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PHN LP V TUYN CHN MT S CHNG VI KHUN AZOTOBACTER C HOT TNH NITROGENAZA V SINH TNG HP IAA (INDOL AXETIC AXIT) T T THN BNH K- HA QU- NG HNH SN- TP. NNG
TO ISOLATE AND TO SELECT NITROGENASES ACTIVE BACTERIAL GENERA AZOTOBACTER AND SYNTHESIZE AIA FROM HAMLET BINH KYHOA QUY- NGU HANH SON LAND SVTH: NGUYN KIM ANH, lp 04SM PHM TH NGC ANH, lp 05CSM1 L TH THY HOA lp 05CSM1 NGUYN TH QUNH NH, lp 05CSM1 U TH TNH, lp 04SM GVHD:TS. THU H
Khoa: Sinh- Mi trng, Trng i hc S phm Nng.
TM TT T 30 mu t trng la, trng rau v t b hoang ti thn Bnh K- Ha Qu- Ng Hnh Sn- TP. Nng, phn lp v tuyn chn c 08 chng vi khun Azotobacter c kh nng c nh m, trong c 02 chng c kh nng sinh tng hp IAA. Tuyn chn c 2 chng c hot tnh mnh nht: Chng BK- 6 c nh m mnh nht, chng BK- 5 sinh tng hp IAA mnh nht. C th ng dng 2 chng ny lm phn bn vi sinh kch thch s sinh trng ca thc vt hoc b sung vo cc vng t bc mu, ngho dinh dng nng cao ngun m trong t, ci thin sinh thi t SUMMARY Laterfrom 30 soil sample is different: abandonned land rice cultivar, vegetable-raising,.at Binh Ky hamlet, us have already been isolated and selections to 8 genera have Azotobacter bacteria capable of nitrogen fixation, among them there iss 2 genera be capable to synthesize AIA. can apply those active banian make cropped's growth excitation microbiological fertilizer or additives 2 bacterial genera to ash level of nitrogen exacerbation degraded earth regions

1. M u 1.1. t vn Phn bn vi sinh (PBVS) c nhiu u im ni tri so vi phn bn ha hc, ngoi tc dng nng cao nng xut v cht lng cy trng, tit kim phn v c, gim chi ph sn xut th PBVS cn gp phn quan trng trong vic bo v mi trng v pht trin nn nng nghip bn vng. Tuy nhin tnh hnh sn xut PBVS nc ta vn cha p ng nhu cu thc tin sn xut ca nn nng nghip do quy m sn xut nh, cht lng sn phm cha hon thin v n nh. Do d, nghin cu hon thin v nng cao cht lng PBVS l vic lm ht sc cn thit. Trong , vic tuyn chn, nh gi hot tnh ca cc chng VSV l khu u tin v quan trng trong quy trnh to ra ch phm. Azotobacter l mt loi vi khun (VK) hiu kh, sng t do trong t, chng c kh nng c nh m cao v khng ph thuc vo cy ch. Ngoi c im trn th mt s chng thuc chi ny cn c kh nng sinh tng hp nn IAA (cht kch thch sinh trng thc vt). Chnh nh c im quan trng VK Azotobacter c ng dng rng ri trong cc ch phm PBVS lm tng nng sut cy trng.

