Biotechnology Bulletin

Analysis of Mutation Sites of OsBADH2 Gene in Fragrant Rice and Development of Related Functional Marker

SUN Ping-yong, ZHANG Wu-han, SHU Fu, HE Qiang, ZHANG Li, PENG Zhi-rong, DENG Hua-feng

2021, 37(4): 1-7. doi:10.13560/j.cnki.biotech.bull.1985.2020-1086

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Fragrant rice is favored worldwide by consumers and rice breeders,due to its full aroma and high nutritional value. Exploring and utilizing the flagrant gene resources would lay a foundation for breeding excellent-quality varieties. Loss of function of the betaine aldehyde dehydrogenase gene（OsBADH2）leads to that rice is of fragrance. OsBADH2 coding regions in 10 fragrant rice varieties were sequenced to explicit its mutant type. A functional marker E7 was designed according to the 8-bp deletion in exon 7 of OsBADH2 in the fragrant rice. Genotypes of 10 fragrant rice varieties and 2 populations derived from fragrant rice/non-fragrant varieties were identified and verified by E7. The results indicated that 8 bp deletion and 3 bp replacement in the exon 7 of OsBADH2 gene were found in 9 cultivars,including Xiangyaxiangzhan,Huaxiang B and Nongxiang 24 et a1. And 1 bp（G）insertion mutation in exon 14 in the fragrant rice of Xiao7 appeared. The homozygous dominant,homozygous recessive and heterozygous genotypes of OsBADH2 could be rapidly and accurately distinguished by functional marker E7.

Expression,Protein Interactions and Biological Function Analysis of FaFT in Festuca arundinacea

LUO Wei, MU Qiong, SHU Jian-hong, WU Jia-hai, WANG Xiao-li

2021, 37(4): 8-17. doi:10.13560/j.cnki.biotech.bull.1985.2020-0927

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FLOWERING LOCUS T（FT）plays a significant role in regulating plant flowering. In this study,the FaFT gene was cloned from the previous tall fescue’s transcriptome in order to explore the FaFT gene biological function. qRT-PCR analysis showed that the FaFT gene maintained a certain circadian rhythm and was regulated by the light cycle in different developmental stages under long-day,short-day,continuous light,continuous darkness,day-night inversion,and long-day light. Subcellular localization revealed that the FaFT protein was located in the nucleus. FD（FLOWERING LOCUS D）protein that may interact with FaFT was selected by yeast two-hybrid,and the authenticity of the interaction between FaFT and FD protein was further verified by the bimolecular fluorescence complementation（BiFC）method. Through the construction of overexpression vectors and functional compensation,it was found that FaFT gene promoted Arabidopsis early flowering under long-day condition. After functional compensation of FT gene,the flowering phenotype of Arabidopsis mutants was restored,and the mutant plants themselves showed no flowering phenomenon. The above results indicate that the biological function of the FaFT gene of F. arundinacea has been successfully verified.

Genome-wide Identification and Expression Analysis of CBL Gene Family in Glycyrrhiza uralensis

GAO Ling, WANG Fei, XIE Shuang-quan, CHEN Xi-feng, SHEN Hai-tao, LI Hong-bin

2021, 37(4): 18-27. doi:10.13560/j.cnki.biotech.bull.1985.2020-1128

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CBL（calcineurin B-like proteins）,as the typical Ca²⁺ sensor protein in plants,plays important role in plant stress response and development. In order to discover the characteristics of the CBL family members of Glycyrrhiza uralensis and to investigate their functions,10 CBL genes were identified from the G. uralensis genome using bioinformatics method,designated as GuCBL1-GuCBL10,with the locations on 10 different scaffolds. Amino acid sequence alignment indicated that all CBL family members contained 4 EF-motifs that bound to calcium ion at the C-terminus,and some CBL members included myristoylation sites and palmitoylation sites at the N-terminus,which might be related to the subcellular localization of CBL protein. The phylogenetic tree analysis showed that the CBL gene family can be divided into 4 subfamilies,and each of the subfamilies contained Arabidopsis and soybean CBL gene family members,indicating that they had high homology. In the upstream promoter sequences of the G. uralensis CBL genes,there existed some cis-elements that responded to abiotic stresses and plant hormones,implying that the CBL family genes might have different biological functions. Tissue-specific expression analysis showed that GuCBL was diversely expressed in different tissues,with the highest accumulation in roots. qRT-PCR detection results indicated that,the CBL family genes showed different induced expressions under diverse stress treatments of salt,oxidation,drought,high temperature,and low temperature,respectively,suggesting the close associations between the expressions of CBL family genes and the responses to abiotic stress. All these results provide a reference for in-depth understanding the regulatory mechanisms of CBL genes in response to abiotic stress and for elucidating their functions in secondary metabolite accumulation.

