KHANG NAM GAY BENH THAN THU

HOI NGH! KHOA HOC C O N G NGHE SINH HOC TOAN QU(iC 2013

KHA NANG 0 6 | K H A N G NAM GAY BENH THAN THU- DUaC PHAN LAP TRfiN CAY NHO NINH THUAN CCiA MOT S6 C H O N G VI KHUAN

Nguyen H u y H o f i n g ' v f i N g u y i n T h u Hien

Vl^n NghiSn cdu h$ gen, V?#n Hkn lam Khoa hgc vi Cdng ngh$ VIgt Nam

T 6 M TAT

Nho U m ^ lo^ c9y an qua co hi$u qua Idnb ti cao a tinh Ninh Tbu^n. Tuy nhiSn trong nhirng a9m gib day, di^n tich tr6ng nho glim ding kg do sau b ^ gly bai, dSc t ^ l la b ^ Mn thu gay.ra d^ lim giam 70 - 80% sfin lugag nho. B ^ tin cong vio tit c i cic phin mgng nude cda cSy, bao g^m ca than c3y, la, cu6ng la, hoa dgc t>i|t d ch^i noa vik qui mgag, lim l i ryng hing lo^t, nhieu chiim hoa bj rung hoin toan, qui son cung bi nmg, qui Idn bj thoi. Coiletotrichum gloeOsporloides l i m^t trong hai chung nam thu$c chi Colletotrichum sp. chii y ^ gay b ^ dian diu tren ciy trdng. Vdi m^c dich sijr dyng sin phim sinh h^c dl di^t txir nim b^nh, b$

chiing vi khuin Bacillus. Pseudamonas. Burkholdena... ciia Phing Vi sinh vjt dit - Vi?a Cong ngh? sinh hgc dagc sii dyng dl xic djnh khi ning d6i khing narn. B$ chimg vi khuin gom cic chiing XL6.2; D.Ng LI; DA3.1; C48; 71)67; Ps7.1. Hai c h ^ p L i dii^rc si^ d^ing trong nghiSn cuu nay li Colletotrichum gloeosporioides vi nam NTTl dupe phan %> tilr qui nho b) nhiem b ^ n W thin thtr trfin mSi tnr&ng PDA c6 bd sung khing sinh chloraiiphemcoL Ket qui thu dug^ cho diay cic chiing vi khuin dupe siJr dyng diu c6 khi nang khing nim NTTl vi Colletotrichum gloeosporipides. Diy li mOt kit qui c6 y nghia cho v i ^ san xult chl phim sinb hgc di^ tsd nim b$nb trfin c ^ oho.

Tu khoa: Colletotnchum gloeosporioides, Colletotrichum sp., Nam thin diu, Nho

MdoAu

Cfiy n h c Ifi cfiy fin qufi ^ i r g c trAng chij yfiu d h'nh Ninh Thufin da mOt th&i dugc xem Ifi cfiy trong ' s 6 m $ r , giiip n g u * i dfin nei dfiy thofit ithfii nghfio ddi v 6 i mi>c thu nhfip 100 - 300 trieu d6ng/ha/nfim (Hofing Thi Ngfit etal., 2012) nhOng nfim g i n dfiy d l f n tlch trang nho gifim dfing kk do sfiu b#nh vfi vi#c ngu-bi ndng dfin si> dung qufi nhi^u thu6c hda hQo d3n d^n cfic sfin ph^m nho khdng Oam bfio ch6t l i f g n g . M9t trong nhtrng nguyfin nhfin chlnh lfim gifim di$n tfch tr6ng nho Ifi do bfinh thfin thir gfiy ra bAi n i m Colletotrichum sp. B$nh thfin thir gfiy b$nh trfin cfiy nho idn d i u tifin d u ^ c phfit hl$n nfim 1886 & Italia vfi lan t r u y l n vfi phfit bien m^nh d Chfiu A u trong n h i l u t h i kl qua. B$nh dfi gfiy n N l u thl$t h^i t^l Nam Phi, Nam My vfi Chile trong nhOng nfim 1950 - 1951, gfiy ton t h a t d ^ n 9 0 % di^n t l c h t r 6 n g nho, nang suSt vfi sfin lircrng gifim 83 - 100% (Anderson, W a n e n 1956; Jamadar 2007). Cf Vi$t Nam, b$nh thfin t h i f trfin cfiy nho x u i t hi$n d , Ninh Thufin nfim 1999, sau dd lan sang cfic vimg t r l n g nho khfic trfin cfi nu-tirc. B$nh t i n cdng vfio l i t ca cfic phan trkn mfit d i t cCia cfiy, flic b i f t tfip trung vfio p h l n chb\, qufi non (Michael, Erincik 2008), nghlfim trpng n h l t khi qufi btrcrc vfio th61 ki thu hoach, b^nh cd t h i gfiy gifim nfing s u i t h3'i 70-80% (Hofing T h j Ngfit at al., 2012). Chinh vi v^y, vi$c du-a ra bifin phfip phdng bi> bfinh hi$u qufi vfi xfic djnh tfic nhfin gfiy b$nh Ifi rat d n t h i l t .

