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, Volume 9 Issue 5 Previous Issue    Next Issue
Plant Sciences
Transient expression of chicken alpha interferon gene in lettuce
Li SONG, De-gang ZHAO, Yong-jun WU, Yi LI
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2008, 9(5): 351-355.   https://doi.org/10.1631/jzus.B0710596
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We investigated the possibility of producing chicken alpha interferon (ChIFN-α) in transgenic plants. The cDNA encoding ChIFN-α was introduced into lettuce (Lactuca sativa L.) plants by using an agro-infiltration transient expression system. The ChIFN-α gene was correctly transcribed and translated in the lettuce plants according to RT-PCR and ELISA assays. Recombinant protein exhibited antiviral activity in vitro by inhibition of vesicular stomatitis virus (VSV) replication on chicken embryonic fibroblast (CEF). The results demonstrate that biologically active avian cytokine with potential pharmaceutical applications could be expressed in transgenic lettuce plants and that it is possible to generate interferon protein in forage plants for preventing infectious diseases of poultry.
Assessment of different genetic distances in constructing cotton core subset by genotypic values
Jian-cheng WANG, Jin HU, Xin-xian HUANG, Sheng-chun XU
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2008, 9(5): 356-362.   https://doi.org/10.1631/jzus.B0710615
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One hundred and sixty-eight genotypes of cotton from the same growing region were used as a germplasm group to study the validity of different genetic distances in constructing cotton core subset. Mixed linear model approach was employed to unbiasedly predict genotypic values of 20 traits for eliminating the environmental effect. Six commonly used genetic distances (Euclidean, standardized Euclidean, Mahalanobis, city block, cosine and correlation distances) combining four commonly used hierarchical cluster methods (single distance, complete distance, unweighted pair-group average and Ward’s methods) were used in the least distance stepwise sampling (LDSS) method for constructing different core subsets. The analyses of variance (ANOVA) of different evaluating parameters showed that the validities of cosine and correlation distances were inferior to those of Euclidean, standardized Euclidean, Mahalanobis and city block distances. Standardized Euclidean distance was slightly more effective than Euclidean, Mahalanobis and city block distances. The principal analysis validated standardized Euclidean distance in the course of constructing practical core subsets. The covariance matrix of accessions might be ill-conditioned when Mahalanobis distance was used to calculate genetic distance at low sampling percentages, which led to bias in small-sized core subset construction. The standardized Euclidean distance is recommended in core subset construction with LDSS method.
Characterization of high-yield performance as affected by genotype and environment in rice
Song CHEN, Fang-rong ZENG, Zong-zhi PAO, Guo-ping ZHANG
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2008, 9(5): 363-370.   https://doi.org/10.1631/jzus.B0710603
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We characterized yield-relevant characters and their variations over genotypes and environments (locations and years) by examining two rice varieties (9746 and Jinfeng) with high yield potential. 9746 and Jinfeng were planted in two locations of Shanghai, China, during 2005 and 2006. The results show that there was a large variation in grain yield between locations and years. The realization of high yield potential for the two types of rice was closely related to the improved sink size, such as more panicles per square meter or grains per panicle. Stem and leaf biomasses were mainly accumulated from tillering stage to heading stage, and showed slow decline during grain filling. Meanwhile, some photosynthetic characters including net photosynthesis rate (Pn), leaf area index (LAI), specific leaf area (SLA), fluorescence parameter (maximum quantum yield of PSII, Fv/Fm), chlorophyll content (expressed as SPAD value), as well as nutrient (N, P, K) uptake were also measured to determine their variations over genotypes and environments and their relationships with grain yield. Although there were significant differences between years or locations for most measurements, SLA at tillering and heading stages, Fv/Fm and LAI at heading stage, stem biomass at heading and maturity stages, and leaf nitrogen concentration at tillering and heading stages remained little changed, indicating their possible applications as selectable characters in breeding programs. It was also found that stem nitrogen accumulation at tillering stage is one of the most important and stable traits for high yield formation.
Interaction between seed size and NaCl on germination and early seedling growth of some Turkish cultivars of chickpea (Cicer arietinum L.)
