Assessment of genetic variation in Sicilian Helichrysum (Asteraceae) and implication to germplasm conservation

Abstract

Some taxa of Sicilian Helichrysum related to sect. Stoeachadina have an uncertain systematic position; moreover, they are rare and endangered. In order to preserve these taxa in a genebank, a strategy to keep their genetic variation as high as possible is required. The high informative content of AFLP markers was used to assess genetic variations within and between populations and their relationships. The results show that all populations are genetically similar, even though there are two distinct groups: the first is the population referred to H. pendulum, and the second is represented by other populations belonging to several different specific and intraspecific taxa. The H. pendulum population is homogeneous, and characterised by the lowest genetic variation among all the populations considered. The second group shows a greater variation as evidenced by the presence of three subgroups: (i) populations referred to H. panormitanum var. panormitanum and H. panormitanum var. stramineum, (ii) populations referred to H. errerae var. errerae and H. errerae var. messerii, (iii) a population referred to H. hyblaeum, which, despite morphological and chorological differences, is similar to the other populations of the second group from a genetic stand point. Each population of the above-mentioned taxa has local fragment markers whose frequency is useful to determine how many plants should be sampled in order to collect the seeds necessary to preserve the population's genetic diversity in a genebank.     

Contribution to a Taxonomic Revision of the Sicilian Helichrysum Taxa by PCA Analysis of Their Essential‐Oil Compositions

The chemical profile of the essential oils in ten populations of the genus Helichrysum Mill . (Asteraceae), collected in the loci classici of the nomenclatural types of the taxa endemic to Sicily, were analyzed. Our results confirm that the analysis of secondary metabolites can be used to fingerprint wild populations of Helichrysum, the chemical profiles being coherent with the systematic arrangement of the investigated populations in three main clusters, referring to the aggregates of H. stoechas, H. rupestre, and H. italicum, all belonging to the section Stoechadina. The correct nomenclatural designation of the investigated populations is discussed and the following two new combinations are proposed: Helichrysum preslianum subsp. compactum (Guss .) Maggio, Bruno, Guarino, Senatore & Ilardi and Helichrysum panormitanum subsp. latifolium Maggio, Bruno, Guarino, Senatore & Ilardi .     

Cytogenetic effects of nine <Emphasis Type="Italic">Helichrysum</Emphasis> taxa in human lymphocytes culture

Helichrysum Mill. (Asteraceae) species have been used in folk medicine for thousands of years in the world. The in vitro cytogenetic effects in human lymphocytes of nine Helichrysum taxa used in Turkey folk medicine were investigated. Blood samples were obtained from healthy donors, non-smoking volunteers, which were incubated and exposed to increasing concentrations of methanol extracts of Helichrysum taxa (0.01, 0.05, 0.1, 0.5 and 1 mg/mL). The inhibitory effects of H. stoechas (L.) Moench subsp. barrelieri (Ten.) Nyman, H. armenium DC. subsp. armenium, H. armenium DC. subsp. araxinum (Kirp.) Takht., H. plicatum DC. subsp. plicatum, H. compactum Boiss. and H. artvinense P.H.Davis & Kupicha on the mitotic index and replication index indicate that these taxa can have genotoxic and mutagenic effects. They should therefore not be used freely in alternative medicine although their antiproliferative activity may suggest anticarcinogenic properties. Increase effects of H. stoechas subsp. barrelieri, H. armenium subsp. armenium, H. armenium subsp. araxinum, H. chasmolycicum P.H.Davis, H. plicatum subsp. plicatum, H. compactum and H. artvinense on the micronucleus rates showed that these taxa can have genotoxic and carcinogenic effects.     

Genetic variation in Mediterranean Helichrysum italicum (Asteraceae; Gnaphalieae): do disjunct populations of subsp. microphyllum have a common origin?

