• Title/Summary/Keyword: 16SrRNA

Search Result 20, Processing Time 0.172 seconds

Molecular Detection of Phytoplasmas of the 16SrI and 16SrXXXII Groups in Elaeocarpus sylvestris Trees with Decline Disease in Jeju Island, South Korea

  • Geon-Woo, Lee;Sang-Sub, Han
    • The Plant Pathology Journal
    • /
    • v.39 no.1
    • /
    • pp.149-157
    • /
    • 2023
  • Phytoplasmas were discovered in diseased Elaeocarpus sylvestris trees growing on Jeju Island that showed symptoms of yellowing and darkening in the leaves. Leaf samples from 14 symptomatic plants in Jeju-si and Seogwipo-si were collected and phytoplasma 16S rRNA was successfully amplified by nested polymerase chain reaction using universal primers. The sequence analysis detected two phytoplasmas, which showed 99.5% identity to 'Candidatus Phytoplasma asteris' and 'Ca. P. malaysianum' affiliated to 16SrI and 16SrXXXII groups, respectively. Through polymerase chain reaction-restriction fragment length polymorphism (RFLP) analyses using the AfaI (RsaI) restriction enzyme, the presence of two phytoplasmas strains as well as cases of mixed infection of these strains was detected. In a virtual RFLP analysis with 17 restriction enzymes, the 16S rRNA sequence of the 'Ca. P. asteris' strain was found to match the pattern of the 16SrI-B subgroup. In addition, the phytoplasmas in the mixed-infection cases could be distinguished using specific primer sets. In conclusion, this study confirmed mixed infection of two phytoplasmas in one E. sylvestris plant, and also the presence of two phytoplasmas (of the 16SrI and 16SrXXXII groups) in Jeju Island (Republic of Korea).

Phytoplasma specific primer for detection of jujube witches′ broom group(16SrV) in Korea and China

  • Sangsub Han;Lee, Sanghun;Mengjun Liu;Byeongjin Cha
    • Proceedings of the Korean Society of Plant Pathology Conference
    • /
    • 2003.10a
    • /
    • pp.136.2-137
    • /
    • 2003
  • In order to diagnose and differentiate jujube witches' broom (JWB) phytoplasma rapidly, oligonucleotide primer pair, 16Sr(V) F/R, for polymerase chain reactions (PCRs) was designed on the basis of 165 rRNA sequences of JWB phytoplasma. The PCR employing phytoplasma universal primer pair P1/P7 consistently amplified DNA in all tested phytoplasma isolates. But no phytoplasma DNA was detected in healthy jujube seedlings. The nested PCR, the primer pair 16S(V) F/R, about 460 bp fragment, amplified DNA in all tested JWB and related phytoplasmas including LiWB phytoplasma of the 165 rRNA group V, but no DNA amplification was detected from other phytoplasma strains such as group 16SrI (Aster yellows) and group 16SrⅩII (Stolbur group) phytoplasmas in which mulberry dwarf phytoplasma and chrysanthemum witches broom phytoplasma are belonged to, respectively The same results were obtained from both Korean- and Chinese-isolates of JWB. Nested-PCR using phytoplasma universal primer pair P1/P7 and 16S rRNA group V specific primer pair 16S(V) F/R could detect group V phytoplasma rapidly and easily, in particular JWB phytoplasma.

  • PDF

Specific Primer for Detection of Jujube Witches' Broom Phytoplasma Group (16SrV) in Korea

