Aim: To isolate and identify black pepper (Piper nigrum L) associated endophytic bacteria antagonistic to Phytophthora capsici causing foot rot disease. Methods and Results: Endophytic bacteria (74) were isolated, characterized and evaluated against P. capsici. Six genera belong to Pseudomonas spp (20 strains), Serratia (1 strain), Bacillus spp. (22 strains), Arthrobacter spp. (15 strains), Micrococcus spp. (7 strains), Curtobacterium sp. (1 strain) and eight unidentified strains were isolated from internal tissues of root and stem. Three isolates, IISRBP 35, IISRBP 25 and IISRBP 17 were found effective for Phytophthora suppression in multilevel screening assays which recorded over 70% disease suppression in green house trials. A species closest match (99% similarity) of IISRBP 35 was established as Pseudomonas aeruginosa (Pseudomonas EF568931), IISRBP 25 as P. putida (Pseudomonas EF568932), and IISRBP 17 as Bacillus megaterium (B. megaterium EU071712) based on 16S rDNA sequencing. Conclusion: Black pepper associated P. aeruginosa, P. putida and B. megaterium were identified as effective antagonistic endophytes for biological control of Phytophthora foot rot in black pepper. Significance and Impact of the Study: This work provides the first evidence for endophytic bacterial diversity in black pepper stem and roots, with biocontrol potential against P. capsici infection.
The experiment was conducted with three biofloc treatments and one control in triplicate in 500 L capacity indoor tanks. Biofloc tanks, filled with 350 L of water, were fed with sugarcane molasses (BFTS), tapioca flour (BFTT), wheat flour (BFTW) and clean water as control without biofloc and allowed to stand for 30 days. The postlarvae of Litopenaeus vannamei (Boone, 1931) with an Average body weight of 0.15 ± 0.02 g were stocked at the rate of 130 PL m−2 and cultured for a period of 60 days fed with pelleted feed at the rate of 1.5% of biomass. The total suspended solids (TSS) level was maintained at around 500 mg L−1 in BFT tanks. The addition of carbohydrate significantly reduced the total ammonia‐N (TAN), nitrite‐N and nitrate‐N in water and it significantly increased the total heterotrophic bacteria (THB) population in the biofloc treatments. There was a significant difference in the final average body weight (8.49 ± 0.09 g) in the wheat flour treatment (BFTW) than those treatment and control group of the shrimp. Survival of the shrimps was not affected by the treatments and ranged between 82.02% and 90.3%. The proximate and chemical composition of biofloc and proximate composition of the shrimp was significantly different between the biofloc treatments and control. Tintinids, ciliates, copepods, cyanobacteria and nematodes were identified in all the biofloc treatments, nematodes being the most dominant group of organisms in the biofloc. It could be concluded that the use of wheat flour (BFTW) effectively enhanced the biofloc production and contributed towards better water quality which resulted in higher production of shrimp.
In this study, 100 PGPR strains isolated from different varieties of ginger (Zingiber officinale Rosc.) were first characterized for their morphological, biochemical, and nutrient mobilization traits in vitro. The PGPR were also screened in vitro for inhibition of Pythium myriotylum causing soft rot in ginger. Results revealed that only five PGPR showed >70% suppression of P. myriotylum. These 5 PGPR viz., GRB (Ginger rhizobacteria) 25--Burkholderia cepacia, GRB35--Bacillus amyloliquefaciens; GRB58--Serratia marcescens; GRB68--S. marcescens; GRB91--Pseudomonas aeruginosa were used for further growth promotion and biocontrol studies in the green house and field. The green house study revealed that GRB35 (B. amyloliquefaciens) and GRB68 (S. marcescens) registered markedly higher sprouting (96.3%) and lower disease incidence (48.1%) and greater rhizome yield (365.6 g pot(-1) and 384.4 g pot(-1), respectively), while control registered the lowest sprouting (66%), maximum soft rot incidence (100%) and lowest rhizome yield (134.4 g pot(-1)). In the field experiments also, GRB68 (S. marcescens) and GRB35 (B. amyloliquefaciens) registered the greatest sprouting (80% each), markedly lower soft rot incidence (5.2% and 7.3%, respectively) and higher yield (5.0 and 4.3 kg(3)m(-2), respectively) compared to chemicals like Streptomycin sulphate (73.0%, 18.5% and 2.3 kg(3)m(-2), respectively), Metalaxyl-Mancozeb (73.0%, 14.0% and 3.8 kg(3)m(-2), respectively) and control (73.0%, 25.1% and 2.2 kg 3m(-2), respectively). Overall, the results suggested that for growth promotion and management of soft rot disease in ginger, GRB35 B. amyloliquefaciens and GRB68 S. marcescens could be good alternatives to chemical measures. Since, the latter has been reported to be an opportunistic human pathogen, we recommend the use of B. amyloliquefaciens for integration into nutrient and disease management schedules for ginger cultivation.
Zebra Chip disease is a serious threat to potato production. The pathogen, the phloem-limited bacterium 'Candidatus Liberibacter solanacearum,' is vectored by the potato and tomato psyllid Bactericerca cockerelli to potato and tomato. Patterns of pathogen translocation through phloem in potato and tomato plants were examined to determine whether rate or direction of translocation vary by host species or potato cultivars. Two insects were given a 7-day inoculation access period on a single leaf. Weekly, leaves from upper-, middle-, and lower-tier branches were tested for the presence of 'Ca. L. solanacearum' by polymerase chain reaction (PCR). In tomato and potato, 'Ca. L. solanacearum' was detected 2 to 3 weeks after infestation, most frequently in upper- and middle-tier leaves. In potato, the pathogen was detected in leaves on a second, noninfested stem when the stems remained joined via the tuber. Although rates of pathogen movement were similar among potato cultivars, symptoms developed earlier in more susceptible cultivars. Quantitative PCR indicated that bacterial titers were frequently low in tomato and potato samples (<20 genome units per nanogram of DNA). Results establish that, for improved detection, samples should include newly developing leaves and consider that, under low insect pressure, the pathogen may be undetectable by PCR until 3 weeks after infestation.
Endophytic Pseudomonas aeruginosa strain BP35 was originally isolated from black pepper grown in the rain forest in Kerala, India. Strain PaBP35 was shown to provide significant protection to black pepper against infections by Phytophthora capsici and Radopholus similis. For registration and implementation in disease management programmes, several traits of PaBP35 were investigated including its endophytic behaviour, biocontrol activity, phylogeny and toxicity to mammals. The results showed that PaBP35 efficiently colonized black pepper shoots and displayed a typical spatiotemporal pattern in its endophytic movement with concomitant suppression of Phytophthora rot. Confocal laser scanning microscopy revealed high populations of PaBP35::gfp2 inside tomato plantlets, supporting its endophytic behaviour in other plant species. Polyphasic approaches to genotype PaBP35, including BOX-PCR, recN sequence analysis, multilocus sequence typing and comparative genome hybridization analysis, revealed its uniqueness among P. aeruginosa strains representing clinical habitats. However, like other P. aeruginosa strains, PaBP35 exhibited resistance to antibiotics, grew at 25-41°C and produced rhamnolipids and phenazines. PaBP35 displayed strong type II secretion effectors-mediated cytotoxicity on mammalian A549 cells. Coupled with pathogenicity in a murine airway infection model, we conclude that this plant endophytic strain is as virulent as clinical P. aeruginosa strains. Safety issues related to the selection of plant endophytic bacteria for crop protection are discussed.
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