Ultrasonic-assisted extraction and adsorption separation: Large-scale preparation of trans-ε-Viniferin, suffruficosol B and trans-Gnetin H for the first time.

In this study, a Standard Operating Procedure (SOP) for the large-scale extraction, enrichment, and separation of suffruticosol B (SB), trans-ε-Viniferin (TV), trans-gnetin H (TG) from oil tree peony seeds shell (PSS) was successfully constructed. The ultrasonic-assisted extraction (UAE), macroporous adsorption resin (MAR), and column chromatography (CC) were employed to extract, enrich and separate SB, TV and TG from PSS, and the conditions were optimized. The results implied that SB (1.6937 g), TV (0.5884 g) and TG (3.8786 g) with the purity of 99.67 %, 99.32 % and 98.54 %, respectively, were obtained after the extraction, enrichment and separation. The total yields of the SB, TV and TG were 0.61 mg/g, 0.02 mg/g and 6.64 mg/g with the total extraction rates at 70.55 %, 69.77 % and 78.36 %, respectively. This is the first report on the large-scale extraction, enrichment and separation of oligostilbenes. The SOP in this paper could produce high purity SB, TV and TG, and provide a new idea for PSS as a new oligostilbene resource. The study expands the new development and research field of PSS and provides theoretical support for the green utilization of oil tree peony.

Major Fatty Acid Compositions and Antioxidant Activity of Cultivated Paeonia ostii under Different Nitrogen Fertilizer Application.

Paeonia ostii is now being extensively planted for oil extraction in China, which is recognized as a single oil-use tree peony cultivar and commonly called 'Fengdan'. This study investigated the effects of nitrogen fertilizer on oil yield, fatty acid compositions and antioxidant activity of P. ostii. Oil yield (33.46 %), oleic acid (25.12 %), linoleic acid (29.21 %) and α-linolenic acid (43.12 %) reached the maximum at N450 treatment, with significant differences compared with other treatments (P<0.05). Furthermore, strong antioxidant activity with low DPPHIC50 value (19.43±1.91 µg mL-1 ) and large ABTS value (1216.53±30.21 µmol Trolox g-1 ) and FRAP value (473.57±9.11 µmol Trolox g-1 ) was also observed at N450. Palmitic acid (5.57 %) and stearic acid (2.02 %) reached a maximum at N375, but not significant with N450 (P<0.05). Nitrogen fertilizer could promote oil yield, fatty acid accumulation and antioxidant activity, and N450 (450 kg ha-1 ) is recommended as the optimum application for P. ostii.

Major Fatty Acid Profiles and Bioactivity of Seed Oils from Ten†Tree Peony Cultivars as a Potential Raw Material Source for the Cosmetics and Healthy Products.

As a new woody oleaginous crop, tree peony is now being widely developed and utilized, which attributed to the outstanding oil-use features regarding the fatty acid profiles and bioactivity. The major fatty acid profiles and bioactivity of seed oils in ten tree peony varieties collected from a common garden were investigated in the present study. The results showed that the oil yields, fatty acid profiles, bioactivity of seed oils existed significant variations among ten tree peony varieties (P<0.05). Considered the application value, 'Fengdan' (FD) and 'Ziban' (ZB) was the optimal resources as the high oil yields, rich unsaturated fatty acids especially high α-linolenic acid of 40.46 %, great antioxidant activity with low IC50 values, high ABTS and FRAP values, and strong antimicrobial activity with high DIZ and low MIC/MBC values. The study also confirmed seed oil of tree peony as the potential raw materials sources in functional food, pharmaceuticals and cosmetics for human health.

Effects of plant population density and root-induced cytokinin on the corn compensatory growth during post-drought rewatering.

The effect of plant population density (PPD) and root-induced leaf cytokinin on the compensatory growth of potted corn seedlings during post-drought rewatering was investigated. The study design comprised four treatments: (1) wetness with low PPD, (2) wetness with high PPD, (3) rewatering with low PPD, and (4) rewatering with high PPD. Results showed that drought stress restrained the growth of corns. By contrast, rewatering enhanced the net photosynthetic rate and growth of corns. During the 8 days of rewatering, compensatory growth during post-drought rewatering occurred in corns with high PPD; however, such compensatory growth did not occur in corns with low PPD. Zeatin riboside concentrations in leaves and xylem saps were significantly higher under rewatering treatment than those under wet treatment. High leaf cytokinin concentration accelerated corn growth. The coefficients of variation and Gini-coefficient of wet treatment were significantly higher than those of rewatering treatment under high PPD, demonstrating that intense intraspecific competition occurred in the wet treatment. Extreme intraspecific competition negatively affected net photosynthetic rate. In brief, the interactions between root-induced leaf cytokinin and weak intraspecific competition promoted the compensatory growth under high PPD.

A simple and efficient method to isolate LTR sequences of plant retrotransposon.

