Genetic Diversity of Cephalotaxus mannii, a Rare and Endangered Plant

Using RAPD technique, the DNA diversity of Cephalotaxus mannii Hook. f., its genetic diversity pattern, the reasons for its endangered position and conservative approaches were studied. The results show that: 1. The genetic diversity of C. mannii collected from five localities in Hainan is low, and its adaptability to environmental change is weak. 2. The differences of genetic diversity between intra- and inter-populations are great, and the major variation distributes within the population (DNA diversity is 85.1%). 3. The excessive lumbering, man-made destruction, violent typhoon, edible value of the seeds and genetic drift were the main reasons for the low-level genetic diversity of C. mannii and its endangered position. 4. The difference of the micro-environment and other random factors affecting the population should also be taken into full consideration in the study and in protection of such occasionally scattered plants. 5. Enforced measures should be taken to protect the present population, enlarge the population and lower the loss rate of its gene. Mt. Limulin should be chosen as a conservative spot because of its high genetic diversity and less destruction of the forest. Meanwhile, the protection of other populations should be enforced. 6. The differences within and between the populations are great based on different primers used. The change of proportions in polymorphic loci between the populations is more than that between the primers.

Studies on the Reproductive Biology and Endangerment Mechanism of the Endangered Plant Manglietia aromatica

The embryogenesis, pollen germination, floral character and seed physiology of the endangered plant Manglietia aromatica Dandy were investigated. Based on this study, this species has very low seed set rate. The abortion rate of functional megaspores in all the ovules is 27.9%, the egg cell abortion rate of mature embryo sacs is up to 80%, and the germination rate of pollen grains is as low as nearly 0.01%. In addition, the floral structure appears to be another limited factor for the effective pollination of this species. The endangerment mechanism of this species seems to be comprehensive. Human's destroying actions are the direct factors that have made the population degenerate quickly; low reproductive ability and the destroyed environments are the main reasons that prevent the population from renovating and spreading. Therefore, the conservation measures suggested by this study are to research the breed technology, artificial population renovating, in situ conservation, and ex situ conservation.

Investigation on the Rare and Endangered Plants in the Fenhe Source

Fouty-eight species of rare and endanged plants in the Fenhe Source,belong to 21 families 37 genera.There were 1specie of national second-class protected plant and 3 species of national third-class plants;4 species of first-class protected plants,6 species of second-class protected plants and 14 species of third-class protected plants in Shanxi Province.On the basis of analyzing the endanged reasons,related protection countemeasures were put forward in this article.

Advance of New Researches on Tissue Culture of Rare and Endangered Plant Elaeagnus mollis

This review summarized the recent research achievements on the process of tissue culture of the rare and endangered plant Elaeagnus mollis,including sterilizing protocols of different explants derived from adult plants and seeds for tube germination,two types of plants regeneration（organ type and organ genesis type）,methods of rooting and transplanting,factors affecting culture,as well as browning and vitrification phenomena and avoiding measures.And the further biotechnology research fields of E.mollis were prospected.

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.

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.

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.

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.

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.

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.

Efficient in vitro Propagation of an Endangered Species Kolkwitzia amabilis Graebn

This study developed an efficient in vitro cultivation and propagation sys- tem for an endangered species Kolkwitzia amabilis using nodal segments as ex- plants. Multiple shoots were induced through axillary bud formation. The highest fre- quency of multiple shoot induction was achieved when the nodal segment explants were incubated in Murashige and Skoog （MS） medium supplemented with 4.44 pM 6-benzyladenine （6-BA） in combination with 0.54 μM a-naphthaleneacetic acid （NAA）, followed by treatment with 4.44 μM 6-BA in combination with 0.27 μM NAA. Shoot multiplication could be induced in MS medium supplemented with stand-alone 6-BA or 6-BA in combination with indole-3-acetic acid （1.71 μM） or NAA （0.27 or 0.54 μM）, with 6-BA and either compound, exhibiting a stronger effect on shoot multiplication. The optimum combination of plant growth regulators for shoot multiplication was 4.44 μM 6-BA with 0.27 μM NAA. The maximum rooting percentage was obtained in a half-strength MS medium supplemented with indole-3-butyric acid alone and in com- bination with NAA and 2,4-dichlorophenoxyacetic acid, but the best combination of plant growth regulators for rooting was 1.48 μM indole-3-butyric acid with 1.08 μM NAA and 0.05 μM 2,4-dichlorophenoxyacetic acid. The rooted shoots were trans- ferred to a greenhouse with a success rate of 100%.

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.

Genetic effects of historical anthropogenic disturbance on a long-lived endangered tropical tree Vatica mangachapoi

The endangered Vatica mangachapoi,a longlived,tropical tree with economic and ecological importance found in Hainan,China,was used to assess the hypothesis that historical human activities in Hainan’s tropical rain forest could have negative effects on the genetic diversity of V.mangachapoi.Three hundred and twenty individuals from 11 natural populations—which were classified into three groups according to levels of disturbance—were sampled and analyzed with ISSRmarkers.Although genetic diversity of V.mangachapoi is high at the species level,it is relatively low within populations.A significant genetic differentiation occurs among different disturbance levels.Significant isolation-by-distance indicated relevant historical anthropogenic changes.Our findings showed that historical human disturbances significantly increase the genetic differentiation and slightly decrease the genetic diversity of long-lived tree V.mangachapoi.Relevant targeting conservation actions were recommended.

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.

Assessment of Reproductive Potential of Different Populations of Angelica glauca Edgew.,a Critically Endangered Himalayan Medicinal Herb

Angelica glauca is one of the important medicinal plants and it is widely used by indigenous communities for different purposes. The present study analyzes variability in reproductive characters of A. glauca. The reproductive parts were found having significant positive correlation with altitude (e.g.,number of umbellets /umber r= 0.857,p<0.05; umbel diameter r=0.735,p<0.05).

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.

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.

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 Himaehal 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. 3o 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 biogeographie unit （sensu lato）, the near endemics are endemic to the Himalaya. Among these species, Zanthoxylum armature 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.

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.