V i n d l d'fing quan tfim Ifi hi$n nay lfi sv* khfing thu6c cOa cfic lo^i n i m b$nh ngfiy cfing tfing. D i l u dd d i n d i n t i n h tr$ng ngU'd'i sfin x u l t khdng ngCrng gia tfing hfim l u v n g thu6c. H | u qua d l 1^1 Ifi s i / t i n du m$t hfim li/g'ng t h u l c khfi Idn trfin sfin p h i m , gfiy finh hiTd'ng d i n si>c khde con ngtpdi, d n h i l m mdi t n / d n g . T n r d c tinh trang d d , cfic bien phfip sinh h9C d u v c nghifin ciru vfi Qua vko img dgng m$t cfich r^ng rfii.

Tnsng bfii bfio nfiy, chdng tdi t i l n hfinh phfin Ifip n i m td qufi nho bj n h i l m b$nh do n i m thfin thtr vfi thi> khfi nfing khfing nfim ciJa m $ l s6 chung vi k h u i n dfi dir^rc phfin Ifip cOa phdng Vi sinh v$t dkt, V i ^ n Cdng ngh$ sinh h p c '

NGUVeN U ? U - PHU-ONG P H A P Nguyen l i $ u :

Mdi tnrdng PDA: Khoai tfiy bd vd 2 0 % ; Glucoza 2 % ; Agar 2%. Mdi t n r d n g MPA: NaCI 5 g/l; C a o thjt 3 gfl; pH 7.

ChOng n i m Colletotrichum gloeosporioldesdugc cung c^p tCi" Vi#n Bfio v | th^rc v | t . Vi§n KH Ndng nghl$p V i ^ t Nam.

Qufi nho nhiSm n i m dirpc thu thfip tir v i r d n nho t^i Ninh Thufin.

P h i r o n g phfip:

-Phin l$p nim than thir tip qui nho Ninh Thuin bl nhlim b$nh thin thw:

P h l n v d nho bj n h i l m n i m d u g c a | t bfin mdi t r u i n g PDA c d bo sung klifing sinh cloramphenicol, il 6 2 8 " C trong 5 ngfiy. D v a vfio hlnh thfii ria, lfim sgch, tfich rdi cac lo?i nfim khfic nhau.

Phuvng phip thv ddi k h f i n g nim:

Khfi nfing d l l khfing n i m cua c h l p h i m dircrc t i l n hanh theo phirtjng phfip cua Weller vfi cOng sg- cd cfii b i l n (Weller ef a'.,1988). Dfit n i m d chlnh giua dia petri, nudi tnang 2 ngfiy d l nam mgc ISn lot. D l xac dinh khfi nfing khfing n i m cua vi k h u l h . Chung vi k h u i n du'gc d y quanh nfim, cfich deu. Sau d d dTa du-gc theo doi hrng ngfiy d l quan sfit s y phfit t r i l n cua n i m vfi vi k h u i n

K ^ T Q U A V A T H A O LUAN

Phfin l^p n i m t o qufi nho b j n h l e m b f n h n i m t h f i n thu- g a - ^ / ,• : « ^ ^ f ^ ^ ^ ^ ^ ^ J i . TV i