The yellow‐flowered everlasting daisy Helichrysum italicum (Asteraceae, Gnaphalieae) is widely distributed in the Mediterranean basin, where it grows in continuous and widespread populations in diverse open habitats. Helichrysum italicum subsp. microphyllum has a disjunct distribution in the Balearic Islands (Majorca and Dragonera), Corsica, Sardinia, Crete and Cyprus. Numerous morphological intermediates between subsp. italicum and subsp. microphyllum are known from Corsica, where the two subspecies co‐occur. The aims of the study were to investigate if subsp. microphyllum has a common origin, constituting an independent gene pool from subsp. italicum, or if the morphological differences between subsp. microphyllum and subsp. italicum have arisen independently in different locations from a common wider gene pool. Our analyses of AFLP, cpDNA sequences and morphological characters show that there is geographic structure to the genetic variation within H. italicum, with eastern and western Mediterranean groups, which do not correspond with the division into subsp. microphyllum and subsp. italicum as currently circumscribed. Local selection on quantitative trait loci provides sufficient explanation for the morphological divergence observed and is consistent with genetic data. Within the western Mediterranean group of the species we found considerable polymorphism in chloroplast DNA sequences among and within some populations. Comparison with chloroplast DNA sequences from other Helichrysum species showed that some chloroplast haplotypes are shared across species.     

The allopolyploid origin ofSedum rupestre subsp.rupestre (Crassulaceae)

InSedum rupestre L. a polyploid series (x = 16) occurs in which aneuploid chromosome numbers and odd levels of ploidy prevail. The most common and widely distributed cytotype,S. rupestre subsp.rupestre, is 2n = 112. Plants resemblingS. rupestre subsp.rupestre can be obtained by hybridizing the tetraploid cytotypes ofS. forsterianum Sm. (2n = 48) andS. rupestre subsp.erectum 't Hart (2n = 64). Comparison of these artificial hybrids with their parents and a large number of plants ofS. rupestre subsp.rupestre (2n = 112) from nature showed thatS. rupestre subsp.rupestre and the artificial hybrids are morphologically indistinguishable, and intermediate betweenS. forsterianum andS. rupestre subsp.erectum. MorphologicallyS. rupestre subsp.rupestre is closer to subsp.erectum than toS. forsterianum. Chloroplast DNA restriction patterns ofS. rupestre subsp.rupestre, however, resembleS. forsterianum more closely. The combined results of the hybridization experiments, the analysis of the cpDNA restriction patterns, and the morphological variation indicate the allopolyploid origin ofS. rupestre subsp.rupestre. Natural hybrids inSedum (Crassulaceae) 4.     

Chloroplast DNA variation of Panax (Araliaceae) in Nepal and its taxonomic implications

The restriction site and size variation of five PCR amplified fragments of noncoding chloroplast DNA (cpDNA) was examined in material from 13 populations ofPanax from Nepal and China. Fourteen restriction endonucleases produced 81 restriction site and length variations from the large single-copy region of cpDNA, 27 of which are polymorphic. The cpDNA dataset suggests two distinct groups ofPanax from Nepal (clades I and II). Clade I consists of two populations ofP. pseudoginseng subsp.pseudoginseng, and clade II is composed of material referable toP. pseudogingeng subsp.himalaicus (vars.himalaicus, angustifolius, andbipinnatifidus). The three accessions ofP. pseudoginseng subsp.japonicus andP. ginseng studied from China had cpDNA characters that differed from the HimalayanPanax. The highly distinctive cpDNA profile and morphology ofP. pseudoginseng subsp.pseudoginseng sensu Hara (1970) from central Nepal support its status as a separate species, which has an extremely restricted distribution.     

Genetic structure of the endangered Leucomeris decora (Asteraceae) in China inferred from chloroplast and nuclear DNA markers

