Este trabalho de investigação refere-se ao uso do reator anaeróbio híbrido para tratamento de esgoto sanitário, com configuração baseada no reator anaeróbio de manta de lodo (UASB) com inclusão de: meio suporte sobre as calhas de coleta de gás (denominado reator anaeróbio híbrido - UAHB) e, também, meio suporte na zona de reação (denominado reator anaeróbio híbrido modificado - UAHBmod). Para o desenvolvimento desta pesquisa, no Campus I da USP de São Carlos-SP foram construídos dois reatores experimentais de 18,8 m3 cada: um reator UASB, com função de controle, e um reator UAHB. Primeiramente os reatores foram operados por período de 200 dias, com tempo de detenção hidráulica (TDH) de 6 h. Após serem inoculados, com 80 dias de operação, os reatores atingiram o estado de equilíbrio dinâmico aparente, com geração de alcalinidade, baixa concentração de ácidos voláteis e eficiência de remoção média de DQO, de 84% e 85% e de DBO de 87% e 91%, respectivamente para o UASB e o UAHB. Após esse período, os reatores foram submetidos a aumento da velocidade ascensional (Vasc) (mediante recirculação do efluente) de 0,78 m.h-1; 1,17 m.h-1; 1,56 m.h-1 e de 1,96 m.h-1. O UAHB mostrou ser menos susceptível ao aumento da Vasc do que o UASB. Além da análise da operação dos reatores, foram realizados os ensaios hidrodinâmicos e avaliada a estrutura da comunidade microbiana, por microscopia ótica, epifluorescência e pela técnica do DGGE. Após esse período preliminar, os reatores UAHB e UAHBmod, operados com TDH de 6h e Vasc de 0,78 m.h-1, atingiram o estado de equilíbrio dinâmico aparente, com geração de alcalinidade, baixa concentração de ácidos voláteis e eficiência de remoção média da matéria orgânica, de 71% e 76% em DQO, e de 72% e 87% em DBO, respectivamente para o UAHB e UAHBmod. Após este período, o reator UAHBmod, submetido a Vasc de 1,56 m.h-1, promoveu remoção de 74% de DQO, e de 87% de DBO. / This research refers to the use of a hybrid anaerobic reactor (UAHB) for domestic wastewater treatment. The configuration of this reactor is based on a sludge bed anaerobic reactor (UASB); in the first instance, a media support above the gas collection apparatus (also known as hybrid anaerobic reactor) was provided and later, a media support on the reaction zone (also known as hybrid modified anaerobic reactor - UAHBmod) was provided. Two reactors, with a volume of 18.8 m3, each, were built for this research at Campus I, USP in São Carlos - SP-Brazil. One UASB reactor acted as a control, and the other as a UAHB reactor. In the preliminary essays, the reactors were operated with 6h of hydraulic detention time (HDT) for 200 days. After inoculation, the reactors attained the apparent dynamic equilibrium state after 80 days of operation, with alkalinity generation, low volatile acids concentration and mean organic matter removal of 84% and 85% in terms of COD, and 87% and 91% in terms of BOD, for UASB and UAHB reactors, respectively. After this period, the reactors were submitted to an increasing in up velocity (Vup) of 0.78 m.h-1; 1.17 m.h-1; 1.56 m.h-1 and 1.96 m.h-1. The UAHB reactor showed lesser susceptibility for Vup increase than the UASB reactor. Hydrodynamic tests were also done on the reactors, in addition to routine operational analysis. The structure of the microbial community was evaluated by optical and epifluorescence microscopy, and the DGGE technique. After this step, the UAHB and the UAHBmod reactors were operated out 6h of HDT and Vup of 0.78 m.h-1. The reactors attained the apparent dynamic equilibrium state with alkalinity generation, low volatile acids concentration and mean organic matter removal of 71% and 76% in terms of COD, and 72% and 87% in terms of BOD for the UASB and UAHBmod reactors, respectively. After this period, the UAHBmod reactor was subjected to a Vup of 1.56 m.h-1 and achieved removal efficiencies of 74% COD and 87% BOD.

