Tap chi sd 14, thang 6-2015, trudng Dai hpe Tay Nguyen ISSN I859-461I PHAN LAP, TUYEN CHON CAC C H U N G VI KHUAN C O DINH N I T O AZOSPIRILLUMLIPOFERUM T R O N G R £ CAY N G O TAI T I N H DAK N O N G
Nguyin Tiin Dung', Tran Minh Dinh S Nguyen Anh Diing' Ngay nhan bai: 01/6/2015; Ngay phan bien tiidng qua: 12/6/2015; Ngay duyet dang: 15/6/2015
T O M T A T
Nghien ciru tien hanh phan lap va tuyen chpn cac chiing vi khudn cd dinh N Azospirillum sp npi sinb trong re cay ngd duac thuc hiSn tai mpt sd huyen cua tinh Dak Ndng. Kit qua la 43 chiing vi khudn da dupc phan lap tir rl cay ngd. Sii dung ky thuat PCR vdi cap mdi dac hieu tiiiit ki dua tren ttinh tu gen nifH cua vi khuan ^zoj/)/r///um//>o/eram da dinh danh dupc 04 chung la K17,K25,K55vaD06. Dac diem sinh bpc ciia cac cbung tuyen chpn dupc md ta va xac dinh hoat tinb sinb tong hop lAA. Kit qua cho thay kha uang tdng hop lAA cua 4 chung tir 8,86-14,60 mg/L. Thii nghi§m dupc tiin banh tren ddng rupng bang each gay nhiem 4 chiing tren hat giong va bdn N giam 0, 25, 50 vd 75% theo qui ft-inh. Kit qua cho thay cac chi tieu smh tnrdng nhu chieu cao cay, sd la, ham lupng diep luc trong Id va nang suat d cac cdng thiic cd cbung vi khuan cd dinh N Azospirillum K17, K25, D06 va giam 50% N khdng khac biet cd y ngbia thdng ke vdi cdng thiic khdng chung vi khuan va bdn 100% N so vdi qui ttinh. Cdng thiic chimg vi khuan K17 bdn 75% N theo qui trmh cd nang suat 108 ta/ha cao ban 20% so vdi cdng thiic khdng chung vi khuan va bdn 100% N so vdi qui tiinh.
Tvr khoa: Azospirillum lipoferum, co dinh N, cdy ngo, tong hgp lAA.
1. M O DAU
Tay Nguyen la mdt vung cd tiem nang ddi dao de phat trien ndng lam nghiep va pbat tnen cac vung chuyen canh san xuat hdng hoa. Tdy Nguyen cd dien tich dat ndng nghiep gan 2 trieu ha, bao gdm cac tinh Kon Tum, Gia Lai, Dak Lak, Dak Ndng va Lam Ddng. Ngd ciing la mdt ttong nhiing cdy ttdng chu luc ciia Tay Nguyen, vdi dien tich gan 250.000 ha, chiem tten 20% dien tich ngd cua ca nudc. Ddk Ndng Id tinh cd dien tich ngd khoang 35.000 ha, nhung cd su da dang ve gidng ngd ban dia cua ddng bdo ddn tdc thiiu sd. Day Id ngudn gen qui khdng cbi ttong bao tdn, chpn gidng ngd ma cdn doi vdi cac vi sinh vat ndi sinh ttong re cay ngd.
Azospirillum Id vi khuin sdng trong ri cac loai cay ngii cdc nhu lua, ngd va mdt sd Ioai cd thupc hp hda thdo. Nhdm vi khuan nay cd kha nang cd dint N, tong hap nhiiu chat kich thich sinh tnidng thuc vat lAA, GAj pban gidi phosphat khd tan,...
gdp phdn nang cao dp phi nhieu cua dat, kicb thich tang trudng, tang nang suat cay ttdng, gidm bdn phan bda hpe va la ca.sd de phat ttien n6n ndng nghiep sinh thai ben vung. Azospirillum sdng ndi sinh ttong re cay, nhung khdng cd sir canh tranh ngudn carbon nhu vi khuan khu tni tten be mat re va cd thi cung cip dam cho cdy ttdng ma khdng
phai nhd den khi te bdo chet (Akban et al., 2007;
Bashan et al, 1997; El-Komy, 2005; Enrique, 1982; Fulchieri etal., 1994).
