The origin of the agriculture was one of the turning points in human history, and a central part of this was the evolution of new plant forms, domesticated crops. Seed dispersal and germination are two key traits which have been selected to facilitate cultivation and harvesting of crops. The objective of this study was to analyze anatomical structure of seed coat and pod, identify metabolic compounds associated with water-impermeable seed coat and differentially expressed genes involved in pea seed dormancy and pod dehiscence. Comparative anatomical, metabolomics, and transcriptomic analyses were carried out on wild dormant, dehiscent Pisum elatius (JI64, VIR320) and cultivated, indehiscent Pisum sativum non-dormant (JI92, Cameor) and recombinant inbred lines (RILs). Considerable differences were found in texture of testa surface, length of macrosclereids, and seed coat thickness. Histochemical and biochemical analyses indicated genotype related variation in composition and heterogeneity of seed coat cell walls within macrosclereids. Liquid chromatography–electrospray ionization/mass spectrometry and Laser desorption/ionization–mass spectrometry of separated seed coats revealed significantly higher contents of proanthocyanidins (dimer and trimer of gallocatechin), quercetin, and myricetin rhamnosides and hydroxylated fatty acids in dormant compared to non-dormant genotypes. Bulk Segregant Analysis coupled to high throughput RNA sequencing resulted in identification of 770 and 148 differentially expressed genes between dormant and non-dormant seeds or dehiscent and indehiscent pods, respectively. The expression of 14 selected dormancy-related genes was studied by qRT-PCR. Of these, expression pattern of four genes: porin (MACE-S082), peroxisomal membrane PEX14-like protein (MACE-S108), 4-coumarate CoA ligase (MACE-S131), and UDP-glucosyl transferase (MACE-S139) was in agreement in all four genotypes with Massive analysis of cDNA Ends (MACE) data. In case of pod dehiscence, the analysis of two candidate genes (SHATTERING and SHATTERPROOF) and three out of 20 MACE identified genes (MACE-P004, MACE-P013, MACE-P015) showed down-expression in dorsal and ventral pod suture of indehiscent genotypes. Moreover, MACE-P015, the homolog of peptidoglycan-binding domain or proline-rich extensin-like protein mapped correctly to predicted Dpo1 locus on PsLGIII. This integrated analysis of the seed coat in wild and cultivated pea provides new insight as well as raises new questions associated with domestication and seed dormancy and pod dehiscence.
We use a relatively densely sampled phylogeny to obtain preliminary estimates for the ages of the major clades identified in Euphorbia, with which we show that the succulent species of Euphorbia have diversified over the last 36 million years into many of the semi–arid, tropical parts of the world. Many major clades have subclades from widely separated regions, often on different continents. Our results imply that these distributions arose by long–distance dispersal after the break–up of Gondwana. In the case of Indian/South–east Asian succulents there appears to have been a single, relatively recent dispersal event from Africa. We have included many species from the Arabian Peninsula and Socotra and we show that these are nested within other, mainly African clades. In some cases Arabian and Socotran taxa have their closest relatives in adjacent parts of North–east Africa and most often this is true in recent clades, while in more ancient clades their closest relatives may be in Macaronesia or in the Namib Desert of Southern Africa so that these are typical 'Rand Flora' elements. In one case, closest known relatives are in North America. We find that Socotran taxa vary between 16 and 3 Ma old. The major diversifications of succulents in temperate Southern Africa (the crown clades in subg. Rhizanthium and more minor clades in subg. Chamaesyce) and in tropical East Africa (the crown clades in subg. Euphorbia) occurred in the last 20–3 Ma. In the Greater Cape Flora of the western part of Southern Africa the diversity in Euphorbia is mainly derived from one lineage in subg. Rhizanthium and one in subg.Chamaesyce. In contrast the diversity of Euphorbia in the Arabian Peninsula is derived from the invasion (mainly from Africa) of many separate lineages. We show that large, succulent trees are only found in subg. Euphorbia and only occur in the Old World. Most of them fall within a single clade and have evolved relatively recently.
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