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Xut pht t l do trn chng ti chn ti: Phn lp v tuyn chn mt s chng VK Azotobacter c hot tnh Nitrogenaza v sinh tng hp IAA t t thn Bnh K- Ha QuNg Hnh Sn- TP. Nng nhm tm ra cc chng VK Azobacter a hot tnh ng dng cho nn nng nghip ti ia phng. 1.2. Mc ch ca ti Phn lp v tuyn chn mt s chng VK Azotobacter c hot tnh Nitrogenaza v sinh tng hp IAA t t thn Bnh K- Ha Qu- Ng Hnh Sn- TP. Nng. 1.3. Ni dung ca ti - Phn lp, tuyn chn cc chng vi khun Azotobacter c hot tnh nitrogenaza v sinh tng hp IAA (Indol Axetic Axit) t t thn Bnh K- Ho Qu- Ng Hnh Sn- TP Nng. - Nghin cu c im nui cy v hnh thi ca cc chng vi khun tuyn chn. - Nghin cu nh hng ca thi gian v nhit nui cy n s sinh trng, kh nng c nh nit v sinh tng hp IAA ca cc chng vi khun tuyn chn. - Thm d nh hng ca dch nui cy cc chng VK tuyn chn n s ny mm ca ht u en. 1.4 Nhng ng gp mi ca ti ti i su vo nghin cu cc chng VK Azotobacter c kh nng c nh m v sinh tng hp IAA, c th ng dng trong nng nghip lm phn bn vi sinh kch thch sinh trng ca thc vt hoc b sung vo cc vng t bc mu nng cao ngun m trong t, ci thin h sinh thi t. 2. i tng v phng php nghin cu 2.1. i tng - Cc chng VK Azotobacter c phn lp t mt s mu t thn Bnh K- Ha QuNg Hnh Sn- TP. Nng. 2.2. Phng php nghin cu - Phng php thu mu, theo Erogov (1983). - Phng php phn lp v tuyn chn cc chng VK Azotobacter , Erogov,(1983). - Phng php m s lng t bo CFU/ml, theo Nguyn Ln Dng (1982). - Phng php xc nh kh nng c nh m ca VK Azotobacter: - Phng php so mu vi thuc th Nessler, theo L Vn Khoa (1996). - Phng php xc nh hm lng IAA theo Miska v Skovski,1983. - Phng php nghin cu c im nui cy v hnh thi ca cc chng VK tuyn chn, theo Nguyn Ln Dng (1988). - Phng php nghin cu th nghim dch nui cy cc chng VK tuyn chn, Mai Th Hng, (2000). - Kha phn loi ca Bergey (1989), Gauzse v cng s (1983). 3. Kt qu v tho lun 3.1. Phn lp v tuyn chn cc chng VK c nh nit v sinh tng hp IAA 3.1.1. Phn lp v tuyn chn cc chng VK Azotobacter c nh nit T 30 mu t khc nhau (trng la, trng mu, t b hoang) ti thn Bnh K- Ha Qu- Ng Hnh Sn- TP. Nng. Chng ti phn lp c 08 chng VK c kh nng c nh nit, trn MT AT. Qua nghin cu chng ti nhn thy c 08 chng VK phn lp c u thuc chi Azotobacter. Bng 3.1. Hm lng NH4+ trong dch nui cy ca cc chng VK Azotobacter tuyn chn

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STT 1 2 3 4 5 6 7 8

Chng VK BK-1 BK-2 BK-3 BK- 4 BK-5 BK-6 BK-7 BK-8

Hm lng NH4+ (mg/ml) 3,10 2,76 2,98 3,14 1,34 3,28 1,65 1,74

Mc c inh nit Mnh Trung bnh Trung bnh Mnh Yu Mnh Yu Yu

Hnh 3.1: Hnh nh ng ging v phn ng mu ca 08 chng VK tuyn chn vi thuc th Nessler 3.1.2. Tuyn chn cc chng VK c nh nit mnh Sau khi s tuyn c 08 chng VK Azotobacter c nh nit, chng ti tip tc tuyn chn cc chng c kh nng c nh nit mnh da vo phn ng mu vi thuc th Nessler. Kt qu c 3 chng mnh, 2 chng trung bnh v 3 chng yu. Trong chng BK- 6 c kh nng c nh m mnh nht (3,28mg/ml), chng ti chn chng ny lm i tng cho cc nghin cu sau.

Hnh 3.2: Hnh nh ng ging v phn ng mu ca chng BK- 6 vi thuc th Nessler 3.1.3. Tuyn chn cc chng VK Azotobacter sinh tng hp IAA T 8 chng VK phn lp c, chng ti tip tc tuyn chn cc chng c kh nng sinh tng hp IAA, da vo phn ng mu vi thuc th Salkowski. Kt qu, xc nh c 2 chng c kh nng sinh tng hp IAA. Trong chng BK- 5 c kh nng sinh tng hp IAA mnh (4,313 g/ml). Chng ti chn chng ny lm i tng cho cc nghin cu tip theo.