O Viet Nam da cd mpt sd nghien ciiu, phan lap cdc chung vi khuin thudc chi Azospirillum ttong re liia, ngo (Nguyen Ngpe Diing & cs 1999;
Hodng Qudc Khanh & cs., 1999). Tuy nhien, nhiing nghien ciiu ve vi khuan cd dinh dam Azospirillum ttong ri cay ngd cdn rit it. Dac biet, hien nay cbua cd mdt nghien ciiu nao ve thanb phan loai cua chi Azospirillum trong ri cay ngd tai mpt sd dia phuang cua tinh Dak Ndng. Muc tieu cua ngbien ciiu la tuyen chpn dupc mpt sd chiing Azospirillum lipoferum cd hoat tinh cao ndi sinh trong re cay ngd d tinh Ddk Ndng.
2. VAT LIEU VA PHlTONG P H A P NGHIEN CLTU
2.1. Vgt Ueu
Cac chung vi khudn npi sinh cd dinh dam Azospirillum dupc phan lap tii ri cay ngd trong giai doan trd cd tai huyen Krdng Nd, huyen Cu Jut va huyen Dak Mil, tinh Dak Ndng. Gidng ngd Iai C919 (Cdng ty Monsanto Viet Nam) dimg trong thi nghiem ddng rudng. Vi khuan Azospirillum lipoferum ALIO (chiing duang) do Vien Nghien cim vd Phat tnin Cdng nghe Sinh bpc, trudng Dai bpc Can Tha cung cip.
1 NCS lgi trucmg Dgt hoc Tdy Nguyen
2 ThS.. Vi^n Cdng nghe Sinh hpe & Moi trucmg. trudng Dai hoc Tdy Nguyen 3 PGS. TS., Vi?n Cdng nghe Sinh hoc & Moi trucmg. truong Dai hoc Tdy Nguyen
Tdc gid lien he- Nguyen Anh DUng: DT: 0905426524; Email: nadzungtaynguyenuni(^yahoo com vn
Tap chi sd 14, thdng 6-2015, trudng Dai hpe Tay Nguyen ISSN 1859^611 Cap mdi chuyen biet de nhdn dien Azospirillum
lipoferum dupc Vien nghien ciiu vd phat ttien Cong nghe Sinh hpe, Trudng Dai hpe Can Tha thiit ki dua theo trinb tu gen nifU cua vi khuan Azospirillum lipoferum.
+ Moi xudi: 5'-GTAAAT CCA CCA CCT CCC -3'va
+ Moi ngupe: 5' -TGT AGA TIT CCT GGG CCT-3'.
2.2. Hda chdt: Acid Malic, KH/O^, Na^MoO^, CuSO,, ZnSO^, Saccarose, Agar, Biotin, Pyridoxine Id cdc hda chat tinh khiet.
2.3. Phuffngphdp nghiin cuu 2.3.1. Phuang phdp thu mdu
Ri cay ngd dang ttong giai doan ttd cd thudc nhiiu gidng khac nhau dupc thu thap mdt each ngdu nhiSn, mdi mdng Idy nam cay (4 cay d 4 gdc va 1 cay d giiia rudng) tai cac huyen Krdng Nd, Cu Jut, Dak Mil thudc tinh Dik Ndng. Todn bp bp re dupc dung ttong tiii nilon da khii tning. Cac mau sau khi liy dupc ghi nhan nai liy, ngay lay va ngudi thu mau. Man re cay ngd sau khi thu dupc dem tdi phdng thi nghiem, riia that sach dat bam d re dudi vdi nudc chay, de rao tu nhien, de rieng timg mau rdi tien hdnh phan lap hoac bpc rieng timg mau ttong tlii nilon da khir triing budc chat vd bao quan ttong tli lanh (5°C) neu chira phan lap ngay.
