K i t qua nghidn cu-u khoa hpc BVTV - Sei 1/2012
PHAN LAP VA TUY^N CHQN CAC
N G U 6 NVI
K H U A N D 6 I K H A N G N A MBOTRYTIS CINEREA VA COLLETOTRICHUM GLOEOSPORIOIDES GAY HAI TREN RAU QUA
Isolation and Selection Antagonistic Bacteria Against Botrytis cinerea and Colletotrichum gloeosporioides Damaging on Vegetable and Fruit Crops
VQ Duy Hidn, Mai Vdn Qudn, Ld Thj Thanh Thuy Vi$n Bdo vd thyc vdt
Abstract
Total 51 isolates of bacteria were collected from 10 plant species (Aglai, chesnut, bean, star fruit, longan, custard apple, plum, peach, Chinese cabbage and IVIexican sunflower). Six isolates VK 50. VK 37, VK 55, VK 34, VK 39, VK 53 of them are capable of inhibiting the growth of Botrytis cinerea on PDA medium. The site of the inhibiting loop gain from 4,6 to 8,1mm and three isolates VK 39, VK 53, VK 55 are capable of inhibiting the growth of Colletotrichum gloeosporioides on PDA medium. The site of the inhibiting loop gain from 4.6 to 7.1mm. The VK 53 isolate is the most capable of inhibiting the growth of Botrytis cinerea and VK 55 isolate is the most capable of inhibiting the growth of Colletotrichum gloeosporioides.
Keywords: Isolate, Bacteria, Inhibiting the growth, Botrytis cinerea, Colletotrichum gloeosporioides.
DAT VAN D^
Ld cdc lodi ky sinh gay bdnh khdng chuydn tfnh, n i m Botrytis cinerea vd Colletotrichum gloeosporioides phdt sinh vd gdy hgi trdn nhilu logi cdy trong a cdc vung khdc nhau. Nam dd du-gc phdt hidn trdn cdc cay: cd chua, 01, d i blp, xodi, vai, mdn, ddo, ddu tdy, Igc v.v. vd gdy cdc tridu chu-ng thoi hog, thii qua, chit cdy con, d i m Id hay thIi thdn, thIi r l . Cdc bidn phdp gilng chIng chju, ludn canh, bOn phdn hp-p ly, xu- ly nhidt dO. logi tn> cdc lodi cd Id ky chu trung gian vd d c thulc trCi- nIm, cdc c h l phIm
Trichoderma hazianum, cdc ddng vl khuin, nIm men dd du-p-c su- dyng de phdng tru- nIm S.
cinerea, Colletotrichum spp. (Tronsino, Ystaas 1980; Agrios 1988; Bailey, Jegger 1992; Ddng VQ Thj Thanh, Hd Minh Trung 1995).
Trong nhO-ng ndm g i n ddy nhim tim ra cdc logi thuoc bao vd thyc vdt an todn v6i con ngyO-i vd thdn thidn vO-i mOi tru-O'ng cdc nghidn CCPU Q-ng dyng cdc san phIm ty nhidn d l phdng trCr sdu bdnh hgi cdy trIng dd du-p-c nghidn cil-u 0- nhilu nyO-c trdn t h l gidi. MOt s l sdn phIm sinh hpc tru- bdnh cdy dd dyp-c thyang mgi hOa nhy Aspire {Candida oleophila 1-182) trO cdc bdnh do nIm Botrytis spp., Penicillium spp. Bio-save 10LP, 110 {Pseudomonas syringae) tru- d c bdnh gdy ra bl-i cdc nIm Botrytis cinerea. Penicillium spp., Mucor pyroformis, Geotrichum candidum (Brian B.
McSpadden Gardener, Deborah R. Fravel, 2002).