Effects of Habitat,Temperature and Exogenous Hormones on the Organ Development of Karelinia caspia

BI Yuan-kun, LI Li, ZHU Chuan-ying, WANG Yan-qin

2021, 37(4): 28-34. doi:10.13560/j.cnki.biotech.bull.1985.2020-1126

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The growth and development of flower organs in plants are closely related to temperature,and high temperature can cause flower organs abnormal and changes in hormone content in the plant. To explore the effects of different habitats and exogenous IAA on the floral organ development of Karelinia caspia,the floral organ size of K. caspia（Female style and male filaments）under 2 habitats of desert and artificial green space and at different concentrations of IAA as well as the changes of hormone content in the floral organ were measured. Results showed that the appropriate temperature（27-30℃）promoted the growth of K. caspia flowering organs in the bud stage,while the high temperature（38-40℃）promoted the elongation of K. caspia stigma during flowering,thus affecting the growth of flower organs until maturity. External application of IAA at a certain concentration promoted the growth of K. caspia flower organs. In the desert,the IAA promotion effect of 0.3 μmol/L was the most obvious,while that in the artificial green space was 0.1 μmol/L.

Effects of Seedling Time on Overwintering Rate and Yield of Winter Oil Rapeseeds（Brassica rapa）in Over-exploited Water Area of Hebei Province

LI He-ping, LI Ji-ming, LI Ai-guo, WU Jun-yan, SONG Cong-min, SHEN Yan-ping, YANG Li

2021, 37(4): 35-46. doi:10.13560/j.cnki.biotech.bull.1985.2020-0881

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Studying the adaptability of winter oil rapeseed in Hebei province would provide theoretical reference for the reasonable planting of winter rapeseed in the local area. Twelve varieties with different cold tolerance were introduced and different sowing times（Sep. 5,Sep. 11,Sep. 17,Sep. 23,Sep. 29,and Oct. 5）were set to provide technical support for the popularization and application of winter rapeseed in Hebei province. Results showed that 12 introduced varieties crawled in the field,and their growth points were under ground. All varieties grew to 10-leaf-stage if sowing before Sep. 29,and overwintering rate was > 97%,and yield were high. All varieties failed to reach the 10-leaf-stage if sowing after Sep. 29,with smaller seedlings and lower overwintering rate,among which the yield sowing on Oct. 5 significantly reduced. The correlation analysis demonstrated that the yield was positively correlated with overwintering rate and root diameter,and negatively correlated with sowing date. According to cluster analysis results,12 varieties can be divided into 2 major classes,class Ⅰincluding Tianyou178,Tianyou 142,Hengyou6,Longyou9,with high yield. These 4 varieties adapted to the climate condition of Hebei Hengshui and were suitable for local planting. Class Ⅱ included 8 varieties（Tianyou4,15QX60,Longyou12,16RTS309,Hengyou8,16QD367,and Longyou8）,in which Tianyou4,15QX60 and Longyou12 were a subclass,with lower yield than class,thus they can be used as a reference to local plant varieties. 16RTS309,Hengyou8,16QD367 were a subclass,of which yield was low,it was inferred that they were suitable for southern area of Hengshui. Longyou8 was a subclass,and its yield was the lowest. It was inferred that it was suitable for northern area of Hengshui according to its growth habit. Therefore,Tianyou 178,Tianyou 142,Hengyou 6 and Longyou 9 can be directly used as cultivated varieties at Hengshui. Tianyou 4,15QX60,Longyou 12,16RTS309,16QD367,and Longyou 8 can be considered to be introduced to the north of Hengshui for testing.

LUO Li-li, ZHANG Hao, YANG Mei-xin, WANG Yun-fei, XU Jing-sheng, XU Jin, YAO Qiang, FENG Jie

2021, 37(4): 47-55. doi:10.13560/j.cnki.biotech.bull.1985.2020-1514

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Pathogenicity variation in Fusarium graminearum from different locations to wheat under different temperature conditions was compared. In 2018 and 2019,diseased wheat heads were collected from the winter wheat ecological region of Huang and Huai River Valleys and spring wheat ecological region of the Northeast China,a total of 110 Fusarium strains were obtained after single-spore isolation. The EF-1α sequences of the isolated strains were used to identify the Fusarium species. The results demonstrated that F. graminearum was the predominant species causing wheat head blight in the two ecological regions. Ten F. graminearum strains from each ecological region were randomly selected and inoculation was conducted by single floret injection method in two locations of Langfang representing the high temperate region and Xining representing lower temperature region. Significant difference of pathogenicity was observed in Xining. The F. graminearum populations from the Northeast China spring wheat region showed significantly higher pathogenicity than the population from Huang and Huai River Valleys wheat region. However,the pathogenicity of the two populations was similar in Langfang site. This indicates that the F. graminearum population in the Northeast China spring wheat region has a higher adaptability at low temperature. Results of the study indicate that temperature influences the pathogenicity of F. graminearum populations,which is significant for studying the pathogenicity differentiation of F. graminearum and comprehensive control and prevention of wheat head blight.

Difference Analysis of Fungal Community Among Bulk Soil,Rhizosphere and Rhizomes of Ligusticum chuanxiong Hort.