2.3.2. Phuong phap phdn lap
Re cay ngd dupc khii tning bang dung dich chloromine 1% ttong 30 phiit, tiep tuc khir trimg bing cdn 70" 5 phut, H^Oj 3 pbiit rdi nia that sach bang nudc cat vd trimg (6 lan). Sau dd lay khoang 2g mlu cho vao cdi gia nhuyin rdi them 0,5 - 2ml nudc cat vd trimg vdo cdi, de yen 10 phiit va hiit liy phan ttong cbo vao eppendorf, de yen 10 phiit cho dich mdu ling xudng vd lay pban nudc ttong d£ tiin hdnh phan lap. Lay 0,5ml dich nghiin re cho vdo dng nghiem chira 5ml mdi trudng NFb ban dac khdng dam (Akbari et al., 2007; Falik et al., 1996). U d nhiet do 32''C ttong vdng 4 - 5 ngay, mdi nghiem thirc lap lai 3 lin. Quan sat nhiing dng nghiem cd vdng ddm (vdng pellicle), each bi mat"
mdi tnidng 2 - 5 mm cbi tbi cd su hien dien ciia vi khudn Azospirillum. Chuyen vi khudn tii mdi tnidng ban dac sang mdi trudng dac tiin hanh lam thuan gidng.
Mo ta cac dac diim hinh thai ciia cac chimg dudi kinh hien vi, xac dinh dac diim sinh hpe va dinh danh theo khoa phan loai ciia Beigey 1994.
2.3.3. Phuang phdp xdc dinh khd ndng tgo lAA cda cac chung Azospirillum
Danh gia kha nang tao lAA cua cac chiing
Azospirillum dira tren ndng dp lAA cd ttong dich chiet nudi cay vi khudn theo phuang phap Sawar
& Kiemer vdi thudc thir Salkowskd (Sawar &
Kremer, 1995).
2.3.4. Phuang phdp dinh danh cdc chung Azospirillum bdng ky thugt PCR
Nudi cay vi khuan d pba log (sau 36''), Iy trich DNA vi kbudn va tien hanh nban DNA vdi cap mdi chuyen biet bang may PCR rdi chay dien di san pham. Dp Idn ciia doan khuyech dai Nif H la 400 bp. Phan ling khuich dai DNA dupc tbuc hien ttong the tich 25 |il. Mot phan iing bao gdm: l,0fiL DNA vi khuin; 2,5|.iL lOX PCR buffer; l,OfiL mdi xudi; l,0|aL mdi nguac; l,OjiL dNTPs (dATP, dCTP, dGTP, dTTP); 0,5jiL Taq polymerase, 18pL nudc cat vd triing.
2.3.5. Phucmg phdp ddnh gid khd ndng c6 dinh dgm cua cac chung vi khudn Azospirillum duoc luyin chon tren dong ruong
Tbi nghiem dupc thyc bien tai Trai thuc nghiem Ndng lam nghiep, Khoa Ndng Lam, Dai hpe Tay Nguy8n nam 2011. Hat ngd lai C919 dupc khir trimg bing H^O^ 3% ttong 3 phut, nia lai 4 lan vdi nudc cit. Sau dd, cho bat vao dia petri cd bdng da dupc khir trung, vdi mpt it nudc va li cho tdi khi hat nirt mdng chuan bi nay mdm.
Riia sach hat bdng nudc cit. Cac chiing vi khuan Azospirillum da dupc tuyen chpn dupc nuoi ttong dng nghiem sau 72 gid lay dich sinh kbdi te bao cho hat ngd vao va ngam 15 phiit. Vdi ddi chiing thi ngam bang nudc cat, sau dd dem gieo, moi hdc gieo 2 hat.
