Isolamento de fungos fitopatogênicos em sementes da árvore Caatinga

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1 Revist de l Fcultd de Agronomí, L Plt (2017) Vol 116 (2): Isolmento de fungos fitoptogênicos em sementes d árvore Cting Do Nscimento, Mri ds Grçs Rodrigues 1 ; Kilson Pinheiro Lopes; Márci Aprecid Cezr; Mgly Morgn Lopes d Cost; Tigo Augusto Lim Crdoso ; Mri Gilmr de Oliveir Sores Unidde Acdêmic de Ciêncis Agráris, CCTA/UFCG; 1 grc.gronomi@gmil.com Do Nscimento, Mri ds Grçs Rodrigues; Kilson Pinheiro Lopes; Márci Aprecid Cezr; Mgly Morgn Lopes d Cost; Tigo Augusto Lim Crdoso; Mri Gilmr de Oliveir Sores (2017) Isolmento de fungos fitoptogênicos em sementes d árvore Cting. Rev. Fc. Agron. Vol 116 (2): A qulidde snitári é um prâmetro que vli incidênci de ptógenos em sementes, principlmente em espécies de produção grícol, hvendo poucos estudos sore espécies florestis róres. O presente trlho tem como ojetivo vlir qulidde snitári em sementes de espécies róres d Cting. O experimento foi conduzido no Lortório de Fitoptologi do Centro de Ciêncis e Tecnologi Agrolimentr d UFCG, cmpus de Poml-PB. Form estudds sementes de qutro espécies róres: Myrcrodruon urundeuv (Allemão): roeir, Poincinell pyrmidlis (Tul.): ctingueir, Mimos tenuiflor (Willd.)Poiret: jurem-pret e Liidii ferre (Mrt) ex Tul.: pu-ferro, s mesms form dividids em dus sumostrs, sendo um sumetid à desinfestção superficil por imersão em hipoclorito de sódio (NClO) 2% por 5 minutos. A snidde ds sementes foi vlid pelo lotter test. Os gêneros fúngicos comumente detectdos ns sementes ds espécies estudds form: Aspergillus spp., Cldosporium spp., Fusrium spp., Penicillium spp. e Trichoderm spp.. A desinfestção com hipoclorito de sódio 2% reduziu incidênci de fungos ns sementes ds espécies estudds. O gênero Aspergillus presentou o mior número de colônis e diversidde de espécies fúngics ns sementes trtds e não trtds, estndo presente em tods s espécies róres. A menor incidênci de colônis fúngics ocorreu n espécie Mimos tenuiflor. Plvrs-chve: sementes florestis, snidde, desinfestção, micoflor, diversidde. Do Nscimento, Mri ds Grçs Rodrigues; Kilson Pinheiro Lopes; Márci Aprecid Cezr; Mgly Morgn Lopes d Cost; Tigo Augusto Lim Crdoso; Mri Gilmr de Oliveir Sores (2017) Isoltion of phytopthogenic fungi in seeds Cting tree. Rev. Fc. Agron. Vol 116 (2): The snitry qulity is prmeter tht evlutes the incidence of pthogens in seeds, especilly in species of griculturl production, so tht there re few studies on forest tree species. This study ims to evlute the snitry qulity nd the effect on seeds of four species from the Cting region. The experiment ws conducted t the Lortory of Phytopthology from the Centre for Agri-Food Science nd Technology t UFCG, Poml - PB. Seeds from four species were studied: Myrcrodruon urundeuv (Allemão): roeir, Poincinell pyrmidlis (Tul.) L.P. Queiroz: ctingueir, Mimos tenuiflor (Willd.) Poiret: jurem-pret nd Liidii ferre (Mrt ex Tul.) L.P. Queiroz: pu-ferro. The seed helth ws ssessed using the lotter test. The gener of fungi commonly found in the seeds of the studied species were Aspergillus spp., Cldosporium spp., Fusrium spp., Penicillium spp. nd Trichoderm spp. The disinfection with 2% sodium hypochlorite reduced the incidence of fungi on seeds of the studied species. The genus Aspergillus presented the highest numer of colonies nd diversity of fungl species in treted nd untreted seeds, eing present in ll four species. The lowest incidence of fungl colonies occurred in the species Mimos tenuiflor. Keywords: forest seeds, seed helth, disinfesttion, micoflor, diversity. Reciido: 25/09/2016 Aceptdo: 02/11/2017 Disponile on line: 01/04/2018 ISSN ISSN (on line) , Fcultd de Ciencis Agrris y Forestles, UNLP, Argentin 241

