The present experiment was aimed to evaluate the response of sugarcane varieties inoculated with endophytic diazotrophic bacteria. Sixteen sugarcane varieties were used for three kind of treatrnents (no inoculation and inoculation with 120 kg N/hectare). The inoculant was a mixture of five strains of bacterial species isolated from sugarcane. There were significant differences between genotypes (G) and genotype x treatment interaction (GxT) for ali the traits. The variety SP 71-6949,SP 78-4764and RB 867515 were found responsive to biological nitrogen fixation and can be used in breeding programs which are directed to study and selection of promising genotypes responsible for the biological nitrogen fixation

AGUIAR, M. S. de

AMARAL, A. L. do

CAMARA, T. M. M.

IVO, W. M. P. de M.

MARAFON, A. C.

MORAIS, L. K. de

RAMOS, N. P.

REIS, V. M.

SILVA, P. de A.

RCAAP - Repositório Científico de Acesso Aberto de Portugal

Evaluation of performance of sugarcane genotypes inoculated with endophytic diazotropic bacteria.

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Balancing Sugar and Energy Production in Developing Countries: Sustainable Technologies and Marketing Strategies, New Delhi, India, pp 92-95 (2011)EVALUATION OF PERFORMANCE OF SUGARCANE GENOTYPES INOCULATEDWITH ENDOPHYTIC DIAZOTROPHIC BACTERIALlZZ KEZZY DE MORAIS1, PAULO DEALBUQUERQUE E SI LVA1, VERÔNICAMASSENAREIS2,MARCELO SFEIR DE AGUIAR1, TASSIANO MAXWELL MARINHO CÂMARA 1, ANDERSON CARLOSMARAFON1, WALANE MARIA DE MELLO IV01, ADRIANE LEITE DO AMARAL 1ANONILZA PATRíCIA RAMOS3lEmbrapa Tabuleiros Costeiros, Rio Largo, Alagoas, Brazil2Embrapa Agrobiologia, Rio de Janeiro, Brazil.3Embrapa Meio Ambiente, São Paulo, Brazil.AbstractThe present experiment was aimed to evaluate the response of sugarcane varieties inoculated with endophyticdiazotrophic bacteria. Sixteen sugarcane varieties were used for three kind of treatrnents (no inoculation andinoculation with 120 kg N/hectare). The inoculant was a mixture of five strains of bacterial species isolated fromsugarcane. There were significant differences between genotypes (G) and genotype x treatment interaction (GxT)for ali the traits. The variety SP 71-6949,SP 78-4764and RB 867515were found responsive to biological nitrogenfixation and can be used in breeding programs which are directed to study and selection of promising genotypesresponsible for the biological nitrogen fixation.Key words: Biological nitrogen fixation; sugarcane; genotypic performance.INTRODUCTIONBrazil produces about 624.99 million tons of sugarcane,of which 336.2 million tons (53.8%) utilized in theproduction of 27.7 billion of liters of ethanol and theremaining, 288.7 million tons (46.2%) was used toproduce sugar (CONAB 2010). The sugarcane is the mainsource of energy (biofuel). The great advantage ofsugarcane for the production of biofuel is the positiveenergetic balance. In Brazil this balance is approximately+9, which means .for every unit of energy consumed inthe production process, nine units are generated(Macedo and Koller 1997). This highly positive balancecan be attributed to the cultural yields with low nitrogenfertilization (Urquiaga et al. 1992). In other productivecountries like USA, Australia, Cuba, Venezuela andPeru, the nitrogen fertilization varies between 200 to 400kg/ha (Xavier 2006; Boddey et alo1995), resulting in anenergetic balance that rarely achieves + 1 (Azeredo et al.1986).The sugarcane improvement in Brazil have resulted inlow nitrogen fertilizer inputs of about 60 kg/ha becauseof genotypes that rarely respond sigrrificantly to nitrogennutrient (Azeredo et al. 1986). These types of studieshelp to observe