Floristic Characteristics of the Rare and Endangered Plants and the Conservation Countermeasures in Tibet, China

The floristic elements and the geographical distribution are analyzed in thispaper based on statistics of elements of rare and endangered plants in Tibet. The results have beengained as following: (1) According to 'the National Important Wild Conservative Plants List (List1)' and 'the National Important Wild Conservative Plants List (List 1)', there are a total of 54plant species (48 genera and 33 families); (2) The geographical elements are very complicated inTibet with 12 of 15 distribution patterns of genera classified by academician Wu; (3) There areobvious temperate genera with 28 genera accounting for 60. 4% of the total genera; (4) There areabundant endemic species accounting for 18. 52% of total species but poor endemic genera; (5) Thegeographical distribution is uneven and a great of species distribute in the areas between 1 000 mand 3 500 m above sea level; (6) To protect the rare and endangered plants efficiently, sixconservation measures are proposed, and 35 species are suggested for the conservative plants of theautonomous conservation level.

Leaf phenotypic variation of endangered plant Tetracentron sinense Oliv.and influence of geographical and climatic factors

To analyze the degree and pattern of phenotypic variation in leaves of Tetracentron sinense Oliv from the perspective of genetic and environmental adaptation and thus contribute to effective evidence-based conservation and management strategies for germplasm resources,we measured 17 morphological and epidermal micromorphological leaf traits from 24 natural populations of T.sinense.Nested analysis of variance,multiple comparison,principal component analysis(PCA),cluster analysis,and correlation analysis were used to explore phenotypic leaf variation among and within populations and potential correlations with geographic and environmental factors.There were significant differences in 17 leaf phenotypic traits among and within populations.The mean phenotypic differentiation coefficient of the 17 traits was 56.34%,and the variation among populations(36.4%)was greater than that within populations(27.2%).The coefficient of variation(CV)of each trait ranged from 4.6 to 23.8%,and the mean was 11.8%.Phenotypic variation of leaves was related to environmental factors such as average annual sunshine hours,average July temperature,and average annual rainfall.The variation changed along gradients of longitude,latitude,and altitude.The PCA clustered the 24 natural populations into four groups.Our study suggests that phenotypic variation in T.sinense occurred primarily among populations,with moderate levels of phenotypic differentiation among populations and low levels of phenotypic variation within populations.The plant’s poor adaptability to the environment is likely an important contributor to its endangerment.Accordingly,conservation strategies are proposed to protect and manage the natural populations of T.sinense.

Distribution and Current State of Rare and Endangered Tulips (Liliaceae) Arid Zones of Uzbekistan

Central Asia is considered to be the primary center of origin and diversity for the genus Tulipa L. (Liliaceae). The article considers the species of rare and endemic Tulipa L. species in the arid regions of Uzbekistan. There are nine registered species of the genus Tulipa. In the course of the study, the demographic indicators of the modern population of 39 populations were evaluated. 39 populations of the studied species were identified, most of which grow in the composition of the wormwood formations characteristic of the Turanian floristic province.

Somatic Embryogenesis and Organogenesis for Regeneration of Endangered Multipurpose Desert Plant <i>Leptadenia pyrotechnica</i>Forsk. Decne in the Kingdom of Bahrain

