Genetic Diversity Within and Among Populations of Viola tenuicornis with Reference to Sampling Strategies

Genetic diversity and population differentiation of Viola tenuicornis W. Beck. were studied in nine populations in Beijing using horizontal starch gel electrophoresis. Allozyme data for 11 loci of 8 enzymes indicated high level of genetic variability in this region with P=72.7％, A=2.4 and H e=0.243. In contrast, the population differentiation was lower than the average of other perennial herb with F st=0.196. It was found that distances among populations had little effect on population differentiation. A sample of 30 individuals could well represent a population, but less than 20 individuals per population might lead to significant bias in the estimation of genetic parameters, in particular the values of A and H e.     

东北地区野豌豆属Vicia L.物种生物学研究 Ⅲ.广布野豌豆类群细胞学初步观察

本文首次报道了国产二倍体广布野豌豆的核型, 其核型公式分别为2n=12=2m+8sm(2SAT)+2st, 2n=14=2m+8sm(2SAT)+4st。其中2n=14的染色体计数系我国新纪录。为修订国产广布野豌豆的分类处理提供了必要的细胞学依据。     

A Further Study of the Genus Viola in Hebei Province

In this study, 23 OTU’s including all the species of Viola in Hebei Province were used in numerical taxonomic treatment. According to the principle of clustering line method, a few lines which divide sections, subsections and species were respectively decided. The result of numerical research roughly coincides with the scheme of 3 sections and 4 subsections proposed by Gingins et al.. Based on the value of distance coefficient, morphological characters and geographical distribution, the species V. pekinensis is reduced as a synonym of V. mongolica. The writer compared section Melanium with other sections with characters of morphology, palynology, cytology, phytogeography as well as pollination ecology and sugges ted a revision of systematic position of section Melanium.     

Morphological versus genetic diversity of Viola reichenbachiana and V. riviniana (sect. Viola,Violaceae) from soils differing in heavy metal content

Morphological characters, AFLP markers and flow cytometry were used to investigate the morphological and genetic variability and differentiation of Viola reichenbachiana and V. riviniana in non‐metallicolous (NM) and metallicolous (M) populations. The aims were to clarify the taxonomic status of plants occurring in ore‐bearing areas, to determine any relationship in V. reichenbachiana and V. riviniana from sites not polluted with heavy metals, and to examine the genetic variability and differentiation of M and NM populations of both species. Multivariate analyses based on morphological characters showed significant differences between V. reichenbachiana and V. riviniana from non‐polluted sites, high levels of intra‐ and inter‐population variability, and the occurrence of inter‐specific hybrids. Plants from M populations showed hybrid characters but also fell within the range of V. riviniana or V. reichenbachiana. There were no significant differences in relative genome size between plants from polluted areas and V. riviniana from NM populations. Bayesian analysis of population genetic structure based on AFLP markers distinguished two main groups: V. reichenbachiana and V. riviniana together with the M populations. That analysis also revealed the occurrence of populations of inter‐specific hybrids from non‐polluted areas. Further Bayesian analysis of V. riviniana including NM and M populations separated all the studied M populations from NM populations. We conclude that plants forming the M populations are well adapted to a metal‐polluted environment, and could be considered as stabilised introgressive forms resulting from unidirectional (asymmetric) introgression toward V. riviniana.     

Soil diaspore reserves above the timberline in the Austrian Alps

Soil diaspore reserves are considered to support self-healing processes after vegetation disturbances. Therefore, the stratified reserves of viable diaspores in superimposed soil layers of four sites above the timberline in the Austrian Alps were assessed. At each site, a semi-natural (“undisturbed”) extensive alpine pasture and the disturbed vegetation on hiking trails were investigated. Eighty soil cores in total (corresponding to 400 slices, each representing a 1-cm layer between 0 and 5 cm depth) were taken in autumn and subjected to germination tests after vernalization. The total diaspore numbers in disturbed and undisturbed plots did not differ significantly, but all undisturbed soils contained higher species numbers than disturbed ones. Seed shape and size clearly influenced the vertical distribution. Intact soils showed a significant decrease in big/long diaspores with increasing soil depth. Disturbances influenced the aboveground species composition and therefore the distribution of seeds of different size. In case of disturbances, the restriction of most big seeds to superficial layers means a threat for small populations of rare and protected species such as Viola lutea subsp. sudetica with relatively big seeds near the soil surface. When the disturbances stop, the diaspore communities might initiate a first, but with respect of landscape protection and preservation of species diversity insufficient step of vegetation restoration.     

