Abstract
American ginseng (Panax quinquefolius L.) is an important herb that is cultivated in China, North American, and South Korea. It is propagated from seed, but the seed has deep dormancy characteristics described as morphophysiological dormancy. Two-stage temperature stratification, a warm (15–20°C) and cold (2°C) stratification period of 6 months, has been used successfully for seed dormancy release. However, little is known about the molecular mechanisms of seed dormancy release in the stratification process. In this study, seed development after pollination and seed development in the dormancy release process were investigated in American ginseng. The transcriptome during seed dormancy release was analyzed using RNA-Seq technology and 78,207 unigenes (mean length 531 bp) were generated. Based on similarity searches of public databases, 54,292 of the unigenes (69.4%) were functionally annotated. Further, three digital gene expression (DGE) libraries were sequenced and differences in gene expression at three stages during seed cold stratification were examined. The greatest number of differentially expressed genes occurred in the 90DCS versus 180DCS libraries, while the lowest number of differentially expressed genes occurred in the 135DCS verus 180DCS libraries. GO enrichment analysis revealed that 59, 29, and 39 GO terms were significantly enriched in the biological process, molecular function, and cell component GO categories, respectively. There were 25,190 genes with KEGG pathway annotation in the three DGE libraries and their enrichment pathways were compared. The gene expressions of 30 selected unigenes were validated using quantitative PCR. This study is the first to provide the transcriptome sequences for seed dormancy release in American ginseng, and demonstrates the successful use of DGE profiling data for analyzing transcriptomic variation during dormancy release. These data provide a basis for future researches of seed dormancy in morphophysiological dormancy seeds in non-model plants.
Highlights
American ginseng (Panax quinquefolius L.), one of the most widely used medicinal plants of the Araliaceae family, has been introduced and cultivated in China for over forty years
The brief development stage of an American ginseng seed after pollination is shown in Fig. 1 (a–j)
We evaluated the digital gene expression (DGE) library using 30 unigenes including those that were associated with gibberellic acid (GA) and abscisic acid (ABA), as well as DELLA, aminocyclopropane-1-carboxylate synthase (ACC) (1-aminocyclopropane-1carboxylate synthase), GIGATEA, PICKLE (CHD3-type chromatin-remodeling factor PICKLE), KAO (Ent-kaurenoic acid oxidase), and CTR(serine/threonine-protein kinase CTR1) and others [36,37,38]
Summary
American ginseng (Panax quinquefolius L.), one of the most widely used medicinal plants of the Araliaceae family, has been introduced and cultivated in China for over forty years. The embryo in the newly harvested seed is not fully developed (average length 0.5mm), and needs a warm (15–20°C) and cold (2°C) stratification period of 6 to 18 months [1, 2, 3]. The other method is a two-stage process: a warm stratification stage during which the embryo begins to grow, and a cold stage during which the embryo completes its development and the endogenous dormancy of the seed is overcome. The two-stage ginseng seed stratification technology has been widely used [1, 6, 7], the internal molecular mechanism of seed dormancy release is still unclear
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