Influence of bacteria on Orobanchecrenata seed bank size, incidence and Viciafaba L. performance

EURASIAN JOURNAL OF SUSTAINABLE -AMERICAN AGRICULTURE 1074-1998 0748, EISSN: -ISSN: 1995 2017, olume(11),issue(4):pages(30-39) Published Online in h p://www.aensiweb.com/AEJSA/ 93-03, Pages: 7201 yluJ) 4(11EURASIAN JOURNAL OF SUSTAINABLE AGRICULTURE. -AMERICAN Es aa M Elabaied e al, 2017 In luence o bac e ia on O obanche c ena a seed bank size, incidence and Vicia aba L. pe o mance 1Es aa M Elabaied, 2Ahmed M E Rugheim, 3Mohammed M Hassan, 3Magdoline M Ahmed, 2Mahdi A Yahia, 3Rania A Abakee , 4Rashida M A Abusin, 3Awad G Osman, 3Migdam E Abdelgani and 3Abdel- Gaba E Babike 1M.Sc. s uden , Sudan Academy o Sciences (SAS). Kha oum, Sudan 2Depa men o Landscaping and A id Land Ag icul u e, Facul y o Ag icul u e, Omdu man Islamic Uni e si y, Omdu man, Sudan. 3En i onmen , Na u al Resou ces and Dese i ica ion Resea ch Ins i u e, he Na ional Cen e o Resea ch, Kha oum, Sudan. 4Pes s and Plan Heal h, College o Ag icul u e, Bah i Uni e si y, Kha oum, Sudan Recei ed 12 Ap il 2017; Accep ed 20 July 2017; Published Online 15 Augus 2017 Add ess Fo Co espondence: Ahmed M E Rugheim, Omdu man Islamic Uni e si y, Depa men o Landscaping and A id Land Ag icul u e, Facul y o Ag icul u e, Omdu man, Sudan., E-mail: a [email protected] Eu asian Ne wo k o Scien i ic In o ma ion.-Ame icanby au ho s and 7Copy igh © 201 This wo k is licensed unde he C ea i e Commons A ibu ion In e na ional License (CC BY). h p://c ea i ecommons.o g/licenses/by/4.0/ ABSTRACT BACKGROUND: O obanche c ena a is a holopa asi ic weed ha se iously a acks legumes and depend en i ely on hei hos s o all nu i ional equi emen s. A wide a ie y o app oaches physical, cul u al, chemical and biological ha e been explo ed agains oo pa asi e. Cu en means o con olling pa asi ic weeds a e ocusing on educing soil seed bank and inhibi ing ea ly de elopmen al s ages. OBJECTIVE: Labo a o y and g eenhouse expe imen s we e conduc ed o in es iga e he e ec s o bac e ial s ains and isola es cul u es and il a es, on ea ly de elopmen al s ages o O obanche c ena a and o explo e hei po en ial in O. c ena a con ol. RESULTS: Resul s o in i o expe imen s e ealed ha ge mina ion o O. c ena a signi ican ly dec eased a e inocula ion wi h bac e ial cul u es o il a es. Bac e ial isola es (ISO43 and ISO44) and s ains o Bacillus ci culans, B. mega he ium a . phospha icum (BMP) and he combina ion o BMP plus Rhizobium leguminosa um b . iceae (TAL1399) signi ican ly educed ge mina ion as compa ed o o he mic obes and con ols. Wi h espec o he e ec s o bac e ial cul u es o il a es on haus o ium ini ia ion, esul s displayed ha B. ci culans, BMP, BMP+R. leguminosa um b . iceae (USDA 2478) and ISO44 signi ican ly supp essed O. c ena a haus o ia ac o as compa ed o he co esponding con ol. In he in i o expe imen , i espec i e o aba bean cul i a s and bac e ial inocula ion, O. c ena a eme gence p og essi ely inc eased wi h seed bank size. Faba been inocula ed wi h bac e ial combina ions o BMP+TAL1399 and BMP+USDA2478 signi ican ly educed O. c ena a eme gence and d y weigh . In gene al, Basabee cul i a in a iably displayed be e g ow h han Selaim. Faba bean heigh and d y weigh in a iably dec eased wi h inc easing seed bank size. The inc ease in aba bean g ow h pa ame e s is consis en wi h he obse ed delay and dec ease in O. c ena a eme gence in esponse o ea men s. CONCLUSION: Imp o ing soil e ili y by using bene icial mic oo ganisms appea ed o dec ease O. c ena a in es a ion and i s supp essi eness e ec s on hos g ow h. Use o po en ial bac e ial s ains and isola es could be inco po a ed in o exis ing O obanche c ena a managemen p ac ices. KEY WORDS B oom ape, Faba bean cul i a s, Soil bo ne bac e ia, Supp ession. INTRODUCTION Soil mic oo ganisms in e e ing wi h ea ly de elopmen al s ages o pa asi ic weeds we e hough o as possible al e na i es and/o iable supplemen s o o he con ol me hods [1].The symbio ic ela ionship o med be ween legumes and hizobia plays an in eg al ole in ag icul u e as bac e ia ix a mosphe ic ni ogen (N2). Rhizobia symbiosis wi h legumes p oduces 50% o 175 million ons o o al biological N2 ixa ion annually wo ldwide [2]. The e o e, inocula ion o legumes wi h e icien hizobia is one o he mos impo an and 31 ag onomically eco- iendly p ac ices used o imp o emen o N ixa ion [3]. Many mic oo ganisms possess an enzyma ic sys em which enables hem o mine alize phospho us-con aining o ganic compounds [4] o sec e e acids o solubilize ino ganic o solubilized compounds [5]. O