Table of Content

18 June 2019, Volume 36 Issue 3 Previous Issue    Next Issue

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Current applied researches of optical tweezers in biology LI Yin-mei, WANG Hao-wei, GONG Lei 2019, 36 (3):  1.  doi: 10.3969/j.issn.2095-1736.2019.03.001 Abstract ( 832 )   PDF   Save Invented 30 years ago, optical tweezers achieved a number of groundbreaking achievements in biology and promoted the development of biophysics. In 2018, A. Ashkin won the Nobel Prize in Physics for the invention of optical tweezers, which once again inspired the society′s attention to the technology of light. The ability of optical tweezers to quantitatively measure the force and actively manipulate micro-particles like cells makes it a unique tool in the biomedical field. This paper discusses optical tweezers from the perspective of biological applications. We hope it pave a way for biologists to address scientific questions and improve their sample preparation by revisiting the characteristics and functions of optical tweezers. Here we dedicated to popularize and advance existing research results by introducing examples of single beam optical tweezers applications that biologists can easily grasp. This paper focused on characteristics of physical biological cross-disciplinary research, and analyzed the technical bottlenecks involved in the study of single-cell and single-molecule by optical tweezers. We also introduced the research progress of optical tweezers technology and the current situation of the instrument market, looked forward to the future development and prospects of this field basing on our experience of optical tweezers. Related Articles | Metrics

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Cold atmospheric plasma-activated medium selectively inhibits triple negative breast cancer cells XIANG Liang-jian, XU Xiao-yu, ZHANG Shuo, CAI Dong-yan, DAI Xiao-feng 2019, 36 (3):  9.  doi: 10.3969/j.issn.2095-1736.2019.03.009 Abstract ( 418 )   PDF   Save Cold atmospheric plasma（CAP） has become a promising onco-therapeutic approach in recent years. Triple negative breast cancer is most aggressive among various breast cancer subtypes and has worst prognosis due to lack of effective therapy. We found that plasma-activated medium（PAM） selectively inhibited the proliferation and migration of triple negative breast cancer cells while causing little effect on other subtypes, and PAM selectively inhibited the phosphorylation of JNK in MDAMB231. We suggested that it was the more genome mutation and hyper-activated MAPK/JNK pathway in triple negative breast cancer cells that made them more sensitive to CAP. Our work revealed the relation between CAP and breast cancer subtypes and provides a preliminary basis for exploring the mechanism of CAP selectively killing triple negative breast cancer cells.  Related Articles | Metrics

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Construction and appraisal of recombinant adeno-associated virus vector for expression shRNA targeting NMU2R mRNA GUO Li-xia, YIN Zhong-yi, ZHENG Xu-xu 2019, 36 (3):  14.  doi: 10.3969/j.issn.2095-1736.2019.03.014 Abstract ( 330 )   PDF   Save To construct the recombinant adeno-associated virus vector carrying neuromedin U 2 receptor (NMU2R) short hairpin RNA (shRNA) for preparation of high-titer viruses, hairpin RNA was designed to target NMU2R rat mRNA, and was synthesized and cloned into pAAV-ZsGreen-shRNA plasmid which was double digested by BamH I and Hind III. The presence of the target sequence in the plasmid pAAV-ZsGreen-rNMU2R shRNA was confirmed by enzyme digestion, PCR and DNA sequencing analysis. And then the plasmid was transfected into AAV 293 cells. AAV 293 cells expression NMU2R shRNA were obtained and subsequently infected AAV packaging system to package the rAAV5-ZsGreen-rNMU2R shRNA. After purification, the functional and infectious virus was obtained and the titer of virus was 1×1012 vg/mL. Real-time PCR and western blot methods were used to detect the expression of NMU2R after infection with PC12 cells, and the empty viral vector pAAV-ZsGreen-shRNA was used as control. The results indicated that the expression of NMU2R mRNA and protein was decreased significantly after infection for 24 h and 48 h as compared with that of the control. In summary, a high-titer recombinant adeno-associated virus-5 vector carrying NMU2R shRNA has been constructed successfully. Related Articles | Metrics