Thi nghiem dupc bo tri theo phuong phap khdi nglu nhien ddy du gdm 11 nghiem thiic, 3 Idn lap Iai. Nghiem thiic 1 (NTl): khdng hdn phan dam, khdng nhiim Azospirillum; NT2: khdng bdn phan dam, nhiim Azospirillum K17, NT3: khdng bdn phan dam, nhilm Azospirillum K25, NT4: khdng bdn phan dam, nhiem Azospirillum D06, NTS: bdn 50% phan dam, nhiemAzospirillum K17,NT6: bdn 50% phan dam, nhiem Azospirillum K25. NT7: bdn 50% phan dam, nbiem Azospirillum D06, NTS:
bdn 75% phan dam, nhiem Azospuillum K17, NT 9: bdn 75% phdn dam, nhidm Azospirillum K25, NTIO: bdn 75% pban dam, nhiim Azospirillum D06, N T l l : bdn 100% pban dam, khdng nhiim Azospirillum, dien tich mdi d thi nghi$m la 2m^
Cac bien phap ky thuat cbSm sdc ngd lai theo qui trinh ciia Cue khuyen ndng.
2.3.6 Phuang phdp xdc dinh hdm lu^mg diep luc Ham luong diep hic (chlorophyll) va carotenoid dugrc xac dinh bdng phuang phap so mau (Yoshida, S.,&Fomo, 1976).
Tap chi so 14, thang 6-2015, trudng DaJ bpc Tay Nguyen ISSN 1859^611 3, K E T QUA VA T H A O hV^
3.1 Kit qud phdn lgp vd ^nh danh vi khudn npi sinh Azospirillum lipoferum
Ket qua 43 chiing vl khuan ndi sinh re cay ngd dupc phan lap. Trong dd, huyen Krong Nd phan lap dupc 22 chung k^ bieu KOI, K03, K12, K15, K16, K17, K18, K19, K21, K22, K23, K25, K26, K33, K36, K37, K38, K39, K40, K51, K54, K55, huyen Cu Jut phdn lap dupc 10 chung ky hieu COI, C03, C05, C22, C23, C24, C31, C32, C33, C40; huyen Ddk Mil phan lap dupc 11 chung ky hieu D02, D04, D06, D07, D08, D09, DIO, D15, D17,D46,D47.
Cdc chimg vi khuin deu cd dac diim la khuin lac trdn, bdng, be mat ldi, mau sac ttang ttong, boac
tring due, bia khuan lac phang. Te bao vi khuan cd dang que, gram am. Cac chimg vi khuan pban lap deu sinh trudng trong dieu kien vi hieu khi, tao thanh mpt vdng pellicle ttang due ben dudi mdi trudng Nfb ban dac 2-5mm va lam bi^n ddi mau mdi trudng tir mdu xanh la sang mau xanh duong.
Cac dac diem nay hoan toan phu hop vdi nhiing nghien cim tmdc day ve vi khuan Azospirillum (Nguyin Ngpe Diing & cs., 1999; Hoang Qudc Khanh & cs., 1999; Bashan etal., 1997; Fulchieri etal., 1994 vd Falik etal., 1996).
Cac chung vi khuin dupc dmh danh bing sinh bpc phan tir. Ket qua cd 4 chimg xuat hien band 400 bp tuong ddng vdi chimg chiing duang la Azospirillum lipoferum ALIO, dd Id cac chiing K17, K25, K55 va D06 (Hinh 1).
Hinh 1. Ket qua difn di DNA ciia cac chiing Azospirillum Ghi chd: Gieng T: Thang 100 bp; gieng (-):
Chungdm, gieng (+): Chung duang Azospirillum lipoferum ALIO; gieng 1: Chung K17; gieng 3: Chung K25; giing 4: Chung K55 vd gieng 7: Chung D06, cdc giing con lgi khong phdi Id Azospirillum lipoferum (khong co band 400bp hogc CO band phu).