obanche spp., O obanchaceae, is a majo biological cons ain o leguminous c ops p oduc ion in sub-Saha an [6]. A la ge pa o he damage is al eady done unde g ound e en be o e he pa asi e eme ges om he soil he eby making weeding ine ec i e op ion in i s con ol. Resea ch indings ha e asce ained he link be ween poo soil e ili y, s igolac ones p oduc ion and myco hizal in ec ion [7]. Nu ien de iciency is conduci e o s igolac ones biosyn hesis, educ ion o shoo b anching, maximiza ion o he symbio ic in e ac ions wi h a buscula myco hizal ungi(AMF) and acili a ion o nu ien s up ake. S igolac ones ha e been shown o unc ion as endogenous phy oho mones ha cu ail shoo b anching [8]. The p esen in es iga ions we e unde aken o e alua e he e ec s o bac e ial s ains and isola es cul u es and il a es, on ea ly de elopmen al s ages o he pa asi e and o explo e hei po en ial o deploymen as a componen o an in eg a ed managemen s a egy. MATERIALS AND METHODS Labo a o y expe imen s: A se ies o labo a o y expe imen s we e unde aken o in es iga e he e ec s o bac e ial s ains and isola es cul u es and il a es on ge mina ion and haus o ium ini ia ion o O. c ena a. T ea men s we e a anged in a Comple e Randomized Design (CRD) wi h 4 eplica es. All expe imen s we e epea ed h ee imes. Bac e ial isola es and s ains inoculums: Soil samples we e collec ed be ween No embe and Decembe , 2012 om Hudeiba Resea ch S a ion, Ag icul u al Resea ch Co po a ion (ARC), Sudan. The samples we e collec ed om he op 10 cm soil laye in aba bean ields and incuba ed on o a y shake (100 pm) a 28oC o 48 h o ob ain mic obial cul u es. Fo educing mic obial popula ion, one g am soil was dissol ed in 9 ml s e ile dis illed wa e . Se ial dilu ion was ca ied. Nu ien aga medium con aining an i ungal b illian g een was used o bac e ial g ow h. The medium was s e ilized by au ocla ing a 120ºC o 15min. The sp ead pla e me hod was used o isola ion o bac e ia. One ml o 10-5- 10-7 dilu ions we e pou ed and sp ead o e he media pla es using s e ile glass od. The dishes we e incuba ed a 28ºC o 48h.Mic obial colonies we e pu i ied by sub-cul u ing and we e mo phologically cha ac e ized. Fo y eigh bac e ial isola es we e ob ained om he soil samples. The isola ed bac e ia we e p ese ed in nu ien aga slan s and kep in a idge a 4ºC o u he s udies. E e y 3 mon hs in e al, bac e ial isola es we e e eshed. In addi ion, i e bac e ial s ains, Rhizobium leguminosa um b . iceae s ains (TAL1399, USDA 2478 and ENRRI9), Bacillus mega he ium a . Phospha icum (BMP), B. ci culans, Azomonas spp. and Fla obac e ium spp. we e ob ained om he Bio e iliza ion Depa men , En i onmen , Na u al Resou ces and Dese i ica ion Resea ch Ins i u e (ENDRI), he Na ional Cen e o Resea ch (NCR), Kha oum, Sudan. Plan ma e ials: O. c ena a seeds we e collec ed om in es ed aba bean ield in 2006 a Shendi Resea ch S a ion Fa m, Ag icul u e Resea ch Co po a ion (ARC) du ing he win e season. Seeds we e su ace disin ec ed by imme sion in 70% e hanol o 2-3 min., ollowed by washing h ee imes wi h s e ilized dis illed wa e . Then seeds we e imme sed wi h swi ling in o a 1% sodium hypochlo i e ob ained by app op ia e dilu ion o comme cial sodium hypochlo i e (Bleach) o 2-3 min. The sodium hypochlo i e was d ained o and he seeds we e washed unde suc ion sys em, wi h s e ilized dis illed wa e , se e al imes, un il he yellow colo disappea ed. The seeds, plo ed d y on Wha man No. 1 il e pape s, we e ai -d ied unde a lamina low cabine and subsequen ly s o ed a ambien empe a u e ill used. Chemicals p epa a ion: The s igolac one analogue GR24 was p o ided by p o esso Zwanenbe g, Uni e si y o Nimijhen, he Ne he lands. A s ock (10 ppm) o GR24 was p epa ed by dissol ing 1 mg in 1 ml ace one and comple ed o olume (100 ml) wi h s e ile dis illed wa e . The solu ion was kep e ige a ed a 4ºC o u he use. The haus o ium induce 2, 6 dime hoxybenzequinone syn he ic s imulan s (DMBQ) was p o ided by P o . Sugimo o, om Kobe Uni e si y, Japan. A s ock solu ion (100 ml) was p epa ed by dissol ing 1.68 gm in 1 ml ace one and comple ing o olume (100 ml) wi h s e ile dis illed wa e . The solu ion was kep e ige a ed a 4ºC o u he use. E ec s o bac e ial s ains and isola es cul u es on O. c ena a ge mina ion: Glass ibe il e pape s (GFFP) discs (8mm diame e ) we e cu , we ed ho oughly wi h wa e and placed in an o en (100 ºC o 1h.) o be s e ilized jus be o e use [9]. Fo p e-condi ioning, s e ilized discs, placed in 9 cm Pe i dishes lined wi h glass ibe il e pape s, we e mois ened wi h 5 ml dis illed wa e , b o h media yeas 32 