ZHANG Shu-hua, FANG Qian, JIA Hong-mei, HAN Gui-qi, YAN Zhu-yun, HE Dong-mei

2021, 37(4): 56-69. doi:10.13560/j.cnki.biotech.bull.1985.2020-0702

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It is aimed to obtain and compare the characteristics of fungal communities in bulk soil,rhizosphere and endophytic fungi of rhizome of Ligusticum chuanxiong Hort. High-throughput sequencing technology was used to analyze the structures and differences of fungal communities in bulk soil（S）,rhizosphere（R）and endophytic fungi（P）of the rhizome of L. chuanxiong Hort. FUNGuild was used to analyze the functions of fungal communities. There were high abundant fungal communities in the bulk soil,rhizosphere and endophytic fungi of the rhizome of L. chuanxiong Hort. The evenness and diversity of the fungal community showed as S>R>P,while the richness showed as R>S>P. Specific dominated fungal flora varied in the three groups at different taxonomic levels. Meanwhile,the fungal communities were mainly affected by the sample type,with the largest differences between bulk soil and rhizome,followed by that between rhizosphere and plants,and the least among the soils. Differences among origin areas were observed in the three groups separately,and ranked as S>R>P. Besides,the effect of environmental factors on the communities of the bulk soil and rhizosphere was S>R. The fungal community in the bulk soil was dominated by saprotroph,followed by pathotroph;while the rhizosphere soil was dominated by pathotroph,followed by saprotroph;and the endophytic fungi was dominated by pathotroph. The fungal structural characteristics and ecological functions presented continuous and gradual changes in the bulk soil to the rhizosphere soil and then to the rhizome of L. chuanxiong Hort.,which proved that the plants obtained special fungi from the soil during the cultivation process,continuous accumulation and reduction of these fungi occurred,and then special endophytic fungal community was formed. This way may be an important strategy for the construction of its microecological community structure and provide a scientific reference for understanding the formation of the authentic quality of Chinese medicine.

Effect of Lime on Preventing and Controlling Continuous Cropping Obstacle of Ganoderma lingzhi and Analysis of Its Microbial Community

YUAN Yuan, HUANG Hai-chen, LI Lin, LIU Guo-hui, FU Jun-sheng, WU Xiao-ping

2021, 37(4): 70-84. doi:10.13560/j.cnki.biotech.bull.1985.2020-0786

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The effects of lime treatment on the emergence rate of Ganoderma lingzhi,yield,disease rate and microbial community of G. lingzhi were revealed,which may provide theoretical basis for the prevention and control measures of G. lingzhi continuous cropping obstacles. Taking 4-years continuous cropping soil of G. lingzhi as control（CK）,the soil respectively was treated with 22.5 kg lime soaked（LI45）,45 kg lime soaked（LI90）and 45 kg lime scattered（LS90）,then Illumina NovaSeq platform was used to sequence the ITS1 and 16S rDNA amplicons of group CK,LI45 and LI90 respectively. The results showed that the lime soaking treatment at the same dose significantly improved the continuous cropping obstacles of G. lingzhi than the lime scattering treatment,and the change rates of G. lingzhi emergence,yield and abundance of partial microbial communities in the lime soaking treatment were dose-dependent. The effect in the LI90 was the most significant while compared with the CK,the rate of G. lingzhi emergence approximately increased by 68.89%,the yield approximately increased by 135.87%,and the disease rate approximately decreased by 54.90%. Comparing with the CK,the relative abundance of Ascomycota Aspergillus in the LI90 significantly reduced at the fungal genus level. At the bacterial genus level,the relative abundance of Proteobacteria Cupriavidus and Acidovorax were significantly higher in the soil of LI90 than that of the CK,and the relative abundances of Cupriavidus and Acidovorax increased in a doses-dependent manner. In conclusion,the continuous cropping barrier effect of G. lingzhi can be reduced by LI90 treatment,which may be related to the reduction of pathogens such as Aspergillus in continuous cropping soil,and the increase of various beneficial bacteria such as Cupriavidus.

Screening,Identification,Condition Optimization and Application of Efficient Phosphate Solubilizing Bacteria in Sandy Fluvo Aquic Soil

WEI Chang, QI Xiu-xiu, WU Yue, LIU Xiao-dan, WANG Yi, JIANG Ying, LIU Hai-tao

2021, 37(4): 85-95. doi:10.13560/j.cnki.biotech.bull.1985.2020-0550

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The objectives of this work are to isolate an efficient phosphate solubilizing bacterium from the sandy fluvo aquic soil,to study its effect on the change of soil phosphorus form and the growth of wheat,and to provide the theoretical basis for its application in practical production. PKO medium was used to isolate several phosphate solubilizing bacteria from sandy fluvo aquic soil,and their phosphate solubilizing abilities were compared to screen out an efficient phosphate solubilizing bacterium. The species of the bacterium was identified by combining the morphological,physiological and biochemical characteristics of the bacteria as well as 16S rDNA molecular analysis. The phosphate solubilizing conditions were optimized,and phosphorus-dissolving process was explored and growth-promoting effect of the bacterium in the soil was verified by pot experiment. As results,an efficient phosphate solubilizing bacterium W6 was screened and identified as Bacillus flexus. The results of shake flask experiment showed that the phosphorus dissolving capacity of the strain reached the maximum when the culture time was 24 hours,the initial pH was 6.0,the liquid content was 75 mL/250 mL,the carbon source was xylose,and the nitrogen source was potassium nitrate. The results from wheat pot experiment demonstrated that the contents of H₂O-P,NaHCO₃-Po,NaHCO₃-Pi,NaOH-Po,and NaOH-Pi increased by 127.3%,80.0%,54.5%,11.8%,20.0%,respectively,while the contents of hard-soluble HCl-P,concentrated HCl-Po,concentrated HCl-Pi,and concentrated H₂SO₄-P decreased by 10.9%,23.0%,42.1%,33.3%,respectively,when compared with the control. The root length,root surface area,root volume,root tip number and root diameter of wheat inoculated with the strain W6 increased 66.8%,57.0%,50.0%,13.6% and 9.1%,respectively,and the dry weight,nitrogen,phosphorus and potassium contents of wheat increased 70.5%,32.1%,24.7%,94.2% and 34.4%,respectively. In conclusion,the strain W6 may promote the transformation of phosphorus form from insoluble to effective,also promote the growth and development of wheat root system,improve the absorption and utilization efficiency of phosphorus by plants,and thus presents great application potential in agricultural production.