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Construction and analysis of a yeast two-hybrid library responding to nitrogen in rice （Oryza sativa） HONG Guang-cheng, CHEN Qian, LIU Shi-feng, ZHANG Han-ma, QIN Xiao-jian 2019, 36 (3):  18.  doi: 10.3969/j.issn.2095-1736.2019.03.018 Abstract ( 304 )   PDF   Save Yeast two hybrid is an efficient molecular biology technique to study the interaction of proteins in eukaryotic yeast cells. In order to study the molecular regulation mechanism of nitrogen uptake and utilization in rice, we constructed a yeast two hybrid cDNA library in response to high nitrogen conditions by using nitrate (5 mmol/L KNO3) induced rice 9311 seedlings as materials. The test results showed that the titer of the library is 1.7×108 cfu/mL, the recombination rate of the library is 95% and the insert fragment size is 500-1500 bp. Furthermore, 10 clones were randomly selected from the library and followed sequencing, the results of NCBI alignment analysis showed that 9 clones could find the corresponding sequences from the rice gene pool. Summarily, the construction of this library helps to study the gene regulation of nitrogen absorption and utilization in rice. Related Articles | Metrics

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The divergence reason of black-crowned night heron(Nycticorax nycticorax) explored based on COX1 genes WANG Ye-ying, SU Hai-jun, XIONG Yong, ZHANG Ming-ming 2019, 36 (3):  22.  doi: 10.3969/j.issn.2095-1736.2019.03.022 Abstract ( 295 )   PDF   Save The process and reason of species divergence is one of the hot issues in evolutionary biology and ecology field, the nature of species divergence is the gene differentiation. This research studied the globally distributed black-crowned night heron. A phylogenetic tree was constructed by using the method of bayesian and maximum likelihood based on mitochondrial COX1 gene. A time tree was built by the BEAST software using a relaxed molecular clock and three calibration points from the divergence time 68.92 million years between Ciconiiformes and Pelecaniformes, 67.12 million years between Ciconiiformes and Sphenisciformes, 72.25 million years between Ciconiiformes and Sphenisciformes. The purpose of this study is to use molecular methods to evaluate the divergence time of black-crowned night heron and infer the divergence factors. The results showed that the night heron could be divided into two lineages according to their distribution areas, the Americas lineage and Asia lineage. The divergence of the two lineages approximately happened 56.74 million years ago. The separation of the Asian continent and America caused this divergence.  Related Articles | Metrics

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Genetic variation analysis of artificially introduced Bombina orientalis populations in Beijing ZHU Guo-fen, SHI Yang, TIAN Heng-jiu, LI Yang, GAO Fu-li, BAO Wei-dong 2019, 36 (3):  27.  doi: 10.3969/j.issn.2095-1736.2019.03.027 Abstract ( 371 )   PDF   Save The existing Oriental fire-bellied toads (Bombina orientalis) in Beijing are the descendant of more than 200 individuals introduced from Shandong populations in 1927. There were no new individuals introduced along the 89 years of independent evolution, therefor making this population become a good sample of studying wild animal population genetics. In this study, we reported the genetic differentiation of 55 Oriental fire-bellied toad individuals from Beijing and Shandong populations based on mitochondrial control region and 6 microsatellite loci. Mitochondrial markers showed the haplotype diversity of Beijing and Shandong populations were 0.489 and 0.950, and nucleotide diversity were 0.0018 and 0.0033, respectively. Phylogenetic trees and network of haplotypes indicated that the two populations have not genetically separated. Microsatellite markers showed the observed heterozygosity were 0.468 and 0.631, and expected heterozygosity were 0.548 and 0.701 for Beijing and Shandong populations, respectively. Structure analysis categorized the 54 successfully amplified individuals into Beijing and Shandong genetic clusters. In general, the Beijing population showed relatively low genetic diversity, which indicated a trend of genetic isolation draw back, but it was still at a moderate level for maintaining normal existence. Thus, there is no need to reintroduce new individuals from Shandong population. Related Articles | Metrics

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Influence of inter-planting with Rubus chingii on Castanea mollissima soil bacterial structure and diversity XU Yang, GONG Bang-chu, WU Kai-yun, JIANG Xi-bing, SUN Wei-min, HU Wei-bin, CHEN Xi-liang 2019, 36 (3):  31.  doi: 10.3969/j.issn.2095-1736.2019.03.031 Abstract ( 280 )   PDF   Save In order to provide a scientific basis for evaluating the management measure of Castanea mollissima inter-planting with Rubus chingii, the impact of soil bacterial structure and biodiversity was studied. Soil samples from Castanea mollissima inter-planting with different density of Rubus chingii (0, 100 and 400 plants/Mu）were collected. Soil bacteria were sequenced by Illumina High Throughput Sequencing technique. And, the soil physical and chemical indexes were also analyzed. The results showed that soil bacterial structure of Castanea mollissima forest was similar among different modes, Proteobacteria (31.20%) and Acidobacteria (29.70%) were the most common bacteria. After inter-planting with Rubus chingii, Chao1 and Simpson index indicated dominance of soil bacteria were increased significantly, but diversity was decreased. The influences of inter-planting and density were mainly affected by soil bacterial community on the Phylum level and Order level, respectively. Castanea mollissima forest soil organic matter, total nitrogen, hydrolyzable nitrogen, available phosphorus and total phosphorus content were increased significantly by inter-planting with Rubus chingii. The soil potassium content was decreased with the increase of inter-planting density. The changes of soil physical and chemical property caused by inter-planting may explain the influence of bacterial community structure. The changes of Acidobacteria and Acidobacteriales were correlated with nitrogen content. The change of Chloroflexi was correlated with organic matter.  Related Articles | Metrics