Diferentes métodos de caracterização e inferências filogenéticas baseadas na seqüência de alguns genes nucleares indicam que Typanosoma cruzi pode ser dividido em dois grupos principais, denominados T. cruzi I e T. cruzi II. Outros subgrupos de isolados foram descritos, tais como grupo de rDNA 1/2 e zimodema 3 (Z3) cuja relação filogenética com os grupos T.cruzi I e T.cruzi II ainda não está clara. O cariótipo molecular é uma ferramenta que permite abordar diferentes aspectos do genoma de organismos. Neste trabalho, caracterizamos o cariótipo molecular de 22 isolados de T. cruzi a partir da separação de seus cromossomos por eletroforese de campo pulsado e hibridação com sondas que representam genes codificadores de proteína e de RNA. Verificamos extenso polimorfismo cromossômico entre os isolados e que isolados pertencentes aos grupos T. cruzi II, rDNA 1/2 e Z3 têm cromossomos de tamanho molecular maior (até 3,5 Mb) em relação a isolados de T. cruzi I (até 2,8 Mb). Os dados do cariótipo molecular também foram utilizados para averiguar o significado evolutivo do polimorfismo cromossômico. As análises fenéticas foram baseadas no índice de diferença absoluta de tamanho cromossômico (aCSDI) obtido a partir da hibridação dos cromossomos com um número variável de marcadores genéticos. Inicialmente analisamos nove isolados, classificados por diferentes abordagens moleculares nos gruposT. cruzi I, T. cruzi II e grupo de rDNA 1/2, este último considerado um grupo de isolados híbridos e incluído por alguns autores no grupo T. cruzi II. Dendrogramas de aCSDI obtidos a partir de 3 a 21 sondas definiram em todos os casos três grupos: dois correspondentes a T. cruzi I e T. cruzi II e um terceiro, ao grupo de rDNA 1/2. O isolado CL Brener - organismo de referência do Projeto Genoma deT. cruzi - ora agrupou com T. cruzi II ora com o grupo de rDNA 1/2, ilustrando seu caráter híbrido. Três grupos também foram observados no dendrograma construído com dados de polimorfismo de tamanho de fragmentos de restrição (RFLP) hibridados com dezoito sondas. A topologia dos dendrogramas de dados de cariótipo molecular e de RFLP é similar, com um coeficiente de correlação significante (r = 0.86062; P < 0.0001), corroborando uma forte estruturação dos grupos. Subseqüentemente, avaliamos a posição de isolados de Z3 em relação aos três grupos acima mencionados, uma vez que alguns autores situam Z3 no grupo T. cruzi II. Analisamos o padrão de hibridação de 9 sondas com os cromossomos de 19 isolados (oito de Z3, três de T. cruzi I, três de T. cruzi II e cinco de rDNA 1/2). A topologia dos dendrogramas mostrou quatro grupos correspondentes, respectivamente, a T. cruzi I, T. cruzi II, grupo de rDNA 1/2 + CL Brener e Z3 (oito dos nove isolados). Este estudo também revelou que isolados híbridos têm uma maior proporção de cromossomos homólogos de tamanhos diferentes em comparação com isolados não híbridos. De um modo geral, nossos resultados mostram que cromossomos são caracteres valiosos para a identificação de táxons em T. cruzi. Uma vez que isolados do grupo de rDNA 1/2 apresentam cistrons ribossômicos de tipo 1 e de tipo 2, caracterizamos a distribuição dos cistrons nas bandas cromossômicas destes isolados. Verificamos que a fração majoritária dos genes de rDNA do tipo 2 está localizada na banda de 1,5 Mb e que os cistrons do tipo 1 estão localizados principalmente na banda de 1,1 Mb. Também estimamos o número de cromossomos e tamanho do genoma de quatro isolados de T. cruzi , com base na análise densitométrica da intensidade de fluorescência dos cromossomos corados com SYBR Green I. Para esta finalidade idealizamos um modelo matemático que permitiu estimar 52, 65, 72 e 44 cromossomos (na célula 2N), respectivamente, para CL Brener, Esmeraldo cl3, SO3 cl5 e Silvio X10 cl1. O tamanho do genoma dos isolados foi calculado, respectivamente em, 70 Mb, 78 Mb, 94,7 Mb e 47 Mb. Os novos aspectos do cariótipo molecular de T. cruzi aqui apresentados contribuirão para a compreensão da organização do genoma deste parasita e auxiliarão na atribuição de arcabouços