Leptadenia pyrotechnica is an important multipurpose endangered plant in the Kingdom of Bahrain with restricted distribution. Nodal explants were cultured on Murashige and Skoog (MS) medium supplemented with different concentrations and combinations of indole acetic acid (IAA) and 6-benzylaminopurine (BAP). Initially, 80% and 60% explants responded in direct shoot and callus initiation response respectively in presence of 8.88 μM BAP with 5.71 μM IAA in modified MS media after two weeks of culture. The highest frequency of plant regeneration was observed in presence of 8.88 μM BAP with 1.14 μM IAA following organogenic pathway of differentiation. Hundred percent callus proliferation was observed while initial callus developed in presence of 4.44 μM BAP with 2.85 μM IAA and was transferred in media containing 4.44 μM, 6.66 μM BAP with 2.85 μM IAA and 13.32 μM BAP with 5.71 μM IAA. The callus derived plants were regenerated following the pathway of indirect somatic embryogenesis. The induction of somatic embryogenesis and plant regeneration from callus was also observed in modified MS media supplemented with 4.44 μM BAP and 2.85 μM IAA. The plant regeneration protocol we developed for Leptadenia pyrotechnica will be very beneficial for biodiversity conservation and environment protection of Bahrain. Moreover, the present paper reports for the first time specifically the somatic embryogenesis in this multipurpose desert plant Leptadenia pyrotechnica.

Plants Diversity of the Burigi-Chato National Park: Rare and Invasive Species

This study was conducted within a distance of three to five kilometres around Lake Burigi as a study area in the Burigi-Chato National Park for the purpose of documenting the diversity of plants, identifying invasive and rare species for enhancing conservation in the park. Lake Burigi was purposively sampled as a study area for plant identification due to its potential as a tourist destination. No study has been conducted in the study area to reveal plants diversity, with a focus on documenting invasive and rare species, thus necessitating undertaking of this study to inform the park authority to take urgent control measures in avoiding widespread of invasive species and proper planning for conservation of rare species. The methods involved included field observation;plant identification using field guide books, indigenous and professional knowledge;field mapping using GPS receiver and literature review. Analysis of meteorological data and soil sample were also used among other methods. Meteorological data were analysed using Microsoft Excel and SPSS v.20 to determine the relationship between variables. A total of 102 plant species of different growth forms of trees, shrubs, grasses, sedges, forbs, and herbs were identified and recorded. Tegetes minuta and Argemone mexicana were recorded as invasive plant species, while six rare plant species potential for pharmaceutical industry;Zanthoxylum usambarense, Gardenia ternifolia, Faidherbia albida, Harrisonia abyssinica, Anona senegalensis, and, Pappea capensis were also recorded. Scars of wildfires were observed in the study area. The study area received an average rainfall of 964.36 mm per year, the highest peak recorded in 1951. The trend of rainfall showed that many years had rainfall below the average while the temperature was found to increase from year to year, the situation which suggests the existence of climate change in the study area. There was a weak negative relationship between temperature and wind speed. Soil nutrients and disturbances in the area were found to favour the growth of Tegetes minuta. Findings of this study would help ecological unit in the park to conduct regular ecological assessment for the purpose of controlling invasive plant species, which if left to flourish are likely to reduce habitat suitability for ungulates to utilize the area. Moreover, knowing which plant species are rare, adds value to the area as a destination to visitors interested in plants. The findings also allow the park authority to keep record of species rarity and thus easily take control of them to avoid unintended exploitation. Based on the findings, it was concluded that a number of disturbances, meteorological and edaphic factors favour the growth of invasive species. Urgent measures are to be taken to control the observed invasive plant species before they are left to spread in the park to avoid destruction of the habitat suitability and incurring unnecessary expenses and time in combating them. Furthermore, rare plant species especially those with pharmaceutical and industrial values are to be conserved with special attention to ensure their continued survival. It was recommended that human-induced activities should be prevented through regular ecological assessment, field patrols and public awareness programs. A similar study needs to be conducted during dry season to make comparison of the capacity of plants in adapting to different changes of the weather.

