TAP cni CONG THIfONE
PHAN LAP VA KHAO SAT DAC TINH CUA VI KHUAN NOI SINH d CAY NGHE VANG TAI TINH AN GIANG
• LY THI THANH THAO
TOM TAT:
Nghien effu phdn lap vi khud'n ndi smh trong cay nghe vdng trdng d huydn Chau Thanh va TP. Long Xuydn, tinh An Giang nhdm tim dtfdc cac ddng vi khuan ed tinh khang khuan. Trong sd'31 dong vi khuan da phan Idp dffde cho tha'y 16/31 ddngcd boat tinh khang E Co//, 17/31 ddng khang Aeromonas hydrophila, 5/31 ddng khdng Staphylococcus aureus, 'il'i 1 ddng khang ca 3 loai vi khuan gay bdnh. Ngodi ra, ehung cdn cd dae tinh ed djnh dam va tdng hdp lAA. Bdn canh do, 21 ddng vi khuan cd kha nang hoa tan ldn khd tan. Kd't qud BLAST N eho tha'y ddng TT5, RT9, CT7 va CT3 dtfdc xae djnh l^n Itfdt la vi khuan Microbacterium testaceum vdi dd ttfdng ddng 99%, Klebsiella pneumoniae vdi dd ttfdng ddng 99%, Enterobacter cloacea vdi dd ttfdng ddng la 98% va Bacillus pumilus vdi do ttfdng ddng 95%.
Tuf khda: Cay nghd vang, cd' dmh dam, hda tan lan, khdng khuan, tong hdp lAA.
1. Dat van d^
Cay nghd vang cd tdn khoa hoc la Curcuma longa Linn., thudc ho gffng la mdt trong cac loai thtfc vat ed nhieu tae dung. Nghd chffa tinh dau curcumin ra't quy va dffde sff dung lam thud'c rdng khap trdn the' gidi [9], Curcumin vdi nong dO 2,5 - 50 mg/100 mL ngan can dffde sff phdt trien cua Staphylococcus aureus. Cd curcumin vd tinh dau dieu tfc che' sff tang trffdng eua Streptococcus, Staphylococcus. Lactobacillus... [3]. Dung dich nffde trich tff vung re cua nghd vang ed tdc dung khang khuan [II], curcumin ngan ngffa sff phat trien Helicobacter pyloriCag A"^ [14].
Theo Kumar et al. (2015): Tap doan vi khuan ndi sinh phan lap tff cay nghe vdng cd kha nang tdng hdp lAA, hda tan ldn, chiu man, ffc chd' stf phdt trien cua vi khua'n E.Coli, na'm Fusarium solani va Alterneria alternata. 6 Viet Nam, vide
nghidn cffu ve vi khuan ndi sinh d cay nghe vdng cd khd nang kich thich edy phat trien td'i thdng qua kha nang cd dinh dam, tdng hdp lAA, phan gidi lan,^ gdp phan gidm cac loai bdnh, giam chi phi san xud't va dde biet Id khd ndng tdng hdp cac hoat cha't khang khuan cua cde ddng vi khuan khi sd'ng noi sinh vdi eay chffa dtfde quan tdm nhieu. Vi thd', vigc tuyen^chgn, dmh danh va khao sdt dde tinh cua vi khuan ndi sinh d edy nghe vang nhdm phan lap dtfdc nhffng ddng vi khuan ndi sinh ed ich, ffng dung trong san xud'i phdn vi smh, phat trien cay dtfdc lidu cd boat tinh khdng khuan hieu qua Id rd't cap thie't,
2. Phtfdng phap nghien cffu 2.1. Ddi tffang nghien cffu
Toan bd phan than, la, rd vd cu cua edy nghd vdng trdng d J P . Long Xuyen va huyen Chau Thdnh thudc tinh An Giang,
3 1 0 So 7 - T h a n g 4/2019
HOA HOC-CdllG NGHE THUG PHAM
Vi khuan E.Coli vd S.Aureus dtfde eung ca'p tff phong thi nghidm Sinh hgc phan tff, Vien Nghien cihi va Phat tridn Cdng nghd Sinh hgc, Trtfdng Dai hoc Can Thd. Vi khuan A. hydrophila dtfdc cung cap tii' Bd mdn bdnh Cd, Khoa Thuy sdn, Trtfdng Dai hoc Can Thd,
2.2. Phdn lap vi khudn nQi sinh trong cay nghe vang
Rffa sach mau nghd dtfdi vdi ntfde dd loai bo dat, tach rdi re, thdn. Id va cu. Rffa mdu vdi ntfdc cat vd trung vd cat thdnh doan ngan 2 - 3 em. Khff irungbe mat mau bdng cdn 70%, H202 3%, rffa lai bang ntfdc cd't vd trung, Cae mdu sau khi khff Iriing be mat dtfdc edt thdnh mie'ng nho va dtfdc gia iihuyen trong ed'i, cho 1 - 2 mL ntfdc cd't vd ti'iing vao mau, trdn deu.