The genetic variation and structure of Leucomeris decora, an endangered species in China were investigated. Analyses of three chloroplast DNA (cpDNA) regions (the rpl16 intron, trnQ-5′rps16 intergenic spacer and rpl32-trnL intergenic spacer) and one nuclear gene (GAPDH: encoding glyceraldehyde 3-phosphate dehydrogenase) were conducted on 11 L. decora populations. Low levels of cpDNA genetic diversity were found in this species and within populations, with the identification of 2 haplotypes in a total of 2,745 bp, while the level of genetic diversity revealed by the nuclear gene GAPDH was relatively high, indicating that random losses of genetic polymorphisms from populations may have occurred recently. High levels of genetic differentiation among populations for both markers were detected in L. decora, which could be a consequence of the limited gene flow caused by geographic isolation among populations. An analysis of molecular variance revealed at the nuclear locus suggested the presence of geographic structure within the haplotype distribution possibly due to geographical barriers among populations. The haplotype network and mismatch distribution analyses did not detect the signal for a recent population expansion in L. decora. L. decora may persist in situ during climatic oscillations. Based on the genetic diversity and uniqueness of the populations, conservation strategies are discussed for this endangered species.     

Comparative phylogeographic study of Hosta sieboldiana and Hosta albomarginata (Asparagaceae) in Japan

We analyzed variations in chloroplast DNA (cpDNA) in the widespread herbaceous species Hosta sieboldiana and Hosta albomarginata across large portions of their geographic ranges in the Japanese archipelago. Our objective was to compare the phylogeographic histories and phylogeographic structures of the two congeneric species in the Japanese archipelago. The location of the study is Japanese archipelago. We sequenced 1380 bp of noncoding cpDNA from 45 populations of H. sieboldiana (n = 362) and 55 populations of H. albomarginata (n = 436) to assess genetic variations within and among populations across almost the entire distributions of the species in Japan. Extant patterns of geographic structure were analyzed using statistical parsimony networks and spatial analysis of molecular variance (SAMOVA). We also used Monmonier's algorithm to detect genetic barriers between regions. Relationships between the populations were examined using a neighbor‐joining (NJ) method. Four haplotypes were found for H. sieboldiana, whereas eight haplotypes were identified for H. albomarginata. Total genetic haplotype diversity (hT) and within‐population haplotype diversity (hS) for H. sieboldiana were 0.352 and 0.040, respectively, while the values for H. albomarginata were 0.529 and 0.085, respectively. The population differentiations (GST) for H. sieboldiana and H. albomarginata were 0.839 and 0.886, respectively. The SAMOVA analysis revealed two clusters in H. sieboldiana and four clusters in H. albomarginata. Differentiations between and among the clusters were supported by the BARRIER analysis and the NJ tree. We detected differences in the population genetic structure between the two species. We found that H. sieboldiana had lower haplotype diversity than H. albomarginata. These results may be partially explained by the difference in ecological habitats and geographic distributions between the species. Hosta albomarginata is more widely distributed than H. sieboldiana in East Asia including Russia, and this large distribution range would enable more chances to intraspecific gene flow.     

High‐throughput multiplex cpDNA resequencing clarifies the genetic diversity and genetic relationships among Brassica napus,Brassica rapa and Brassica oleracea

Brassica napus (rapeseed) is a recent allotetraploid plant and the second most important oilseed crop worldwide. The origin of B. napus and the genetic relationships with its diploid ancestor species remain largely unresolved. Here, chloroplast DNA (cpDNA) from 488 B. napus accessions of global origin, 139 B. rapa accessions and 49 B. oleracea accessions were populationally resequenced using Illumina Solexa sequencing technologies. The intraspecific cpDNA variants and their allelic frequencies were called genomewide and further validated via EcoTILLING analyses of the rpo region. The cpDNA of the current global B. napus population comprises more than 400 variants (SNPs and short InDels) and maintains one predominant haplotype (Bncp1). Whole‐genome resequencing of the cpDNA of Bncp1 haplotype eliminated its direct inheritance from any accession of the B. rapa or B. oleracea species. The distribution of the polymorphism information content (PIC) values for each variant demonstrated that B. napus has much lower cpDNA diversity than B. rapa; however, a vast majority of the wild and cultivated B. oleracea specimens appeared to share one same distinct cpDNA haplotype, in contrast to its wild C‐genome relatives. This finding suggests that the cpDNA of the three Brassica species is well differentiated. The predominant B. napus cpDNA haplotype may have originated from uninvestigated relatives or from interactions between cpDNA mutations and natural/artificial selection during speciation and evolution. These exhaustive data on variation in cpDNA would provide fundamental data for research on cpDNA and chloroplasts.     