2 Do Nscimento et l (2017) INTRODUCTION The ssocition of pthogens to seed implies reduction of yield nd impirment of crop qulity. So, the ssessment of snitry qulity of seeds plys key role in the selection of resistnt cultivrs nd possile pre-tretments (Mrtins et l., 2009). In tropicl regions, the environmentl conditions of high humidity nd temperture re fvorle for the growth nd development of pthogens so tht the seeds of ntive species in these res re prone to e ttcked y them (Nscimento et l., 2006). In ddition to the reduction in germintion of seed lot, the presence of fungi my ring out prolems in interpreting the results of the germintion tests under lortory conditions (Sntos et l., 2011). Fungi re considered primry pthogen tht cuses dmge when ssocited with seeds. Dhingr et l. (2002) report Pestlotiopsis mculns s potentilly pthogenic to seeds nd seedlings of Andennther mcrocrp (mimos). In the forestry field, loss of ntive species seeds is common due to the ttck of sprophytic nd wooddecy fungi resulting from indequte storge conditions. Alfens nd Mfi (2007) ttriuted this to two fctors: (1) the fruits open in the field efore hrvesting, leving the seeds exposed to contmintion; nd (2) due to the height of trees, the fruits re collected on the ground, where fungi my colonize them nd rech the seeds. The pthogens trnsmitted or not y seeds cn lso ffect their force field, presenting more pronounced effect when they consist of orgnisms tht colonize the internl tissues of the seeds (Fgn et l., 2004). This type of informtion is essentil to projects focusing on the reforesttion with ntive tree species (Sntos et l., 2011). In ddition to the significnt losses t the field level, other prolems rise when infected seeds nd kernels re used in oth humn nd niml food, considering the fct tht fungi cn produce toxins nd there re high incidences of fungl contminnts oserved in mny types of seed storge. These re concerning dt since they directly ffect the physiologicl qulity of seeds (Silv et l., 2010). Looking for tree seeds for reforesttion with preservtionists purposes or not hs intensified in recent yers. Thus, the exchnge of seeds etween regions hs een expnded nd my constitute men of pthogens movement since this llows their dissemintion to the long distnces (Sntos et l., 2011). Currently, there re no products in Brzil to tret seeds of tree nd forest species for control pthogens, nor ny stisfctory result of reserch supports this method. So, the study of helth qulity of these species ims to contriute to knowledge nd technologies for the production system of ntive nd exotic tree seeds. Studying the ssocition of fungi with tree species my provide sis for epidemiologicl models, rnging from the storge of seed to seedling production (Sntos et l., 2011). Therefore, this study ims to ssess the helth nd the effect of surfce disinfection on the snitry qulity of seeds of tree species from the Cting region. Fungos fitoptogênicos em sementes d árvore Cting MATERIAL AND METHODS We crried out the experiment t the Lortory of Phytopthology, Centre for Agrifood Science nd Technology, Federl University of Cmpin Grnde, Poml PB. Seeds or propgules with full physiologicl mturity were hrvested from trees mtrices locted in the city of Aprecid, PB. After the crop, the seeds were extrcted from fruits, nd sujected to clening nd mnul selection to eliminte from those formed poorly or ttcked y insects, put to dry nd stored in glss pots with lortory temperture rnging from 18 to 25 C for yer. To ssess the tretment effect of the surfce disinfection on the incidence of fungi, seeds of four species Myrcrodruon urundeuv (Allemão): roeir, Poincinell pyrmidlis (Tul.): ctingueir, Mimos tenuiflor (Willd.) Poiret: jurem-pret nd Liidii ferre (Mrt) ex Tul.: pu-ferro were sorted into two susmples, one of them hving een sujected to disinfection y immersion in 2% sodium hypochlorite (NClO) remining in contct with the seed for 5 minutes. Seeds were pcked in gs of tulle, nd then the excess ws removed with their immersion in sterile distilled wter, wheres the remining susmple did not undergo ny tretment. For helth ssessment, we pplied the lotter test (Neegrd, 1979). Treted nd untreted seeds were plced in Petri dishes (150 mm of dimeter nd 20 mm height) contining three sheets of filter pper moistened with 15 ml of sterile distilled wter. The seeds were incuted t 23 ±2 C with 12 hours of photoperiod during ten dys (Oliveir, 2009). After the incution period, the seeds were individully exmined under stereoscopic microscope for the detection nd identifiction of ssocited fungi therewith (Brnett nd Hunter, 1998; Mp, 2009; Sntos et l., 2011). The results of the tles refer to the rw dt nd re lso expressed s percentges, the