HOI NGHI KHOA HOC C O N G NGHE SINH HOC T O A N Q U 6 C 2013
O A N H GIA T H A N H PHAN VI SINH VAT TRONG D A T TRONG CHE, CA PHE, H O TIEU d TAY NGUYEN VA TUYSN CHON TO HOP VI SiNH VAT DA CHO'G NANG 06 TAO Che PHAM VI SINH SCl" DUNG1 CHO PHAT f R|6N BfeN VONG CAC LOAI CAY T R O N G T R S N
Nguyen T h l T h u \ N g u y e n Van T h a o \ D o T h j G l m \ T r i n D i n h Man "^i N g u y e n The T r a n g ' , P h a m T h a n h H f i ' , T r i n T h i H o a '
^ Tmng tim phit triin cbng ngh$ cao. Vien Hin lim Khoa hgc vi C6ng nghi Vi$t Nam
^Vi$n Cbng ngh$ sinh hgc, Vi$n Hin ISm Khoa hgc vi Cdng nghi Vigt Nam
T6MTAT
Trong khuSn kh6 d€ lAi uS s6 TN3/C01 chiing tfii da tiSn hinh diSu tra khu hf vi sinh v|t hiSu tdii tfing s5 cfia mf t s6 mau dat trfing j:he, ci phe, h5 tieu fi Tay Nguyen. S6 lugmg vi sinh vjl hiiu fchi tSng s6 trong cic mlu so v6i dSt frSng 6 m&c tnmg blnh (nam moc
^~ 10* CFU/g dit kho, xa khuan ~ lO' CFU/gdat kho v i vi khuin hieu khi - lO' CFU/g dat ldio).,Tuy nhien, m^t dp cfic vi sinh v^t cb ich cho cay trSng thap hon nliiSu so voi cic lo^i dit n6ng nghifp-khSc. Mgl s6 chfing vi sinh v$t c6 dc ho?t_tinh_c6 djnh djm, phin giii lan v4 sinh chit Idch thich sinh truofig thyc vat Axit Indol Acetic da dupc phan l?ip vi tayka chgn tir dit Udng ci phS, chi, h^
tieu. Trong 14 n^u dit duoc kilm tra c6 3 ralu cd su ton tai cua nli6m vi khuan co dinh nito tvr do Azotobacter, 11 mau co sif ton l^i ciia nhdm vi khuin c6 khi niag co dinh niter thu^c Dh6m Azospirillum, 3 miu c6 si; tSn t^i ciia nhom vi khulu c6 d)iib nita thugc siidtnAcetobacter. Visinh v?t phan giii phSt phat kh6 tan c6 m?t troiig c i 14 ralu dit. Cic cliiing my6n chgn da dugc kiem ha ho^it tinh sinh hgc v i so b$ phan loai. Da iuy£n chgn dirge 9 chfing vi khuin c6 kha uang c6 djoh ni to cao. Tmng dd cd 3 chiing thu$c chi Azotobacter, 3 chiing thu^c chi Azospinllum vi 3 chiing thu^c clri Acetobacter. E)a tuyen chgnducc 3 chung vi khuan vi 3 chfing nim m6c c6 kbi nang ptiin giii lan cao. Trong s6 cac chiing dugc tuyen chgn co khi ning c5 dinh dam ho^c phan giii lan c6_S chfing cd kbi ning smh chit kich thich sinh tmdng AIA (tiong d6 c6 3 chirng Azotobacter, I chung i4ceto6ac(erva 1 chfing vi khuan phSn giii En). Tinb ddi khing cua cac th hgp chiJDg dk tao chl phim phSn vi sinh da chfing, da chiic ning su dyng d|ic thd cho tiing lo^ cay tr^ng tr&n da dugc danh gia. 3 chl ptiim vi sinh da chuc nang sfi dyng cho cic ciy cb£, ci phe, hd lieu da dugc t^o ra v i daog tiln hinh danh gii hi|u lyc ciia che p h ^ len cay trong d dieu ki^n in vitro.