ex ac manni ol, (mea pep one and nu ien b o h) o he espec i e bac e ial cul u e (TAL1399, USDA 2478, ENRRI9, BMP, B. ci culans, Azomonas spp., Fla obac e ium spp., ISO30, ISO33, ISO43 and ISO44). Abou 25-35, su ace disin ec ed O. c ena a seeds we e sp inkled on each o he glass ibe discs. The Pe i-dishes, sealed wi h pa a ilm and w apped in black poly hene, we e incuba ed in he da k a 18C o 11 days. Then each disc was subsequen ly ea ed wi h he syn he ic ge mina ion s imulan GR24 (25μl /disc) a 5 and 10ppm, we e e-incuba ed, and examined o ge mina ion a e 168h using s e eomic oscope. E ec s o bac e ial s ains il a e on O. c ena a ge mina ion: Two bac e ial s ains (TAL1399 and BMP) and hei combina ion we e sub-cul u ed in yeas ex ac Manni ol b o h medium and incuba ed o 24h a 30ºC. Bac e ial suspension was cen i uged a 6000 pm o 15 minu es and he supe na an was collec ed, il e ed and dilu ed wi h s e ile dis illed wa e o he desi ed concen a ions (25, 50, 75 and 100%). O. c ena a seeds we e mois ened wi h 5-ml o ei he s e ilized dis illed wa e o undilu ed o dilu ed bac e ial il a es and subsequen ly incuba ed and ea ed wi h GR24 as desc ibed abo e. E ec s o bac e ial s ains and isola es cul u es on O. c ena a haus o ial ini ia ion: Seeds condi ioned in selec ed bac e ial s ains and isola es cul u es (TAL1399, USDA 2478, ENRRI9, BMP, B. ci culans, Azomonas spp., Fla obac e ium spp., ISO30, ISO33, ISO43 and ISO44) we e induced o ge mina e wi h GR24. Discs con aining ge mina ed seeds (ge milings) we e blo ed d y on il e pape and placed op down on glass pape wi hou seeds. Each pai o discs was ea ed wi h 40μl o DMBQ. Then he dishes we e e-incuba ed. Seeds ans e ed o bac e ia ee media simila ly incuba ed and ea ed wi h GR24 we e included as con ol o compa ison. The Pe i dishes we e sealed wi h Pa a ilm, w apped in aluminum oil and incuba ed in he da k o 5 days. A seed was conside ed o ha e a haus o ium when he adical ips we e swollen and o med hai . E ec s o bac e ial s ains il a e on O. c ena a haus o ial ini ia ion: O. c ena a seeds we e ea ed wi h he selec ed bac e ia (TAL1399 o /and BMP) il a es a 25, 50, 75 and 100%.Then each disc was ea ed wi h GR24 o induce ge mina ion. Then ge milings we e ea ed wi h 40 μl o DMBQ a 10 and 20μM and subsequen ly incuba ed and examined o haus o ial ini ia ion as desc ibed abo e. Po expe imen : The expe imen was conduc ed in he g eenhouse o he Facul y o Ag icul u e, Omdu man Islamic Uni e si y, du ing Decembe , 2015 - Ma ch, 2016. The expe imen was conduc ed o s udy he e ec s o a ious combina ions comp ising bac e ial s ains and isola es, O. c ena a seed bank and esis an a ie ies on O. c ena a incidence and aba bean pe o mance. Plas ic po s (19 cm. diame e ), wi h d ainage holes a he bo om, we e illed wi h soil mix u e (9Kg/po ) o i e sil and sand (1:1 / ). A i icial in es a ion o soil was accomplished by mixing O. c ena a seeds (2g) wi h 1kg soil ollowed by subsequen dilu ions wi h O. c ena a ee soil o gi e he equi ed in es a ion le el (4 and 8 mg/ po ), O. c ena a in es ed and unin es ed con ols we e included o compa ison. Two bac e ial combina ions BMP+ TAL1399 and BMP+USDA2478 and wo aba been cul i a s (Selaim and Basabee ) we e used. Faba bean seeds (5/po ) we e sown a 2 cm soil dep h. The po s we e subsequen ly i iga ed e e y 2 days. Faba bean seedlings we e hinned o 2 plan s pe po a e 2 weeks o sowing. T ea men s we e a anged in a Randomized Comple e Block Design (RCBD) wi h ou eplica es. Da a collec ed o O. c ena a eme gences we e measu ed a 4, 6, 8, 9, 10, 11 and 12 weeks a e sowing (WAS). Da a collec ed o aba bean g ow h pa ame e s we e plan heigh and d y weigh . S a is ical analysis: P io o analyses da a on pe cen age (ge mina ion o haus o ium) we e a csine ans o med, da a on O. c ena a eme gence we e squa e oo ans o med o ul ill ANOVA equi emen s. The analyses we e pe o med ac oss expe imen s using SAS S a is ical package. Means sepa a ions we e made by he LSD a 5% [10]. Resul s: Labo a o y expe imen s: E ec o bac e ial s ains and isola es cul u e on O. c ena a ge mina ion: O. c ena a seeds ea ed wi h wa e displayed negligible ge mina ion (da a no shown). GR24 applied o O. c ena a seeds condi ioned in wa e induced he highes ge mina ion (58 - 69%) (Table1). Resul s e ealed ha condi ioning in he g ow h medium had no ad e se e ec on ge mina ion in esponse o GR24 a he lowe concen a ion. O. c ena a seeds, p e iously condi ioned in p esence o bac e ial s ains, we e compa able o ha o he co esponding nu ien b o h medium, i espec i e o ge mina ion