Study on Enzyme Production of a Chitinase-producing Strain Achromobacter sp. ZWW8 by Fermentation and Its Enzymatic Characterization

ZHANG Yao-xin, WANG Liang-jie, ZHENG Wen, XU Han-qin, ZHENG Lian, ZHONG Jing

2021, 37(4): 96-106. doi:10.13560/j.cnki.biotech.bull.1985.2020-0887

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A extracellular chitinase-secreting bacterium ZWW8 was isolated from shrimp shell-accumulated soil in Hubei and identified as Achromobacter spp. via 16S rDNA sequence analysis. The fermentation conditions for chitinase production by the strain ZWW8 were optimized using the single-factor experiment,and the optimal fermentation conditions were as follows：0.5% colloidal chitin,0.5% ammonium chloride,initial medium pH 7.0,and 37℃ for 2 d. Under the optimized fermentation conditions,the highest chitinase activity in the supernatant reached 33.9 U/mL,which was 8 times of the activity before optimization. By detecting the enzymatic properties of extracellular chitinase secreted by ZWW8 strain,it was revealed that the optimal temperature of chitinase produced by Achromobacter sp. ZWW8 was 50℃ and the optimal pH was 5. Ni ²⁺and Ca ²⁺ increased chitinase activity,but Hg ²⁺,Cu ²⁺ and Zn ²⁺ significantly inhibited the enzymatic activity. Meanwhile,the chitinase not only exhibited activity and stability at high salt concentrations,but also showed chitin binding capacity. The extracellular chitinase from the fermented broth purified by chitin resin column demonstrated high activity after urea removed by dialysis. The chitinase by Achromobacter sp. ZWW8 possesses excellent enzymatic properties,making it a highly efficient and stable biocatalyst,which lays a foundation for exploring its application potential in industry,agriculture and medicine,as well as the development and utilization of chitin resources.

Screening and Identification of Highly Efficient Crude Oil-degrading Bacteria and Construction of Bacterial Consortium

ZHU Shu-fang, EHENEDEN Iyobosa, NING Hai-jun, SHANG Jie-hao, LI Wu-yang, MENG Xian-gang

2021, 37(4): 107-115. doi:10.13560/j.cnki.biotech.bull.1985.2020-1094

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In order to obtain efficient crude oil-degrading strains,9 strains of microorganisms were isolated and screened from the oily sludge of an oil field in Shandong,laboratory’s own bacterial agents and crude oil contaminated soil,which were numbered SF1,SF2,SF3,SF4,SF5,SF6,SF7,SF8 and SF9. Studies have shown that 9 strains presented certain abilities of degrading crude oil. Via 16S rRNA identification,SF1 belonged to Ochrobactrum intermedium,SF2 and SF8 belonged to Enterobacter sp,SF3 and SF7 belonged to Acinetobacter sp,SF4,SF5,SF6 and SF9 belonged to Pseudomonas sp. Among them,strain SF1 has the strongest degradability,with a degradation rate of 48.73%,strains SF2,SF7,and SF9 showed strong degradability,with degradation rates of 41.90%,45.35%,and 32.25%,respectively. The degradation rates of other strains were higher than 20%. Through orthogonal experiments,it is concluded that the bacterial flora composed of strains SF1,SF2,SF3,SF4,SF5,SF6,SF7and SF9 with the addition amount of 2∶1∶1∶1∶1∶1∶2∶2 has an effect on crude oil,and the degradation effect is better than that by a single strain,and this flora is the best efficient one,with a degradation rate of 57.88%.

Construction of Mixed Fermentation System of Four PGPR Strains and Evaluation of Its Promoting Effect

ZHOU Jing, HUANG Wen-mao, QIN Li-jun, HAN Li-zhen

2021, 37(4): 116-126. doi:10.13560/j.cnki.biotech.bull.1985.2020-0989

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This objective of this work is to construct a mixed fermentation system from 4 plant growth-promoting rhizobacteria（PGPR）strains and to inoculate pepper for laying a foundation for the research and development of microbial compound agents. The medium and fermentation conditions of the mixed fermentation system were optimized by single factor experiment,orthogonal experiment and response surface method. And experiment of planting pepper in pot was conducted to evaluate the growth-promoting effect of compound broth. In mixed fermentation system,the optimal carbon source was sucrose,the optimal nitrogen source was yeast extract,and the optimal carbon-nitrogen ratio was 5∶1. The optimal fermentation conditions were as follows,temperature 29.7℃,pH 7.05,inoculation proportion 3.19%,and the medium volume 50 mL/250 mL. Under these culture conditions,the viable bacterial count in the mixed fermentation culture was 2.94×10¹⁴CFU/mL,which was about 5.13 times compared with single strain fermentation. The growth and physiological indexes of pepper seedlings were significantly improved after irrigating with the mixed fermentation broth,i.e.,the plant height,root length,fresh weight,and chlorophyll content increased by 8.01%,50.22%,43.66% and 6.94% compared with the single strain fermentation compound. In Conclusion,the compound agents prepared by the mixed fermentation system of 4 PGPR strains may have higher viable microbial counts,and play better growth-promoting effects on pepper,and have obvious application potential in the development of PGPR compound agent.

