Sidney Netto Parentoni

Mapping QTLs for aluminum tolerance in maize

Aluminum toxicity is one of the major constraints for plant development in acid soils, limiting food production in many countries. Cultivars genetically adapted to acid soils may offer an environmental compatible solution, providing a sustainable agriculture system. The aim of this work was to identify genomic regions associated with Al tolerance in maize, and to quantify the genetic effects on the phenotypic variation. A population of 168F3:4 families derived from a cross between two contrasting maize inbred lines for Al tolerance was evaluated using the NSRL and RSRL parameters in nutrient solution containing toxic level of aluminum. Variance analyses indicated that the NSRL was the most reliable phenotypicindex to measure Al tolerance in the population, being used for further QTL mapping analysis. RFLP and SSR markers were selected for bulked segregant analysis, and additional SSR markers, flanking the polymorphisms of interest, were chosen in order to saturate the putative target regions. Seven linkage groups were constructed using 17 RFLP and 34 SSR markers. Five QTLs were mapped on chromosomes 2, 6 and 8, explaining 60% of the phenotypic variation. QTL4 and marker umc043 were located on chromosomes 8and 5, close to genes encoding for enzymes involved in the organic acids synthesis pathways, a widely proposed mechanism for Al tolerance in plants. QTL2 was mapped in the same region as Alm2,also associated with Al tolerance in maize. In addition, dominant and additive effects were important in the control of this trait in maize.

Hematoxylin staining as a phenotypic index for aluminum tolerance selection in tropical maize (Zea mays L.)

Abstract Hematoxylin staining is an early indicator of Aluminum (Al) toxicity effects on the apices of young, developing roots grown in nutrient solution. In this work, the potential of this technique as a reliable and reproducible phenotypic index for Al tolerance in tropical maize genotypes was assessed, with its performance systematically compared to two other parameters widely used in breeding programs – relative seminal-root length (RSRL) and net seminal-root length (NSRL). Seeding roots from contrasting genotypes for Al sensitivity stained remarkably different after 24- and 48-h and 7-day exposures to 222 μM Al in nutrient solution, with the Al-dye complex being detected in both the outer (epidermis) and inner (cortex) portions of the roots from the sensitive cultivar. Hematoxylin staining was compared to the RSRL and NSRL parameters using 20 families from the third generation of selfing (S3) following the cross between two contrasting inbred lines that had been previously classified by the RSRL index in an independent procedure. The coloration technique showed the highest capacity to discriminate among tolerant and sensitive genotypes and displayed significant correlation coefficients to the other two indexes. Evaluation of the results from diallel crosses involving nine inbred lines proved that hematoxylin staining was also particularly adequate for identifying expressive hybrid vigor, as demonstrated by the general (GCA) and specific (SCA) combining ability estimates obtained by using the three indexes simultaneously. Hence, hematoxylin staining of Al-stressed root apices appears to be a powerful tool to assist in Al-tolerance selection in tropical maize breeding programs.

A role for root morphology and related candidate genes in P acquisition efficiency in maize

Phosphorus (P) is an essential nutrient for plants and is acquired from the rhizosphere solution as inorganic phosphate. P is one of the least available mineral nutrients, particularly in highly weathered, tropical soils, and can substantially limit plant growth. The aim of this work was to study a possible effect of root morphology and the expression pattern of related candidate genes on P efficiency in maize. Our field phenotyping results under low and high P conditions enabled us to identify two contrasting genotypes for P acquisition efficiency that were used for the root traits studies. Root morphology was assessed in a paper pouch system to investigate root traits that could be involved in P acquisition efficiency. The genes, Rtcs, Bk2 and Rth3, which are known to be involved in root morphology, showed higher expression in the P efficient line relative to the P inefficient line. Overall, root traits showed high heritability and a low coefficient of variation. Principal component analysis revealed that out of the 24 root traits analysed, only four root traits were needed to adequately represent the diversity among genotypes. The information generated by this study will be useful for establishing early selection strategies for P efficiency in maize, which are needed to support subsequent molecular and physiological studies.

