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＜departmental bulletin paper＞
Quantitative determination of the population of a bacteria by the phage technique

Creator
Creator Name
脇本, 哲
Wakimoto, Satoshi
Affiliation
Affiliation Name
九州大学農学部植物病理学教室
Laboratory of Plant Pathology, Faculty of Agriculture, Kyushu University
Creator Name
吉井, 甫
Yoshii, Hazime
Affiliation
Affiliation Name
九州大学農学部植物病理学教室
Laboratory of Plant Pathology, Faculty of Agriculture, Kyushu University
Language
Japanese
Publisher
九州大學農學部
Faculty of Agriculture, Kyushu University
Date
1955-03
Source Title
Science bulletin of the Faculty of Agriculture, Kyushu University
Vol
15
Issue
2
First Page
161
Last Page
169
Publication Type
Version of Record
Access Rights
open access
JaLC DOI
https://doi.org/10.15017/21356
Related DOI
九州大學農學部學藝雜誌 || 15(2) || p161-169
Science bulletin of the Faculty of Agriculture, Kyushu University || 15(2) || p161-169
http://www.agr.kyushu-u.ac.jp/
Related URI
九州大學農學部學藝雜誌 || 15(2) || p161-169
Science bulletin of the Faculty of Agriculture, Kyushu University || 15(2) || p161-169
http://www.agr.kyushu-u.ac.jp/
Relation
九州大學農學部學藝雜誌 || 15(2) || p161-169
Science bulletin of the Faculty of Agriculture, Kyushu University || 15(2) || p161-169
http://www.agr.kyushu-u.ac.jp/

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Details

Record ID
21356
Peer-Reviewed
Unrefereed
Subject Terms
OP_1 phage
bacteriophage
one-step growth experiment
一段増殖実験
orizae
latent period
ISSN
0368-6264
NCID
AN0005519X
Notes
The multiplication of a specific bacteriophage combined with the host bacterium under a certain environmental condition has a definite passway. Namely, a specific phage-bacterium complex gives always a constant latent period, rise period and average burst size under a certain environmental condition. Based upon these facts, a method was newly deviced to determine the number of　the　living cells of a certain phytopathogenic bacterium within soil and other materials. The method is as follows: 1) The latent period, the rise period and the burst size of the phage-bacterium combination, in question, have at first to be determined. 2) One-Step growth experiment of the phage is then given with the material containing the bacterium in question under multiple infection. 3) Then the number of the living cells of the bacterium (N) will be obtained by the following formula: N= Total number of phage particles produced
Average burst size 4) When the number of the living bacterium is too small to obtain satisfactory results, the second application of the infection of the phage is required, adding the bacterial suspension into the centrifuged supernatant fluid of the phage of stationary period so that the single-infection of the phage is obtained. 5) In the latter case the calculation of the number of the living cells of the bacterium (N’) is as follows: N’=Total number of phage particles produced
(Average burst size)^2 6) The outline of the whole procedure is given in figure 1. Some examples with the application of this method were shown being used Xanthomonas oryzae No.49 and its phage (Op1 phage). From the results, it was found that X. oryzae could survive in the drought soil for a certain period, and that the multiplication within the leaves of rice plants of resistant variety was inferior to that of the susceptible ones.
一般に活性を有するphageと，これにsusceptibleなbacteriaとの接触はphageのbacteriaへの吸着を起し，一定のlatent periodを経て，bacteriaは破壊され，増殖した活性あるphageが放出される．この吸着侵入からlatent periodを経て再びphageを放出する迄の一連の現象は， Delbruckらを嚆矢として，一般に広くphageの増殖現象追求の手段として利用されて来たone-step growth experiment（一段増殖実験)により定量的に究明される．而してbacteriaの生理状態， 培養基の栄養状態及び温度等phageの増殖に債韓影響する環境条件を一定にすると，特定のsusceptible bacteriaと特定のvirulent phageでのone-step growth experimentで明らかにされ得るlatent period， rise period は大体一定であり，更に個々のbacteria から放出されるphage particleの平均数，即ちaverage burst sizeは略一定である．OP_1 Phage(Xanthomonas oryzae bacteriophage)はX．oryzaeの大部分の系統に親和性を有し，而も他の種のbacteriaには作用し得ない．而してOP_1 phageの特定条件下に於ける特定のbacteriaの系統を用いて行ったone-step growth experiment により明らかにされたlatent period， rise period， average burstsizeは既に報告した． 即ち30℃の恒温下でX．oryzae No．49を使用すると， CaVfCh培養基(塩化加里加用無ビタミン･カゼイン加水分解物培養基)中に於けるOP_1 phage のlatent perodは約40分であり， rise periodはそれに続く約20分である．而してaverage burst sizeはmultiple infection(多感染)の場合もsingle infection(単感染)の場合も共に同一で約12を示す．即ちOp_1 phageがX． oryzae No．49 のpopulationに上記の様な条件下で感染した場合，約40分のlatent period を経てbacteriaは破壊され始め，同時にphageを放出し始め，それより約20分の後，個々のbacteriaは完全に，或はpopulationは完全に破壊され，感染したbacteriaの数の約12倍のfree phageを放出し終る．この結果に基きone-step growth experimentの手法を適当に応用すれば，試料中に於て雑菌と混在している状態下に在るX．oryzae No．49をも容易に定量し得る筈である．
Created Date 2012.03.21
Modified Date 2022.11.11

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