Commercial cultivars of garlic, a popular condiment, are sterile, making genetic studies and breeding of this plant challenging. However, recent fertility restoration has enabled advanced physiological and genetic research and hybridization in this important crop. Morphophysiological studies, combined with transcriptome and proteome analyses and quantitative PCR validation, enabled the identification of genes and specific processes involved in gametogenesis in fertile and male-sterile garlic genotypes. Both genotypes exhibit normal meiosis at early stages of anther development, but in the male-sterile plants, tapetal hypertrophy after microspore release leads to pollen degeneration. Transcriptome analysis and global gene-expression profiling showed that >16,000 genes are differentially expressed in the fertile vs. male-sterile developing flowers. Proteome analysis and quantitative comparison of 2D-gel protein maps revealed 36 significantly different protein spots, 9 of which were present only in the male-sterile genotype. Bioinformatic and quantitative PCR validation of 10 candidate genes exhibited significant expression differences between male-sterile and fertile flowers. A comparison of morphophysiological and molecular traits of fertile and male-sterile garlic flowers suggests that respiratory restrictions and/or non-regulated programmed cell death of the tapetum can lead to energy deficiency and consequent pollen abortion. Potential molecular markers for male fertility and sterility in garlic are proposed.

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Background: Geophytes possess specialized storage organs -bulbs, tubers, corms or rhizomes, which allow their survival during unfovarable periods and provide energy support for sprouting and sexual and vegetative reproduction. Bulbing and flowering of the geophyte depend on the combined effects of the internal and external factors, especially temperature and photoperiod. Many geophytes are extensively used in agriculture, but mechanisms of regulation of their flowering and bulbing are still unclear. Results: Comparative morpho-physiological and transcriptome analyses and quantitative validation of gene expression shed light on the molecular regulation of the responses to vernalization in garlic, a typical bulbous plant. Long dark cold exposure of bulbs is a major cue for flowering and bulbing, and its interactions with the genetic makeup of the individual plant dictate the phenotypic expression during growth stage. Photoperiod signal is not involved in the initial nuclear and metabolic processes, but might play role in the later stages of development, flower stem elongation and bulbing. Vernalization for 12 weeks at 4°C and planting in November resulted in flower initiation under short photoperiod in December-January, and early blooming and bulbing. In contrast, non-vernalized plants did not undergo meristem transition. Comparisons between vernalized and non-vernalized bulbs revealed~14,000 differentially expressed genes.Conclusions: Low temperatures stimulate a large cascades of molecular mechanisms in garlic, and a variety of flowering pathways operate together for the benefit of meristem transition, annual life cycle and viable reproduction results.The circadian clock appears to play a central role in the transition of the meristem from vegetative to reproductive stage in bulbous plant, serving as integrator of the low-temperature signals and the expression of the genes associated with vernalization, photoperiod and meristem transition. The reserved photoperiodic pathway is integrated at an upstream point, possibly by the same receptors. Therefore, in bulb, low temperatures stimulate cascades of developmental mechanisms, and several genetic flowering pathways intermix to achieve successful sexual and vegetative reproduction.

Juvenile and vegetative adult shoot apical meristems (SAM) are actively involved in acquisition of flowering competence, while the embryonic SAM is regarded as “responsible” only for germination. Comparative analyses of imbibed and stratified seeds of garlic Allium sativum show that only stratified seedlings produced bulbs and flower stems at the end of the season. Since the seed morphology of stratified and non-stratified seeds prior to sowing was similar, the differences are attributed to the molecular alterations in the embryonic SAM. Functional annotation analysis of 3000 differentially expressed genes suggests that seed imbibition reactivates the embryonic cell cycle, initiates the metabolism, and primes garlic seed germination. Stratification enhances DNA modifications, biosynthesis, cellular transport, and tissue development. Similar to vernalization of the vegetative buds, stratification of the embryonic SAM resulted in altered expression of meristem-identity and flowering homologs. Phase transitions from seed germination to flowering and bulbing in A. sativum are tightly connected, and possibly associated with downregulation of specific flowering repressor(s). The embryonic SAM plays an important role not only in seed germination, but in the entire plant life cycle, providing the foundation for the genetic regulation of major functional shifts in metabolism and development.

Lemon wormwood Artemisia balchanorum was recently introduced to southern Russia as a new aromatic plant. Based on biological and chemical characteristics, several populations with dominant citral, linalool, and geraniol production were selected for further development and maintained by seed propagation. Chemical analysis of five outstanding populations at three stages of annual development: vegetative, flower buds, and full flowering, confirmed that the seed populations retain the distinct dynamics of the dominant and minor components during the annual cycle and can be used for the commercial production of citral, linalool, and geraniol. Micropropagation in vitro allows for efficient clonal micropropagation and mass reproduction of elite cultivars and promising forms of A. balchanorum on a commercial scale but cannot serve as a source of direct and efficient production of secondary metabolites.