The complete mitogenome of Anaplectoides virens (Butler, 1878) (Lepidoptera: Noctuidae) and phylogenetic analysis.

Anaplectoides virens (Noctuoidea: Noctuidae) is a polyphagous herbivorous moth, which feeds on a wide variety of crops. Molecular phylogenetic studies of this species are still limited. We presented the first complete mitochondrial genome of the genus Anaplectoides, which was assembled from data generated using a genome skimming method. The assembled mitogenome is 15,358 bp in length and consists of 37 genes, including 13 protein-coding genes, two rRNAs, 22 tRNAs, and a control region. Except for the start codon of cox1 with CGA, other coding genes use ATN as the start codon. Most PCGs use TAA as the stop codon; however, cox1, cox2, and nad4 use T as the termination codon. Phylogenetic analysis revealed that the genera of ((Agrotis + Striacosta) + Anaplectoides) within Noctuinae formed a monophyletic group. Among Noctuidae, the relationship of ((Noctuinae + Hadninae) + Amphipyrinae) was also highly supported.

On the identity of Conisania leuconephra Draudt, 1950 with a description of a new species from Xizang, China (Lepidoptera: Noctuidae: Noctuinae).

The identity of Conisania leuconephra (Draudt, 1950) from Yunnan (China) is revised. A new species Conisania sejilaensis sp. nov. is described from Xizang (China). The adults as well as the male and the female genitalia of the new and related species are illustrated.

[Phylogenetic structure of Abies georgei var. smithii community at different spatial scales in Sygera Mountains of Tibet, China]. / 不同空间尺度西藏色季拉山急尖长苞冷杉群落谱系结构特征.

We analyzed the phylogenetic structure of trees within six diameter classes (1-2, 2-4, 4-7, 7-11, 11-16, >16 cm) in quadrats with different size of 5 m×5 m,10 m×10 m, 20 m×20 m, 50 m×50 m, 100 m×100 m in a Abies georgei var. smithii community in a 4 hm2 stem-mapping plot located in subalpine dark coniferous forest of Sygera Mountains, southeast Tibet. In various spatial scales, both net relatedness index (NRI) and nearest taxon index (NTI) of the community were larger than zero, indicating a clustered phylogenetic structure with the largest clustering intensity at small spatial scale (5 m×5 m). Community of small-size classes were phylogenetically clustering. In large-size classes (DBH>7 cm) phylogenetic over dispersion became more common, with dispersion increased with increasing tree size under all spatial scales. The intensity of phylogenetic clustering in young trees increased with increasing spatial scales, while the intensity of over dispersion in large trees (DBH>7 cm) increased with spatial scale. Our results suggested that environmental filtering in small-size trees and competitive exclusion in large-size trees might be the main ecological processes driving community assembly in this region.

The complete chloroplast genome of Abies georgei Orr var. smithii, a species endemic to the Qinghai-Tibet Plateau, China.

Abies georgei Orr var. smithii is an evergreen coniferous species of Pinaceae, and is endemic to the Qinghai-Tibet Plateau of China. Considering its vital ecological functions in this unique area, the complete chloroplast (cp) genome was constructed in this study to provide genetic information for its further study of conservation and evolution. The complete cp genome is 121,213 bp in length with GC content of 38.3%, and contains a tetrad structure, including a large single copy region of 76,278 bp, a small single copy of 42,575 bp, and two very short repeats of 1,180 bp for each. Besides, it contains 113 genes in total, including 74 CDSs, 35 tRNAs, and four rRNAs. This genome has been deposited in Genbank under accession number of MT527722.

[New records of medicinal plants in Tibet].

By the fourth survey of Chinese medicinal resources, new medicinal plants records of 2 genera and 5 species were reported in Tibet. They are two genera Rhynchoglossum and Asteropyrum, and five species including Rh. obliquum, A. peltatum, Urena repanda, Schefflera khasiana and Mimulus tenellus. All the voucher specimens are preserved in Herbarium of Tibet Agriculture and Animal Husbandry University.

Genetic diversity and population structure of two endemic Cupressus (Cupressaceae) species on the Qinghai-Tibetan plateau.

