Real-time PCR investigation on the expression of sboA and ituD genes in Bacillus spp

Aims: To investigate the expression of sboA and ituD genes among strains of Bacillus spp. at different pH and temperature. Methods and Results: Different Bacillus strains from the Amazon basin and Bacillus subtilis ATCC 19659 were investigated for the production of subtilosin A and iturin A by qRT‐PCR, analysing sboA and ituD gene expression under different culture conditions. Amazonian strains presented a general gene expression level lower than B. subtilis ATCC 19659 for sboA. In contrast, when analysing the expression of ituD gene, the strains from the Amazon, particularly P40 and P45B, exhibited higher levels of expression. Changes in pH (6 and 8) and temperature (37 and 42°C) caused a decrease in sboA expression, but increased ituD expression among strains from Amazonian environment. Conclusions: Temperature and pH have an important influence on the expression of genes sboA (subtilosin A) and ituD (iturin A) among Bacillus spp. The strains P40 and P45B can be useful for the production of antimicrobial peptide iturin A. Significance and Impact of the Study: Monitoring the expression of essential biosynthetic genes by qRT‐PCR is a valuable tool for optimization of the production of antimicrobial peptides.     

Adsorption and degradation of zearalenone by bacillus strains

Two Bacillus strains; Bacillus subtilis 168 and Bacillus natto CICC 24640 separately adsorbed and degraded zearalenone in liquid media, in vitro. Viable, autoclaved (121°C, 20 min) and acid-treated cells of both strains separately bound more than 55% of zearalenone (ZEN, 20 μg/L) after 30 min and 1-h incubation at 37°C under aerobic conditions, and the amount of ZEN adsorbed was dependent on initial cell volume. In addition, ZEN was degraded by the culture extract of both strains. Degradation by B. subtilis 168 and B. natto CICC 24640 culture extract after 24-h aerobic incubation at 30°C was 81% and 100%, respectively. B. natto CICC 24640 culture extract comprehensively degraded ZEN and, for both strains, no oestrogenic ZEN analogues were present. ZEN degradation was accompanied by carbondioxide emission indicating a decarboxylation reaction. ZEN degradation by the salient B. natto CICC 24640 culture extract varied with initial ZEN concentration, incubation time, temperature and pH. Degradation was enhanced by Mn²⁺, Zn²⁺, Ca²⁺ and Mg²⁺ but impeded by Hg²⁺, Cu²⁺, Pb²⁺, ethylenediaminetetraacetic acid and 1,10-phenanthroline. The degradation reaction is associated with a metalloproteinase of molar mass in the range 31–43 kDa. Overall, the two generally recognised as safe Bacillus strains can, potentially, be utilised for detoxification of zearalenone in food.     

In vitro probiotic and safety attributes of Bacillus spp. isolated from beebread,honey samples and digestive tract of honeybees Apis mellifera

Bacillus species isolated from honeybee Apis mellifera gut, honey and bee bread samples were characterized for their in vitro probiotic and safety attributes. Alpha and γ haemolytic cultures were tested for their antibiotic resistance, antibacterial spectrum, acid and bile tolerance, adhesion ability (auto-aggregation, co-aggregation and hydrophobicity) and phenol tolerance. Safety criteria included evaluation of virulence genes and cytotoxicity percentages. Bacillus isolates inhibited both Gram-positive and Gram-negative pathogens including Staphylococcus aureus, Pseudomonas aeruginosa, Enterococcus faecalis and Streptococcus mutans, while none could inhibit Listeria monocytogenes. Among the isolates, Bacillus subtilis ZH05, ZB03 and ZG025 showed resistance to most of the tested antibiotics and were considered unsafe. B. subtilis (4) and B. licheniformis (1) tolerated acidic pH and bile conditions, never the less were more tolerant in simulated intestinal conditions vis-a-vis gastric conditions. In 0·5% phenol concentrations, B. licheniformis ZH02 showed highest growth, while, B. subtilis ZG029 demonstrated highest auto-aggregation (65 ± 4·6) and hydrophobicity (23 ± 3·6) percentages (P < 0·05). The isolates lacked virulence genes (hblA, hblC, hblD, nhe, cytK and ces), and their cytotoxic percentage on Caco-2 cell lines was ˂15%. Overall, honeybees appear to be a good source of Bacillus species exhibiting typical in vitro probiotic properties, which could be of commercial interest.     