Polyhydroxybutyrate Production from Mixed Waste Cooking Oil by Cupriavidus necator

PAN Lan-jia, LI Jie, LIN Qing-huai, WANG Yin

2021, 37(4): 127-136. doi:10.13560/j.cnki.biotech.bull.1985.2020-1107

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Waste cooking oil,a kind of hydrocarbon with high carbon content,is one of the main valuable products from the disposal process of kitchen waste. In this study,Cupriavidus necator was employed to synthesize degradable bio-plastics,i.e.,polyhydroxybutyrate（PHB）,with mixed waste cooking oil as sole carbon source. The operational conditions of the PHB synthesis from the waste cooking oil were optimized,and the properties of the synthesized PHB were studied. To lay a foundation for the industrialization of PHB in the future,the PHB yields under different modes of fermentation were explored and compared. The results showed that the maximum yields of the C. necator cell dry weight and PHB were 9.5 g/L and 7.6 g/L under the conditions of 30℃,150 r/min,and pH = 7.0 with amount of 3% waste oil and inoculation addition. Furthermore,the yield of PHB reached 8.25 g/L after fermentation for 96 h by batch culture in a 5 L bio-reactor. The average molecular mass of the synthesized PHB in this study was 30 kD,and the polydispersity index（PDI）was 1.44. The melting temperature of PHB was 175.7℃ and a decomposition temperature of it was 285.5 ^oC,which may meet the requirements of thermoplastic materials.

Study on Antibacterial Activity and Mechanism of Cordycepin

GAO Su, MA Jie-xin, LIU Jing-ju, ZHAO Guo-zhu

2021, 37(4): 137-144. doi:10.13560/j.cnki.biotech.bull.1985.2020-1169

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Cordycepin is the main active component of Cordyceps militaris and other food and health products,the purpose of the study is to further evaluate and analyze the main antibacterial activity and mechanism of cordycepin. The bacteriostatic spectrum and effect of cordycepin on six bacteria were determined by minimum inhibitory concentration,bacteriostatic zone test and electron microscopic observation,and the growth curve,intracellular ultraviolet absorption substances leakage and morphological changes of the tested bacteria were further determined to analyze the mechanism of cordycepin inhibiting cell growth and destroying cell structure. The results showed that cordycepin had obvious bacteriostatic effect on three G⁺bacteria in the range of solubility,but had no obvious bacteriostatic effect on three G^-bacteria. Growth assay and electron microscopic observation showed that the morphology of G⁺ bacteria was seriously damaged after cordycepin treatment,the intracellular ultraviolet absorption substances were leaked out,and the bacterial growth process was blocked. While the surface of some tested G^-bacteria was slightly damaged and accompanied by a small amount of content leaked,but the bacterial growth process was not affected. Cordycepin presents certain selectivity in bacteriostasis,and it is easier to destroy the wall membrane system of the tested G⁺ bacteria and play a bacteriostatic role,but has no significant effect on the overall growth process of the tested G^-bacteria.

Antimony Accumulation and Its Effect on Antioxidation System in Different Tissues of Danio rerio

XU Kun, YANG Ai-jiang, HU Xia, ZOU Hai-tao, LI Bin, LIU Ji

2021, 37(4): 145-154. doi:10.13560/j.cnki.biotech.bull.1985.2020-1060

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This study aimed to study the accumulation of antimony in zebrafish tissues and changes in various enzyme activities and contents under the stress of different concentration of antimony. Danio rerio were exposed to antimony solution（0,13.26,26.53,and 39.79 mg/L）for 14 d to determine the accumulation of antimony,the activity of superoxide dismutase（SOD）,catalase（CAT）,content of glutathione（GSH）and malondialdehyde（MDA）in each tissue. The results showed that the order of antimony accumulation in each tissue was as：Liver > Gill> Muscle > Brain and it gradually increased with the increase of antimony stress concentration. With the increased of antimony stress concentration,SOD activity in the liver,brain,gills and muscles presented different trends;CAT activity and GSH content both increased first and then decreased,which was as promotion at low concentration and inhibition at high concentration;MDA content tended gradually increasing. In conclusion,antimony can accumulate in different tissues of Danio rerio,activating antioxidant defense system and causing lipid peroxidation.

