Functional Identification of 9-cis-epoxycarotenoid Dioxygenase Genes in Double Dormant Plant-herbaceous Peony

Seed dormancy and germination is a complex process, which is affected by external environmental conditions and internal factors independently or mutually. Phytohormones play an important regulatory role in this process. ABA was the main phytohormone affecting herbaceous peony seed dormancy release. However, the mechanism of ABA in the dormancy release of herbaceous peony needs to be further explored. Here, transcriptome data was screened from the perspective of ABA metabolism, and significantly differentially expressed PlNCED1 and PlNCED2 were obtained. We found that their expression trends were positively correlated with ABA content. Among them, PlNCED2 had a stronger regulatory effect on ABA content and was more sensitive to exogenous ABA. Overexpression and silencing of PlNCEDs in callus could affect the expression of PlCYP707As and the content of endogenous ABA. Through the observation of seed germination of Arabidopsis thaliana (A. thaliana), we found PlNCED1 and PlNCED2 promoted seed dormancy, and the promotion effect of PlNCED2 was more obvious. In general, PlNCED1 and PlNCED2 participated in the dormancy release of herbaceous peony seeds by regulating the accumulation of endogenous ABA. Our work can reveal the molecular mechanism and related theories of ABA involved in herbaceous peony seed dormancy release.

Seed-coat thickness explains contrasting germination responses to smoke and heat by Leucadendron species

Fire stimulates germination of most seeds in fire-prone vegetation. Fruits of Leucadendron (Proteaceae) are winged achenes or nutlets that correlate with their requirements for smoke and/or heat in promoting germination. We describe five possible smoke–heat dormancy-release/germination syndromes among plants, of which Leucadendron displays three (no response, smoke only, smoke and heat). As seed-coat thickness varies with seed-storage location (plant or soil) and morphology (winged or wingless), we tested its possible role in water uptake and germination. Species with winged seeds achieved 100% germination in the absence of smoke/heat, seed coats were an order of magnitude thinner, and their permeability greatly exceeded that of nutlets. As seed-coat thickness increased a) imbibitional water uptake declined at a decreasing rate, and b) the response to smoke, and to a lesser extent heat, increased linearly to reach levels of germination approaching those of winged seeds. For species responsive to smoke and heat, there was no additive effect when applied together, suggesting that they may have promoted the same physiological process. By what mechanisms a) the smoke response is greater the thicker the seed coat, and b) smoke chemicals could increase water permeability to explain the non-additive effect of smoke and heat, warrant further investigation.

The coupled effect of light and temperature on dormancy release and germination of Pinus koraiensis seeds

Elucidating the regulatory mechanisms of environmental factors on seed dormancy and germination will provide guidance for tree regeneration. Toward understanding the coupled effect of light and temperature on dormancy release and germination of Pinus koraiensis seeds, we set up three light conditions (L200: 200 μmol m−2 s−1, L20: 20 μmol m−2 s−1, L0: 0 μ m−2 s−1) and four storage temperatures [T-5: − 5 °C (50 days), T5: − 5 °C (50 days) + 5 °C (50 days), T25: − 5 °C(50 days) + 5 °C (50 days) + 25 °C (50 days), T15: − 5 °C (50 days) + 5 °C (50 days) + 25 °C (50 days) + 15 °C (50 days)] using imbibed seeds, then quantified phytohormones gibberellic acid (GA3) and abscisic acid (ABA) during the stratification. Germination percentage (GP), mean germination time (TM), and germination value (GV) under 25/15 °C temperature and the three light conditions were then determined. Phytohormone levels and germination performances were significantly affected by light and temperature. No consistent trend was found between the phytohormone levels and GP caused by light levels. Under the three light conditions, ABA concentrations in the embryo and endosperm decreased as storage temperature shifted from T-5 to T25 and increased from T25 to T15; GA3 decreased in nearly all four storage temperatures. GP reached 40–60% in T25 storage without light irradiance. In the three light conditions, GP and GV were higher at T5 and T25 than at T-5 and T15; so T5 and T25 are considered as optimum storage temperatures for dormancy release and germination. At optimum temperatures, light (L200, L20) significantly increased the GP and GV compared with the dark (L0). At L200 and L20, significant negative correlations between GV and the ABA concentrations and positive correlations between GV and GA/ABA in the seed embryo were found. Temperature played a more important role in primary dormancy release and germination; light was unnecessary for primary dormancy release. Light facilitated seed germination at optimum temperatures. The dormancy release and germination of P. koraiensis seeds were controlled by a decrease in ABA concentrations or an increase in GA/ABA induced by temperature variations.