Characteristics and antimicrobial activity of <Emphasis Type="Italic">Bacillus subtilis</Emphasis> strains isolated from soil

Antagonistic Bacillus strains were isolated from soil and analyzed for the purpose of determining whether they could be used as natural biological agents. Primary in vitro screening for antagonism of the isolates was performed against five phytopathogenic mould fungi. Strains TS 01 and ZR 02 exhibited the most pronounced inhibitory effects. They were identified as Bacillus subtilis on the basis of their morphological, cultural and physiology-biochemical properties as well as their hierarchical cluster analysis conducted by means of computer program SPSS. The antimicrobial activity of the strains from cultural medium and sterile filtrate were determined in vitro against a great number of predominantly phytopathogenic fungi and bacteria. TS 01 and ZR 02 strains exhibited very broad and at the same time degree varying antibiotic spectra of activities against both Gram-positive and Gram-negative microorganisms. Many of them were tested against sensitivity to the antimicrobial action of B. subtilis for the very first time. B. subtilis TS 01 and ZR 02 showed highest antifungal activity (sterile zone in diameter over 37 mm) against Alternaria solani, Botrytis cinerea, Monilia linhartiana 869, Phytophthora cryptogea 759/1 and Rhizoctonia sp. The most sensitive bacterial species were found to be Pseudomonas syringae pv. tomato Ro and Xanthomonas campestris with sterile zones 48.0 and 50.0 mm in diameter, respectively. The latter draws a conclusion that the isolated and identified Bacillus subtilis strains are promising natural biocontrol agents and should be further studied and tested for control of numerous plant diseases.     

Selection of Bacillus species for targeted in situ release of prebiotic galacto-rhamnogalacturonan from potato pulp in piglets

We have previously shown that galacto-rhamnogalacturonan fibers can be enzymatically extracted from potato pulp and that these fibers have potential for exerting a prebiotic effect in piglets. The spore-forming Bacillus species are widely used as probiotics in feed supplements for pigs. In this study, we evaluated the option for further functionalizing Bacillus feed supplements by selecting strains possessing the enzymes required for extraction of the potentially prebiotic fibers. We established that it would require production and secretion of pectin lyase and/or polygalacturonase but no or limited secretion of galactanase and β-galactosidase. By screening a library of 158 Bacillus species isolated from feces and soil, we demonstrated that especially strains of Bacillus amyloliquefaciens, Bacillus subtilis, and Bacillus mojavensis have the necessary enzyme profile and thus the capability to degrade polygalacturonan. Using an in vitro porcine gastrointestinal model system, we revealed that specifically strains of B. mojavensis were able to efficiently release galacto-rhamnogalacturonan from potato pulp under simulated gastrointestinal conditions. The work thus demonstrated the feasibility of producing prebiotic fibers via a feed containing Bacillus spores and potato pulp and identified candidates for future in vivo evaluation in piglets.

     

Rapid Detection and Characterization of Surfactin-Producing <Emphasis Type="Italic">Bacillus subtilis</Emphasis> and Closely Related Species Based on PCR

The surfactin production genetic locus (sfp) is responsible for the ability of Bacillus subtilis to produce the lipopeptide biosurfactant, surfactin. This report demonstrates the utility of using PCR of the sfp gene as a means of identifying Bacillus species that produce surfactin. We carried out a hemolysis zone assay, quantitative HPLC and NMR in parallel to ensure that the PCR provided correct results. PCR analyses were performed for the sfp gene on 15 standard strains and 20 field-collected Bacillus spp. isolates native to Taiwan. Among the 15 standard strains, surfactin was produced by seven strains of B. subtilis and two closely related species, B. amyloliquefaciens B128 and B. circulans ATCC 4513. Of the 20 field-collected Bacillus spp. isolates; 16 strains yielded surfactin- positive results with PCR and HPLC. A good correlation was observed. Within the 16 field isolates, B. amyloliquefaciens S13 (452.5 mg/L) and B. subtilis S15 (125.6 mg/L) had high productivity of surfactin. The technique is valuable for finding out potential good yields of surfactin-producing strains. The PCR method we used could also be used to find different species or genera containing homologous genes. This is the first report of the detection of surfactin production by B. amyloliquefaciens and B. circulans based on PCR screening.     