Retrotransposons (RTNs) have important roles in the formation of plant genome size, structure, and evolution. Ubiquitous distributions, abundant copy numbers, high heterogeneities, and insertional polymorphisms of RTNs have made them as excellent sources for molecular markers development. However, the wide application of RTNs-based molecular markers is restricted by the scarcity of the LTR (long terminal repeat) sequences information. A new, simple, and efficient method to isolate LTR sequences of RTNs was presented based on the degenerate RNase H nested primers and PPT (polypurine tract) primer of RTNs in tree peony. This method combined the characteristics and advantages of high-efficiency thermal asymmetric interlaced PCR (hiTAIL-PCR), annealing control primer (ACP) system, and suppression PCR method. Nineteen LTR sequences were isolated using this new method in tree peony and the applicability of the LTR sequences based markers was validated by further SSAP analysis. The results showed that the new method is simple, of low-cost, and highly efficient, which is just conducted by three rounds of PCR and does not need any restriction enzymes and adapters, much less the hybridizations. This new method is rapid, economical, and cost- and time-saving, which could be easily used to isolate LTR sequences of RTNs.

Reverse transcriptase domain sequences from tree peony (Paeonia suffruticosa) long terminal repeat retrotransposons: sequence characterization and phylogenetic analysis.

Tree peony is an important horticultural plant worldwide of great ornamental and medicinal value. Long terminal repeat retrotransposons (LTR-retrotransposons) are the major components of most plant genomes and can substantially impact the genome in many ways. It is therefore crucial to understand their sequence characteristics, genetic distribution and transcriptional activity; however, no information about them is available in tree peony. Ty1-copia-like reverse transcriptase sequences were amplified from tree peony genomic DNA by polymerase chain reaction (PCR) with degenerate oligonucleotide primers corresponding to highly conserved domains of the Ty1-copia-like retrotransposons in this study. PCR fragments of roughly 270 bp were isolated and cloned, and 33 sequences were obtained. According to alignment and phylogenetic analysis, all sequences were divided into six families. The observed difference in the degree of nucleotide sequence similarity is an indication for high level of sequence heterogeneity among these clones. Most of these sequences have a frame shift, a stop codon, or both. Dot-blot analysis revealed distribution of these sequences in all the studied tree peony species. However, different hybridization signals were detected among them, which is in agreement with previous systematics studies. Reverse transcriptase PCR (RT-PCR) indicated that Ty1-copia retrotransposons in tree peony were transcriptionally inactive. The results provide basic genetic and evolutionary information of tree peony genome, and will provide valuable information for the further utilization of retrotransposons in tree peony.

[Identification and analysis methods of plant LTR retrotransposon sequences].

LTR retrotransposons are an important class of eukaryotic transposable elements, which are ubiquitous and highly heterogeneous in plant and play a major role in genome evolution of eukaryote. They are now extensively employed in gene function and genetic diversity analyses. Identification of LTR retrotransposons is the precondition for its application. Therefore, it has important theoretical significance and practical application value in studying identification and analysis methods LTR retrotransposon sequences. Bioinformatic software of the sequence analysis, according to the work principle, can be classified roughly into two types: sequence alignment and sequence identification of conserved domains. Alignment software, such as BLAST and DNAstar, produce the corresponding sequence information through comparison of sequence similarity; however, this kind of software cannot be applied for full length sequences. According to the principle, LTR retro-transposon sequence identification software can be roughly sorted into four types: de novo repeat discovery method, com-parative genomic method, homology-based method, and structure-based method. For example, LTR_Finder based on de novo repeat discovery method can accurately predict and annotate LTR retrotransposons for full length sequences; Repeat-Masker, which is based on homology-based method, can discover LTR retrotransposons by comparing the similarity with known sequences in the database. In this article, different methods of identification and analysis of retrotransposon se-quences were compared and analyzed, and a set of flow of LTR retrotransposons sequence analysis was summarized in order to provide the reference for LTR retrotransposons sequence analysis.

Isolation and characterization of 10 SSR markers of Momordica charantia (Cucurbitaceae).

PREMISE OF THE STUDY: Microsatellite markers were isolated and characterized from the genome of Momordica charantia (bitter melon) to be applied in studies of genetic diversity and population structure. METHODS AND RESULTS: Twenty-five microsatellite loci were isolated from the genome of bitter melon using the Fast Isolation by AFLP of Sequences COntaining Repeats (FIASCO) method. Ten loci were polymorphic, and the number of alleles per locus ranged from three to seven, with the observed heterozygosity ranging from 0.46 to 0.65. The markers also amplified successfully in the related species M. cochinchinensis and Cucurbita pepo. CONCLUSIONS: These markers will have potential utility for applications in genetic diversity evaluation, molecular fingerprinting, identification, comparative genomics analysis, and genetic mapping in Momordica species, as well as in C. pepo.

Development and characterization of EST-SSR markers in Paeonia suffruticosa (Paeoniaceae).

PREMISE OF THE STUDY: Paeonia suffruticosa is one of the most famous flowering species in China. Ten simple sequence repeat (SSR) markers were developed and characterized from the expressed sequence tag (EST) database of tree peonies to be applied in genetic diversity studies. METHODS AND RESULTS: A total of 10 novel EST-SSR loci were developed and characterized when tested on two populations of 45 tree peony individuals from China. The number of alleles per locus ranged from one to four and the expected heterozygosity ranged from 0.2087 to 0.6581. Select loci also amplified successfully in three other Paeonia species (P. ostii, P. rockii, P. jishanensis). CONCLUSIONS: These markers will facilitate analysis of genetic diversity, population differentiation, and marker-assisted breeding of Paeoniaceae.