Zeuxine gracilis, Zeuxine strateumatica, and Eulophia sinensis are wild orchids with different breeding systems and colonizing abilities. Zeuxine gracilis is an outcrosser with restricted distribution, whereas S. strateumatica is an apomictic colonizer found only in newly available open habitats. Eulophia sinensis is an outcrossing colonizer. This study investigates the levels of genetic variation and patterns of population structure in these wild orchids to provide genetic information for the development of suitable conservation strategies. Lack of allozyme variation was characteristic of all three species, especially in populations of the two colonizing orchids, Z. strateumatica and E. sinensis. More variable markers, randomly amplified polymorphic DNAs (RAPDs), were further employed to characterize population structure of these species. Substantial genetic variation was found at the RAPD loci within populations of Z. gracilis (p = 21.65 ± 15.88%, A = 1.217 ± 0.159, and H = 0.076 ± 0.054) and E. sinensis (p = 17.82 ± 20.97%, A = 1.179 ± 0.209, and H = 0.070 ± 0.084), but little variation existed within populations of Z. strateumatica (p = 2.84 ± 2.58%, A = 1.029 ± 0.026, and H = 0.011 ± 0.011). Regardless of the breeding system, the total gene diversity at the species level was partitioned primarily between populations, as shown by high G(ST) values, in all three species. An extremely high level of population differentiation (G(ST) = 0.924) was found in the apomictic colonizer Z. strateumatica. The patterns of genetic variation in these wild orchids are apparently related to their differences in breeding system and colonizing ability. Different conservation strategies are needed for the long-term survival of these species.
Goodyera procera is an endangered terrestrial orchid in Hong Kong. Information on its reproductive biology and pattern of genetic variation is needed to develop efficient conservation strategies. Pollination experiments showed that the species is self-compatible, but dependent on pollinators for fruit set. Bagged plants produced no fruits. Artificial pollinations resulted in 92% fruit set through selfing, 94% with geitonogamous pollination, and 95% following xenogamous pollination. Fruit set in the open-pollinated control was 75% at the same sites. Allozyme electrophoresis and random amplified polymorphic DNA (RAPD) were used to evaluate genetic variation and structure of 15 populations of Goodyera procera. Despite its outbreeding system, allozyme data revealed low variation both at the population (P = 21.78%, A = 1.22, and H = 0.073) and species (P = 33%, A = 1.33, and H = 0.15) levels, in comparison with other animal-pollinated outbreeding plant species. However, RAPD variation was relatively high (P = 55.13% and H = 0.18 at the population level, and P = 97.03% and H = 0.29 at the species level). G(ST) estimates indicated high levels of genetic differentiation among populations (G(ST) = 0.52 and I = 0.909 ± 0.049 based on allozyme data, and G(ST) = 0.39 and I = 0.859 ± 0.038 based on RAPD data), much above the average for outcrossing species, suggesting that gene flow was limited in this species. Based on these data, suitable strategies were developed for the genetic conservation and management of the species.
The pollination biology, mating system, and population genetic structure of Kandelia candel were investigated. Field observations on its pollination and reproductive biology suggested that this species is pollinator dependent for fruit set, and bee activities can lead to substantial geitonogamous selfing. Quantitative analysis of the mating system parameters was performed using progeny arrays assayed for allozyme markers. Multilocus outcrossing rates (t(m)) were estimated to be 0.697 ± 0.091 and 0.797 ± 0.062 in two populations. In comparison to other plant species with mixed-mating system, the level of allozyme variation was very low in the 13 populations sampled along the coastlines of Hong Kong. At the species level, the proportion of polymorphic loci was 20%, number of alleles per locus was 1.2, and heterozygosity was 0.0362. The total gene diversity was primarily distributed within populations (H(S )= 0.0339), and the coefficient of genetic differentiation among populations was low (G(ST )= 0.064). This pattern of population genetic structure suggests that gene flow, primarily in the form of water-dispersed seedlings in viviparous mangrove species, is not as limited as previously thought. However, microgeographic pattern in allele frequency at the marker loci could still be detected between the western and eastern coastal populations.
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