• Title/Summary/Keyword: Pantoea ananatis

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Expression and Activity of Citrus Phytoene Synthase and $\beta$-Carotene Hydroxylase in Escherichia coli

  • Kim, In-Jung;Ko, Kyong-Cheol;Nam, Tae-Sik;Kim, Yu-Wang;Chung, Won-Il;Kim, Chan-Shick
    • Journal of Microbiology
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    • v.41 no.3
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    • pp.212-218
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    • 2003
  • Citrus phytoene synthase (CitPsy) and ${\beta}$-carotene hydroxylase (CitChx), which are involved in caroteinoid biosynthesis, are distantly related to the corresponding bacterial enzymes from the point of view of amino acid sequence similarity. We investigated these enzyme activities using Pantoea ananatis carotenoid biosynthetic genes and Escherichia coli as a host cell. The genes were cloned into two vector systems controlled by the T7 promoter. SDS-polyacrylamide gel electrophoresis showed that CitPsy and CitChx proteins are normally expressed in E. coli in both soluble and insoluble forms. In vivo complementation using the Pantoea ananatis enzymes and HPLC analysis showed that ${\beta}$-carotene and zeaxanthin were produced in recombinant E. coli, which indicated that the citrus enzymes were functionally expressed in E. coli and assembled into a functional multi-enzyme complex with Pantoea ananatis enzymes. These observed activities well matched the results of other researchers on tomato phytoene synthase and Arabidopsis and pepper ${\beta}$-carotene hydroxylases. Thus, our results suggest that plant carotenoid biosynthetic enzymes can generally complement the bacterial enzymes and could be a means of carotenoid production by molecular breeding and fermentation in bacterial and plant systems.

Growth Promotion of Pepper Plants by Pantoea ananatis B1-9 and its Efficient Endophytic Colonization Capacity in Plant Tissues

  • Kim, Su-Nam;Cho, Won-Kyong;Kim, Won-Il;Jee, Hyeong-Jin;Park, Chang-Seuk
    • The Plant Pathology Journal
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    • v.28 no.3
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    • pp.270-281
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    • 2012
  • The bacteria B1-9 that was isolated from the rhizosphere of the green onion could promote growth of pepper, cucumber, tomato, and melon plants. In particular, pepper yield after B1-9 treatment on the seedling was increased about 3 times higher than that of control plants in a field experiment. Partial 16S rDNA sequences revealed that B1-9 belongs to the genus Pantoea ananatis. Pathogenecity tests showed non-pathogenic on kimchi cabbage, carrot, and onion. The functional characterization study demonstrated B1-9's ability to function in phosphate solubilization, sulfur oxidation, nitrogen fixation, and indole-3-acetic acid production. To trace colonization patterns of B1-9 in pepper plant tissues, we used $DRAQ5^{TM}$ fluorescent dye, which stains the DNAs of bacteria and plant cells. A large number of B1-9 cells were found on the surfaces of roots and stems as well as in guard cells. Furthermore, several colonized B1-9 cells resided in inner cortical plant cells. Treatment of rhizosphere regions with strain B1-9 can result in efficient colonization of plants and promote plant growth from the seedling to mature plant stage. In summary, strain B1-9 can be successfully applied in the pepper plantation because of its high colonization capacity in plant tissues, as well as properties that promote efficient plant growth.

Isolation and Characterization of Phosphate Solubilizing Bacteria Pantoea Species as a Plant Growth Promoting Rhizobacteria (식물 생장 촉진 활성을 가진 인산분해 미생물 Pantoea 종의 분리 및 특성 규명)

