Resumo: A germinação da semente do café (Coffea arabica L.) é resultado de eventos que ocorrem simultaneamente no embrião e endosperma. Para entender os mecanismos moleculares responsáveis por estes eventos, nós realizamos uma análise transcricional do embrião e dos endospermas micropilar e lateral de sementes embebidas por 10 dias. O sequenciamento obteve contigs que codificam 16.813 proteínas. Destas, 14.005 (~ 83%) foram altamente similares a pelo menos uma sequência de proteína na base de dados nr. 162 genes foram significativamente expressos em embrião, 36 em endosperma micropilar e 72 em endosperma lateral. A análise de tecido-especificidade dos genes significativamente expressos mostrou que a maior proporção de genes específicos está no embrião (113/162, ~70%), enquanto 11 genes foram expressos nos endospermas lateral e micropilar. No embrião, os genes foram associados principalmente com estresse abiótico, crescimento celular e comunicação intercelular. Nos endospermas micropilar e lateral, eles foram associados com estresse abiótico e degradação da parede celular. A acurácia da análise de RNA-seq foi confirmada por RT-qPCR. Este trabalho acrescenta novas informações sobre os mecanismos moleculares envolvidos na germinação de sementes de café.

Abstract: Germination of the coffee (Coffea arabica L.) seed is the result of events that occur simultaneously in the embryo and endosperm. To understand the molecular mechanisms responsible for these events, we undertook a transcriptome analysis of embryo, micropylar and lateral endosperms from 10-day-imbibed seeds. The sequencing yielded contigs coding for 16,813 proteins. From those, 14,005 (~ 83%) were highly similar to at least one protein sequence in the nr database. 162 genes were significantly expressed in the embryo, 36 in the micropylar endosperm and 72 in the lateral endosperm. The tissue specificity analysis of the significantly expressed genes showed that the embryo had the highest proportion of specific genes (113/162, ~70%), while 11 were expressed in the micropylar and lateral endosperms. In the embryo, genes were mainly associated with abiotic stress, cell growth, and intercellular communication. In the micropylar and lateral endosperms, they were associated with abiotic stress and cell wall degradation. The accuracy of RNA-seq data was confirmed by RT-qPCR. This work adds new information about the molecular mechanism involved in coffee seed germination.

Resumo: A sobrevivência durante o armazenamento ou longevidade é pré-requisito para evitar a deterioração, que leva à perda de vigor. A longevidade é rotineiramente avaliada pela habilidade da semente em germinar depois do armazenamento, que aumenta progressivamente durante a maturação depois da aquisição da tolerância à dessecação. No entanto, a germinação representa somente uma parte do processo de estabelecimento da planta. Foi avaliado como a maturação afeta a resistência de várias características, como vigor, relacionadas ao estabelecimento das plântulas em sementes de soja da variedade BRS 284. Três novos estádios entre 7.1 e 7.2 (7.1.1, 7.1.2 e 7.1.3) foram introduzidos para capturar o rápido aumento na longevidade das sementes. O armazenamento induziu uma queda na velocidade de germinação a partir de 7-14 dias de armazenamento, dependendo do estádio, e essa queda diminuiu com a maturação, a partir do estádio 7.1.3. O tempo para perda de 50% na capacidade de alongamento de ambos os órgãos foi similar àquele observado para capacidade de germinação. Além disso, esse tempo aumentou de forma constante durante o período de maturação, demonstrando a importância das fases finais de maturação da semente na construção do vigor.

Abstract: Long-term survival during dry storage or longevity is a pre-requisite to avoid deterioration, leading to loss of vigor. Longevity is routinely evaluated by the ability to germinate after storage. It increases progressively during seed maturation, after the acquisition of desiccation tolerance. However, the capacity to germinate represents only a part of the success of crop establishment. How seed maturation affects the resistance of several traits, as vigor, associated with seedling establishment, against deterioration was evaluated during seed filling and post-abscission phase of soybean BRS 284 seeds. Three new phenological stages between 7.1 and 7.2 (7.1.1, 7.1.2 and 7.1.3) were introduced to capture the rapid increase in seed longevity. Germination speed started to be affected at 7-14 days after storage depending on the stages. The delay on germination increased with maturation from 7.1.3 to dry mature seeds. The time to 50% loss of elongation capacity of both organs during storage was similar to that of loss of germination. Also, it increased steadily during seed maturation after mass maturity and harvest maturity stages, highlighting the importance of the late phase of seed maturation for building seed vigor.

The genus Arachis (Fabaceae) native to South America, contains 80 species divided into nine sections, three of which contain species of special economic importance such as the cultivated peanut (Arachis hypogaea), belonging to the section Arachis, and some perennial forage species from sections Caulorrhizae and Rhizomatosae. We used microsatellite markers to assay genetic variability among 77 accessions of four species from section Rhizomatosae, the diploid Arachis burkartii (2n = 2x = 20) and the tetraploid Arachis glabrata, Arachis pseudovillosa and Arachis nitida (2n = 4x = 40). A total of 249 alleles were found in the fifteen loci analyzed and a high degree of intra and interspecific polymorphism was detected. The lowest intraspecific variation occurred in Arachis burkartii, while the smallest estimated interspecific value was between A. nitida and A. pseudovillosa and the largest was between A. burkartii and A. nitida. High observed heterozygosity was detected in A. glabrata. The diploid accessions grouped in one cluster and the tetraploid accessions in another. It was possible to distinguish all 77 accessions and the genetic distance between accessions could not be correlated with geographic origin.

The Arachis section is the most important of the nine sections of the genus Arachis because it includes the cultivated peanut, Arachis hypogaea. The genetic improvement of A. hypogaea using wild relatives is at an early stage of development in spite of their potential as sources of genes, including those for disease and pests resistance, that are not found in the A. hypogaea primary gene pool. Section Arachis species germplasm has been collected and maintained in gene banks and its use and effective conservation depends on our knowledge of the genetic variability contained in this material. Microsatellites are routinely used for the analysis of genetic variability because they are highly polymorphic and codominant. The objective of this study was to evaluate the transferability of microsatellite primers and the assay of genetic variability between and within the germplasm of some species of the Arachis section. Fourteen microsatellite loci developed for three different species of Arachis were analyzed and 11 (78%) were found to be polymorphic. All loci had transferability to all the species analyzed. The polymorphic loci were very informative, with expected heterozygosity per locus ranging from 0.70 to 0.94. In general, the germplasm analyzed showed wide genetic variation.

The genus Arachis currently comprises 69 described species, some of which have potential and real value as human and animal foods. These Arachis species have been collected and maintained in germplasm banks to provide material that can be used as sources of genes in breeding programs and for the selection of new cultivars. One of the principal objectives of germplasm conservation is the evaluation of genetic variability, which is best conducted using molecular markers. We investigated the use of heterologous primers to amplify microsatellite loci that could be used to evaluate genetic variability in Arachis germplasm. Fifteen microsatellite primer pairs were tested in 76 accessions of 34 species from the nine Arachis sections. The data indicated that heterologous primers were very useful in Arachis since they had high transferability among the species (91%) and allowed the amplification of very polymorphic putative loci, which allowed both the characterization of most accessions and to make inferences about the mating systems of some species analyzed. Our data also revealed that the germplasm analyzed showed high variability, even when represented by few accessions.