de genes (scaffolds) aos cromossomos de CL Brener. / Different typing methods and phylogenetic inference based on the nucleotide sequence of few nuclear genes indicate that Trypanosoma cruzi can be divided into two major groups named as T. cruzi I and T. cruzi II. Additional subgroups of isolates have been described, such as group of rDNA 1/2 and zymodeme 3 (Z3), whose phylogenetic relationships with the T. cruzi I and T. cruzi II groups is not clear. The molecular karyotype is a tool that allows investigation of particular aspects of the genome of organisms. In this study, we have characterized the molecular karyotype of 22 isolates following the separation of chromosomes by pulsed-field gel electrophoresis and hybridization with probes that represent genes coding for proteins or RNA species. We observed an extensive chromosome polymorphism between the isolates and that isolates typed as T. cruzi II, rDNA 1/2 and Z3 have chromosome of larger molecular size (up to 3.5 Mb) in relation to T. cruzi I isolates (up to 2.8 Mb). Data from molecular karyotype have been also used to verify the evolutionary meaning of chromosome polymorphism. The phenetic analyses were based on the absolute chromosomal size difference index (aCSDI), calculated from the hybridization of a variable number of genetic markers. Initially, we analyzed nine isolates, classified by different molecular approaches into T. cruzi I,T. cruzi II and rDNA 1/2 groups. This latter group has been considered of hybrid genotypes and has been included by some authors in the T. cruzi II group. aCSDI-based dendrograms obtained from 3 to 21 probes defined in all the cases three clusters: two corresponding, respectively, to T. cruzi I and T. cruzi II groups; and a third one, to rDNA group 1/2. CL Brener - the reference organism of the T. cruzi Genome Project - was alternatively positioned in T. cruzi II or rDNA 1/2 group clusters, illustrating the hybrid nature of this isolate. Three clusters were also observed in the dendrogram constructed with restriction fragment length polymorphism (RFLP) data from 18 probes. The topology of the chromosome and RFLP dendrograms is similar, with a significant correlation coefficient (r = 0.86062; P < 0.0001), supporting a strong structuring of the clusters. Subsequently we evaluated the position of Z3 isolates in relation to the above-mentioned groups, because some authors clustered Z3 with T. cruzi II group. For this purpose we determined the hybridization pattern of 9 probes with the chromosomes of 19 isolates (eight of Z3; three of T. cruzi I; three of T. cruzi II and five of rDNA 1/2). The topology of the aCSDI-based dendrograms showed four clusters corresponding, respectively, to T. cruzi I, T. cruzi II, rDNA 1/2 + CL Brener and Zymodeme 3 (eight out of nine isolates). This study also revealed that hybrid stocks have a larger proportion of two different-sized homologous chromosomes, as compared with non-hybrid. Overall, our results show that chromosomes are valuable characters for identification of evolutionary groups in T. cruzi. Because rDNA 1/2 isolates have ribosomal cistrons of type 1 and type 2, we have characterized the distribution of both cistrons in the chromosomal bands of these isolates. We verified that the majority of type-2 rDNA genes are localized in a 1.5 Mb band, whereas type-1 cistrons are mostly concentrated in a 1.1 Mb band. We have also estimated the number of chromosomes and genome size of four T. cruzi isolates, based on the densitometric analysis of the fluorescence intensity of chromosomes stained with SYBR Green I. For this purpose, we devised a mathematical model that allowed estimating 52, 65, 72 and 44 chromosomes (2N cell), respectively, in CL Brener, Esmeraldo cl3, SO3 cl5 and Silvio X10 cl1. The genome size in these isolates has been calculated, respectively, as 70 Mb, 78 Mb, 94,7 Mb and 47 Mb. The novel aspects of T. cruzi karyotype here presented contribute to the comprehension of the genome organization of this parasite and will assist the assignment of scaffold to the CL Brener chromosomes.