Hlnhl. Nho Ninh T h u i n bj nhiSm b^nh t h i n

HOf NGHj KHOA HQC C O N G NGHg SINH HQC T O A N QUOC 2013

~ N h ( H f i Cfiy fin qud Ijj b ^ h thfin lliu- gfiy hgl phS b l ^ vfi n | n g ne n h l t d- t i t ca cfic vung vfi cfic nam. B f n h thfin thtr phfit tnfin manh trong dieu ki$n thW t i l t mtra nhieu, do fim cao. khi cfiy ra dpt vfi la non n h i l u , Benh lam Ifi rgng hfing logt, nhieu chum hoa bj njng hofin loan, qufi non cung bj rung, qufi Idn bi t h i l (Hinh 1).

R i f i n v d nho bj benh dirge tfich vfi dfit h-fin mdi tnrdmg PDA cd bo sung khang sinh chloramphenicol d l logi bd cfic - w ? . ^ ' i ^ " ® ^ " ^ '^^^ " " ° ' * " ^ ' * * ^^ 28''C, dfi thu dirge chiing n i m N T T l (Hlnh 2). Chung N T T l cd mfiu t r l n g , sgi dfii vfi hon bdng. Dirdi kinh h i l n vi quang h(?c s g l n i m dfii, cd vfich d gi&a. Chi Colletotrichum r l t da dgng v l tfinh thfii, vl thd, rfit khd phfin djnh loai (Crouch et al., 2009; Damm ef a/., 2009). V i vgy, d l i^nh loai chinh xfic d n phai t i l n hfinh phfin tfch trinh tuf gen 18SrRN/\.

Hinh 2. Chiing n i m NTTl du>pc phan l | p tCr nho Minh T h u j n . A: hlnh t h i i s g l n i m trfin dia m6l tnrfrng. B: Hlnh t h i i sgi nam dLFffl kfnh h i l n vl quang hipc d$ phong dfil 100X.

Khfi n f i n g k h f i n g n i m i J

Bp chflng vi k h u i n du-gc sfl- dyng trang nghidn ci>u nfiy Ifi cac chflng vl k h u i n cd khfi nfing khfing n i m Fusarium B a n g l . Danh s i c h chung si^d^ng

Phfin ioai XL62

B.Ng.LI DAa.1

SacfZ/us spja.6.2 BurMidderia cepada Pseudomonas aarugenosa

Seiratla f^ymuthla ' Pseudomonas Buorescens

Bacillus sp. TD67

oxysponjm vk Rhizoctonia solani cua phdng Vi sinh vfit Dat (Bang 1) dfi phfin Ifip tru-dc do bao g I m cfic chOng XL6.2; O.Ng.LI; DA3.1; C48; T 0 6 7 ; Ps7.1 (NguySn Ngpc -Dung e( al.. 2008, Nguyin Thi Quynh Mai ef al.. 2010).

Doi v d i thi nghiem thfl- khfi nfing khang n i m , ngofii chflng NTTl phfin l$p du-gc td qufi nho Ninh Thufin bf n h l i m ; b | n h n i m thfin thU', chflng tdi dfi t i l n hfinh thi> d i i khfing n i m Colletotrichum gloeosporioides. Dfiy Ifi mOl trang 2 loai n i m chu y l u gfiy b f n h thfin thir h-fin cfiy nho. Vi rram moc ch^m hern so vdi vi k h u i n nfin d i so sfinh vfi phfin tfch sir sinh tnrdng cua cfi n i m vfi vi k h u i n , hal chung n i m NTTl Vfi Colletotrichum gloeosporioides dugc nudi d y tnrdc 2 ngfiy. Sau khi t h i y d u v c s g sinh torcrng cfla n i m , chung vi k h u i n sfi du-gc c l y sau. Chiing n i m dirge dgt chfnh giOa dfa petri. Sau dd, 3 churg v l ' k h u i n khfic nhau du-gc c l y xung quanh vfi d c h d i u cyc n i m (dia TN). Rifing dTa d i i chi>ng ftii chl nudi n i m , khdjig c | y vi Jdiuin (dia DC) (Hlnh 3).