  • Yun, Chang Yeon;Cheong, Yong Hwa
    • Journal of Life Science
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    • v.26 no.10
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    • pp.1163-1168
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    • 2016
  • Plant growth-promoting rhizobacteria (PGPR) have gained worldwide importance and acceptance due to their agricultural benefits. These microorganisms are potential tools for sustainable agriculture, with effects on plant growth, biofertilization, induced systemic resistance, and biocontrol of plant pathogens. In this study, four different Pantoea species were isolated from field soil, and their plant growth-promoting characteristics were studied. Based on 16S rDNA gene sequencing analyses, the se were grouped into Pantoea ananatis, Pantoea citrea, Pantoea dispersa, Pantoea vagans and named as Pa1, Pc1, Pd1, Pv1, respectively. All of these strains have their ability for solubilization of insoluble phosphate depending on pH decrease at the range around pH 5 at 1days after inoculation and production of plant hormone indole acetic acid (IAA) with 85.3±16.3 μg/ml of Pa1, 183.9±16.8 μg/ml of Pc1, 28.8±17.3 μg/ml of Pd1 and 114.1±16.5 μg/ml of Pv1, respectively. Pa1, Pc1 and Pd1 also have high activity for production of gibberellin (GA3) hormone with 331.1±19.2 μg/ml of Pa1, 288.5±16.8 μg/ml of Pc1, 309.2±18.2 μg/ml of Pd1, but Pv1 does not. Furthermore, all these species have significantly promoted the growth of the lettuce seedling plants at the range around 32~37% for fresh weight and 10~15% for shoot length enhancement, so that these microbe could be used as a potential bio-fertilizer agents.

Two Bacterial Entophytes Eliciting Both Plant Growth Promotion and Plant Defense on Pepper (Capsicum annuum L.)

  • Kang, Seung-Hoon;Cho, Hyun-Soo;Cheong, Hoon;Ryu Choong-Min;Kim, Ji-Hyun;Park, Seung-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.17 no.1
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    • pp.96-103
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    • 2007
  • Plant growth-promoting rhizobacteria (PGPR) have the potential to be used as microbial inoculants to reduce disease incidence and severity and to increase crop yield. Some of the PGPR have been reported to be able to enter plant tissues and establish endophytic populations. Here, we demonstrated an approach to screen bacterial endophytes that have the capacity to promote the growth of pepper seedlings and protect pepper plants against a bacterial pathogen. Initially, out of 150 bacterial isolates collected from healthy stems of peppers cultivated in the Chungcheong and Gyeongsang provinces of Korea, 23 putative endophytic isolates that were considered to be predominating and representative of each pepper sample were selected. By phenotypic characterization and partial 16S rDNA sequence analysis, the isolates were identified as species of Ochrobacterium, Pantoea, Pseudomonas, Sphingomonas, Janthinobacterium, Ralstonia, Arthrobacter, Clavibacter, Sporosarcina, Acidovorax, and Brevundimonas. Among them, two isolates, PS4 and PS27, were selected because they showed consistent colonizing capacity in pepper stems at the levels of $10^6-10^7CFU/g$ tissue, and were found to be most closely related to Pseudomonas rhodesiae and Pantoea ananatis, respectively, by additional analyses of their entire 16S rDNA sequences. Drenching application of the two strains on the pepper seedlings promoted significant growth of peppers, enhancing their root fresh weight by 73.9% and 41.5%, respectively. The two strains also elicited induced systemic resistance of plants against Xanthomonas axonopodis pv. vesicatoria.

Analysis of Microbial Diversity in Makgeolli Fermentation Using PCR-DGGE (PCR-DGGE를 이용한 막걸리발효에서 미생물 다양성 분석)

  • Kwon, Seung-Jik;Ahn, Tae-Young;Sohn, Jae-Hak
    • Journal of Life Science
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    • v.22 no.2
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    • pp.232-238
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    • 2012
  • Kumjungsansung-Makgeolli$^{(R)}$ is a traditional Korean rice wine that is fermented from traditional nuruk and rice. In this study, we performed the PCR-denaturing gradient gel electrophoresis (DGGE) analysis targeting the 16S and 28S rRNA genes to characterize bacterial and fungal diversity during Makgeolli fermentation. The predominant bacteria in the PCR-DGGE profile during Makgeolli fermentation were Lactobacillus spp. (Lactobacillus curvatus, L. kisonensis, L. plantarum, L. sakei, and L. gasseri), Pediococcus spp. (P. acidilactici, P. parvulus, P. agglomerans, and P. pentosaceus), Pantoea spp. (P. agglomerans and P. ananatis), and Citrobacter freundii; these were identified on the base of analysis of 16S rRNA gene sequences. The dominant bacterium during Makgeolli fermentation was L. curvatus. The predominant fungi in PCR-DGGE profile during Makgeolli fermentation were Pichia kudriavzevii, Saccharomyces cerevisiae, Asidia idahoensis, Kluyveromyces marxianus, Saccharomycopsis fibuligera, and Torulaspora delbrueckii, and these were identified on the basis of analysis of 28S rRNA gene sequences. The dominant fungal species during Makgeolli fermentation changed from P. kudriavzevii at 0-2 days incubation to S. cerevisiae at 3-6 days incubation. This study suggests that PCR-DGGE analysis could be a suitable tool for the understanding of microbial diversity and structure during Makgeolli fermentation.