Cellular and Metabolite Changes in the Secondary Phloem of Chinese Fir (Cuninghamia lanceolata (Lamb.) Hook.) during Dormancy Release

Wood in the cold temperate zone is the product of the alternation of the growing season and the dormant period of trees, but our knowledge of the process of dormancy release in trees remains limited. Chinese fir (Cuninghamia lanceolata (Lamb.) Hook.) was used to investigate cellular and metabolite changes in the secondary phloem tissue during dormancy release. The sampling dates were 2 March, 28 March, and 13 April. The microsections of wood-forming tissue were prepared using the paraffin embedding technique to observe the formation of cambium cells; metabolites in secondary phloem cells were extracted using a methanol/chloroform organic solvent system. The results showed that the secondary phloem consists of phloem fibers, sieve cells and phloem parenchyma. The cells were regularly arranged in continuous tangential bands and were in the order of Phloem fiber-Sieve cell-Phloem parenchyma-Sieve cell-Phloem parenchyma-Sieve cell-Phloem parenchyma-Sieve cell-Sieve cell-Phloem parenchyma-. The Chinese fir cambium was in dormancy on 2 March and 28 March, while on 13 April, it was already in the active stage and two layers of xylem cells with several layers of phloem cells were newly formed. The width of the cambium zone increased from 18.7 ± 5.7 μm to 76.5 ± 3.0 μm and the average radial diameter of sieve cells expanded from 15.4 ± 7.5 μm to 21.5 ± 7.4 μm after dormancy release. The cambium zone width and the average radial diameter of sieve cells before and after dormancy release were significantly different (p < 0.01). The phloem parenchyma cells without resin were squeezed and deformed by the sieve cells, and the width of the phloem during the active period was 197.0 ± 8.5 μm, which was larger than that during the dormant period. Ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS)-based metabolomics was employed to analyze the secondary phloem of Chinese fir on 28 March and 13 April. Thirty-nine differential metabolites during dormancy release were detected. The results showed that the composition of Chinese fir metabolites was different before and after dormancy release. The relative increase in pyruvic acid and ascorbic acid contents proved that the rate of energy metabolism in Chinese fir increased substantially after dormancy release. Changes in cell development and the composition of metabolites revealed that the dormancy release of Chinese fir was at early April and the formation period of phloem tissue is earlier than xylem tissue.

Deep complex morphophysiological dormancy in seeds of Viburnum plicatum var. formosanum (Adoxaceae) from subtropical mountains

Viburnum is a temperate-zone genus that also occurs in mountains of South America and Malesia, and seeds of many species have morphophysiological dormancy (MPD). Information on the level of MPD in seeds of species in various clades of Viburnum potentially would increase our understanding of the evolutionary relationships between the nine levels of MPD. Our aim was to determine the level of MPD in seeds of Viburnum plicatum var. formosanum that is endemic to mountains (1800–3000 m a.s.l.) in Taiwan and a member of the Lutescentia clade. The temperature requirements for embryo growth and root and shoot emergence and response of seeds to gibberellic acid (GA) were determined. No fresh seeds germinated during 16 weeks of incubation at 15/5, 20/10, 25/15, 30/20 or 25°C. Embryo growth and root emergence occurred during moist cold stratification at 5°C or at a temperature sequence of 15/5 to 5°C. During cold stratification, embryos length increased from 0.76 ± 0.06 to 3.40 ± 0.26 mm and the embryo length:seed length ratio from 0.20 ± 0.02 to 0.68 ± 0.07. In a temperature sequence simulating field conditions, embryos grew inside seeds at 5°C, roots emerged at 15/5°C and shoots emerged at 20/10°C. The optimum temperature for embryo growth was 5°C. Neither GA3 nor GA4 was effective in promoting root emergence. We conclude that seeds of V. plicatum var. formosanum have deep complex MPD, which is a first report for Viburnum. Dormancy release during the cool season at high elevations helps to ensure that seeds germinate at the beginning of the warm season.