figures show the mens nd stndrd errors per repetition, tht is, the "worked" dt. When we were unle to identify the pthogens on seeds, they were isolted in the culture medium PDA + A (potto-dextrose-gr + ntiiotic) nd incuted t 28 C with 12 hours photoperiod for ten dys. The complete rndomized design ws used with two tretments (with nd without disinfection) nd ten repetitions of 20 seeds per plte, totlizing 200 seeds per tree species. We pplied the Kruskl-Wllis test compring two or more pired groups (nonprmetric ANOVA) to verify the occurrence of significnt differences mong species concerning the following vriles: numer of fungl colonies, numer of species nd fungl gener, nd numer of seeds germinted. The vriles tht showed significnt differences were sujected to the multiple comprison tests. The Wilcoxon comprison test, which verifies the mgnitude of the difference etween two tretments, ws pplied to confirm if the tretment with sodium hypochlorite ffected the vriles ove to show the efficiency of sodium hypochlorite for disinfesttion of tree seeds. A Correspondence Anlysis ws used to investigte the occurrence of ffinities etween the studied tree species concerning their susceptiilities to given set of fungl species. All nlyses were performed using R

3 Revist de l Fcultd de Agronomí, L Plt (2017) Vol 116 (2): softwre (2017) with the sttisticl pckges Vegn nd Pgirmess. RESULTS AND DISCUSSION After the incution period of seeds of different tree species, 11 gener of fungi (Tle 1) were found, nine gener were detected in untreted seeds nd seven in treted seeds. The numer of fungl colonies developed in non-treted seeds ws higher thn the figure relted to the treted seeds (Tle 1.) The gener of fungi commonly found in the two smples of nlyzed seeds (oth untreted nd treted) comprise Aspergillus spp., Cldosporium spp., Fusrium spp., Penicillium spp., nd Trichoderm spp. However, the numer of these five fungl colonies ws higher in untreted seeds. Despite the occurrence of these fungl gener in seeds sujected to the disinfection with sodium hypochlorite, the use of this compound hs demonstrted negtive impct on the development of the colonies. Similr results were otined y Muniz et l. (2007), who evluted the influence of lortory sepsis in the incidence of pthogens ssocited with seeds nd found tht sepsis influenced seedlings of five forest species in the initil stge. According to these uthors, the ction of sodium hypochlorite in reducing the incidence of fungi ws noticele, indicting tht they re on the surfce of the seed. According to Coutinho et l. (2000), one of the mjor forms of ssocition etween microorgnisms nd seeds is through their loction on the externl tissue, such s pericrp nd tegument, so tht seed tretment with sodium hypochlorite shows efficiency in reducing microorgnisms ssocited superficilly with the seeds. Although the disinfection reduced the occurrence of species nd gener of fungi, we recorded the genus Aspergillus in ll forest species. However, the lrgest numer of fungl colonies of this genus ws found in the untreted seeds (Tle 2). In ddition to the occurrence of higher numer of colonies, this fungl genus showed high vriility. In untreted Myrcrodruon urundeuv seeds, five different species were detected, wheres in treted seeds of such tree three species were identified in smller mounts (Tle 2). The color of colonies llowed us to identify the species Aspergillus niger, A. flvus, A. cndidus, A. ochrceus nd A. glucus, wheres in seeds sumitted to surfce disinfection it ws oserved the occurrence of A. niger, A. flvus, nd A. glucus. The ltter eing the only species with the highest numer of colonies ssocited with the treted Poincinell pyrmidlis seeds, s compred to the untreted ones. According to Círio nd Lim (2003) fungi elonging to the genus Aspergillus spp. re toxigenic, cusing deteriortion in grins nd seeds, nd re lso cosmopolitn sprophytic, esily spred y its light nd dry spores, they my grow in low humidity, fcilitting the development of other genres tht require more moisture. Tle 1. Fungl popultion ssocited with the seed of forestry species from the Cting region sumitted or not to tretment of surfce disinfection. Species nd/or Gener Untreted Seeds Colony (No.) 243 Frequency (%) Colony (No.) Treted Seeds Frequency (%) Aspergillus niger Aspergillus flvus Aspergillus glucus Aspergillus ochrcus Aspergillus cndidus Aspergillus spp Penicillinum spp Tricoderm spp Fusrium spp Alternri spp Chctomium spp Rizoctoni spp Colletotrichum spp Cldosporium spp Curvulri spp Phytophthor spp Totl