Tit khoa: Ci pb&, chi, hh t i ^ sinh chit kich thich sinh trudng AIA, vi khuin cd djnh dam, vi sinh v|t pbin giai lin
M i i D A UTay Nguy§n, vilng cao nguy§n d5l do Bazan mau m& tni phii ciia nubc ta, la vDng co t i l m nang, t h ^ mgnh trong p h ^ t tn^n t$p trung cac cay c6ng nghifp nhir <A phe, che. ho tieu, cao su, d i ^ u . . . Trong nhOng nam tjua da hinh thSnh ra nhOng viing sin xu^t t$p trung quy m d v ^ l a o ra kh6i lu'ong sdn p h i m khd li^n phgc vu cho xuSt khau v a t i i u dung troi^g nu'd'c. Tuy nhiSn, tlnh hinh thyc th cho t h i y , nhieu 'cSy trflng trong diS cci cS phS, ho tieu m | c dCi c6 t i n g tru'dng cao nhirng t h i l u 6n djnh va t i l m an n h i l u rCii ro vo canh lac, djch hgi va c h l l l u a n g nflng san, qui h o g d i tam vT m6 vh c h i i n lu-pc vi thj tnrdng. NguySn nhSn chlnh x u i t phSt lir y l u l6 kem b i n vO-ng trong canh tSc cSy t r i n g da vi dang hi#n h i r u n h i l u nSm qua trong sSn x u i t nong nghiep ciia nubc ta n6i chung va lai Tdy NguySn n6i ri§ng.
Cd phi, ch6, h i fafiu M nh&ng cay trang I3u niSn (chu k^ cgy d phS 25 nSm, chS trSn 40 n a m , h i ti&u vii ch\fc n i m ) vi v^y h f sinh thdi trono d i t ii tirong doi on dinh. Trong h# sinh thai dat vai tro cilia vi sinh v | t (VSV) la r i t quan trgng, (ISn quan d i n v i | c trao flol cac c h l l dinh duiSng va d u tnic dat.
Tnang khufln k h i a l i ^ i ma so TN3/C01 chdng tfli da t i l n hSnh dieu tra khu h# VSV h i i u khi t i n g s l a i a m $ l s l m l u dat t r i n g ch6, ca phe, h i tifiu; Tien hanh phdn l | p va l u y i n chpn cac chung vl sinh cb cac hogt tlnh c l djnh d^m, phdn giai ian vi sinh c h l l kich thlch sinh Inrong thirc v^t Axit Indol Acetic (AIA) tCr cdc mau dat trong ndy; Tgo c h l p h i m phan vi sinh da d i i i n g , da chii'c nang sCr dgng d | c thii cho cic cSy che, ca phe, ho tieu.
N G U Y £ N L I ^ U V A PHUC7NG PHAP Nguyen l i f u
4 m l u d i t t r i n g ch6 ( C l , C2, C l 1, C12), 4 m l u d i l t r i n g cd phfi ( C F 1 , CF2, CF7, CF8) va 6 m i u d i t t r i n g K l ti&u ( T C - N ^ , T C N . J C C . G 3 . HT4, HT14) d u ^ c thu thap 10 9/2011 - 9/2012 tgi Tay Nguyen. M l u sau khi thu duo'c bao qudn & 4
"C cho den khi tiln hanh phSn Uch.
P h u v n g ' p h a p
Do pH ait, xac dinh 3^ am cba dit, Nguyen LSn Dung vi d i n g t^c gid (1982).
Xic djnh m$t db vi sinh v$t hieu khi tong so trong dit, N g u y i n Lan Dung vd d i n g tdc gid (1982).
- Xdc ^ n h s l lirpng vi k h u i n h i i u khi long s l trfin moi tru&ng thgch dfa MPA - Xdc dinh s l l u p n g xa k h u i n t i n g so trfin moi tnrcrng t h g d i dia Gauze - Xdc dinh s l l u p n g o l m moc t i n g s l trfin mol tarfl'ng Zapekdox.
Phan l^p, tuyen chgn vidinh gii ho^t b'nh vl khuan co djnh dam
• Phan l$p vi W i u i n thuOc nhom Azotobacter trfin mfli tnro'ng Ashby Mannitol Agar theo Tejera vd d i n g tdc gia (2005) - Phdn U p vi khuan thu$c nhom Azospirillum tren mfli truo'ng thgch ban long malate vd mfli tnr&ng thgch Rojo Cflng gfl theo Tejera vd fling tdc gia (2005)
HOl"M&H| KHOA HOC CQNtJ NGHg SINH HQC T O A N Q U O C 2UT3
- Phan Igp vi k h u i n thuflc nh6m_A(;efofcactef tren mfli h w n g LGI_ theo Jimenez va dAng_ta_c_gia (1997)
^TCairajnrticf9t-tmh'"C"o^"f1tfN"cuTl?iithutn"dua ti-fin viec ddnh gia kha nang tilTNHJ* vao mCTTnj&iig nufli cay bang phuong phdp so mau theo Tumer vd Gibson (1980).
