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The Cypripedium forrestii is an orchid species with extremely small populations (PSESP) in Yunnan, China. C. forrestii is range-restricted and less-studied than many orchid species, and it is exposed to various threats to its survival. We investigated its potential habitats and collected 52 samples from eight locations, as well as two outgroup species for reference. We developed genetic markers (SNPs) for C. forrestii based on transcriptome sequencing (RNA-seq) data, and analyzed the genetic diversity, population structure, gene flow and demographic history of C. forrestii in detail. C. forrestii is a taxonomically independent species to protect. We found that the genetic diversity of C. forrestii was very low (1.7e-4) compared with other endangered species. We identified three genetic clusters, and several populations with distinct genetic backgrounds. Most genetic diversity was found within sampling sites (87.87%) and genetic clusters (91.39%). Gene flow has been greatly limited over the most recent generations, probably due to geographical distance, historical climate change and habitat fragmentation. We also detected a severe bottleneck event brought about by the recent population constraints. These factors, together with its reproductive characteristics, contribute to the population fragmentation and low genetic diversity of C. forrestii. Based on our findings, we suggest an integrative conservation strategy to protect and recover the genetic diversity of C. forrestii and a further comprehensive study of its ecological traits in the future.

Elucidating the genetic background of threatened species is fundamental to their management and conservation, and investigating the demographic history of these species is helpful in the determination of the threats facing them. The woody species of the genus Magnolia (Magnoliaceae) have high economic, scientific and ecological values. Although nearly half of all Magnolia species have been evaluated as threatened, to date there has been no population genetic study employing Next Generation Sequencing (NGS) technology in this genus. In the present study, we investigate the conservation genomics of Magnolia fistulosa, a threatened species endemic to the limestone area along the Sino-Vietnamese border, using a double digest restriction-site-associated DNA-sequencing (ddRAD-seq) approach. To increase the reliability of our statistical inferences, we employed two approaches, Stacks and ipyrad, for SNP calling. A total of 15,272 and 18,960, respectively, putatively neutral SNPs were generated by Stacks and ipyrad. Relatively high genetic diversity and large population divergence were detected in M. fistulosa. Although higher absolute values were calculated using the ipyrad data set, the two data sets showed the same trends in genetic diversity (π, He), population differentiation (FST) and inbreeding coefficients (FIS). A change in the effective population size of M. fistulosa within the last 1 Ma was detected, including a population decline about 0.5–0.8 Ma ago, a bottleneck event about 0.2–0.3 Ma ago, population fluctuations during the last glacial stage, and the recovery of effective population size after the last glacial maximum. Our findings not only lay the foundation for the future conservation of this species, but also provide new insights into the evolutionary history of the genus Magnolia in southeastern Yunnan, China.

Long-term ecological and genetic studies in natural populations of tree species require marking techniques so that individuals can be re-visited over time, even in difficult terrain. Both GPS coordinates and physical labels have disadvantages that can make re-finding trees difficult. We tested passive and semi-active radio frequency identification (RFID) tags and readers as a means to relocate individual trees. Passive RFID tags do not provide a good solution because of low transmission power of hand-held readers and strong directionality. Semi-active RFID tags provide detection over longer distances, but also suffer from strong directionality. Active RFID tags promise an improvement over semi-passive tags, and could be evaluated in a future study. We conclude that RFID technology has the potential to improve the ability of researchers to locate individual trees repeatedly under natural conditions, and can be used in conjunction with other marking techniques such as physical tags and GPS coordinates.

Long-term ecological and genetic studies in natural populations of tree species require marking techniques so that individuals can be re-visited over time, even in difficult terrain. Both GPS coordinates and physical labels have disadvantages that can make re-finding trees difficult. We tested passive and semi-active radio frequency identification (RFID) tags and readers as a means to relocate individual trees. Passive RFID tags do not provide a good solution because of low transmission power of hand-held readers and strong directionality. Semi-active RFID tags provide detection over longer distances, but also suffer from strong directionality. Active RFID tags promise an improvement over semi-passive tags, and could be evaluated in a future study. We conclude that RFID technology has the potential to improve the ability of researchers to locate individual trees repeatedly under natural conditions, and can be used in conjunction with other marking techniques such as physical tags and GPS coordinates. [ABSTRACT FROM AUTHOR]