Extraction of Scutellaria baicalensis Polysaccharides and Its Antioxidant and Antitumor Activities

LI Ping, HU Jian-ran, SHI Bao-zhong, ZHAO Jing-lei

2021, 37(4): 155-163. doi:10.13560/j.cnki.biotech.bull.1985.2020-0764

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The optimal method of extracting Scutellaria baicalensis Georgi polysaccharides（SBP）from was selected after comparing the extraction rates of SBP by water extraction and alcohol precipitation method,microwave extraction method and ultrasonic extraction method. After the purification of SBP,the ability of SBP scavenging hydroxyl radical and superoxide anion and its total antioxidant capacity were tested by antioxidant assays in vitro. Effects of SBP on the proliferation of gastric cancer cell MGC80-3 was detected by MTT assay. Effects of SBP on the morphology of MGC80-3 cells were evaluated by morphological observation. Then the potential molecular mechanism was explored by Western blot. The results showed that the yield of SBP obtained by microwave extraction was the highest（7.26%）. SBP demonstrated obvious scavenging ability to hydroxyl radical and superoxide anion,and had ideal total antioxidant capacity. SBP effectively inhibited the proliferation of MGC80-3 cells and affected their cell morphologies;SBP at low concentration improved Akt and p-IκB α levels. IKK β and pro-caspase 3 levels,LC3 B/LC3 A ratio and cleave-caspase 3/pro-caspase 3 ratio were positively correlated with SBP concentration. Therefore,SBP presents strong antioxidation and antitumor activities,and has potential value in the development of antioxidants and antitumor drugs.

Commercialization Development Trend of Genetically Modified Maize and the Enlightenment

JIAO Yue, HAN Yu, YANG Qiao, HUANG Yao-hui, AN Ji-cui, YANG Ya-zhou, YE Ji-ming

2021, 37(4): 164-176. doi:10.13560/j.cnki.biotech.bull.1985.2020-0803

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Based on the Report on the Global Status of Commercialized Biotech/GM Crops issued by the International Service for the Acquisition of Agri-biotech Applications（ISAAA）in recent 23 years,this paper made an in-depth analysis and elaboration on the commercialization process of global genetically modified maize from the aspects of planting area,application rate,approval status,research and development institutions,etc. Concurrently,this paper introduced and prospected the progress of the commercialization of genetically modified maize in China,aiming to provide policies and suggestions for the commercialization of genetically modified maize in China.

Progress in the Study of Allelopathy in Plants：A Case Study of Inhibiting Microcystis aerugingosa Growth

QIAN Kai-rong, MA Zeng-ling, LI Ren-hui, CHEN Bin-bin, WANG Min, ZHU Shu-nan, RONG Meng-wei, QIN Wen-li

2021, 37(4): 177-193. doi:10.13560/j.cnki.biotech.bull.1985.2020-1158

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Algal bloom caused by water eutrophication has still been a serious issue for the world now. Allelopathic substances released by plants have become an efficient and low-consumption control method of algal bloom,which is widely favored by domestic and international scholars. First,the effects of different aquatic plants and terrestrial plants on Microcystis aeruginosa growth are reviewed. Then,plant species,research methods,allelopathy effects,the allelopathy mechanism,and allelopathy characteristics are summarized. Further,the issues existing in the study of phytoallelopathic inhibition of algae are put forward. Finally,the development direction and prospect of plant algal inhibition effect are discussed.

Research Progress on Production,Scavenging and Signal Transduction of ROS Under Drought Stress

YANG Li, WANG Bo, LI Wen-jiao, WANG Xing-jun, ZHAO Shu-zhen

2021, 37(4): 194-203. doi:10.13560/j.cnki.biotech.bull.1985.2020-0804

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Drought stress is one of the major abiotic stresses and seriously hinders the production of crops. The increased level of ROS under drought stress causes oxidative stress,destroys lipid,nucleic acid and protein,and finally leads to cell damaged and even death. Therefore,it is critical to maintain the physiological level of ROS for aerobic organisms,through the coordinated control of enzymatic and non-enzymatic antioxidants. This article summarizes the production,scavenging and signal transduction of ROS under drought stress,which may provide a reference for studies on plant response to drought stress.

Research Progress of Floral Organ Vacuole pH Regulating Flower Color Formation

ZHANG Gai-tian, QI Hui, YANG Suo-ning, CHU Zhi-yun, TIAN Tian, YUAN Su-xia, LIU Chun

2021, 37(4): 204-210. doi:10.13560/j.cnki.biotech.bull.1985.2020-0615

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Flower color is an important character in ornamental plants. The formation and the regulation of flower color are affected by many factors. Vacuole pH value is one of the most important factors. This paper expounded the effects of vacuole pH in floral organ on the color of flower from chemical mechanism of the vacuole pH regulating color change,and the mechanism of proton pump regulating the pH change of vacuole. Additionally,the paper analyzed the existing key issues and gave prospects,which lays a foundation for further research in vacuole pH related genes function and molecular regulating mechanism,as well as is of significance for the color regulation and molecular breeding in ornamental plants.

