落叶松种间及无性系间ISSR鉴别技术的研究

采用ISSR分子标记技术对兴安、长白和日本落叶松种间以及不同无性系进行了鉴别。从49条引物中筛选出13条ISSR引物可以对落叶松种间和无性系间进行鉴别,特异条带个体的百分率为100%,该项技术为落叶松新品种以及良种的准确鉴别提供了新的途径和手段：其中5条引物在日本落叶松、兴安落叶松和长白落叶松不同位置扩增出特异谱带,作为种的鉴定的标准,有9条引物可以对落叶松种内不同无性系分别扩增出特异片段,进行无性系的鉴别。     

Larch chromosomal mosaicism

Karyological study of larch population members from different parts of the Larix olgensis L. Henry areal in the Primorskii krai has been carried out. The main amount of chromosomes for larch as n = 12 (2n = 24) has been confirmed. Mixoploidy was observed in all studied populations. The difference of individuals from the larch areal based on the cell amount with a different level of ploidity has been found in the population of L. olgensis locus classicus and both L. sibirica and L. gmelinii, which is probably a consequence of their hybrid nature.     

落叶松幼苗过氧化氢酶动力学的初步研究

采用碘量法对4种落叶松幼苗的过氧化氢酶（CAT）动力学进行了研究。结果表明：在25℃、pH 7.0下,落叶松幼苗CAT活性为48.8～77.9 U·g^-1 FW;Km值为8.3×10-2～2.38×10^-1 mol·L^-1;Vmax为0.071～0.200 μmol·min^-1;CAT最适温度15～25℃,对温度的适应范围华北落叶松和兴安落叶松宽,长白落叶松和太白落叶松的窄;CAT最适pH值为7.0。太白落叶松对pH适应范围最窄,其可能是退化的物种。     

Seed-specific identification of Larix gmelinii,Larix olgensis,and Larix principis-rupprechtii using sequence-characterised amplified region markers

Larix gmelinii, Larix olgensis, and Larix principis-rupprechtii are the three native and sympatric larch species in North China, and each of these species has a distinctive ecological niche. It is difficult to identify them based only on certain morphological characters, particularly the seed appearance. In this study, the seed endosperms of these three larch species were analysed using the random amplified polymorphic DNA (RAPD) technique to screen for interspecific differences. The following three RAPD markers linked to species-specific segments were observed in the different species: 1475-bp (Larix gmelinii and L. olgensis), 505-bp (Larix principis-rupprechtii), and 1121-bp (Larix gmelinii) markers. The three seed-specific fragments amplified by the RAPD markers were sequenced, and the sequences were used to design and synthesise species-specific SCAR markers. The size of the SCAR fragments was concordant with that of the RAPD species-specific fragments. Therefore, these SCAR markers can be used to identify the seeds of different larch species, thereby providing a new molecular tool for the identification of larch seeds that leads to considerable savings in terms of time and economic resources.     

Genetic variability and differentiation in the larch populations within the range of <Emphasis Type="Italic">Larix olgensis</Emphasis> A. Henry in Primorye

The level of within-and among-population variation of larch inhabiting the range of Larix olgensis A. Henry in Primorye was estimated on the basis of 440 RAPD loci identified by means of 12 random primers. In ten populations examined, the proportion of polymorphic loci was 35–60%, the average expected heterozygosity varied from 0.1340 to 0.2169, and the average gene flow estimate was 1.38. According to Fisher’s test for heterogeneity, the pairwise differences of the fragment frequencies between the populations were statistically significant. The subdivision index G _ST = 0.2663 indicated that the interpopulation variation component accounted for approximately 27% of the total variation. Coefficients of Nei’s genetic distance between the populations varied from 0.0137 to 0.0934. Most of the samples with high support clustered according to the geographic positions relative to one another within the range. These results suggest that the populations examined are characterized by high genetic variation, like the larch populations of Siberia and the Russian Far East studied earlier, but, in contrast to the latter, exhibit higher among-population variability.     