Denaturing Gradient Gel Electrophoresis and Culture-based Analysis of the Bacterial Community in Cheonggukjang, a Korean Traditional Fermented Soybean Food from Gangwon Province (DGGE 방법과 배양법을 이용한 강원지역 전통 발효 청국장에서 미생물의 다양성 분석)

  • Hong, Sung Wook;Lim, In Kyu;Kim, Yong Woo;Shin, Seung-Mee;Chung, Kun Sub
    • Korean Journal of Food Science and Technology
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    • v.45 no.4
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    • pp.515-520
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    • 2013
  • Bacterial communities derived from cheonggukjang and raw rice straw collected from a Mireuksan farm and a Heungup cheonggukjang in Gangwon province were investigated using both culture-based method and denaturing gradient gel electrophoresis (DGGE) analysis. Pure cultures, which were isolated from raw rice straw and cheonggukjang and cultured on tryptic soy agar plates (53-76 colonies per plate), were identified by analysis of 16S rRNA sequences. The traditional culture-based method and analysis of PCR-amplified 16S rRNA by DGGE revealed that for samples collected from the Mireuksan farm, Pantoea agglomerans and Bacillus subtilis were the predominant species in the raw rice-straw and cheonggukjang, respectively. For samples collected from the Heungup cheonggukjang, Bacillus amyloliquefaciens was the predominant species in both raw rice straw and cheonggukjang. Other microorganisms, including members of Pantoea, Bacillus, Enterococcus, Enterobacter, Pseudomonas, Rhodococcus, and Acinetobacter, were also present in the raw rice-straw and cheonggukjang, as were bacteria that could not be cultured.

Effect on Insecticide Susceptibility of Lissorhoptrus oryzophilus Fed on Carotenoid-Biofortified Rice Variety (비타민 A 강화벼 급이가 벼물바구미(Lissorhoptrus oryzophilus)의 살충제 감수성에 미치는 영향)

  • Oh, Sung-Dug;Lee, Ki-Jong;Park, Soo-Yun;Ryu, Tae-Hun;Kim, Jae-Kwang;Sohn, Soo-In;Kim, Jin-Seo;Ha, Sun-Hwa;Park, Jong-Sug;Ahn, Byung-Ohg;Cho, Hyun-Suk;Suh, Sang-Jae
    • Korean Journal of Environmental Agriculture
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    • v.31 no.3
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    • pp.286-292
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    • 2012
  • BACKGROUND: The carotenoid-biofortified (PAC) rice was generated by inserting phytoene synthase (Psy) and carotene desaturase (Crtl) genes isolated from Capsicum annuum cv. Nockwang and Pantoea ananatis into the genome of a conventional variety of rice (Nakdongbyeo). In our present study, we studied the effects on insecticide susceptibility of Rice Water Weevil (Lissorhoptrus oryzophilus). METHODS AND RESULTS: The L. oryzophilus were fed on carotenoid-biofortified (PAC) rice and its near non-genetically modified (GM) counterparts (Nakdongbyeo) under $25{\pm}1^{\circ}C$, humidity of $60{\pm}5%$, and photoperiod 16L:8D for more than 60 days. Ten adults were soaked in the Clothianidin SC solution for 5 second in different concentrations, and were detected the mortalities after 24, 48 and 72 hours respectively. Every experiment was conducted with three replications. The cumulative mortalities of L. oryzophilus exposed to Clothianidin SC were similar between two types of feed administration. CONCLUSION: The results of this study suggested that carotenoid-biofortified rice might not affect the insecticide susceptibilities of Lissorhoptrus oryzophilus.