s imulan . In among all bac e ia, B. ci culans s ain had he inhibi o ies e ec as compa ed o o he mic obes. The combina ion o BMP + TAL1399 consis en ly showed signi ican ly (P≤0.05) lowe ge mina ion han he co esponding nu ien b o h 33 con ol. The combina ion be ween B. ci culans + ENRRI 9 induced O. c ena a ge mina ion especially a he lowes concen a ion o GR24. Table 1: E ec s o bac e ial s ains and hei combina ions on O. c ena a ge mina ion in esponse o GR24 (du ing condi ioning) (Ba ch 1) T ea men s Ge mina ion (%) Means GR24 conc. (ppm) 5 10 Wa e 58.16* (72.00)** 69.97 (85.00) 63.97 Media (YEMB)# 51.12 (60.56) 54.16 (65.36) 52.60 ENRRI9 46.88 53.20) 44.22 (48.66) 45.55 TALL1399 38.98 (40.32) 46.17 (52.02) 42.57 USDA2478 42.62 (45.85) 42.53 (45.71) 42.57 Bacillus ci culans 37.90 (37.96) 45.96 (51.57) 41.93 BMP+ ENRRI9 48.33 (55.72) 47.74 (54.25) 48.03 BMP+TAL1399 33.73 (31.27) 35.78 (34.71) 34.75 BMP+USDA2478 34.66 (32.84) 37.43 (37.05) 36.04 B. ci culans + ENRRI9 67.17 (84.76) 57.44 (70.72) 62.30 B. ci culans + TAL1399 60.57 (75.36) 71.37 (85.47) 48.28 B. ci culans + USDA2478 42.21 (45.18) 45.69 (51.14) 43.95 LSD (Bac e ia) 8.28 LSD ( In e ac ion) 11.71 *Da a ou o b acke s a e a csine ans o med o analysis **Da a be ween b acke s a e o iginal da a # YEMB: Yeas Ex ac Manni ol B o h In he second ba ch, Seeds condi ioned in dis illed wa e and ea ed wi h GR24 a 5 and 10 ppm displayed 58.2 and 62.8% ge mina ion (Table 2). Resul s showed ha condi ioning in medium had no ad e se e ec on seed ge mina ion in esponse o GR24 a bo h concen a ions. Seeds condi ioned in BMP and simila ly ea ed wi h GR24 displayed low and di e en ial ge mina ion. The bac e ial combina ions o BMP plus Fla obac e ium spp. and BMP plus Azomonas spp. had no ad e se e ec s on O. c ena a ge mina ion in esponse o GR24. Table 2: E ec s o bac e ial s ains and hei combina ions on O. c ena a ge mina ion in esponse o GR24 (du ing condi ioning) (Ba ch 2) T ea men s Ge mina ion (%) Means GR24 conc.(ppm) 5 10 Wa e 58.22* (71.79)** 62.83 (78.76) 60.52 Media (MPB)# 52.99 (63.45) 59.47 (73.75) 56.23 BMP 40.05 (41.66) 42.57 (45.83) 41.31 Fla obac e ium spp. 48.84 (56.41) 51.07 (60.27) 49.96 Azomonas spp. 48.49 (55.93) 53.00 (57.04) 50.74 BMP + Fla obac e ium spp. 42.31 (45.47) 51.63 (61.25) 46.97 BMP + Azomonas spp. 51.83 (61.53) 56.43 (68.58) 54.13 LSD (Bac e ia) 10.06 LSD ( In e ac ion) 14.23 *Da a ou o b acke s a e a csine ans o med o analysis **Da a be ween b acke s a e o iginal da a # MPB: Mea Pep one B o h Resul s o ba ch 3, showed ha ge mina ion a e o seeds ea ed wi h GR24 was 56%. O. c ena a seeds, p e iously condi ioned in p esence o bac e ial isola es (ISO30, ISO44, ISO43 and ISO33), showed a iable esponse o GR24 (Table 3). In among all bac e ial isola es, ISO43 and ISO44 induced only 33.9 and 33.4% ge mina ion, espec i ely. Table 3: E ec o bac e ial isola es on O. c ena a ge mina ion in esponse o GR24 (du ing condi ioning) (Ba ch 3) T ea men s Ge mina ion (%) Means GR24 conc.(ppm) 5 10 Wa e 53.65* (57.26)** 58.84 (60.18) 56.24 Media (NB)# 52.24 (66.73) 53.74 (54.40) 52.99 ISO30## 38.07 (38.13) 46.64 (52.84) 42.35 ISO33 36.60 (35.71) 38.74 (39.55) 37.67 ISO43 34.19 (31.65) 33.65 (32.33) 33.92 ISO44 32.85 (30.10) 34.09 (30.45) 33.47 LSD (Bac e ia) 6.15 LSD ( In e ac ion) 8.69 *Da a ou o b acke s a e a csine ans o med o analysis **Da a be ween b acke s a e o iginal da a #NB: Nu ien B o h ##ISO: Bac e ial isola e 34 E ec s o bac e ial il a es on O. c ena a ge mina ion (du ing condi ioning): The p esen in es iga ion was se o s udy he e ec s o bac e ial il a es on O. c ena a ge mina ion (Table 4). Resul s displayed ha seeds condi ioned in he medium had no ad e se e ec on O. c ena a ge mina ion in esponse o GR24. The il a e o TAL1399 i espec i e o concen a ions, signi ican ly (P≤0.5) inhibi ed O. c ena a ge mina ion o 31.17-34.91% and 37.70-39.20% in esponse o GR24 a 5 and 10 ppm, espec i ely. The il a es o he combina ion o TAL1399 plus BMP educed O. c ena a ge mina ion by 5.05 -35.44% and 39.65-45.31% in esponse o GR24 a 5 and 10 ppm, espec i ely, compa ed o he con ol. While he BMP il a e a concen a ion o 75 and 100% signi ican ly (P≤0.5) inhibi ed O. c ena a ge mina ion by 44.3 and 37.7%, espec i ely. Table 4: E ec o bac e ial s ains il a es on O. c ena a ge mina ion in esponse o GR24 (du ing condi ioning) Fil a ion Fil a e concen a ion Ge mina ion (%) Means GR24 conc. (ppm) 5 10 Wa e 66.45* (83.64)** 67.40 (84.68) 66.92 Media (YEMB)# 67.57 (84.97) 69.23 (86.63) 68.40 BMP 100 36.38 (35.50) 39.10 (39.97) 37.74 75 44.80 (49.63) 43.84 (48.00) 44.32 50 50.76 (59.80) 53.33 (64.05) 52.04 25 55.78 (68.32) 57.25 (69.93) 56.51 TAL1399 100 40.40 (42.03) 41.99 (44.77) 41.19 75 43.25 (46.99) 47.06 (53.48) 45.15 50 46.19 (52.06) 47.55 (54.42) 46.87 25 54.70 (66.38) 56.19 (69.02) 55.44 BMP + TAL1399 100 36.34 (35.18) 40.67 (42.49) 38.50 75 42.90 (46.45) 43.77 (47.99) 43.33 50 47.54 (54.32) 50.48 (59.49) 49.01 25 49.11 (56.87) 52.77 (63.09) 50.94 LSD (Bac e ial il a e) 2.59 LSD (Fil a e concen a ion) 2.59 LSD ( In e ac ion) 8.18 *Da a ou o b acke s a e a csine ans o med o analysis **Da a be ween b acke s a e o iginal da a # YEMB: Yeas Ex ac Manni ol B o h E ec o bac e ial s ains and isola es on O. c ena a haus o ium ini ia ion: Resul s e ealed ha O. c ena a ge milings esul ing om seeds condi ioned in wa e o nu ien b o h medium showed simila esponse o DMBQ (Table 5). In among he bac e ial s ains, he inhibi o y e ec was highes in B. ci culans as compa ed o o he ea men s. Howe e , condi ioning in USDA2478 esul ed in a no iceable inc ease in haus o ium ini ia ion, albei no signi ican compa ed o he b o h medium. Condi ioning in BMP + USDA2478 educed haus o ium ini ia ion and he obse ed educ ion inc eased wi h DMBQ concen a ion. Table 5: E ec s o bac e ial s ains and hei combina ions on O. c ena a haus o ium in esponse o DMBQ (du ing condi ioning) (Ba ch 1) T ea men s Haus o ium (%) Means DMBQ conc. (µM) 10 20 Wa e 44.04* (48.33)** 45.98 (51.70) 45.01 Media (YEMB)# 41.83 (44.48) 45.25 (50.43) 43.54 ENRRI9 41.92 (44.72) 33.04 (30.23) 37.48 TALL1399 49.53 (57.51) 41.34 (43.69) 45.43 USDA2478 47.36 (54.18) 48.97 (56.76) 48.16 Bacillus ci culans 27.11 (21.33) 39.86 (41.17) 33.48 BMP+ ENRRI9 43.64 (47.76) 35.25 (33.53) 39.44 BMP+TAL1399 33.04 (29.85) 33.23 (30.18) 33.13 BMP+USDA2478 29.32 (42.45) 31.30 (27.77) 30.31 B. ci culans + ENRRI9 38.59 (39.07) 45.68 (51.56) 42.13 B. ci culans + TAL1399 43.04 (46.21) 43.22 (46.96) 43.13 B. ci culans + USDA2478 39.84 (41.59) 37.83 (38.12) 38.83 LSD (Bac e ia) 8.47 LSD ( In e ac ion) 11.98 *Da a ou o b acke s a e a csine ans o med o analysis **Da a be ween b acke s a e o iginal da a #YEMB: Yeas Ex ac Manni ol B o h O. c ena a ge mlings a ising om seeds condi ioned in dis illed wa e and b o h medium and subsequen ea ed wi h GR24 displayed 51.9% and 43.6%, haus o ium ini ia ion in esponse o DMBQ espec i ely (Table 6). Condi ioning in BMP cul u e signi ican ly (P≤0.05) educed haus o ium ini ia ion and he obse ed educ ion 35 inc eased wi h concen a ion. The combina ions o he bac e ial s ains signi ican ly (P≤0.05) inhibi ed haus o ium induc ion compa ed o he co esponding wa e con ol. Table 6: E ec o bac e ial s ains and hei combina ions on O. c ena a haus o iumin esponse o DMBQ (Ba ch 2) T ea men s Haus o ium (%) Means DMBQ conc.(µM) 10 20 Wa e 48.90* (56.65)** 54.92 (66.83) 51.91 Media (MPB)# 42.65 (46.18) 44.60 (49.31) 43.62 BMP 37.64 (37.49) 32.59 (30.50) 35.11 Fla obac e ium spp. 50.17 (58.54) 42.14 (45.07) 46.15 Azomonas spp. 46.09 (51.80) 41.29 (43.58) 43.69 BMP + Fla obac e ium spp. 34.41 (32.46) 34.22 (31.65) 34.31 BMP + Azomonas spp. 29.95 (25.29) 34.67 (32.50) 32.31 LSD (Bac e ia) 7.00 LSD ( In e ac ion) 9.89 *Da a ou o b acke s a e a csine ans o med o analysis **Da a be ween b acke s a e o iginal da a #MPB: Mea Pep one B o h DMBQ a 10 and 20 µM applied o O. c ena a ge milings esul ing om seeds p e iously condi ioned in wa e and ea ed wi h GR24 induced 55.7% and 59.3% haus o ia, espec i ely (Table 7). Seed condi ioned in nu ien b o h medium, s imula ed o ge mina e wi h GR24 and simila ly ea ed wi h DMBQ displayed 52.4% and 53.6% haus o ia, espec i ely. In among he bac e ial isola es, ISO44 signi ican ly (P≤0.05) educed haus o ial ini ia ion, i espec i e o haus o ium inducing ac o . Table 7: E ec s o bac e ial isola es on O. c ena a haus o iumin esponse o DMBQ (Ba ch 3) T ea men s Haus o ium (%) Means DMBQ conc.(µM) 10 20 Wa e 55.70* (68.00)** 59.25 (73.26) 57.47 Media (NB)# 52.44 (62.77) 53.62 (64.58) 53.03 ISO 30## 34.36 (32.22) 38.10 (38.33) 36.23 ISO 33 34.67 (32.41) 39.39 (40.34) 37.03 ISO 43 33.16 (30.41) 34.05 (31.63) 33.60 ISO 44 28.90 (23.71) 30.31 (25.56) 29.60 LSD (Bac e ia) 5.87 LSD ( In e ac ion) 8.31 *Da a ou o b acke s a e a csine ans o med o analysis **Da a be ween b acke s a e o iginal da a #NB: Nu ien B o h ##ISO: Bac e ial isola e E ec s o bac e ial il a es on O. c ena a haus o ium ini ia ion: Fil a es om he combina ion o TAL1399 + BMP signi ican ly (P≤0.5) inhibi ed he haus o ium ini ia ion in esponse o DMBQ a 10 and 20 µM, i educed haus o ium by 44.80% and 41.68%, ollowed by BMP which educed haus o ium ini ia ion by 58.61%and 59.03% espec i ely, i espec i e o he il a e concen a ions, compa ed o con ol (Table 8). Table 8: E ec o bac e ial s ains il a es on O. c ena a haus o ium in esponse o DMBQ Fil a ion Fil a e concen a ion Haus o ium (%) Means DMBQ conc.