Cupressus gigantea and C. torulosa are ecologically and economically important endemic species of the conifer family Cupressaceae on the Qinghai-Tibetan plateau. C. gigantea was previously classified as a subspecies of C. torulosa because of their similar morphological characteristics and close distribution. In this study, 401 individuals were sampled from 16 populations of the two Cupressus species. The specimens were genotyped using 10 polymorphic microsatellite loci through fluorescence polymerase chain reaction (PCR). The genetic diversity of C. gigantea and C. torulosa populations was generally low, with the highest genetic diversity detected in the population LLS of C. gigantea. Distance-based phylogenetic and principal co-ordinates analyses indicated a clear genetic structures for the 16 populations of the two Cupressus species. Moreover, Mantel test results showed indistinctive correlations between population-pairwise Fst values and geographic distances, as well as between genetic distances and geographic distances in C. gigantea and C. torulosa, respectively. AMOVA suggested that genetic variation mostly resided within populations. Sixteen naturalpopulations were evidently clustered into two major groups in the constructed neighbour-joining tree. The results demonstrated that C. gigantea and C. torulosa are different Cupressus species. The genetic information provided important theoretical references for conservation and management of the two endangered Cupressus species.

Integrated Analysis of Transcriptomic and Proteomics Data Reveals the Induction Effects of Rotenoid Biosynthesis of Mirabilis himalaica Caused by UV-B Radiation.

Mirabilis himalaica (Edgew.) Heimerl is one of the most important genuine medicinal plants in Tibet, in which the special plateau habitat has been associated with its excellent medicinal quality and efficacy. However, the mechanisms by which environmental factors affect biosynthesis of secondary metabolic components remain unclear in this species. In this study, RNA sequencing and iTRAQ (isobaric Tags for Relative and Absolute Quantification) techniques were used to investigate the critical molecular "events" of rotenoid biosynthesis responding to UV-B radiation, a typical plateau ecological factor presented in native environment-grown M. himalaica plants. A total of 3641 differentially expressed genes (DEGs) and 106 differentially expressed proteins (DEPs) were identified in M. himalaica between UV-B treatment and control check (CK). Comprehensive analysis of protein and transcript data sets resulted in 14 and 7 DEGs from the plant hormone signal transduction and phosphatidylinositol signaling system pathways, respectively, being significantly enriched. The result showed that the plant hormone signal transduction and phosphatidylinositol signaling system might be the key metabolic strategy of UV-B radiation to improve the biosynthesis of rotenoid in M. himalaica. At same time, most of the DEGs were associated with auxin and calcium signaling, inferring that they might drive the downstream transmission of these signal transduction pathways. Regarding those pathways, two chalcone synthase enzymes, which play key roles in the biosynthesis of rotenoid that were thought as the representative medicinal component of M. himalaica, were significantly upregulated in UV-B radiation. This study provides a theoretical basis for further exploration of the adaptation mechanism of M. himalaica to UV-B radiation, and references for cultivation standardization.

Integrated analysis of transcriptomic and metabolomic data reveals critical metabolic pathways involved in rotenoid biosynthesis in the medicinal plant Mirabilis himalaica.

Mirabilis himalaica (Edgew.) Heimerl is among the most important genuine medicinal plants in Tibet. However, the biosynthesis mechanisms of the active compounds in this species are unclear, severely limiting its application. To clarify the molecular biosynthesis mechanism of the key representative active compounds, specifically rotenoid, which is of special medicinal value for M. himalaica, RNA sequencing and TOF-MS technologies were used to construct transcriptomic and metabolomic libraries from the roots, stems, and leaves of M. himalaica plants collected from their natural habitat. As a result, each of the transcriptomic libraries from the different tissues was sequenced, generating more than 10 Gb of clean data ultimately assembled into 147,142 unigenes. In the three tissues, metabolomic analysis identified 522 candidate compounds, of which 170 metabolites involved in 114 metabolic pathways were mapped to the KEGG. Of these genes, 61 encoding enzymes were identified to function at key steps of the pathways related to rotenoid biosynthesis, where 14 intermediate metabolites were also located. An integrated analysis of metabolic and transcriptomic data revealed that most of the intermediate metabolites and enzymes related to rotenoid biosynthesis were synthesized in the roots, stems and leaves of M. himalaica, which suggested that the use of non-medicinal tissues to extract compounds was feasible. In addition, the CHS and CHI genes were found to play important roles in rotenoid biosynthesis, especially, since CHS might be an important rate-limiting enzyme. This study provides a hypothetical basis for the screening of new active metabolites and the metabolic engineering of rotenoid in M. himalaica.