Research Progress of the Effects of Glyphosate on Microorganisms

LIANG Xin, ZHANG Bao-shan, LIU Ji-rui, YU Si, CHEN Si-yu

2021, 37(4): 211-223. doi:10.13560/j.cnki.biotech.bull.1985.2020-0658

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Glyphosate is the most widely used and most produced agricultural herbicide in the world. It has played an important role in reducing labor costs,improving agricultural production efficiency,and promoting the development of genetically modified crops. With the continuous accumulation of glyphosate in the environment,their residues have been found in more and more organisms. The action mechanism of glyphosate on plants and microorganisms is similar. Using microorganisms as a tool to study glyphosate-resistant genetically modified crops is currently a hot research topic. However,little attention is paid to glyphosate’s direct effects on the growth and reproduction of microorganisms as well as the evolution of flora. This article mainly reviews the action mechanism of glyphosate on microorganisms,the effects on soil and beneficial microorganisms outside the soil,and microorganisms that can degrade glyphosate. This has certain guiding significance for further understanding and safe application of glyphosate as well as reducing environmental pollution.

Research and Application Progress of Bacteriocin

TIAN Lu, WU Mi, GOU Jing-xuan, GONG Guo-li

2021, 37(4): 224-233. doi:10.13560/j.cnki.biotech.bull.1985.2020-0863

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As food safety issues are increasingly valued by the international community,the application of bacteriocins that target food spoilage/pathogenic bacteria has received great attention. As a safe natural preservative,bacteriocin can effectively resist food-borne pathogens through different modes of action,like pore formation and inhibition of cell-wall/nucleic acid/protein synthesis. However,emergence and spread of mutant strains resistant to bacteriocins is hampering food safety. More and more people have begun to know the phenomenon of food pathogens resistances to bacteriocins,and this is conducive for people to have the deep study of bacteriocin drug-resistance. This article reviews the classification,synthesis,antibacterial mechanism,and production methods of bacteriocins,introduces the mechanism of bacterial resistance to bacteriocin and the methods of reducing bacteriocin resistance to drug,and further clarifies the potential application of bacteriocin as a natural preservative in food preservation.

Research Status in the Production of Squalene by Thraustochytrids

LI Meng-fan, XIE Yun-xuan, XIE Ning-dong, ZHANG Ai-qing, WANG Guang-yi

2021, 37(4): 234-244. doi:10.13560/j.cnki.biotech.bull.1985.2020-1148

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Squalene is an important polyunsaturated triterpene substance,participates in multiple physiological processes in human body,and has the effect of enhancing immunity,delaying skin aging,and preventing the cancer. Under heterotrophic conditions,Thraustochytrids can grow rapidly and synthesize high content of squalene,thus there is the potential of developing industrial production of squalene. In this paper,the application potential of ecology and biotechnology of Thraustochytrids were outlined,focusing on the research status of squalene production by Thraustochytrids,and the metabolic pathways of squalene synthesis by Thraustochytrids were summarized. Then,five main factors affecting the production of squalene in the fermentation of Thraustochytrids were reviewed,and the regulatory effects of hormone substances on the production of squalene by Thraustochytrids were summarized. The aim of this review is to provide reference for the further research and development of squalene production by Thraustochytrids.

Research Progress on Drosophila Intestinal Stem Cells and Intestinal Microflora

JIN Qiu-xia, WANG Si-hong, JIN Li-hua

2021, 37(4): 245-250. doi:10.13560/j.cnki.biotech.bull.1985.2020-0875

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There are strong immune system,stem cells and micro ecosystem in the intestine of animals. The intestinal microflora plays an important role in maintaining the normal intestinal epithelium and immune function. Under normal conditions,the intestinal microflora of a host is in a state of micro-ecology equilibrium,and its symbiotic relationship is maintained. The change of intestinal microflora can cause metabolic diseases by affecting intestinal epithelial cells,and even change the behavior pattern of the host. The physiological structure,the mechanism of stem cell proliferation and differentiation,and the microhabitat of Drosophila’s gut are highly conserved with mammals. However,the structure of Drosophila intestinal microflora is far less complex than that of mammals. Therefore,Drosophila is an ideal model for studying the interaction between host and symbiotic bacteria. This paper will introduce and discuss the mechanism of the growth and differentiation of the symbiotic bacteria and stem cells in Drosophila’s gut,which is of great scientific significance to reveal the pathogenesis of human gut related diseases.

Research Progress on Biological Function Growth and Development Related to N6-methyladenosine in Mammals

XUE Xiang-lan, DING Yang-yang, LIU Yue, LI Xiao-bo, JIANG Lin, HE Xiao-hong, MA Yue-hui, ZHAO Qian-jun

2021, 37(4): 251-259. doi:10.13560/j.cnki.biotech.bull.1985.2020-1358

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Epigenetic modification of RNA N6-methyladenine（m⁶A）is the most extensive intermediate chemical modification in RNA,which exists commonly in living organisms,especially in mammals. The discovery of m⁶A modifying enzymes and binding proteins confirmed that RNA methylation modification is a dynamic reversible process,and the common m⁶A modifying enzymes mainly include methyltransferase and demethylase. Recent studies have successively found that m⁶A modification level and m⁶A distribution on RNA affect the processes of mRNA splicing,translation,stability,and exporting. RNA methylation-mediated epitranscriptomics is involved in many biological processes. This review summarizes the major m⁶A methylation modification of enzyme,the detection method and m⁶A biological functions in embryonic development,muscle development,as well as lipogenesis based on the latest m⁶A research,aiming to find new directions for exploring the functions and regulatory mechanisms of m⁶A methylation modification in important biological processes in animals.