(µM) 10 20 Wa e 68.11* (86.03)** 68.82 (86.80) 68.46 Media (YEMB)# 64.82 (81.68) 70.71 (88.75) 67.76 BMP 100 34.06 (31.45) 35.56 (33.81) 34.81 75 36.96 (36.81) 42.82 (46.21) 39.89 50 43.61 (47.57) 44.35 (48.87) 43.98 25 48.31 (55.76) 49.69 (58.01) 49.00 TAL1399 100 37.70 (37.41) 39.11 (39.79) 38.40 75 39.73 (40.87) 41.22 (43.41) 40.47 50 41.65 (44.17) 44.70 (49.47) 41.65 25 45.91 (51.58) 48.20 (55.56) 47.05 BMP + TAL1399 100 28.19 (22.37) 28.19 (22.58) 28.19 75 37.59 (37.31) 40.13 (41.59) 38.86 50 41.97 (44.73) 44.06 (48.36) 43.01 25 44.75 (49.56) 46.14 (51.98) 45.44 LSD (Bac e ial il a ion) 1.40 LSD (Fil a e concen a ion) 1.40 LSD ( In e ac ion) 4.44 *Da a ou o b acke s a e a csine ans o med o analysis. **Da a be ween b acke s a e o iginal da a. # YEMB: Yeas Ex ac Manni ol B o h. 36 G een house expe imen : E ec s o bac e ial s ains, cul i a s and O. c ena a seeds le el on O. c ena a incidence in aba bean: A 8 WAS, a ew eme gences o O. c ena a was obse ed, i espec i e o seed bank size and aba bean cul i a s (Table 9). A 9 WAS, i espec i e o he seed bank size, esul s showed conside able inc ease in compa ison o ha eco ded a 8 WAS. A 10 WAS, O. c ena a eme gence on aba bean cul i a s inocula ed wi h BMP+TAL1399 and BMP+USDA2478 sus ained he lowes eme gence, i espec i e o seeds bank size as compa ed o he con ol. A 11 WAS, a u he inc ease in O. c ena a seed bank o 8 mg pe po inc eased he pa asi e eme gence. Table 9: E ec s o bac e ial s ains and aba bean cul i a s on O. c ena a incidence T ea men s O. c ena a coun Faba bean cul i a s Bac e ia O. c ena a seeds conc. (mg/ po ) Weeks a e sowing Means 8 9 10 11 12 Selaim Con ol 4 0.71*(0.00)** 1.56 (2.00) 1.93 (3.25) 2.28 (4.75) 2.81 (7.50) 1.86 8 0.97 (0.50) 2.00 (3.50) 2.11 (4.00) 2.34 (5.00) 2.59 (6.25) 2.00 BMP+TAL 1399 4 0.71 (0.00) 1.73 (2.50) 1.73 (2.50) 1.56 (2.00) 2.06 (3.75) 1.56 8 0.93 (0.50) 1.82 (3.00) 2.10 (4.50) 1.86 (3.75) 1.86 (3.75) 1.71 BMP+USD A2478 4 0.71 (0.00) 1.68 (2.50) 1.68 (2.50) 1.59 (2.25) 1.59 (2.25) 1.45 8 0.84 (0.25) 1.19 (1.25) 1.58 (2.75) 1.84 (3.25) 1.84 (3.25) 1.46 Basabee Con ol 4 0.93 (0.50) 1.98 (3.50) 2.10 (4.00) 2.27 (4.75) 2.81 (7.50) 2.02 8 1.06 (1.00) 2.11 (4.00) 2.27 (4.75) 2.48 (5.75) 2.66 (6.75) 2.12 BMP+TAL 1399 4 0.84 (0.25) 1.64 (2.50) 1.57 (2.25) 1.57 (2.25) 1.57 (2.25) 1.44 8 0.71 (0.00) 1.27 (1.20) 1.50 (2.00) 1.64 (2.25) 1.64 (2.25) 1.35 BMP+USD A2478 4 0.71 (0.00) 1.86 (3.00) 1.86 (3.00) 1.89 (3.25) 2.39 (5.25) 1.74 8 0.71 (0.00) 1.54 (2.25) 1.70 (3.00) 1.92 (3.50) 1.92 (3.50) 1.56 Means 0.82 1.70 1.84 1.94 2.14 LSD 0.50 0.73 0.95 0.88 0.88 * indica es squa e oo ans o med da a (√x+0.5 x: a iable) **Da a be ween b acke s a e o iginal da a Faba bean cul i a s inocula ed wi h bo h bac e ial combina ions dec eased O. c ena a eme gence albei no signi ican ly in p esence o he wo O. c ena a seed bank size, compa ed o he con ol. A 12 WAS, Basabee cul i a inocula ed wi h bo h bac e ial combina ions signi ican ly (P≤0.5) educed O. c ena a eme gence a he wo seed bank size. While Seliam cul i a ea ed wi h BMP+TAL1399 sus ained he lowes O. c ena a eme gence a he lowe seed bank size (4mg/po ) compa ed o he con ol. In gene al, Selaim cul i a sus ained he lowes O. c ena a in es a ion as compa ed o Basabee , i espec i e o O. c ena a seed bank and bac e ial inocula ion. Inocula ion wi h he bac e ial combina ions BMP+TAL1399 and BMP+USDA2478 educed O. c ena a eme gence, i espec i e o aba bean cul i a s and he pa asi e seed bank. E ec s o bac e ial s ains, cul i a s and O. c ena a seed bank on aba bean g ow h: In all ea men s, aba bean heigh p og essi ely dec eased wi h inc ease in O. c ena a seed bank size and inc eased wi h ime (Table 10). Basabee cul i a , i espec i e o bac e ial inocula ion o O. c ena a seed bank size displayed be e g ow h han Selaim cul i a . Mo eo e , aba bean inocula ed wi h bac e ia BMP+TAL1399 and BMP+USDA2478, i espec i e o cul i a s and seed bank size, displayed be e g ow h han un-inocula ed con ol. A 4 WAS, O. c ena a seed bank a 4mg/po in lic ed insigni ican dec ease in Basabee plan heigh . Inc easing O. c ena a seed bank size o 8 mg pe po signi ican ly (P≤0.05) educed aba bean heigh by 20.02 %. A 6 WAS, Selaim cul i a inocula ed wi h BMP plus TAL1399 in p esence o