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Research Article
Characterization of Novel Chalcone Synthase Gene (CnCHS) from Chamaemelum nobile
Xiaomeng Liu, Xiangxiang Meng, Jiabao Ye, Weiwei Zhang, Jie Chang and Feng Xu
Background and Objective: Flavonoids are one kind of the main active ingredients in medicinal plant of Chamaemelum nobile (C. nobile). Chalcone synthase (CHS) is the first key enzyme and plays an important role in flavonoid biosynthesis pathway. The aim of the study was to clone the cDNA sequence of CnCHS from C. nobile for the first time using RT-PCR and carried out bioinformatics analysis. Materials and Methods: The seeds of C. nobile was obtained from the laboratory group's early preservation sown in a nutrition bowl (20×20 cm) after immersion and germination on November 10, 2016 and was grown at 25/18°C in a controlled growth chamber (16 h light/8 h dark). A pair of specific primers was designed according to C. nobile transcriptome data and the CnCHS gene was cloned from C. nobile by Reverse Transcription-Polymerase Chain Reaction (RT-PCR). The CnCHS gene as well as the protein sequence were analyzed using the online website of National Center for Biotechnology Information(NCBI), ProtParam and bio analysis software of Vector NTI 11.5, Clustal X2.0 and MEGA6. The expression of CnCHS gene in different tissues of C. nobile was studied by quantitative RT-PCR (qRT-PCR). Data were analyzed with one-way ANOVA using SPSS11.0 for Windows. Results: The cDNA of 1,482 bp CnCHS gene was cloned and named CnCHS (GenBank accession No. MF069253). The CnCHS contained a 1,197 bp open reading frame (ORF) encoding 399 amino acids. The predicted molecular weight and isoelectric point of the CnCHS protein were 43.64 kDa and 6.11, respectively. Multiple alignment analysis revealed that CnCHS protein sequence was highly homologous to CnCHS proteins of other plants. Phylogenetic analysis revealed that the CnCHS was most closely related to the CnCHS of Asteraceae, indicating that they share a common evolutionary ancestor. The CnCHS gene was expressed in all tested tissues of C. nobile and the expression level was the highest in flowers. Conclusion: The CnCHS gene was isolated and characterized, laying a foundation for further study of flavonoid biosynthesis pathway in C. nobile.
Research Article
Evaluate the Effect of Inhibiting Pathogenic Bacteria and Fungus of Eczematous Dermatitis and Antioxidant Activity of Phenolic from Qinglicao (Polygonum chinense L. Var. chinense)
Xudong Jiang, Weiguang Wang, Xiaoting Huang, Axiang Song, Lujiao Lu, Xiaoling Lu and Gang Wu
Background: QingLiCao has been used for the treatment of skin diseases such as eczematous dermatitis in China-Vietnam border area. However, the mechanism of treatment effect is still unclear. This study aimed to investigate the inhibitory effect of pathogenic bacteria and fungus of eczematous dermatitis and evaluate the antioxidant activity of phenolic contents from QingLiCao, will be helpful to further understand the mechanism of action. Materials and Methods: The in vitro phenolic antimicrobial activity was determined using disc diffusion and antioxidant activity was evaluated by spectrophotometry. Results: The experimental results showed that the phenolics from QingLiCao had excellent antimicrobial activity to pathogenic bacteria and fungus of eczematous dermatitis, similar results were observed for the antioxidant activity. Conclusion: The results support the proposition that phenolic from QingLiCao might be a promising anti-eczematous dermatitis natural active ingredients merit further investigations.
Research Article
Phylogenetic Analysis of Newcastle Disease Virus from Indonesian Isolates Based on DNA-Sequence of Fusion Protein-Encoding Gene
Martha Purnami Wulanjati, Nastiti Wijayanti and Aris Haryanto
Background and Objective: Newcastle disease (ND) is an infectious disease caused by ND virus (NDV), which is the major problem in poultry industries. Although vaccination program has been executed in Indonesia, Newcastle disease is still infecting chicken. The lack of vaccine protection against the disease, presumably due to genetic differences between vaccine strain and virus strain in the field. This study was conducted to analyze phylogenetic of Indonesian NDV isolates based on fusion (F) protein-encoding gene, with aim to determine which genotype group of Indonesian NDV isolates, compared to vaccine strain that circulating in Indonesia. Materials and Methods: Samples were RNA genome of NDV, which were isolated from chicken in Kartosuro, Karanganyar; Galur, Kulon Progo and Sukomoro, Magetan areas. The F gene was amplified by one step RT-PCR method and then was sequenced. The genetic variation among samples, other Indonesian isolates, LaSota and B1 vaccine strain were analyzed by pairwise distance method. Phylogenetic tree based on F gene sequence was generated by Neighbor-Joining method (1000 bootstrap repetition) and Kimura-2 parameter model. Result: The genetic variation of F gene nucleotide and F protein amino acid between samples and vaccine strains were higher compared to the variation between samples and other Indonesian isolates. Phylogenetic analysis showed that the samples were included in genotype VII class II, while LaSota and B1 vaccine strain included in genotype II class II. Conclusion: There were differences of fusion protein antigen and genotype group between Indonesian strains and vaccine strains. This finding encouraged to develop ND vaccine based on Indonesian isolates.