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Phytoplankton community structure of coal mining subsided area with environmental drivers in Linhuan Lake, Huaibei WANG Chen-chen, WU Qi-li, WAN Yang, LIN Zhi1, XU Jian, ZHOU Zhong-ze 2019, 36 (3):  37.  doi: 10.3969/j.issn.2095-1736.2019.03.037 Abstract ( 379 )   PDF   Save The phytoplankton community structure of Linhuan Lake in Huaibei City was conducted from August 2015 to June 2016, phytoplankton community structure and its relationships with environmental factors were analyzed. A total of 135 phytoplankton species belonging to 7 phyla, 65 genera were recorded. The phytoplankton community structure dominated by Cyanophyta, Chlorolhyta respectively, in which Cyanophyta and Chlorolhyta accounted for 64.93% of the total community composition. According to the abundance and occurrence, Cryptomonas erosa, Oscillatoria chlorine and Phormidium tenue were considered as the dominant species. The mean abundance and biomass of phytoplankton were 107.32×105 cells/L and 8.11 mg/L respectively, with the spatial pattern of decreasing from east to west. The Shannon-Wiener index(H′), Margalef index(D)， and Pielou index(J) of the phytoplankton community were 2.81, 2.96, and 0.617, respectively. The results of Pearson correlation analyses and the RDA indicated that water temperature, transparency, nutritive salt and chla were the four main environmental drivers affecting the phytoplankton community structure in Linhuan Lake. Combined with the existing research results, the succession characteristics of phytoplankton communities in lakes with different subsidence time series were discussed. Related Articles | Metrics

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Effect of heavy metal compound pollution on Cd uptake and accumulation in rice LIANG Rui,CHEN Hui-ru,LIU Bin-mei,YANG Yang,WU Yue-jin 2019, 36 (3):  42.  doi: 10.3969/j.issn.2095-1736.2019.03.042 Abstract ( 314 )   PDF   Save As heavy metal pollution in nature is dominated by multiple heavy metal pollution, the current study investigated the Cd uptake, transport and grain accumulation of a low-Cd-accumulation rice mutant (T2-1), based on field and hydroponics experiments. The results showed that Cd accumulation was significantly lower in T2-1 than in the wild type 9311. Compared with 9311, Cd accumulation in T2-1 with a single Cd-contaminated field was significantly reduced by 29.72%-61.63%. However, the content of Cd in grains of T2-1 planted on the plots contaminated by multiple heavy metals was reduced by only 11.07%. The results indicated that other heavy metals affected the absorption and transfer of Cd by T2-1 under the conditions of multiple heavy metal pollution. Under hydroponic conditions, seedling height and dry weight decreased with the addition of Pb and Cu. Compared with Cd and Cd-Pb treatments, addition of Cu had more significant effects on inhibiting rice growth. The trend of seedling height and dry weight were the same at 15 and 30 d. Dry matter accumulation of T2-1 was slightly smaller than 9311, which might be related to the energy consumption of fixing more Cd in roots of T2-1. The accumulation of Cd in root of T2-1 was greater than that of 9311, while that Cd transfer from root to shoot was lower in T2-1 than in 9311. Low concentration of Pb significantly promoted Cd uptake by rice seedlings, indicating that low concentration of Pb enhance Cd bioavailability in rice. In the Cd-Cu and Cd-Pb-Cu treatments, the contents of Cd in shoots and roots of two rice seedlings decreased significantly compared with those under Cd stress alone. Cu can inhibit the transfer of Cd in T2-1, and the inhibitory effect is much greater than the promotion effect of Pb. Overall, application of low-Cd-accumulation rice mutant has a promising option to alleviate Cd pollution in rice production and enhance food security. Whereas, as multiple heavy metal pollution affects the absorption, transportation and accumulation of Cd in rice，the identification of multiple heavy metal pollution should be considered in the screening of low-Cd-accumulation cultivars. Related Articles | Metrics