P f i l n l$p, tuyin chgn va danh gli hoa,t tfnh v i sinh vat phan giii lan
' Phdn Igp V S V phdn giai lan trgn mfli bxri-ng Genesen theo Babenko va dong tdc gia (1984)
- Xac flinh hogt tinh phan gidi Idn tren mfli t a r i n g thgch theo Babenko v d fling tdc gia (1984) vd txing rtioi tiip&hg d|ch t h l theo Watanabe vd Olsen (1965).
Xic djnh khi nang sinh chit died hba sinh tnrbng thuv v^t axft indol axetic (AIA) tho ciia chiing tuyin chgn theo Brie vd fling tdc gid (1991).
T^a chi phim vl sinh da chbv ning: C h l p h i m flupc sdn x u i t theo nguyen lac nufli d y trfin moi tiuo'ng x I p thanh trOng.
K^T QUA V A T H A O LUiiiN
14 m l u flat t r i n g chfi, ca phfi, ho tifiu cCia Tdy Nguygn da d u p c thu thap. Cac m l u flat d l u l h u | c loai flit do bazan phfi hpp cho t r i n g cdc logi cay cflng nghi#p dai ngdy. pH vd 3$ I m ciia cac m l u flat d u p c tien hanh phan t l c h , Xic djnh pH vi db im cac mau dit
Bang 1. Q$ pH vd d$ i m cCia c i c m i u d i l du'p'c k i l m tra M i u
p H d i t S$ ^m (%)
D i t trfing cS phfi j B i t trong chfi CFI
5,2 ,22,8
CF2 5,0 22,9
CF7 5,5 22,4
CFa 5,2 24,5
C1 5.2 22,6
C2 5,1 18,4
C I I 5.4 22.0
C12 5.7 27,5
DiltrSngh&tieu TCw:
5.5 27,4
TCN 5,6 22,4
TCO 4,8 24,2
G3 6,0 19,2
HT14 5,8 23.5
HT4 5.3 21,8 K i t qud trong B i i i g 1 cho I h l y cdc mau d i t trong ca phfi, ho tieu. che b Tdy Nguy§n d l u thu^c loai chua nhg, khflng thlch hpp vdi sinh II cOa cay trong vi v | y se Idm anh hirdng 161 nang suat cOa cdy. So vol dat t r i n g noi chung dO l m cCia cdc m i u d i l d u p c phdn tich khong cao.
Q l l u tra khu h? VSV hieu k h i t o n g s o
CDng vdi c h i t hij'u ccr, VSV s i n g trong flit cfl •j nghTa quan trpng v f l i cdy trong. Hau n h u moi qud trinh xdy ra trong d i t d l u c l sg ^ a m gia t n / c tilp hogc gidn t i l p ciia VSV (miin hoa, khodng hfla c h i t hOu ccr, phdn gidi, gidi phflng chat dinh dudng vfi c c ti> hpp c h i t khfi tan ho^c t i n g hpp c h i t dinh dufl'ng 10 mfli tu'O'ng...). Chiing tfli fli lieu hdnh d i l u tra khu hg VSV hieu khi long s l va m$t s l nhflm VSV hiru Ich tidng cac m i u dat.
K l l qud flilu tra khu h$ VSV t i n g s l trong cdc m l u d i t d u p c trinh bdy d Bdng 2.
Bang 2. S6 iugng vt sinh vat tong so Irong cac m l u dat (CFU/g trong lirg'ng khd)
MSu
VlJthuIn HKTS (xlO°) . NlmmAcTS
(xlO*) .
