B. cepacia DBO1(pRO101) can protect the plant by degradation of the herbicide in the rhizosphere soil. The study shows that the ability to degrade certain pesticides should be considered, when searching for potential plant growth-promoting rhizobacteria. The role of root colonization by xenobiotic degrading bacteria is further discussed in relation to bioremediation of contaminated soils.     

Characterization of the cultivable microbiota of sprouts and their potential for application as protective cultures

The microbiota of ten seeds and ready-to-eat sprouts produced thereof was characterized by bacteriological culture and denaturing gradient gel electrophoresis (DGGE) of amplified DNA fragments of the 16S rRNA gene. The predominant bacterial biota of hydroponically grown sprouts mainly consisted of enterobacteria, pseudomonades and lactic acid bacteria (LAB). For adzuki, alfalfa, mung bean, radish, sesame and wheat, the ratio of these bacterial groups changed strongly in the course of germination, whereas for broccoli, red cabbage, rye and green pea the ratio remained unchanged. Within the pseudomonades, Pseudomonas gesardii and Pseudomonas putida have been isolated and strains of the potentially pathogenic species Enterobacter cancerogenes and Pantoea agglomerans were found as part of the main microbiota on hydroponically grown sprouts. In addition to the microbiota of the whole seedlings, the microbiota of root, hypocotyl and seed leafs were examined for alfalfa, radish and mung bean sprouts. The highest and lowest total counts for aerobic bacteria were found on seed leafs and hypocotyls, respectively. On the other hand, the highest numbers for LAB on sprouts were found on the hypocotyl. When sprouting occurred under the agricultural conditions, e.g. in soil, the dominating microbiota changed from enterobacteria to pseudomonades for mung beans and alfalfa sprouts. No pathogenic enterobacteria have been isolated from these sprout types. Within the pseudomonades group, Pseudomonas jessenii and Pseudomonas brassicacearum were found as dominating species on all seedling parts from soil samples. In practical experiments, a strain of P. jessenii was found to exhibit a potential for use as protective culture, as it suppresses the growth of pathogenic enterobacteria on ready-to-eat sprouts.