3.2 Khd ndng sinh tong hap lAA cua cdc chung Azospirillum lipoferum
Smh tdng hpp lAA duoc xem la mdt yen td Bang 1. Kha nang t6ng hop lAA cua
quan ttpng ddi vdi sinh trudng va kha nang khang ban cua cay ttdng (Akbari et al., 2007; El- komy 2005 va Naiman et al., 2009). Bdn chung Azospirillum tuyen chpn dupc danh gid boat tinh sinh tdng hpp lAA (Bdng 1). Ket qua cho thiy 4 chung tuyen chon deu cd kha nang smh tdng hpp lAA. Kha nang sinh tdng hpp lAA bien thien tii 8,62 den 14,60 mg/L, tuang duang vdi chung ddi chiing ALIO. Rieng chiing K17 cd khd nang tdng hop tdi 14,6 mg/L cao hon chimg ddi chiing va tuong tuong vdi mpt sd chung ciia nudc ngoai.
cac chung Azospirillum lipoferum
STT 1 2 3 4 5
Chiing vi khuan — K17 K25 K55 D06 ALIO LSD 0.01
C V ( % )
Lan 1 13,58 9,29 11,22 10,64 9,80
Ham lirong Lan 2
15,71 9,33 10,98 12,14 11,50
IAA(mg/l) Lan 3
14,50 7,26 10,49 9,53 11,10
Trung binh 14,60
8,62 10,90 10,77 10,65 2,35 10,26%
Tap chi sd 14, tiidng 6-2015, trudng Dai hpe Tay Nguyen ISSN 1859-4611 Kit qua nghien cim cua Cassana et al. (2009)
cbo thay Azospirillum brasilense Az39 cd kha ndng san xuit 12,73 - 13,49 ng/mL (d mat dp 3,4 X 10« t i bao/ml). Akbari et al. (2007) da phan lap vd tuyin chpn dupc mpt sd chiing Azospirillum sp cd kba nang sinh tdng hpp lAA kich thich sinh ttudng cua cay Iiia mi. Ket qua la su nhiSm cac chiing Azospirillum ssp da lam tdng dudng kinb gdc Ilia, chieu dai re, ttpng lupng khd va sd lupng long re so vdi ddi chumg.
3.3. Anh hu&ng cua cdc chung Azospirillum lipoferum din sinh tru&ng vd ndng suat cua cdy f^go ngodi ddng rupng
Cac chung Azospirillum duac nudi cay va u vdi hat gidng trudc khi gieo. Ket qua ve anh hudng cua Xli ly chung, lieu lupng bdn N den ham lupng diep Iuc trong la, mdt cbi tieu quan trpng trong tucmg quan vdi bam lupng N cay hip thu dupc ghi nhan d bang 2.
Bang 2. Anh hirdng ciia xu ly chung din hdm luong diep luc (mg/g)
NTl NT2 NT3 NT4 NT5 NT6 NT7 NTS NT9 NTIO NTll
Nghiem thirc OVK-ON K17-0N K25- ON D06- ON K17- 50%N K25 - 50%N D06 - 50%N K17- 75%N K25 - 75%N D06 - 75%N 0VK-100%N LSD 0,01
CV (%)
Chlorophyll a (mg/g) 1,686 1,917 2,306 2,242 2,651 2,716 2,354 2,863 2,420 2,641 2,642 0,752 7,35
Chlorophyll b (mg/g) 0,857 1,008 1,184 0,954 1,253 1,405 1,371 1,483 1,200 1,447 1,245 0,38 6,88
Carotenoid (mg/g) 0,813 0,843 0,741 0,726 0,989 0,978 1,074 1,024 0,877 1,057 0,875 0,213 8,56 Ket qua d cac nghiem tiiiic xir Iy hat ngd vdi
cac cbung vi khuan ttudc khi ttdng kit hop vdi bdn 50% dam va 75% dam cho bam lupng chlorophyll a, chlorophyll b va ham lupng carrotenoid ngang
Bang 3. Anh hirdng ciia nghiem thuc den sinh trirong chi€u cao cay ngo bang vdi nghiem thiic bdn 100% dam khdng nhiem vi khuan. Su khac biet cd y nghia thdng ke d miic y ngbia P< 0,01.