• Xg khuin TS ' (X10=)
Dal b i n g ca phd .CFI
4,9
3,5 5,7
CF2 4,1 1,8 2.6
CF7 5.5
3.6 4.8
CF8 6,5
4,2 3.3
D i l trong che C l j C2 5.7
3.4 0.8
4,8
2,7 0,71
C11 6,7
2,5 1,6
012 7,2 3.4
2.5
D i t Irons h i Heu T C H ^
0,91 2
2.74 TCN
1,3 2,2
3.25 TCc j G3
1,1 1,43
2,1 1,9
2,18 2,59 HT14
5,2 3,2
1,2 HT4
1,2
2,8
2,9
Ket qud phdn tlch mgt flO vi k h u i n h i i u khi t i n g so (VKHKTS) ti'ong cac mSu dat b Bang 2 cho I h l y flit trong chfi.cd s l lirpng VKHKTS cao n h l l , sau do flen d i t t r i n g ca phfi va flit trong h i lifiu la nghfio V K H i a S n h i t So l u p n g VKHKTS d cdc d i t ndy Id thugc logi trung blnh so vdi nhieu viing dat fld d u p c chiing tfli phdn Kch trudc fldy. 6" t i t cd cac m i u m%t do n i m moc khoang 10* CFU/g flat khfl. S6 l u p n g n i m moc tong s l tnjng cdc mSu flit d u p c phgn tich t u o n g d u a n g v d i s i l u p n g n i m m i c h-ong a l l nfli chung. K i t qud phan tich m a t f l g x a k h u i n long s l t i ^ n g cdc m l u " d i l d Bang'2"cho t h i y m l u flit t r i n g cd phe cfl s l lupng xa khuan cao Jio'n ti-ong a l l trong ho lifiu, flat t r i n g chfi cfl s | l u p n g x g k h u i n l i n g s l U i l p n h l l . Mgt dp xg k h u i n l i n g s i trong cdc mau d i t b^ng d Tay Nguyen thugc mdc flfl'trung blnh.
Phan l^p vd t u y i n c h p n v l k h u a n c6 d j n h flam
Niter id nguyen l l dinh d u d n g quan trgng v d i cay t r i n g . Hang nam cdy ti^ng i l y fli t d flit hdng trdm t r i f u t i n nito. Bang cdch b i n phan con n g u d i tra Igi cho a l t d u p c khoang > 4 0 % , l u p n g tiiieu hyt cdn Igi c o ban d u p c b l sung bdng niter do hogl flOng s i n g cua VSV (Elshanshoury, 1995). VI vgy vific nghien c u u , s d dyng nguon flgm sinh hgc ndy flupc xem Id m$t gidi phdp quan hpng trong nSng nghigp.
B&ng 3. SA lu>png mgt s l nhdm vl k h u i n c6 djnh d^m trong c i c m l u dat (x10'CFU/g trgng twgng khd]
NhdmVK C l d j n h dgm Azototiacter Azospinllum Acetobacter
D i t trong c i phg ' C F I
0 3,27 4.25
.CF2 0 1,82 5,45
CF7 36,7 3,25 3,33
CF8 0 3,4 0
D i t trong che Cl
0 0 1,85
C2 0 0 3.75
C11 32,5 1,2 3.2
C12 0 0 2.25
Dat Irong h i lifiu TCr,.c I TCN
0 0 23,6 2,25
0 1 0 TCo
0 28,2
0 G3
0 1,75
0 HT14
1.25 25 0
HT4 0 2,25 4,32
HOI NGHi KHOA HQC C O N G NGHE SINH HQC T O A N Q U 6 C 2013
C I I HT14 CF7 Hlnh 1. Mgt so mSu d i t c6 s y t i n tgi cua vl k h u i n c l djnh nlto' t v
do Azotobacmr V i k h u a n c o d j n h N i t a Azotobacter
Vl k h u i n (VK) flupc dng dyng n h i l u trong nflng nghiep Id Azotobacfer. Trong 14 mau a l t d u p c k i l m tra (Bdng 3) chi c f l 3 m l u (Hinh 1) xdc flinh tiily cfl VK Azotobacter, cdc m i u c6n Igi khflng t h i y s u t i n tgi cOa nhflm V K nay, ngay ca khi phdn l i d i tnj-c tifip tir a l t khong pha loang. S d dT n h u vay la do Azoto6ac(erch? thlch ung d pH 7,2 - 8,2, nhigt d$ 28 - 30° C, dO I m 40 - 60%, trong khi cac m l u d i t d u p c
" • • " " " ' •' ' : v$y V K . phdn tich fliu co pH v d d g i m tiilp Azotobacter Idiong ton tgi.