Transcriptome profiles revealed molecular mechanisms of alternating temperatures in breaking the epicotyl morphophysiological dormancy of Polygonatum sibiricum seeds

Background To adapt seasonal climate changes under natural environments, Polygonatum sibiricum seeds have a long period of epicotyl morphophysiological dormancy, which limits their wide-utilization in the large-scale plant progeny propagation. It has been proven that the controlled consecutive warm and cold temperature treatments can effectively break and shorten this seed dormancy status to promote its successful underdeveloped embryo growth, radicle emergence and shoot emergence. To uncover the molecular basis of seed dormancy release and seedling establishment, a SMRT full-length sequencing analysis and an Illumina sequencing-based comparison of P. sibiricum seed transcriptomes were combined to investigate transcriptional changes during warm and cold stratifications. Results A total of 87,251 unigenes, including 46,255 complete sequences, were obtained and 77,148 unigenes (88.42%) were annotated. Gene expression analyses at four stratification stages identified a total of 27,059 DEGs in six pairwise comparisons and revealed that more differentially expressed genes were altered at the Corm stage than at the other stages, especially Str_S and Eme. The expression of 475 hormone metabolism genes and 510 hormone signaling genes was modulated during P. sibiricum seed dormancy release and seedling emergence. One thousand eighteen transcription factors and five hundred nineteen transcription regulators were detected differentially expressed during stratification and germination especially at Corm and Str_S stages. Of 1246 seed dormancy/germination known DEGs, 378, 790, and 199 DEGs were associated with P. sibiricum MD release (Corm vs Seed), epicotyl dormancy release (Str_S vs Corm), and the seedling establishment after the MPD release (Eme vs Str_S). Conclusions A comparison with dormancy- and germination-related genes in Arabidopsis thaliana seeds revealed that genes related to multiple plant hormones, chromatin modifiers and remodelers, DNA methylation, mRNA degradation, endosperm weakening, and cell wall structures coordinately mediate P. sibiricum seed germination, epicotyl dormancy release, and seedling establishment. These results provided the first insights into molecular regulation of P. sibiricum seed epicotyl morphophysiological dormancy release and seedling emergence. They may form the foundation of future studies regarding gene interaction and the specific roles of individual tissues (endosperm, newly-formed corm) in P. sibiricum bulk seed dormancy.

Combining conventional QTL analysis and whole-exome capture-based bulk-segregant analysis provides new genetic insights into tuber sprout elongation and dormancy release in a diploid potato population

Tuber dormancy and sprouting are commercially important potato traits as long-term tuber storage is necessary to ensure year-round availability. Premature dormancy release and sprout growth in tubers during storage can result in a significant deterioration in product quality. In addition, the main chemical sprout suppressant chlorpropham has been withdrawn in Europe, necessitating alternative approaches for controlling sprouting. Breeding potato cultivars with longer dormancy and slower sprout growth is a desirable goal, although this must be tempered by the needs of the seed potato industry, where dormancy break and sprout vigour are required for rapid emergence. We have performed a detailed genetic analysis of tuber sprout growth using a diploid potato population derived from two highly heterozygous parents. A dual approach employing conventional QTL analysis allied to a combined bulk-segregant analysis (BSA) using a novel potato whole-exome capture (WEC) platform was evaluated. Tubers were assessed for sprout growth in storage at six time-points over two consecutive growing seasons. Genetic analysis revealed the presence of main QTL on five chromosomes, several of which were consistent across two growing seasons. In addition, phenotypic bulks displaying extreme sprout growth phenotypes were subjected to WEC sequencing for performing BSA. The combined BSA and WEC approach corroborated QTL locations and served to narrow the associated genomic regions, while also identifying new QTL for further investigation. Overall, our findings reveal a very complex genetic architecture for tuber sprouting and sprout growth, which has implications both for potato and other root, bulb and tuber crops where long-term storage is essential.