O trabalho teve por objetivo avaliar fatores associados à ensilagem da cana-deaçúcar, com destaque para a aplicação de aditivos químicos e microbianos sobre a dinâmica fermentativa, composição bromatológica, atividade da álcool desidrogenase e desenvolvimento e diversidade da micloflora em silagens de cana-de-açúcar. No ensaio conduzido durante 110 dias o delineamento experimental adotado foi o inteiramente ao acaso, com 4 tratamentos, 2 repetições, e seis épocas de abertura (1, 3, 7, 15, 35, 110 dias). Os tratamentos foram: uréia 1% MV e os inoculantes microbianos Lactobacillus buchneri (3,65x105 ufc/g da MV) e a combinação de bactérias Pedioccocus pentosassus e Lactobacillus buchneri (1x106 ufc/g MV). As maiores variações na composição bromatológica e perdas de MS, das silagens controle ocorreram dos 7 aos 15 dias, estabilizando após esse período. As regressões ajustadas para perdas de MS e carboidratos solúveis foram bem similares e de forma contrária ao acúmulo de FDN. As perdas por gases alcançaram valores de 28,27%, de carboidratos solúveis em apenas 2,98% e FDN em torno de 67,77% da MS. Os aumentos nos teores de etanol e perda na digestibilidade nas silagens controle se extenderam até o 35º dia, com valores máximos de etanol de 12,23%. Foi possível relacionar etanol com a digestibilidade mostrando que cada 1% de aumento nos teores de etanol, 2 unidades de digestibilidade foram perdidas. Os aditivos uréia e o aditivo Lactobacillus buchneri mais Pediococcus foram eficazes em diminuir a produção de etanol (2,75 e 1,30 vs 8,27% no tratamento controle), em diminuir perdas de MS em 47 e 60%, e de carboidratos soluveis em 22 e 56% em relação à silagem controle, respectivamente. As silagens aditivadas com uréia obtiveram maiores valores de pH e maiores valores de ácido lático em relação às silagen controle. As silagens aditivadas com L. buchneri apenas foram as de maiores produções de etanol, acima da silagem controle (11.53 vs 8.27%), além de grandes perdas de matéria seca e baixa digestibilidade pelo acúmulo de FDN, comparáveis às silagens controle. A diferença entre aditivos na composição químico-bromatológica e perdas ocorreu após 7 dias de fermentação. Os dados apresentados pelos aditivos uréia e L. buchneri mais Pediococcus foram ajustados em curvas simples, através de modelos lineares, para descrever e predizer as variações durante a ensilagem. Os tratamentos controle e a aditivação com L. buchneri apenas, pelas altas taxas fermentativas, observaram melhor ajuste dos dados em polinômios de segundo e terceiro grau. Apesar dos altos teores de ácido acético em todas as silagens, principalmente nas silagens aditivadas com a combinação de bactérias, não foram verificadas efeitos deste sobre a população de leveduras. Os teores obtidos de ácido lático e ácido propiônico e a relação entre esses ácidos e o ácido acético, durante a fermentação, conseguiu explicar parte do sucesso dos tratamentos uréia e L. buchneri mais Pediococcus na redução da atividade da enzima álcool desidrogenase e na producão de etanol. A análise de grupamentos hieráquicos mostrou que os aditivos alteraram a diversidade bacteriana durante a ensilagem. / The present trial aimed to study the ensiling associated factors of sugarcane focusing on chemical and microbial additives on fermentation, chemical composition, enzymatic activity of alcohol dehydrogenase and the microflora development and diversity in sugarcane silages. A complete randomized design was set to a 110-d trial with 4 treatments, two replications within 6 opening dates (1, 3, 7, 15, 35, 110-d). Treatments were described as follows: urea 1% (wet basis), Lactobacillus buchneri (3.65x105 cfu/g of forage), a combination of Pediococus pentosassus and Lactobacillus buchneri (1x106). Major variation observed on the chemical composition and the DM losses in sugarcane silages without additives took place from the day 7 through the day 15. Losses of DM and soluble carbohydrates showed similar trend and in opposition to the NDF increase. Gases losses averaged 28.27%, while the soluble carbohydrates and NDF contents reached respectively, 2.98% and 67.77% when fermentation was stabilized. Conversely, ethanol and the digestibility were changed across the storage period up to the day 35, with ethanol content increasing to 12.23%. 1% of ethanol increase was associated with 2 percentage units of digestibility decrease. Both urea and the combination of microorganisms were effective in decrease the ethanol content (2.75, 1.30 vs 8.27% - without additives), decrease DM losses (47 and 60%) and reduce soluble carbohydrates losses (22 and 56%) when compared to the control treatment. The urea treated silages showed higher pH and lactic acid values. The L. buchneri treatment led to higher ethanol content (11.23 vs 8.27%) compared to the control, resulting in low DM recovery rate, higher losses and decreased digestibility as well as the silages without additives. The major changes on the chemical composition were noticed after the day 7 of fermentation. For the addition of urea and the combination of microorganisms L. buchneri and Pediococcus the variation was better described by linear equations whereas quadratic and cubic effects were more suitable for fitting the data from the control and the L. buchneri added silages. Even tough all silages has shown high acetic acid contents, mainly the combination of lactic bacteria, no significant effects were observed upon the yeast counts. However, the levels of lactic acid and propionic acid and the ratio of both over the acetic acid content were related to the decrease on the activity of the alcohol dehydrogenase enzyme and, furthermore, on the ethanol content of the silages. The cluster analysis based on molecular evaluation demonstrated a change promoted over the bacterial population mediated by the additives applied during the ensiling of sugarcane.

