Is a non-synonymous SNP in the HvAACT1 coding region associated with acidic soil tolerance in barley?

The barley HvAACT1 gene codes for a citrate transporter associated with tolerance to acidic soil. In this report, we describe a single nucleotide polymorphism (SNP) in the HvAACT1 coding region that was detected as T-1,198 (in genotypes with lower root growth on acidic soil) or G-1,198 (greater root growth) and resulted in a single amino acid change (L/V-172). Molecular dynamic analysis predicted that HvAACT1 proteins with L or V-172 were stable, although the substitution led to structural changes within the protein. To evaluate the effect of the SNP on tolerance to acidic soil, barley accessions were separated into haplotypes based on the presence of a 1 kb insertion in the HvAACT1 promoter and a 21 bp insertion/deletion. These markers and the SNP-1,198 allowed the identification of five haplotypes. Short-term soil experiments showed no difference in root growth for most of the accessions containing the 21 bp insertion and T or G-1,198. In contrast, genotypes showing both the 21 bp deletion and G-1,198, with one of them having the 1 kb insertion, showed greater root growth. These results indicate that the SNP was not advantageous or deleterious when genotypes from the same haplotype were compared. The occurrence of the SNP was highly correlated with the 21 bp insertion/deletion that, together with the 1 kb insertion, explained most of the barley tolerance to acidic soil.

Assessment of genetic diversity in Brazilian barley using SSR markers.

Barley is a major cereal grown widely and used in several food products, beverage production and animal fodder. Genetic diversity is a key component in breeding programs. We have analyzed the genetic diversity of barley accessions using microsatellite markers. The accessions were composed of wild and domesticated barley representing genotypes from six countries and three breeding programs in Brazil. A total of 280 alleles were detected, 36 unique to Brazilian barley. The marker Bmag120 showed the greatest polymorphism information content (PIC), with the highest mean value found on chromosome three, and the lowest on chromosomes four and six. The wild accessions presented the highest diversity followed by the foreign genotypes. Genetic analysis was performed using Principal Coordinates Analysis, UPGMA clustering, and Bayesian clustering analysis implemented in Structure. All results obtained by the different methods were similar. Loss of genetic diversity has occurred in Brazilian genotypes. The number of alleles detected in genotypes released in 1980s was higher, whereas most of the cultivars released thereafter showed lower PIC and clustered in separate subgroups from the older cultivars. The use of a more diverse panel of genotypes should be considered in order to exploit novel alleles in Brazilian barley breeding programs.

Improvement in Brazilian barley breeding: Changes in developmental phases and ecophysiological traits.

Despite recognizing the importance of genetic improvement in the production of barley grains, little information is available on the contribution of phenological development to the genetic improvement of Brazilian barley. Field experiments were carried out between 2011 to 2013, in the absence of biotic and abiotic stresses and with preventive lodging control. Five two-rowed spring barley cultivars, released between 1968 and 2008, were evaluated. Although there was no significant association in the cycle length (Emergence - Anthesis) of the cultivars with the year of release, the genetic improvement increased the proportion of the Doble ridge - Maximum number of spikelet primordia/Maximum number of spikelet primordia - anthesis period to the total time to anthesis. The period between DR-MNP was increased in modern cultivars, to the detriment of the Doble ridge - Maximum number of spikelet primordia period. However, the duration of the period between emergences to the double ridge (vegetative phase) was not altered in the analyzed period. Barley breeding in Brazil did not change the total number of leaves on the main stem but caused an increase in the number of leaves earlier in the development, favoring the high level of tillering. The leaf architecture of modern barley was altered towards a more vertical inclination (erectophilic canopy), allowing the penetration of photosynthetically active radiation into the crop canopy.

Characterization of the Fusarium sambucinum species complex and detection of multiple mycotoxins in Brazilian barley samples.