Aluminum Effects on Nitrogen Uptake and Nitrogen Assimilating Enzymes in Maize Genotypes with Contrasting Tolerance to Aluminum Toxicity

Abstract The objective of this study was to investigate the effect of aluminum (Al) on NO3 − and NH4 + uptake and on activity/expression of the N‐assimilating enzymes glutamine synthetase (GS1, GS2, and GSr isoforms, EC 6.3.1.2), glutamate synthase (NADH‐GOGAT, EC 1.4.7.14; ferredoxin‐GOGAT, EC 1.4.7.1), and glutamate dehydrogenase (GDH, EC 1.4.1.2) of maize (Zea mays L.) genotypes contrasting in tolerance to Al toxicity. Seeds were germinated on paper towels, screened for uniformity and allowed to grow for 5–6 d in a modified Steinberg nutrient solution, pH 5.5, in a glasshouse, under natural light. Immediately after transference to a growth chamber (12 h photoperiod, 26°C ± 0.5/18°C ± 0.9 day/night temperature, 72% RH and 540 µ Em−2 s−1 quantum flux density) Al stress was imposed by lowering pH to 4.5 and addition of 166 µmoles L−1 AlCl3. Control plants were grown in the same nutrient solution without Al, pH 4.5. Aluminum inhibited NO3 − and NH4 + uptake and decreased shoot dry matter yield in all genotypes used in this investigation. Irrespective of their tolerance to Al toxicity, Al‐treated plants had less nitrogen (N), phosphorus (P), calcium (Ca), magnesium (Mg), and sulfur (S) in their leaves. In roots, Al treatment did not affect dry matter weight but decreased accumulation of N, Ca, and Mg. Phosphorus and potassium (K) tended to accumulate in roots of Al‐treated plants, suggesting that Al inhibited P translocation to the leaves. In roots, Al induced anaplerotic GDH and phosphoenolpyruvate carboxylase (PEPC, EC 4.1.1.31) activities, but inhibited GS activity. Moreover, in the apex (0–5 mm) and in the 6–15 mm segment of the elongation zone of most genotypes, Al tended to inhibit expression of GSr and favor expression of GS1. This increase in GS1 expression concomitantly with decreased N uptake and elevated deaminating GDH activity are indicative of metabolic changes usually associated with plant senescence. In leaves, Al did not influence GOGAT and GS activities and expression of GS isoforms suggesting it does not disturb assimilation of nitrogenous compounds translocated from roots to leaves. Nonetheless, lower leaf PEPC activity suggests that Al intoxicated plants had lower CO2 − fixation rates. The inverse effect of Al treatment on the activities of GDH and PEPC in roots and leaves, indicates that whereas increased activities in roots were induced by stress related stimuli, decreased activities in leaves were caused by a reduced nitrogen supply to the leaves. Because the earliest effects observed in this investigation appeared only after 24 h of exposure to Al and were similar in both tolerant and sensitive genotypes, it is concluded that the changes observed in N uptake and assimilation into plant amino acids were downstream of the initial, primary Al toxicity event.

Adaptability and stability of maize varieties using mixed model methodology

The objective of this study was to evaluate the performance, adaptability and stability of corn cultivars simultaneously in unbalanced experiments, using the method of harmonic means of the relative performance of genetic values. The grain yield of 45 cultivars, including hybrids and varieties, was evaluated in 49 environments in two growing seasons. In the 2007/2008 growing season, 36 cultivars were evaluated and in 2008/2009 25 cultivars, of which 16 were used in both seasons. Statistical analyses were performed based on mixed models, considering genotypes as random and replications within environments as fixed factors. The experimental precision in the combined analyses was high (accuracy estimates > 92 %). Despite the existence of genotype x environment interac- tion, hybrids and varieties with high adaptability and stability were identified. Results showed that the method of harmonic means of the relative performance of genetic values is a suitable method for maize breeding programs.

Inbreeding depression of 28 maize elite open pollinated varieties

The study of inbreeding depression is important for breeding strategies such as use of inbred progenies or extraction of inbreed lines. A diallel of 28 maize open-pollinated varieties was evaluated in 10 environments in the early 1990s. At the same time, S1 populations for each of the 28 varieties were evaluated in the same 10 experiments (environments). Yield reductions of the populations from S0 to S1 (mean of the 10 environments), varied from 34.6% (CMS-01) to 59.2% (CMS-30), with an average of 49.1%. Inbreeding depression was greater in populations with a wider genetic base, which had never been exposed to inbreeding (CMS-30, BR-107, PH4, Cunha, Saracura, Nitrodent, and Nitroflint). Inbred lines with greater yield means should be obtained from the BR-105, BR-111, CMS-01, CMS-03, BR-106, CMS-14c, and CMS-28 populations. The use of parameter estimates generated by analysis of inbreeding depression, allow to make inferences about frequencies of deleterious alleles in the population. The frequencies of favorable alleles in the parents can be obtained by diallel analysis. The association of these two types of information, can provide a better interpretation of the genetic parameters and also can improve the process of selection of parents for either an intra- or an inter-populational breeding program.

