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Review
Induced pluripotent stem cells: origins, applications, and future perspectives
Jing Zhao, Wen-jie Jiang, Chen Sun, Cong-zhe Hou, Xiao-mei Yang, Jian-gang Gao
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2013, 14(12): 1059-1069.   https://doi.org/10.1631/jzus.B1300215
Abstract( 2061 )     PDF(0KB)( 1162 )
Embryonic stem (ES) cells are widely used for different purposes, including gene targeting, cell therapy, tissue repair, organ regeneration, and so on. However, studies and applications of ES cells are hindered by ethical issues regarding cell sources. To circumvent ethical disputes, great efforts have been taken to generate ES cell-like cells, which are not derived from the inner cell mass of blastocyst-stage embryos. In 2006, Yamanaka et al. first reprogrammed mouse embryonic fibroblasts into ES cell-like cells called induced pluripotent stem (iPS) cells. About one year later, Yamanaka et al. and Thomson et al. independently reprogrammed human somatic cells into iPS cells. Since the first generation of iPS cells, they have now been derived from quite a few different kinds of cell types. In particular, the use of peripheral blood facilitates research on iPS cells because of safety, easy availability, and plenty of cell sources. Now iPS cells have been used for cell therapy, disease modeling, and drug discovery. In this review, we describe the generations, applications, potential issues, and future perspectives of iPS cells.
Articles
Influence of heat stress on leaf ultrastructure, photosynthetic performance, and ascorbate peroxidase gene expression of two pear cultivars (Pyrus pyrifolia)
Dong-feng Liu, Dong Zhang, Guo-qin Liu, Sayed Hussain, Yuan-wen Teng
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2013, 14(12): 1070-1083.   https://doi.org/10.1631/jzus.B1300094
Abstract( 8489 )     PDF(0KB)( 1457 )
Plants encounter a variety of stresses in natural environments. One-year-old pot-grown trees of pear (Pyrus pyrifolia Nakai cv. Cuiguan and Wonhwang) were exposed to two heat stress regimes. Under constant short-term heat stress, chloroplasts and mitochondria were visibly damaged. Relative chlorophyll content and maximum photochemical efficiency of photosystem II were significantly decreased, which indicated that the leaf photosynthetic capability declined. Under chronic heat stress, mesophyll cell ultrastructure was not obviously damaged, but leaf photosynthetic capability was still restrained. As chronic heat stress was a simulation of the natural environment in summer, further study of the responses under this stress regime was undertaken. Ascorbate peroxidase (APX) activity was increased in ‘Cuiguan’, but not in ‘Wonhwang’. Inducible expression of PpAPX genes in the cytoplasm, chloroplasts and peroxisomes was consistent with increased APX activity in ‘Cuiguan’, whereas only weak induction of PpAPX genes was observed in ‘Wonhwang’. The isoenzymes cytosolic APX1 (cAPX1) and stromal APX (sAPX) were confirmed to be localized in the cytoplasm and chloroplasts, respectively.
Fungal diversity in adult date palm (Phoenix dactylifera L.) revealed by culture-dependent and culture-independent approaches
Ines Ben Chobba, Amine Elleuch, Imen Ayadi, Lamia Khannous, Ahmed Namsi, Frederique Cerqueira, Noureddine Drira, Néji Gharsallah, Tatiana Vallaeys
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2013, 14(12): 1084-1099.   https://doi.org/10.1631/jzus.B1200300
Abstract( 2616 )     PDF(0KB)( 2254 )
Endophytic flora plays a vital role in the colonization and survival of host plants, especially in harsh environments, such as arid regions. This flora may, however, contain pathogenic species responsible for various troublesome host diseases. The present study is aimed at investigating the diversity of both cultivable and non-cultivable endophytic fungal floras in the internal tissues (roots and leaves) of Tunisian date palm trees (Phoenix dactylifera). Accordingly, 13 isolates from both root and leaf samples, exhibiting distinct colony morphology, were selected from potato dextrose agar (PDA) medium and identified by a sequence match search wherein their 18S–28S internal transcribed spacer (ITS) sequences were compared to those available in public databases. These findings revealed that the cultivable root and leaf isolates fell into two groups, namely Nectriaceae and Pleosporaceae. Additionally, total DNA from palm roots and leaves was further extracted and ITS fragments were amplified. Restriction fragment length polymorphism (RFLP) analysis of the ITS from 200 fungal clones (leaves: 100; roots: 100) using HaeIII restriction enzyme revealed 13 distinct patterns that were further sequenced and led to the identification of Alternaria, Cladosporium, Davidiella (Cladosporium teleomorph), Pythium, Curvularia, and uncharacterized fungal endophytes. Both approaches confirmed that while the roots were predominantly colonized by Fusaria (members of the Nectriaceae family), the leaves were essentially colonized by Alternaria (members of the Pleosporaceae family). Overall, the findings of the present study constitute, to the authors’ knowledge, the first extensive report on the diversity of endophytic fungal flora associated with date palm trees (P. dactylifera).
