• Title/Summary/Keyword: vortex shaking

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Effect of Physical Pre-treatment of Mature Seed in Callus Formation and Plant Regeneration of Zoysiagrass (잔디종자의 물리적 전처리가 캘러스 형성과 식물체 재분화에 미치는 효과)

  • Ahn, Na-Young;Kim, Yong-Goo;Rahman, Md. Atikur;Bae, Eun Ji;Choi, Su Min;Lee, Kwang Soo;Lee, Byung-Hyun
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.35 no.4
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    • pp.316-320
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    • 2015
  • In order to improve the transformation efficiency of zoysiagrass by increasing the frequency of callus formation from mature seeds and plant regeneration, the effect of pre-treatment with sea sand was examined. Mature zoysiagrass seeds were given 10 min of swelling time before sea sand treatment using a sea sand and seed ratio of 1 : 1 and a vortex shaking speed of 6 (1,000 rpm) for 10 min. The seeds showed increased callus formation that was more than 2 times the rate in the control. In addition, plant regeneration efficiency was also increased when embryogenic callus induced from sea sand-treated mature seeds was cultured in regeneration medium. These results will be very helpful for improving the genetic transformation frequency of zoysiagrass, a recalcitrant monocot grass.

A Simple Method for Sporangial Formation of the Rice Downy Mildew Pathogen, Sclerophthora macrospora

  • Lee, Hyeong-Jin;Han, Seong-Sook;Kweon, Jin-Hyeuk
    • The Plant Pathology Journal
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    • v.18 no.2
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    • pp.77-80
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    • 2002
  • A simple method for sporangial formation of the rice downy mildew pathogen, Sclerophthora macrospora, on infected leaf tissues was developed to facilitate diagnosis of the disease. Freshly infected young leaves showing whitish to yellowish small spots were selected and cut into small pieces about 2-3 cm in length. About 10-20 pieces were surface sterilized in a 100 ml Duran bottle with 40 ml of 70% ethanol by vigorous shaking for 30 seconds. After washing three times with distilled water, the leaf cuts were submerged in 10 ml of Millipore-filtered paddy water and incubated at $20^{\circ}C$ in the dark. After 8-10 h of incubation, the bottle was vigorously agitated on a vortex mixer, Aliquot amount of the suspension, 0.1-1.0 m1, was spread on a slide glass and examined under a light microscope at 50 or 100x magnification. It was found that light and 1% NaClO strongly inhibit sporangial formation of S. macrospora. Meanwhile, the use of freshly infected young loaves and washing with 70% ethanol stimulated sporangial formation of the fungus on rice leaves.

Axenic Isolation and 16S rRNA Gene Sequence of the Cyanobacterium Microcystis aeruginosa in Downstream of Nakdong River (낙동강 하류에 분포하는 남조류 Microcystis aeruginosa의 무균분리 및 16S rRNA 유전자 염기서열분석)

  • 박홍기;정은영;이유정;정종문;홍용기
    • Journal of Life Science
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    • v.12 no.2
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    • pp.158-163
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    • 2002
  • For axenic isolation of the cyanobacterium Microcystis aeruginosa, water bloom at the Mulgum station from the Nakdong River was pretreated by shaking with distilled water. Removal of bacteria was accomplished using antibiotics (150 $\mu$g/$m\ell$ ampicillin and 25 $\mu$g/$m\ell$ neomycin) and colonizing on CB solid medium prepared from 0.7% agarose at 3$0^{\circ}C$ under 40 $\mu$ mol m$^{-2}$ s$^{-1}$ light. Among 26 strains of the Microcystis species, only three strains were axenically established. The three strains were examined by PCR-amplified 16S rRNA gene and 16S rRNA sequencing. The similarities were 99.5 ~100% with M. aeruginosa AF 139292.

The Anti-Sticking Effect of Mixture of Trisodium Phosphate and Citric Acid on Oral Streptococcus species (구강 내 사슬알균 종들에 대한 제3인산나트륨과 구연산의 탈부착 효과)

  • Jung, Choong-Hyun;Cho, Hyung-Hun;Choi, Gwang-Ju;Kang, Seung-Yong;Yang, Nam-Woong
    • Korean Journal of Microbiology
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    • v.44 no.4
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    • pp.289-292
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    • 2008
  • Trisodium phosphate 12 hydrate and citric acid monohydrate mixture showed the strong anti-sticking effect on Streptococcus mutans, Streptococcus mitis, and Streptococcus salivarius, which are adhered to glass beads. Each Streptococcus species was shaking-cultured in brain heart infusion broth containing three glass beads. After 18 hr, glass beads were slightly washed into normal saline by three-pin-pointed pincette. Each three glass-beads set was put into reagent -containing tubes, which have 40 mg of bits of weighing paper for gaining brushing effect as similar as brushing one's teeth. The tubes were shaken by vortex mixer for 10 min except non-oral microbe, Streptococcus agalactiae (5 min). The samples were colony-counted by serial agar dilution method. Experiment was repeated three times for each Streptococcus species. The relative ratios of bacterial de-adherence by reagents were calculated in comparison with normal saline control. The de-adherence degree of citric acid-trisodium phosphate-saline mixture (CTS, pH 6.0) against Streptococcus mutans came to an average of 12.5 times compared with normal saline control. Trisodium-saline (TS, pH 8.4) showed the average of 7.5 times, and citric acid-saline (CS, pH 4.6) showed 6.0 times compared to the control group. The bacterial de-adherence degree against Streptococcus salivarius was each 7.2,2.6 and 2.8 times in above reagent sequence in comparison with saline control. CTS and TS showed 2.4 and 3.4 times of anti-sticking effect on Streptococcus mitis respectively, but CS had no anti-sticking effect on this bacterium. CTS, TS and CS showed 0.7, 0.6, and 0.6 times on non-oral microbe, Streptococcus agalactiae, separately compared with saline control. These results show that oral Streptococcus mutans, Streptococcus salivarius, and Streptococcus mitis, which are causative of dental caries or subacute endocarditis, may be easily removed from oral cavity by CTS mixture. It is conceivable that our experimental results will enable the development of a new conceptive toothpaste to prevent dental caries or subacute endocarditis after drawing teeth.