Hut 0,1 mL djch mau nghien eho vao d'ng nghiem chffa mdi trtfdng NFb bdn dac. Sau khi u d nhiet do SO^C trong 24 - 48 gid, cdc ddng vi khua'n du'dc phdn lap tff re, than va la cua cay nghd phat then tao thanh mdt vdng pellicle trang due ben dtfdi bi mat mdi trtfdng NFb ban dac trong d'ng nghidm khoang 2 - 5 mm, Idp mang nay dac trtfng cho stf phat tridn cua vi khuan vi hie'u khi. Hut 100 [iL dung djch mdu tai vj tri vdng pellicle xua't hien trai deu trdn dTa petri cd mdi trffdng PDA, Ca'y chuyen nhieu iln de tdch rdng vi khuan. Quan sat hinh dang, do kieh thffdc, kha nang chuyen ddng, lien hanh nhudm Gram va ca'y chuyen cdc ddng vi khudn sang d'ng nghiem chffa mdi trffdng PDA dae ttfdng ffng de trff d 4"C.
2.3. Khdo sdt mot sddgc tinh cua cdc dong vi khuan ngi sinh
2.3.1. Khd ndng cd dinh dam
Xac dmh ndng do NH4+ nhd phdn ffng phenol va NH3 vdi sff hidn didn cua tac nhdn oxy hda la hypochloride hinh thdnh cd phffc mau xanh, dtfdi pHkiem, Khao sat kha nang eddinh dam thtfe hidn dtfa trdn phtfdng phdp Indophenol Blue. Dmh ltfdng dam do vi khuan sinh ra ngay 0, 2, 4, 6 (sau khi chung), Dtfa vao sff thay ddi mau giffa thud'c thu' vd NH4+, do phd OD d bffdc sdng 636 nm, ve tren do thj Excel, tinh dtfdc ndng do NH4"^ trong dich VI khuan.
2.3.2. Khd ndng tong hop IPA
Dtfa vao dac diem tryptophan la tien cha't de tong hdp len lAA nen stf hidn didn ciia tryptophan nham xdc dinh hdm Itfdng lAA dtfdc tdng hdp
thdng qua thude thff Salkowski (lAA cho mau hdng nhat vdi FeCl,). Khao sat khd ndng tdng hdp lAA dtfdc thtfe hidn dtfa trdn phffdng phdp Salkowski [5], so mdu d bffdc sdng 530 nm (OD530nm)
2.3.3. Khd ndng hda tan ldn khd tan Cay ehuyen tat ca cac ddng vi khuan ndi sinh tren mdi trtfdng chffa Idn khd tan (NBRIP cdi tie'n) ddc: (g/L): sucrose, 10; Ca3(P04)3, 2,5; MgS04, 7H2O, 0,25; KCl, 0,2; MgC|2.6H20,"5; (NH4)2S04, 0,1; Bromthymol blue 0,5% trong KOH 0,2N, 3;
agar, 18;pH = 7,2) va u d30<'C. [15]
Hieu qud hda tan lan dtfdc tinh theo cdng thtfc:
E% = (dtfdng kinh vdng sdng halo/dtfdng kinh khuan lac) X 100. [17]
2.3.4. Khdo sdt ttnh khdng khudn ciia cdc dong vi khudn noi sinh
Dung gid'y tham dtfdng kinh 6 mm nhung vao dich vi khuan ndi sinh (mat sd' 10^ CPU /mL) trong vai phfft va dat gia'y thd'm len bl mat mdi trtfdng da trai vi khuan gdy bdnh ttfng ddng rieng bidt. Mau dtfdc u d SO^C, 24 gid, Quan sat va ghi nhan ke't qua ve khd nang tfc che' stf phdt trien cua vi khuan gay benh thdng qua hmh thanh vdng vd khuan quanh khud'n lac cua vi khuan ndi sinh [2]
2.4. Nhan dien vd dinh danh vi khudn HQI sinh trien vong
Chgn ddng vi khuan trien vgng thtfc hien phan ffng PCR vdi 2 cap mdi tdng 27F vd 1492R (Lane, 1991): 27F (5'-AGAGTTTGATCCTGGCTC-3');
1492R (5'-TACGGTTACCTTGTTACGACT-3').