NTl NT2 NT3 NT4 NTS NT6 NT7 NTS NT9 NTIO NTll LSD 0,01
CV(%)
Nghiem thirc 6 tuan
OVK-ON K17- ON K25-0N D06- ON K17- 50%N K25 - 50%N D06 - 50%N IC17- 75%N K25 - 75%N D06 - 75%N 0VK-100%N
Chieu cao cay (cm) 8 tuSn
139,50 149,83 148,00 148,33 161,33 159,33 156,17 172,17 170,50 168,00 171,67
6 tu^n 168,33 175,83 175,17 174,00 193,00 190,33 186,50 203,50 201,67 197,50 202,67 15,37 0,99
Duong kinh goe (cm) 8 tuan
25,62 30,70 27,40 32,63 27,33 32,30 26,53 32,03 31,63 36,20 31,37 36,23 29,80 35,50 35,27 40,80 35,10 40,10 34,93 39,93 35,37 40,83
6,78 1,03
Tap cbi so 14, thang 6-2015, tiudng Dai hoc Tay Nguyen ISSN 1859-4611 Tong bam lupng diep luc cua cac nghiem thiic
dao ddng ttong khoang 2,543 - 4,346 mg/g. Cu thi nghiem thiic 8 (NT 8) xii Iy chung K17 bdn 75% N cd tdng ham lupng diep Iuc la 4,346 mg/g cao ban 20% so vdi nghiem tiiiic 11 (NT 11) khdng xu ly chiing va bdn 100% N tiieo qui tiinh.
Ket qua d bang 3 cho thay cac nghiem thiic cd xu ly chiing K17, K25 vd D06 kbdng bdn N, cd chieu cao va dudng kinh gdc sau 6, 8 tuan cao ban ddi chiing tu 5-7%.
Cac cbi tieu sinb trudng cua ngd sau 6 va 8 tudn cd xir Iy chiing K17 (NTS), K25 (NT9) va D06 (NTIO) vdi miic bdn N la 75% deu cao hon so vdi N T l l bdn 100% N, khdng xu ly chiing.
Nhu vay cd the ndi cac chimg Azospirillum da cd tdc ddng hieu qua din sinh trudng cua cay ngd.
Ket qua nay tuang tu vdi mdt sd cdng bd tten tbi gidi ttong nb&ig nam gan day.
Cassana et al. (2009) da nghien ciiu phdi hpp hai cbung Azospirillum brasilense Az39 va Bradyrhizobium japonicum E109 xu Ii cho ngd va dau tuang. Kit qua cho thiy xu li phdi hpp bai chung cho hieu qua cao ban so vdi xii li don chiing. Ti 16 nay mdm, chieu cao cay, chieu dai re, ttong lupng khd diu tang so vdi ddi chiing.
Do Xli l^ chiing Azospirillum cd anh budng tich cue din ham lupng diep luc ttong la, tham gia true
tiep vao qua trinb quang hop, tang sinh trudng cua cay ngd, ldm ca sd cbo gia tang nang suat (Hinh 2). 6 nhdm nghiem thiic khdng bdn N va xir ly chiing K17, K25 va D06 da tang nang suit 30-50%
so vdl ddi chiing. Ket qua hinh 2 cung cho thay xu ly chiing K17, K25 vd D06 bdn giam 50% N cho nang suat ngd khoang 80 ta/ha, khdng cd su kbac biet cd y ngbia thdng ke vdi bdn 100% N tbeo qui trinh vd kbdng xu ly chiing. Dac biet, bdn 75%
N va cd xir ly chiing deu cho nang suat cao ban tu 10-20% so vdi ddi chiing bdn (NTl 1). Ket qua hmh 2 va cac ket qua bang 2, bang 3 cung xac dinh dupc chiing K17 cd hoat tinh cao nhat ttong cac chimg dupc chpn lpc, chiing nay cd the cung cap 34,5-69,0 kg N/ha/vu. Ket qua nay tuang duang so vdi mdt sd nghien ciiu ttong vd ngoai nudc.