Tgi Vidt Ngm 10 dOng vi khuIn phdn Idp tO- vung rd dd dypfc nghidn cu-u, su- dyng d l phOng
trO- nIm C. lagenarium gdy bdnh thdn thu- trdn cdy du-a h l u , ddng vi khuan Pseudomonas sp. co khd ndng cao trong phdng tri> bdnh (Trin Bgch Lan vd cs, 2011). Cdc ddng vi khuan Burkholderia cepacia, Bacillus sp. TG19, Bacillus sp. B4-2 dd du-p^c ddnh gid khd ndng trir nam Colletotrichum spp. gdy bdnh trdn cdy mai vdng trong phdng thi nghidm, dOng vi khuan Bacillus sp. TGI9 cO khd ndng cao nhIt u-c c h l nIm Colletotrichum sp. phdt triln trdn mOi tcut/nq (Phgm Ngpc Thu vd cs., 2011).
Trong khudn k h i bdi bdo ndy chung toi trinh bdy nhO-ng k i t qud bu-^c d I u nghidn ci>u, tuy^n chpn d c nguIn vi khuIn c6 khd ndng d l i khdng nIm 6. cinerea vd C. gloeosporiodes trong dieu kidn phdng thf nghi$m.
VAT LI$U VA PHU'OT^G P H A P N G H I E N ClJU 1.V$tli$u
Cdc nguIn vi khuIn phdn Idp du-p-c tCr Id cdc lodi cdy khdc nhau thu thdp tgi Hd NOi, Ldo Cai.
NguIn nIm 6. cinerea phdn Idp tir d chua.
ddu tdy bj bdnh thIi xdm, nguIn nIm C.
gloeosporioides phdn Idp tCr 6t bj bdnh thdn thy.
Cdc logi mOI tryO-ng PDA. TSA, KB, NA, dung djch PBS (Phosphate buffered saline pH 7.2), Cyclohexamide...
2. Phvang phdp
Thu thdp Id vd qua cua d c cdy dn qua, d y canh, cdy thyc phIm vd cdy dgi a cdc vung khdc nhau, bao quan glQ- cho nnlu Id tyoi, khdng bj ddp nat
K i t qua nghien cu-u khoa hpc B V T V - S o 1/2012
Nghien nho lOg Id trong 20 ml PBS trong 3 phut, pha loang a cdc ning dO 10\ 10^ 10^ 10"
10^ Cy moi nIng dO tiln hdnh lly 0,2 ml djch trang deu trdn cdc dTa mOi tryO-ng KB, TSA, TSA (djch du-p-c u a nhidt dO 85°C trong 10 phOt).
Kiem tra cdc dTa sau 24, 48, 72 gid- sau d y , lya chpn cdc khuIn Igc thuin, d y chuyin vdo mdi tru-o-ng dinh du-d-ng (NA) d l su- dyng.
Ddnh gid khd ndng doi khdng cua cdc nguIn vi khuan vai nam B. cinerea vd C. gloeosporioides theo phu-ang phdp cua Stirling A.M.ef a/., 1995.
Nam S. cinerea vd C. gloeosporioides thf nghidm du-p-c nudi nhdn trdn mdi tru-O-ng PDA mOt tuan tru-O-c khi thf nghidm. Cdc nguIn vi sinh vdt du-p'c nudi nhdn trdn mOi tru-dng NA 2 ngdy tru-ac khi thi nghidm.
Tan nIm 6. cinerea vd C. gloeosporioides du-p'c dye thdnh khoanh c6 dyd'ng kinh 5mm, d y khoanh nIm vdo tdm dTa mdi tru-dng PDA.
Dung que d y dIu trdn nhung vdo cdc nguIn vi khuan d y vdo mdp dTa mdi tru-dng (cdch mdp ngodi cua dTa khoang 5 mm) theo 4 dilm doi XLPng nhau. D l i chO-ng ldm tu-ang ty vd thay cdc nguon vi khuan bIng nu-d-c d t . MOi nguIn vi khuan du-gc nhlc Igi 3 Ian (3 dTa petri).
Thf nghidm ddnh gid kha ndng d l i khdng cua vi khuIn du-p-c tiln hdnh d nhidt dO 28°C. Kha ndng doi khdng cua d c nguon vi khuan du-pc ddnh gid qua vide hinh thdnh vong Q-c c h l (vdng v6 khuan) sau 24, 48 vd 72 gid. Do bdn kfnh cua
vOng O-c c h l , dya vdo kfch thu-dc cua vOng O-c c h l d l ddnh gid khd ndng d l i khdng cua chQng.