  • Han, Sang-Sub
    • The Plant Pathology Journal
    • /
    • v.21 no.1
    • /
    • pp.55-58
    • /
    • 2005
  • In order to diagnose and differentiate jujube witches' broom (JWB) phytoplasma rapidly, oligonucleotide primer pair, 16Sr(V) F/R, for polymerase chain reactions (PCRs) was designed on the basis of 16S rRNA sequences of JWB phytoplasma. The PCR employing phytoplasma universal primer pair P1/P7 consistently amplified DNA in all tested phytoplasma isolates. But no phytoplasma DNA was detected from healthy jujube seedlings. The nested PCR, the primer pair 16S(V) F/R, about 460 bp fragment, amplified DNA in all tested JWB and related phytoplasmas including ligustrum witches' broom phytoplasma of the 16S rRNA group V, but no DNA amplification was detected from other phytoplasma strains such as groups 16SrI (Aster yellows) and 16SrXII (Stolbur group) in which mulberry dwarf phytoplasma and chrysanthemum witches' broom phytoplasma belong to, respectively. The same results were obtained from both Korean and Chinese isolates of JWB phytoplasma. Nested-PCR using phytoplasma universal primer pair P1/P7 and 16SrV group-specific primer pair 16S(V) F/R could detect group V phytoplasmas rapidly and easily, in particular JWB phytoplasma.

Rapid and Efficient Detection of 16SrI Group Areca Palm Yellow Leaf Phytoplasma in China by Loop-Mediated Isothermal Amplification

  • Yu, Shao-shuai;Che, Hai-yan;Wang, Sheng-jie;Lin, Cai-li;Lin, Ming-xing;Song, Wei-wei;Tang, Qing-hua;Yan, Wei;Qin, Wei-quan
    • The Plant Pathology Journal
    • /
    • v.36 no.5
    • /
    • pp.459-467
    • /
    • 2020
  • Areca palm yellow leaf (AYL) disease caused by the 16SrI group phytoplasma is a serious threat to the development of the Areca palm industry in China. The 16S rRNA gene sequence was utilized to establish a rapid and efficient detection system efficient for the 16SrI-B subgroup AYL phytoplasma in China by loop-mediated isothermal amplification (LAMP). The results showed that two sets of LAMP detection primers, 16SrDNA-2 and 16SrDNA-3, were efficient for 16SrI-B subgroup AYL phytoplasma in China, with positive results appearing under reaction conditions of 64℃ for 40 min. The lowest detection limit for the two LAMP detection assays was the same at 200 ag/μl, namely approximately 53 copies/μl of the target fragments. Phytoplasma was detected in all AYL disease samples from Baoting, Tunchang, and Wanning counties in Hainan province using the two sets of LAMP primers 16SrDNA-2 and 16SrDNA-3, whereas no phytoplasma was detected in the negative control. The LAMP method established in this study with comparatively high sensitivity and stability, provides reliable results that could be visually detected, making it suitable for application and research in rapid diagnosis of AYL disease, detection of seedlings with the pathogen and breeding of disease-resistant Areca palm varieties.

Identification and Comparison of the Nucleotide Sequence of 16S-23S rRNA Gene Intergenic Small SR(Spacer Region) of Lactobacillus rhamnosus ATCC 53103 with Those of L. casei, L. acidophilus and L. helveticus

  • Byun, J.R.;Yoon, Y.H.
    • Asian-Australasian Journal of Animal Sciences
    • /
    • v.16 no.12
    • /
    • pp.1816-1821
    • /
    • 2003
  • Reliable PCR based identification of lactobacilli has been described utilizing the sequence of 16S-23S rRNA intergenic spacer region. Those sequence comparisons showed a high degree of difference in homology among the strains of L. rhamnosus, L. casei, L. acidophilus and L. helveticus whose 16S-23S rRNA intergenic small SR's sizes were 222 bp, 222 bp, 206 bp and 216 bp respectively. The sequence of 16S-23S rRNA intergenic spacer region of L. rhamnosus ATCC 53103 revealed the close relatedness to those of L. casei strains by the homology ranges from 95.4% to 97.2%. 16S-23S rRNA intergenic spacer region nucleotide sequence of L. acidophilus showed some distant relatedness with L. rhamnosus ATCC 53103 with the homology ranges from 40.3% to 41.8% and that with L. helveticus was shown to be 30% of homology, which exists at the most distant phylogenetic relatedness. The identification of species and strain of lactobacilli was possible on the basis of these results. The common sequences among the 17 strains were CTAAGGAA located in the initiating position of the DNA and some discrepancies were found between the same strains based on these results.