Sau khi dd khuech dai DNA cua cac ddng vi khudn, cac ddng vi khuan nay dtfdc gidi trinh ttf va dinh danh, Sff dung edng cu BLASTN tren ngdn hang dff heu gene NCBl de so sanh trinh tff tffdng ddng vdi cac trinh ttf trdn ngdn hdng gen, ddng thdi dtfa vdo cde kd't qua khao sdt ve dac didm hinh thai eua cac dong VI khud'n ngi sinh da phdn lap dtfdc dd nhan dien cdc ddng vi khuan da gidi trinh ttf.
Sd lieu trong cac thi nghiem dffde xff ly bang Microsoft Office Excel 2010 vd phan mem thd'ng kd Statgraphics Centurion XVI.II.
3. Kd't qua va thao lulin
3.1. Ke't qua phdn lap vd dgc diem kliud'n lgc cda cdc dong vi khudn
Ttf rd, than. Id va cii cua mdu nghe vang, 31 ddng vi khuan ngi sinh da dtfdc phdn lap: 3 ddng phan lap tff la, 7 ddng d than, lOdongdcu vd II ddng d rd, Cdc ddng dtfdc ky hidu Id LTx, TTx,
So 7-Thong 4/2019 311
TAP CHi CQNG THIflfNG
CTx, RTx vdi LT, TT, CT va RT lan Itfdt la cdc ddng vi khuan phdn lap dtfdc iff la, than, eu, rd vd X ia sd' thff Iff dong vi khuan.
Cac ddng vi khuan deu phdt tridn ra't nhanh, thdi gian de eac ddng vi khuan phat trien thanh khuan lac la 12 gid, eham nhd't la 24 gid; phan ldn cac vi khuan phdn lap dffde deu ed khuan lae dang trdn, bia nguyen, dd ndi md, mdu trSng due, mdt sd' it khuan lac cd dang khdng deu, bia rang ctfa, bia chia thuy, dp nd'i lai, do ndi phang, mau trdng trong, trang sffa vd mau vang,
Hau hd't cde ddng vi khuan cd dang hinh que, mdt sd' dang eau ddn va dang chudi, 4 ddng Gram dtfdng va 27 ddng Gram dm Trong dd, 22 ddngcd kha ndng chuydn ddng, Chidu ddi vi khuan dao ddng trong khoang 1,29 - 3,43 pm, chieu rdng khoang 0,86-2,14 pm.
Hint! 1: Vi khudn co Gram am (A) va Gram dUdng (B) d vat kinh X100
jML] 0
3.2. Khd ndng cd dinh dgm ciia cdc ddng vi khudn da phdn lap
Nhin chung, eac ddng vi khuan bat dau tao ra Itfdng ammonium d ngay thff 2 nhtfng van cdn tha'p, lu'dng ammonium dtfdc lao ra cao nha't d ngdy thff 4, sau dd bat ddu giam vao ngay thff 6.
Sau khi khao sat kha nang sinh ammonium cua 31 ddng vi khud'n phdn lap, nhan tha'y cd 7 ddng vi khud'n Itfdng ammonium dtfdc tao ra nhieu nhd't. (Bang i).