This study investigated the fungal diversity in Brazilian barley samples, focusing on the Fusarium sambucinum species complex and the presence of multiple mycotoxins: aflatoxins B1, B2, G1, G2 beauvericin (BEA), enniatins (ENNs) A, A1, B, and B1, deoxynivalenol (DON), fumonisins (FB) B1 and B2, HT-2 and T-2 toxins, nivalenol (NIV) and ochratoxin A (OTA) from two different regions, São Paulo (SP) and Rio Grande do Sul (RS). The majority of the isolates belonged to the Fusarium sambucinum species complex (FSAMSC), with F. graminearum s.s. characterized as the major contaminant. F. meridionale and F. poae were the second most frequent fungi isolated from SP and RS, respectively. All of the F. graminearum s.s. isolates demonstrated 15-ADON genotype, whereas F. poae and F. meridionale were all NIV. The majority of the F. cortaderiae isolates were NIV, with only one 3-ADON genotype. Mycotoxin analysis revealed that none of the samples were contaminated by aflatoxins, OTA, FB2 and type A trichothecenes, however, all of the samples were contaminated with at least one Fusarium toxin. Contamination by DON, ZEA, ENNB and ENNB1 levels were significantly higher in RS. Co-contamination of BEA, DON, ENNs, NIV and ZEA in 18.5% and 24.2% of the analyzed samples was observed, from SP and RS respectively. More than 20% of the samples from RS presented DON and ZEA levels above the regulations established by Europe and Brazil. The results provide further information on the FSAMSC from South America and detected multiple Fusarium toxins in barley samples. This highlights the importance for further studies on the possible interactions of these mycotoxins in order to determine potential risks to animal health.

Is a non-synonymous SNP in the HvAACT1 coding region associated with acidic soil tolerance in barley?

Abstract The barley HvAACT1 gene codes for a citrate transporter associated with tolerance to acidic soil. In this report, we describe a single nucleotide polymorphism (SNP) in the HvAACT1 coding region that was detected as T-1,198 (in genotypes with lower root growth on acidic soil) or G-1,198 (greater root growth) and resulted in a single amino acid change (L/V-172). Molecular dynamic analysis predicted that HvAACT1 proteins with L or V-172 were stable, although the substitution led to structural changes within the protein. To evaluate the effect of the SNP on tolerance to acidic soil, barley accessions were separated into haplotypes based on the presence of a 1 kb insertion in the HvAACT1 promoter and a 21 bp insertion/deletion. These markers and the SNP-1,198 allowed the identification of five haplotypes. Short-term soil experiments showed no difference in root growth for most of the accessions containing the 21 bp insertion and T or G-1,198. In contrast, genotypes showing both the 21 bp deletion and G-1,198, with one of them having the 1 kb insertion, showed greater root growth. These results indicate that the SNP was not advantageous or deleterious when genotypes from the same haplotype were compared. The occurrence of the SNP was highly correlated with the 21 bp insertion/deletion that, together with the 1 kb insertion, explained most of the barley tolerance to acidic soil.

Desenvolvimento de populações duplo-haplóides de cevada cervejeira associadas à atividade das enzimas (1-3, 1-4)-β-glucanases / Development of doubled-haploids populations in malting barley associated to activity of enzymes (1-3, 1-4)-β-glucanases