K E T QUA V A T H A O LUAN
TO- cdc mSu Id cua 10 logi cdy (hoa ngdu, hgt dd, ddu cd ve, khi, nhdn, na, mdn, ddo, d i trIng, cue dd quy) thu thdp dyp-c tgi cdc vydn khdng su- dyng thulc bdo vd thyc vdt d Ha NOi vd Ldo Cai, Didn Bidn dd phdn Idp dyp-c 51 nguIn vi khuIn trdn cdc mdi tru-dng KB, TSA. Hlu hit cdc nguIn vi khuIn dIu cO khuIn Igc trOn, nhin bOng, mOt s l nguIn mdp cO rdng cya, khuIn Igc cO mdu trIng, trdng sQ-a, vdng hay hai hing.
Tilp tyc nudi d y vd ldm thuIn cdc nguIn vi khuIn sau dO nudi d y trdn mdi tru-dng NA, sau 2 ngdy tilp tyc ddnh gid khd ndng d l i khdng cua cdc nguIn vi khuIn ndy vdi cdc nIm S. cinerea vd C. gloeosporiodes trdn mOi trydng PDA. Kit qud thf nghidm thu du-p-c cho thIy trong 51 nguIn vi khuIn du-p-c nghidn CCPU khd ndng doi khdng vdi nIm S. cinerea vd C. gloeosporiodes chl cO 6 nguIn vi khuIn mang ky hidu VK 50, VK 37, VK 55, VK 34, VK 39, VK 53 Id c6 kha ndng hinh thanh du-p-c vdng O-c c h l vdi nIm trdn moi tru-dng PDA. Trong cdc nguon vi khuIn c6 kha ndng d l i khdng ndy d 6 nguon deu cO kha ndng doi khdng vdi nIm B. cinere vd chl c6 3 nguon co kha ndng d l i khdng vdi nIm C. gloeosporiodes dd Id cdc nguIn VK 39, VK 53, VK 55 (bangi).
Bang 1. Kha nang ddi khang cua cdc ngudn vi khuIn phdn l|ip du-p-c tO la cay v6-i nIm B. cinerea va nIm C. gloeosporioides (Vifn BVTV - 2010)
TT 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18
Ky hiQU nguon vi khuIn VK1 VK2 VK3 VK4 VK9 VK10 VK11 VK12 VK13 VK19 VK20 VK21 VK22 VK23 VK24 VK25 VK26 VK27
NguIn g l c Hoa ngdu Hoa ngau Hoa ngau Hoa ngau Hatde Hgtde Hgtde Hgtde Hgtde Dgu cd ve Ogu CO ve Dgu CO ve Khe Khe Kh^
Khd K h i Khe
M6i tru-d-ng phan Idp KB
TSA-85"C TSA TSA TSA-85"C TSA TSA KB KB TSA TSA TSA TSA TSA TSA-85"C TSA TSA-85"C TSA-85"C
Hogt lye doi khang ndm Botrytis
cinerea - - - - -
• -
- - - - - - - - - -
Colletotrichum gloeosporioides
- - - - - - - - - - - - '
- -
K i t qua nghidn CCPU khoa hpc
BVTV --86 1/2012
TT 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51
Ky hl$u nguIn vl khuan VK28 VK29 VK30 VK31 VK32 VK33 VK34 VK35 VK36 VK37 VK38 VK39 VK40 VK41 VK42 VK43 VK44 VK45 VK46 VK47 VK48 VK49 VK50 VK51 VK52 VK53 VK54 VK55 VK56 VK57 VK58 VK59 VK60
NguIn g l c K h i
K h i Ddu cd ve K h i Nhdn Nhdn Na Nhdn Na Nhdn Nhdn Nhan Nhdn Na Nhdn Na Na Nhdn Na Nhdn Nhdn Nhdn Nhdn Mdn Mdn Ddo Cai trdng Dao Nhdn Nhdn Cue dd quy Cue dd quy CCie dd quy
Mdi tru-dng phdn Idp KB
KB KB KB TSA TSA TSA-85"C KB TSA-85"C TSA TSA-85"C KB TSA-85"C TSA-85"C KB TSA TSA TSA TSA TSA TSA TSA KB TSA TSA TSA KB TSA TSA TSA-85"C TSA TSA TSA
Hogt lye ddi khdna ndm ^^M Botrytis
cinerea
•
• - -
• - +
- - +
- +
- - - - - - - - - - +
- -
• -
• - - - - -
ColletotrichumWM gloeosporioides
- s - - -
'
Tj
- - - - +
- - - - -
- - - - - - + + - - -
Ghi chu: + (fc ehl; - Khdng u-e chl
Cdc nguIn vi khuIn cd khd ndng dli khdng trdn dIu tin tgi trdn Id cdy, nguIn VK 34 phdn Idp dyp'c ti> Id d y na thu thdp tgi Hd NOi; cdc nguIn VK 37, VK 39, VK 50 phdn Idp tCf Id cdy nhdn thu thdp tgi Hd NOi; nguIn VK 53, VK 55 phdn Idp du'p'C to Id cdy ddo thu thdp tgi Ldo Cai.
