APPLIED AND ENVIRONMENTAL MICROBIOLOGY, Nov. 1982, p. 1246-1247 0099-2240/82/1 11246-02$02.

00/0 Copyright 1982, American Society for Microbiology

Vol. 44, No. 5

SO SNH CC PHNG PHP M MT S RHIZOBIUM SPP. C CHNG VO THAN BN

H. J. HOBEN AND P. SOMASEGARAN* NifTAL Project, Department ofAgronomy and Soil Science, College of Tropical Agriculture and Human Resources, University of Hawaii, Honolulu, Hawaii 96822 Received 9 March 1982/Accepted 15 July 1982 Ngun vi sinh trn vo than bn c m bng phng php , tri, nh git. Kt qu cho thy 3 phng php c th thay th cho nhau. Phng php nh git th c a chung hn v tnh tit kim vt liu v cng lao ng ca n. Trong vic kim sot cht lng phn chng c trn vo than bn, th s lng Rhizobium spp. kh hu c xc nh trn mi trng agar hoc cho nhim vo thc vt. xc nh cht lng phn chng c trn than bn gm cc phng php: a, tri, nh git. Khi sn xut vi qui m ln, vic kim tra rt thng xuyn, nn phng php c chn cn phi tit kim v mi trng v s lng a petri. Bn cnh , phng php nn d thc hin v cho kt qu chnh xc. Phng php v tri a c s dng rng ri m s lng Rhizobium kh hu trong mi trng nui cy v trng. Phng php nh git c s dng t hn v khng c thng tin v hiu qu ca phng php ny so vi m , tri. so snh nhng phng php ny, chng ta kho st trn 10 loi phn chng khc nhau. Mi dng Rhizobium c nui ring trong 100 ml dch mannite trch t nm men trong erlen 250 ml. Erlen c ly tm vi vn tc 100 vng/ pht ri em 280C. Sau em trn vi than bn c mua t Nitragin Co., Milwaukee, Wis., vi pH= 6.5-6.8 v s hin din ca CaCO 3. Tip , 50g than bn c cho vo ti polyethylene (16.5 x 11.5 x 0.0038 cm) v c v trng bng tia (2.5 megarads). Mt dng c nha v trng (18G 1.5 in khong 3.8 cm) ng 40 ml dch nui cy (10 9/ ml)

cho vo ti than bn. Mi ti cn c m thng ri dng keo bt kn. Nhng ti c trn u v 280C trong 1 tun. Trong th nghim, 1g mu c chuyn vo 99 ml dch mannitol c pha long trong l. L c lc 15 pht. Mi dng c chun b 5 pha long so vi ban u. Mi pha long ly 0.03 ml, 0.1 ml, 1 ml ln lt cho phng php nh git, tri, a. Mi trng trong cc phng php m l dch trch nm men v agar cha Congo red (0.25%). Trong phng php nh git, mt a c chia lm 8 phn, mi git (0.03 ml) c chng vo mi phn bng pipette chun. Cc a c kh 280C t nht 2 ngy trc khi nh hoc tri. Agar lm ngui 50 0C ri vo tng a khong 15 - 20 ml. Kt qu vic m s lng vi khun trong phn chng bng 3 phng php c trnh by bng di. Khng c s khc bit ln, khng c phng php no cho kt qu qu cao hay qu thp. y l nguyn nhn c 3 phng php u c s dng ti cc phng th nghim. Bng. So snh kt qu phng php m , tri, nh git

Trong phng php , agar ch ngui 45 0C c th gy cht mt s loi vi khn mn cm vi nhit , v vy c th cho kt qu thp. D vy, khi agar 50 0C th phng php khng cho kt qu thp nht tr mu 4. Trong 3 phng php, nh git c a thch nht v c th m nhiu loi trn cng 1 a.

Hnh. Phng php nh git Nguyn nhn do trong phng php ny mi a c chia lm 8 phn so vi phng php tri hoc th phi chun b 8 a. Hn na, khi tng pha long, 4 long cn 4 a. Phng php ny c th tit kim khng gian mu, cng lao ng so vi 2 phng php cn li. Nhng thun li ny l nguyn nhn m phng php nh git c s dng kim tra cht lng phn chng. Trong trng hp phi m vi khun nhit cao th phng php cho nhim vo thc vt thch hp hn m a - c th c lng chnh xc s lng Rhizobium nhim vo.

TI LIU THAM KHO

1. Clark, D. S. 1967. Comparison of pour and surface plate methods for determination of bacterial counts. Can. J. Microbiol. 13: 1409 - 1414. 2. Miles, A. A., and S. S. Misra. 1938. The estimation of the bactericidal power of blood. J. Hyg. 38: 732 - 749. 3. Vincent, J. M. 1970. A manual for the practical study of root-nodule bacteria. IBP (Int. Biol. Programme) 15: 54 - 58. 4. Wilson, D. O., and K. M. Trang. 1980. Effects of storage temperature and enumeration method on Rhizobium spp. numbers in peat inoculants. Trop. Agric. (Trinidad) 57: 233 - 238.