T u 3 m l u chiing tfli da phdn lap d u p c 6 chiing VK Azotobacter. T f i bdo VK nay co hlnh cdu, gram am, kich tiiudc l l bdo dao dpng t d 1.5 - 5.5 p m , k h u i n Igc dgng S mdu t f l n g trong, l l i , nhdy. Khi gid khuan Igc cfl mau vdng lyc hogc mau ndu t h i m . Cac d u i n g ndy d u p c chiing tfli phdn tfch hogt linh d djnh d g m d y a tren khd nang tiet NH4* vdo mfli trudng. Ket qua cho I h l y chOng c l hoat Unh tilt N K i * ttr 135 - 192 \iMlmg trong l u p n g khfl tfi bdo sau 48 h nufli c l y . 3 chClng cfl
hoat tlnh cao thich dng v d i sinh l i cOa t d n g log! cdy d u p c chOng t i i tuyen chon. D y a ti'fin cdc ago d i l m hinh thdi, sinh II, sinh hod 2 chiing Ab-C 11.1 v d /yj-CF 7.2 flupc xdc djnh thugc loai A. chrococcum va 1 chung A b - H T 14.2 tiiugc lodi A. vinelandii. ^ >
VI k h u i n CO d j n h N i t c Azospirillum
Theo Jimmenez vd d i n g tdc gid (1997) l l n s u i t Azospirillum thu flupc trong hg r l khodng 20 - 4 0 % . Azospirillum thlch hpp vdi a l t chua nhg, VK Azospirillum smh tiu'dng tren mfli I n r d n g thgch bdn long malate, tao dfa sinh t r u d n g t i l n sdt b i mgt sau 4 8 - 96 h. T l bdo hlnh cong nhe,
vgn flgng x o l n , c h d a hgt PHB, kich Uiifdc 1 x 3 pm. Trfin mfli t n r d n g thgch flgc higu Rojo Cflng gfl, k h u i n Igc Azospirillum r n d u dd Ua sau 4 ngdy nufli c l y d 37° C. " ~ "
Ti> cdc mdu flat d u p c (dim ta-a (Bang 3) 25 chiing du'pc tdch v d d u p c phdn tlch khd ndng t i l l NH4* vdo mfli t n r d n g sinh t r u d n g b i n g phuang phap so mau. Cdc chfing chpn loc c6 khd ndng l i l t NH4* vdi nhirng lirpng khdc nhau t d 72,55 |JM - 225,75 pM/mg tipng lirpng khfl t l bdo. 3 chiing Azospirillum sp. (Hinh 2) cfl hogt tlnh cao thich dng v d i sinh II cOa tCmg Iogi cdy flupc chiing'lfli t u y i n chgn.
VI khuan c l d j n h Nito- Acetobacter K l l qud phan tich (Bdng 3) cho t h i y tiong 14 m l u flit flupc k i l m tia chi 8 m l u t h i y cd s y ton tg cua VK Acetobacter. S l l u p n g nhflm VK Acetobacte tim tiily ti'ong cac m i u flit nfli chung t h i p mat 6Q chl khodng 10^ CFU/g flit khfl." Theo J m a n e z v d fling tdc gia (1997) Acetobacter d u p c tdch t u d i t t r i n g cd phfi vdi t i n s u i t gap t d 15 den 4 0 ^ tuy thugc vdo pH a l t .
As-C 1.3 As-CF 1.5 As-HT 14.1 Hlnh 2. Cdc chiing Azospirillum 6irgc t u y i n ch^n tir c i c m l u dSt
t r i n g Tay Nguyfin
Ac-C 2 5 Ac-CF 2.2 Ac-HT 4.1 Hlnh 3. C i c chung Acetobacter dugc tuyIn chon t d c i c m i u d i t
trdng Tay Nguy§n T d 8 m i u d i t cliiing 161 da phan Igp flupc 16 chiing
VK Acetobacter. K l l qud cho tiily chiing co kha nang t i l l NH4' t d 27.5 - 1 2 2 , 3 pM/mg trpng l u p n g kho t l bdo. 3 chOng (Hinh 3) cfl hogl tlnh cao thich dng v d i sinh li ciia tdng leal cdy d u p c chiing toi t u y i n chgn. D y a tren cac dgc d i l m h l n h thdi, sinh II, sinh hod xdc flinh flupc cd 3 chung fliu thuflc lodi Acetobacter diazotrophicus.
VI s i n h v^t p h d n gidi phot phdt k h d tan .
•Phltphorlt Id mgl nguyfin t l dinh dirdng quan ti-ong cho cdy. Mgt s6 VSV cfl t h l be gay nhung iifin k i t p h i t phdt, c h d n g phan gidi nhung lifin k i t phltphoril vd chuyen thdnh dgng cay trong cfl t h l s d dyng flifpc, d i n d i n Idm tdng sinh tru'dng vd nang s u i t flflng rugng. VSV co t h l cung d p khoang 20 - 2 5 % nhu c l u phltphorit cOa cay (Bajpai vd Sundara, 1971).
