Molecular Control of Sporophyte-Gametophyte Ontogeny and Transition in Plants

Alternation of generations between a sporophytic and gametophytic developmental stage is a feature common to all land plants. This review will discuss the evolutionary origins of these two developmental programs from unicellular eukaryotic progenitors establishing the ability to switch between haploid and diploid states. We will compare the various genetic factors that regulate this switch and highlight the mechanisms which are involved in maintaining the separation of sporophytic and gametophytic developmental programs. While haploid and diploid stages were morphologically similar at early evolutionary stages, largely different gametophyte and sporophyte developments prevail in land plants and finally allowed the development of pollen as the male gametes with specialized structures providing desiccation tolerance and allowing long-distance dispersal. Moreover, plant gametes can be reprogrammed to execute the sporophytic development prior to the formation of the diploid stage achieved with the fusion of gametes and thus initially maintain the haploid stage. Upon diploidization, doubled haploids can be generated which accelerate modern plant breeding as homozygous plants are obtained within one generation. Thus, knowledge of the major signaling pathways governing this dual ontogeny in land plants is not only required for basic research but also for biotechnological applications to develop novel breeding methods accelerating trait development.

Initiation of microcalli in culture of pea (Pisum sativum L.) isolated microspores

Innovative biotechnologies based on use of double haploids enables developing new varieties considerably faster compared to conventional plant breeding approaches. In pea, reliable methods of haploid plants production are not fully elaborated yet. The current research aimed at testing different conditions (genotype, medium, and stress treatment) for initiation of sporophytic developmental shift in culture of pea isolated microspores. Reprogramming pea microspores towards a sporophytic development was stimulated with temperature stress. Cold (+4°С) and heat (+35°С) stress treatments were applied to pea isolated flower buds and microspores, respectively. Microspores were isolated from plants of 6 pea genotypes, treated at 18 temperature regimes and cultivated in 8 liquid nutrient media with various chemical compounds including growth regulators, vitamins, sugars, glutamine, casein hydrolysate, and osmotic agents. Microcalli were produced from isolated microspores of pea breeding line 109b and variety Stabil in conditions of nutrient media KM-ар1 and MSB-M3 after cold (+4°С) stress treatment for 16 and 10 days, respectively. The media KM-ар1 and MSB-M3 contained a relatively low concentration of sugar (10 and 6 g L-1, respectively), and were supplemented with polyethylene glycol 6000 or mannitol as osmotic agents.

Germination and sporophytic development of Regnellidium diphyllum Lindman (Marsileaceae) in the presence of hexavalent chromium

Regnellidium diphyllum Lindman is a heterosporous fern, growing in aquatic environments and surrounding wetlands, which is assumed to be threatened by increasing water pollution and disappearance of its natural habitats. Among contaminants, hexavalent chromium - Cr(VI) - is known to be present in effluents from some leather tanning factories. Megaspore germination tests were performed using Meyer's solution, at concentrations 0 (control), 0.1, 0.5, 1, 5, 10, 15, 20, 30, 50, and 80 mg.L-1, from a standard solution of Titrisol® 1000 mg.L-1. The primary development of apomictic sporophytes was studied using solutions containing 0.025 to 4.8 mg.L-1 of Cr(VI). The experiments were conducted in a growth chamber at 24 ± 1 ºC and for a 12-hour photoperiod under fluorescent lights, providing a nominal irradiance of 77 µmol.m-2/s. Significant differences in megaspore germination, with subsequent sporophytic development, were verified from 0.5 mg.L-1 Cr(VI) concentration onwards. Growth of primary root and primary and secondary leaves was significantly reduced at 3.2 mg.L-1 Cr(VI) concentration or higher. Considering the pollution from Cr(VI) in some areas of R. diphyllum natural occurrence, these data indicate that low reproductive rates and disappearance of populations are likely to occur in these situations.

Effects of 2,4-D on the germination of megaspores and initial development of Regnellidium diphyllum Lindman (Monilophyta, Marsileaceae)

Regnellidium diphyllum is considered as endangered, occurring in the State of Rio Grande do Sul, Brazil, and a few adjoining localities in Uruguay, Argentina and the State of Santa Catarina. It grows in wetlands frequently altered for agricultural activities. Herbicides based on 2,4-dichlorophenoxyacetic acid (2,4-D) are widely used in these fields. The effects of 2,4-D on the germination of megaspores and initial sporophytic development of R. diphyllum were investigated. Six concentrations of 2,4-D (0.32; 0.64; 1.92; 4.80; 9.60 and 19.20 mg.L-1), and the control (0.00 mg.L-1), were tested in vitro, using Meyer's medium. Cultures were maintained in a growth chamber at 24 ± 1 °C, under artificial light with nominal irradiance of 110 µmol.m-2/s and 16 hours photoperiod. Megaspore germination was lower at 9.60 and 19.20 mg.L-1 of 2,4-D (56 and 48%, respectively), compared with the control (68%). Herbicide concentrations of up to 1.92 mg.L-1 did not significantly decrease the number of sporophytes formed. At 19.20 mg.L-1, no sporophytes were formed. The lengths of the primary root, primary and secondary leaves were greater at concentrations of 0.32 and 0.64 mg.L-1 of 2,4-D. Low concentrations of 2,4-D do not affect germination rates and initial development of R. diphyllum in a significant way. However, higher concentrations (9.60 and 19.20 mg.L-1) affect substantially the germination of the megaspores and interfere with the establishment of the species.