Genetic dissection of Al tolerance QTLs in the maize genome by high density SNP scan

BackgroundAluminum (Al) toxicity is an important limitation to food security in tropical and subtropical regions. High Al saturation on acid soils limits root development, reducing water and nutrient uptake. In addition to naturally occurring acid soils, agricultural practices may decrease soil pH, leading to yield losses due to Al toxicity. Elucidating the genetic and molecular mechanisms underlying maize Al tolerance is expected to accelerate the development of Al-tolerant cultivars.ResultsFive genomic regions were significantly associated with Al tolerance, using 54,455 SNP markers in a recombinant inbred line population derived from Cateto Al237. Candidate genes co-localized with Al tolerance QTLs were further investigated. Near-isogenic lines (NILs) developed for ZmMATE2 were as Al-sensitive as the recurrent line, indicating that this candidate gene was not responsible for the Al tolerance QTL on chromosome 5, qALT5. However, ZmNrat1, a maize homolog to OsNrat1, which encodes an Al3+ specific transporter previously implicated in rice Al tolerance, was mapped at ~40 Mbp from qALT5. We demonstrate for the first time that ZmNrat1 is preferentially expressed in maize root tips and is up-regulated by Al, similarly to OsNrat1 in rice, suggesting a role of this gene in maize Al tolerance. The strongest-effect QTL was mapped on chromosome 6 (qALT6), within a 0.5 Mbp region where three copies of the Al tolerance gene, ZmMATE1, were found in tandem configuration. qALT6 was shown to increase Al tolerance in maize; the qALT6-NILs carrying three copies of ZmMATE1 exhibited a two-fold increase in Al tolerance, and higher expression of ZmMATE1 compared to the Al sensitive recurrent parent. Interestingly, a new source of Al tolerance via ZmMATE1 was identified in a Brazilian elite line that showed high expression of ZmMATE1 but carries a single copy of ZmMATE1.ConclusionsHigh ZmMATE1 expression, controlled either by three copies of the target gene or by an unknown molecular mechanism, is responsible for Al tolerance mediated by qALT6. As Al tolerant alleles at qALT6 are rare in maize, marker-assisted introgression of this QTL is an important strategy to improve maize adaptation to acid soils worldwide.

Eficiência de índices fenotípicos de comprimento de raiz seminal na avaliação de plantas individuais de milho quanto à tolerância ao alumínio

The objective of this work was to verify the efficiency of the relative seminal root length (RSRL) and net seminal root length (NSRL) as phenotypic indexes for aluminum tolerance in individual maize plants. Seedlings of Al tolerant and susceptible genotypes were grown in nutrient solution containing toxic level of Al for a period of seven days, after which the values of RSRL and NSRL were determined. The results obtained when mean values of RSRL and NSRL were utilized showed that both indexes were able to discriminate tolerant from susceptible maize genotypes. However, when individual values of RSRL and NSRL were utilized, some tolerant plants scored as susceptible ones were observed, showing that the phenotypic evaluation of individual plants by both indexes is subjected to significant errors, mainly in the characterization of susceptible plants. Therefore, in studies for quantitative traits loci (QTLs) mapping for Al tolerance in maize, where these indexes are used, the most appropriate is to use F 3 families, where it is possible to obtain mean values for RSRL and NSRL and then utilize these means to represent the phenotypic values of the respective F 2 plants.