Fulchieri & Frion (1994) nghien ciiu su dung 3 chimg Azospirillum de xu ly hat cho ngd. Ket qua cho thay da lam tang chieu cao cay, tang sinh kbdi vd tang nang suat ngd len 59% so vdi ddi chung, cac chiing nay cd the cung cap khoang 60 kg N/ha.
Fallik & Okon (1996) ngbien ciiu sii dung Azospirillum brasilense ket hop vdi than bim cho Setaria italica vd ngo. Kit qud cho thdy chiiu dai thdn, trong lugng kho, ndng sudt tdng ddng ke.
Sinh khoi vi khudn Id 10^ CFU/lg than biin co hiiu qud gdy nhiim hgt giong cao nhdt.
NT1 NT2 NT3 NT4 isn"5 NT6 NT7 NTS NT9 NTIO NT11 Nghifm thirc
Hinh 2. Anh hudng ciia xu ly chiing va lieu lupng phan N den nang suat ngo Ghi chu: cdc cot CO chir cai khac nhau Id khac biel co y nghia thdng ki & xdc suat P<0,01
Tap chi sd 14, tiiang 6-2015, fairdng Dai hpe Tay Nguyen ISSN 1859-4611 4. KET LUAN 1^'?°B P^^^" '*?"' (siani 34,5 - 69,0 kg N/ha) ma cac Su dung ky tiiuat PCR vdi cap mdi dac hieu '^hi «eu sinh ttudng, nang suit cua cay ngd kbdng tiiiit ki dira tten ttinh tu gen nifH ciia vi khuin '^o su khac biet vm tin nghi6m bdn 100% tiieo qui Azospirillum lipoferum', chiing tdi nhan dien tnnh.Dacbiet,chung^zo.;,/W//i/™/,>o/en.mK17 duoc cbung K17, K25, K55 va D06 la vi khudn ^h^ ^et hop vai bon dam a miic 75% (103,5 kgN/
Azospirillum lipoferum. Thi nghiem ttdng ngd ha) cd cdc chi tieu nang suat tang cdy nghia tiidng ngoai ddng da cho tiiiy cac chiing vi khuin ke so vm nghiem thuc khong nbiem vi khuan va Azospirillum lipoferum cd thi tiiay tiii 25-50% bdn dam 100% dam (138 kg N/ha).
ISOLATION AND SELECTION OF NITROGEN FIXING RHIZOSPHERE
^XCI^MXPL AZOSPIRILLUM LIPOFERUM OF CORN IN DAK NONG PROVINCE
Nguyen Tien Dzung*, Tran Minh DinhS Nguyen Anh Dzung*
Received Date: 01/6/2015; Revised Date: 12/6/2015; Accepted foi Publication: 15/6/2015 SUMMARY
Isolation and selection of nittogen fixing rbizosphere bacteria of com planted Dak Nong province was investigated. There were 43 strains isolated and 04 sttains identified to be Azospirillum lipoferum K17, K25, K55 and D06 by PCR technique. lAA biosynthesis activity ofthe selected sttains were also determined and showed that the sttain were able to biosynthesize from 8,86 to 14.60 mglAA/L. Effect ofthe selected strain on growth and yield of com was conducted on the field by inoculation of com seeds and reduction of 0, 25, 50 and 75% nitrogen fertilizer usage as the technical guide. The results showed that the strains enhanced the growth, pigment content and yield ofthe com compared to the conttol. The yield of com tteated W\ih Azospirillum lipoferum K17 was 10,8 tons/ha and 20% higher than the conttol as the technical guide.
Keywords: Azospirillum lipoferum, nitrogen fixing, corn, lAA biosynthesis.