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Effects of the growth of Nannochloropsis XJ006 on the removal efficiency of four heavy metals ZHENG Kun-can,LI Yan-ran,WU Di,QIAN Gai-gai,LIU Jia,GONG Dong-hui,JI Xiang 2019, 36 (3):  47.  doi: 10.3969/j.issn.2095-1736.2019.03.047 Abstract ( 330 )   PDF   Save As a biosorbent adsorption of heavy metal elements， microalgae have the following advantages of environmental protection, efficient and easy to dissolve. In this paper, Nannochloropsis XJ006 was used as the heavy metal adsorbent. The effects of different concentrations of heavy metals Pb2+, Cd2+, Hg2+ and As3+ on Nannochloropsis XJ006 growth, and the removal rate of four heavy metals in different concentrations were studied. The results showed that when the Pd2+, Cd2+, Hg2+ and As3+ was at the concentration rang of 0-9 mg/L, 0-5 mg/L, 0-0.15mg/L and 0-2.5 mg/L, respectively, Nannochloropsis XJ006 can grow normally. The removal rate of Cd2+ was 88.40%, Pd2+, Hg2+ was 80% and As3+ was 40.04%. The orthogonal experiment showed that under the concentration of Pd2+ 5 mg/L, Cd2+ 1 mg/L, Hg2+ 0.05 mg/L and As3+ 0.5 mg/L, the algae grows best. Related Articles | Metrics

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The morphological and ultrastructural profiling between floating and benthic Sargassum horneri#br# ZHU Qi, REN Ji-rui, CHEN Juan-juan, YANG Rui 2019, 36 (3):  51.  doi: 10.3969/j.issn.2095-1736.2019.03.051 Abstract ( 307 )   PDF   Save The seaweed Sargassum horneri is an marine benthic brown alga in the Northwest Pacific and discontinuous distributed along the coast of China. The morphological traits of S. horneri are brown and consist of the holdfast, stem, leaves, gas vesicles and receptacles. There are two ecological types of S. horneri, the floating type and the benthic type. Comparing the morphological traits of each ecological type, significant differences were found. However, when it came to the microscopic and submicroscopic structures, almost no variation was found between these two types. Compared with benthic S. horneri, the floating S. horneri had shorter burl distance and more densities of burls and gas vesicles， which might be influenced by the external environment and internal physiology in the process of floating. There were three kinds of cells that were observed in the algae, which were epidermis, cortex and medulla cells from the surface to the center. The size of the cells were gradually increased from epidermis, cortex to medulla cells; while the contents of chloroplast (C)and physode (Ph)decreased stepwise, especially, there existed abundant of plasmodesmata (Pla)in the intercellular of cortical cells. Related Articles | Metrics

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Evaluation of cold resistance and physiological response to low temperature on five kinds of Ilex aquifolium CUI Ying-wen,LI Wen-ming, QIAN Yan-ping,TIAN Ru-nan, LI Yun-long 2019, 36 (3):  55.  doi: 10.3969/j.issn.2095-1736.2019.03.055 Abstract ( 497 )   PDF   Save In this paper，two years old seedlings of Ilex aquifolium，two hybrids of Ilex aquifolium (Ilex × altaclerensis‘Belgica Aurea’，Ilex × altaclerensis‘Lawsoniana’) and two varieties of Ilex aquifolium (Ilex aquifolium.‘Madame Briot’，Ilex aquifolium‘Little Gem’) were placed in different low temperature conditions， the morphological characteristics，relative conductivity，the activity of superoxide (SOD)，the level of malondialdehyde (MDA)，the contents of soluble sugar and chlorophyll were studied. Comprehensive evaluation of cold resistance on five kinds of plants was conducted by principal component analysis and subordinate function analysis. The results revealed that cold resistance of five kinds of plants from strong to weak was Ilex×altaclerensis ‘Belgica Aurea’>Ilex aquifolium ‘Little Gem’>Ilex aquifolium>Ilex aquifolium. ‘Madame Briot’>Ilex×altaclerensis ‘Lawsoniana’. Related Articles | Metrics