Effect of probiotic Bacillus subtilis Ch9 for grass carp,Ctenopharyngodon idella (Valenciennes, 1844), on growth performance,digestive enzyme activities and intestinal microflora

In the present study, Bacillus subtilis Ch9 was evaluated as a probiotic in grass carp, Ctenopharyngodon idella (Valenciennes, 1844). For 56 days the grass carp (50 ± 2.5 g) were given a feed containing B. subtilis Ch9 in three concentrations: 1.0 × 10⁹ (T₁), 3.0 × 10⁹ (T₂) and 5.0 × 10⁹ (T₃) CFU kg^?1 feed in triplicate treatments. The control group (T₀) was given feed without B. subtilis Ch9 for the same period. Determined were the specific growth rate (SGR), feed conversion ratio (FCR), and digestive enzyme activities in the intestine and hepatopancreas as well as the intestinal microflora. After 56 days, fish receiving the diets supplemented with B. subtilis Ch9 showed significantly higher SGR and lower FCR (P < 0.05) than those fed the control diet. There was no significant different in SGR and FCR among T₁, T₂ and T₃ nor was the survival rate affected (P > 0.05) by the dietary treatments. From days 14 to 56 of the experiment, higher protease, amylase and lipase activities in the foregut, midgut hindgut and hepatopancreas were observed in T₁, T₂ and T₃ (P < 0.05) compared with the control over a short‐term (14–28 days). Enzyme activity did not increase after long‐term feeding with B. subtilis Ch9 (56 days), but was still higher than that of control fish (P < 0.05). Fish fed the probiotic had an increase in trend of total aerobic and facultative anaerobic bacterial quantity (P > 0.05), but the ratio of Bacillus was significantly higher (P < 0.05) than in control fish. The total anaerobic bacterial quantity, Bifidobacterium and Lactobacillus were significantly higher (P < 0.05) in fish fed B. subtilis Ch9 compared with fish fed control feed. In conclusion, an optimum dose of B. subtilis Ch9 could modulate intestinal microflora, induce digestive enzyme activity and potentially promote the digestion and absorption of nutrients, as well as improve the growth performance of grass carp significantly.     

Phylogeny in Aid of the Present and Novel Microbial Lineages: Diversity in Bacillus

Bacillus represents microbes of high economic, medical and biodefense importance. Bacillus strain identification based on 16S rRNA sequence analyses is invariably limited to species level. Secondly, certain discrepancies exist in the segregation of Bacillus subtilis strains. In the RDP/NCBI databases, out of a total of 2611 individual 16S rDNA sequences belonging to the 175 different species of the genus Bacillus, only 1586 have been identified up to species level. 16S rRNA sequences of Bacillus anthracis (153 strains), B. cereus (211 strains), B. thuringiensis (108 strains), B. subtilis (271 strains), B. licheniformis (131 strains), B. pumilus (83 strains), B. megaterium (47 strains), B. sphaericus (42 strains), B. clausii (39 strains) and B. halodurans (36 strains) were considered for generating species-specific framework and probes as tools for their rapid identification. Phylogenetic segregation of 1121, 16S rDNA sequences of 10 different Bacillus species in to 89 clusters enabled us to develop a phylogenetic frame work of 34 representative sequences. Using this phylogenetic framework, 305 out of 1025, 16S rDNA sequences presently classified as Bacillus sp. could be identified up to species level. This identification was supported by 20 to 30 nucleotides long signature sequences and in silico restriction enzyme analysis specific to the 10 Bacillus species. This integrated approach resulted in identifying around 30% of Bacillus sp. up to species level and revealed that B. subtilis strains can be segregated into two phylogenetically distinct groups, such that one of them may be renamed.     

Detection of ferulic acid esterase production by Bacillus spp. and lactobacilli

The production of feruloyl esterase activity by Bacillus spp. and lactobacilli can be detected in an agar-plate assay. The assay involves the substitution of the main carbon source in specific agar with ethyl ferulate. A number of Bacillus spp., predominantly B. subtilis strains, were found to exhibit feruloyl esterase activity by this method. Of the examined lactobacilli, Lb. fermentum (NCFB 1751) showed the highest level of ferulic acid esterase activity. The enzyme was released from harvested cells by sonication and showed pH and temperature optima of 6.5 and 30 °C respectively. Received: 2 February 1998 / Received revision: 20 April 1998 / Accepted: 27 April 1998     