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Hnh 3.3: Hnh dng ng ging v phn ng mu ca chng BK- 5 vi thuc th Salkowski 3.2. c im sinh hc ca cc chng VK tuyn chn 3.2.1. c im nui cy v hnh thi ca cc chng VK tuyn chn Bng 3.3: c im nui cy v hnh thi ca chng VK tuyn chn. c im nui cy v hnh thi Chng VK Ngy xut hin KL Mu sc KL - Non: Trng trong - Gi: Trng c - Non mu trng c, khi gi chuyn mu vng. Hnh dng KL - B mt nhn nhy nht, khi gi th nhn nheo, trn u. - KL li, b mt nhn, nhy nht Nhum Gram - Gram m - Gram m Hnh dng t bo. - Hnh cu

BK- 5

BK- 6

- Hnh que ngn

Hnh 3.4: Hnh nh khun lc v hnh dng t bo ca chng BK- 5 v BK- 6 Trn MT lng, dch nui cy ca c 2 chng u c mu trng c u n, c cn. Khi gi chuyn sang mu nu en, snh cao. 3.3. nh hng ca mt s yu t MT ti s sinh trng v hot tnh ca cc chng VK tuyn chn - Chng BK- 6 c kh nng sinh trng v c nh nit mnh (4,52mg NH4+/ml) trn MT AT sau 5 ngy nui cy nhit t 28-300C. - Chng BK- 5 c kh nng sinh tng hp IAA cao( 6,24g/ml) trn MT nc mmpepton c b sung 0,1% tryptophan sau 5 ngy nui cy lc nhit 300C. 3.4. nh hng ca dch nui cy chng VK BK-5 ti s ny mm ca ht u en Sau 24 h, ht u en c x l bng dch nui cy ca chng BK- 5 nng pha long 10-2 v 10-3. Sau 72h, t l ny mm nng pha long 10-2 vt 23% so vi i chng. Nh vy dch nui cy ca chng BK- 5 kch thch v rt ngn thi gian ny mm ca ht ging. V vy, c th ng dng chng ny x l ht ging trc khi gieo.

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i chng

X l bng dch nng 10-2

Hnh 3.5: Hnh nh ht u en ny mm trong a petri sau 72h 3. Kt lun: + T 30 mu t ti thn Bnh K- Ha Qu- Ng Hnh Sn- TP Nng, chng ti phn lp v tuyn chn c 08 chng VK thuc chi Azotobacter c kh nng c nh m, trong chng BK- 6 c kh nng c nh m mnh nht (4,52mg NH4+). Chng BK- 5 v BK- 6 va c kh nng c nh m, va c kh nng sinh tng hp IAA, trong chng BK5 sinh tng hp IAA mnh hn (4,313g/ml). + T 08 chng VK Azotobacter trn, chng ti chn c 02 chng u dng: - Chng BK- 6 c kh nng sinh trng v c nh nit mnh (4,52mg NH4+/ml) trn MT AT sau 5 ngy nui cy nhit t 28-300C. KL non mu trng c, nhy nht, khi gi chuyn mu vng nu, TB hnh que ngn, bt mu Gram m. - Chng BK- 5 c kh nng sinh tng hp IAA cao( 6,24g/ml) trn MT 5 sau 5 ngy nui cy lc nhit 300C. KL mu trng c, nhy nht, TB hnh cu, bt mu Gram m. TI LIU THAM KHO [1] Nguyn Ln Dng, on Xun Mun, Nguyn Phng Tin, Phm Vn Ty (1972), Mt s phng php nghin cu vi sinh vt hc tp 1. NXB KH-KT H Ni. [2] Nguyn Thnh t, Mai Th Hng (2000), Sinh hc vi sinh vt. NXB GD, H Ni. [3] Egorov N.X. (1983) , Thc hnh VSV ( Nguyn Ln Dng dch). NXB MirMatcva, NXB KH-KT H Ni. [4] Bergeys Manual systermatic Baderiology, vol.4(1989). [5] G.F. Gause T.P Preobrazenskai, M.A.Srensnicora, P.P. Terekhova, T. S. Macsimova Opredelitels actinomycetes, Nauka, M.1983.

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