A report of 11 unrecorded bacterial species in Korea isolated in 2017

  • Maeng, Soohyun;Kim, Ju-Young;Jang, Jun Hwee;Kang, Myung-Suk;Kim, Myung Kyum
    • Journal of Species Research
    • /
    • v.7 no.2
    • /
    • pp.135-150
    • /
    • 2018
  • Eleven bacterial strains 17SD2_15, 17Sr1_23, 17SD2_13, 17Sr1_31, 17gy_18, 16B15D, 16B02D, 16B04G, 16B01D, 17U4-2 and 17J28-10 assigned to the phylum Proteobacteria were isolated from soil samples collected from Seoul Women's University, in South Korea. The Belnapia species, strain 17SD2_15 was cocci-shaped and pink-colored. The Methylobacterium species, strain 17Sr1_23, 17SD2_13, 17Sr1_31, and 16B15D were short rod-shaped and pink-colored. The Microvirga species, strain 17gy_18, and 16B02D were short rod-shaped and pink-colored. The Oxalicibacterium species, strain 16B04G was short rod-shaped and pink-colored. The Sphingomonas species, strain 16B01D was short rod-shaped and yellow-colored. The Variovorax species, strain 17U4-2 was cocci-shaped and yellow-colored. The Paracoccus species, 17J28-10 was cocci-shaped and orange-colored. Phylogenetic analysis based on 16S rRNA gene sequence showed that strains 17SD2_15, 17Sr1_23, 17SD2_13, 17Sr1_31, 17gy_18, 16B15D, 16B02D, 16B04G, 16B01D, 17U4-2 and 17J28-10 were most closely related to Belnapia soli (with 99.9% similarity), Methylobacterium gregans (99.1%), Methylobacterium isbiliense (99.6%), Methylobacterium oxalidis (99.9%), Microvirga aerilata (98.7%), Methylobacterium aerolatum (99.0%), Microvirga vignae (100.0%), Noviherbaspirillum canariense (100.0%), Sphingomonas desiccabilis (100.0%), Variovorax humicola (99.6%), and Paracoccus acridae (99.1%), respectively. This is the first report of these eleven species in Korea.

SR proteins regulate V6 exon splicing of CD44 pre-mRNA

  • Loh, Tiing Jen;Moon, Heegyum;Jang, Ha Na;Liu, Yongchao;Choi, Namjeong;Shen, Shengfu;Williams, Darren Reece;Jung, Da-Woon;Zheng, Xuexiu;Shen, Haihong
    • BMB Reports
    • /
    • v.49 no.11
    • /
    • pp.612-616
    • /
    • 2016
  • CD44 pre-mRNA includes 20 exons, of which exons 1-5 ($C_1-C_5$) and exons 16-20 ($C_6-C_{10}$) are constant exons, whereas exons 6-15 ($V_1-V_{10}$) are variant exons. $V_6$-exon-containing isoforms have been known to be implicated in tumor cell invasion and metastasis. In the present study, we performed a SR protein screen for CD44 $V_6$ splicing using overexpression and lentivirus-mediated shRNA treatment. Using a CD44 $V_6$ minigene, we demonstrate that increased SRSF3 and SRSF4 expression do not affect $V_6$ splicing, but increased expression of SRSF1, SRSF6 and SRSF9 significantly inhibit $V_6$ splicing. In addition, using a constitutive exon-specific primer set, we could not detect alterations of CD44 splicing after SR protein-targeting shRNA treatment. However, using a $V_6$ specific primer, we identified that reduced SRSF2 expression significantly reduced the $V_6$ isoform, but increased $V_{6-10}$ and $V_{6,8-10}$ isoforms. Our results indicate that SR proteins are important regulatory proteins for CD44 $V_6$ splicing.