Muharrem KAYA, Gamze KAYA, Mehmet Demir KAYA, Mehmet ATAK, Sevil SAGLAM, Khalid Mahmood KHAWAR, Cemalettin Yasar CIFTCI
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2008, 9(5): 371-377.   https://doi.org/10.1631/jzus.B0720268
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Chickpea is an important food legume crop of Turkey and is largely grown for human consumption on low moisture or salt-affected soils. The objective of the study was to find the effects of NaCl stress at electrical conductivities of 4.5, 8.6, 12.7 and 16.3 dS/m and seed sizes (7, 8 and 9 mm) on germination and early seedling growth of three popular chickpea cultivars (AKN-97, Gokce and Uzunlu-99). Mean frequency of germination, germination time, germination index, root length, shoot length and seedling fresh weight showed seed size-dependent responses of cultivars to salt stress. In general, small seeds germinated and grew more rapidly compared to medium and large seeds of the same cultivars against all levels of salt stress, with the best results in cultivar Uzunlu-99. No effect of NaCl treatments was observed on frequency of germination; however, a drastic decrease in early seedling growth was recorded at increased NaCl concentrations. Regression analysis results showed a significantly positive relationship (P<0.01) between seed size and mean germination time, whereas a significantly negative relationship was recorded between seed size and germination index, root length, shoot length. Moreover, linear regression values apparently confirmed that increased seed size in each cultivar affected decreased germination index, root and shoot lengths with enhanced mean germination time. Thus, it was concluded that the use of small seeds could considerably reduce the production costs of chickpea in salt-affected soils.
Quantifying biochemical variables of corn by hyperspectral reflectance at leaf scale
Qiu-xiang YI, Jing-feng HUANG, Fu-min WANG, Xiu-zhen WANG
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2008, 9(5): 378-384.   https://doi.org/10.1631/jzus.B0730019
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To further develop the methods to remotely sense the biochemical content of plant canopies, we report the results of an experiment to estimate the concentrations of three biochemical variables of corn, i.e., nitrogen (N), crude fat (EE) and crude fiber (CF) concentrations, by spectral reflectance and the first derivative reflectance at fresh leaf scale. The correlations between spectral reflectance and the first derivative transformation and three biochemical variables were analyzed, and a set of estimation models were established using curve-fitting analyses. Coefficient of determination (R2), root mean square error (RMSE) and relative error of prediction (REP) of estimation models were calculated for the model quality evaluations, and the possible optimum estimation models of three biochemical variables were proposed, with R2 being 0.891, 0.698 and 0.480 for the estimation models of N, EE and CF concentrations, respectively. The results also indicate that using the first derivative reflectance was better than using raw spectral reflectance for all three biochemical variables estimation, and that the first derivative reflectances at 759 nm, 1954 nm and 2370 nm were most suitable to develop the estimation models of N, EE and CF concentrations, respectively. In addition, the high correlation coefficients of the theoretical and the measured biochemical parameters were obtained, especially for nitrogen (r=0.948).
Biochemistry
Characterization of impurities in the bulk drug lisinopril by liquid chromatography/ion trap spectrometry
Pei-xi ZHU, Dan-hua WANG, Cui-rong SUN, Zhi-quan SHEN
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2008, 9(5): 385-390.   https://doi.org/10.1631/jzus.B0820031
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Two trace impurities in the bulk drug lisinopril were detected by means of high-performance liquid chromatography coupled with mass spectrometry (HPLC/MS) with a simple and sensitive method suitable for HPLC/MSn analysis. The fragmentation behavior of lisinopril and the impurities was investigated, and two unknown impurities were elucidated as 2-(6-amino-1-(1-carboxyethylamino)-1-oxohexan-2-ylamino)-4-phenylbutanoic acid and 6-amino-2-(1-carboxy-3-phenylpropylamino)-hexanoic acid on the basis of the multi-stage mass spectrometry and exact mass evidence. The proposed structures of the two unknown impurities were further confirmed by nuclear magnetic resonance (NMR) experiments after preparative isolation.
Synthetic and biological studies on a cyclopolypeptide of plant origin
Rajiv DAHIYA, Akhilesh KUMAR
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2008, 9(5): 391-400.   https://doi.org/10.1631/jzus.B0720001
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Objective: A natural cyclic peptide previously isolated from Citrus medica was synthesized by coupling of tetrapeptide units Boc-Leu-Pro-Trp-Leu-OMe and Boc-Ile-Ala-Ala-Gly-OMe after proper deprotection at carboxyl and amino terminals followed by cyclization of linear octapeptide segment. Methods: Solution phase technique was adopted for the synthesis of cyclooctapeptide—sarcodactylamide. Required tetrapeptide units were prepared by coupling of Boc-protected dipeptides viz. Boc-Leu-Pro-OH and Boc-Ile-Ala-OH with respective dipeptide methyl esters Trp-Leu-OMe and Ala-Gly-OMe. Cyclization of linear octapeptide unit was done by p-nitrophenyl ester method. The structure of synthesized cyclopolypeptide was elucidated by FTIR, 1H NMR, 13C NMR, FABMS spectral data and elemental analysis. The newly synthesized peptide was evaluated for different pharmacological activities including antimicrobial, anthelmintic and cytotoxic activities. Results: Synthesis of sarcodactylamide was accomplished with >78% yield utilizing dicyclohexylcarbodiimide (DCC) as coupling agent. Newly synthesized peptide possessed potent cytotoxic activity against Dalton’s lymphoma ascites (DLA) and Ehrlich’s ascites carcinoma (EAC) cell lines, in addition to moderate anthelmintic activity against earthworms Megascoplex konkanensis, Pontoscotex corethruses and Eudrilus sp. Moreover, cyclopolypeptide displayed good antimicrobial activity against pathogenic fungi Candida albicans and Gram-negative bacteria Pseudomonas aeruginosa, in comparison to standard drugs griseofulvin and ciprofloxacin. Conclusion: Solution phase technique employing DCC and triethylamine (TEA) as base proved to be effective for the synthesis of natural cyclooctapeptide. N-methyl morpholine (NMM) was found to be a better base for the cyclization of linear octapeptide unit in comparison to TEA and pyridine.