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Seasonal variations of sex hormones in Chinese goral at the Saihanwula Nature Reserve, Inner Mongolia#br# ZHANG Rui, CHEN Zhao-juan, JIANG Xiu-li, WANG Xiao-ling, WU Li-ji,MENGHE Da-lai, CHEN Chen, HAN Ying-ying, BAO Wei-dong 2019, 36 (3):  60.  doi: 10.3969/j.issn.2095-1736.2019.03.060 Abstract ( 307 )   PDF   Save The animal sex hormones are closely related to its reproduction activity. To obtain the seasonal variation characteristics of fecal sex hormones level of the Chinese goral (Naemorhedus griseus), we analyzed 554 fecal samples collected in 2015 and 2016 by using radioimmunoassay, while 20 female and 20 male fecal samples collected in 2013 were used as comparison from the Inner Mongolia Saihanwula Nature Reserve. The results showed that testosterone had the highest level in July(236.21 ng/g +23.31 ng/g, Mean+SD). And it also showed a high level in April and had no significantly difference between April 2016 and September 2013 (P>0.05). The progesterone showed the highest level in May (398.24 ng/g+39.11 ng/g) and was significantly higher than in other months (P<0.05). The estradiol levels showed the highest in March (36.36 ng/g+3.18 ng/g) and was also significantly higher than other months (P<0.05), but it showed the lowest level in July. The seasonal variations of sex hormones confirmed the document′s record that the rut season of the Chinese goral is in October and farrowing period is in May, and also confirmed the method of using radio immunoassays to detecte fecal hormones concentration levels of Chinese goral for analysis its seasonal changes is ideal. Related Articles | Metrics

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Study on the cause and control of Carapace ulcer of Litopenaeus vannamei in factory freshwater aquaculture#br# FAN Hui-min, MA Qian-qian, SHANG Ze-hao, ZHANG Zi-han, DONG Zhao-ran, TANG Qing-quan1, XU Xiao-mu, WANG Li-ming,ZHU Ruo-lin, BAO Chuan-he, PENG Kai-song, WAN Quan 2019, 36 (3):  65.  doi: 10.3969/j.issn.2095-1736.2019.03.065 Abstract ( 427 )   PDF   Save In order to determine the pathogen of the Carapace ulcer of Litopenaeus vannamei in factory freshwater aquaculture and provide the methods of prevention and treatment, analysis of isolation and identification of pathogens, virulence genes, toxicity test, the influence of phase related antibiotics and topical disinfectants on the aquatic animal were couducted. Samples were collected and isolated from the hepatopancreas of diseased shrimps and attacked water, and cultured with TCBS medium, screening for dominant strains of each group, respectively called F-L and F-W. Each group was identified based on the culture characteristics and forms, sequences of 16S rRNA gene and biochemical identification. The result of the F-L was Aeromonas veronii, and of the F-W was Aeromonas punctata. In virulence genes test, F-L strains detected 8 genes, respectively were aerA, Act, Lip, Fla, Exu, Ser, Alt and ahyB, and F-W strains had aerA, Alt, Lip, Fla, ahyB and hlyA. In toxicity test, both strains of bacteria could cause the death of Litopenaeus vannamei, and median lethal dose was 107.68 and 107.8 cfu/mL, respectively. The study would provide a basis of the effective prevention and control of Carapace ulcer of Litopenaeus vannamei caused by Aeromonas. Related Articles | Metrics

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The reasons for the difference of xanthohumol  content in different kinds of beer ZHAO Hui-xin, ZHOU Qian, ZENG Wei-jun, LU Han, ZHAO He-ping 2019, 36 (3):  70.  doi: 10.3969/j.issn.2095-1736.2019.03.070 Abstract ( 391 )   PDF   Save The aim of this study is to explore the main causes of different xanthohumol contents in different breeds. Six different beer and similar color dilution black ink shading experiments were conducted to compare their effects on the photoisomerization of xanthohumol samples. The results showed that any kind of beer covered or mixed into the sample could inhibit the photoisomerization of xanthohumol, and the inhibition effect of dark beer was better than that of light beer. In addition, the use of diluted black ink can also inhibit the photoisomerization of xanthohumol samples. The effect is consistent with the black beer with similar chromaticity. When adding different kinds of beer to xanthohumol samples and covering them with the same thickness of black ink, all varieties of beer could inhibit the decreasing of OD370, but there was no significant difference between all kinds of beer. And it implied that the real reason for the difference of xanthohumol content in different beer is that different brewers have different light transmittance, rather than the small molecule inhibitors in beer.  Related Articles | Metrics

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Research progress in regulation and function of cold induced RNA binding protein WANG Xiao-hui,XU Xiao-na,QIN Xu-jun,WANG Feng 2019, 36 (3):  74.  doi: 10.3969/j.issn.2095-1736.2019.03.074 Abstract ( 654 )   PDF   Save Cold induced RNA binding protein (Cirbp) is an evolutionary conservative RNA binding protein, including a RNA domain (RRM) and a arginine - glycine (RGG) domain. The expression of Cirbp changes in temperature, hypoxia, radiation, drugs, growth factors and so on, so as to regulate transcriptional, post transcriptional, and translation. This article reviews the evolutionary conservatism and intermolecular interaction, cell function and role in different physiological and pathological processes, including circadian rhythm, inflammation, nerve plasticity, stem cell characteristics and cancer. Related Articles | Metrics

