Rhizobium meliloti M14의 니탄배양(泥炭培養)에 관(關)한 연구

Growth and Survival of Rhizobium meliloti M14 on Korean Peat Carrier

전보(前報)에서 분리(分離)한 R. meliloti M14를 공시(供試)하여 국내산 니탄(泥炭)의 이용(利用) 가용성(可用性)을 구명(究明)하고저, 니탄(泥炭)의 중화(中和) 및 개량(改良), 액체배양물과의 혼합(混合) 및 숙성등(熟成等)에 관(關)하여 실험(實驗)하였고, 니탄배양물(泥炭培養物)을 보관(保管)하는 동안 온도(溫度)와 잔존생균수(殘存生菌數)와의 관계(關係)를 검토(檢討)하여 다음과 같은 결과(結果)를 얻었다. 1. 국내산(國內産) 니탄(泥炭)은 산성(酸性), pH 3.8로서 이를 pH6.4로 조성(調聲)하는데 14%의 calcium carbonate가 소요(所要)되었으며, 목탄(木炭), 활성탄등(活性炭等)을 혼용(混用)할 경우에는 4~8%를 절약(節約)할 수 있었다. 2. stationary phase 초기(初期)의 균체배양액을 살균(殺菌)하지 않은 니탄배지(泥炭培地)에 접종(接種)하여 tray배양(培養)할 경우에 3일후(日後) $7{\sim}9{\times}10^9cells/g$의 배양물(培養物)을 얻었으며, 살균(殺菌)한 배지(培地)를 사용(使用)하여 병배양하는 경우에는 5일후(日後) $1.1{\sim}6.2{\times}10^{10}cells/g$의 것을 얻었다. 3. 니탄배양물(泥炭培養物)을 보관(保管)하는 동안, 온도(溫度)에 따라서 잔존생균수(殘存生菌數)의 차이(差異)가 심(甚)하였으며, 특(特)히 $35^{\circ}C$와 $45^{\circ}C$의 높은 온도(溫度)에서 그 감소율(減少率)을 줄이기 위하여 추가적(追加的) 탄소원(炭素源) 및 습윤제(濕潤劑)로서 sorbitol의 첨가효과(添加效果)가 인정(認定)되었다. 본균주(本菌株)는 glycerol, sorbitol, sodium latate 등(等)을 유일(唯一)한 탄소원(炭素源)으로 이용(利用)할 수 있었으나 glycerol 및 sodium latate에서는 생육(生育)이 다소(多少)낮았고, 이들을 0.5%함유(含有)한 니탄배양물(泥炭培養物)을 $25^{\circ}C$에서 보관(保管)할때의 decimal reduction time은 8~9주(週)이었으며, sorbitol첨가물(添加物)의 생균수(生菌數)는 초기(初期)에는 높았으나 12~15주(週) 사이에서 다른 것보다 낮아지는 결과(結果)를 보였다.

This experiment was carried out to study the availability of Korean peat as a main carrier material of rhizobial inoculant, using the alfalfa strain Rhizobium meliloti M 14 which was isolated in the previous report. Modification of powdered peat with calcium carbonate and other materials was studied; inoculation of the peat with culture broth, maturation of the mixture under different conditions, and survival of the strain in the peat culture was examined. The results obtained were as follows. 1. Peat produced in Pyongtak was highly acidic, pH 3.8, and addition of calcium carbonate by 14% was required for pH adjustment to 6.4. However the amount of calcium carbonate could be reduced by 4 to 8% when carbon or charcoal was mixed with the peat. 2. Viable number of the strain reached to $7-9{\times}10^9cells/g$ after 3 days, when inoculated with the culture broth of early stationary growth phase and matured in unsteriled peat of open trays; and the number in steriled peat was $1.1-6.2{\times}10^{10}cells/g$ after 5 days, when matured in closed bottles. 3. Survival of the strain was affected markedly by storage temperature, and positive effect of D-sorbitol on the viability was recognized at elevated temperatures, when added as an additional carbon source and moistening agent. Glycerol, sorbitol, or sodium lactate was utilized by the strain as a sole source of carbon, and the decimal reduction time of viable number in the peat culture was was found to be 8 to 9 weeks at $25^{\circ}C$ when these agents were added by 0.5%.