Chloroplast DNA and isozyme evidence on the evolution ofSenecio vulgaris (Asteraceae)

Chloroplast DNA (cpDNA) and isozyme variation were analyzed over a range of populations of two infraspecific taxa of the tetraploidSenecio vulgaris. The isozyme data were supportive of the hypothesis that the weedy and cosmopolitanS. vulgaris var.vulgaris is an evolutionary derivative ofS. vulgaris subsp.denticulatus from the coasts of W Europe and montane altitudes in S Spain and Sicily. The two taxa exhibited a very high genetic identity with subsp.denticulatus containing slightly more isozyme diversity than was found in var.vulgaris. — Three cpDNA haplotypes (A, B, C) already known from other Mediterranean diploid species ofSenecio were resolved in var.vulgaris, and an additional fourth haplotype (E) was found in subsp.denticulatus. Two alternative hypotheses were chosen to account for the origin and maintenance of the observed cpDNA composition ofS. vulgaris. It either reflects (1) the retention of an ancestral polymorphism which stems from the recurrent and polytopic formation of ancestral tetraploid lineages; or (2)S. vulgaris originally was characterized by haplotype E, and haplotypes A, B and C were acquired through repeated introgressive hybridization with related diploid species. The finding that very low levels of nuclear (isozyme) diversity were present in both taxa ofS. vulgaris examined supports the second of these two hypotheses; however, more detailed analysis of nuclear genetic diversity is required before a firm conclusion can be reached on this matter.Dedicated to emer. Univ.-Prof. DrFriedrich Ehrendorfer on the occasion of his 70th birthday     

Chloroplast DNA variation and evolution in the genus Lens Mill

 Chloroplast DNA (cpDNA) restriction site diversity was assessed by 21 enzyme/probe combinations in 30 accessions of six Lens species, including the recently recognized L. lamottei and L. tomentosus. A total of 118 fragments were scored and 26 restriction site mutations were identified. The cpDNA restriction pattern supports circumscribing L. lamottei and L. tomentosus as independent species. The value of the data for reconstructing phylogeny in the genus is discussed. The cpDNA of all 13 accessions of the lentil’s wild progenitor, L. culinaris subsp. orientalis, differed from that of the single lentil cultivars used in this study. This diversity indicates that other populations of this subspecies from Turkey and Syria examined by Mayer and Soltis (1994) are potentially the founder members of lentil. Examination of L. lamottei×L. nigricans hybrids between accessions having different restriction patterns showed paternal plastid inheritance in L. nigricans. Received: 2 July 1996 / Accepted: 19 July 1996     

基于cpDNA单倍型多态性的草果栽培地理起源证据

为了探索草果(Amomum tsaoko)的栽培地理起源,该文检测了草果、拟草果(A. paratsaoko)的cpDNA序列变化,并获取了单倍型多态性信息。结果表明:(1)20个草果居群272个植株、5个拟草果居群62个植株共检测到7种单倍型。其中,草果有3种单倍型(H1、H3、H6),拟草果有6种单倍型(H1、H2、H3、H4、H5、H7)。H1和H3为共享单倍型,H6为草果私有单倍型,H2、H4、H5、H7为拟草果私有单倍型。H1为普通单倍型,H2为祖先单倍型。(2)草果居群遗传多样性远小于拟草果居群,遗传变异主要来源于居群内,拟草果居群主要来源于居群间。麻栗坡铁厂(TC)、屏边玉屏(YP)居群的遗传多样性、单倍型多样性高于其他18个草果居群。(3)进一步分析表明,包含屏边、马关、西畴、麻栗坡的云南东南部前端地域和邻接的广西那坡可能共同构成草果栽培驯化起源中心,以麻栗坡为核心区域,向周边的西畴、马关、屏边、那坡扩张。因此,结果显示应对TC、YP、那坡下华(XH)居群加以保护。该研究结果为草果种质资源保护、利用提供了遗传学信息。     

