Current Biotechnology

Progress on Stress Resistance Genes of Foxtail Millet
ZHAO Jinfeng1, LIAN Lingyan2, YU Aili1, WANG Gaohong1, DU Yanwei1, LI Yanfang1, ZHANG Zheng1, ZHAO Genyou1
2018, 8(1): 1-6. DOI: 10.19586/j.2095-2341.2017.0049
Abstract ( ) PDF (1085KB)( )

The molecular mechanism of stress resistance in plants is one of research focus at present. The aim of research is to explain the mechanism of adaptation to adversity in plant, to obtain various kinds of stress resistance genes and to improve plant stress tolerance through genetic engineering. In this paper, the molecular mechanism of plant stress response was introduced briefly. The recent progress on stress in millet, well known for its drought and barren resistance, were also overviewed. Finally, the prospect of millet genes function research were prospected, which was aimed to provide new insight for new varieties breeding with high stress resistance efficiently.

Advances of Adipose-derived Stem Cells Enhancing the Healing of Tendon Injury
WANG Jingjing1,2, MA Yuehui1*, GUAN Weijun1*
2018, 8(1): 7-13. DOI: 10.19586/j.2095-2341.2017.0135
Abstract ( ) PDF (1131KB)( )

Adipose-derived stem cells (ADSCs) have more advantages than other tissue derived mesenchymal stem cells (MSCs), such as a wide range of sources, easy to acquire, amplification in vitro, as well as low immunogenicity, etc. More and more studies begin to pay close attention to how they can be applied to the treatment and regeneration of tissue damage. This paper reviewed the discovery and separation of ADSCs, and their differentiation ability towards tendon lineage, as well as the application in the aspect of tendon injuries both at home and abroad in recent years. The paper also analyzed the advantages and problems of ADSCs application. At last the future research direction was prospected.

Progress on Synergistic Mechanism of the Insecticidal Activity of Bacillius thuringiensis
LI Chaofeng
2018, 8(1): 14-20. DOI: 10.19586/j.2095-2341.2017.0067
Abstract ( ) PDF (1711KB)( )

In order to solve the problems existing in the application of Bacillius thuringiensis (Bt) preparations, such as poor readily availability, short duration action, unstable control effect and emerging drug resistance, various additives have been widely used in Bt biopesticide and achieved the synergistic effect on pests control. In this paper, the studies on synergistic mechanism and action pathways of various additives on Bacillus thuringiensis at home and abroad were reviewed, and the potential and means of their actions were also pointed out, which was expected to provide references for investigating the insecticidal mechanisms of Bacillus thuringiensis.

Progress on Molecular Modification and Biological Activities of Polysaccharides From Grifola frondosa
YU Ping, LI Qian, WANG Wei, ZHENG Daheng, ZHANG Weijie, ZHAO Ting, ZHANG Min, MAO Guanghua, FENG Weiwei, YANG Liuqing*
2018, 8(1): 21-27. DOI: 10.19586/j.2095-2341.2017.0112
Abstract ( ) PDF (1115KB)( )

Polysaccharides extracted from Grifola frondosa exhibit various bioactivities, such as immunostimulatory, antioxidant, antitumor and so on. Molecular modification is an important way to improve the original activities or develop new activities. In this paper, the molecular modification methods, biological activities and structure-activity correlation of modified Grifola frondosa polysaccharides were reviewed, which was expected to provided reference for the full development and further study of Grifola frondosa polysaccharides.

Progress on Detection Methods and Biological Functions of Glycation Antibody Drug
ZHANG Bin1, LIU Xiaozhi1, ZHOU Jinghua2, WEI Jingshuang1, GAO Jian1, WANG Zhiming1*
2018, 8(1): 28-33. DOI: 10.19586/j.2095-2341.2017.0042
Abstract ( ) PDF (1078KB)( )

Glycation is an important protein modification that could potentially affect bioactivity and molecular stability of the rapeutic proteins such as monoclonal antibodies. Many studies suggested that elevated levels of glycated hemoglobin were closely related with cardiovascular disease and atherosclerosis. Human plasma proteins, such as albumin, globulin, fibrinogen and collagen may be glycosylated, and then forms AGEs. In this paper, we reviewed the root cause and the current analytical methods of glycation of the rapeutic antibodies. We also summarized the effect of glycation on biological function, which was expected to provided reference for research on development, optimizion and storage condition of the rapeutic antibodies.

