Comparative Transcriptome Analysis Revealed Candidate Genes Potentially Related to Desiccation Sensitivity of Recalcitrant Quercus variabilis Seeds

Chinese cork oak (Quercus variabilis) is a widely distributed and highly valuable deciduous broadleaf tree from both ecological and economic perspectives. Seeds of this species are recalcitrant, i.e., sensitive to desiccation, which affects their storage and long-term preservation of germplasm. However, little is known about the underlying molecular mechanism of desiccation sensitivity of Q. variabilis seeds. In this study, the seeds were desiccated with silica gel for certain days as different treatments from 0 (Control) to 15 days (T15) with a gradient of 1 day. According to the seed germination percentage, four key stages (Control, T2, T4, and T11) were found. Then the transcriptomic profiles of these four stages were compared. A total of 4,405, 4,441, and 5,907 differentially expressed genes (DEGs) were identified in T2 vs. Control, T4 vs. Control, and T11 vs. Control, respectively. Among them, 2,219 DEGs were overlapped in the three comparison groups. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis showed that these DEGs were enriched into 124 pathways, such as “Plant hormone signal transduction” and “Glycerophospholipid metabolism”. DEGs related to hormone biosynthesis and signal transduction (ZEP, YUC, PYR, ABI5, ERF1B, etc.), stress response proteins (LEA D-29, HSP70, etc.), and phospholipase D (PLD1) were detected during desiccation. These genes and their interactions may determine the desiccation sensitivity of seeds. In addition, group specific DEGs were also identified in T2 vs. Control (PP2C62, UNE12, etc.), T4 vs. Control (WRKY1-like, WAK10, etc.), and T11 vs. Control (IBH1, bZIP44, etc.), respectively. Finally, a possible work model was proposed to show the molecular regulation mechanism of desiccation sensitivity in Q. variabilis seeds. This is the first report on the molecular regulation mechanism of desiccation sensitivity of Q. variabilis seeds using RNA-Seq. The findings could make a great contribution to seed storage and long-term conservation of recalcitrant seeds in the future.

Assessing the storage potential of Australian rainforest seeds: a decision-making key to aid rapid conservation

Seed banking of rainforest species is hindered by lack of knowledge as to which species are tolerant of desiccation and freezing. We assessed 313 Australian rainforest species for seed banking suitability by comparing the germination percentage of fresh seeds to seeds dried at 15% RH and seeds stored at −20 °C after drying. We then compared desiccation responses to environmental, habit, fruit and seed characteristics to identify the most useful predictors of desiccation sensitivity. Of 162 species with ≥ 50% initial germination, 22% were sensitive to desiccation, 64% were tolerant and 10% were partially tolerant; the responses of 4% were uncertain. Of 107 desiccation tolerant species tested for response to freezing, 24% were freezing sensitive or short-lived in storage at −20 °C. Median values for fresh seed moisture content (SMC), oven dry weight (DW) and the likelihood of desiccation sensitivity (PD-S) were significantly greater for desiccation sensitive than desiccation tolerant seeds. Ninety-four to 97% of seeds with SMC < 29%, DW < 20 mg or PD-S < 0.01 were desiccation tolerant. Ordinal logistic regression of desiccation response against environmental, habit, fruit and seed characteristics indicated that the likelihood of desiccation sensitivity was significantly increased by a tree habit, fleshy fruit, increasing fresh SMC and increasing PD-S. The responses observed in this study were combined with earlier studies to develop a simple decision key to aid prediction of desiccation responses in untested rainforest species.

Desiccation tolerance and sensitivity of selected tropical montane species in Sri Lanka

Although the level of seed desiccation sensitivity (LSDS) may have an impact on plant species conservation, information is available for <10% of tropical angiosperms. A study was conducted to assess the LSDS of 28 tropical montane species in Sri Lanka. Seeds were extracted from freshly collected fruits. Initial weight was recorded, and thousand seed weight (TSW) was calculated. Seed moisture content (MC) was determined. LSDS was determined using seed desiccation experiments and predicted using the TSW–MC criterion. Seed storage behaviour was predicted using LSDS and storage data and using a model based on phylogenetic affiliation. The relationship between LSDS and seed dormancy, life form and forest strata was evaluated. Fresh seeds of only 12 species germinated to >80%. Although seeds of the other species had >80% viability, only 0–70% germinated due to dormancy. Seeds of five species had MC <15%, indicating desiccation tolerance (DT). Seeds of 12 species lost viability after desiccation, indicating desiccation sensitivity (DS). Seeds of Ardisia missionis, Psychotria gartneri and Psychotria nigra remained viable after desiccation, showing DT. Seeds of 17 species were DS and those of 11 species DT. The TSW of four species was >500 g. Thus, seeds of other species were predicted to be DT by the TSW–MC criterion. A relationship was identified between LSDS and the forest strata of the species. More canopy species produced DS than DT seeds. Since seeds of most of the studied species were DS, these species may be threatened due to prolonged droughts predicted for the region due to climate change.