that there might be some genotypes whichexpress their potential for the biological nitrogen fixation(BNF) associated with diazothrophic bacteria. Manyauthors have studied different genotypes in the presenceof diazothrophic bacteria which are responsible for thebiological nitrogen fixation in sugarcane (Boddey et al.2001; Xavier 2002; Baldani et alo2002; Boddey et alo2003;Baldani and Baldani 2005, Nogueira et al. 2005; Oliveiraet alo2006). However, various factors can be involved inthe efficiency of the biological nitrogen fixation in thesugarcane culture. Because its a biological process whichdepends on the interaction between the microorganisms,plants and environment. To express the fixationpotential, it is necessary that all these three componentsbe in good conditions, or the microorganisms involvedbe genetically favorable for this interaction and thegenotypes be responsive to the inoculation and theenvironrnent. The objective of this work was to evaluatethe growth characteristics of different sugarcanevarieties subjected to the inoculation with endophyticdiazotrophic bacteria.MATERIAL AND METHODSThe experiment was conducted during the cropping yearof 2010/2011 in the farm area of Triunfo Mill, Atalaia,State of Alagoas (Brazil). The experiment was conductedin randomized block design with three replications in afactorial scheme (16 x 3) comprized of total 48 treatrnents.The treatments were composed of 16 cultivar incombination with inoculant, without inoculant andwith nitrogenous fertilization (120 kg of Nitrogen). Everyplot had six rows of 5 m separated by 1.40 m row to rowspace. The fertilizer, excepting nitrogen, was appliedaccording to soil analysis. The mixture of endophyticdiazotrophic bacteria (inoculants) supplied by EmbrapaAgrobiology, was used before the planting. Theinoculants contained a mixture of bacteria, viz.,Corresponding author: Lizz Kezzy de Morais. e-mail: lizz@cpatc.embrapa.brLizz Kezzy de Morais et aiGluconacetobacter diazotrophicus, Herbaspirillumseropedicae, H. rubrisubalbicans, Azospirillum amazonensee Burkholderia tropica. Microbial inoculants was mixedwith 100 liters of water and the setts used in plantingwere immersed in this mixture for a period of 1 h andplanted immediately. After seven and nine months ofplanting, the field evaluations were made to record thefollowing data: the average stalk height; average staIkdiameter (at the height of 1/3 from the base with thehelp of a paquímeter); leaf length; leaf width; number ofdried leaves; The number green leaves and Chlorophyllcontent (determined by indirect method using SPAD 502Chlorophyll meter machine, Minolta Corporation,Ramsey, Japan). The averages values were obtained from10 replicates per leaf (+3 leaf). The statistical analyseswere performed by using Genes ProgramoRESULTS AND DISCUSSIONSignificant differences (p <0.001) were observedamongst genotypes and treatments which indicated thepresence and genetic variability for all traits. Nosignificant difference observed between seasons (Table1). This variability reflected the heterogeneity of thegenetic material, and indicated the possibility ofidentifying promising genotypes responsible for thebiological nitrogen fixation. The variation coefficient(VC) ranged from from 6.33 to 14.66, and showed goodexperimental precision in the determination of growthcharacteristics as in plant height (PH), diameter of stem(DS), leaf length (LL),width of leaf (WL), number of driedleaves (NDL), number of green leaves (NGL), andchlorophyll content (CC). Table 2 showed the estimatedaverage of genotypes in each treatment. The varieties 1,2 and 7 inoculated with endophytic diazotrophicbacteria