Thi nghifim dirge l | p Ig] 3 l l n vfi d u ^ c theo ddi tirng ngfiy. Trong 2 ngfiy d i u tien, chu-a cd s g khae bifit nhieu giu-a dTa d i i chOng, D i n ngay ttii> 3, klch Ihu^ire vdng n i m mpc phfit trien, t i l n g i n d i n vi k h u i n .

Ket q u i sau 3 ngfiy tren Hlnh 4 cho t h i y : kfch Ihircrc vdng n i m d dia d i l chi>ng khofing 3 cm trong khi d dTa thf nghifm klch thirdc vdng n i m chl khofing 2,5 cm. ChCi-ng Id, khfi nfing n i m sinh tm-dng d dTa thf nghifm bj han c h l hon so vdi d dTa d i l chiing. T i l p tyc theo ddi thi n g h i f m , ket qua v i khfi nfing khfing n i m dirge t h i h i f n rd tir ngay thi> nfim.

Hlnh 3. Scr db bo tri thi nghiem

Hinh 4. T H nghifm dti MUng n i m sau 3 hgiy nuM d y . A: Odi k h i n g n i m NTTl; B: D l l khing n i m Co/Jetotrfcftum gloeosporioides

HOI NGH! KHOA HQC C 6 N G N G H ^ SINH HQC T O A N Q U 6 C 2013

Hinh 5: Kha n i n g d6l khang nam sau 5 n g i y nu6l c l y . Aiftdi khang n i m NTTl; B: D & khang nam Colletotrichum gloeosporioides

K i t qufi Hinh 5 cho t h i y s g khfic b i f t giira dTa dot chfl-ng vfi dTa thi n g h i f m : Tren d7a d i i c h i h i g . chiing n i m N T T l v f i Coiletotrichum gloeosporioides phat trien t i t , dud-ng kinh rOng, trdn d f u ; Trfin dTa t h i n g h i f m , 2 chung n i m c d du-dng kinh n i m bd h o n han Dae b i f t h i r d n g n i m mgc b| hgn c h l khi h i r d n g y i du-dng d y vi k h u i n . 0 cfic chung vi k h u i n D N g . L I , XL6.2, Ps7.1, 7 0 6 7 vfi C48, n i m ed xu hirdng bi ngCrng phfit h-|ln vfi tgo thfinh mgt khofing cfich giira n i m va du-dng mpc cfla vi k h u i n .

Khfi nfing khfing d m cua cfic chflng vi k h u i n c d t h i gifii thieh bfing 2 hirdng: tht> n h l t , c d t h i do v i k h u i n phdt t r i i n nhanh, mgnh h o n so vCrl n i m , nfin do canh tranh dinh dird-ng, chflng n i m khdng t h i phfit t r i i n du-gc. Thi> hai, ngofii s g cgnh tranh v l dinh dirdng, cfic chflng vi k h u i n cOng cd kha nfing sinh ra m f t c h i t cd khfi nana i>c c h i s g phfit trien cda n i m Khi n i m mpc t i l n d i n g i n d i r d n g mgc cfla vi k h u i n , du^i-i tfic dyng cfla c h i t irc c h l , nam bj han c h l sinh t n r d n g hofie bj ngimg phfit t r i l n . Tuy nhien, trong qua binh phfit trien, Idifi nfing fl-c c h l cfla vi k h u i n vdi n i m cung cd thd-i gian, d l l u k i f n t i i iru khfic nhau d timg chOng vi k h u i n ( N g u y i n Thj Quynh Mai ef al., 2010). Dieu nfiy giai thfeh n g u y f h nhfin tgi sao sau 5 ngfiy thi n g h i f m , kha nfing khfing n i m cua m$.t s i chung nhir C4, C6 bj giam dl so v d i thf k i t q u f i ' d d i khang d ngfiy thu- 3 (Hinh 4b, 5b). V I n d l nfiy dira ra hu-dng nghien ciru xac djnh d l l u k i f n tdi iru cho khfi nfing d i i khang n i m N T T l vfi Colletotrichum gloeosporioides cua cac chung vi k h u i n .