Morphometric Differences between Two Smelt Species, <Emphasis Type="Italic">Hypomesus japonicus</Emphasis> (Brevoort, 1856) and <Emphasis Type="Italic">H. nipponensis</Emphasis> (McAllister, 1963) (Pisces: Osmeridae) from The Northwestern Part of the Sea of Japan

Two smelt species that inhabit the northwest of the Sea of Japan, Hypomesus japonicus and H. nipponensis (family Osmeridae), which are both currently referred to as Japanese smelt, have been studied in order to clarify the taxonomic value of morphometric characters. Variations between individuals of these species have been revealed using a multivariate analysis of their morphometric characters and indices. The results of the analysis show that H. japonicus is distinguished from H. nipponensis both by the previously known characters (the eye diameter is smaller and the vertical through the beginning of the dorsal fin is closer to the head than that through the beginning of the pelvic fins) and by a greater height of the dorsal and adipose fins in the former species as defined in the paper. Within the species H. japonicus, the individuals sampled from Olga Bay are most similar morphologically to those from the coastal waters off the village of Terney. In contrast, the individuals of this species from the coastal waters off Russky Island and from Olga Bay manifest the greatest differences.     

Investigation of physicochemical properties of arabinogalactan of different larch species

Samples of arabinogalactan (AG) were isolated by aqueous extraction from wood of different larch species (Larix sibirica Ledeb., Larix gmelinii (Rupr.) Rupr., Larix cajanderi Mayr., and Larix olgensis var. Koreana) and studied by HPLC, IR spectroscopy, and quantitative ¹³C NMR spectroscopy. The wood of the studied larch species was shown to contain significant amount of arabinogalactan (10–17% of the a.d.w. mass) and could be a source of its industrial production. The studied AG samples had similar structural and molecular-mass characteristics. This fact could facilitate the product standardization during its industrial production both from the wood of a single larch species and from mixed raw materials.     

Genetic diversity of larch at the north of Primorskii Krai and limits of Larix olgensis A. Henry distribution

The genetic diversity of four mtDNA fragments and five microsatellite loci of cpDNA was examined in six larch population samples from the territory of the Sikhote-Alin Nature Reserve (northern Primor’e). In order to identify possible species-specific differences between the samples, the latter were collected at transects along the shore of the Sea of Japan and at different distances from the sea. Based on a number of morphological characters, some authors suggest that Olgan larch (Larix olgensis A. Henry) grows in the shoreline part of the reserve and, moving inland, it is replaced by Gmelin larch (Larix gmelinii (Rupr.) Rupr.). According to the other data, the northern border of the Olgan larch range does not reach the reserve territory. The data obtained were compared to those obtained previously for three samples from the south of Primor’e, including those for the locus classicus of Olgan larch. In the examined larch individuals (186, for mtDNA and 200, for cpDNA), a total of five mitotypes and 52 chlorotypes were recovered. According to the results of the AMOVA test, the proportion of variations accounted for the differences among all Larix population samples examined over both types of markers was higher (N _ST = 0.435, for mtDNA and R _ST = 0.041, for cpDNA) than that for the differences among the population samples within the reserve (N _ST = 0.079, for mtDNA and R _ST = 0.005, for cpDNA). No differences were detected between the groups of shoreline and continental populations.     

MeJA is more effective than JA in inducing defense responses in <Emphasis Type="Italic">Larix olgensis</Emphasis>

The roles of jasmonic acid (JA) and methyl jasmonate (MeJA) in improving the inducible resistance of plants to biotic and abiotic stimuli/stresses have been well investigated. However, the differences in inducing effects between exogenous applications of JA and MeJA are poorly understood. In this study, we compared the inducing effects of exogenous spray applications of 0.1 mmol/L JA and MeJA onto four un-bagged lateral branches on defense response of Larix olgensis seedlings against the gypsy moth (Lymantria dispar). The bio-activities of three major defense enzymes (SOD, PAL, and PPO) plus two protease inhibitors (TI and CI) of the unsprayed larch seedling needles, and the growth, development and reproductive capacity of the gypsy moth were examined. The results show that partial spray of JA or MeJA on L. olgensis seedlings significantly increased the bio-activities of SOD, PAL, PPO, TI, and CI (P < 0.05), and strongly decreased the larval/pupal weights and survivals, as well as the fecundity of L. dispar that fed on the seedlings relative to the control. However, the MeJA treatment showed quicker inductive effects on SOD and PAL activities; longer and more significant effects on PPO, TI, and CI activities; better inhibitory effects on the larval/pupal weights and survivals, as well as the fecundity of L. dispar than did the JA treatment. Comparatively, MeJA in the current study showed stronger effects on inducing systemic resistance to the defoliator (L. dispar) in L. olgensis than did JA.     