The complete chloroplast genome of Cupressus gigantea, an endemic conifer species to Qinghai-Tibetan Plateau.

The complete chloroplast genome of the wild Cupressus gigantea (Cupressaceae) is determined in this study. The circular genome is 128 244 bp in length with 115 single copy genes and two duplicated genes (trnI-CAU and trnQ-UUG). This genome contains 82 protein-coding genes, four ribosomal RNA genes and 31 transfer RNA genes. In these genes, eight genes (atpF, rpoC1, ndhA, ndhB, petB, petD, rpl16 and rpl2) harbor a single intron and two genes (rps12 and ycf3) harbor two introns. This genome does not contain canonical IRs, and the overall GC content is 34.7%. A maximum parsimony phylogenetic analysis revealed that C. gigantea and C. sempervirens are more closely related.

Characterization of the complete chloroplast genome of Juniperus cedrus (Cupressaceae).

The complete chloroplast genome of Juniperus cedrus is a circular molecule of 127 126 bp in length with 115 single copy genes and two duplicated genes (trnI-CAU and trnQ-UUG). The genome contains 82 protein-coding genes, four ribosomal RNA genes and 33 transfer RNA genes. In these genes, eight genes (rpl16, rpl2, rpoC1, petD, petB, ndhB, ndhA and atpF) harbor a single intron and two genes (rps12 and ycf3) harbor two introns. Like other sequenced chloroplast genomes of conifers, this genome does not contain canonical inverted repeats (IRs), and the overall GC content of J. cedrus chloroplast DNA is 35%. The phylogenetic analysis revealed that J. cedrus is more closely related to J. scopulorum and J. bermudiana.

Molecular cloning and transgenic characterization of the genes encoding chalcone synthase and chalcone isomerase from the Tibetan herbal plant Mirabilis himalaica.

Mirabilis himalaica is an endangered medicinal plant species in the Tibetan Plateau. The two genes respectively encoding chalcone synthase (MhCHS) and chalcone isomerase (MhCHI) were isolated and characterized from M. himalaica. The sequence analysis revealed that the two genes were similar with their corresponding homologous genes in other plants. The tissue profiles showed that both MhCHS and MhCHI had higher expression levels in roots than in stems and leaves. Transgenic hairy root cultures respectively with overexpressing MhCHS and MhCHI were established. The genomic PCR detection confirmed the authority of transgenic hairy root lines, in which either MhCHS or MhCHI expression levels were much higher than that in non-transgenic hairy root line. Finally, the HPLC detection results demonstrated that the rotenoid contents in MhCHS/MhCHI-transformed hairy root lines were enhanced. This study provided two candidate genes that could be used to genetic engineering rotenoid biosynthesis in M. himalaica and an alternative method to produce rotenoid using transgenic hairy root cultures.

[Spatial point pattern analysis of Abies georgei var. smithii in forest of Sygera Mountains in southeast Tibet, China].

In this study, based on a 4 hm2 stem-mapping plot, we analyzed spatial distributions of Abies georgei var. smithii, the dominant species in forest of Sygera Mountains in southeast Tibet, China. Pair-correlation function was used to characterize univariate spatial point patterns of three size classes of the population and bivariate spatial patterns between those and different sizes of dead wood. A. georgei var. smithii population was characterized by reverse J-shaped DBH distribution, indicating an increasing population. Saplings of the population were spatially obviously aggregated at the small scales (0-7 m), and mid-sized trees and large-sized trees of the population were randomly or uniformly distributed. The aggregation intensities of A. georgei var. smithii decreased with the increasing diameter classes and spatial scales. Saplings and mid-sized trees were significantly and negatively associated with large-sized trees at the small scales (0-35 and 0-30 m), but the associations reversed at the large scales (45-100 and 80-100 m). In addition, with the increasing age difference between diameter classes of the population, the intensities of positive or negative correlations increased. Spatial associations between saplings and dead large-sized trees, and between mid-sized trees and dead large-sized trees were negative at the small scales (0-34 and 5-27 m), but positive at the large scales (49-100 and 73-100 m). This suggested that released niche space due to dead large-sized trees is not enough to weaken their negative impacts on saplings. We concluded that self-thinning effect and Janzen-Connell hypothesis may be the main mechanisms for the spatial pattern formation of A. georgei var. smithii population.