Chloroplast DNA diversity associated with protected slopes and valleys for hybridizing Eucalyptus species on isolated ranges in south‐eastern Australia

Aim To relate genetic diversity to topographic features and to investigate genetic interactions between Eucalyptus species in a local centre of endemism and diversity in south‐eastern Australia. Location Grampian Ranges, Victoria, Australia. Methods We documented chloroplast DNA (cpDNA) variation for a group of endemic Eucalyptus species (E. serraensis, E. verrucata and E. victoriana) that dominate rocky, high‐elevation ridgelines of the Grampian Ranges and for one closely‐related, widespread species (E. baxteri) occupying flanking slopes and valleys. We documented genetic patterns across the landscape using cpDNA microsatellites, and related them to topographic features (exposed west‐facing versus protected east‐facing slopes and valleys). We also determined the extent of local haplotype sharing between populations of endemic species and neighbouring E. baxteri downslope with cpDNA microsatellites, and haplotype sharing between the endemic group and more distantly related species (E. obliqua, E. pauciflora and E. willisii) with sequences of the J_LA+ chloroplast region. Results We detected 26 cpDNA microsatellite haplotypes in a relatively small area of c. 20 km × 50 km. Populations of E. baxteri on east‐facing slopes and valleys had greater cpDNA microsatellite diversity than E. baxteri and endemic species on exposed west‐facing slopes. Endemic species frequently shared chloroplast haplotypes with E. baxteri downslope. Sharing of J_LA+ haplotypes with species outside the endemic group was mostly restricted to E. victoriana, which had cpDNA more similar to the species from other sections of Eucalyptus (E. obliqua, E. willisii and E. pauciflora). Main conclusions Intensive sampling of related species on small isolated mountain ranges allowed us to relate genetic diversity to fine‐scale habitats and to document extensive local haplotype sharing between species. This study contributes to a general understanding of the environmental conditions that enable plant population persistence by linking concentrations of genetic diversity to particular habitats.     

Extensive Mitochondrial Introgression from Pinus pumila to P. parviflora var. pentaphylla (Pinaceae)

Pinus species exhibit a paternal chloroplast inheritance and a maternal mitochondrial inheritance. The levels and patterns of cpDNA and mtDNA introgression between the two pine species, P. pumila and P. parviflora var. pentaphylla, were examined at three mountain sites in Japan. The pine species were examined by using PCR-based diagnostic genetic markers of cpDNA and mtDNA. The survey which was carried out in multiple hybrid zones demonstrated a generality in the uni-directional pattern of cytoplasmic gene flow between the two pine species, i.e. paternal cpDNA flowed from P. parviflora var. pentaphylla to P. pumila, and in contrast, maternal mtDNA flowed from P. pumila to P. parviflora var. pentaphylla. Whenever plants which had a non-native combination of cpDNA and mtDNA were observed, they always had the cpDNA haplotype of P. parviflora var. pentaphylla and the mtDNA haplotype of P. pumila. The existence of only this type of cytoplasmic chimera may suggest that F₁ hybrids are successfully produced only in the crossing of P. pumila as the maternal parent and P. parviflora var. pentaphylla as the paternal parent. The present study also detected extensive mtDNA capture in populations of P. parviflora var. pentaphylla located in the southern and middle parts of the Ohu Mountains, Tohoku, Japan. In that area, nearly all of the plants examined had the mtDNA haplotype of P. pumila. The extensive mtDNA introgression suggests that seed flow could be an effective medium for interspecific gene exchange. Received 17 August 1998/ Accepted in revised form 7 January 1999     