Khd nang cd djnh dam cua cac ddng vi khuan khao sdt rd't bid'n dong, the hidn stf thich ffng khac nhau cua cdc ddng vi khuan ndy vdi mdi trtfdng khdng dam. Khuynh htfdng nay phu hdp vdi nhtfng md td cua Qin et al. (2014) ve khd nang cd' dinh dam cua vi khua'n ndi sinh d edy Limonium slnense (Girard). Mdt nghidn cffu khac ve cac chung vi khuan ndi sinh Bacillus sp., Pseudomonas putida, B. cereus d edy Nghd vdng cho tha'y cdc ddng vi khuan noi sinh phdn lap dffde deu cd kha nang ed dinh dam [10].
Bdng 1. Lircfng ammonium (pg/mL) do cdc dong vl khuan trien vong tgo ra Dong vi khuan
LT1 TT2 TT5 CT3 CT6 RT4 RT11 CV{%)
Ngay 2 0,53*
o.sy 0,55' 0,55- 0,56' 0,50*
0,48=
4,32 Ngay 4
0,82"
0,80»=
0.87"
0,76' 0,82"
0,88>
0,76"
3,78 Ngay 6
0,40"
0,35=
0,38*=
0,40'»
0,40"
0,36"=
0,41"
5,54 ' Ghi chLi: cac gia In trung binh trong cung mot cottheo sau CO cac mau td giong nhau bleu thi su khac biet khong co y nghia thd'ng ke d mile y nghTa 5% theo phep tbi! Duncan.
3.3. Khd ndng tdng hdp lAA cda cdc dong vi khudn pfidn Igp dffOc
Bdng 2. LUdng lAA (pg/mL) do cdc ddng vi khudn trien vong iqo ra Dong vi khuan
LT1 LT3 TT2 TT5 TT7 CT1 CT6 CT10
RT1 RT4 RT5 RT9 RT10 CV(%)
Ngay 2 23,121 48,77==
39,81' 60,98"
60,67"
50,26=
46,96=
63,48"
49,19'=
51,77' 51,01' 35,93"
63,48"
3,49 Ngay 4
77,77' 79,77"
81,95=
93,44"
79,85=
60,411 66,97' 87,21' 80,46=<i 79,51=' 79,00=' 72,31' 81,08=' 2,59
Ngay 6 7,14*
6.83"
6,96' 6.92' 6.30"
4,47"
5,79' 7,71"
7.24"' 4,74' 7,09' 6.07' 6.98=
5.05
•Ghi chu: cac gia In trung blnb trong ciing mot cot theo sau CO cac mau tu giong nhau bleu thi si/ khac biet khong co y nghia thong keo mdc y nghia 5% theo phep tbil Duncan
312 Sd7-Thang 4/2019
HDA HOC-CONG NGHE THtfC PHAM
Kd't qua khdo sat cho thd'y td't ca cac ddng deu cd kha nang tdng hdp lAA, trong do 13 ddng vi khud'n tdng hdp dffde lAA nhidu nha't. Cdc ddng vi khuan nay ed the tdng hdp dtfdc mdt Itfdng lAA nha't dinh d ngay thtf 2, tang cao nha't vao ngay thff 4 vd giam vao ngay thff 6 sau khi chung vao mdi trtfdng NFb long. Nhieu nghien cffu trdn thd' gidi da phan lap dffde ede ddng vi khuan noi sinh cd kha ndng tdng hdp lAA. Theo kd't qua nghien cffu cua Chen et al. (2014), cac ddng vi khuan ndi sinh trong edy gffng cd kha nang tdng hdp lAA.
Chung Bacillus cereus phan lap tff Sophora alopecuroides ciing cd khd ndng long hdp lAA (0,36 |Jg/ml) [13]. Nghien cffu cua Lata et al.
(2006) trong mdi trffdng ed bd sung Tryptophan, vi khuan Pseudomonas stutzeri dffde phan Idp Iff cay cffc dai tong hdp Itfdng lAA la 18,8 pg/ml.
Chung Bacillus sp. phdn lap tff edy nghd vang cd kha nang tdng hdp lAA [10].
