Transcriptome sequences generated by next-generation sequencing (NGS) technologies can be utilized to rapidly detect and characterize a large number of gene-based microsatellites from different plants. Rhododendron rex Lévl. is a perennial woody species from the family Ericaceae and an endangered plant with high ornamental value endemic to Southwestern China. Nevertheless, the genetic and genomic information of R. rex remain unknown. In this study, we performed transcriptome sequencing for R. rex leaf samples, and generated large transcript sequences for functional characterization and development gene-associated SSR markers. A total of 164,242 unigenes were assembled and 115,089 (70.07%) unigenes were successfully annotated in public databases. In addition, a total of 15,314 potential EST-SSRs were identified, and the frequency of SSRs in the R. rex unigenes was 9.32%, with an average of one EST-SSR per 5.65 kb. The most abundant type was repeated di-nucleotide (54.63%), followed by mono- (26.03%) and tri-nucleotide (18.51%) repeats. Based on the SSR-containing sequence, 100 primer pairs were randomly selected and synthesized and used for assessment of the polymorphism. Thirty-six primer pairs were polymorphic and revealed polymorphism among 20 individuals from four R. rex populations. A total of 197 alleles were identified, with an average of 5.472 alleles per locus. The Polymorphism Information Content ranged from 0.154 to 0.870, with a mean of 0.482. The newly developed EST-SSR markers exhibited high transferability (58.33–83.33%) among the six subgenera. Thus, these novel EST-SSR markers developed would provide valuable sequence resources for population structure, genetic diversity analysis, and genetic resource assessments of R. rex and its related species.

In the present study, the distribution, population size and structure, habitat and stand disturbance of a critically endangered plant Euryodendron excelsum H. T. Chang were examined using field investigations. A total of 179 individual plants, including 23 adult trees, were found. The population was distributed in 10 highly isolated and fragmented patches. Half of the 10 patches had less than 10 individual plants and two patches had only one remnant adult; there were no adult plants in two of the patches. Although the overall population structure trends to a reversed 'J' shape, two fragile stages of E. excelsum limit its natural recruitment and regeneration: seed germination and seedling growth toward adulthood. The major threats for the survival of this species are its specific small population size and the high frequency of anthropogenic destruction. Based on the results, some strategies were put forward for the conservation of E. excelsum populations.

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Euryodendron excelsum H. T. Chang is a critically endangered plant from the familyTheaceae that is endemic to China. It is now present in only one remnant population in a very narrow range and with a highly isolated and fragmented distribution pattern. The species is distributed close to the local villages and faces extinction because of the high level of anthropogenic disturbance. Thus, conservation and restoration of this species is urgent. The species reproduces by seed, but its germination requirements and seedling emergence are rarely understood. In this study, the germination requirements, desiccation tolerance and seedling emergence of the species were studied under controlled laboratory conditions. The results indicated that seeds of E. excelsum were non-dormant. Optimal temperatures for germination were 15°C to 25°C; the germination percentage decreased and the mean germination time increased at high temperature. Seed germination was inhibited in the dark, suggesting that the seeds were positive photoblastic. Water stress also significantly inhibited germination percentage; no seeds germinated at 15% polyethylene glycol 6000. The fresh seeds had moderate moisture content of 28.6% and showed strong tolerance of dehydration. Thus, the seeds of E. excelsum tended to be orthodox. Seeds on the soil surface had the highest emergence percentage, which declined with depth of burial. Soil types significantly affected seedling emergence; seeds had higher emergence percentage in sandy soil than in mixed soil and clayed soil. Our study demonstrated that ex situ conservation and reintroduction of E. excelsum using seedling propagation from seeds is feasible.

Apterosperma oblata H. T. Chang is an endangered species in the Theaceae, and although it reproduces by seed its germination requirements are not known. To provide basic information for conservation and restoration programs, we studied the effects of temperature, light, desiccation, and storage on the germination of A. oblata seeds under laboratory conditions. Optimal temperatures for seed germination were 15 and 20°C. Photoperiod greatly affected final germination percentage, and constant darkness inhibited seed germination. Fresh harvested seeds had a high moisture content (56.4%), and they gradually lost their germination ability as dehydration occurred, suggesting that the seeds may be recalcitrant. Although short-term (2 months) dry storage at 4°C increased the germination percentage, the overall trend was a considerable decline in germination percentage with an increase in storage duration at 4°C and 20°C. Since seeds are recalcitrant and easy to germinate at maturity, we recommend seedlings establishment rather than seeds for conservation and restoration programs of the species.