Rapid <i>in Vitro</i>Multiplication of an Endangered Medicinal Plant Sarpgandha (<i>Rauwolfia serpentina</i>)

An efficient and reproducible procedure for clonal multiplication through in vitro culture of Rauwolfia serpentina (L.) Benth. (Sarpagandha) is standardized. The different explants like leaf and stem were used for callus induction and regeneration of the complete plantlets. High-frequency callusing was induced in leaf and stem explant of Rauwolfia serpentina on modified Murashige and Skoog (MS) medium supplemented with 2.5 mg/l 2,4-D. Maximum regeneration of shoots from callus (90%) was observed in MS medium supplemented with 0.2 mg/l NAA and 1.5 mg/l BA. However direct regeneration (96%) was recorded best in MS medium supplemented with BAP 2.5 mg/l. The maximum number of shoots per explant (6.5) was also highest in this phyto-hormone combination. Higher induction of root (100%) was observed in MS medium supplemented with NAA 0.5 mg/l. The rooted plantlets were successfully established in the field.

Morphological Diversity and Nomenclature of Swertia chirayita (Gentianaceae)—Recovery of Endangered Medicinal Plant Population in North Eastern Himalaya

Swertia chirayita is a highly endangered and very well known medicinal plant enlisted in the IUCN list of endangered plants in the world. The plant is currently having a very low population in eastern Himalayan region due to its very low germinability in its own natural habitat. The plant population was found to have a range of diversity in morphological observations and some of the morphovariants were found to be very low among the variant populations which created a concern due to its diminishing range of genetic base in this region. A basic group of 5 morphovariants and 24 sub variants were identified and separately maintained. It is a pluriannual plant and reproductive or flowering stage was observed after completion of two years of vegetative stage from the time of seed germination. A range of morphovariant diversity was observed in the populations of Neora Valley and Lava region of eastern Himalaya in our investigation. Diversity was critically observed in some of the parameters of different morphovariants in the population. Conservation practices were generally done by not making groups or sub groups of this endangered plant while we proposed the way of conservation by making groups for proper conservation of this vulnerable plant.

Review of Studies on Rare Earth against Plant Disease

Agricultural application of rare earth (RE) has been generalized for several decades, and it is involved in crops, vegetables and stock raising in China. However, all the researches on RE mainly focus on the fields such as plant physiological activity, physiological and biochemical mechanism, sanitation toxicology and environmental security. Plant protection by using RE and the induced resistance of plant against diseases were summarized. The mechanism of rare earth against plant disease is highlighted, which includes following two aspects. First, RE elements can control some phytopathogen directly and reduce its virulence to host plant. Another possibility is that RE elements can affect host plant and induce the plant to produce some resistance to disease.

A review of fractionations of rare earth elements in plants

Studies were carried out on several aspects of rare earth elements （REEs）, such as the theory and practice of their applications in agriculture, their geochemical behaviors in natural and agricultural ecosystems, the mechanisms for the increase of crop yield using REE fertilizer, and their toxicology. However, limited knowledge was available for the transfer processes and the features and mechanisms of distribution and fractionations of REEs inside plants. The characteristics of REE fractionations in plants can be used to ＂trace＂ the pathway of REE transportation from soils （solution） to plants. A better understanding of the mechanisms of REE fractionations was helpful to investigate the controlling factors, including both the internal and the external ones. The characteristics and mechanisms of REE fractionations in plants and their significance were reviewed. Furthermore, the prospect for these fields was discussed, in hope of providing a new way in studying the bioavailability of REEs and heavy metals.

Effects of plant growth regulators, carbon sources and pH values on callus induction in Aquilaria malaccensis leaf explants and characteristics of the resultant calli