4 Do Nscimento et l (2017) Except for Mimos tenuiflor, colonies of Penicillium spp. were found in seeds of ll the other forest species, with the lrgest numer of colonies oserved in untreted seeds of Liidii ferre, followed y Poincinell pyrmidlis nd Myrcrodruon urundeuv (Tle 2). According to Freits et l. (2000), fungi of the gener Aspergillus nd Penicillium re the most found in the seeds during storge, which my impir their qulity, due to its deteriortion. Piñ-Rodrigues nd Vieir (1988) rgue tht the occurrence of the gener Aspergillus, Penicillium, Rhizopus, nd Trichoderm re common in tree seeds, trnsported directly from the plce of collection to the lortory. Although the lrgest numer of colonies of the species or gener of fungi were ssocited with untreted seeds M. urundeuv these hd the lowest numer of occurrence of fungi in seeds sujected to disinfection with M. tenuiflor (Tle 2). After nlyzing the fungl popultion, it ws possile to oserve tht forest species differed significntly (p <0.01) concerning the numer of fungl colonies, the numer of species or gener of fungi, s well s to the numer of germinted seeds for oth the treted seeds nd the untreted ones. Figures 1A nd 1B show tht the verge numer of fungl colonies were smller in the species Mimos tenuiflor, oth for treted nd untreted seeds, differing from the other species. Among ll species nd Fungos fitoptogênicos em sementes d árvore Cting gener detected in this study, Aspergillus niger ws the only species found in the seeds of the forest species Mimos tenuiflor (Tle 2). This result cn e ttriuted to the presence or the effect of compounds with llelopthic ctivities existent in this species which my hve inhiited the development of fungl colonies (Colps et l., 2003). As Silveir et l. (2011) stte, currently Mimos tenuiflor is widely distriuted in the semi-rid region, nd other ntive woody species fil to estlish under its crown, suggesting n llelopthic effect on these species. Studying the tnnin potentil of six tree species from the Brzilin semirid region, Pes et l. (2006) found high potentil of tnnin production in plnts of Mimos tenuiflor. These compounds re produced in the leves of tree species, incresing its concentrtion with ge, mking them less susceptile to the ttck y pthogens (Bezerr, 2008). In ddition to the leves, tnnins my e found in vrious plnt prts, such s the rk, fruits, nd seeds. The most common phenols in plnts re not considered toxic in norml mounts nd conditions, except polymeric phenols referred to s tnnins, which hve the ility to complex nd precipitte proteins from queous solutions (Slunkhe et l., 1990). Sustnces such s these protect plnts from ttck y herivores, invertertes, nd vertertes, y presenting stringent flvor, hindering their intke. Tle 2. Numer of colonies for ech fungl species nd genus ssocited with seeds of four forest species from the Cting region sumitted or not to surfce disinfection. Forest species sujected to disinfesttions M. urundeuv P. pyrmidlis M. tenuiflor L. ferre Species nd Genus Yes No Yes No Yes No Yes No Aspergillus niger Aspergillus flvus Aspergillus glucus Aspergillus ochrceus Aspergillus cndidus Aspergillus spp Penicillium spp Trichoderm spp Fusrium spp Alternri spp Chetomium spp Rizoctoni spp Colletotrichum spp Cldosporium spp Curvulri spp Phytophthor spp Totl numer of colonies

