Nghiên cứu chủng xạ khuẩn VN08A12 - Streptomyces toxytricini có tiềm năng ứng dụng trong xử lý bệnh bạc lá lúa và thân thiện với môi trường

Nghiên cứu chủng xạ khuẩn VN08A12 - Streptomyces toxytricini có tiềm năng ứng dụng trong xử lý bệnh bạc lá lúa và thân thiện với môi trường Nguyễn Đình Hải Trường Đại học Công nghệ Luận văn ThS. ngành: Công nghệ Nanô sinh học (Chuyên ngành đào tạo thí điểm) Người hướng dẫn: TS. Nguyễn Đức Quang Năm bảo vệ: 2012

1 u chng x khun VN08A12 - Streptomyces toxytricinng dng trong x nh bn vng Nguyi i hc  Lu ThS.   c m) ng dn: TS. Nguyc Quang o v: 2012 Abstract. Ti    n c   a chng x khu   c s dng trong ving dch bnh b Vii hp  di truyn h. Ti nghing ca chng x khu cng vm (chut bt lp quan tr  y chng x khu t ng sng. Keywords.  sinh hc; Bnh b; Sinh hc;  Nano Content       1.1.  Bnh bvi khun Xanthomonas oryzae pv. oryzae t trong nhng b Vi th gii. Bm o   i 60% t  t nhiu bin    s  bnh : chn l nh,  dit tr vi khunh, [17]. , . X khun  khum  n sn xut t  ch c [20]. Gng c 10000 ch c sn xut t x khuu cht trong s c ng dn xut ru ch ra rng: c 1000 chng x khup mt    ng 10 chng s     ng s sinh tetracycline [2,3]. B ng x khun {nhng chp ng  mt ca Vit Nam [14]} ti Bng chun Vit Nam (Vi 2 sinh hHQGHN) ha hn tit  hc mc khng ch t vi khun X. oryzae pv. oryzae nh b  Vit Nam hin nay. , vic s dng x khu khng ch sinh hc (s dsinh vn phm t  c ch vi sinh vn tm nh bn vu qu n v m ng.         , : u chng x khun VN08A12  Streptomyces toxytricini m ng dng trong x ch bnh bn vng.   n c a chng x khuc s dng trong ving dch bnh b Vit Nam.  Ti nghing ca chng x khung vt m (chut bt p quan tru xanh)  kim tra s a chng x khun vi ng sng.         2.1. T quan chung    2.1.1. P ca x khun trong t     2.2. C       2 2.2.2.        (Chemotaxonomy) 2.2.3.     2.2.5.         16s rRNA  2.3.  2.4. Bnh          2.5.                3.1.1. Chng x khun VN08A12 n ph  3.1.3. Xoo 3.1.4.    xanh        3.1.5. Chum u   nh loi chng x khun 3.2.1.1.   i bm sinh hc 3.2.1.2 .   i b 3.2.1.3.         16S - rRNA 3 3.2.2. Chn lp nht cho kh a VN08A12 3.2.3  3.2.4.  nghim ng ca d khun chut m   4.1VN08A12 4.1.1. m sinh hc Chng x khun VN08A12 ng tt to khun lc ch p t  mt khun ly t tit nh  t qu ng VN08A12   dng thng xon (mi chui xo 2-7   ng. VN08A12 4.1.2.  4.1.2.1. Khả năng đồng hoá và sử dụng các nguồn carbon khác nhau Theo Shirling & Gottlieb (1966), kh    dn carbon c: - S dng mnh ngun carbon (++) khi x khung carbon kim tra bng hoc mi chng (+) cha glucose.  Ảnh SEM ở 15KV x 3.500 thành 5μm  Ảnh SEM ở 30 kv x 3.500 thành 1μm A. Khuẩn lạc VN08A12 B. Tế bào VN08A12 C. 4 -  s dng ngun carbon (+) khi x khu ng carbon kim tra mt so vi chng (-n i chng (+). - nh v kh  dng ngung kim tra x khun ch n mi chng (-) m,    n so vi chng (+). -  dng ngun carbon (-) khi s  n ca x khu   ng kim tra ngang bng ho so vi s i chng (-). Kt qu  b 1.2VN08A12 Ngun carbon Kh  dng ngun carbon D-xylose - D-fructose ++ Cellobiose - L-arabinose + I-inositol  Rhamnose + Sucrose - D-mannitol + Raffinose  Ghi chú: ++: Sử dụng mạnh nguồn cacbon; +: Có thể sử dụng nguồn cacbon ±: Không xác định khả năng sử dụng nguồn cacbon; - : Không sử dụng nguồn cacbon 5 ng cha Cellobiose ng cha arabinose Môi trường chứa xylose Môi trường chứa fructose Đ.c (-) Đ.c (+) chứa glucose 6 ng cha innositol ng cha Rhamnose ng cha Sucrose ng cha Mannitol ng cha Raffinose -)  -D-lactose, -methyl D-glucoside, D-tagatose, lactamide, alaninamide, adenosine, -cyclodextrin, D-arabitol, lactulose,-methyl D-mannoside,D- trehalose, D-lactic acid methyl ester, D--deoxy adenosine, -cyclodextrin, arbutin, maltose, palatinose, turanose, L-lactic acid, L-alanine, inosine, cellobiose, maltotriose, D- psicose, xylitol, D-malic