[Study on medicinal plant resources and diversity in Rhinopithecus bieti national natural reserve of Markam in Tibet].

This research was a part of the investigation of traditional Chinese medicine resources survey in Markam. The medicinal plants in natural reserve were studied for the first in this paper. There were 300 species in 202 genera of 54 families, among them there were 7 species of ferns in 5 genera of 5 families, 6 species of gymnosperms in 4 genera of 3 families, and 287 species of angiosperms in 194 genera of 61 families. There were 166 species Tibetan medicinal plants in 102 genera of 47 families. Quantitative analysis was carried out in 6 aspects of family and genus composition, medicinal parts, drug properties, flavour of a drug, Tibetan medicine, toxicity and new plants. The concrete suggestions of protection and exploitation were put forward, which provided scientific basis for the sustainable utilization of medicinal plants in this area.

Engineering salidroside biosynthetic pathway in hairy root cultures of Rhodiola crenulata based on metabolic characterization of tyrosine decarboxylase.

Tyrosine decarboxylase initializes salidroside biosynthesis. Metabolic characterization of tyrosine decarboxylase gene from Rhodiola crenulata (RcTYDC) revealed that it played an important role in salidroside biosynthesis. Recombinant 53 kDa RcTYDC converted tyrosine into tyramine. RcTYDC gene expression was induced coordinately with the expression of RcUDPGT (the last gene involved in salidroside biosynthesis) in SA/MeJA treatment; the expression of RcTYDC and RcUDPGT was dramatically upregulated by SA, respectively 49 folds and 36 folds compared with control. MeJA also significantly increased the expression of RcTYDC and RcUDPGT in hairy root cultures. The tissue profile of RcTYDC and RcUDPGT was highly similar: highest expression levels found in stems, higher expression levels in leaves than in flowers and roots. The gene expressing levels were consistent with the salidroside accumulation levels. This strongly suggested that RcTYDC played an important role in salidroside biosynthesis in R. crenulata. Finally, RcTYDC was used to engineering salidroside biosynthetic pathway in R. crenulata hairy roots via metabolic engineering strategy of overexpression. All the transgenic lines showed much higher expression levels of RcTYDC than non-transgenic one. The transgenic lines produced tyramine, tyrosol and salidroside at higher levels, which were respectively 3.21-6.84, 1.50-2.19 and 1.27-3.47 folds compared with the corresponding compound in non-transgenic lines. In conclusion, RcTYDC overexpression promoted tyramine biosynthesis that facilitated more metabolic flux flowing toward the downstream pathway and as a result, the intermediate tyrosol was accumulated more that led to the increased production of the end-product salidroside.

[Biomass and yield of Betula platyphylla population in Mila Mount of Tibet].

Mila Mount in Southeast Tibet is the west boundary of the horizontal distribution of Betula platyphylla. It is also the highest altitude area in China and the world, where Betula platyphylla is distributed. Based on the investigation of regional sample plots and the analysis of analytic trees, this paper compared the biomass and yield of Betula platyphylla population at different slope orientation and different altitude. The results showed that the growth of Betula platyphylla was greatly interfered by human activities. The yearly productivity of Betula platyphylla grown in Mila Mount was 363.1-2,072.94 kg x hm(-2), and the biomass was 7,625.00-33,167.00 kg x hm(-2), indicating that the biomass of Betula platyphylla was quite different for different population. The productivity and biomass of Betula platyphylla coppice after cutting were lower than those of seedling forest, and there existed youth broken-down in the coppice. The zonal changes of water resources had no significant effect on the water allocation and the biomass of Betula platyphylla.