3.4. Kha ndng hoa tan lan kho tan Ke't qua khdo sdt d Bang 3 cho tha'y cd 21/31 ddng vi khu^n cd khd nang hda tan ldn tff dang klid tan thdnh dang dd hd'p thu bieu hien qua dffdng kinh halo. Trong dd, TT5 la ddng cd hieu qud hda tan ldn cao nha't (108,20%), chffng td day la ddng thich nghi vdi mdi trtfdng nhanh, cd kha nang hda tan lan khd tan td't. (Bang 3).
Theo nghien cffu cua Amanda et al, (2017), vi khud'n ndi sinh Aneurinibacillus sp. va Lysinibactllus sp. phan lap tff r i cay ehudi cd kha nang hda tan lan. Cdc chung vl khuan noi sinh Bacillus sp., B.cereus, Pseudomonas putida phdn lap tff cay nghe vang cung cd kha nang hda tan lan khd tan [10],
3.5. Khd ndng khdng khudn ciia cdc dong vi khudn phdn Igp dffde
Thff nghidm kha nang khang 3 ddng vi khuan gay benh cho ke't qua: 16 ddng cd kha ndng khang vi khuan gay benh dtfdng rupt (E.Coli), trong dd ddng CT7 cd dffdng kinh vdng vd khua'n ldn nhd't (9,67 mm); 17 ddng cd kha nang khdng vi khud'n gay benh d cd (A.Hydrophila), CT7 (12,83 mm) Id ddng ed kha nang khang manh nha't; trong 5 ddng cd kha nang khang vi khud'n gay benh trdn ngtfdi (S.Aureus), CT3 la ddng cd kha ndng tao vdng vd khua'n ldn nhd't (4,83 mm).
(Bang 4; Hmh 2),
Bdng 4. Khd ndng khdng khudn cua cdc dong vi khudn noi sinh vdi 3 ddng
vi khudn gay benh Dong
vi khuan LT2 LT3 TT2 TT3 TT5 TT6 TT7 CT1 CT2 CT3 CT5 CT7 CT9 RT1 RT4 RT6 RT7 RT9 RT11
Vong v6 khuan (mm) ECo;/
3,17' 2.008 4,33*
4.33*
7,83' 2,171 6.67'
2,008 6,67=
9,67- 3,83"
3,833"
4,83"
3,33"
7,67' 6.67=
AHydrophila 7,67' 6,17' 5,67' 8.67' 11.67' 2,00' 10.50' 4,335 4,67s 10,33=
12,83"
7.17' 3,00'
10,00=
9.83' 10.33=
12,00'
$.AUIBUS 2,00=
2,00=
4.83' 3,60'
3.50'
'Ghi chii: cac gia tn trung binh trong ciing mot cot theo sau CO cac mau td giong nhau bleu thi su khac biet khong co y nghia thong ke d mile y ngbia 5% theo phep thii Duncan.
Theo nghien cffu eua Kumar et al. (2015), cdc ddng vi khuan noi sinh B.cereus, B. thuringiensis, B. pumilus. Ciavibacler michiganensis phdn lap tff edy nghe vdng cd khd ndng khang lai vi khua'n E.Coli, P.Aeruginosa, na'm Fusarium solani va Alterneria alternata.
3.6. Ke't gild nhdn dien mot so'ddng vi khudn bdng ky thudt PCR
Dffa vao ke't qud khao sat dae tinh, chgn 4 ddng vi khud'n trien vong dd la ddng TT5, CT3, CT7.
RT9 (ed khd nang cd dinh dam, tdng hdp lAA, hda tan lan, khdng vi khu£n cao) thtfc hien phan ffng
So 7 - T h a n g 4/2019 313
TAP CHl GONG IHIfliNG
Bang 3. Hieu qua hoa tan lan (%) cua cdc dong vi khudn npi sinh phan iqp
Dongvi khuan
LT2 TTI TT3 TT5 TT7 CT1 CT3 CT4 CT6 CT7 CT8 CT9 RT1 RT2 RT5 RT6 RT7 RT8 RT9 RT10 RT11 CV("/.)