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The research progress in Drosophila JNK signaling pathway TANG Cheng-chen， JIA Jia 2019, 36 (3):  78.  doi: 10.3969/j.issn.2095-1736.2019.03.078 Abstract ( 523 )   PDF   Save As a model organism, fruit flies play an extremely important role in embryonic development, pathogenesis of diseases and regulation of gene expression. When the fruit flies are stimulated by stressors, the cells in the body respond to a series of stress responses to maintain homeostasis. JNK signaling pathway plays a key role in fruit flies stress reaction. In this paper, combining with the latest research progress of related protein interactions from the JNK signaling pathway and negative regulatory factors of regulation, the stress response caused by the JNK signaling pathway related function were summarized. Related Articles | Metrics

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The research progress in the function and regulation of STAYGREEN in crops CHEN Yao-yu, WANG Shu-guang, SUN Dai-zhen 2019, 36 (3):  83.  doi: 10.3969/j.issn.2095-1736.2019.03.083 Abstract ( 698 )   PDF   Save STAYGREEN（SGR）/ NON-YELLOWING（NYE）gene encods a leaf senescence related protein, which plays an essential role in chlorophyll breakdown. Mutants of the SGR gene exhibit stay-green phenotype in many plants, however, the functions of different SGRs are variant. In this article, the latest research findings of SGR were reviewed from three aspects, including the function of SGR, the relationship between SGR and chlorophyll degradation, and the regulation of SGR. Meanwhile, the research direction of SGR in main grain crops in China was prospected. Related Articles | Metrics

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Roles of apoplastic protein in plant heavy metal tolerance RUI Hai-yun, LIU Qing-quan, SHEN Zhen-guo 2019, 36 (3):  88.  doi: 10.3969/j.issn.2095-1736.2019.03.088 Abstract ( 381 )   PDF   Save Apoplast is the plant cell compartment external to the plasma membrane which includes the cell walls, the intercellular space and the apoplastic fluid. Apoplast mainly consists of polysaccharides (cellulose, hemicelluloses and pectins), proteins and lignin, and has been demonstrated to be involved in response to different abiotic and biotic stresses. The paper summarizes the roles of apoplastic structural proteins, polysaccharide metabolism proteins, oxidation-reduction proteins and stress-responsive proteins in plant heavy metal tolerance to give a comprehensive understanding of the mechanisms of plant heavy metal tolerance.  Related Articles | Metrics

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Review for development of microbial diversity during dark tea fermentation period XU Zheng-gang, WU Liang, LIU Shi-quan,HUANG Hui-min,DONG Meng, ZHAO Yun-lin 2019, 36 (3):  92.  doi: 10.3969/j.issn.2095-1736.2019.03.092 Abstract ( 514 )   PDF   Save Dark tea, as a kind of microorganism fermented tea with important economic and health value, is closely related to microorganisms, and is involved in the productive process of dark tea. In order to clarify the relationship between microbial species and dark tea quality, we reviewed the variation of microbial diversity in the fermentation process. We compared the microbiological differences between different types of dark tea during fermentation period. We explained the changing rule of microorganisms in the pile-fermentation process of dark tea. Bacteria, Mycete and Saccharomycetes were widely propagated in the early, middle and late stage of piling fermentation, and the early stage of bacteria was more. Mycete and Saccharomycetes play dominant roles in the middle and late stage, respectively. By means of the clear advantages of modern identification of bacteria species composition and diversity in microbial taxonomic status of dark tea follow-up study, exploring effects of different microbial assemblages of dark tea quality, strengthening tea pile-fermentation control technology research and deep processing mechanization are important research trend in the future. According to the development experience of the world′s largest tea brand, Lipton Black Tea, we insist on ensuring quality and ensuring stable quality standards. According to uniform production standards, we produce and sell dark tea varieties from different origins in accordance with uniform standards, forming an internationally renowned brand. This is also a reference for the current crackdown on the development of the national dark tea industry. Related Articles | Metrics