KhuIn Igc cua nguIn vi khuIn VK 34 c6 hinh trdn, mdu trIng sQ-a, bOng; khuIn Igc cua nguIn VK 37 mdu trIng sQ-a, hinh trOn, b l m$t nhdn;
khuIn Igc cua nguIn VK 39 mdu trdng sQ-a, hinh trdn, b l mdt bdng; khuIn igc cua nguIn VK 50 c6 mdu trIng dye, mdp cO hinh rdng cya; khuIn Igc cua nguIn VK 53 hinh trOn, b l mdt nhdn, mdu vdng nhgt, dfnh; khuIn Igc cua nguIn VK 55 hinh trOn, mdu vdng nhgt.
Hogt tInh dli khdng cua cdc nguIn vl khuIn vdi nIm 8. cinerea vd nIm C. gloeosporioideSfM khdc nhau, bdn kfnh cua vdng i>c ehl do cdc nguIn vi khuIn ndy tgo ra dao dOng trong khodng 3,6 din 8,4 mm dli vdi nIm 6. cinerea vd 4,5 din 7,1 mm vdi nIm C. gloeosporio^s.
NhO-na nguIn vi khuIn cd kha ndng Q>e chl cao vdi nlm B. cinerea nhu- VK39. VK53, VK55 thi dIu c6 khd ndng O-c chl vdi nlm C.
gloeosporioides. NguIn vi khuIn VK 53 cd bdn
kfnh v6ng O-c chl Id 8,4 mm vdi 6. cinerea v^
5,0 mm vdi C. gloeosporioides. Ode bidt nguIn vi khuIn VK 55 cd khd ndng CPC chl xip x? nhau vt^i cd 2 logi nlm B. cinerea vd C. gldeosporioicb^
(bdng 2).
10
K i t qua nghien cO-u khoa hpc BVTV So 1/2012
BSng 2. Khd nang ire ehl cOa ede nguIn vl khuIn td-i sy- phdt triln cua ndm B. cinerea vd ndm C. gloeosporioides (Vi$n BVTV - 2010)
TT 1 2 3 4 5 6
Ky hieu nguIn vi khuan
VK34 VK37 VK39 VK50 VK53 VK55
NguIn glc Ld cdy na - Hd NOi Ld cdy nhdn - Ha Npi Ld cdy nhdn - Hd NOi Ld cdy nhdn - Hd NOi Ld cdy ddo - Ldo Cai Ld cdy ddo - Ldo Cai
Ddc dilm hinh thdi khuIn Igc Hinh trdn, trlng sQ-a, bdng Trdng sQ-a, trdn, nhdn Trdng su-a, hinh trdn, bdng TrIng dye, m6p rdng cu-a Trdn, nhdn, vdng nhgt, dinh Trdn , mdu vdng nhgt
Bdn kfnh vdng li-c che (mm) Botrytis
cinerea 6,6 3,9 7,7 3.6 8,4 6,1
Colletotrichum gloeosporioides
4,5 - 5,0 7.1 K i t qua thf nghidm trdn da cho thIy c6 cdc vi
khuan doi khdng vdi n l m B. cinerea vd C.