The endangered tropical tree, Aquilaria malaccensis, produces agarwood for use in fragrance and medicines. Efforts are currently un-derway to produce valuable agarwood compoundsn tissue culture. The purpose of this study was to develop an optimal growth medium, specif-ically, the best hormone combination for callus suspension culture. Using nursery-grown A. malaccensis, sterilized leaf explants were first incu-bated on basic Murashige and Skoog （MS） gel medium containing 15g/L sucrose and at pH 5.7. Different auxin types including 1-naphthaleneacetic acid （NAA）, 2,4-dichlorophenoxyacetic acid （2,4-D）, and indole-3-butyric acid （IBA）, were tested at various concentrations （0.55, 1.1 and 1.65 μM） using the basic medium. Leaf explants were incubated for 30 days in the dark. Callus induced by 1.1 μM NAA had the highest biomass dry weight （DW） of 17.3 mg;however the callus was of a compact type. This auxin concentration was then combined with either 6-benzylaminopurine （BAP） or kinetin at 0.55, 1.1, 2.2 or 3.3 μM to induce growth of friable callus. The 1.1μM NAA＋2.2μM BAP com-bination produced friable callus with the highest biomass （93.3mg DW）. When testing the different carbon sources and pHs, sucrose at 15g/L and pH at 5.7 yielded highest biomasses at 87.7mg and 83mg DW, respec-tively. Microscopic observations revealed the arrangement of the friable cells as loosely packed with relatively large cells, while for the compact callus, the cells were small and densely packed. We concluded that MS medium containing 15 g/L sucrose, 1.1 μM NAA ＋ 2.2 μM BAP hor-mone combination, and a pH of 5.7 was highly effective for inducing friable callus from leaf explants of A. malaccensis for the purpose of establishing cell suspension culture.

Adaptive introductions:How multiple experiments and comparisons to wild populations provide insights into requirements for long-term introduction success of an endangered shrub

Recovery of an imperiled plant species may require augmentation of existing populations or creation of new ones.Hundreds of such projects have been conducted over the last few decades,but there is a bias in the literature favoring successes over failures.In this paper,we evaluate a series of introductions that experimentally manipulated microhabitat and fire in an adaptive introduction framework.Between 2002 and 2012,we(and our collaborators) carried out ten introductions and augmentations of Florida ziziphus Pseudoziziphus(Condalia,Ziziphus) celata,a clonal shrub limited to very small populations and narrowly endemic to pyrogenic central Florida sandhills.Six of the introductions were designed as experiments to test hypotheses about how demographic performance was affected by microhabitat,fire,and propagule type.Introduced transplants had high survival(<90%annually),inconsistent growth,and little transition to reproduction,while introduced seeds had low germination and survival.Transplants were more efficient than seeds as translocation propagules.Shaded(vs.open) sites supported generally higher transplant and seedling survival and seed germination percentages,but growth responses varied among experiments.Supplemental irrigation increased transplant survival and seed germination,but otherwise seedling and plant survival and growth were not significantly affected.Contrary to expectations based on wild populations,introduced propagules have not been more successful in unshaded sites,suggesting that Florida ziziphus has broader microhabitat preferences than hypothesized.Compared to wild plants,introduced plants had similar survival and responses to fire,slower growth,and more delayed flowering.Introduced plants had no clonal spread.While no introduced population has demonstrated a capacity for long-term viability,one augmented population has flowered and produced viable fruits.Given that Florida ziziphus genets are long-lived,low levels of sexual reproduction may be adequate for the establishment of viable populations.Thus,after many translocations over more than a decade,it is premature to characterize any single translocation as a success or a failure,underscoring the need for a long view of translocation success.

Habitat suitability and herbivores determine reintroduction success of an endangered legume

Reintroductions of rare plants require detailed knowledge of habitat requirements, species interactions,and restoration techniques. Thus, incremental experimentation over many years may be required to develop adequate knowledge and techniques for successful reintroduction. To determine drivers of extinction in historical reintroductions of a federally endangered perennial(Astragalus bibullatus), we developed a reintroduction experiment to disentangle the relative importance of habitat quality, herbivores, and restoration technique on reintroduction success. In a factorial design, we manipulated access to vertebrate herbivores across different habitat types(mesic ecotone vs. xeric barren), and used founder populations comprised of more transplants and genetic sources than previous reintroduction attempts.In mesic ecotones where historical reintroductions failed, excluding herbivores, thinning woody encroachment to improve habitat quality, outplanting across a greater array of microhabitats, and increasing founder population size did not improve demographic rates over previous attempts.Compared to mesic ecotones, transplant survival rates and cumulative fruit production were more than two and ten times greater, respectively, in a xeric barren ecotone characterized by open, grassy, and dry microenvironmental conditions. Across all sites, herbivores decreased probabilities of survival and flowering of larger adult plants. Flowering rates were 80% greater inside relative to outside herbivore exclusion cages. Over a four-year period, only a single uncaged plant produced fruit. Our study demonstrates that habitat quality and vertebrate herbivory are key drivers of long-term persistence in rare plant reintroductions. Using incremental experiments that build on previous knowledge gained from long-term monitoring can improve reintroduction outcomes.