Analysis of Location and Expression Pattern of Maize Expansin Gene Family
JIANG Zhilei, ZHOU Lin, WU Fengci, SONG Xinyuan*
2018, 8(1): 34-40. DOI: 10.19586/j.2095-2341.2017.0030
Abstract ( ) PDF (3702KB)( )

Expansins are active proteins that relax the cell walls of plants, which play an important role in plant development. In this study, we used the genomic database of PLAZA, NCBI, MaizeGDB, Uniprot, PLEXdb to get gene sequences, chromosome location, protein sequences and lengths of the maize expansins, constructed the phylogenetic tree, and analyzed the expression pattern of these genes at different tissue. The results showed that a total of 93 Expansin genes were systematically identified from maize and located on 9 chromosomes, and most of maize expansins have 250~300 amino acids. Maize Expansin genes have 40 EXPA, 47 EXPB, 6 EXLA, but no EXLB found. Forty-four specific expression genes were found in different tissues. The results of this study not only laid a foundation for further study of maize Expansin genes, but also provided the method for other researchers to acquire the gene information they studied.

Equivalence Study of CryNGc Protein Expressed in Transgenic Maize and Escherichia coli
ZHAO Fangfang1, LIU Yang2, JING Legang1, YIN Yuejia2, FENG Shudan1*, LIU Xiangguo2*
2018, 8(1): 41-45. DOI: 10.19586/j.2095-2341.2017.0152
Abstract ( ) PDF (2740KB)( )

The important precondition of safety evaluation of protein in genetically engineered maize is protein identification and equivalence study of proteins expressed in genetically engineered maize and Escherichia coli. CryNGc protein was expressed in E. coli through optimized methods such as codon optimization, cryNGc insecticidal protein gene synthesis, prokaryotic expression vector construction and E. coli expression conditions optimizition. On this basis, the characteristics and consistency of the recombinant protein HiII-NGc-1 in maize and CryNGc expressed in E. coli were analyzed by SDS-PAGE, immunoblot analysis and liquid chromatography-tandem mass spectrometry. The results showed that there was consistent in the expression of cryNGc gene in transgenic maize and E. coli by electrophoresis mobility, immune competence and amino acid composition consistent. The results confirmed that the CryNGc protein characteristics of maize transformants HiII-NGc-1 was consistent with the theoretical prediction, and was substantially equivalent to the CryNGc protein expressed in E. coli. The results provided the theoretical basis and data support for further biosafety evaluation of the new insect-resistant recombinant protein CryNGc maize transformants.

Molecular Detection of Rice Blast Resistance Genes Pi-ta, Pi-b, Pi54 and Pikm in Japonica Rice of Huang-huai Region
CHEN Feng1§, XU Jiandi1§, JIANG Mingsong1, LIANG Shuimei2, ZHANG Quanfang2, WANG Wenliang3, LI Guangxian1, YANG Lianqun1,2, ZHU Wenyin1*, ZHOU Xuebiao1*
2018, 8(1): 46-54. DOI: 10.19586/j.2095-2341.2017.0115
Abstract ( ) PDF (2731KB)( )

In this study, thirty-two varieties of the middle late maturing group and machine transplanting group district experiments in Shandong Province as well as sixteen cultivars of rice breeding consortium district experiments of Lianyungang Agricultural Academy of Agricultural Sciences were detected with the functional markers of rice blast resistance genes Pi-ta, Pi-b, Pi54 and Pikm. Based on genotype and rice blast resistance identifications, correlation analysis between genotype and phenotype was conducted. Results showed fifteen varieties containing rice blast resistance gene Pi-ta, twenty-five varieties containing Pi-b, twenty-six varieties containing Pi54, and twenty-one varieties containing Pikm in fourty-eight varieties. Luzidao 7 and Lianjing 14ZD24 contain four resistance alleles. Thirteen varieties, including YS-6-6, Jidao 1 and D400, etc, contain three resistance alleles. Eight varieties, including Lindao 10, Feng dao 2, Tianhenuo 303, etc, have no resistance alleles. The results of rice blast identification indicated that four varieties,including Jidao 1,Shengdao 072, Lianjing 14JD24 showed moderate resistance (MR). Twenty-six varieties, including Lindao10, YS-6-6, Shengdao 053 showed moderate susceptibility (MS). Fifteen varieties, including Jindao180, Lin 13-105, Shengdao 504 showed susceptibility (S).H11-15,Runnong9, Jindao160 showed high susceptibility (HS). Pi-ta, Pi-b, Pi54 and Pikm, have been widely used in rice blast resistance breeding in Huang-huai region. Correlation analysis showed that the rice blast resistance gene Pikm was significantly correlated with blast resistance overall index (r=0.477 5，P<0.01）.