Survival of Plasmid-Containing Bacillus subtilis Released into Mushroom Compost

Abstract The survival of a plasmid-containing Bacillus subtilis released into mushroom compost was investigated. The indigenous Bacillus population of mushroom compost exhibited an antibiotic-resistance profile that was distinguished by almost complete absence of chloramphenicol resistance. Bacillus subtilis containing the chloramphenicol-resistance plasmid pC194 was released into mushroom compost microcosms and populations were monitored at different incubation temperatures. The organism colonized both sterile and untreated compost at 37°C, and to a lesser extent at 50°C, but was eliminated after 30 d at 65°C. Although sporulation of the B. subtilis population occurred within compost, the population was maintained for up to 13 weeks at 50°C, largely as vegetative cells. Experiments in which the B. subtilis host strain, without plasmid, was released demonstrated that plasmid carriage had no effect on the ability of the bacterium to colonize and survive in compost. Furthermore, the size and composition of the indigenous bacterial population was unaffected by the presence of the introduced B. subtilis strain. Virtually no loss of plasmid pC194 from the B. subtilis population in compost was observed, and experiments at low growth rates in chemostats confirmed the stability of this host/vector system in the absence of positive selection pressure. Received: 9 July 1997; Accepted: 20 October 1997     

Construction of a new food-grade expression system for <Emphasis Type="Italic">Bacillus subtilis</Emphasis> based on theta replication plasmids and auxotrophic complementation

A new food-grade expression system was constructed for Bacillus subtilis based on replicative food-grade expression plasmids and auxotrophic complementation. The food-grade B. subtilis host FG01 was created by knockout of the dal locus from the chromosome of B. subtilis 168. Two food-grade expression plasmids pXFGT03 and pXFGT05 were constructed by combining a novel theta-type Bacillus replicon with the B. subtilis endogenous gene dal and P43 promoter; while pXFGT05 was derived from pXFGT03 by deletion of two open reading frames (ORFs) from the original replicon. Upon transformation of FG01 with pXFGT03 or pXFGT05, the host phenotype was complemented on Luria–Bertani agar plates by the plasmid-coded dal gene, which served as a food-grade selection marker for recombinants. Results showed that deletion of the two ORFs had no impact on plasmid replication. A reporter gene bgaB was cloned into pXFGT03 and pXFGT05, respectively, under control of the P43 promoter, and it was successfully expressed in this food-grade expression system. Segregational stabilities of two recombinant plasmids were investigated, and they were fully stable.     

Alkaline protease from <Emphasis Type="Italic">Bacillus sp.</Emphasis> isolated from coffee bean grown on cheese whey

Two strains of Bacillus, one from a culture collection (B. subtilis ATCC 6633) and a wild type (Bacillus sp. UFLA 817CF) isolated during coffee fermentation in the south of Minas Gerais, Brazil, were evaluated in relation to secretion of alkaline proteases. The strains were grown on nutrient broth, nutrient broth with sodium caseinate and nutrient broth with three different concentrations of cheese whey powder for 72 h. Samples were collected at 24-h intervals to evaluate the proteolytic activity, protein content and cell population. Maximum protease activity was observed after 24-h growth for both the microorganisms, a period that coincided with the end of the exponential phase. The specific activity values were, respectively, 839.8 U/mg for B. subtilis ATCC 6633 and 975.9 U/mg for Bacillus sp. UFLA 817CF. The 60% saturation presented the best results for specific protease activity in all the growth culture media tested with B. sp. UFLA 817CF. Bacillus sp. UFLA 817CF showed highest enzymatic activity at pH 9.0 and 40°C in the three culture media tested. The protease obtained from culture of the wild Bacillus strain presented stability at pH 7.0 and considerable heat stability at 40°C and 50°C, and could be an alternative for the industry to utilize cheese whey to produce proteolytic enzymes.     

Antifungal Effects of Bacilysin and Fengymycin from Bacillus subtilis F-29-3 A Comparison with Activities of Other Bacillus Antibiotics

Of the two antifungal antibiotics produced by Bacillus subtilis F-29-3, the dipeptide compound bacilysin inhibits yeasts (and bacteria), whereas the formerly unknown fengymycin, a complex of closely related lipopeptide components, shows antibiotic activity against filamentous fungi. Bacilysin production, formerly known for a few strains only, could be demonstrated for all 12 wild-type cultures of Bacillus subtilis tested during this study. The antibiotic also occurs in some strains of three other Bacillus species considered as closely realted to B. subtilis. Members of the lipopeptide class of antifungal Bacillus metabolites were formed by 8 of 12 Bacillus subtilis-isolates and several other Bacillus strains. The antibiotics of F-29-3 were compared with antifungal metabolites of other Bacillus isolates using TLC, agar-diffusion techniques and tests demonstrating the capacity of six lipopeptide and peptide preparations to protect rice seedlings from phytomycosis due to Rhizoctonia solani. Fengymycin proved to be different from the other compounds tested. It was less toxic to the test plants and protected them better from Rhizoctonia disease than the other antibiotics of the study did.     