Genetic variation in alkaloid accumulation in leaves of Nicotiana
Bo Sun, Fen Zhang, Guo-jun Zhou, Guo-hai Chu, Fang-fang Huang, Qiao-mei Wang, Li-feng Jin, Fu-cheng Lin, Jun Yang
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2013, 14(12): 1100-1109.   https://doi.org/10.1631/jzus.B1300130
Abstract( 1917 )     PDF(0KB)( 1459 )
Alkaloids are plant secondary metabolites that are widely distributed in Nicotiana species and contribute greatly to the quality of tobacco leaves. Some alkaloids, such as nornicotine and myosmine, have adverse effects on human health. To reduce the content of harmful alkaloids in tobacco leaves through conventional breeding, a genetic study of the alkaloid variation among different genotypes is required. In this study, alkaloid profiles in leaves of five Nicotiana tabacum cultivars and Nicotiana tomentosiformis were investigated. Six alkaloids were identified from all six genotypes via gas chromatograph-mass spectrometry (GC-MS). Significant differences in alkaloid content were observed both among different leaf positions and among cultivars. The contents of nornicotine and myosmine were positively and significantly correlated (R2=0.881), and were also separated from those of other alkaloids by clustering. Thus, the genotype plays a major role in alkaloid accumulation, indicating a high potential for manipulation of alkaloid content through traditional breeding.
Cloning and functional analysis of a novel ascorbate peroxidase (APX) gene from Anthurium andraeanum
Hui-chun Liu, Dan-qing Tian, Jian-xin Liu, Guang-ying Ma, Qing-cheng Zou, Zhu-jun Zhu
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2013, 14(12): 1110-1120.   https://doi.org/10.1631/jzus.B1300105
Abstract( 1747 )     PDF(0KB)( 1529 )
An 888-bp full-length ascorbate peroxidase (APX) complementary DNA (cDNA) gene was cloned from Anthurium andraeanum, and designated as AnAPX. It contains a 110-bp 5′-noncoding region, a 28-bp 3′-noncoding region, and a 750-bp open reading frame (ORF). This protein is hydrophilic with an aliphatic index of 81.64 and its structure consisting of α-helixes, β-turns, and random coils. The AnAPX protein showed 93%, 87%, 87%, 87%, and 86% similarities to the APX homologs from Zantedeschia aethiopica, Vitis pseudoreticulata, Gossypium hirsutum, Elaeis guineensis, and Zea mays, respectively. AnAPX gene transcript was measured non-significantly in roots, stems, leaves, spathes, and spadices by real-time polymerase chain reaction (RT-PCR) analysis. Interestingly, this gene expression was remarkably up-regulated in response to a cold stress under 6 °C, implying that AnAPX might play an important role in A. andraeanum tolerance to cold stress. To confirm this function we overexpressed AnAPX in tobacco plants by transformation with an AnAPX expression construct driven by CaMV 35S promoter. The transformed tobacco seedlings under 4 °C showed less electrolyte leakage (EL) and malondialdehyde (MDA) content than the control. The content of MDA was correlated with chilling tolerance in these transgenic plants. These results show that AnAPX can prevent the chilling challenged plant from cell membrane damage and ultimately enhance the plant cold tolerance.
