Ontogeny of the pollinium in Hoya carnosa provides new insights into microsporogenesis.

The presence of a pollinium is a distinct character in Apocynaceae which is important for phylogenetic analysis. The pollinium of Hoya has an outer sporopollenin wall and a pellucid margin which are adaptive features. However, their ontogeny and related evolutionary implications are not entirely understood. Therefore, a representative species Hoya carnosa was selected to investigate the pollinium development using light and electron microscopy and cytochemical tests. In contrast to the microsporogenesis in most angiosperms, which is associated with callose, the non-callosic intersporal walls in Hoya carnosa, together with the successive cytokinesis and linear form of the tetrad, represent an alternative pattern of microsporogenesis. This pattern has specific implication for the early stages of pollen morphogenesis. The absence of exine and apertures in the pollen grains in the pollinium could result from a combination of factors including the absence of callose in the early stages and the modifications in later developmental pathways, e.g., the sporopollenin accumulation pathway. The pollinium wall is an exine without stratification, its surface lacks sculptures, and it provides structural support and protection. The pollen tubes germinate through the pellucid margin and germinating ridge which are specialized features. The pellucid margin originates from aborted microspores. The germinating ridge that lies on the outer side of the pellucid margin develops in the same way as a classic pollen exine. The pollen grains are aggregated by intine fusion which is favorable for tube germination and growth. Comparing Asclepiadoideae with the other two subfamilies of Apocynaceae that develop a pollinium, the pollinium of Asclepiadoideae has reduced deposition of sporopollenin in the inner walls but an increase in the outer pollinium wall, thus making the inner walls more reduced and simplified, and the outer walls more solid. The adaptive characters of the pollen wall structure and the cohesion mechanism suggest that the pollinium of Hoya carnosa is a derived form of pollen aggregation.

Diverse function of the PISTILLATA, APETALA 3, and AGAMOUS-like MADS-box genes involved in the floral development in Alpinia hainanensis (Zingiberaceae).

Zingiberaceae is the vital clue and key node in the decreased process of fertile stamens in Zingiberales, helping to understand the evolution of the ginger families. This study focuses on Alpinia hainanensis to investigate the function of B- and C-class MADS-box genes in floral development. The introns size of two B-class genes AhPI and AhAP3, and one C-class gene AhAG are quite variable. By contrast, the positions of the corresponding introns are conserved, resulting in a similar exon size in homologs. The typical region 70 bp-CCAATCA element was not found in the second intron of AhAG compared to AG homologs. The subcellular localization showed that AhAP3 was in both intranuclear and extranuclear. The heterodimer was formed between APETALA3 and PISTILLATA but not between the B- and C-class proteins using Y2H and BiFC. The 35S::AhAG heterologous transformed Arabidopsis had curly and smaller rosette leaves with early flowering. Floral organs had no homeotic conversion, albeit sepals and petals reduced in size. Siliques development was affected and displayed wrinkled and shorter. By contrast, 35S::AhAP3 and 35S::AhPI did not show any modified phenotype in transgenic Arabidopsis thaliana. We first proposed the model for Alpinia flower development. MADS-box transcription factor binding at particular genomic locations and interaction with partners may be crucial for the development of the floral organ.

Corrigendum: Transcriptome Analysis of Hamelia patens (Rubiaceae) Anthers Reveals Candidate Genes for Tapetum and Pollen Wall Development.

[This corrects the article on p. 1991 in vol. 7, PMID: 28119704.].

Transcriptome Analysis of Hamelia patens (Rubiaceae) Anthers Reveals Candidate Genes for Tapetum and Pollen Wall Development.

Studies of the anther transcriptome on non-model plants without a known genome are surprisingly scarce. RNA-Seq and digital gene expression (DGE) profiling provides a comprehensive approach to identify candidate genes contributing to developmental processes in non-model species. Here we built a transcriptome library of developing anthers of Hamelia patens and analyzed DGE profiles from each stage to identify genes that regulate tapetum and pollen development. In total 7,720 putative differentially expressed genes across four anther stages were identified. The number of putative stage-specific genes was: 776 at microspore mother cell stage, 807 at tetrad stage, 322 at uninucleate microspore stage, and the highest number (1,864) at bicellular pollen stage. GO enrichment analysis revealed 243 differentially expressed and 108 stage-specific genes that are potentially related to tapetum development, sporopollenin synthesis, and pollen wall. The number of expressed genes, their function and expression profiles were all significantly correlated with anther developmental processes. Overall comparisons of anther and pollen transcriptomes with those of rice and Arabidopsis together with the expression profiles of homologs of known anther-expressed genes, revealed conserved patterns and also divergence. The divergence may reflect taxon-specific differences in gene expression, the use RNA-seq as a more sensitive methodology, variation in tissue composition and sampling strategies. Given the lack of genomic sequence, this study succeeded in assigning putative identity to a significant proportion of anther-expressed genes and genes relevant to tapetum and pollen development in H. patens. The anther transcriptome revealed a molecular distinction between developmental stages, serving as a resource to unravel the functions of genes involved in anther development in H. patens and informing the analysis of other members of the Rubiaceae.

Molecular basis of floral petaloidy: insights from androecia of Canna indica.

Floral organs that take on the characteristics of petals can occur in all whorls of the monocot order Zingiberales. In Canna indica, the most ornamental or 'petaloid' parts of the flowers are of androecial origin and are considered staminodes. However, the precise nature of these petaloid organs is yet to be determined. In order to gain a better understanding of the genetic basis of androecial identity, a molecular investigation of B- and C-class genes was carried out. Two MADS-box genes GLOBOSA (GLO) and AGAMOUS (AG) were isolated from young inflorescences of C. indica by 3' rapid amplification of cDNA ends polymerase chain reaction (3'-RACE PCR). Sequence characterization and phylogenetic analyses show that CiGLO and CiAG belong to the B- and C-class MADS-box gene family, respectively. CiAG is expressed in petaloid staminodes, the labellum, the fertile stamen and carpels. CiGLO is expressed in petals, petaloid staminodes, the labellum, the fertile stamen and carpels. Expression patterns in mature tissues of CiGLO and CiAG suggest that petaloid staminodes and the labellum are of androecial identity, in agreement with their position within the flower and with described Arabidopsis thaliana expression patterns. Although B- and C-class genes are important components of androecial determination, their expression patterns are not sufficient to explain the distinct morphology observed in staminodes and the fertile stamen in C. indica.