Cold-adapted Bacilli isolated from the Qinghai–Tibetan Plateau are able to promote plant growth in extreme environments

Nearly 1400 Bacillus strains growing in the plant rhizosphere were sampled from different sites on the Qinghai–Tibetan Plateau. Forty-five of the isolates, selected due to their biocontrol activity, were genome-sequenced and their taxonomic identification revealed that they were representatives of the Bacillus subtilis species complex (20) and the Bacillus cereus group (9). Majority of the remaining strains were found closely related to Bacillus pumilus, but their average nucleotide identity based on BLAST and electronic DNA/DNA hybridization values excluded closer taxonomic identification. A total of 45 different gene clusters involved in synthesis of secondary metabolites were detected by mining the genomes of the 45 selected strains. Except eight mesophilic strains, the 37 remaining strains were found either cold-adapted or psychrophilic, able to propagate at 10°C and below (Bacillus wiedmannii NMSL88 and Bacillus sp. RJGP41). Pot experiments performed at 10°C with winter wheat seedlings revealed that cold-adapted representatives of B. pumilus, B. safensis and B. atrophaeus promoted growth of the seedlings under cold conditions, suggesting that these bacilli isolated from a cold environment are promising candidates for developing of bioformulations useful for application in sustainable agriculture under environmental conditions unfavourable for the mesophilic bacteria presently in use.     

Ionregulation in juvenile swordspine snook (Centropomus ensiferus,Poey, 1860) in relation to environmental salinity

Ninety‐nine swordspine snook Centropomus ensiferus (9.80 ± 0.3 g, mean ± SE) were studied in order to evaluate the influence of salinity on physiological properties under rearing conditions. Growth performance, survival rates, and ion concentrations (Na⁺, K⁺, Cl^?) as well proximal composition were measured over 76 days. Fish were exposed to three experimental salinities (0, 10, 20 ‰ , three replicates per treatment) and maintained in plastic tanks with a recirculation system equipped with flow‐through aquaria pumps (533 L per tank). Fish were fed twice daily to apparent satiation; at the end of the experiment the weight of fish kept in 10 ‰ was higher than that of fish kept in 0 and 20 ‰ , however no significant differences (P > 0.05) were observed among the experimental salinities. Survival was significantly lower in 10 ‰ salinity than in fish kept in 20 and 0 ‰ salinities. No significant differences (P > 0.05) were found in the Condition factor (K), specific growth rate (SGR), or in plasma Na⁺, K⁺, or Cl^? concentrations among treatments. Salinities also did not affect body composition (P > 0.05), but were significantly lower (P < 0.05) than at the start of the experiment. However, towards the end of the experiment a large accumulation of visceral fat in fish farmed in the three salinities (VFI > 4%) was observed. Water quality was within the optimum range (T: 28.7 ± 0.1°C; O2: 5.6 ± 0.1 mg L^?1; ammonia: 0.2 mg L^?1) for the growth of swordspine snook. Data indicates that C. ensiferus is an ionoregulator fish and able to cultivate successfully in various osmotic conditions, and in turn, maintain high levels of survival in captivity.     

Degradation of Nucleic Acids and their Related Compounds by Microbial Enzymes

Seven strains of microorganisms selected by the previous screening tests were further compared on their ability to produce extracellular enzyme systems capable of degrading RNA into 5′-ribonucleotides. As a result, two strains of Streptomyces were finally concluded to be most preferable. When these two were applied, the rate of 5′-nucleotide production reached up to 70%.

Bacillus subtilis was outstanding in its activity to degrade RNA, but its PDase activity producing 5′-nucleotides from RNA was found to be lower than those of Streptomyces strains. A pathway involving 3′- and 5′-nucleotides as intermediates was proposed for the degradation of RNA by the Bacillus enzyme system. The activity of RNA-degrading enzyme system of Bacillus subtilis contained in the supernatant of culture fluid was found to be lost at 700°C but remained to certain extent at 100°C, a possible mechanism for the phenomenon being discussed. Usability of the Bacillus enzyme system in the practical production of 5′-nucleotides under the condition of high RNA concentration was discussed.     

