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发光酶基因luxAB标记硅酸盐细菌NBT菌株的研究.pdf

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1、?; yluxABS:A % )NBT)*何琳燕* * null 黄为一( 2 j O4,?;+, 20Q ?;3 F 3 F, a= 3 .1oMnullA % )null?; ynullS:cI|null 1001- 9332( 2004) 07- 1241- 04nullms |null Q93nullDS M null AIntroductionof bioluminescence genes into silicate-dissolving bacteria strainNBT. HE Linyan, HUANG W eiyi(Department of Microbiology,

2、Nanjing Agricultural University, Nanjing 210095, China).-Chin. J. Appl.Ecol. , 2004, 15( 7) : 1241 1244.In this study, silicate- dissolving bacteria NBT strain was grown w ith 0. 25% maltose as carbon source, and the r-ifampicin- resistance w as generated by ultraviolet mutagenesis and streak natura

3、lized to 200nullgnullml- 1. Through tr-iparental cross, the luxAB genes w ere introduced into NBT- R200 at the help of pRK2013. The luminescence ac-tivity of the hybrid strain was tested, w hich indicated that all colonies had a high luminescence activity andkanamycin- , tetracyctine-and rifampicin-

4、 resistance. The NBT- R200 cells prepared from initial logarit hmic growt hphage were more likely to be sensitive to foreign DNA, and the maximum translocation frequency w as up to 6. 70null 10- 5. In addition, the optimal mat ing rat io was 1null1null1. The potassium release ability from feldspar a

5、nd the lu-minescence of lux AB genes marked silicate- dissolving bacteria RL85 strain w ere stable, and hence, the RL85 wasavailable to rhizosphere microecology researches.Key words null Silicate- dissolving bacteria, lux marker genes, Triparental cross.* Z g S/ j1 “( j S39805-2|) .* *Y “ .2003- 07-

6、 28 l, 2004- 02- 12 s.1nullnull nullA % ) r + y 3=%), ?s SA hA F F , a dKa Kas ? 11, 17, 18, 20 , “ -Q4.S: ?;“di15 lux,7S:y 8 |4 1 . ?; yluxAB% kV ,S: )RL85c F 3 | 20Q ?_?;3F 3 F, luxAB% ? i,BW_= 3 3 4 ).12437 null null null null null null null null null null null :?; yluxABS:A % )NBT )null null nul

7、l null null null null null null null nullm4null?; yS: )? )KTFig. 4 Potassium release from feldspar by luxAB genes marked silicate-dissolving bacteria and starting strains.nullnull )Non-inoculated with bacteria, nullnull )Inoculated withdead bacteria, nullnull )Inoculated w ith bacteria.4nullnull nul

8、l 4null1nullh G )8 l“a Y, 0null25% T )8! !.4null2nullYVA % )NBT ),MF 3 FB F 200 nullgnullml- 1NBT-R200 ),c?; ylux AB pTR102nulllux AB pRK2013/ ),V7NBT )?; a 5 * a F 3 F. 3 )8! s8%,?C 3 - % sDNA,M qK Vr6null70 null 10- 5,1 1null1null1 aR.4null3null?; S: )RL85 dK ? ? )NBTM, O20Q F 3 F?;+, aA % )= 3 .

9、ID1null Amin H- S, Meikle A,Glover L-A, etal.1993.Plasmid and chromosomallyencoded luminescence marker systems for detection of Pseudomonasfluo-rescence in soil. Mol Ecol,2: 47 542null Beauchamp CJ, Kloepper JW, Lemke PA. 1993. Luminometric analysesof plant root colonization by bioluminesecent pseud

10、omonads. Can J Mi-crobiol, 39: 434 4413null Benizri E, Baudoin E, Guckert A. 2001. Root colonization by inocu-lated plant grow th- promoting rhizobacteria. Biocon Sci Technol, 11( 5) : 557 5744null Cui M-X(a ) , Zhang C-G(f) , Jin S-Y( ) , et al.1996. Use of a xylE marker gene to monitor the surviva

11、l of rhizobiapopulations in soil. Chin J Appl Ecol ( 3 ), 7( 3) : 287 292( in Chinese)5null Cui M-X(a ) , Zhang C-G(f) , Jin S-Y( ) , et al.1997. Effect of matric potential on the survival of luxAB markedRhizobium fredii in soil. Chin J Appl Ecol ( 3 ) , 8( 5) : 519 526( in Chinese)6null Drahos DJ.