he O. c ena a seed bank a 8 mg seeds/po inc eased aba bean heigh albei no signi ican ly compa ed o he con ol. Howe e , Basabee cul i a ea ed wi h he same combina ion ga e he highes g ow h a lowe le el o O. c ena a seed bank. A 8 WAS, inocula ion wi h BMP+TAL1399 a he lowe O. c ena a in es a ion seed bank size (4mg/po ) signi ican ly (P≤0.5) inc eased Selaim plan heigh compa ed o he co esponding con ol. A 9 WAS Selaim cul i a inocula ed wi h he bac e ial combina ion o BMP+TAL1399 displayed he highes g ow h as compa ed o he con ol, i espec i e o O. c ena a seeds bank. A 10, 11 and 12 WAS aba bean heigh , i espec i e o cul i a s and bac e ial inocula ion, displayed a p og essi e dec ease wi h O. c ena a seed bank size. Inocula ion wi h BMP+USDA2478 signi ican ly (P≤0.5) inc eased Selaim cul i a plan heigh a he lowe seed bank size a 10 and 11 WAS compa ed o he co esponding con ol. 37 Table 10: E ec s o bac e ial s ains and O. c ena a on aba bean g ow h T ea men s Plan heigh (cm) Means Faba bean cul i a s Bac e ia O. c ena a seeds le el (mg/ po ) Weeks A e Sowing (WAS) 4 6 8 9 10 11 12 Selaim Con ol 0 22.08 30.50 34.45 38.00 45.66 46.12 47.33 37.73 4 23.66 33.41 35.85 38.33 40.08 38.58 40.46 35.77 8 22.49 32.62 38.87 40.99 41.66 42.25 37.75 36.66 BMP+ TAL1399 4 19.75 32.45 37.75 39.99 40.16 42.66 41.12 36.27 8 24.41 35.29 42.58 44.41 43.08 45.50 39.58 39.26 BMP+ USDA2478 4 22.16 33.58 40.16 41.16 44.50 43.75 43.54 38.41 8 21.66 34.08 39.00 40.66 41.75 44.70 40.25 37.44 Basabee Con ol 0 24.58 34.91 42.50 44.58 52.99 53.66 56.25 44.21 4 21.54 33.04 41.83 43.20 44.08 44.37 40.29 38.34 8 19.66 32.83 41.58 43.16 43.66 44.50 40.75 38.02 BMP+ TAL1399 4 26.12 36.50 43.08 43.75 45.08 43.83 40.04 39.77 8 23.50 34.50 39.25 40.00 42.00 41.75 38.00 37.00 BMP+ USDA2478 4 20.62 32.95 39.25 39.95 40.87 41.08 41.21 36.56 8 20.41 32.20 39.62 40.87 41.79 42.45 38.33 36.52 Means 22.33 33.49 39.70 41.36 43.38 43.94 41.78 LSD 3.55 3.56 4.15 3.99 4.10 3.84 4.01 E ec s o bac e ial s ains, cul i a s and O. c ena a seeds bank on aba bean d y weigh : Faba bean d y weigh i espec i e o bac e ial combina ions and cul i a s, dec eased wi h inc easing O. c ena a seeds bank size (Table11). O. c ena a seed bank size a 4 and 8 mg/po educed aba bean biomass by 8.07 and 16.95% espec i ely as compa ed o unin es ed con ol. Resul s showed ha inocula ion o Selaim cul i a wi h he bac e ial combina ions did no a ec he plan d y weigh . Inocula ion wi h BMP+USDA2478 signi ican ly (P≤0.5) inc eased Basabee cul i a oo d y weigh a he lowe seed bank size. While inocula ion wi h BMP+TAL1399 signi ican ly (P≤0.5) inc eased Basabee shoo d y weigh , in p esence o O. c ena a seed bank a 4 mg/po compa ed o in es ed con ol. Table 11: E ec s o bac e ial s ains and O. c ena a on aba bean d y weigh T ea men s D y weigh (g) Faba bean cul i a s Bac e ia O. c ena a seeds conc. (mg/ po ) Roo Shoo To al biomass Selaim Con ol 0 2.95 15.40 9.18 4 2.82 14.05 8.44 8 1.87 13.37 7.62 BMP+TAL1399 4 2.62 9.57 6.10 8 2.00 11.87 6.94 BMP+USDA2478 4 1.85 7.47 4.66 8 1.65 7.70 4.68 Basabee Con ol 0 3.95 17.07 10.51 4 1.32 8.10 4.71 8 1.80 14.52 8.16 BMP+TAL1399 4 1.12 12.02 6.57 8 2.50 10.95 6.73 BMP+USDA2478 4 2.55 11.27 6.91 8 1.07 6.97 4.02 Means 2.26 12.11 LSD 0.91 3.64 Discussion: O obanche c ena a Fo sk is a majo cons ain o aba bean (Vicia abaL.) p oduc ion in oduced ecen ly in o he Sudan and widely dis ibu ed and became a na ional p oblem. The s udy showed ha in labo a o y and po expe imen s, bac e ial s ains and isola es ha e he po en ial o educe O. c ena a in es a ion and mi iga e a leas in pa i s nega i e e ec s on g ow h o aba bean [11]. The p esen s udy e ealed ha ge mina ion o O. c ena a inc eased wi h inc easing GR24 concen a ion. Ge mina ion o O. c ena a dec eased signi ican ly a e inocula ion wi h bac e ial cul u es o il a es o ISO43 and ISO44 isola es and B. ci culans, BMP, BMP+TAL1399 s ains as compa ed o o he mic obes and con ols. Hassan and Abakee [9] in simila s udy epo ed ha he combina ions o BMP plus TAL1399 and BMP plus USDA2478 inhibi ed O obanche ge mina ion. Ga a e al. [12] epo ed ha isola e ISO20 (Gluconace oac e spp.) had he po en ial o inhibi S iga ge mina ion and adical elonga ion. The decline in ge mina ion o O. c ena a wi h he bac e ia could be due o a possible phy o oxic e ec on he emb yo hus leading o educ ion o i s ou wa ds h us on he su ounding issues. Wi h espec o he e ec s o bac e ial cul u es o il a es on haus o ium ini ia ion, esul s 38 displayed ha Bacillus ci culans, BMP, BMP+USDA2478 and isola e ISO 44 supp essed