Mating system and genetic diversity of a rare desert legume Ammopiptanthus nanus (Leguminosae)

Ammopiptanthus nanus is an endangered evergreen shrub endemic to the deserts of central Asia and plays an important role in delaying further desertification. We examined allozyme variation and AFLP diversity in A. nanus populations and investigated the mating system of this species using progeny arrays assayed for polymorphic allozyme loci. Mating system analysis in the Keyi＇eryongke＇er population showed low levels of outcrossing, and strong inbreeding depression. Low levels of genetic variation were detected at both population （allozyme, Pp=14.0%, A=l.14, He=0.031; AFLP, Pp=14.5%, Shannon＇s information index I=0.063） and species （allozyme, Pp=21.1%, A=1.21, He=0.040; AFLP, Pp=20.9%, I=0.083） levels; while moderate genetic differentiation existed among populations, as indicated by allozymes （Gsa-=0.081） and AFLP （GST=0.151-0.193）. Founder effect, bottlenecks in evolutionary history, the mixed mating system and co-ancestry may have influenced the level of genetic diversity in A. nanus. Markers of both types provide new insights for conservation management, indicating that the Biao＇ertuokuoyi and Keyi＇eryongke＇er populations should be given priority for in situ conservation and regarded as seed sources for ex situ conservation.

Age structure and static life tables of the endangered Juniperus phoenicea L. in North Sinai Mountains, Egypt:implication for conservation

Studying the population ecology of endangered plants provides important baseline information for its monitoring and conservation.Juniperus phoenicea L.is an endangered species in arid ecosystems in Africa and the Middle East.The static life tables,survival curve and age structure of J.phoenicea populations from two mountains in North Sinai,Egypt(Gabal(G.)El-Halal and G.El-Maghara)were investigated.In each mountain,fifteen plots were selected,and field measurements such as stand density,tree height,and crown diameter were conducted.Moreover,44 trees were cored and crossdated according to standard dendrochronological procedures.The results showed that the tree ages ranged from 50 to 262 years at G.El-Halal and 96 to431 years at G.El-Maghara.Mature J.phoenicea individuals dominated the study area,with only a few individuals being younger than 100 years.Moreover,seedling recruitment was extremely limited.Since the studied J.phoenicea populations showed high mortality rates among both old and young individuals,there is an imminent need for establishing a conservation program to prevent its extinction in the future.Therefore,management and conservation efforts should be made to minimize human disturbance and protect the relic habitats of this endangered species at its southern distribution limits in Africa.