京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第22148号 / 医博第4539号 / 新制||医||1039(附属図書館) / 京都大学大学院医学研究科医学専攻 / (主査)教授 坂井 義治, 教授 小西 靖彦, 教授 滝田 順子 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

Molecular biological methods are fast and efficient tool for the identification of microorganisms in real samples. The aim of this diploma thesis was analysis of microflora in control and contaminated brined cheeses. DNA isolated from analyzed samples was used to optimize the PCR course using primers with GC clamp on the distribution of amplicons using DGGE. DGGE products were reamplified after optimization and prepared for DNA sequencing. DNA isolated from analyzed samples was used in real-time PCR with high resolution melt analysis of the amplicons (HRMA). Samples of cheese and bacterial cultures isolated from cheeses were compared by DGGE and HRMA. Comparing the position of the amplicons was found that contaminants may be Bacillus licheniformis, Staphylococcus epidermidis, and Acinetobacter baumanii/ calcoaceticus. Sequence analysis of cheese and pickles amplicons, the presence of Bacillus sp. and other microorganisms spree in five genera were detected. Representatives of the tree genera were cultured. It is considered contamination Bacillus sp., or microorganisms which are not culturable methods used. The method is suitable for the analysis of complex microflora in cheese and pickles after further optimization.

In all organisms are rising a reactive oxygen and nitrogen species by the effects of various stress factors and these species have a negative impact on the organism. Due to this species plants have built up an efficient antioxidant system, that helps them to resist negative effects of reactive oxygen and nitrogen species. In this work was researched the effect of hydrogen peroxide and sodium benzoate on the production of hydrogen peroxide, superoxide, reactive nitrogen species and malondialdehyde, contained in the root and above-ground part of maize (Zea mays L.). By use of the fluorescence microscopy there were obtained images of cross-cut of root from which was determined the intensity of fluorescence of individual parts of the root and was examined the effect of the intensity of fluorescence markers of oxidative stress in dependence on the type of the fluorescence filter used.

This diploma thesis deals with the study of possible interactions between hyaluronan (HA) and plasmid DNA (pDNA). Plasmid DNA was isolated from E. coli JM109 (pUC19) and resuspended in TE buffer as well as high molecular weight hyaluronan. Individual samples of pDNA, HA and pDNA-HA were characterized by gel electrophoresis, CD spectroscopy and high resolution ultrasonic spectroscopy. Agarose gel electrophoresis examined the effect of the addition of hyaluronan to plasmid DNA on the migration of samples to the positive electrode. Structural changes in pDNA-HA samples were examined using CD spectroscopy. Individual CD spectra describes the dependence of the difference in absorption coefficients for left-hand and right-handed elliptic polarized light at wavelength. High resolution ultrasonic spectroscopy has been used to study interactions. It is an analytical method based on ultrasonic velocity and attenuation. We classify this technique as a non-destructive method because the passing waves do not affect the structure of the analyzed sample.