12、1991. Field testing of genetically engineered microor-ganisms. Biotechnol Adv, 9: 157 1717 null Green H, Jensen DF. 1995. A tool for monitoring Trichodermaharzianum: the use of a gus transformant for ecological studies inthe rh izosphere. Phytopathology, 85: 1436 14408null Kloepper JW , Beauchamp CJ

13、. 1992. A review of issues related tomeasuring colonization of plant by cacteria. Can J Microbiol, 38:1219 12329null Li Y- G( S) , Zhou J- C( .) . 2003. Root colonization andnodulation of Sinorhizobium fredii HN01DL in Glycine max rh-izosphere. Chin J Appl Ecol ( 3 ) , 14( 8) : 1283 1286( in Chinese

14、)10null Lin J-S(?) , Cui M-X(a ) , Jin S-Y( ) , et al .1994.T ransposon Tn5 used as an identifiable ecological marker ofR.fredii strains: Horizonal transfer frequency of transposon T n5and its influence on the movement of R.fredii T n5 mutant. ChinJAppl Ecol( 3 ) , 5( 3) : 292 298( in Chinese)11null

15、 Lin Q-M( ) , Rao Z-H ( ) , Sun Y-X( ) , et al .2002. Iditification of silicate- dissolving bacterium and its effect ontomato. Sci Agric Sin(S jnullr) , Li J( null .) , Jiang X(null) , et al. 2001.Study on root colonization of w heat by luxAB genes-marked Bacil-lus megaterium AT CC14581. Microbiolog

16、y ( 3Y) , 28( 6) : 1 4( in Chinese)13null Meng S-D( ) , Zhang Z-Z(f6) . 1997. Studies on nodu-lation and efficiency of S. fredii using GUS gene. Chin J ApplEcol ( 3 ) , 8( 6) : 595 598( in Ch inese)14null Molina L, Ramos C, Duque E, etal .2000. Survival of Pseudomonasputida KT2440 in soil and in the

17、 rhizosph ere of plants. Soil BiolBiochem,32: 315 32115null Prosser JI. 1994. Molecular marker systems for detection of genet-ically engineered micro- organisms in the environment. Microbiolo-gy, 140: 5 1716null Samberook J, Fritsch EF, Maniatis T. 1989. Molecular Cloning: ALaboratory Manual. 2nd. e

18、d. New York: Cold Spring Harbor Labo-ratory Press. 19 21, 304 31617null Sheng X-F( 9/b) , Huang W-Y (B) , Cao X-Y(k) .2001. Dissolution of feldspar and potassium uptake by the strainNBT of silicate bacterium. PlantNutr Fert Sci (!g ) , 7( 4) : 459 466(in Chinese)18null S heng X-F( 9/b) , H uang W-Y(

19、B). 2001. Physiologicalcharacteristics of strain NBT of silicate bacterium. Acta Pedol Sin(r ) , 38( 4) : 569 574( in Chinese)19null Sheng X- F( 9/b) . 2003. Colonization of silicate bacterium strainNBT in w heat roots. Chin J Appl Ecol( 3 ) , 14( 11 ) :1914 1916( in Chinese)20null Sheng X- F, He L-

20、Y, Huang W-Y. 2002. The con ditions of releasingpotassium by a silicate bacterial strain NBT. Agric Sci China, 1( 6) : 595 60421null Steenhoudt O, Zhu P, Vande B A, et al. 2001. A spontaneous chlo-rate-resistant mutant of Azospirillum brasilense Sp245 displays de-fects in nitrate reduction and plant

21、 root colonization. Biol FertSoils, 33: 317 32222null Stew art GSAB, Williams P. 1992. lux genes and the applications ofbacterial bioluminescence. J Gen Microbiol, 138: 1289 130023null Wang P(null ) , Fen X-M ( ) , Li F-D( ) . 2001.Colonization of Mesorhizorium huakuii JS5A16L marked w ithluxAB gene

22、s in the rhizosphere of Astragalus sinicus. Acta PedolSin(r ) , 38( 2) : 265 269( in Chinese)24null Wu S-C ( ;=) , Li L-M ( E) , Li Z-G( ) . 1994. Ob-tain ing streptomycin resistance strain from predom inant rhizospherebacteria and its 15N label. J Microbiol( 3Y) , 21( 4) : 195 198( in Chinese)25nul

23、l Zhang B- X(fg) , Zhang P (fnull ) , C hen X-B (k) .2000. Factors affecting colon ization of introduced m icroorganismson plant roots. Chin J Appl Ecol ( 3 ) , 11( 6 ) : 951 953( in Chinese)Tenull , o, 1976 M 3,p V,1V Yr 3s 3 ,?V 2 . E- mail: hlyan0974 yahoo. com. cn1244nullnull 3null nullnullnull null null null null null null null null null null null null null null null null null 15

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