TAI LIEU THAM KHAO Tai lieu tiing Vi|t
Nguyin Ngpe Diing, Hd Thi Kim Anb, Pham Thi Hoai Anh (1999). Anh huang cda cdc chung vi khudn cd dinh nita trong ri liia lin sinh truang cua mdm lua CR203, Bao cao khoa bpc Hpi nghi CNSH toan qudc, Hd Ndi, Nha xuat ban Khoa bpc Ky thuat, 416-420.
Hodng Qudc Khanh, Nguyen Phiic Hoang, Ngo Diic Duy, Le Kim Hue (1999). San xudt axit indolaxetic bai vi khudn cdn tang Azospirillum brasiliensie vd hiiu ung gia tdng kich thuac thuc vat. Bao cao khoa hpe Hpi nghi CNSH todn qudc. Ha Npi, Nha xuit ban Khoa hoc KJ thudt, 412-415.
Tai lieu tieng Anh
Akbari, G. A., Arab, S. M., Alikhani, H. A. et al. (2007). Isolation and selection of indigenous Azospirillum spp and the lAA of superior sttains effects on wheat roots". World Journal of Agricultural Sciences, pp 523-529.
Bashan, Y. & Holgui, G. (1997). Azospirdlum-plant relationships: environmental and physiologycal advances (1990-1996). Can. J. Microbiol, 103-121.
Cassana, P., Perriga, D., Sgroya, V. et al. (2009). Azospirillum brasilense Az39 and Bradyrhizobium japonicum E109, inoculated singly or in combination, promote seed germination and early seedling growth in com (Zea mays L.) and soybean {Glycine max L.). European Journal of Soil Biology, Vol 45, pp 28-35.
Diaz-Zorita, M and Femandez-Canigia, M. V. (2009). "Field performance of a liquid formulation of 4 Doctorate student at Tay Nguyen University
5 Master. Institute of Biotechnology and Environment. Tay Nguyen University 6Dr, Institute of Biotechnology and Environment, Tay Nguyen University
Corresponding Author- Nguyen Anh Dzung; Cellphone: 0905426524; Email: [email protected] 10
Tap cbi sd 14, tiiang 6-2015, ttudng Dai hoc Tay Nguyen ISSN I859-46I1 Azospirillum brasilense on dryland wheat productivity", European Journal of Soil Biology, Vol 45, PP3-11.
El-komy (2005). Coimmobilization of Azospirillum lipoferin and Bacillus megaterium foi successfiil phosphorous and nittogen nutrition of wheat plants, FoodTechnol. Biotechnol. 43(1) 19-27.
Enrique A. Rodriguez Caceres (1982). Improved medium for isolation of Azospirillum spp". Applied and Environmental Microbiology, Vol 44, pp 990-991,
Fulchieri, M. & Frion, L (1994). Azospirillum inoculation on maize {Zea Mays): effect on yield in a field experiment in centtal Argentina", Soil BiolBiochem. Vol 26, pp 921-923.
Fallik, E & Okon, Y. (1996). Inoculants of Azospirillum brasilense: biomass production, survival and grovrth promotion of Setaria italica and Zea mays. Soil Biol Biochem, Vol 28, pp 123-126.
Naiman, A. D., Lattonico, A., Salamone, I. G. (2009). Inoculation of wheat with ,^zospin7/um brasilense and Pseudomonas fiuorescens: Impact on the production and culturable rhizosphere microflora.
European Joumal of Soil Biology, Vol 45, pp 44- 51.
Sawar, M. & Kremer, R, J (1995). Determination of bacterially derived auxin using a microplate method.
Lett. Appl. Microbiol. 20, 282-285.
Yoshida, S. & Fomo, D. (1976). Laboratory manual for physiological studies of rice. Philippin: IRRI., pp. 43-45.
Phan bien 1 Phan biSn 2 TS. Nguyen Van Nam TS. Nguyen XuSn An Kboa Ndng Lam nghiep, trudng DH Tay Nguyen Khoa Ndng Lam nghiep, tnrdng DH Tay Nguyen
Dien tiioai: 01266789268 Dien thoai: 0973162850 Email: [email protected] Email: [email protected]
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