Kinetics of non-catalyzed hydrolysis of tannin in high temperature liquid water
Li-li LU, Xiu-yang LU, Nan MA
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2008, 9(5): 401-406.   https://doi.org/10.1631/jzus.B0730098
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High temperature liquid water (HTLW) has drawn increasing attention as an environmentally benign medium for organic chemical reactions, especially acid-/base-catalyzed reactions. Non-catalyzed hydrolyses of gallotannin and tara tannin in HTLW for the simultaneous preparation of gallic acid (GA) and pyrogallol (PY) are under investigation in our laboratory. In this study, the hydrolysis kinetics of gallotannin and tara tannin were determined. The reaction is indicated to be a typical consecutive first-order one in which GA has formed as a main intermediate and PY as the final product. Selective decomposition of tannin in HTLW was proved to be possible by adjusting reaction temperature and time. The present results provide an important basic data and reference for the green preparation of GA and PY.
Tannins from Canarium album with potent antioxidant activity
Liang-liang ZHANG, Yi-ming LIN
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2008, 9(5): 407-415.   https://doi.org/10.1631/jzus.B0820002
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The contents of total phenolics and extractable condensed tannins in the leaves, twigs and stem bark of Canarium album were determined. The structural heterogeneity of condensed tannins from stem bark was characterized by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and nuclear magnetic resonance (NMR) analyses. The results show the predominance of signals representative of procyanidins and prodelphinidins. In addition, epicatechin and epigallocatechin polymers with galloylated procyanidin or prodelphinidin were also observed. The tannins were screened for their potential antioxidant activities using 1,1-diphenyl-2-picrylhydrazyl (DPPH) and ferric reducing/antioxidant power (FRAP) model systems. Tannins extracted from leaves, twigs and stem bark all showed a very good DPPH radical scavenging activity and ferric reducing power.
Anaerobic ammonium oxidation for treatment of ammonium-rich wastewaters
Lei ZHANG, Ping ZHENG, Chong-jian TANG, Ren-cun JIN
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2008, 9(5): 416-426.   https://doi.org/10.1631/jzus.B0710590
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The concept of anaerobic ammonium oxidation (ANAMMOX) is presently of great interest. The functional bacteria belonging to the Planctomycete phylum and their metabolism are investigated by microbiologists. Meanwhile, the ANAMMOX is equally valuable in treatment of ammonium-rich wastewaters. Related processes including partial nitritation-ANAMMOX and completely autotrophic nitrogen removal over nitrite (CANON) have been developed, and lab-scale experiments proved that both processes were quite feasible in engineering with appropriate control. Successful full-scale practice in the Netherlands will accelerate application of the process in future. This review introduces the microbiology and more focuses on application of the ANAMMOX process.
Differential response of root morphology to potassium deficient stress among rice genotypes varying in potassium efficiency
Yan-bo JIA, Xiao-e YANG, Ying FENG, Ghulam JILANI
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2008, 9(5): 427-434.   https://doi.org/10.1631/jzus.B0710636
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Disparity in the root morphology of six rice (Oryza sativa L.) genotypes varying in potassium (K) efficiency was studied with three K levels: 5 mg/L (low), 10 mg/L (moderate) and 40 mg/L (adequate) in hydroponic culture. Morphological parameters included root length, surface area, volume and count of lateral roots, as well as fine (diameter<0.2 mm) and thick (diameter>0.2 mm) roots. The results indicate that the root growth of all genotypes was reduced under low K, but moderate K deficiency increased the root length of the efficient genotypes. At deficient and moderate K levels, all the efficient rice genotypes developed more fine roots (diameter<0.2 mm) than the inefficient ones. Both fine root count and root surface area were found to be the best parameters to portray K stress in rice. In accordance with the root morphology, higher K concentrations were noted in shoots of the efficient genotypes when grown at moderate and deficient K levels, indicating that root morphology parameters are involved in root uptake for K and in the translocation of K up to shoots. K deficiency affected not only the root morphology, but also the root ultra-structure. The roots of high-efficient genotypes had stronger tolerance to K deficient stress for root membrane damage, and could maintain the developed root architecture to adapt to the low K growth medium.
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