High-level expression of an endoxylanase gene from Bacillus sp. in Bacillus subtilis DB104 for the production of xylobiose from xylan

To produce xylobiose from xylan, high-level expression of an endoxylanase gene from Bacillus sp. was carried out in Bacillus subtilis DB104. A 1.62-kb SmaI DNA fragment, coding for an endoxylanase of Bacillus sp., was ligated into the Escherichia coli/B. subtilis shuttle vector pJH27Δ88, producing pJHKJ4, which was subsequently transformed into B. subtilis DB104. A maximum endoxylanase activity of 105 U/ml was obtained from the supernatant of B. subtilis DB104 harboring pJHKJ4. The endoxylanase was purified to homogeneity by ion-exchange chromatography and the production profile of xylooligosaccharides from xylan by the endoxylanase was examined by HPLC with a carbohydrate analysis column. Xylobiose was the major product from xylan at 40 °C and its proportion in the xylan hydrolyzates increased with the reaction time; at 12 h, over 60% of the reaction products was xylobiose. These results suggest that xylobiose, which has a stimulatory effect on the selective growth of the intestinal bacterium Bifidobacterium, can be mass-produced effectively by the endoxylanase of Bacillus sp. cloned in B. subtilis. Received: 2 January 1998 / Received revision: 4 March 1998 / Accepted: 4 March 1998     

Restriction and modification in Bacillus species: genetic transformation of bacteria with DNA from different species, part I.

Summary Host specific restriction was detected in 13 Bacillus strains, when 63 strains of Bacillus subtilis and 15 other Bacillus strains were tested with phage 105C. These 13 strains were classified into 8 groups (M,H,C,N,E,F,G,P) by the type of restriction. M-type strains (B. subtilis Marburg 168, its derivatives, and two other strains) showed relatively weak restriction, restricting 105C from other groups of Bacillus by ratios of 10^-1 to 10^-3. Strains of groups H,C,N,E,F,G, and P restricted 105C from other groups by ratios of 10^-2 to 10^-8. It was confirmed with some of the strains that type-specific modification was endowed only by the last host. Furthermore, we isolated one restriction deficient mutant of B. subtilis marburg 168-YS11, which had also lost its modification phenotype.     

水产源芽胞杆菌的分离鉴定及生物学功能分析

筛选具有较好生物学功能的芽胞杆菌(Bacillus),用以改善池塘养殖过程饲料等有机物降解、抑制水体中的病原菌,对从健康养殖鱼虾塘水体中分离的菌株,进行生化试验和16S rDNA序列分子鉴定;通过产酶、耐酸、耐高温试验,抑菌试验以及安全性试验研究分离菌株的生物学功能。试验共筛选出8株细菌,经生化试验及16S rDNA序列的分子鉴定,确定菌株ZHX17、ZHX18、ZHX31为贝莱斯芽胞杆菌（Bacillus velezensis）,菌株ZHX14、ZHX15为地衣芽胞杆菌（Bacillus licheniformis）、菌株NSX4、NSX7、NSX9为枯草芽胞杆菌（Bacillus subtilis）。试验结果表明,8株芽胞杆菌均具有较强的耐酸、耐高温特性,其中3株贝莱斯芽胞杆菌具有较强的分泌淀粉酶、纤维素酶、蛋白酶的能力,抑菌效果好于其他5株芽胞杆菌。安全性试验结果表明,8株芽胞杆菌对草鱼、罗非鱼相对安全。8株芽胞杆菌同时具备分泌淀粉酶、纤维素酶和蛋白酶3种胞外酶的能力,其中贝莱斯芽胞杆菌具有较强的病原菌抑菌能力,可以作为病原拮抗益生菌做进一步研究。     

Induction of systemic resistance in chickpea against Fusarium wilt by Bacillus strains

Plant growth promoting rhizobacteria (PGPR) strains Rb29 (B. amyloliquefaciens MF352007), Bs1 (B. subtilis MF352017) and Bt1 (B. tequilensis MF352019) were tested for growth promotion and for their ability to induce systemic resistance against Fusarium wilt, a vascular disease of chickpea, using two methods that include whole plant and a split-root system. Bacillus strains and Fusarium oxysporum f. sp. ciceris (FOC) were inoculated on separate halves of roots of chickpea seedlings at the same time and then planted in separate pots either in superposition or one side of the other. All Bacillus strains systemically induced resistance against FOC, and significantly (p < 0.05) reduced the wilt disease by 98–100%. Application of Bacillus strains effectively enhanced plant growth, leading to increased plant height, root length, a fresh and dry weight of shoots and roots. These results help to explain the role of strains of Bacillus in growth promotion and biological control of Fusarium wilt in chickpea. This is the first report of systemic-induced resistance against Fusarium wilt in chickpea obtained by application of Bacillus strains to a root system spatially separated from the FOC-inoculated root.