AVANÇOS NO PROCESSO SELETIVO DA VARIEDADE DE MILHO PIPOCA BRS ANGELA

A populacao e a distribuicao espacial de plantas na cultura do milho sao importantes para a interceptacao da radiacao, a qual e fundamental para o rendimento. A interceptacao da radiacao e funcao da latitude do local, do dia do ano, da hora do dia e do ângulo foliar. Nao se dispoe de metodologia para previsao da melhor populacao e distribuicao espacial de plantas em funcao da arquitetura da copa, sendo ambos, em geral, determinados por experimentacao em campo. Tambem nao se dispoe de metodologia para previsao do ângulo foliar que maximize a interceptacao da radiacao. Os objetivos do trabalho foram propor equacoes e metodologias para previsao da populacao e da distribuicao espacial de plantas em funcao da arquitetura da copa, bem como de ângulos foliares que maximizem a interceptacao da radiacao em funcao do local e do dia do ano. A metodologia proposta deve ser validada pela comparacao dos resultados de rendimento obtidos de genotipos com diferentes arquiteturas de copa e cultivados em diferentes latitudes e periodos do ano.O objetivo deste trabalho foi avaliar o efeito de genotipos de sorgo sobre a biologia do pulgao Rhopalosiphum maidis (Fitch). Os experimentos foram conduzidos na Embrapa Milho e Sorgo, em Sete Lagoas, MG. Foram testados oito genotipos de sorgo, sendo sete de reconhecida resistencia ao pulgao Schizaphis graminum (Rondani), e a cultivar comercial BRS 303. Para se obter folhas para a criacao dos pulgoes em laboratorio, foi plantada, semanalmente, no campo, uma fileira de cinco metros lineares de cada material. Os experimentos foram conduzidos durante duas geracoes do pulgao, utilizando-se, na primeira, insetos recem-nascidos, com aproximadamente uma hora de idade e oriundos da cultivar BR 304. As ninfas foram individualizadas e mantidas em secoes foliares provenientes de cada uma das oito cultivares estudadas. Como segunda geracao, utilizaram-se ninfas recem-nascidas, obtidas de progenitores criados em cada genotipo estudado na fase anterior, ou seja, individuos da segunda geracao. Em ambas as geracoes, os insetos foram mantidos em câmaras climatizadas reguladas a 25 ± 1oC, UR 70 ± 10% e fotofase de 12 horas, avaliando-se o numero de instares, periodos prereprodutivo, reprodutivo e pos-reprodutivo, ciclo total, numero de ninfas produzidas por femea e peso dos insetos aos quatro e oito dias de idade, com 15 repeticoes. O delineamento experimental foi o inteiramente casualizado, em esquema fatorial (genotipos x geracoes x repeticoes). Os genotipos TX 430 (IS x SC) e GR 1.1.1.1.1 apresentaram efeito adverso sobre a biologia do inseto, reduzindo a sobrevivencia, a capacidade reprodutiva, o periodo reprodutivo, pos-reprodutivo e o ciclo total. O genotipo BRS 303 foi o hospedeiro mais adequado, proporcionando uma elevada producao de ninfas, alta viabilidade e periodos reprodutivo, pos-reprodutivo e ciclo de vida mais longos.As doencas do milho causadas por molicutes (fitoplasma e espiroplasma) destacam-se em importância para a cultura, em consequencia da alta incidencia e dos prejuizos que causam a producao de graos. Os molicutes infectam o floema das plantas; sendo assim, podem alterar a sintese de acucares e proteinas. O objetivo deste trabalho foi verificar o efeito dos molicutes na sintese de acucares soluveis totais e de proteinas em duas cultivares de milho. O experimento foi conduzido em vasos de plastico contendo 30kg solo previamente analisado e adubado, com duas plantas por vaso. O delineamento experimental utilizado foi um fatorial 3 x 2 inteiramente casualizado, com seis repeticoes; sendo tres inoculacoes (fitoplasma, espiroplasma e sadia) e duas cultivares (Dina 766 e BR 201). Trinta dias apos a inoculacao, foram realizadas medidas de temperatura foliar, umidade relativa, resistencia estomatica e transpiracao. No estadio de grao leitoso, uma das plantas foi colhida e seccionada em tres partes, para analise de acucares soluveis totais, nutrientes e materia seca. Foi realizada uma amostragem do internodio abaixo da primeira espiga e da folha bandeira, para determinacao da atividade da peroxidase e do conteudo proteico. A planta restante foi conduzida ate o final do ciclo, quando foram avaliados: numero de espigas, peso de espigas e de graos e materia seca dos graos. A analise de variância de todas as caracteristicas avaliadas nao detectou significância na interacao cultivar x inoculacao. Alguns parâmetros mostraram significância para cultivares, para inoculacao ou para ambos. Acucares soluveis, materia seca, atividade da peroxidase e conteudo proteico foram semelhantes em ambas as cultivares e nas inoculacoes. A resistencia estomatica foi maior para Dina 766 e nas plantas inoculadas por fitoplasma. A proliferacao de espigas foi maior nas plantas inoculadas pelos molicutes que nas sadias. O peso de espigas e de graos foi maior nas plantas sadias e com fitoplasma que naquelas infectadas por espiroplasma. Concentracao de nutrientes e quantidade absorvida, em geral, foram semelhantes, excetuando P e Zn. Os molicutes nao interferiram na concentracao dos acucares soluveis e no conteudo proteico total; entretanto, os dados de produtividade sugerem uma maior susceptibilidade das cultivares ao espiroplasma.

Evaluation of maize core collection for drought tolerance

The maize genebank (GBmaize) preserves nearly 4,000 accessions for conservation and use. The use is however restricted because the accessions do not perform as well as the elite genotypes. This problem can be reduced by pre-breeding, i.e., by extending the information on germplasm and introgressing useful alleles. Since irregular rainfall distribution and drought induce maize yield losses, drought tolerance is a main breeding target. In this study, the GBmaize accessions were evaluated for drought tolerance. Environmental factors, genotypes and the respective interactions influence the phenotypic expression. There was however no interaction genotype - irrigation level, so no accessions with different performance under the two water regimes could be identified. The performance of the following accessions was promising for a number of traits: SP154, BA166, MG099, CE002, SE025, BA154, BA194, BA085, MG076, PR053, Roxo Macapa, SE016, and AL018.