gloeosporioide t i n tgi trdn Id cdy cdc na, nhdn, ddo thu thdp d cdc vung Hd NOi. Ldo Cai. K i t qua ndy cung tu-ang ty vdi cdc k i t qud cua Phgm Ngpc Thu, TrIn Thj Thanh Vdn, Ld Thanh Todn, Tran Thj Thu Thuy (2011) khi nghidn cu-u ve kha ndng d l i khdng cua 3 ddng vi khuIn d l i vdi nam Colletotrichum spp. gay bdnh trdn cdy mai vdng va k i t qua cua Tran Bach Lan, Bui Thanh Giao, Nguydn Thj Thu Nga (2011) khi nghien cu-u ve kha ndng phong tru- nlm C.
lagenarium gay bdnh thdn thu- trdn cdy du-a h l u cua cdc nguIn vi khuIn phdn Idp tu- vung rd.
K E T LUAN V A D t NGHI 1 . K l t l u $ n
Trdn Id cdc cdy na, nhdn, ddo thu thdp du-pc tgi vung Hd NOi, Ldo Cai dd phdn Idp du-p-c 6 nguIn vi khuIn (VK 50, VK 37. VK 55, VK 34, VK 39, VK 53) c6 khd ndng doi khdng vdi nlm. Ca 6 nguIn vi khuIn c6 kha ndng d l i khdng vdi n l m S. cinerea vd chi c6 3 nguon c6 kha ndng d l i khdng vdi n l m C. gloeosporioides. Cdc nguIn vi khuIn VK 39, VK 53, VK 55 c6 kha ndng d l i khdng vdi ca 2 logi nlm: B. cinerea (bdn kfnh vong CPC c h l 7,7; 8,4 vd 6,1 mm) vd C.
gloeosporioides (bdn kfnh vdng u-c c h l 4,5; 5,0 vd 7,1 mm).
2. Dd nghj
Tilp tyc tuyin chpn, ddnh gid, djnh danh vd nghidn cu-u d l su- dyng cdc nguIn vi khuan tren vdo vide phdng tru- n l m B. cinerea vd C.
gloeosporioide gay bdnh trdn rau qua.
T A I Lieu THAM K H A O
Hinh 1. Kha nang doi khang vdi nam8. cinerea cua VK 53 tren moi trudng PDA sau 10 ngay
Plant Pathology.
Harcourt Brace
Hinh 2. Kha nang doi khang vdi nam C.
gloeosporioides cua VK 55 tren moi trudng PDA sau 10 ngay
1. Agrios N.G., 1988.
Academic Press, INC, Jovanovich Pubblisher801p.
2. Bailey J. A., and M. J. Jegger., 1992.
Colletotrichum: biology, pathology and control.
CAB International, Wallingford, United Kingdom.
3. Brian B., McSpadden Gardener., Deborah R. Fravel., 2002. Biological Control of Plant Pathogens: Research, Commercialization, and Application in the USA. 3 May 2002 Plant Management Network.
4. Tran Bgch Lan, Bui Thdnh Giao, Nguydn Thj Thu Nga, 2011. Ddnh gid kha ndng gdy hgi cua n l m Colletotrichum lagenarium gdy bdnh thdn thy dya h l u vd byde dau nghien cu-u bidn phdp phOng try sinh hpc bIng vi khuIn vung FI.
K i t qua nghidn CLPU khoa hpc BVTV - S6 1/2012
H0i thdo qu6c gia b$nh h$i thi/c vdt Vl$t Nam.
NXB Ndng nghidp. Hd NOi 2011, tr. 135 -145.