A report of 7 unrecorded bacterial species isolated from several Jeju soil samples in 2016

  • Kim, Ju-Young;Jang, Jun Hwee;Maeng, Soohyun;Kang, Myung-Suk;Kim, Myung Kyum
    • Journal of Species Research
    • /
    • v.7 no.2
    • /
    • pp.151-160
    • /
    • 2018
  • Seven bacterial strains, 15J4M-1, 15J13-8, 16MFM10, 15J1-8, SR1-5-4, 15J13-6, and 15J8-11 assigned to the phylum Actinobacteria, Bacteroidetes, and Firmicutes were isolated from soil samples collected from Jeju, Korea. Phylogenetic analysis based on 16S rRNA gene sequence revealed that strains 15J4M-1, 15J13-8, 16MFM10, 15J1-8, SR1-5-4, 15J13-6, and 15J8-11 were most closely related to Bacillus selenatarsenatis $SF-1^T$ (with 99.4% similarity), Brevibacterium luteolum $CF87^T$ (99.5%), Carnobacterium iners CCUG $62000^T$ (99.6%), Exiguobacterium profundum $10C^T$ (99.3%), Larkinella insperata LMG $22510^T$ (99.3%), Pseudokineococcus lusitanus CECT $7306^T$ (99.4%), and Spirosoma endophyticum $EX36^T$ (99.3%), respectively. This is the first report of these seven species in Korea.

In situ isolation and characterization of the biosurfactants of B. Subtilis

  • Akthar, Wasim S.;Aadham, Mohamed Sheik;Nisha, Arif S.
    • Advances in environmental research
    • /
    • v.9 no.3
    • /
    • pp.215-232
    • /
    • 2020
  • Crude oils are essential source of energy. It is majorly found in geographical locations beneath the earth's surface and crude oil is the main factor for the economic developments in the world. Natural crude oil contains unrefined petroleum composed of hydrocarbons of various molecular weights and it contains other organic materials like aromatic compounds, sulphur compounds, and many other organic compounds. These hydrocarbons are rapidly getting degraded by biosurfactant producing microorganisms. The present study deals with the isolation, purification, and characterization of biosurfactant producing microorganism from oil-contaminated soil. The ability of the microorganism producing biosurfactant was investigated by well diffusion method, drop collapse test, emulsification test, oil displacement activity, and blue agar plate method. The isolate obtained from the oil contaminated soil was identified as Bacillus subtilis. The identification was done by microscopic examinations and further characterization was done by Biochemical tests and 16SrRNA gene sequencing. Purification of the biosurfactant was performed by simple liquid-liquid extraction, and characterization of extracted biosurfactants was done using Fourier transform infrared spectroscopy (FTIR). The degradation of crude oil upon treatment with the partially purified biosurfactant was analyzed by FTIR spectroscopy and Gas-chromatography mass spectroscopy (GC-MS).

Detection of Insect Pathogen Serratia marcescens in Protaetia brevitarsis seulensis (Kolbe) from Korea

  • Kwak, Kyu-Won;Han, Myung-Sae;Nam, Sung-Hee;Choi, Ji-Young;Lee, Seok-Hyun;Choi, Young-Cheol;Park, Kwan-Ho
    • International Journal of Industrial Entomology and Biomaterials
    • /
    • v.28 no.2
    • /
    • pp.25-31
    • /
    • 2014
  • Protaetia brevitarsis seulensis (Kolbe) is widely used in Korea, as a protein-rich, alternate, functional food with pharmacological benefits. In addition to anti-oxidant properties, the larvae of P. b. seulensis also show positive effects against hepatic disorder and diabetes; therefore, P. b. seulensis larvae are being reared on a large scale in Korea. We evaluated reared larvae of P. b. seulensis from Gyeong-gi in Korea. Using 16SrRNA PCR, electro-microscopy, and bioassay techniques, we found that the larvae harbored Spo-1, a bacterium identified as the insect pathogen Serratia marcescens. Therefore, we highlight the use of this insect as an alternate food and the need for its sanitary rearing conditions, as contamination may affect public health.