B i n g 4, a l lu'ong vi sinh vat phSn giai lan trong cac m l u dat (x lo'CFU/g tr9ng lu'png kho) - . M l u
S6 lupng
VSV
D i t lr6ng ci phe CFI
7.77 CF2
16,8 CF7 69.7
CF8
37,2
B i t tring ch6 j Dat trong h i tifiu ' C1
16.9 C2
49 C I I
24,5
C12 [ TCN.C
46.2 26
TCN 1 TCc
19,7 ' 27,2 G3 j HT14
8.66 34,2 HT4
27,5
HUI NUHl KHOA HoC CONb NbHb hINH HOC TOAN QUOC 2013
K i t qua phan ttch nigt flg
^^r
~ V S V phdn gidi ian trong
cdc miu dat b Bdng 4 vd
Hinh 4 cho t h i y cdc mau cd s l l u p n g khodng 10^CFU/g a l t kho. Ve s l
" l u p n g h h 5 m ' V S V ndy so vdi d i t t r i n g n l i chung la
khflng t h i p do VSV phan „ „ ^^^ CFS g i i i ldn cfl kha nang s i n g . , ,
frong d i l u kign nghfio dinh " ' " ' ^ *• ^ = ' " ' ' ^ * P ^ ^ " S ' ^ ' ' ^ " •^""9 '^^^ " ' ^ ^ ^ ^ ' •^""9 " l " " ^ 9 " ^ ^ "
dirflng, khi flfl chiing phdn gidi cdc logi p h i l phdt kho tan thdnh n g u i n p h i t pho cho sinh t n r d n g va phat trien. Tuy nhifin hogl tlnh cOa cdc chiing VSV phdn gidi Idn quan sdt t h i y d cdc m l u a l l nay id khflng cao.
L":J
Bing S. MCFC d^ phdn gldl Caj(P04)i c
KI higu chiing
Him lupng P tan (mg/l)
ua Cac chiing vt sinh vgt d i m e t u y i n chpn VI khuin VL-CF
7.3 665
VL-CF 8.2 475 5
VL-C 12.1 533
VL-C 1.5 472 5
VL-HT 4 2 500
VL-HT 14.5 59 25
Nam moc ML-CF
1.3 8 5
ML-C 2 6 63 5
ML-HT 14 2 591,25 T d 14 m i u d i t chiing 161 fld hjyfin
chon flupc 6 chOng VK vd 3 chiing n i m m I c cfl khd ndng phan giai lan cao tren mfli trudng chda n g u i n p h i t phdt k h d tan (Bdng 5).
Chiing VSV cfl khd nang phdn gidi h p p - c h i t photpho'cao c^ira han Id cfl dnh hudng tot d i n cdy t r i n g . Vi ngodi hogt tlnh phdn gidi Idn, n h i l u ChCing VSV efln cfl cdc hogt Unh sinh h(?c khdc gay anh h u d n g x l u
d i n sinh trudng, phat trien v d ndng s u i t cay trong. Do vay sau khi danh gid khd nang phdn gidi ldn, cdc chung V S V diing d l sdn xudt phdn vi sinh d n flupc phdn logi vd fldnh gid dnh h u d n g flen doi t u p n g cdy trong s d dung.
3 chiing VK vd 3 chung n i m m I c cfl hogt tinh cao thich dng v d i sinh li cua tdng logi cdy d u p c chiing tfli luyen d i g n . Dya trfin cdc age flllm hinh thdi, sinh II. sinh hod 1 «hung VK flupc xac djnh Id Badllus subtilis VL-CF7.3,1 chiing VK Id Bacillus megatherium VL-C12.5 vd 1 chung VK Id Pseudomonas putida VL-HT14.5. 3 chOng n i m moc co khd ndng phdn gidi Idn d l u fliugc nhflm Aspergillus. Cdc VSV ndy fleu Id nhdng VSV ao todn cho ngudi vd v g l nufli cdy trlng.-^ • Ddnh g i a kha ndng s i n h c h i t kich t h i c t i s i n h t r u d n g A I A
T i l e d cdc chiing t u y i n chgn flupc nufii trfin mfli t r u d n g cfl b l sung tryptophan vd xac djnh khd nang sinh AIA. Sau khi b l sung t h u l c tiid SalkowsW vd so mdu thu flupc 3 chiing Azotobacter. 1 rfiflng Acetobacter vd 1 chung V K phdn giai ldn cfl khd nang sinh AIA. Hdm l u p n g AIA cao nh.it thu d u p c t u A. c/injococum Ab-C11-.1 (28,2 - 12,4 mg/l) sau dfl Id A. chroococum Ab-CF7.2 (25,2 - 1 4 , 4 mg/l), A. vinelandii Ab-HT14.2 (21.5 - 1 4 , 8 mg/l), Acetobacter diazotrophicus Ac- - CF2.2 (20,5 - 1 3 , 6 mg/l) va c u l i cung Id P. puUda VL-HT14.5 (18.5 - 1 2 , 5 mg/l) (Hinh 6).