Horizontal gene transfer from a flowering plant to the insular pine Pinus canariensis (Chr. Sm. Ex DC in Buch)

Horizontal gene transfer (HGT) is viewed as very common in the plant mitochondrial (mt) genome, but, to date, only one case of HGT has been found in gymnosperms. Here we report a new case of HGT, in which a mt nad5-1 fragment was transferred from an angiosperm to Pinus canariensis. Quantitative assay and sequence analyses showed that the foreign nad5-1 is located in the mt genome of P. canariensis and is nonfunctional. An extensive survey in the genus Pinus revealed that the angiosperm-derived nad5-1 is restricted to P. canariensis and present across the species'' range. Molecular dating based on chloroplast DNA suggested that the HGT event occurred in the late Miocene after P. canariensis split from its closest relatives, and that the foreign copy became fixed in P. canariensis owing to drift during its colonization of the Canary Islands. The mechanism of this HGT is unclear but it was probably achieved through either direct cell–cell contact or external vectors. Our discovery provides evidence for an important role of HGT in plant mt genome evolution.     

Pliocene wood of Larix from southern Primorye (Russian Far East)

A new fossil larch species, Laricioxylon blokhinae, showing the wood anatomy of modern Larix olgensis A. Henry and L. leptolepis (Siebold et Succ.) Gord. is described. The taxonomic and structural diversity of larch species is reviewed, based on fossil wood remains from the Pliocene of southern Primorye.     

Development of a sequence characterized amplified region (SCAR) marker associated with high rooting ability in <Emphasis Type="Italic">Larix</Emphasis>

In this study, bulked segregant analysis (BSA) was used on Larix leptolepis × Larix olgensis hybrids to identify a random amplified polymorphic DNA (RAPD) marker associated with high rooting ability in larch. Two DNA bulks: H (high rooting ability) bulk and L (low rooting ability) bulk were constructed according to the rooting percentages of the stock plants. Among the 328 primers, only S356 could amplify a specific band, named S356₄₄₅, which only existed in the H bulk and was further confirmed following selective genotyping of individual hybrids. Grounded on the border sequences, S356₄₄₅ was converted to a sequence characterized amplified region (SCAR) marker, HRL445, which can be useful in marker-assisted selection (MAS) to screen for larch with high rooting ability. All the results strongly indicated that S356₄₄₅ and HRL445 were closely associated with high rooting ability in larch.     

Disturbance and regeneration dynamics of a mixed Korean pine dominated forest on Changbai Mountain,North-Eastern China

We used dendrochronological methods to study disturbance history of a mixed Korean pine (Pinus koraiensis Siebold et Zuccarini) dominated forest on the northern slope of Changbai Mountain, North Eastern China, over 1770–2000. Frequent small-scale canopy gaps and infrequent medium-scale canopy disturbances dominated natural disturbance regime in the forest, which did not experience stand-replacing disturbances over the studied period. Percentages of growth releases in subcanopy trees were below 6% in most decades, suggesting that disturbances initiating these releases were of low intensity. Strong winds were likely cause of moderate disturbance events. Two episodes with increased disturbance rates (19% and 13%) were dated to the 1920s and 1980s, timing of the 1980s event was consistent with a hurricane occurred in 1986 on the western slope of the Changbai Mountain. Age structure and growth release analyses revealed species-specific regeneration strategies of canopy dominants. Shade-intolerant Olga bay larch (Larix olgensis Henry) recruited mainly before the 1860s. Recruitment of moderately shade-tolerant P. koraiensis occurred as several regeneration waves (1820s, 1850s, 1870–1880s, 1930s, and 1990–2000s) of moderate intensity. Shade-tolerant Jezo spruce (Picea jezoensis Carr. var. komarovii (V. Vassil.) Cheng et L.K.Fu) and Manchurian fir (Abies nephrolepis (Trautv.) Maxim.) regenerated continuously over the last 220 and 130 years, respectively. Enhanced recruitment of P. koraiensis, P. jezoensis, and A. nephrolepis was observed during the 1930s and 1990s, coinciding with increased growth release frequency in the 1920s and 1980s, and suggesting disturbance events of moderate intensity. Our results indicate that the current disturbance regime of the mixed Korean pine dominated forest maintains coexistence of light-demanding and shade-tolerant species and that change in wind climate may be particularly important for future forest composition.     