Comparative Transcriptome Analysis Revealed Candidate Genes Potentially Related to Desiccation Sensitivity of Recalcitrant Quercus variabilis Seeds

Background: Chinese cork oak (Quercus variabilis) is a widely distributed and highly valuable deciduous broadleaf tree from both ecological and economic perspectives. Seeds of Q. variabilis are recalcitrant, i.e., sensitive to desiccation, which affects their storage and long-term preservation of germplasm. However, little is known about the underlying mechanism of desiccation sensitivity of Q. variabilis seeds. Results: In this study, the seeds were desiccated with silica gel for 0 day (control, CK), one day (T1) to 15 days (T15). After desiccation, the transcriptomic profiles of these different desiccation stages were compared using the Quercus suber genome as a reference, as well four key stages (CK, T2, T4 and T11) of desiccation sensitivity of Q. variabilis seeds through germination test were identified. A total of 4405, 4441, and 5907 differentially expressed genes (DEGs) were identified in T2 vs CK, T4 vs CK, and T11 vs CK, respectively. Among them, 2219 DEGs were overlapped in the three comparison groups. KEGG (Kyoto Encyclopaedia of Genes and Genomes) enrichment analysis showed that these DEGs were enriched into 124 pathways, such as "plant hormone signal transduction" and "glycerophospholipid metabolism". DEGs related to hormone synthesis and signal transduction (ZEP, YUC, PYR, ABI5, ERF1B, etc), stress response proteins (LEA D-29, HSP70, etc), and phospholipase D (PLD1) were detected during seed desiccation. These genes and their interactions may regulate the desiccation sensitivity of Q. variabilis seeds. Finally, a possible work model was proposed to show the molecular regulation mechanism of desiccation sensitivity in recalcitrant Q. variabilis seedsConclusions: Our study is the first on the molecular regulation mechanism of desiccation sensitivity of Q. variabilis seeds by using RNA-Seq and propose a possible work model. Our findings could make a great contribute to seed storage and long-term conservation of germplasm resources of recalcitrant seeds in the future.

Variation in seed traits among Mediterranean oaks in Tunisia and their ecological significance

Background and Aims Oaks are the foundation and dominant tree species of most Mediterranean forests. As climate models predict dramatic changes in the Mediterranean basin, a better understanding of the ecophysiology of seed persistence and germination in oaks could help define their regeneration niches. Tunisian oaks occupy distinct geographical areas, which differ in their rainfall and temperature regimes, and are thus a valuable model to investigate relationships between seed traits and species ecological requirements. Methods Seed morphological traits, desiccation sensitivity level, lethal freezing temperature, embryonic axis and cotyledon sugar and lipid composition, and seed and acorn germination rates at various constant temperatures were measured in Quercus canariensis, Q. coccifera, Q. ilex and Q. suber, using seeds sampled in 22 Tunisian woodlands. Key Results Only faint differences were observed for desiccation sensitivity in the oak species studied. By contrast, the species differed significantly in sensitivity to freezing, germination rates at low temperature and base temperature. Quercus ilex and Q. canariensis, which occur at high elevations where frost events are frequent, showed the lowest freezing sensitivity. A significant correlation was found between hexose contents in the embryonic axis and freezing tolerance. Significant interspecific differences in the time for seeds to germinate and the time for the radicle to pierce the pericarp were observed. The ratio of pericarp mass to acorn mass differed significantly among the species and was negatively correlated with the acorn germination rate. Quercus coccifera, which is frequent in warm and arid environments, showed the highest acorn germination rate and synchrony. Conclusions Seed lethal temperature, seed germination time at low temperatures, the ratio of pericarp mass to acorn mass and the embryonic axis hexose content appeared to be key functional traits that may influence the geographical ranges and ecological requirements of Mediterranean oaks in Tunisia.