Th6i gian iJ 1 Ngay
Bkhalo/Bk khuan iac (mm)
0,10 0,26 0,32 0.25 0,36 0,27 0,37 0.11 0.16 0.09 0,17 0,08 0,34 0,30 0,10 0,19 0,24 0,17 0,30 0,17 0,08
Higu qua hoa tan ian
(%)
10,00"
26,75' 32,48=
25,70=
35,18"
27,38' 37,52' 11,11=
15,26«
9,24' 17,16H
8,69' 34,72"
30,04' 10,00' 19,68' 24,72"
17.97»i 30.92' 17.65fli 8.73' 7,04
3 Ngay Dk haio/Dk
khuan iac (mm) 0,26 0,31 0.29 0,50 0,42 0,18 0.44 0.20 0,13 0,34 0,28 0,21 0,29 0.33 0,19 0,23 0,53 0,33 0,39 0,20 0,17
Higu qua hoa tan ian
(%)
26.67*
31,94'"
29,30*
60,00"
42,11"
18,801 44,12' 20,03"
13,85' 34.39' 28,03'«
21.94<
29,26"«
33,33*
19,79' 23,45"
53,63"
33,82' 39,95' 20,86"
17,91' 7,79
5 ngay Bkhaio/Sk
khuan lac (mm)
0,59 0,63 0,55 1,08 0,54 0,52 0.73 0,42 0,23 0,73 0,22 0,24 0,30 0,33 0,11 0,45 0,40 0,18 0,75 0,42 0,48
*Gbi chu: cac gia tn tmng binh trong cdng mot cot theo sau co cac mau tu giong nhau bieu thisdkh y nghia thong ke d mifc y nghia 5% theo phep thii Duncan
tti$u qua hoa tan ian
(%)
59,09"
63,21=
55,71=*
108,20"
54,84=*
52,08'"
73,19' 42,431 23,22' 73,19' 22.82*
24.39' 30,05"
33,43"
11,91' 45,17"
40,888' 18,79"
75,15' 42,87»
48,72"8 9,86 3c biet khong co
314 So 7-Thdng 4/2019
HOA HOC-CONG NGHE THlfC PHAM
Hif}h 2: Kha nang tgo vong vd khudn cua cac dong 112 -TT7 - ns va CT3 khang lgi vi khudn A. hydrophila
PCR vdi 2 cap mdi tdng 27F va 1492R. 4 ddng TT5, CT3, CT7, RT9 dtfdc giai trinh ttf san pham PCR viing gen md hda 16S-rRNA va so sdnh tren ngdn hang dff lidu NCBI cho ket qua nhff sau:
Doan DNA cua ddng TT5 dai 889 bp cd ty le ddng hinh 99% vdi trinh ttf DNA ciia Microbacterium testaceum ddng StL B037. Ke't qua dinh danh phu hdp vdi nghidn cffu eua Tomohiro et al, (2011), boat tinh ffc che AHL eua Microbacterium testaceum ra't cd hieu qua trong viec chd'ng lai eae tae nhdn gdy benh cho eay,
Doan DNA cua ddng CT3 ddi 1462 bp cd ty le ddng hinh 95% vdi trinh Iff DNA ciia Bacillus pumilus ddng SAFR-032. Theo nghien cffu cua Hill et al. (2009), Bacillus pumilus tdn tai trong rudt tdm su (Penaeus monodon), Bacillus pumilus cd kha nang ffc che stf nhidm trung do Vibrio harveyi, V. alginolyticus, V. Parahaemolyticus, ca 3 loai vi khua'n ndy la tac nhdn gay bdnh nguy hai d tdm.
Wei and Wu (1998) da phan lap va nhdn dien dtfde mdt so ddng vi khuan Bacillus licheniformis.
B.Subtilis, B.Cereus, B.Pumilus, ed khd nang cd dinh dam d vtfng rd edy gao.
Doan DNA cua dong CT7 dai 1251 bp cd ty le ddng hinh 98%
vdi tiinh ttf DNA cua Enterobacter cloacae ddng DSM 30054. Enterobacter cloacae cd kha ndng tidt ra cde chd't khang sinh hoac ehd't ddc de chd'ng lai nd'm Pythium ultimum [16], vd na'm Verticillium dahliae. Ben eanh dd, Enterobacter cloacae cung cd khd nang tdng hdp acid phosphata.se hda tan lan trong da't gdp phan thuc day tang trtfdng cho cay trdng [6].