Nghifin c d u t g o - c h l p h i m v i s i n h v | t c h i > c n d n g c h o cay c h e , ca p h f i i h o t i f i u - ' """ " " "' " ' ' - ' • • - • Cdc chiing VSV flupc tuyen chpn thlch
dng -cho m i i logi c&y tiong flupc t i i n hdnh nuoi c i y trfin mfli ti-udng x6p thanh hdng. Chiing I6i dd xdc fljnh kha ndng cOng t i n tgi tifin mfli t r u d n g x I p ciia cdc_chung VSV chdc nang trong cdc tgp hpp c h i n g . Nghifin c d u ndy r i t c i n rfio vigc l y a chgn cdc t l h p p chiing trong san x u l l phan vi sinh-fla chiing, da chdc ndng. Cac cflng tiinh cflng b l cho t h i y s v phi^' hpp gida cdc chOng VSV Uiudng cho higu qud t i t h o n khi dOng Chung dcrn 11. Tuy nhifin khflng
phdi bg chOng VSV co (ch ndo khi p h l i hpp vdi nhau cijng cho felt qua tot. V i vgy phai kiem ti'a anh h u d n g t u c n g lac 'gida cdo chung khi phoi hpp v d i nhau trong c h i p h i m vd sau flo Ia"anh h u d n g Ifin cdy trong (Hinh 7).
Trang cdc Iogi c h l p h i m VSV, chung g i l n g cfl 9 nghTa q u y l t fljnh a l l vdi chat l u p n g ciia san pham. Tuy nhifin ngodi hogl Unh sinh hgc cao, cdc chung phdi cfl khd nang cgnh tianh de sinh tiudng vd hoat dong tot t r t n g cdc mflt t r u d n g sinh thdi d i t . D i n g thfli cdc chung phdi g i u d u p c hogl tinh qua thdi gian bdo qudn thi mdi dam bao cfl qur trinh cflng nghg sdn x u i t I n fljnh. Cdo c h l p h i m flupc toi u u hoa t h ^ h p h l n va flilu kign nufli c l y d l dgt mgt dg VSV h u u I'ch I d 10* CFU/g t r d ldn sau 1 h j I n c l y vdo vd fld flupc ttieo dfli b i l n agng mdt ag VSV tiieo thdi gian bdo quan d nhift flg phflng. Khi nghifin c d u khd ndng t i n tgi cua cac chiing VSV tnang c h l phdm chiing tfli nhgn thay chiing fliu sinh frirdng vd phdt b-iln t i l trong c h l p h i m . C h l p h i m cfl t h i bdo qudn tiring 6 thang d nhigt dg phflng (dgt m § l flg tiieo tifiu c h u i n ngdnh Nflng nghifip TCN 6167-1997). Hogl tinh sinh hpc ciia cac chiing cGng khflng bj mat di sau thdi gian bdo qudn.
r
Hinh 7. Thi> nghiem c h l p h i m vi sinli chdc nang len che, ca phe, ho lieu
HOI NGH! KHOA HQC C O N G NGHg SINH H p C T O A N Q U O G 2013
Tren c o s d qui tiinh chung de san x u i t chfi pham VSV cOa Phflng Cflng nghg vSt ligu a n h hpc. Vign Cong nghg sinh hgc, chiing 111 da nghifin cdu cac d i l u kien Uiich d n g v d i cdc chiing VSV d u p c tuyen chpn fli xay d y n g qui ti-lnh s a n x u i t che p h i m VSV da chuc nang cho cay chg, ca phfi, ho tifiu.