Genotyping Echinococcus granulosus from dogs from Western Iran

Cystic echinococcosis is a zoonotic infection caused by the dog tapeworm, Echinococcus granulosus. In the present study, adults of E. granulosus (n = 20) were collected from 71 dogs from Western Iran and were genetically characterized using DNA sequencing of the partial mitochondrial cytochrome c oxidase subunit 1 (cox1) and NADH dehydrogenase 1 (nad1). Consensus sequences were obtained for cox1 (366) and nad1 (471) genes. Phylogenetic analysis of concatenated nad1 and cox1 nucleotide sequence data was performed using Bayesian Inference approach. Overall, the dog isolates indicated nine different sequences in cox1 and seven in nad1 genes. Three genotypes (G1 [75%], G2 [10%] and G3 [15%]) were identified from the isolates. The G2 sequences indicated 100% homology with reference G2 sequence in both cox1 (Genbank accession number M84662) and nad1 (AJ237633) genes. G3 sequences showed 100% homology with G3 reference sequence in nad1 (AJ237633), but displayed two different cox1 profiles, each having 99% homology with reference G3 sequence (M84663). In the phylogenetic tree all of the isolates were grouped into a distinct cluster corresponding to the G1–G3 complex with relevant reference sequences. The presence of G1 genotype (sheep strain) of E. granulosus sensu stricto as dominant genotype in dogs is emphasized. To the best of our knowledge, this study established the first record of E. granulosus sensu stricto, G2 genotype in Iran.     

落叶松人工林生物量模型研究

以不同年龄、不同密度的落叶松（Larix olgensis）人工林为研究对象,基于19块标准地95株标准木的树干解析、枝解析的生物量数据,研究不同大小树木因子（胸径、树高、冠幅等）与单木各分量（树干、枝、叶）生物量之间的关系,应用统计分析软件建立落叶松单木各部分生物量的回归模型。利用单木各部分生物量回归模型方程估测落叶松人工林各林分的总生物量,并分析了不同年龄及林分密度下林分生物量的变化规律：林分的生物量随年龄的增加而不断增长,树干的生物量的比例是最大的,同时也是随着年龄的增长而不断的增加,而树枝和树叶的生物量的比例比较小,林分的生物量随林分密度的增加而不断增加。最后建立林分生物量模型,为落叶松人工林的研究提供基础资料,为了解落叶松人工林的生产力,对其进行合理经营提供科学依据。     

New record of Ascaridia nymphii (Secernentea: Ascaridiidae) from macaw parrot,Ara chloroptera,in China

Present study was performed to identify the species of ascarids from macaw parrot, Ara chloroptera, in China. Total 6 ascarids (3 males and 3 females) were collected in the feces of 3 macaws at Guangzhou Zoo in Guangdong Province, China. Their morphological characteristics with dimensions were observed under a light microscope, and their genetic characters were analyzed with the partial 18S rDNA, ITS rDNA and nad4 gene sequences, respectively. Results showed that all worms have no interlabia but male worms have two alate spicules, well-developed precloacal sucker and a tail with ventrolateral caudal alae and 11 pairs of papillae. The partial 18S rDNA, ITS rDNA and nad4 sequences were 831 bp, 1015 bp and 394 bp in length, respectively. They showed the highest similarity of 99.8% (18S rDNA) with Ascaridia nymphii, 93.8% identities (ITS rDNA) with A. columbae and 98.5% to 99.5% identities (nad4) with Ascaridia sp. from infected parrot. All Ascaridia nematodes from the macaws were clustered into one clade and formed monophyletic group of Ascaridia with A. columbae and A. galli in two phylogenetic trees. It is observed that the combining morphological and sequencing data from three loci, the present Ascaridia species was identified as Ascaridia nymphii, which is the first record of A. nymphii from macaw parrot in China.     

Exploring genetic variations in threatened medicinal orchids using start codon targeted (SCoT) polymorphism and marker-association with seed morphometric traits