In-situ Protection Management and Conservation Study of Some Medicinal Plants

In-situ study on eleven commercially important species viz; Adiantum capillus-veneris L, Bergenia ciliata (Haw) Sternb, Colchicum luteum Baker, Polygonum amplexicaule D.Den, Cuminum cyminum L, Dioscorea deltoidea Wall Kunth,L Morchella esculenta L, Paeonia emodi Wall ex H Kf, Podophyllum hexandrum (Royle) Chatt & Mukh, Valeriana wallichii DC and Viola serpens Wall ex Roxb was conducted in four locations viz; Malam (1 400 to 2 000?m),Bargin (1 700 to 2 300?m),Biakand (1 500 to 2 100?m) and Shinko (2 100 to 2 700?m).The data was recorded from both protected and unprotected sites of each site.Each site had 3 altitudinal sampling point.The density, herbage coverage and fresh biomass were determined in each locations for every species.All the investigated parameters generally showed an increase of 3 to 6 times over unprotected sites in each locations. Morchella esculenta,Dioscorea deltoidea,Colchicum luteum and Podophyllum hexandrum were absent in all unprotected sites while other species had low values in these sites.The investigated parameters of Colchicum luteum,Bergenia ciliata,Paeonioa emodi,Dioscorea deltoidea and Podophyllum hexandrum generally increased with the increasing elevation.Soil analysis,soil and air temperatures were recorded for each site.The air and soil temperature were slightly higher in open areas than in the protected site and showed decrease with increasing elevation.While the soil fertility was relatively high in protected sites as compared to unprotected area.The study shows that protection promotes the growth, distribution and occurrence of medicinal plants.It is possible with the participation of local communities to conserve these resources.

Micropropagation and Callus Culture of Saussurea laniceps, an Alpine Medicinal Plant

Cottonhead windhairdaisy (Saussurea laniceps Hand.-Mazz.) is one of the most famous and important medicinal herbs in China. Illegal collection from wild populations is increasingly threatening the present environment of S. laniceps. Estab- lishment of an efficient method for micropropagation is the best way to change its endangered situation. When mature seeds of S. laniceps were cultured on hormone-free MS medium, plantlets were formed from germinated seeds in 7–10 d. Then 0.5 cm × 0.5 cm leaf explants were transplanted to MS medium supplemented with 1-naphthalene-acetic acid (NAA)/2,4-D and benzyladenine (BA)/KT and callus was achieved 10 d after transfer. Shoot bud regeneration occurred from callus cultured on MS medium supple- mented with different growth regulators 20 d after culturing. The regeneration percentages varied with the different components of plant growth regulators. The percent regeneration from callus pretreated at low temperature of 5°C increased significantly compared with those incubated at 23/20°C directly. Optimal regeneration was observed with explants on media supplemented with 1.5 mg·L–1 BA plus 0.2 mg·L–1 NAA. In the presence of 0.2 mg·L–1 NAA in half-strength MS, 78% of the shoots formed roots. Plantlets from explants showed 63% survival after acclimatization.

Floral Homeostasis Breakdown in Endangered Plant <i>Valeriana jatamansi</i>Jones (Valerianaceae) in North Eastern Himalayan Region

An inhabitant of north western Himalayan region and a gynodioceious plant having a lot of medicinal properties, Valeriana jatamansi Jones (Valerianaceae) is an enlisted endangered plant in the world. It was introduced in the research station of North Bengal Agricultural University (27.06&degN 88.47&degE) situated in north eastern Himalayan region of Darjeeling district in the state of West Bengal from Sikkim Sangtok (27&deg25'N 88&deg31'E) of north Sikkim area in India for the purpose of conservation. As we noticed some irregular development of floral organs, we investigated some pertinent questions regarding ecological aberrations found in plants. We found plants introduced in north eastern Himalayan region changed their homology of number flower petals and position of stamens seen naturally in north western Himalayan region as per the reports. Was there any genetic or extreme environmental stress condition caused a sudden change in floral structure as it is generally known as a rare phenomenon and frequently not seen? What were the correlations of different floral parts and fitness of population in different morphotypes? We predicted possible outcomes of seed setting by univariate regression models in a particular environment in addition to this investigation. We proposed three models of heterozygousity for answering the reasons of unstable floral form from a general known floral form, where silent mutations help the plants to survive in adverse conditions in spite of deformed or variable formed of floral morphology.