O. c ena a haus o ia ac o signi ican ly as compa ed o he co esponding con ol. Ba ghou hi and Salman [13] epo ed ha bac e ia may con ol weeds by in e up ing signals equi ed o adical elonga ion, haus o ium o ma ion, hizo opism o a achmen . In he g eenhouse expe imen , esul s displayed ha seed bank size in luenced pa asi ism and g ow h pa ame e s o bo h he hos and he pa asi e. In es a ion o O. c ena a o aba bean, in a iably, inc eased wi h inc easing size o he seed bank. In among he wo aba bean cul i a s es ed, basabee showed he highes O. c ena a eme gence as compa ed o Selaim. O. c ena a, i espec i e o he seed bank size and cul i a s educed plan heigh and d y ma e . These indings a e consis en wi h hose ob ained by Mesa-Ga cia and Ga cia- To es [14] who epo ed ha O. c ena a in ec ion esul ed in signi ican dec ease in aba bean. The p esen s udy showed ha bac e ia ha e he po en ial o educe O. c ena a pa asi ism and damage o aba bean g ow h. This esul is ongoing wi h Es a e al. [15] who epo ed ha inocula ion cowpea wi h Rhizobium leguminosa um delayed and ep essed S iga eme gence. Mo eo e , aba bean inocula ed wi h bac e ial BMP+TAL1399 and BMP+USDA2478, i espec i e o cul i a s and O. c ena a seed bank size, displayed be e g ow h han un-inocula ed c op. Basabee cul i a , i espec i e o bac e ial inocula ion o O. c ena a seed bank size displayed be e g ow h han Selaim cul i a . Ahonsi e al. [16] epo ed ha co-inocula ion o legumes wi h e hylene-p oducing pseudomonads and N2- ixing b ady hizobial s ains as supplemen s o legume o a ion, educed S iga in es a ion in maize The educ ions in shoo and oo biomass a e consis en wi h hose p e iously epo ed by F os e al. [17] and could be a ibu ed o a mul i ude o ac o s ela ed o d y ma e p oduc ion and pa i ioning including siphoning o nu ien s, wa e and pho osyn ha e by he pa asi e. I is known ha hizosphe e bac e ia a e capable o p oducing compounds, which i aken up by plan s, can s imula e de ense esponses agains dele e ious pa hogens [18]. Musyoka [19] showed ha inocula ing cowpeas wi h Rhizobium in soils supplied wi h P has he po en ial o inc ease soil a ailable N. Imp o ing soil e ili y as epo ed by Jain and Foy [20] appea s o dec ease O obanche in es a ion and i s enhancemen on hos g ow h. Conclusion: The esul s clea ly showed he ad e se e ec s o O. c ena a on i s hos and he need o an in eg a ed app oach o O. c ena a managemen . Howe e , hese esul s need o be e i ied in ield expe imen s and he cos e ec i eness o he ea men s needs o be conside ed. REFERENCES [1] Saue bo n, J., D. Mulle -S o e and J. He shenho n, 2007. The ole o biological con ol in managing pa asi ic weeds. CROP PROT., 26: 246-254. [2] Yada , J.and J.P. Ve ma, 2014. E ec o seed inocula ion wi h indigenous Rhizobium and plan g ow h p omo ing hizobac e ia on nu ien s up ake and yields o chickpea (Cice a ie inum L.). EUR. J. SOIL BIOL., 63: 70-77. [3] Den on, M.D., D.J. Pea ce and M.B. Peoples, 2013. Ni ogen con ibu ions om aba bean (Vicia aba L.) elian on soil hizobia o inocula ion. PLANT AND SOIL., 365: 363-374. [4] Kannaiyan, S., K. Kuma and K. Go inda ajan, 2004. Bio e ilize Technology o Rice Based C opping Sys em. Scien i ic pub., Jodhpu , India. [5] Elsheikh, E.A.E., 1993. Soil Mic obiology. Kha oum Uni e si y P ess. Kha oum, Sudan. pp 256. (in A abic). [6] Wes wood, J.H., J.I. Yode , M.P. Timko and C.W. Pamphilis, 2010. The e olu ion o pa asi ism in plan s. T ends in Plan Science, 15(4): 227-235. [7] Akiyama, K., K. Ma suzaki and H. Hayashi, 2005. Plan sesqui e penes induce hyphal b anching in a buscula myco hizal ungi. NATURE., 435: 824-827. [8] Gomez-Roldan, V., S. Fe mas, P.B. B ewe , V. Puech-Pagès, E.A. Dun and J.P. Pillo , 2008. S igolac one inhibi ion o shoo b anching. NATURE., 455:189-194. [9] Hassan, M.M.and R.A. Abakee , 2013. E ec s o a buscula myco hizal ungi (AMF) and bac e ial s ains on O obanche c ena a Fo sk.on aba bean. UNIVERSAL JOURNAL OF APPLIED SCIENCE., 1(1): 27- 32. [10] Gomez, K.A.and A.A. Gomez, 1984. S a is ical P ocedu es o Ag icul u al Resea ch. 2nd Edi ion, John Wiley and Sons, New Yo k. [11] Babike ,A.G.T., E.A. Ahmed, D.A. Dawoud and N.K. Abdella, 2007. O obanche species in Sudan: His o y, Dis ibu ion and managemen . SUDAN JOURNAL OF AGRICULTURAL RESEARCH, 10: 107-114. [12] Ga a , N.Y., M.M. Hassan, M.M. Ahmed, A.G. Osman, M.E. Abdelgani and A.E. Babike , 2016. In i o s udy o endophy ic bac e ia, ca bohyd a es and hei combina ion on ea ly de elopmen al s ages o S iga he mon hica (Del.) Ben h. ADVANCES IN ENVIRONMENTAL BIOLOGY., 10(6): 66-74.