We aimed to study the genetic diversity and population structure of eight Iranian terrestrial orchid species, including Anacamptis coriophora (L.) R. M. Bateman, Pridgeon and M. W. Chase, Dactylorhiza umbrosa (Kar. & Kir.) Nevski, Himantoglossum affine (Boiss.) Schltr., Orchis collina Banks and Solander, Orchis mascula (L.) L., Orchis simia Lam., Ophrys schulzei Bornm. and Fleischm., and Ophrys straussii H. Fleischm. and Bornm. using start target codon markers (SCoT) and finding markers associated with seed morphometric traits. A total of 254 reproducible SCoT fragments were generated, of which 248 fragments were polymorphic (average polymorphism of 96.18%). The SCoT markers showed a narrow range of polymorphism information content (PIC) varied from 0.397 for S9 primer to 0.499 for S11 and S20 primers. Based on the population analysis results, the Orchis simia accessions collected from Paveh region (Os.P) represented the lowest observed number of alleles (Na) (1.13) and effective number of alleles (Ne) (1.09). At the same time, the highest Na (1.29) and Ne (1.18) values were obtained in O. schulzei collected from Javanrood (Oyst.JA). Shannon’s information index (I) was ranged from 0.03 for D. umbrosa accessions collected from Marivan (Du.M population) to 0.263 for Ha.Ja population (H. affine accessions collected from Javanrood). The UPGMA dendrogram obtained with the Jaccard similarity coefficient (r = 0.97295) divided 97 studied terrestrial orchid accessions into eight groups mainly based on species type and geographical origin. Based on the Bayesian statistical index, the highest probability of the data was achieved when accessions were divided into eight groups (K = 8). Multiple association analysis (MRA) revealed significant associations between some of SCoT bands with seed morphometric traits. Our findings can be useful for germplasm characterization, conservation, and improvement of Iranian terrestrial orchid species.Supplementary InformationThe online version contains supplementary material available at 10.1007/s12298-021-00978-4.     

Fifty mosses on five trees: comparing phylogenetic information in three types of non-coding mitochondrial DNA and two chloroplast loci

Given the frequent genomic recombinations in plant mitochondrial DNA, intergenic regions of this organelle genome had so far not been considered as loci of potential phylogenetic information. Based on the recent evidence that an evolutionary ancient mitochondrial nad5-nad4 gene continuum is conserved in bryophytes we have compiled a dataset for a phylogenetically wide sampling of 50 mosses covering this intergenic region. The length of the intergenic region was generally in the range of 585 (Diphyscium) to 646 bp (Tomentypnum) with rare exceptions, for example all Polytrichales taxa showing a 200 bp deletion as an apparent synapomorphy of this order. Phylogenetic information in the novel marker sequence was compared with that of a nad5 gene region containing a group I intron and a nad2 gene region containing a group II intron as well as with two widely sampled chloroplast data sets, rbcL and rps4. Indel evolution in the three types of non-coding mitochondrial sequences is obviously more taxon-dependent than locus-dependent, indicating lineage-specific insertion/deletion rates. For example, larger sequence deletions are a general feature in Schistostega and Tetraphis. Although confidence for particular nodes in the phylogeny was found to vary among data sets, gene trees were essentially without conflict with respect to well-supported ones and add up in information towards a reasonably well-resolved moss phylogeny. However, while a consensus on the latter is clearly emerging, sufficient confidence is still lacking for the first dichotomies among the arthrodontous mosses leading into subclasses Bryidae, Dicranidae and Funariidae and the relative placement of nematodontous mosses (Polytrichales and Tetraphidales) on the backbone of early moss phylogeny.     

An insight into the genetic polymorphism among European populations of Lactuca serriola assessed by AFLP

Prickly lettuce (Lactuca serriola) is world-wide distributed and very variable species generally considered as a progenitor of the cultivated lettuce (Lactuca sativa). Altogether, 50 populations of L. serriola were characterized by means of amplified fragment length polymorphism (AFLP) and by isozyme analysis. Relationships among individuals and populations were examined by applying the unweighted pair-group method with the arithmetic averages (UPGMA) clustering algorithm, principal coordinate analysis (PCA) and the Nei's gene diversity index. The studied set of populations split into three main groups based on the AFLP polymorphism analysis. The first group contained L. sativa (control). The second group comprised two L. serriola accessions; one of them was identified as L. serriola f. integrifolia and the other as a mixture of two L. serriola forms. The largest and the most diverse third group contained the remaining L. serriola accessions. The population clustering corresponded approximately to their geographical distribution in Europe. At least five distinct geographic groups were recognised: 1) Northern European; 2) Slovenian; 3) very heterogeneous Central and Western European (mostly north of the Alps); 4) Mediterranean; 5) prevalence of L. serriola f. integrifolia, mostly comprising accessions from the United Kingdom and the Netherlands. This study showed that accessions originating in various eco-geographical conditions of Europe differ significantly in their genetic and protein polymorphism, as well as in morphology. Some European L. serriola populations (e.g. from Scandinavia and United Kingdom/British Isles/) seems to be isolated and homogeneous; in contrast, populations occurring in Central Europe are very diverse and genetically overlapping.