showed a higher average when compared tothe same variety with other treatments (non-inoculatedand nitrogen) for these characteristics, this reflected theresponsive behavior of these genotypes to the biologicalfixation of nitrogen. Varieties 10, 14 and 15 showed thebest performance and the highest values for the sameTable 1.93characteristics, and varieties 15 and 16 showed gooddevelopment in growth when they were not subjected totreatment of inoculation and nitrogen application.According to the reports of Ferreira at aI. (2007), DS andPH features are positively correlated with productivityand tons per hectare (TCH). Plants with high values ofheight and diameter can be suggested as productiveplants. The varieties 1, 2 and 7 subjected to inoculationshowed high mean values of plant height and diameterof the stem. LL, DS and PH features were linked to thegrowth, and the development of the plants was directlyrelated to the stimulus provided by plant hormones.Several diazotrophic bacteria are able to producephytohormones responsible for the observed stimulatoryeffect on plant growth. Bastian et aI. (1998) detected thepresence of IAA and gibberellins AI and A3 ín culturesof Herbaspirillum seropedicae. Similar results wereobtained by Fuentes-Ramirez et aI. (1993) evaluating theIAA production by Gluconacetobacter diazotrophicusstrains. Differences between sugarcane varieties and thepotential for nitrogen fixation were studied by Coelho etai. (2003), who reported that the BNF was observed inall commercial sugarcane genotypes evaluated(RB739735, SP79-2313, RB72454, RB758540, RB835089,RB825336 and SP79-2312), and also in wild accessions,Krakatau (5 . spontaneum) and Chunnee (5. barberi). Inthis study, the greatest contributions of BNF wereobserved in varieties RB739735, RB758540, RB835089and SP79-2312 varieties. Boddey et ai. (2003) stated thatthe capacity for N2 fixation greatly depend on thesugarcane variety. In the study conducted by Polidoroet aI. (2001), the RB72454 and SP80-1842 commercialsugarcane varieties showed high potential for BNF.Therefore, accounting for the N from biological nitrogenfixation and the differential response in each particulargenotype in plant breeding programs is still an issuewhich should be thoroughly studied especially withregards to the sugarcane culture. Such studies areimportant to understand the genotype, environmentaland microorganism interaction, because bacteria presentAnalysis of variance for seven characteristics evaluated [Plant height (PH); Diameter of the stem (DS);Length of the leaf (LL); Width of the leaf (WL); Number of dead leaves (NDL); Number of live leaves (NLL)and Chlorophyll content (CC)] at the seventh and ninth months of age.VF DS (dm) CTDF PH (em)Genotype (G)Season (S)Treatment (T)GxSGxTSxTGxSxTResíduoAverageCoefficient of Variation (%)151215302301802361.79**226499.55**224.03NS134.43NS244.41*111.58NS49.87NS139.71119.959.8534.49**30.65NS5.65NS2.14NS4.87*0.28NSl.72NS3.1426.226.76MSLL (em) WL (em) NDL NLL1272.41**272.87NS268.55NS141.84**134.12**14.14NS34.53NS58.49120.756.3394.20**17732.43**5.32NS66.86**20.77*44.69NS16.45NS13.5334.4610.671.61**56.94**1.02NS0.33**0.14NSO.01NS0.13NS0.133.3311.156.12**932.13**3.04NS3.31*2.27NS1.32NS0.81NS1.688.8614.661.78**84.95**0.47NS0.77*0.45NS0.14NSO.23NS0.385.1612.0194 Evaluation of perfarmance of sugarcane genatypes inoculated with endaphytic diazotrophic bacteriaTable 2. Estimated measurements of 16 sugarcane genotypes referring to the eharacteristics of Plant height (PH);Diameter of the stem (DS); Length of the leaf (LL); Width of the leaf (WL); Number of dead leaves (NDL);Number of live leaves (NLL) and Chlorophyll content (CC) evaluated at the seventh and ninth months ofageGenotypel/ Treatament PH (em) DS (dm) LL (em) WL (em) NDL NLL cr1 Not-inoeulation 127.25 25.11 117.62 3.64 9.16 5.25 33.071 Inoeulation 143.25 26.11 117.58 3.81 10.45 5.58 33.361 Nitrogen 127.33 25.14 104.58 4.16 8.04 5.25 32.712 Not-ínoculation 137.83 24.75 120.00 3.24 8.91 5.29 35.332 lnoeulation 150.37 25.11 126.08 3.31 8.91 5.79 37.222 Nitrogen 126.33 25.00 116.87 3.38 9.08 4.87 33.783 Not-inoculation 118.91 29.43 127.61 3.89 9.36 4.65 34.243 Inoculation 112.87 27.81 128.21 3.32 7.91 5.37 35.033 Nitrogen 101.54 27.28 120.16 3.67 8.37 4.62 37.794 Not-inoculation 137.41 24.82 127.44 3.20 9.72 5.11 36.524 Inoeulation 125.87 26.48 128.37 3.26 8.83 5.12 37.094 Nitrogen 132.79 24.94 125.50 3.28 8.87 4.91 35.965 Not-ínoculation 104.41 24.32 119.08 2.68 9.20 4.91 36.945 Inoeulation 100.47 25.40 109.87 2.66 9.77 5.04 35.775 Nitrogen 114.21 25.64 121.50 3.14 8.54 4.58 33.946 Not-inoculatíon 107.83 24.70 111.08 3.24 9.33 4.83 36.556 Inoeulation 99.95 23.71 110.41 3.33 9.20 5.37 38.826 Nitrogen 105.91 25.46 106.50 3.73 9.95 4.71 35.867 Not-inoeulation 129.58 27.05 127.79 3.52 9.37 5.04 32.077 Inoeulation 141.12 25.36 134.08 3.42 9.54 5.04 33.757 Nitrogen 139.21 25.90 128.29 3.77 8.87 4.87 32.138 Not-inoeulation 125.33 27.91 117.04 3.32 9.54 6.15 34.228 Inoeulation 124.33 29.47 115.37 3.44 9.41 5.91 32.658 Nitrogen 128.41 27.95 114.33 3.47 8.91 5.62 32.839 Not-inoculation 130.58 23.57 121.16 3.74 8.87 5.20 30.719 Inoeulation 123.54 25.03 110.91 3.40 9.16 4.87 33.219 Nitrogen 122.33 24.81 111.62 3.52 9.00 5.37 30.2510 Not-inoculation 117.83 25.63 135.58 3.27 10.25 5.37 38.4210 Inoeulation 109.51 24.66 133.76 3.17 9.50 4.70 37.1410 Nitrogen 111.00 26.40 130.12 3.53 9.16 5.29 41.2811 Not-ínoculation 109.33 26.49 114.00 2.61 7.83 5.58 38.4211 Inoeulation 103.87 28.65 111.12 2.73 8.45 5.34 33.7511 Nitrogen 112.62 26.77 120.50 2.77 8.21 5.87 35.8812 Not-inoculation 124.91 26.69 116.58 3.05 8.66 5.66 31.8412 lnoeulation 119.70 28.37 105.37 2.80 7.50 5.16 29.6212 Nitrogen 120.83 26.99 110.16 2.95 7.87 5.08 28.7213 Not-inoculation 114.45 23.15 133.87 2.98 8.71 4.62 31.7213 Inoeulation 111.33 25.47 119.25 3.29 8.37 5.04 29.1213 Nitrogen 124.16 25.58 134.08 3.49 8.83 5.08 36.6114 Not-inoculatíon 115.75 25.46 142.12 3.31 8.08 4.87 35.2314 Inoeulation 105.45 27.19 137.25 3.31 5.91 5.16 32.8314 Nitrogen 114.95 26.07 131.46 3.49 8.37 4.91 34.2015 Not-inoculation 133.37 29.01 117.70 3.67 8.87 5.62 34.1815 lnoculation 120.54 26.89 114.29 3.35 9.12 5.25 35.2915 Nitrogen 128.41 27.04 120.33 3.46 9.29 5.55 31.6616 Not-inoculation 109.21 28.34 114.29 3.39 9.21 4.79 35.7316 Inoeulation 102.91 28.19 114.12 3.38 7.91 4.62 33.0216 Nitrogen 108.41 27.45 121.21 3.33 8.95 4.66 37.541/ Sugarcane varieties: 1- SP 71-6949; 2-SP 78-4764; 3-SP 79-1011; 4-SP 81-3250; 5-RB 72 454; 6-RB 845210; 7-RB 867515; 8-RB863129;9-RB 92579; 10-RB 93509; 11-RB 931003; 12-RB 951541; 13-RB 98710; 14-RB 98395; 15-VAT 90-186; 16-VAT 90-212.Lizz Kezzy de Morais et aiin the inoculant have a direct influence on the growthcomponents and are directly related to sugarcaneproductivity.CONCLUSIONSThe sugarcane varieties SP 71-6949, SP 78-4764 and RB867 515were responsible for the biological nitrogenfixation and may be utilized in breeding programs ma deto the selection of promising genotypes fixing biologicalnitrogen.REFERENCESAzerdo DF, Bolsanello J, Weber H, Vieira JR (1986)Nitrogênioem cana-planta - doses e fracionamento. 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Evaluation of performance of sugarcane genotypes inoculated with endophytic diazotropic bacteria.

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