K t T L U i f l i N
S l lupng VK h i i u kh!, n i m m I c va xg khuan long s l ciia 14 m l u flat ti-Ing ch6, cd phe. h i Ugu d Tay Nguygn d m d c tiung binh.%dt dp cdc VSV cfl ich cho cay fring t h i p h o n nhieu so v d i cdc Iogi flat nflng ngNgp M i d c Trong cdc m l u flat d u p c k i l m tra, 3 m l u cfl s y ton ^ i ciia nhflm VJ< co djnh nito' Azotobacter, 11 m i u cfl s y ton tgi cOa nhdm VK c 6 fljnh nitc Azospirillum. 8 m l u cfl s u t i n tai cOa nhflm VK c l fljnh nitc Acetobacter. V S V phdn gial p h i l phdt khfl tan c o mgl trong l i t ca cdc mau flat
9 chiSng VK thugc cdc nhflm Azotodacfer, Azospirillum vd Acetobacter cd kha nang d ^ n h nita t i t n h i t ; 3 chung .VK v d 3 Chung vi n i m cfl khd ndng phan giai Idn tot n h l l d u p c t u y i n . c h g n . Trong s l cdc chdng nd'y c 6 3 chiing V K A z o t o b a c t e / f l chQng VK A c e t o t a r t e r vd 1 chOng V K phdn giai ldn cfl kha nang smh chat kIch tiiich sinh t r u d n g AIA.
Cdc chflng d u p c fldnh gid Unh I n djnh hoat Unh cung n h u kha ndng cOng t i n tgi a l tgo c h l p h i m vi sinh da chdc nang thich dng cho tdng logi cdy chfi, cd p h i , h i U l u . Cdc c h l p h i m d u p c k i l m tra deu dgt mgt dg theo ti§u c h u i n phan bfln Vigt Nam.
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Soil Sci Soc Arn Pmc. 29 677-678.
EVALUATION OF MICROORGANISMS CONTENT IN THE PLANTING SOIL OF TEA, COFFEE, BLACK PEPPER IN TAY NGUYEN AND SELECTION OF USEFUL MICROBIAL COMBINATIONS TO INVESTIGATE MULTIFUNTIONAL
BIOFERTILIZER FOR SUSTAINABLE DEVELOPMENT CROPS
Nguyen T h l T h u \ Nguyen Van T h a o \ D o Thl G a m \ Tran Dinh Man*^, Nguyen The T r a n g ^ P h a m T h a n h H a ^ Tran T h i Hoa^"
' c e n t e r for high technology development, Vietnam Academy of Science and Technology Institute of Biotechnology, Vietnam Academy of Science and Technology
SUMMARY
In ihe framework of Ihe TN3/C01 projecl, we have investigated theaetobic fkiiaofsome plating areas oftea,co&e, black pepper in Tay Nguyen.
The number of total aerobic raicroorganisms reached at medium levels In the soils the numbers can exceed ~ 10' CFU/g (dry weight) for fimgi, -• lO' CFU/g fbc.actinomyces and - 10' CFU/g for aerobic bacteria. However, the density of beneficial raicimiiganisnis to planls is much lower than that in other types of agricultural soil. Some of microorganism strains showing flie nitrogen-fixing activity, the phosphate solubiliang acdvi^ and synthesizing a plant growtfi promonng factor such asindole-acetic acid have been isoUted, selected from these soil samples In total of 14 soil tested samples. * e group of free nitrogen-fl^ing bacteria Azolobacier was present in 3 samples, flie group of nitrogen-Gxing bacteria Azospinllum was observed in 11 samples. Ihe group of mtrogcn-fixing bacteria ^cew6oc(er was obtained in g samples and the phosphate soluhilizutg microorganisms wei« presented in all samples. The selected strains were tested for some biological activities and preliramarily classified. ID fliis collection, 9 strains o f high tiitrogen fixauon bacteria have been selected, mcluding 3 strains belonged lo the getuis Azolobacter, 3 strains identified to Azospirillum genus and the rest was classifed in Aceiobacirr genus. Moieover, 3 bacterial strains and 3 firagal strains showing h i ^ phosphate solidiilizing activity were selected- Among the selected strains that were capable of fixing nitrogen or phosphate solubilizing. 5 strains inciuding 3 strains of Azotobacter, 1 strain of Acetobacter and 1 strain of phosphate solubilizing bacteria were able to produce AIA. which is a plant growth prrawting factor. Finally, the compenuon of Ihe strains present ta combination was evaluated to develop 3 types of multifunctional biofeiraizer for each type of tea, coffee and black pcppCT planls. Three lypes of multifiincEioQal biofettilizeis were produced and used for these crops.
Keyvwds: black pepper, coffee, nitrogen-fixing bacteria, phosphate solubilizing microcHganisms, synthesiring indole-acetic acid, tea.
Author for correspondence: Tel: + 84-4-37567103; E-mail: MmaniSubt.ac.vn