5 Revist de l Fcultd de Agronomí, L Plt (2017) Vol 116 (2): Mimos tenuiflor Liidii ferre Poincinell pyrmidlis Myrcrodruon urundeuv A Avrege of No. of fungl colonies Mimos tenuiflor Liidii ferre Poincinell pyrmidlis Myrcrodruon urundeuv B Avrege of No. of fungl colonies Figure 1. Men numer of fungl colonies from seeds treted (A) nd untreted forest species (B). Mens represented y columns with the sme letter do not differ y Kruskl-Wllis test for multiple comprisons. Mimos tenuiflor is tree species tht cn e useful for griculturl production. According to Brss (2009), the llelopthic effects hve vrious uses tht could contriute to the serch for pesticides, to understnd the ntgonism of syndicted or successive crops; to reduce the use of synthetic hericides, replcing them with llelopthy processes. Similrly to the result of the verge numer of fungl colonies, forest species Mimos tenuiflor presented the lowest vlues s the men numer of species nd gener of fungi, regrdless of the tretment used. Conversely, the highest vlues were found for the species Myrcrodruon urundeuv, even though this species did not differ from Poincinell pyrmidlis e Liidii ferre (Figure 2). Still, the vlues for the untreted seeds were higher (Figure 2B) then treted seeds (Figure 2A), demonstrting tht the use of ctive chlorine s disinfection gent reduces the fungl contmintion, lthough it did not prevent the development of most resistnt colonies, prticulrly those locted internlly in the seeds. These results indicte tht the species Myrcrodruon urundeuv is more susceptile to pthogens nd reinforce the importnce of studies involving the use of other products (plnt extrcts, essentil oils) comined with methods of pthogen control. As pointed out y Silv et l. (2003), seed tretment is n importnt tool to prevent the spred of these pthogens in the field. Another spect nlyzed ws the verge of germinted seeds, which vried mong species, with the highest vlues recorded for the species Poincinell pyrmidlis, sumitted or not to disinfesttion (Figures 3A nd 3B), lthough these species do not differ from Myrcrodruon urundeuv nd Mimos tenuiflor. On the other hnd, the species which showed the lowest vlues in the mens of germinted seeds ws Liidii ferre, oth for the treted nd untreted seeds. In generl, tretment with the surfce disinfection with sodium hypochlorite significntly reduced the incidence of these fungi, regrdless of forest species. Studies conducted with seeds of forest species using seed disinfected with NClO decrese the occurrence of fungi ssocited with the sme (Muniz et l., 2007). Cstellni et l. (1996) oserved tht seed contmintion could ffect in severe form the seed physiologicl qulity nd, in some cses, it completely inhiit the germintion of seeds. However, Oliveir et l. (2006) compred methods of seeds disinfection of golden shower trees (Peltophorum duium) nd detected minly Trichoderm sp., Penicillium sp., Aspergillus niger, nd Fusrium sp. They lso found tht the percentge of seeds infested did not interfere with germintion. According to Crvlho nd Nkgw (2000), the composition of fungl flor lso depends on the wter content of the seed, ecuse chnges in humidity will ffect the chnge in the microflor, oth quntittively nd qulittively. Therefore, the ssocition of fungi with the seeds sometimes will not cuse illness or loss of physiologicl qulity, lthough this ssocition my fvor the prolifertion nd survivl of fungi. Correspondence nlysis shows three distinct res where forest species were seprted due to the distriution of species nd gener of fungi (Figure 4), with the predominnce of the genus Aspergillus which occurred in five different species nd ws detected in four forest species. The tree species tht showed more species nd gener of fungi ws the mstic tree. Mimos tenuiflor, unlike other species, does not pper in the chrt. We ttriute this results to the occurrence of only single colony of fungl species, Aspergillus niger, in the two tretments evluted for this tree species. Using the helth test on filter pper, Cheroini et l. (2008) found in cedr seeds only the following fungi: Aspergillus spp., Chetomium spp., Penicillium spp., nd Trichoderm spp. 245

6 Do Nscimento et l (2017) Fungos fitoptogênicos em sementes d árvore Cting Mimos tenuiflor Liidii ferre Poincinell pyrmidlis Myrcrodruon urundeuv A 0 0,5 1 1,5 2 2,5 3 3,5 4 4,5 5 5,5 6 Averge of No. of fungl species Mimos tenuiflor Liidii ferre Poincinell pyrmidlis Myrcrodruon urundeuv B 0 0,5 1 1,5 2 2,5 3 3,5 4 4,5 5 5,5 6 Averge of No. of fungl species Figure 2. Averge numer of fungl species/gener in forest species seeds which were sujected to tretment (A) nd which were not sujected to tretment (B). Mens represented y the columns of the sme lowercse letter do not differ from one nother y Kruskl-Wllis multiple comprison tests. Mimos tenuiflor Liidii ferre Poincinell pyrmidlis Myrcrodruon