祁连山区特有物种黑虎耳草的居群遗传多样性研究

利用叶绿体基因(rbcL和trnS-G)及核糖体DNA内转录间隔区(ITS),对祁连山区狭域分布的特有物种黑虎耳草8个居群(115个个体)进行遗传多样性研究,旨在揭示黑虎耳草的居群遗传结构及其历史进化过程。结果表明:(1)所有取样个体共检测到4个cpDNA单倍型和9个ITS单倍型,其中祁连山东南部的居群固定较多的单倍型和特有单倍型,而西北部居群只固定少数几个广泛分布的单倍型,且遗传多样性普遍较低。(2)基于cpDNA数据和ITS数据的分子变异分析(AMOVA)显示,遗传变异主要来源于居群内。(3)基于cpDNA数据的中性检验表明,Tajima’s D(-1.012 30,P> 0.05)和Fu&Li’s D*(-2.066 77,P> 0.05)均为负值,均不显著;歧点分布分析结果显示,黑虎耳草居群经历过近期的扩张事件。根据物种现有遗传分布格局推测,黑虎耳草在第四纪冰期时退缩到祁连山东南部的边缘避难所,间冰期或冰期后回迁到祁连山西北部地区,在回迁过程中由于奠基者效应导致祁连山西北部的居群仅固定少数广泛分布的单倍型,并呈现出较低的遗传多样性;由于居群较小且相互隔离,该...     

Comparison of genetic population structure between two cyprinids, <Emphasis Type="Italic">Hemigrammocypris rasborella</Emphasis> and <Emphasis Type="Italic">Pseudorasbora pumila</Emphasis> subsp., in the Ise Bay basin,central Honshu,Japan

Two small cyprinid fishes, Hemigrammocypris rasborella and Pseudorasbora pumila subsp. (sensu Nakamura 1963), inhabit similar habitats and often occur sympatrically in the Ise Bay basin, central Honshu Island, Japan. Their genetic population structures were revealed, using sequence data from the mitochondrial cytochrome b gene, and then compared. Hemigrammocypris rasborella populations in the Ise Bay area formed a monophyletic group that has been isolated from eastern (Tenryu River) and western (Lake Biwa–Yodo River) populations at least for several hundred thousand years. Pseudorasbora pumila subsp., endemic to the Ise Bay area, was estimated to have become isolated from its sister subspecies, P. p. pumila, about 5 million years ago. Both H. rasborella and P. pumila subsp. had centers of genetic diversity around the Okazaki Plain in the eastern part of the basin and showed trans-bay distribution of haplotypes or haplotype groups. Their common population structure was explained by geological features in the Ise Bay area, in which a large paleo-river system developed in regression periods, suggesting gene flow among populations of each species in the mid to lower reaches of the paleo-river. Based on the estimated expansion or divergence time, however, not all populations experienced gene flow during the Last Glacial. In contrast to the maintenance of high genetic diversity in H. rasborella, almost all populations of P. pumila subsp. have lost mitochondrial DNA genetic diversity. This implies that effective population size of P. pumila subsp. tended to be smaller, probably because of differences in reproductive ecology, even though the two species have been exposed to similar environmental changes. For conservation of the two species, genetic and adaptive differentiation among local populations should be considered, and attention should be paid to inbreeding depression, especially in P. pumila subsp. An erratum to this article can be found at     

Population structure of <Emphasis Type="Italic">Nouelia insignis</Emphasis> (Asteraceae), an endangered species in southwestern China,based on chloroplast DNA sequences: recent demographic shrinking

Nouelia insignis, an endangered species, is distributed in the Jinsha and Nanpan drainage areas in southwestern China. In this study, we examined the genetic diversity and population structure based on the sequences of the cpDNA rpL 16 intron. Low levels of genetic variation were detected within all populations of the endemic species. A gene genealogy of 11 haplotypes recovered two major lineages I and II, with haplotypes H1 and H6 nested as interior nodes, respectively. Haplotype H1 was widespread in all populations, while haplotype H6 was restricted to populations southern of the Jinsha River. Low levels of genetic differentiation were detected, as most F _st values between populations were zero. This result, however, contradicts previous studies based on allozymes and fingerprinting. Genetic analyses suggested that coancestry due to low evolutionary rates resulted in the lack of geographical subdivision. Molecular dating estimated that the two lineages split about 3.224 MYA (95% CI 1.070–6.089 MYA). Maintenance of ancestral polymorphisms was possibly attributable to a long-standing large effective population size until recently. Postglacial demographic expansion was supported by a unimodal mismatch distribution and star-like phylogenies.     

