ABSTRACT This study aimed to evaluate the efficiency of the parameters fresh leaf mass and stomatal density in the estimation of ploidy and pre-selection of putative banana autotetraploids. Young plants of the diploid cultivar Ouro (AA), previously subjected to in vitro polyploidization with the antimitotic amiprofos-methyl - APM (0, 10, 20, 30, 40, and 60 µM L-1) and colchicine (2.5 mM L-1) were evaluated for survival, height, number of leaves, pseudostem diameter, leaf disc fresh mass, and stomatal density. Ploidy was determined by flow cytometry in a random sample of 200 plants to find the relationship between the genomic content and the analyzed variables. Spearman’s correlation analysis showed a strong correlation (0.84**) between leaf disc fresh mass and tetraploid plants, as well as an absence of significant correlation between stomatal density and ploidy levels. Thus, the pre-selection of plants was carried out using a reference value of leaf disc fresh mass and resulted in 688 plants pre-selected as putative tetraploids. Among them, 318 were confirmed as tetraploids, 291 as diploids, and 79 as mixoploids after flow cytometry analysis. The pre-selection efficiency was 46.2% of tetraploid plants from the total pre-selected. The results indicate that the use of leaf disc fresh mass is a simple, practical, and promising method to estimate ploidy when a high number of plants is obtained in in vitro banana polyploidization studies, reducing the total number of plants to have confirmed ploidy.

RESUMO Este estudo objetivou avaliar a eficiência dos parâmetros massa fresca foliar e densidade estomática na estimativa da ploidia e pré-seleção de autotetraploides putativos de bananeira. Plantas jovens da cultivar diploide Ouro (AA), previamente submetidas à poliploidização in vitro com os antimitóticos amiprofos-metil - APM (0, 10, 20, 30, 40 e 60 µM L-1) e colchicina (2,5 mM L-1) foram avaliadas quanto à sobrevivência, altura, número de folhas, diâmetro do pseudocaule, massa fresca de disco foliar e densidade estomática. Determinou-se a ploidia, via citometria de fluxo, numa amostra aleatória de 200 plantas, a fim de encontrar relação entre o conteúdo genômico e as variáveis analisadas. Por meio da análise de correlação de Spearman, verificou-se forte correlação (0,84**) entre a massa fresca de disco foliar e as plantas tetraploides, bem como ausência de correlação significativa entre a densidade estomática e os níveis de ploidia. Assim, a pré-seleção das plantas foi feita mediante um valor de referência de massa fresca de disco foliar e resultou em 688 plantas pré-selecionadas como tetraploides putativos. Destas, 318 foram confirmadas como tetraploides, 291 como diploides e 79 mixoploides, após análise de citometria de fluxo. A eficiência de pré-seleção foi de 46,2% de plantas tetraploides do total pré-selecionado. Os resultados indicam que o uso da massa fresca de disco foliar é um método simples, prático e promissor para estimar a ploidia quando é obtido um elevado número de plantas em trabalhos de poliploidização in vitro da bananeira, reduzindo o total de plantas a ter a ploidia confirmada.

The aim of this work was to evaluate the carotenoid content and genetic variability of banana accessions from the Musa germplasm collection held at Embrapa Cassava and Tropical Fruits, Brazil. Forty-two samples were analyzed, including 21 diploids, 19 triploids and two tetraploids. The carotenoid content was analyzed spectrophotometrically and genetic variability was estimated using 653 DArT markers. The average carotenoid content was 4.73 µg.g-1, and ranged from 1.06 µg.g-1 for the triploid Nanica (Cavendish group) to 19.24 µg.g-1 for the triploid Saney. The diploids Modok Gier and NBA-14 and the triploid Saney had a carotenoid content that was, respectively, 7-fold, 6-fold and 9-fold greater than that of cultivars from the Cavendish group (2.19 µg.g-1). The mean similarity among the 42 accessions was 0.63 (range: 0.24 to 1.00). DArT analysis revealed extensive genetic variability in accessions from the Embrapa Musa germplasm bank.

The chromosomes of Crotalaria juncea, a legume of agronomic interest with a 2n = 16 karyotype composed of metacentric chromosomes, were analyzed using several cytogenetic techniques. C-banding revealed heterochromatic regions around the centromeres in all chromosomes and adjacent to the secondary constriction on the chromosome 1 short arm. Fluorescent staining with the GC-specific chromomycin A3 (CMA) highlighted these heterochromatic regions and a tiny site on the chromosome 1 long arm while the AT-specific stain 4'-6-diamidino-2-phenylindole (DAPI) induced a reversed pattern. Staining with CMA combined with AT-specific distamycin A (DA) counterstaining quenched the pericentromeric regions of all chromosomes, but enhanced fluorescence was observed at the heterochromatic regions around the secondary constriction and on the long arms of chromosomes 1 and 4. Fluorescence in situ hybridization (FISH) revealed 18S-5.8S-26S rRNA gene sites (45S rDNA) on chromosomes 1 and 4, and one 5S rDNA locus on chromosome 1. All the rDNA sites were co-located with the positive-CMA/DA bands, suggesting they were very rich in GC. Silver staining revealed signals at the main 45S rDNA locus on chromosome 1 and, in some cells, chromosome 4 was labeled. Two small nucleoli were detected in a few interphase cells, suggesting that the minor site on chromosome 4 could be active at some stages of the cell cycle.