Advances in Studies on Identification Methods and Molecular Biology of Drought Resistance in Sorghum

JIANG Bai-yang, BAI Wen-bin, ZHANG Jiang-hua, FAN Na, SHI Li-juan

2021, 37(4): 260-272. doi:10.13560/j.cnki.biotech.bull.1985.2020-0941

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Sorghum[Sorghum bicolor（L.）Moench]is an important food,feed and energy crop in the world. Because of its developed root system,strong water absorption ability,high water utilization rate and strong drought tolerance,it is widely planted in arid and semi-arid areas,which has important research value in the field of crop drought resistance research. Drought is the main stress factor affecting the growth and development of sorghum. The drought resistances among different varieties of sorghum vary largely. Screening sorghum varieties with strong drought resistance is of importantly theoretical and practical significance for promoting the process of drought resistance breeding of sorghum. Identification methods and identification indexes of sorghum resistances during different growth and development periods,as well as the evaluation methods of drought resistant germplasm resources of sorghum are introduced in this paper. The progress of molecular genetic research on sorghum is reviewed from three aspects：transcriptome analysis,drought resistance QTL localization and drought resistance gene mining,and the development direction of drought resistance of sorghum is prospected,aiming to provide reference for further exploring the molecular mechanism of drought resistance of sorghum and speeding up molecular genetic breeding of drought resistance of sorghum.

A Comparative Study of Methods for Extracting Protein from Sesame Residue

JIAO Xiao-bo, WANG Gao-yang, LI Jin-hong, WANG Xiao-yi, MENG Guo-qing, CUI Jian-dong

2021, 37(4): 273-281. doi:10.13560/j.cnki.biotech.bull.1985.2020-1130

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Sesame residue is the main by-product of sesame oil processing,and contains rich proteins. Currently in addition to a few of them are used as feed or fertilizer,most of them is discarded as waste,which causes environmental pollution and serious waste of resources. Therefore,it has a broad application prospect and important practical significance to extract protein from sesame residue for high value utilization. In this study,sesame protein was extracted from sesame residues by the method of deep eutectic solvent（DES）and alkaline extraction followed by acid precipitation,respectively. A novel DES method of extracting sesame protein formed by sodium acetate（CH₃COONa·3H₂O,SAT）and urea was established. The effects of extraction temperature,time,solid-liquid ratio,and extraction times on the extraction rate of protein were investigated. The optimal extraction conditions of sesame protein by SAT-urea system were as follows：temperature 75℃,extraction time 180 min,solid-liquid ratio 1∶50,extracting twice under the conditions of using ethanol to precipitate proteins. Under the optimized conditions,the total protein extraction rate was 71.41%,and the purity of sesame protein was 89.43%. Compared with alkaline extraction followed by acid precipitation,the sesame protein extracted by DES presented high purity;moreover,DES can be recycled and used without waste pollution. These results show that DES extraction method is a new green method of extracting plant protein. This study provides a new insight for the green extraction of plant protein.

Novel Coronavirus and Research Progress of Related Clinical Detection Methods

LI Jia-jun, ZHENG Xiao, SHENG Jie, XU Yao

2021, 37(4): 282-292. doi:10.13560/j.cnki.biotech.bull.1985.2020-0954

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The novel coronavirus（SARS-CoV-2）pneumonia（COVID-19）broke out in Wuhan at the end of 2019 and rapidly spread to the whole world. It is transmitted mainly through respiratory droplets and close contact with a strong infectious capacity,and some patients are asymptomatic but able to infect others. The diagnosis of COVID-19 is particularly critical for the prevention and control of the epidemic,early detection,early isolation and early treatment of patients. The detection of SARS-CoV-2 is mainly based on nucleic acid and specific antibody detection. This paper summarized the pathogen characteristics and development of detection methods of the SARS-CoV-2,and briefly introduced the principles and application status of detection technology,aiming to provide some helps for the accurate clinical diagnosis as well as epidemic prevention and control of COVID-19.

Recent Advances on Dynamic Regulation and Imaging Techniques of RNA Polymerase II

QIAN Hong-ping, CHEN Bo, LIN Jin-xing, CUI Ya-ning

2021, 37(4): 293-302. doi:10.13560/j.cnki.biotech.bull.1985.2020-1015

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RNA polymerase II（RNAP II or Pol II）plays a vital role in regulating the growth and development of organisms and responding to environmental changes. RNA polymerase II assembles and locates at transcription factory,where multiple genes are recruited for the formation of co-transcription,which is a highly dynamic and coordinated process. Although the critical functions of RNA polymerase II in eukaryotic gene transcription have been widely studied,the dynamic characteristics of plant RNAP II under different physiological conditions are still poorly understood. The development of single-molecule imaging and labeling techniques in living cell will contribute to improving our new understanding of the dynamic characteristics of plant RNAP II in vivo. In this review,we mainly expound the structural and functional characteristics of RNAP II,and introduce the dynamic regulation and transcription mechanism of RNAP II. Furthermore,we put the emphasis on summarizing the recent progresses in dynamic labeling and imaging techniques of RNAP II in living cells,aiming to provide theoretical basis for further studies on the dynamic regulation of RNAP II in plant.

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2021, 37(4): 303-303.

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