Involvement of phosphatidate phosphatase in the biosynthesis of triacylglycerols in Chlamydomonas reinhardtii
Xiao-dong Deng, Jia-jia Cai, Xiao-wen Fei
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2013, 14(12): 1121-1131.   https://doi.org/10.1631/jzus.B1300180
Abstract( 1282 )     PDF(0KB)( 799 )
Lipid biosynthesis is essential for eukaryotic cells, but the mechanisms of the process in microalgae remain poorly understood. Phosphatidic acid phosphohydrolase or 3-sn-phosphatidate phosphohydrolase (PAP) catalyzes the dephosphorylation of phosphatidic acid to form diacylglycerols and inorganic orthophosphates. This reaction is integral in the synthesis of triacylglycerols. In this study, the mRNA level of the PAP isoform CrPAP2 in a species of Chlamydomonas was found to increase in nitrogen-free conditions. Silencing of the CrPAP2 gene using RNA interference resulted in the decline of lipid content by 2.4%鈥?7.4%. By contrast, over-expression of the CrPAP2 gene resulted in an increase in lipid content by 7.5%鈥?1.8%. These observations indicate that regulation of the CrPAP2 gene can control the lipid content of the algal cells. In vitro CrPAP2 enzyme activity assay indicated that the cloned CrPAP2 gene exhibited biological activities.
Morphological and biochemical responses of Oryza sativa L. (cultivar MR219) to ion beam irradiation
Anna Pick Kiong Ling, Ying Chian Ung, Sobri Hussein, Abdul Rahim Harun, Atsushi Tanaka, Hase Yoshihiro
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2013, 14(12): 1132-1143.   https://doi.org/10.1631/jzus.B1200126
Abstract( 1735 )     PDF(0KB)( 1445 )
Objective: Heavy ion beam, which has emerged as a new mutagen in the mutation breeding of crops and ornamental plants, is expected to result in the induction of novel mutations. This study investigates the morphological and biochemical responses of Oryza sativa toward different doses of carbon ion beam irradiation. Methods: In this study, the dry seeds of O. sativa were irradiated at 0, 20, 40, 60, 80, 100, and 120 Gy, followed by in-vitro germination under controlled conditions. Morphological and biochemical studies were conducted to investigate the morphological and physiological responses of O. sativa towards ion beam irradiation. Results: The study demonstrated that low doses (10 Gy) of ion beam have a stimulating effect on the height, root length, and fresh weight of the plantlets but not on the number of leaves. Meanwhile, doses higher than 10 Gy caused reductions in all the morphological parameters studied as compared to the control samples. The highest total soluble protein content [(2.11±0.47) mg/g FW] was observed in plantlets irradiated at 20 Gy. All irradiated plantlets were found to have 0.85% to 58.32% higher specific activity of peroxidase as compared to the control samples. The present study also revealed that low doses of ion beam (10 and 20 Gy) had negligible effect on the total chlorophyll content of O. sativa plantlets while 40 Gy had a stimulating effect on the chlorophyll content. Plantlets irradiated between 40 to 120 Gy were shown to be 0.38% to 9.98% higher in total soluble nitrogen content which, however, was not significantly different from the control samples. Conclusions: Carbon ion beam irradiation administered at low to moderate doses of 10 to 40 Gy may induce O. sativa mutants with superior characteristics.
Accumulation of mercury in rice grain and cabbage grown on representative Chinese soils
Chun-fa Liu, Cheng-xian Wu, Muhammad T. Rafiq, Rukhsanda Aziz, Dan-di Hou, Zhe-li Ding, Zi-wen Lin, Lin-jun Lou, Yuan-yuan Feng, Ting-qiang Li, Xiao-e Yang
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2013, 14(12): 1144-1151.   https://doi.org/10.1631/jzus.B1300004
Abstract( 1685 )     PDF(0KB)( 895 )
A pot culture experiment was carried out to investigate the accumulation properties of mercury (Hg) in rice grain and cabbage grown in seven soil types (Udic Ferrisols, Mollisol, Periudic Argosols, Latosol, Ustic Cambosols, Calcaric Regosols, and Stagnic Anthrosols) spiked with different concentrations of Hg (CK, 0.25, 0.50, 1.00, 2.00, and 4.00 mg/kg). The results of this study showed that Hg accumulation of plants was significantly affected by soil types. Hg concentration in both rice grain and cabbage increased with soil Hg concentrations, but this increase differed among the seven soils. The stepwise multiple regression analysis showed that pH, Mn(II), particle size distribution, and cation exchange capacity have a close relationship with Hg accumulation in plants, which suggested that physicochemical characteristics of soils can affect the Hg accumulation in rice grain and cabbage. Critical Hg concentrations in seven soils were identified for rice grain and cabbage based on the maximum safe level for daily intake of Hg, dietary habits of the population, and Hg accumulation in plants grown in different soil types. Soil Hg limits for rice grain in Udic Ferrisols, Mollisol, Periudic Argosols, Latosol, Ustic Cambosols, Calcaric Regosols, and Stagnic Anthrosols were 1.10, 2.00, 2.60, 2.78, 1.53, 0.63, and 2.17 mg/kg, respectively, and critical soil Hg levels for cabbage are 0.27, 1.35, 1.80, 1.70, 0.69, 1.68, and 2.60 mg/kg, respectively.