B f n h n i m thfin thir do n i m Colletotrichum sp. da dirge phfit h i f n vfi trfin khfi n h i l u lofii cfiy an qufi khfic nhau nhu- xofii, dt, c f i phfi, nho, t i f u . . . (Liu ef al., 1986, N g u y i n Thanh Hfi et al.. 2 0 1 1 ; Hofing Thj Ngfit et al., 2012). tuy nhl§n cfic nghifin ci>u thirdng ch? tfip tnjng vfio tfic h^i, t r i f u chCrng cua b f n h . Hu-&ng nghidn cu-u d i f t tiir n i m gfiy b f n h n^y cdn hgn c h l , dfic b i f t , 6 V l f t Nam chira cd m$t cdng trinh nghien cu-u nao cdng bo Ve v i n d l nfiy. H i f n nay, t r f n thj t r i r d n g cd n h i i u logi t h u i c hda hoc tri b f n h do n i m thfin thu- gfiy ndn dirge fip dyng trdn n h i l u loai cfiy t r i n g khfic n h a u . Tuy nhifin n l u dung n h i i u logi t h u i c hdS hpc vdi I l i u lirgng cao trong thdi gian dai cd t h i tifiu d i f t d vi sinh vfit cd i g i trong a l t , tgo d i i u ki#n d i efic vi smh v f t c6 hgi phfit t r i i n , lam anh hird-ng x l u d i n mdi t n r d n g , gfiy ra tfic hgi nghifim trgng d i i vdi SLTC khde cua ngirdi vfi v f t nuOi. Cl m$t s i nirdc phfit t r i i n , t h u i c d i f t nfim co n g u i n g i c hda hgc bj h g n c h l h o l e d m sir dyng C h l p h i m sinh hpc d i f t n i m ed n g u i n g i c tCr vi k h u i n d i i khang c d n h i l u u-u diem ho'n so v d i v i f c dflng t h u i c hda hpc. Si> dung c b l p h i m cd n g u i n g i c ti> vi khuan d i i khfing d i d i f t n i m gay hgi trSn cfiy t r i n g se mang Igi nhOng Igi fch Ifiu dfii trong sfin x u l t ndng nghifp nhu-. lam tang nfing s u i t cua cfiy trong, giam chi phf d i u tir, thfin t h i f n vdi mdi tn/d-ng sinh thfii. khdng finh h i r d n g d i n s i i c khde cua ngu-oi vfi v f t nudi. Dfiy cOng chinh la mgc t i e u lau dfii cCia chiing tdi.

K ^ T L U i l i N

Dfi phfin Ifip d u g c chung n i m N T T l tip qufi nho tgi Ninh Thufin. Cfic chung vl khufin XL6.2, D . N g . L I , TD67, B A 3 . 1 , C48 vfi Ps7.1 d i u c d khfi nfing ire c h l khfi nang sinh t n r d n g cfla n i m Colletotrichum gloeosporioides vk N T T l sau 5 ngfiy nudi c i y .

Cong truth nay nhan dupe sv lai tr^ kinh phi tir di tdi nhdnh dp'Vifn Khoa hgc va Cdng ngh^Vi^t Nam vdi tin di tdi: Nghien cm xdy d(mg mo hinh thu nghifm vd cdc gidi phap an toan O-dnJphdkg trif sau benh cho cdy nOo Ninh Thuan.

T A I Lr$U THAM K H A O

Anderson and H. Wanen (1956) Diseases of Frait Crops. Soil Science 82(4): 345.

Crouch JA. Clario BB, While JW. Hillman BI (2009) Systematic analysis of the falcate-spored gramWcofaus CoBatotrichum and a descriptkm of six new species of the fungus fnam wami-season grasses. Mycologia 101:717-732.