Tato diplomová práce se zabývá vlivem zaplňovacích činidel na interakce v systému DNA-tenzid. DNA o velikosti 4017 párů bází byla připravena polymerázovou řetězovou reakcí, jako templát byl použit plasmid pSB-E1g. Polyetylen glykol (PEG) byl použit jako zaplňovací činidlo a jeho vliv na DNA-tenzid interakce byl zkoumán experimenty založenými na fluorescenci a gelové elektroforéze. Také byl studován vliv iontové síly za použití NaBr na interakce DNA-tenzid za použití zaplňovacího činidla. Data byla vyhodnocena a evaluována v této práci. V úvahu byl brán i možný vliv polyetylen glykolu na kritickou micelarní koncentraci (CMC) tenzidu, bylo provedeno měření CMC pomocí ultrazvuku s vysokým rozlišením, avšak nebyl zjištěn žádný značný vliv zaplňovacího činidla na CMC tenzidu. Část této práce bude zahrnuta v publikaci s anglickým názvem Combined role of macromolecular crowding and cationic surfactant in efficient DNA condensation.

Stomata are small pores that are located on the surface of epidermal leaves, and they can regulate the uptake of CO2 and prevent water lose by opening and closing the pores. Stomata of plants can be regulated by external condition such as CO2, biotic and abiotic stresses and internal factors. CAM (crassulacean acid metabolism) plants adapt to hot and dry environments by closing stomata during the day and opening stomata during the cool night. However, it is still unclear how CAM plants open their stomata during the night and close them during the day. In this study, a number of factors were evaluated for their potential roles in promoting stomatal opening in the model CAM plant Kalanchoe laxiflora. Citrate is an important organic acid and it accumulates during the night in CAM plants. It is shown in this study that citrate promoted stomatal opening in detached leaf epidermis of Kalanchoe laxiflora. Further, the cytokinin zeatin is also shown to stimulate stomatal opening in detached leave of Kalanchoe laxiflora. Melatonin is an important regulator of circadian rhythms in mammals and has been implicated in regulation of plant abiotic stress responses. Melatonin was detected in the leaves of Kalanchoe laxiflora. It promoted stomatal opening in detached epidermis of Kalanchoe laxiflora. Together, these results suggest that stomata of Kalanchoe laxiflora respond to citrate and malate which are the main organic acids accumulate during nighttime and also to some signaling molecules (zeatin, melatonin, and serotonin) by opening stomata during dark period.

Rice (Oryza sativa L.) is an important crop cultivated worldwide, and abiotic stresses limit its productivity. Different approaches were carried out to understand the mechanisms of rice defense responses against abiotic stresses, mainly drought. The NADPH-generating enzymes engaged in response to dehydration and salt stresses during the seedling stage of Nipponbare cultivar were analyzed. Enzyme activities of 6-phosphogluconate dehydrogenase (6PGDH), NADP-dependent aldehyde dehydrogenase, NADP-malic enzyme (NADP-ME) in leaves, and 6PGDH in roots were significantly increased in response to dehydration stress. NADP-ME and NADP-glutamate dehydrogenase activities in roots increased significantly in response to salt stress. These results suggest the involvement of NADPH-generating enzymes in plant responses to dehydration and salinity stresses, and the increased demands of NADPH in plants under abiotic stress can be furnished by enhanced activities of NADPH-producing enzymes. Also, a dehydration-induced protein was detected and identified as serine-hydroxymethyltransferase. This result indicates that serine-hydroxymethyltransferase can play a key role in regulating dehydration response in rice. Moreover, comparative proteomic analyses of CL163 (drought-tolerant), Cheniere (drought-sensitive), and Rex (moderately-drought-sensitive) rice varieties were performed. Drought-responsive proteome changes were profiled in leaves and roots at the seedling stage in response to drought stress imposed by polyethylene glycol (PEG-6000). Eighteen significantly differentially expressed proteins were identified by mass spectrometry. Elongate factor1 alpha and 17.9-kDa classI heat shock protein appear to have different expression patterns between CL163 and Cheniere, which may be attributable to the difference in drought response of the two rice varieties. Furthermore, a compendium of 103 drought resistance genes in rice was compiled to construct and analyze networks formed by associations between genes/proteins and to identify the most significant genes, biological processes/pathways. Genes were classified based on gene ontology and protein class into 26 groups. Forty-two genes were classified as transcription factors. Proteins encoded by the genes were localized in 8 subcellular locations and classified into three classes. Two pathways from KEGG whose genes were overrepresented in the compendium were identified. Gene expression, network presenting pairwise interactions between genes/proteins, and co-expression network were constructed. This study provides a systematic view of the crucial genes that can be contributing collectively to drought tolerance.