5. Stirling A.M., Coates L.M., Pegg K.G and Hayv\/ard A.C., 1995. Isolation and selection of bacteria and yeasts antagonistic to preharvest infection of avocado by Colletotrichum gloeosporioides. Australia Journal of Agricultural Research 46: p 985 - 995
6. Ddng VQ Thj Thanh, Hd Minh Trung, 1999.
K4t qud 6iSu tra bdnh hai trdn cdy dn qud d Vidt Nam 1997 1998, Nxb NOng nghidp. Hd NOi 1999, tr. 82-157
7. Phgm Ngpc Thu, TrIn Thj Thanh Vdn, Ld Thanh Todn, TrIn Thj Thu Thuy, 2011. Ddnh gid khd ndng gdy hgi cua n l m Colletotrichum spp.
trdn mai vdng {Ochna Integrrlma Lour)Merr. vd hidu qud cua vi khuIn d l i khdng, thulc hod hpc d l i vdi n l m gdy bdnh thdn thy. HOi thdo qu6c gia bdnh hdi thi/c vdt Vidt Nam. Nxb NOng nghidp, Hd NOi 2011, tr 146-152.
8. Tronsino A., Ystaas J., 1980. Biological control of Botrytis cinerea on apple. Plant D/s.64 1009.
Phdn bidn: TS. 0$ng VO Thj Thanh
O I ^ U T R A H I $ N T R A N G C A N H T A C , S O * O g N G P H A N B 6 N , T H U 6 C H O A H p C V A T A C D O N G C O A C H U N G D ^ N B $ N H D A O 6 N T R E N M O T S 6 G I 6 N G LUA
C H U L l / C <^ THLFA T H I ^ N H U ^
Survey on the Current Cultivation Status, the Application of Fertilizer, Fungicides and Their Effect on Blast of Major Rice Cultivars In Thua Thien Hue
Du-ang Thj Thao Trang^'^, Nguyin Thj Vdn', TrIn Thj Phyang Nhi^
Tru-ang Thj BIch Phu-p-ng^'^, Nguyin Quang C a \ TrIn Thj Thu Hd^'' Abstract
Blast is one of serious obstacles for agricultural production in Vietnam as well as other countries. The emergence and development of the disease depends rapidly on some factors, such as: the influence of soil, fertilizer, chemical fungicides and characteristics of rice cultivars. The results showed that current cultivation status of the three investigated areas in both 2008-2009 and 2009-2010 Winter-Spring crops differed from each other. The use of fertilizers among the rice growing areas were different, the dosage of fertilizer used by most farmers in the areas was not following by recommendation of producer, with a tow level comparing with the recommended dosage. Significant blast took place In Quang Loi and the average yield obtained in this area reached the lowest (-2,1 quintals per 500m^). Therefore, type of soil and structure of rice cultivars, fertilizer have a significant impact to the situation of blast and yield of rice.
Beam 75WP and Fuji-one 40EC were used populariy to prevent of blast epidemic, 59.7% (2008-2009), 58,8%
(2009-2010) of Beam 75WP and 38.5% (2008-2009), 39.4% (2009-2010) of Fuji-one 40EC. They were utilized in the heavy blast epidemic areas more than others.
Assessing directly of blast on the field Illustrated that the disease appeared popularly on 13/2 and Xi23 cultivars, especially; 13/2 cultivar was a heavy susceptible to blast disease. In addition, the blast only appeared sparsely on Khang dan, HT1, N97 cultivars. Moreover, we also cornpared the blast resistance of rice cultivars between three areas through the survey. The blast resistance of flee cultivars in different areas were not the same.
Keywords: Blast, blast resistance, rice, Winter-Spring crop.
OAT VAN Oe
Bdnh dgo On do nlm Pyricularia oryzae phan b l rIt rOng vd gdy hgi nghidm trpng cho cdc vung trIng ICia(Sandhu et ai, 2003). D l i vdi hlu hit nhQ-ng didn tfch trIng IQa trdn t h l gidi, vide quan 1;^ bdnh dgo On d I u dya vdo sy du nhdp thydng 1. Vidn Tdi nguydn, Mdi tru-dng vd Cdng nghd sinh
hpc, Dgi hpc Hul
2. Tru-dng dgi hpc Khoa hpc, Dgi hpc Hul
3. Tru-dng dgi hpc Ndng Ldm, Dgi hpc Hul. Email:
dthaotrang2002@yahoo .com
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