Protecting evolutionary significant units for the remnant populations of <Emphasis Type="Italic">Berchemiella wilsonii</Emphasis> var. <Emphasis Type="Italic">pubipetiolata</Emphasis> (Rhamnaceae)

Berchemiella wilsonii var. pubipetiolata (Rhamnaceae) is an endangered tree in eastern China. Habitat destruction has resulted in fragmentation of remnant populations and extinction of local populations. AFLP and cpDNA markers were used to determine the population structure of remnant populations of B. wilsonii var. pubipetiolata. Moderate nuclear genomic diversity was found within each of the four remnant populations (H _S = 0.141–0.172), while the cpDNA haplotype diversity in each population ranged from 0.356 to 0.681. Six haplotypes were identified by a combined cpRFLP and cpSSR analysis in a total of 89 individuals. AMOVA revealed significantly AFLP genetic differentiation within and between regions (Φ_SC = 0.196, Φ_CT = 0.396, respectively), and a high cpDNA haplotype differentiation between regions (Φ_CT = 0.849). The results suggest low gene flow between populations of B. wilsonii var. pubipetiolata. Strong genetic divergence between two regional populations as revealed by both AFLP and cpDNA markers provided convincing evidence that two distinct evolutionary lineages existed, and should be recognized as ‘evolutionary significant units’ (ESUs) for conservation concerns.     

Limited structure and widespread diversity suggest potential buffers to genetic erosion in a threatened grassland shrub Pimelea spinescens (Thymelaeaceae)

Pimelea spinescens is a critically endangered species of the temperate grasslands of southeastern Australia. Two subspecies are recognised. Subspecies, P. spinescens subsp. spinescens, formerly common and widespread, is found in isolated remnants of previously extensive grasslands. The second subspecies, subsp. pubiflora, is thought to have been historically rare with only two geographically-isolated extant populations. The grassland communities exist now as fragmented remnants representing <1 % of their extent prior to European settlement in the early 1800s. Conservation management strategies for species in these critically endangered ecosystems rely on an understanding of genetic diversity and population structure to ensure long-term evolutionary potential. We used chloroplast DNA (cpDNA) and microsatellite markers to examine the population genetic structure of both subspecies. Analysis of cpDNA revealed 14 haplotypes with high divergence between the single haplotype found in subsp. pubiflora and most remaining haplotypes restricted to subsp. spinescens. Microsatellites also indicated high genetic differentiation between subspecies but little evidence of sub-structuring within either subspecies. Results suggest that seed dispersal has not been as limited as previously thought. In this fragmented habitat, a lack of genetic structure suggests buffers to genetic erosion, with current patterns reflecting genetic diversity prior to fragmentation. Plant longevity and the presence of seed banks may contribute to the maintenance of these patterns, resulting in a lag between fragmentation and genetic erosion. Whilst factors such as longevity and seed banks may be preserving historic genetic diversity, management is required to ensure the maintenance of this diversity into the future.     

Genetic variation in populations of the morphologically and ecologically variable fern <Emphasis Type="Italic">Stegnogramma pozoi</Emphasis> subsp. <Emphasis Type="Italic">mollissima</Emphasis> (Thelypteridaceae) in Japan

In Japanese Stegnogramma pozoi subsp. mollissima (Fisher ex Kunze) K. Iwats. there is the intrasubspecific variation among rbcL sequences. Northern and southern plants are genetically differentiated for maternally inherited cpDNA. In the present study we examined allozyme polymorphisms to test the hypothesis that northern and southern plants may be separate species. Based on allozyme data, the degree of gene flow among populations was estimated to be large. The artificial crossing experiments between cpDNA haplotypes also suggested that isolation has not developed among these cpDNA haplotypes. However, interpopulation genetic differentiation in cpDNA was observed even in the small area at the foot of Mt. Hakone, and the cpDNA haplotypes appear to have different habitat preferences. Received: May 1, 2001 / Accepted: September 30, 2001