Doan DNA cua ddng RT9 dai 953 bp cd ty le ddng hinh 99% vdi tfinh ttf DNA cua Klebsiella pneumoniae ddng MGH 78578. Theo nghien cffu cua Sara Austin et al. (1990), Klebsiella pneumoniae la ddng vi khud'n cd khd nang cd' djnh dam kha cao. Ngoai ra, Klebsiella pneumoniae dffde phdn lap tff edy lffa mi cung cd kha nang cd'dinh N2 [8]. Ngodi kha nang cd'dinh dam, Klebsiella pneumoniae cdn ed kha ndng tdng hdp kieh thich to tang trffdng thffe vdt (lAA) thue day cay phat tridn [19].
4. Ket lu^n
Tff re, than, la va cii cua cay nghe vdng da phan lap dffde 31 ddng vi khua'n ndi sinh. Ke't qua khao sdt da tim thd'y 16 ddng vi khuan ndi sinh khdng E.Coli, 17 ddng khang A.Hydrophila. 5 ddng khang S. aureus, 16 ddng khdng E.Coli va A.Hydrophila, 3 dong khang cd 3 loai vi khu^ii gay benh. Nhin chung, cdc ddng vi khud'n ndi sinh phan lap deu cd khd ndng cd' dinh dam, tdng hdp lAA, trong do ed 21 ddng vi khua'n cd kha nang hda tan lan khd tan. 4 ddng vi khuan trien vong cd kha ndng khdng khuan manh, cd dmh dam, long hdp lAA va hda tan lan khd tan cao da dtfdc tuyen chgn la TT5. RT9, CT7 vd CT3. Ket qud nhdn dien qua khdo sdt hinh thai vd phdn tieh 16S-rRNA eho thd'y cdc chiing nay ttfdng ddng vdi cde ddng theo thff tff la, Microbacterium testaceum StL B037, Klebsiella pneumoniae M G H 7 8 5 7 8 , Enterobacter cloacea D S M 30054 va Bacillus pumilus S A F R ' 0 3 2 •
S6'7-Thang 4/2019 315
TAP CHi GONG THIfdNG
TAI LIEU THAM KHAO:
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Ngay nh§n bai: 14/3/2019
Ngay phan bi§n danh gia va su'a chi?a: 24/3/2019 Ngay cha'p nhan dang bai: 3/4/2019
Thong tin tdc gid:
ThS. LY THI THANH THAO
Khoa Ndng nghiep - Tai nguyen thiSii nhien Dai hpc An Giang
ISOLATION AND CHARACTERIZATION OF ENDOPHYTIC BACTERIA IN CURCUMA LONGA L. COLLECTED
IN AN GIANG PROVINCE
• MA, LY THI THANH THAO Foculty of Agriculture o n d Natural Resources
An Giang University
ABSTRACT:
This study is to isolate the endophytic bacteria from Curcuma longa L. cultivated in Chau Thanh district, Long Xuyen City in An Giang province in order to find out antibacterial activities.
Thirty-one endophytic bacterial strains vvfere isolated from samples of Curcuma longa L. The study's results showed that 17/31 strains were resistant to Aeromonas hydrophila, 16/31 sti'ains were resistant to Escherichia coli and 5/31 strains were resistant to Staphylococcus aureus.
Among them, there were three strains resistant to all three these pathogens. Almost all sti-ains could fix nitrogen and produced lAA. Besides, 21 strains could solubilize insoluble phosphate.
The TT5 strain yielded highest amount of ammonium (0.87 pg/niL) and lAA (93.44 [jg/mL).
Four strains (TT5, RT9, CT7 and CT3) were chosen for sequensing the 16S-rRNA gene. The BLAST N results showed that the TT5, RT9, CT7 and CT3 strains were belong to Microbacterium testaceum {with 99% similarity), Klebsiella pneumoniae (with 99% simdarity), Enterobacter cloace (with 98% similarity) and Bacillus pumilus (with 95% Mmilant}) respectively.
Keywords: Curcuma longa L, nitf-ogen fixation, phosphorus soluble, antibacterial, synllietic indole acetic acid (lAA),
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