urundeuv c A Mens of No. of germinted seeds Mimos tenuiflor B Liidii ferre Poincinell pyrmidlis Myrcrodroun urundeuv Mens of No. of germinted seeds Figure 3. Mens of germinted seeds mong forest species seeds which were sujected to tretment (A) nd which were not sujected to tretment (B). Mens represented y the columns of the sme lowercse letter do not differ from one nother y Kruskl-Wllis multiple comprison tests. Oliveir et l. (2012) verified tht prior disinfection with 1.5% sodium hypochlorite for five minutes reduces fungl contmintion, fcilitting the evlution of the helth test, nd the compound does not ffect seedling emergence, eing eneficil for the estlishment of the seedling Schizoloium mzonicum. According to Ruiz-Filho et l. (2004), fungi such s Chetomium, Trichoderm, Penicillium, Aspergillus, nd Rhizopus re ssocited with seed deteriortion nd its ction is dependent on their physicl nd physiologicl conditions, t the eginning of storge, nd on the dominnt environmentl fctors during this period. 246

7 Revist de l Fcultd de Agronomí, L Plt (2017) Vol 116 (2): A B Figure 4. Fungl species identified in forest species in treted (A) nd untreted (B) seeds. Aspergillus niger (An); Aspergillus glucus (Ag); Aspergillus ochrcceus (Ao); Aspergillus cndidus (Ac); Aspergillus flvus (Af); Aspergillus sp (Asp); Penicillium (P); Fusrium (F); Colletotrichum (Cl); Cldosporium (Cl); Curvulri (Cv); Phytophthor (Phy); Trichoderm (T); Cldosporium spp. (Clsp); Rhizoctoni (R); Chetomium (Ch), Numers of unidentified fungl species (N) In Figure 4, it cn e oserved tht the species Myrcrodruon urundeuv presented the highest microflor ssocited with untreted seeds relted to treted seeds of the remining species. The incidences of the fungi Aspergillus niger (An), Aspergillus glucus (Ag), Penicillium sp (P), Fusrium sp. (F), Trichoderm sp. (T), Aspergillus flvus (Af), nd Cldosporium sp. (Cl) ws significntly reduced in the treted seeds, indicting tht they were locted in the externl prt of the seeds or dispores of Myrcrodruon urundeuv (Allemão), Liidii ferre, nd Poincinell pyrmidlis (Tul.), nd tht the use of sodium hypochlorite ws efficient. According to Botelho et l. (2008), the disinfection with 1% sodium hypochlorite for 3 minutes reduced significntly the incidence of the fungi Aspergillus spp., Curvulri sp. nd Trichothecium sp. ssocited with seeds of yellow nd purple trumpet trees, in lmost ll smples. CONCLUSION The gener of fungi commonly found in the seeds of the studied tree species were: Aspergillus, Cldosporium, Fusrium, Penicillium, nd Trichoderm. The incidences of fungi on seeds of the studied tree species were reduced y the disinfection using 2% sodium hypochlorite. The genus Aspergillus presented the highest numer of colonies nd diversity of fungl species in treted nd untreted seeds, eing present in ll tree species. The seeds Mimos tenuiflor hve the lowest fungl incidence. 247 REFERENCES Alfens, A.C. & R.G. Mfi Métodos em Fitoptologi. Ed. UFV, p.382. Botelho, L. S., M.H.D. Mores & J.O.M. Menten Fungos ssocidos às sementes de ipê-mrelo (Teui serrtifoli) e ipê-roxo (Teui impetiginos): incidênci, efeitos n germinção e trnsmissão pr s plântuls. Summ Phytopthologic, Botuctu, 34: 4: Brnett, H.L. & B.B. Hunter. Illustrted gener of imperfect fungi. 4. ed. St. Pul, MN: Amicn Phytoptologicl Society Press, p. Brss, F.E.B Análise de tividde lelopátic de extrto quoso de flsmurt sore germinção de picão-preto e cruru. Enciclopédi Biosfer, 8:1-19. Bezerr, D.A.C Estudo Fitoquímico, Bromtológico e Microiológico de Mimos tenuiflor (Wild) Poiret e Piptdeni stipulce (Benth) Ducke. Dissertção presentd à Universidde Federl de Cmpin Grnde o Progrm de Pós-Grdução em Zootecni, áre de concentrção em Sistems Agrossilvipstoris no Semiárido, Ptos. Crvlho, N.M. & J. Nkgw Sementes: ciêncis, tecnologi e produção. 4 ed, FUNEP, pp.588. Colps, F.T., E.O. Ono, J.D. Rodrigues & J.R.S. Pssos Effects of Some Phenolic Compounds on Soyen Seed Germintion nd on Seed-orne Fungi. Brzilin Archives of Biology nd Technology n Interntionl Journl (46)2: Cstellni, E.D., A. Silv, M. Brreto & I.B. Aguir Influênci do trtmento químico n populção de fungos e n germinção de sementes de Buhini

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