Assessment of Diversity, Distribution, Conservation Status and Preparation of Management Plan for Medicinal Plants in the Catchment Area of Parbati Hydroelectric Project Stage-Ⅲ in Northwestern Himalaya

The developmental activities, particularly the construction of hydroelectric projects are causing a great loss of biodiversity in the Indian Himalayan Region. The Himachal Pradesh, a part of IHR is well known for the development of hydroelectric projects. The Parbati H.E. Project is amongst the major projects of the State. The different stages of the project are all causing loss of biodiversity of the area. Stage Ⅲ of the Parbati H.E. Project is a run of the river scheme on the Sainj River downstream of Power House of Parbati H.E. Project Stage Ⅱ. The project shall utilize regulated discharge of Parbati H.E. Project Stage Ⅱ and inflow of River Sainj for power generation, and has been contemplated as a peaking station operating in tandem with Stage Ⅱ. The present study has been undertaken to see the impact of hydroelectric project on the biodiversity, particularly on medicinal plants. A total of 104 species of medicinal plants, belonging to different life forms, i.e., trees (23 spp.), shrubs (22 spp.), herbs (57 spp.) and ferns (2 spp.) were recorded. The species have been analyzed and studied for their distribution, classification, altitudinal zones, part (s) used, indigenous uses, nativity, endemism and rarity.Different parts of these species, such as whole plants, roots (including rhizomes and tubers), leaves, flowers, fruits, seeds, stems, barks, spikes, nuts and insect galls are used by the inhabitants for curing various diseases and ailments. 30 species are native to the Himalayan region, 9 species native to the Himalayan region and adjacent countries also and 65 species are non-natives. 9 species are near endemics. Considering the whole Himalaya as a biogeographic unit (sensu lato), the near endemics are endemic to the Himalaya. Among these species, Zanthoxylum armatum is categorized as Endangered and Valeriana wallichii as Vulnerable. Hedychium spicatum, Rhus javanica, Berberis lycium, Thalictrum foliolossum, Salvia lanata, Rubia cordifolia and Bergenia ligulata may be considered as threatened species due to their over exploitation for trade. 90 species are propagated by seeds, 8 species by seeds and rhizomes/roots/tubers, 4 species by seeds and cuttings, and 2 species by sori. A management plan for the cultivation and conservation of the medicinal plants in the dam submergence area, and the commercially viable medicinal plants with high value in the catchment area is suggested.

Ecological features of Aconitum balfourii (Bruhl) Muk.-an endangered medicinal plant in the northwest Himalaya

Aconitum balfourii （Bruhl） Muk. （Ranunculaceae） is an en- dangered medicinal plant. Natural populations were surveyed in the northwest Himalaya for population estimation and evaluation of elite germplasm. Vegetation sampling of quadrats was conducted using the vertical belt transect method in areas supporting A. balfourii. Plant den- sity and relative dominance ofA. balfourii were low compared with other alpine species. Threat status was determined on a site-by-site basis as well as for entire western Himalaya..4. balfourii was generally classified as endangered on the basis of geographic distribution and critically en- dangered on the basis of abundance. We found the natural geographic distribution of.4. balfourii to be diminishing due to habitat destruction. Immediate corrective management measures are needed for sustainable utilization and long-term conservation of the species in the wild.

Development and characterization of 43 microsatellite markers for the critically endangered primrose Primula reinii using MiSeq sequencing

Primula reinii(Primulaceae), a perennial herb belonging to the Primula section Reinii, occurs on wet,shaded rocky cliffs in the mountains of Japan. This threatened species comprises four varieties; these plants are very localized and rare in the wild. In this study, 43 microsatellite markers were developed using MiSeq sequencing to facilitate conservation genetics of these critically endangered primroses. We developed novel microsatellite markers for three varieties of P. reinii, and tested its polymorphism and genetic diversity using natural populations. These novel markers displayed relatively high polymorphism; the number of alleles and expected heterozygosities ranged from 2 to 6(mean = 3.2) and 0.13 to 0.82(mean = 0.45), respectively. All loci were in Hardye Weinberg equilibrium. These microsatellite markers will be powerful tools to assess P. reinii genetic diversity and develop effective conservation and management strategies.