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[Identification of Salvia shandongensis new species based on sequences of the plastid psbA-trnH intergenic region]
Abstract
To identify Salvia shandongensis and its relatives at molecular level, the psbA-trnH intergenic region of three species including Salvia shandongensis, Salvia miltiorrhiza and S. miltiorrhiza f. alba were amplified and sequenced. Sequences were assembled with CodonCode Aligner. The K2P genetic distances between Salvia shandongensis and its relatives were calculated and UPGMA tree was performed by MEGA5.0. The results indicated that the lengths of psbA-trnH regions of Salvia shandongensis were about 391 bp, while the lengths of psbA-trnH regions of Salvia miltiorrhiza and S. miltiorrhiza f. alba were about 386 bp. The psbA-trnH sequences showed considerable variations between species and thus were revealed as a promising candidate for barcoding of Salvia shandongensis and its relatives. The intra-specific genetic distances of Salvia shandongensis were 0, while the intra-specific genetic distances of Salvia miltiorrhiza and S. miltiorrhiza f. alba were 0.002 and 0.001 respectively. Additionally, the genetic distance of Salvia shandongensis and Salvia miltiorrhiza ranged from 0.034 to 0.04, and the genetic distance of Salvia shandongensis and S. miltiorrhiza f. alba ranged from 0.005 to 0.008, the intra-specific genetic distances of Salvia shandongensis were much smaller than that of Salvia miltiorrhiza and S. miltiorrhiza f. alba; clustering results showed that there were obvious differences between Salvia shandongensis, Salvia miltiorrhiza and S. miltiorrhiza f. alba, which was consistent with morphological characteristics. This study not only firstly provides the scientific basis for establishing the taxonomy position in molecular level and revealing their genetic relationships of S. shandongensis, S. miltiorrhiza and S. miltiorrhiza f. alba; but also provides DNA molecular identification scientific basis for the development of new medicinal plant resources of Salvia shandongensis. Our results suggest that the psbA-trnH intergenic spacer region can be used as a barcoding to identify Salvia shandongensis, Salvia miltiorrhiza and S. miltiorrhiza f. alba.
Salvia miltiorrhiza Bunge is the source species of Danshen listed in Chinese Pharmacopoeia (2015 edition). Currently, the germplasm of S. miltiorrhiza and its related species are collected and preserved in several universities, research institutes, and companies where projects on Danshen have been conducted or are being conducted. Attempts have been made to use molecular markers and DNA barcoding in evaluating the genetic diversity and species identification of medicinal Danshen materials. Different cultivation modes, such as direct seeding and root propagation, are used in different production areas in China, and the latter is more popular than the former. From growing seedlings, transplanting, field management, root harvesting, to primary processing and storage, experiments have been conducted by researchers to establish efficient standard operating procedures (SOPs). Elite cultivars are crucial in elevating farming economic values. Bulk selection, system selection, cross-breeding, and space mutation breeding are adopted to obtain new cultivars. With the need for excellent cultivated varieties and the advancement of the application of next-/third-generation sequencing technologies, the preservation, evaluation, and utilization of S. miltiorrhiza and its related species will go even further, and new groundbreaking cultivars and breeding techniques will emerge. The nationwide standardization project of seed quality, plantation, reproduction, and crude medicinal material and decoction should promote the Danshen production industry. The release of the genome sequence of S. miltiorrhiza and related transcriptome analysis would accelerate germplasm evaluation and utilization and promote the breeding of elite cultivars.
A new species, Salvia lagochila T. Wang & L. Wang sp. nov. (Lamiaceae) is described from Shangri-La County in northwestern Yunnan, China. The diagnostic morphological characteristics in relation to closely similar species are discussed. Nutlet micro-morphology was examined using scanning electron microscopy, and leaf anatomic structures were studied using light microscope. The new species is included in S. subgen. Salvia sect. Eurysphace subsect. Perennes on the basis of its perennial habit, entire upper floral lip and arcuate anther connectives.
AIM: To study the correlation between ITS sequence variation of Cnidium monnieri and geographical distribution and chemo-type of the plant. METHODS: Two pairs of primers Pf + Pb and P5.8S ITS1+P5.8S ITS2 were designed. The PCR products were purified and sequenced by silver staining DNA sequencing technique and ABI 310. RESULTS: Complete sequence of ITS and 5.8S rDNA, and partial sequence of 18S rDNA and 26S rDNA of Cnidium monnieri were obtained. They totaled about 700 bp. The sequences of ITS-1 and ITS-2 ranged from 210 bp to 217 bp and 219 bp to 224 bp; respectively. Pairwese sequence divergences of the samples ranged from 0% to 1.93% in ITS-1, and from 0.46% to 2.34% in ITS-2. The ITS-1 is more conservative. Phylogenetic tree based on ITS-2 sequence data (225 aligned sites) indicated that the Harbin samples, Hengshui and Dezhou samples, and Zhengzhou and Gaochun samples clustered separately. CONCLUSION: The sequence variation of ITS-2 is correlated to latitude distribution of Cnidium monnieri in China. Further studies are needed to clarify the relation between ITS sequence variation and chemo-type of Cnidium monnieri.