Research Article
De Novo Transcriptome Profiling of Buasbuas (Premna pubescens. Blume)
Diky Setya Diningrat and Erly Marwani
Background and Objective: Buasbuas is the one of the medicinal plants in Indonesia that contains bioactive compounds potential as antimicrobial, antioxidant, antidiabetes, antiinflammation and anticancer. Exploring the pathway and gene related of buasbuas bioactive compounds production has led to the renaissance of understanding buasbuas molecular mechanism database. The aim of this study was to developed data-mining framework of buasbuas to study plant specialized metabolism for phytochemical biosynthesis. Material and Methods: This project was started by collecting shoots and leaves of Buasbuas. Focus of the project was exploring the molecular mechanisms on biosynthesis phytochemical of Buasbuas. Illumina Mi-Seq Next Generation Sequencing was utilized to understand the molecular mechanisms of biosynthesis. Transcriptomes then trimmed and assembled with CLCBio genomic software. Assembled contigs then annotated towards Arabidopsis thaliana using CLC Bio genomic software. Digital Gene Expression was performed to analyze the transcriptional changes in control culture and treatment. Results: There were 5.342 unigenes that expressed only in treatment shoot cultures. Annotation with Gene Ontology showed that 57.9% (3.446) unigenes play role in Biological Process, 56.7% (3.375) unigenes play role in Cellular Components and 63.4% (3.772) unigenes play role in Molecular Functions. Annotation with Kyoto Encyclopedia of Genes and Genomes shows 853 unigenes essentially have role in 24 biological pathways. Highest process with highest unigenes involvement is biosynthesis of plant hormones and biosynthesis of alkaloids. Conclusion: This study showed that phytochemical biosynthesis in buasbuas induces level expression of several genes involved in the jasmonic acid, cytokinin, gibberellin, salicylic acid and ethylene biosynthesis pathway.
Research Article
Genetic Diversity and Population Structure of 29 Species of Genus Populus Assessed by AFLP Markers
Zhongcheng Zhou, Jiaping Yan, Xinghu Zhang, Zhongya Ye, Feng Xu and Jichi Yuan
Background and Objective: Populus species are one of the world’s most important groups of forest trees, but distinguishing between cultivars and hybrids is a little difficult due to widespread interspecific hybridization and introgression and the high level intraspecific morphological variation. The objective of this study was to understand the genetic variation and heterozygosity of 29 Populus species from the Jianghan Plain of China and provided scientific guidance for the introduction of superior poplar varieties in Hubei province of China. Materials and Methods: The DNA was isolated from leaves of 29 poplar clones, digested by MseI and EcoRI endonucleases and subsequently used Amplified Fragment Length Polymorphisms (AFLP) analysis. The AFLP data was analyzed by GeneMapper software to display the fragment sizes as electropherograms and binary data. Fingerprint was constructed according to the AFLP analysis. Genetic similarity coefficients were calculated using Numeric Taxonomic System (NTSYS) PC version software 2.11 and the phylogenetic tree was constructed by cluster analysis using the UPGMA. Results: A total of 959 fragments were amplified, 806 of which were polymorphic with an average polymorphism percentage of 84.67%. Genetic similarity coefficients of the 29 Populus species ranged from 0.6111-0.8478 with an average of 0.7217. The highest average genetic similarity coefficient (0.7549) was found between "Danhongyang" with other Populus species and the lowest (0.6532) was found between "6204" with other Populus species. According to the genetic similarity coefficient of 0.7117, the 29 Populus species were divided into four categories by UPGMA cluster analysis. Finally, we built DNA fingerprint of the 29 Populus genotypes. Conclusion: It was concluded that 29 Populus species shared high genetic similarity and low genetic diversity. "107", "6204" and "6102" were three unique species and worth further utilizing in hybridization.
Research Article
Oxalic Acid as the Main Molecule Produced by Trichoderma asperellum MG323528 Fermented on Corn Stover Based Medium
Abdulaziz Abdulrahman Al-Askar, WesamEldin Ismail Ali Saber, Khalid Mohamed Ghoneem and Younes Mohamed Rashad
Background and Objective: Organic acids have several pharmaceutical, food, agricultural and medical applications. Corn stover represents a serious environmental problem. The present study investigated the bio processing of such readily available low-cost biomass with microorganism into valuable organic acids that expected to neutralize the negative impact on the environment and minimize the production costs. Materials and Methods: A novel cellulolytic Trichoderma asperellum MG323528 was selected as a new corn stover decomposer that could transform it into various bio-products. The fungus was incorporated in corn stover-based medium for the production of organic acids. The Box-Behnken experimental design was applied to maximize the total organic acids production especially oxalic acid. Results: The optimum composition of solid-state fermentation medium was found to contain17.83 mg P from rock phosphate, 5.61 mg N from (NH4)2SO4 and 9.84 mg MgSO4•7H2O per 1 g of corn stover, yielding a total of 209.11±1.20 mmol organic acids. According to the HPLC screening, the main organic acids detected in the fermented corn stover was oxalic acid, representing about 78% of the total organic acids, in addition to minor amounts of citric, formic, salicylic and ascorbic acids. Conclusion: This kind of homo-fermentation could be considered for large-scale production of oxalic acid on an economic medium of CS using the promising T. asperellum MG323528 strain.

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