Hoang Thj Ngit, Nguyin Thanh Ha. Nguyin TTiu Hfi, Pham XuSn HJi (2012) Nghifin cdu xic djnh v i phin tich da dgng di t n j y i n cua nim Cdletdnchum gkieosporioides gSy lifnh thfin thu tren nho. Tap chiNNSF7NTi2 : I M -107

HOI NGHI KHOA HQC CONG NGHE SINH HOC T O A N Q U O C 2013

—.lamarfarMM (2007) EGolQgy.epiffgnijnJflriy aprl mananprnfinf nf anlhrrgrnp'^p nf nrapiavinq DoctOr Cf philosophy In Plar\t pethotooy:

LIU X J . LI JY, Yang JT (1986) Studies on latent infection of mango in Hainan Island. Acta Phytopathalogica Sinica 16 47-51 Michael A Ellis, O. Enncik (2008) Anthracnose of Grape- The Ohio State University Extension: 3Z08

|«tauy6n Ngpc Dung, Nguygn Thj Qujnh Mai. H i Kim Anh (2008) Antifomis - C h i phim anh h^c phdng ching bfnh thii r i - lo co r l cfiy (rang do nfim Fusarium oxysponjm vS Rbizoctonia striani gay nSn. Tap dii Cong ngh$ sinh hgc 6(4A): 735-741. - . . . NguySn Thanh Ha, HoSng Thi N g ^ NguySn Thu Ha,Pham Xufin HOi (2011) Nghign cCnj flfc llnh gay bfnh n i m Coiletotrichum gloeosporioides g$y bfnh trSn cfi ph§ d miln Bio Vift Nam. Tap chi dnh hgc 33(1): 67-7a

NguySn Thj Qujnh Mai, NguySn Huy HoSng, Nguyin Ngpc Dung (2010) Anh hu*ng cOa djch Igc l l bSo tCr cfic chiing vi khuan ch<jn Igc ten sinh tnjdng ciia nam gfly bfnh cSy Fusanum oxysponim vA Rhizodonia sdad. Hgi nghi khoe hgc ky d$m 35 nim Yiin Khoa hgc vi Cdng ngh$ Vi$t Nam- Tiiu ban khoa hgc sv s6ng: 352-353.

- Than PP, Prihastuti-H. Phoulivong S. Ta^or PW. Hyde KD (2008) Chnii anthracnose disease caused by Colletotrichum species J Zhejiang Unlv Sd B 9(10): 764-778

Weller DM (1988) Biological control of soil home plant pathogens in the zhizosphere with bacteria Annu. Rev. Phytopathol2G: 349-407.

RESISTANT ABILITY TO ANTHRACNOSE FUNGUS ISOLATED FROM GRAPES OF SOME BACTERIA '

Nguyen Huy Hoang' and Nguyen Thu Hien

Institute of Genome Research, Vietnam Academy of Science and Technology SUMMARY

Grape is an economically high effective crop in Ninh TTiuan province. Currently, the vineyard area is decreased significantly due to pests, however, especially anthracrose causes a 70 - 80% decrease in grape production. Andiracnose fimgus attacks all succulent parts of plan including stems, leaves, petioles, e.specially for flower hiiiis, herrip'! which rwiilt in tftaves, blossom and yoiingfr fhiit felling down and lotting of bigger &uit. Colletotrichum gloeosporioides, one of two fiingus strains b e l o i ^ to Colletotrichum species, caused anthracnose disease, hi order to investigate an effectively fungicidal bioproduct against anthracnose, a bacterial collection including Bacillus, Pseudomonas, Bwkholderia species named as XL6.2; D.Ng.Lli p A 3 . 1 ; C4g; 11)67; Ps7.1 strains that obtained firom Soil Microorganism Lab, Institute of Biotechnology, was used to determine their fiingicidal activities. Two tiingus strains were performed as objectives studies including Colletotrichum gloeosporioides and NTTl isolated in antiiraccose - exposed grapes on the ' PDA medium supplemented with chloramphenicol. The bacterial strains showed high fiingicidal activities, therefore here is a significant approach to produce a fiingicidal bio - product

Keywords: Anthracnose, Colletotrichum gloeosporioides, Colletotrichum sp.. Grape

"Author for correspondence: Tel: 04.37918012 Fax: 04.37917039 Email. nhhoang@igr.ac.vn