Populações duplo-haplóides apresentam especial vantagem para análises genéticas, uma vez que a informação que elas oferecem pode ser maximizada, devido ao fato que todos os locos encontram-se em homozigose. O objetivo deste trabalho foi o desenvolvimento de duas populações duplo-haplóides (DHs) de cevada (Hordeum vulgare ssp. vulgare L.) segregantes para a atividade das enzimas (1-3, 1-4)-β-glucanases, através da técnica de cultura de anteras. Foram realizados dois cruzamentos com cultivares contrastantes para esta característica. As cultivares parentais selecionadas foram 'MN 698' e 'CEV 97047', para o desenvolvimento da população "malte verde" (MV), e 'Embrapa 127' e 'CEV 96025' para o desenvolvimento da população "malte seco" (MS). Foram cultivadas 10.734 anteras da população MS e 4.139 anteras da população MV. A população MV produziu 50 por cento mais plântulas verdes quando comparada à população MS, refletindo a importância do genótipo na resposta à cultura de anteras e na regeneração. A maioria das plantas adultas duplo-haplóides foi obtida através da duplicação espontânea in vitro do genoma haplóide, ocorrendo em 66 por cento das plantas da população MS e 76 por cento das plantas da população MV. Também foram observadas, em menor frequência, plantas haplóides, triplóides e tetraplóides. Através da técnica de cultura de anteras, foram desenvolvidas 204 linhagens duplo-haplóides, das quais 72 linhagens são da população "malte seco" e 132 linhagens são da população "malte verde". Este material constitui um importante germoplasma para o melhoramento genético da cevada.

Doubled haploid populations offer special advantages in genetic analyses, since the information they provide may be maximized due to the fact that all loci are homozygous. The aim of this study was to develop two barley (Hordeum vulgare ssp.vulgare L.) doubled-haploid (DHs) populations segregating to (1-3, 1-4)-β-glucanases activity, utilizing the anther culture protocol. Two crosses were performed with contrasting cultivars to (1-3, 1-4)-β-glucanases activity. Parental cultivars used were 'MN 698' and 'CEV 97047' for the development of 'green malt' population (MV) and Embrapa 127 and 'CEV 96025' for the development of "dry malt" population (MS). For the MS and MV populations, 10,734 and 4,139 anthers were cultured, respectively. MV population produced 50 percent more green seedlings as compared to MS population, which reflects the importance of genotype to the culturing of anthers and to regeneration. Most doubled-haploid adult plants were obtained by in vitro spontaneous duplication of the haploid genome, which occurred in 66 percent of plants from the MS population and 76 percent of plants from the MV population. Haploid, triploid and tetraploid individuals were also observed, though at low frequencies. The anther culture protocol afforded to develop 204 double-haploid lineages, of which 72 were generated by the 'dry malt' population and 132 from the "green malt" population. This material should be considered as important germplasm for barley genetic improvement.

Genotypic differences in proembryoid development and green plantlets regeneration through androgenesis in barley varieties / Diferenças genotípicas no desenvolvimento de pró-embrióides e regeneração de plântulas verdes via androgênese em genótipo de cevada

The development of in vitro haploid plants followed by spontaneous or induced genome duplication allows to achieve, in one generation, the recovery of total homozygosis. The efficiency of the haplodiploidization process through in vitro anther culture of barley is variable among genotypes. This study was aimed at determining the androgenetic response of nine barley genotypes from the breeding program of Embrapa Trigo, analysing proembryoid development and green plantlets regeneration in anthers cultivated in vitro. Cultivar 'BR2' presented the highest average of proembryoids (104/anther) and 'MN698' presented the highest average of green plantlets (0,41/anther). There was a significant variation among the average values of barley genotypes for embryo formation and green plantlets regeneration, making possible the selection to combine androgenetic capacity and good agronomic traits.

A obtenção, na cevada, de plantas haplóides in vitro e a posterior duplicação natural ou artificial do genoma permitem alcançar a homozigose completa, em uma geração. A eficiência da haplodiploidização pela cultura de anteras é variável entre os genótipos. Foi avaliada a resposta androgenética através da formação de pró-embrióides e da regeneração de plântulas verdes em nove cultivares do programa de melhoramento de cevada da Embrapa Trigo, em anteras cultivadas in vitro. A cultivar "BR2" apresentou maior média de pró-embrióides (104/antera), enquanto "MN698" mostrou a maior média de plântulas verdes (0,41/antera). Houve variação significativa entre os valores médios dos genótipos em relação à formação de pró-embrióides e à regeneração de plântulas verdes, indicando a possibilidade de seleção para combinar a capacidade androgenética com boas características agronômicas.