acid, L-alanyl-glycine, thymidine, glycogen, D-fructose, D-mannitol, D-raffinose, L-malic acid, L-asparagine, uridine, inuline, L-fucose, D-mannose, L-rhamnose, 7 acetic acid, methyl pyruvate, L-glutamic acid, adenosine-- monophosphate, mannan, D- galactose, D-melezitose, -hydroxybutyric acid, glycyl-L- glutamic acid, thymidine-- onophosphate, tween 40, D- galacturonic acid, D-melibiose, salicin, - hydroxybutyric acid, propionic acid, L- pyroglutamic acid, uridine-- monophoshate, gentiobiose, -methyl D- galactoside, sedoheptulosan, - hydroxybutyric acid, pyruvic acid, L-serine, fructose-6- phosphate, N-acetyl-D- glucosamine, D- gluconic acod, -methyl D-galactoside, D-sorbitol, p-hydroxyphenyl acetic acid, succinamic acid, putrescine, glucose-1- phosphate, N-acetyl-D- mannosamine, -D-glucose, 3-methyl glucose, stachyose, -keto glutaric acid, succinic acid, 2,3-butanediol, glucose-6- phosphate, amygdalin, m-inositol, -methyl D-glucoside, sucrose, -keto valeric acid, N-acetyl L-glutamic acid, glycerol, D-L--glycerol phosphate, VN08A12   thy rng chng VN08A12  s dng D-fructose, Cellobiose, L- arabinose, Rhamnose, D-n carbon ch y  dng D- t nht (gn bng kh  dng D-glucose). 4.1.2.2. Khả năng đồng hóa Melanin và khả năng chịu muối 8  u mu  Melanin ca VN08A12  nghim v kh       u mui ca chng VN08A12 y ch   chc n mun 4%. T t qu i v   xp loi chng VN08A12 thuStreptomyces sp. 4.1.3. 16S-    08-A12 4.1.3A.       4.1.3B. M. Marker 10kb 1,2. gen 16s -rRNA   16S-    VN08A12        4.1.3  t,  ng t   i l        .            27   1525R,          1.5kb (4.1.3B)                  .              16s-      . ,                           QIAgen. 1.5kb 1kb 9 -            16               1387 Nu, : TGCAAGTCGAACGATGAACCTCCTTCGGGAGGGGATTAGTGGCGAACGGGTGAG TAACACGTGGGCAATCTGCCCTTCACTCTGGGACAAGCCCTGGAAACGGGGTCTA ATACCGGATACGACTGCGGAAGGCATCTTCCGCGGTGGAAAGCTCCGGCGGTGA AGGATGAGCCCGCGGCCTATCAGCTTGTTGGTGGGGTAATGGCCTACCAAGGCGA CGACGGGTAGCCGGCCTGAGAGGGCGACCGGCCACACTGGGACTGAGACACGGC CCAGACTCCTACGGGAGGCAGCAGTGGGGAATATTGCACAATGGGCGAAAGCCT GATGCAGCGACGCCGCGTGAGGGATGACGGCCTTCGGGTTGTAAACCTCTTTCAG CAGGGAAGAAGCGAAAGTGACGGTACCTGCAGAAGAAGCGCCGGCTAACTACGT GCCAGCAGCCGCGGTAATACGTAGGGCGCAAGCGTTGTCCGGAATTATTGGGCGT AAAGAGCTCGTAGGCGGCCAGTCACGTCGGATGTGAAAGCCCGAGGCTTAACCT CGGGTCTGCATTCGATACGGGCTGGCTAGAGTGTGGTAGGGGAGATCGGAATTCC TGGTGTAGCGGTGAAATGCGCAGATATCAGGAGGAACACCGGTGGCGAAGGCGG ATCTCTGGGCCATTACTGACGCTGAGGAGCGAAAGCGTGGGGAGCGAACAGGAT TAGATACCCTGGTAGTCCACGCCGTAAACGTTGGGAACTAGGTGTTGGCGACATT CCACGTCGTCGGTGCCGCAGCTAACGCATTAAGTTCCCCGCCTGGGGAGTACGGC CGCAAGGCTAAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGCGGAGCAT GTGGCTTAATTCGACGCAACGCGAAGAACCTTACCAAGGCTTGACATATACCGGA AAGCATTAGAGATAGTGCCCCCCTTGTGGTCGGTATACAGGTGGTGCATGGCTGT CGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTG TCCTGTGTTGCCAGCATGCCCTTCGGGGTGATGGGGACTCACAGGAGACCGCCGG GGTCAACTCGGAGGAAGGTGGGGACGACGTCAAGTCATCATGCCCCTTATGTCTT GGGCTGCACACGTGCTACAATGGCCGGTACAATGAGCTGCGATACCGTGAGGTG GAGCGAATCTCAAAAAGCCGGTCTCAGTTCGGATTGGGGTCTGCAACTCGACCCC ATGAAGTCGGAGTCGCTAGTAATCGCAGATCAGCATTGCTGCGGTGAATACGTTC CCGGGCCTTGTACACACCGCCCGTCACGTCACGAAAGTCGGTAACACCCGAAGCC GGTGGCCCAACCCTTGTGGAGG  ,             4.1.3. 4.1.3.         16s   10         08A12  16S          Streptomyces toxytricini       100%.   08A12      .             4.1.4                Streptomyces toxytricini. . - hydroxybutyric acid, pyruvic acid, L-serine, fructose- 6- phosphate, N-acetyl-D- glucosamine, D- gluconic acod, -methyl D-galactoside, D-sorbitol, p-hydroxyphenyl. glucose- 1- phosphate, N-acetyl-D- mannosamine, -D-glucose, 3-methyl glucose, stachyose, -keto glutaric acid, succinic acid, 2,3-butanediol, glucose- 6- phosphate,