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Study and policy proposals of the mechnism of IPR protection’s influence on the innovation models of crops transgenic technology WANG Yu, SHEN Wen-xing 2019, 36 (3):  96.  doi: 10.3969/j.issn.2095-1736.2019.03.096 Abstract ( 312 )   PDF   Save Intellectual property protection has become increasingly important during the R&D process of genetically modified crops. Its role has shifted from a defensive approach to a strategic tool, which has significantly affected innovation models of crops transgenic technology. Considering a certain order during the process of technology innovation which makes it a dynamic game, on the basis of Starkelberg Model and two key indicators of yields and profits, this paper analyzes the performance of original innovation and derivative innovation in the environment of industrial competition, and establishes models to devise formulas forinnovation performance calculation including yields, profit, etc. It concludes that intellectual property protection can directly affect innovation performance, thus affecting innovation models of crops transgenic technology based on case studies. Based on the strategic analysis of intellectual property protection, suggestions have been further brought up in this thesis to improve the policies of intellectual property protection on Crops Transgenic Technology. Related Articles | Metrics

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Modification of protease via click chemistry and its application SHAO Yi-wen, WANG Qiang, YU Yuan-yuan, WANG Ping, YUAN Jiu-gang, FAN Xue-rong 2019, 36 (3):  100.  doi: 10.3969/j.issn.2095-1736.2019.03.100 Abstract ( 387 )   PDF   Save Savinase was modified with dextran via click chemistry so as to enlarge the molecular weight, which can be applied for shrink resistance of wool. The structure changes of modified Savinase were assessed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), circular dichroism (CD) and fluorescence spectroscopy. Enzymatic properties and kinetic parameters were also determined. The results indicated that the molecular weight of the modified Savinase was significantly increased. After modification, the secondary structure of enzyme changed and the substrate affinity lowered, while the effects of temperature and pH on enzyme activities were similar. For the shrink-resistance finishing of wool, it could be found that the wool fabric treated by modified Savinase could meet the machine-washable standard with obviously decreased strength loss. Related Articles | Metrics

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The evaluation of virus enrichment methods for norovirus diversity in oysters GUO Ping, PAN Ying-jie, YU Yong-xin 2019, 36 (3):  104.  doi: 10.3969/j.issn.2095-1736.2019.03.104 Abstract ( 323 )   PDF   Save Oysters, the main vector of NoVs, contain almost all GI and most GII genogroup. In order to explore the diversity of the NoVs in oysters, this study aimed to evaluate four virus enrichment methods which were usually used in metagenomics study (PEG6000 precipitation method, ultracentrifugation method, filtration method and direct extraction method). Here, reverse transcription quantitative PCR (RT-qPCR) was adopted to calculate the Lentivirus (with GII.4 NoVs gene) copies before and after treated with the four methods, and recovery rate of Lentivirus was the evaluation indicator. Besides that, 16S rRNA PCR was conducted to compare the microorganism contamination. Both results showed that these four methods had no obvious difference, and the direct extraction method had relatively higher recycle rate and more serious microorganism pollution. It demonstrated direct extraction method has relatively good superiority, which is more suitable to extract RNA for NoVs diversity in oysters. Related Articles | Metrics

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The application of hop (Humulus lupulus) seedling sex determination by double fluorescence banding in the biology experimental course CHEN Chun-li, FENG Yan-ni, HUANG Tao 2019, 36 (3):  108.  doi: 10.3969/j.issn.2095-1736.2019.03.108 Abstract ( 314 )   PDF   Save Chromosome fluorescence banding technology not only can reveal chromosomal basic features including size, primary constriction, secondary constriction, satellite etc., but also contributes to karyotpe analysis. Obviously, it plays a significant role in biological studies, such as plant evolutionary biology, chromosome research in human and animal, and so on. However, there has been no chromosome fluorescence banding related to biological experimental teaching for undergraduates and graduates in colleges at present. Therefore, for students experimental practice, we used Humulus lupulus, an economic crop, as material and designed a complete experimental teaching program, containing chromosome preparation and fluorescence microscopy technology. The experiment shows the chromosome fluorescence banding technology visually and systematically, and the result is meaningful with potential application value. Meanwhile, student can learn lots of experiment skills and develop their knowledge during this experimental practice. Besides, this experimental teaching program provides some beneficial suggestions for popularizing chromosome fluorescence banding technology in biological teaching course. Related Articles | Metrics

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Practice and thinking of cell biology course teaching LI Yuan, LI Qiao-xia, YANG Ning 2019, 36 (3):  112.  doi: 10.3969/j.issn.2095-1736.2019.03.112 Abstract ( 386 )   PDF   Save Cell biology is a science that studies and reveals the basic law of cellular life activities, as an important basic and frontier discipline in modern life sciences as well, which has been one of the core courses of biology and biotechnology undergraduate. From the perspective of characteristics of the subject and taking the general higher education "11th Five-Year" national planning materials "Cell Biology" (fourth edition) written mainly by professor Zhai Zhonghe ,Wang Xizhong and Ding Mingxiao as an example, this paper sums up some specific problems and measures about teaching content arrangement and teaching methods improvement combined with years of teaching practice of the authors themselves to provide some teaching references so as to improve the teaching quality and learning effect of the course. Related Articles | Metrics

