[Comparison of rDNA internal transcribed spacer sequences in asparagus].

OBJECTIVE: Using ITS sequence of nine species to identify counterfeiting medicine and analyse phylogenetic of Asparagus. METHODS: Analysing ITS sequences by amplification, cloning,sequencing and alignment. RESULTS: The length range of ITS sequence of nine species was from 711 to 748 bp, the percentage of G + C content was about 60%. The phylogenetic tree constructed on the basis of the ITS sequences showed that nine species were divided into two branches: Asparagus cochinchinensis, Asparagus officinalis, Asparagus densiflorus, Asparagus densiflorus cv. Myers and Asparagus densiflorus cv. Sprengeri were a branch and the others were a branch. Asparagus densiflorus and Asparagus densflorus cv. Myers those were from Africa had priority to clustering and then clustering with Asparagus densiflorus cv. Sprengeri that was a variant of Asparagus densiflorus in the first branch. Asparagus setaceus had relatively distant genetic relationship with the others three materials in another branch. CONCLUSIONS: The ITS sequences could distinguish species of Asparagus to test the counterfeit. Division status in phylogenetic tree of some species were debatable and ITS sequence was combined with others analytical tools to analyze the realistic phylogeny.

Pollen Typhae Total Flavone Inhibits Endoplasmic Reticulum Stress-Induced Apoptosis in Human Aortic-Vascular Smooth Muscle Cells through Down-Regulating PERK-eIF2α-ATF4-CHOP Pathway.

OBJECTIVE: To test the hypothesis that the inhibition of endoplasmic reticulum (ER) stress-induced apoptosis in oxidized low-density lipoproteins (ox-LDL)-induced human aortic-vascular smooth muscle cells (HA-VSMCs) was associated with suppression of the protein kinase RNA-like ER kinase (PERK)-eukaryotic translation initiation factor 2α (eIF2α)-activating transcription factor 4 (ATF4)-CCAAT/enhancer binding protein homologous protein (CHOP) signaling pathway by Pollen Typhae total flavone (PTF). METHODS: Primary HA-VSMCs were cultured and identified. The cultured HA-VSMCs were randomized into 5 groups, including a normal control group, an ox-LDL group (70 µg/mL high ox-LDL), an HPTF group (70 µg/mL high ox-LDL+500 µg/mL PTF), an MPTF group (70 µg/mL high ox-LDL+250 µg/mL PTF), and a LPTF group (70 µg/mL high ox-LDL+100 µg/mL PTF) in the first part; and a normal control group, an ox-LDL group (70 µg/mL high ox-LDL), an MPTF group (70 µg/mL high ox-LDL+250 µg/mL PTF), a shRNA group (transducted with PERK shRNA lentiviral particles), a scramble shRNA group (transducted with control shRNA lentiviral particles), an MPTF+ox-LDL+shRNA group (250 µg/mL PTF+70 µg/mL high ox-LDL+PERK shRNA lentiviral particles) and an ox-LDL+shRNA group (70 µg/mL high ox-LDL+PERK shRNA lentiviral particles) in the second part. The protein expression levels of ER-associated apoptosis proteins were detected by Western blot, and their mRNA expression levels were detected by quantitative real-time reverse transcription-polymerase chain reaction. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was applied to test cell viability, and the level of apoptosis was monitored by flow cytometry. RESULTS: The MTT assay and flow cytometry showed that the ox-LDL group had a significant increase in apoptosis, which was attenuated in PTF treatment groups and shRNA groups. Moreover, the ox-LDL group had increased protein and mRNA levels of binding immunoglobulin protein and ER-associated apoptosis proteins, such as PERK, eIF2α, ATF4 and CHOP, which were attenuated in PTF treatment groups and shRNA groups. CONCLUSIONS: The apoptosis induced by ox-LDL had a strong relation to ER stress. The protective effect of PTF on ER stressinduced apoptosis was associated with inhibition of the PERK-eIF2α-ATF4-CHOP pathway, which might be a potential therapeutic strategy for enhancing the stability of atherosclerotic plaques.

Comparison of chloroplast DNA between photoperiod-sensitive genic male sterile (PGMS) rice Nongken58S and its derivative sterile lines.

The ORF100, ORF29-TrnC(GCA) spacer, rps16 gene intron and TrnT(UGU)-TrnL(UAA) spacer of chloroplast DNA (cpDNA) of 6 photoperiod-sensitive genic male sterile (PGMS) rice, Nongken58S and its 5 derivatives, were amplified and sequenced. According to the result of ORF100 and ORF29-TrnC(GCA) spacer analysis, the cpDNA of japonica PGMS line Nongken58S was Japonica, and among those PGMS lines derived from Nongken58S, cpDNA of japonica 7001S and 3 indica lines 1103S, Peiai64S and Guangzhan63S were that of japonica, which was in accordance with the cytoplasm pedigree provided by their breeders. But the cpDNA of indica PGMS line W6154S was that of indica, which disaccorded with the cytoplasm pedigree, so we conjectured that the breeders had used the PGMS line as the male parent. Basing on the result of sequence analysis, we found single nucleotide polymorphism in rps16 gene intron and TrnT(UGU)-TrnL(UAA) spacer of these 5 japonica PGMS lines.

Photooxidation tolerance characters of a new purple pepper.

Huai Zi (HZ) is a new purple mutant of green pepper (PI 631133) that is obtained from the United States Department of Agriculture. The net photosynthetic rate (P N), chlorophyll fluorescence parameters, antioxidant substances, antioxidant enzymes, photosystem 1 (PS1) and PS2 activities were studied through methyl viologen (MV) treatment. The results showed that the P N, actual photochemical efficiency of PS2 (ΦPS2), photochemical quenching coefficient (qP), PS1 and PS2 activities in HZ were lower than those in green pepper. HZ had a stronger ability to eliminate reactive oxygen species(O2 (•-)) and accumulated less malondialdehyde (MDA) (a membrane lipid peroxidation product) than did green pepper, and had a higher content of antioxidants and antioxidant enzyme activity. This suggests that the lower light energy absorption and higher thermal dissipation and antioxidant activity of HZ contributed to a more stable PS2 photosynthetic capacity, which resulted in photooxidation tolerance. Hence, our study strongly suggests that pepper hybrids can achieve a modest ratio of chlorophyll and anthocyanin content, high P N and resistance to photooxidation, improving yield and resistance to adverse environments.