Improving Artemisinin Content of Artemisia annua L. Through Overexpression of Flavanone 3-hydroxylase Gene (AaF3H)
ZHANG Tingting1, MA Jiawei1, WANG Luyao1, TANG Kexuan1, LI Shan2*, ZHAO Jingya1*
2018, 8(1): 55-62. DOI: 10.19586/j.20952341.2017.0073
Abstract ( ) PDF (2680KB)( )

Flavanone 3-hydroxylase (F3H) is a crucial enzyme involved in the flavonoid biosynthesis. To study the effect of F3H on artemisinin in Artemisia annua L., AaF3H was cloned from Artemisia annua L. in this study. The completed open read frame of AaF3H was 1 095 bp, and it encoded 364 amino acids. Southern blot showed that it was a single copy gene in A. annua genome. The AaF3H transgenic plants were generated by transforming AaF3H overexpression vector into A. annua, and the artemisinin content of AaF3H transgenic A. annua was determined by HPLC. The results showed that overexpression of AaF3H significantly increased the content of artemisinin. As the key enzymes in artemisinin synthesis, the expression of amorpha-4,11-diene synthase gene (AaADS), amorpha-4,11-diene 12-hydroxylase gene (AaCYP71AV1) and artemisinic aldehyde Δ11(13) reductase gene (AaDBR2) was also increased in AaF3H-overexpression transgenic A. annua plants by real-time quantitative PCR analysis. These results demonstrated that overexpression of AaF3H gene was an effective method to improve the content of artemisinin in A. annua.

Screening and Functional Research of Broad Spectrum Antagonistic Strains Against Forest Pathogenic Fungi
LIU Caixia1，2, JIAO Ruzhen1*, ZHAO Jingjing1
2018, 8(1): 63-70. DOI: 10.19586/j.2095-2341.2017.0113
Abstract ( ) PDF (2843KB)( )

Broad-spectrum antagonistic bacterium against forest pathogenic fungi, such as Fusarium oxysporum Schltdl, Haematonectria haematococca (Berk. & Broome) Samuels & Rossman, Colletotrichum gloeosporiodes Penz, Pestalotiopsisapiculatus (Huang.) Huang, were screened by plate confrontation method and mycelial dry weight method from nitrogen fixing bacteria in soil. The ability of phosphorus solubilization and potassium dissolving of strains were also studied. The results showed that there were 24 strains had strong antagonistic ability to 4 pathogenic bacterium, 11 strains of dissolved inorganic phosphorus, 7 strains of dissolved organic phosphorus and 4 strains of releasing potassium. The results of strain identification showed that strain 2024 was Bacillus subtilis subsp. subtilis and the strain 7014 was Pantoea vagans.

Package of Recombinant Adeno-associated Virus Type 2 Based on BmNPV-silkworm Expression System
LU Nian1, LIU Xingjian1, HU Xiaoyuan1, LI Yinv1, YI Yongzhu2, ZHANG Zhifang1*
2018, 8(1): 71-77. DOI: 10.19586/j.2095-2341.2017.0047
Abstract ( ) PDF (3018KB)( )

Adeno-associated virus (AAV) is the most commonly employed viral vector for gene therapy. The AAV currently used in gene therapy is mainly packaged with Autographa californica nucleocapsid polyhedrosis virus-Spodoptera frugiperda cell 9 (AcMNPV-sf9 ) system, however，the high packaging cost in AcMNPV-sf9 system limits its widespread application. Due to the lower cost and higher packaging quantity of silkworm baculovirus expression system, in present study, silkworm baculovirus was employed to package recombinant AAV (rAAV). The AAV2 genes (cap and rep) were optimized and then cloned into baculovirus transfer vector pVL1393 separately. Utilizing enhanced green fluorescent protein (EGFP) gene and luciferase (Lus) gene as report genes, and cloned them into viral transfer vector pVL1393-ITRs-MCS, which contained cytomegalovirus-IE promoter. Then co-transfected them into BmN cell line with the replication-inducible BmBacmid virus DNA, and harvested recombinant BmNPV particles containing cap, rep and report genes. Afterwards, the purified recombinant viruses (reBm-Cap2, reBm-Rep2) were mixed with reBm-EGFP and reBm-Luc, respectively. Then silkworm larvae were infected by recombinant viruses, and collected their expression products to obtain and purify rAAV containing target genes. To determine successful packaging of rAAV2 in silkworm expression system, the expression of EGFP and Luc in mammalian cells was tested. The results revealed that rAAV2 was successfully packaged in silkworm baculovirus expression system, and the expression of the report genes was achieved in mammalian cells.

Canada Agricultural Biotechnology Annual (2016)
LIU Ying, WEN Xiaojie, CAI Jingjing*, YANG Wenhan(Compiler)
2018, 8(1): 78-89. DOI: 10.19586/j.20952341.2018.0006
Abstract ( ) PDF (1150KB)( )

Canadian planting of biotech crops is estimated at about 10.3 million hectares for 2016, flat from the previous year. The main biotech crops remain canola, corn and soybeans, with small amounts of sugar beets added recently. Canada is one of a few countries to approve stacked traits, or to allow for multiple genetically engineered traits in a plant. In 2016, the AquAdvantage salmon was the first genetically engineered animal to receive food and animal feed approval in Canada.

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