广东南岭自然保护区堇菜属植物垂直分布格局的研究

在南岭自然保护区核心区选择海拔高度为500—600m、600—800m、800—1000m、1000—1200m、1200—1400m、1400—1600m、1600—1800m、1800—1902m共8个连续的海拔梯度,分别对植物群落中的堇菜属植物进行普查。结果表明,随着海拔升高,各海拔段有堇菜属植物5、8、9、10、9、7、3、1种,以中海拔段的物种多样性较高。两海拔段间距离越近,物种相似性系数越大;距离越远,相似性系数越小。在中海拔段,堇菜属植物种类的多度最大,两端较小。南岭自然保护区堇菜属植物的垂直分布格局呈“中间膨胀型”或“单峰型”,与普遍的木本植物一致,这种格局可能是与沿海拔水热梯度的变化相关。     

Genetic and morphological variation in Viola suavis s.l. (Violaceae) in the western Balkan Peninsula: two endemic subspecies revealed

The Balkan Peninsula, with many endemic species, is known as one of the most important speciation and diversification centres in Europe. Here, we present a study of the western Balkan populations of the polymorphic European species, V. suavis s.l., which have been reported under the name V. adriatica, but their taxonomic status and position within the genus have remained uncertain. Viola suavis s.l. and nine close relatives sampled across Europe were subjected to molecular (sequencing of nuclear ribosomal internal transcribed spacers and amplified fragment length polymorphism), karyological and morphometric analyses. Our results revealed the presence of four allopatric, genetically and morphologically differentiated lineages within V. suavis s.l. in Europe, which are suggested here to be recognized at the subspecific rank. Populations from the western Balkans were segregated into two distinct entities: (1) those from north-western Croatia correspond to the previously recognized taxon, V. suavis subsp. adriatica and (2) those from southern Dalmatia (southern Croatia, southern Bosnia and Herzegovina, and south-western Montenegro) are described here as V. suavis subsp. austrodalmatica subsp. nov. The other two lineages of V. suavis s.l., which both harbour blue- and white-flowered morphotypes, occur in central and eastern Europe (V. suavis subsp. suavis) and in north-eastern Spain (plants provisionally treated as V. suavis ‘Spain’). The AFLP and morphological data indicate gene flow between the nominate subspecies and V. suavis subsp. adriatica in a few localities. The distribution of the two western Balkan subspecies is discussed and an identification key to the V. suavis subspecies in Europe is presented.     

Molecular markers in Viola L. subsect. Viola: Application and taxonomic implications for the identification of dubious herbarium specimens

Abstract

The authors report the use of nuclear [internal transcribed spacer (ITS) sequences 1 and 2] and chloroplast DNA (trnS-trnG intergenic spacer sequences) in Viola subsect. Viola for separate tracking of maternal lineages and detecting dubious herbarium specimens. A phylogenetic investigation carried out on ITS data after removal of all material with possible hybrid origin showed that V. hirta is a monophyletic unit, whereas V. odorata includes at least V. collina and V. jaubertiana, as well as three sequences of V. alba subsp. dehnhardtii from literature. In some, among the sampled individuals, the morphological attribution to one species is contradicted by nuclear DNA, which indicated a wide distance from other non-specific individuals from different locations and closer proximity to a different species. Chloroplast DNA data for the same individuals, on the contrary concurred with morphological evidence. These findings confirm document univocal correlation between our chosen chloroplast sequences and the studied taxa at species or subspecies level; these sequences have the appropriate variability range to be employed for detection of the maternal lineage of unknown Viola samples.     

Metabolomics for the Authentication of Natural Extracts Used in Flavors and Fragrances: the Case Study of Violet Leaf Absolutes from Viola odorata

Natural extracts used in fine fragrances (alcoholic perfumes) are rare and precious. As such, they represent an interesting target for fraudulent practices called adulterations. Absolutes, important materials used in the creation of perfumes, are obtained by organic solvent extraction of raw plant materials. Because the nonvolatile part of these natural extracts is not normalized and scarcely reported, highlighting potential adulterations present in this fraction appears highly challenging. For the first time, we investigated the use of nontargeted UHPLC‐ToFMS metabolomics for this purpose, considering Viola odorata l ., a plant largely used in the perfume industry, as a model. Significant differences in the metabolic fingerprints of the violet leaf absolutes were evidenced according to geographical locations, and/or adulterations. Additionally, markers of the geographical origin were detected through their molecular weight/most probable molecular formula and retention time, while adulterations were statistically validated. In this study, we thus clearly demonstrated the efficiency of UHPLC‐ToFMS‐based metabolomics in accelerating both the identification of the origin of raw materials as well as the search for potential adulterations in absolutes, natural products of high added value.     

Microwave-assisted extraction of cyclotides from Viola ignobilis

Cyclotides are an interesting family of circular plant peptides. Their unique three-dimensional structure, comprising a head-to-tail circular backbone chain and three disulfide bonds, confers them stability against thermal, chemical, and enzymatic degradation. Their unique stability under extreme conditions creates an idea about the possibility of using harsh extraction methods such as microwave-assisted extraction (MAE) without affecting their structures. MAE has been introduced as a potent extraction method for extraction of natural compounds, but it is seldom used for peptide and protein extraction. In this work, microwave irradiation was applied to the extraction of cyclotides. The procedure was performed in various steps using a microwave instrument under different conditions. High-performance liquid chromatography (HPLC) and matrix-assisted laser desorption/ionization time-of-flight (MALDI–TOF) results show stability of cyclotide structures on microwave radiation. The influential parameters, including time, temperature, and the ratio of solvents that are affecting the MAE potency, were optimized. Optimal conditions were obtained at 20 min of irradiation time, 1200 W of system power in 60 °C, and methanol/water at the ratio of 90:10 (v/v) as solvent. The comparison of MAE results with maceration extraction shows that there are similarities between cyclotide sequences and extraction yields.