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Establishing a new experiment on skeletal muscle contraction in experimental course of physiology WANG Hui-ping, YAO Guo-hu, MA Ji-ying, ZHOU Xiao-hui, SUN Yi-ze, CHANG Hui 2019, 36 (3):  115.  doi: 10.3969/j.issn.2095-1736.2019.03.115 Abstract ( 336 )   PDF   Save The purpose of this study was to establish a new experimental methodology on skeletal muscle contraction based on own scientific research, and develop it into the experimental course of physiology. The experiment was carried out on partly dissociating soleus of simulated weightlessness and normal rats, and electrical stimulation was used to induce soleus to contract. Additionally muscle mass, contraction force, contraction time course, and the activity of citrate synthase in soleus were compared. The presented results reveal that not only the relationship between the skeletal muscle contraction with electrical stimulation form, but also the disuse atrophy of skeletal muscle resulted from simulated weightlessness. The new experiment is easily verified, understood and executed. It can achieve the instructional objectives of conventional skeletal muscle contraction experiment. Furthermore, it provides a chance for university students to understand the basic concepts of human′s health in space and develop their learning interests. Related Articles | Metrics

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Establishment and practice of assessment system in plant physiology experiment LI Yan-hua, JIN Ying-hong 2019, 36 (3):  118.  doi: 10.3969/j.issn.2095-1736.2019.03.118 Abstract ( 319 )   PDF   Save According to the characteristics of "plant physiology experiment" course, a set of scientific reasonable examination methods and grading standard of student′s academic record in plant physiology experiment course were set, which can promote the long-term development of this experiment teaching. This paper took the teaching practice as the starting points, evaluated students′ plant physiology experiments from five aspects: preview before class, experimental attitude, experimental operation, experimental reports and final examination. And finally, the good results were achieved. Related Articles | Metrics

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Discussion of teaching reform of hydrobiology for aquarium science and technology#br# GAO Xiao-chan, REN Hong-tao, XIONG Jian-li 2019, 36 (3):  121.  doi: 10.3969/j.issn.2095-1736.2019.03.121 Abstract ( 362 )   PDF   Save On the basis of analyzing problems existing in academic and practical teaching of hydrobiology, a series of teaching reforms，optimization of teaching content, careful preparing of courseware, improvement of traditional teaching mode, compilation of experimental teaching materials, construction of specimens library and instruments, improvement of the assessment methods included，were promoted. The purposes of mobilizing the enthusiasm of students, developing comprehensive ability, and improving hydrobiological teaching effect have been achieved. Related Articles | Metrics

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Practice and analysis the teaching effectiveness about genetic engineering promoted by professional competition DING Dong, LI Chang-zhong, ZHANG De-jun, HE Tao, DUAN Rui-jun 2019, 36 (3):  124.  doi: 10.3969/j.issn.2095-1736.2019.03.124 Abstract ( 342 )   PDF   Save With the aim of improving the theoretical teaching effectiveness of genetic engineering, the student-oriented teaching reform was proposed. The professional competition platform, which takes students as center, was introduced into teaching practice based on the educational program of college. As a result, the professional competition could provide a platform to display individual capabilities independently, develop the learning of theoretical knowledgement effectively, correct the research idea, cultivate the research ability, provide the platform to communicate, and optimize the learning atmosphere. Related Articles | Metrics

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Study on the practice teaching based on cases of wildlife trace evidence in forensic center of forest police HUANG Ya-lin, XU Yan-hong, HU Jing-lin 2019, 36 (3):  127.  doi: 10.3969/j.issn.2095-1736.2019.03.127 Abstract ( 352 )   PDF   Save The course of "biological trace evidence test" is a practical interdisciplinary course. It integrates the relevant knowledge of inspecting and testing of animals and plants trace evidence into the traditional human biological evidence technology, including forensics, zoology, plant science, biochemistry, molecular biology, immunology and other multidisciplinary content. The course is mainly established for criminal technology specialty in forest police colleges and universities. Considering learners′ biological knowledge is lacking, it is difficult to improve students′ subjective initiative only by class lecturing. Therefore, combining the professional characteristics of forest public security, we make full use of a large amount of practical cases of wildlife trace evidence in forensic center of forest police located at our school. Through case studying, simulation on-site teaching and student-assistant testing to create a good heuristic and interactive practice teaching system, we help students to stimulate interest in the study of biological trace evidence to improve teaching effectiveness. Related Articles | Metrics