University of Sao Paulo, CENA, Av. Centenario 303, Piracicaba, SP, 13416-903, Brazil.
Brazilian Agricultural Research Corporation (Embrapa), Embrapa Genetic Resources and Biotechnology, Cx.Postal 02372, Brasilia, DF, 70.770-917, Brazil.
Protoplasma. 2023 Mar;260(2):571-587. doi: 10.1007/s00709-022-01802-w. Epub 2022 Aug 10.
Brachiaria, a genus from the Poaceae family, is largely cultivated as forage in Brazil. Among the most cultivated varieties of Brachiaria spp., B. brizantha cv. Marandu (syn. Urochloa brizantha) is of great agronomical importance due to the large areas cultivated with this species. This cultivar is apomictic and tetraploid. Sexual diploid genotype is available for this species. The difference in levels of ploidy among sexual and apomictic plants contributes to hindering Brachiaria breeding programs. The induction of haploids and double haploids is of great interest for the generation of new genotypes with potential use in intraspecific crosses. A key factor for the success of this technique is identifying adequate microspore developmental stages for efficient embryogenesis induction. Knowledge of the morphological changes during microsporogenesis and microgametogenesis and sporophytic tissues composing the anther is critical for identifying the stages in which microspores present a higher potential for embryogenic callus and somatic embryo through in vitro culture. In this work, morphological markers were associated with anther and pollen grain developmental stages, through histological analysis. Anther development was divided into 11 stages using morphological and cytological characteristics, from anther with archesporial cells to anther dehiscence. The morphological characteristics of each stage are presented. In addition, the response of stage 8 anthers to in vitro culture indicates microspores initiating somatic embryogenic pathway.
臂形草属(Brachiaria)是禾本科的一个属,在巴西被广泛用作饲料。在臂形草属(Brachiaria)的多种栽培品种中,Marandu 品种(又名 Urochloa brizantha)因其种植面积大而具有重要的农艺学意义。该品种是无融合生殖的四倍体。该物种存在有性二倍体基因型。有性和无融合生殖植物之间的倍性水平差异阻碍了臂形草属的育种计划。诱导单倍体和双单倍体对于产生具有种内杂交潜力的新基因型非常重要。该技术成功的一个关键因素是确定合适的小孢子发育阶段,以有效地诱导胚胎发生。了解小孢子发生和小配子发生过程中的形态变化以及构成花药的孢子体组织,对于确定小孢子在体外培养中具有更高的胚胎发生愈伤组织和体细胞胚胎形成潜力的阶段至关重要。在这项工作中,通过组织学分析,将形态学标记与花药和花粉粒发育阶段相关联。使用形态学和细胞学特征将花药发育分为 11 个阶段,从具原表皮细胞的花药到花药开裂。呈现了每个阶段的形态特征。此外,对 8 期花药的离体培养反应表明小孢子启动了体细胞胚胎发生途径。