Sorption of ammonium and phosphate from aqueous solution by biochar derived from phytoremediation plants
Zheng Zeng, Song-da Zhang, Ting-qiang Li, Feng-liang Zhao, Zhen-li He, He-ping Zhao, Xiao-e Yang, Hai-long Wang, Jing Zhao, Muhammad Tariq Rafiq
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2013, 14(12): 1152-1161.   https://doi.org/10.1631/jzus.B1300102
Abstract( 2043 )     PDF(0KB)( 1744 )
The study on biochar derived from plant biomass for environmental applications is attracting more and more attention. Twelve sets of biochar were obtained by treating four phytoremediation plants, Salix rosthornii Seemen, Thalia dealbata, Vetiveria zizanioides, and Phragmites sp., sequentially through pyrolysis at 500 °C in a N2 environment, and under different temperatures (500, 600, and 700 °C) in a CO2 environment. The cation exchange capacity and specific surface area of biochar varied with both plant species and pyrolysis temperature. The magnesium (Mg) content of biochar derived from T. dealbata (TC) was obviously higher than that of the other plant biochars. This biochar also had the highest sorption capacity for phosphate and ammonium. In terms of biomass yields, adsorption capacity, and energy cost, T. dealbata biochar produced at 600 °C (TC600) is the most promising sorbent for removing contaminants (N and P) from aqueous solution. Therefore, T. dealbata appears to be the best candidate for phytoremediation application as its biomass can make a good biochar for environmental cleaning.
Atrazine biodegradation efficiency, metabolite detection, and trzD gene expression by enrichment bacterial cultures from agricultural soil
Robinson David Jebakumar Solomon, Amit Kumar, Velayudhan Satheeja Santhi
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2013, 14(12): 1162-1172.   https://doi.org/10.1631/jzus.B1300001
Abstract( 2071 )     PDF(0KB)( 1491 )
Atrazine is a selective herbicide used in agricultural fields to control the emergence of broadleaf and grassy weeds. The persistence of this herbicide is influenced by the metabolic action of habituated native microorganisms. This study provides information on the occurrence of atrazine mineralizing bacterial strains with faster metabolizing ability. The enrichment cultures were tested for the biodegradation of atrazine by high-performance liquid chromatography (HPLC) and mass spectrometry. Nine cultures JS01.Deg01 to JS09.Deg01 were identified as the degrader of atrazine in the enrichment culture. The three isolates JS04.Deg01, JS07.Deg01, and JS08.Deg01 were identified as efficient atrazine metabolizers. Isolates JS04.Deg01 and JS07.Deg01 produced hydroxyatrazine (HA) N-isopropylammelide and cyanuric acid by dealkylation reaction. The isolate JS08.Deg01 generated deethylatrazine (DEA), deisopropylatrazine (DIA), and cyanuric acid by N-dealkylation in the upper degradation pathway and later it incorporated cyanuric acid in their biomass by the lower degradation pathway. The optimum pH for degrading atrazine by JS08.Deg01 was 7.0 and 16S rDNA phylogenetic typing identified it as Enterobacter cloacae strain JS08.Deg01. The highest atrazine mineralization was observed in case of isolate JS08.Deg01, where an ample amount of trzD mRNA was quantified at 72 h of incubation with atrazine. Atrazine bioremediating isolate E. cloacae strain JS08.Deg01 could be the better environmental remediator of agricultural soils and the crop fields contaminated with atrazine could be the source of the efficient biodegrading microbial strains for the environmental cleanup process.
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