Table of Content

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• Current Biotechnology
  Volume 9 Issue 5 25 September 2019
• Application Progress of Omics in the Research of Anther Development Ⅰ: Transcriptomics
• ZHANG Zaibao1,2§, ZHAO Hai1§, HU Menghui1, DENG Lijun1, WANG Qi1, LI Jiuli1, YUAN Hongyu1,2
• 2019, 9(5):  433-439.  DOI: 10.19586/j.2095-2341.2019.0045
• Abstract ( ) PDF (1063KB)( )
Anther development plays an important role in the sexual reproduction of flowering plants. Normal anther and pollen development is directly associated with seed production and crop yield. Anther development is a strictly regulated biological process. Many key genes play regulatory roles in the development of anthers. Mutations in these genes can lead to male sterility. Previous studies on anther biology focused on the functional study of single genes, but lacked an overall understanding of transcriptional changes, protein changes, and metabolic changes during anther development. With the development of high-throughput sequencing technology and the innovation of analytical methods, a variety of omics techniques have been successfully applied in the study of anther development, such as transcriptomics, proteomics, metabolomics, which greatly complement the regulation network of anther development. And it has gradually led to the systematic exploration and analysis of the general laws of anther development, which has  greatly promoted the stereoscopic study of anther development. In this paper, we summarized the application and research progress of plant anther and pollen in transcriptomics, aiming to enrich the information in the process of plant reproductive development, and provide a large theoretical basis for breeding work.
• Application Progress of Omics in the Research of Anther Development Ⅱ: Proteomics and Metabonomics
• ZHANG Zaibao1,2§, ZHAO Hai1§, HU Menghui1, DENG Lijun1, WANG Qi1, LI Jiuli1, YUAN Hongyu1,2
• 2019, 9(5):  440-448.  DOI: 10.19586/j.2095-2341.2019.0081
• Abstract ( ) PDF (1094KB)( )
Since the launch of the Human Genome Project, with the rapid development of various sequencing and identification techniques, people have obtained the  information  of sequence, expression, modification or metabolic for genome, transcriptome, proteome and metabolome in many species. At present, researchers have realized that it is difficult for any gene or protein to function independently. The interaction between genes or proteins is the basic way for them to regulate or participate in life activities. In addition, anther development is one of the two most important processes in plant sexual reproduction, and the successful development of various developmental events related to pollen development and release is the key to the success of sexual reproduction. At present, with the rise of omics technology, various omics techniques have also been widely used in the research of anther development. Among them, from the genetic central dogma, protein and many small biomolecules are the direct bearers of life activities. Therefore, large-scale in-depth study of plant anthers by proteomics and metabolomics will further expand and deepen people's understanding and application of anther development mechanism. In this paper, we summarized the progress of proteomics and metabolomics of plant anthers and pollens, and elaborated the application value and characteristics of proteomics in different research fields, providing a useful theoretical basis for related basic and applied research.
• Progress on  Ethylene Signal Transduction Pathway and Related Genes in Plants
• SHI Qingling1, LI Zhongfeng2, DONG Yongbin2*, LI Yuling2
• 2019, 9(5):  449-454.  DOI: 10.19586/j.2095-2341.2019.0080
• Abstract ( ) PDF (1578KB)( )
Ethylene is one of the important endogenous hormones in plants. Ethylene signal transduction pathway plays an important role in regulating plant growth, development, as well as biological and abiotic stress. This paper reviewed ACC synthase which is a key enzyme in plant ethylene synthesis, and key genes which play key roles in plant ethylene signal transduction pathway, such as ethylene receptor gene, negative regulatory factor CTR1, and transcription factor EIN2, EIN3/EIL1 and ERFs,  which was expected to provide reference for further research.
• Advances on Roles of Guard Cell Wall in Stomatal Movement
• ZHOU Ying1,2, ZHENG Lin2, WEI Jianhua2, MA Yan3, DAI Shaojun1*, WANG Hongzhi2*
• 2019, 9(5):  455-460.  DOI: 10.19586/j.2095-2341.2019.0047
• Abstract ( ) PDF (1136KB)( )
Stomatal guard cells play important roles in plant development by controlling water and CO2 exchange between the plant and the environment. The repeatable movements of stomata depend on the characteristics of guard cell wall. Biochemical and molecular studies with the help of computational modeling have greatly increased the understanding of the specific roles of guard cell wall components and characteristics on stomatal function. We summerized the recent advances on the essential roles of polarized wall of guard cell on the stomatal function, and the deposition process and evolutionary conservation of the polarized wall. Meanwhile, the latest finding of the specific influence of cell wall components on stomatal movement were also summarized, particularly in the fields of molecular genetics and biochemistry. We also proposed the future research on the molecular mechanism by which guard cell wall is involved in the function of stomata, which was aimed to provide new insight for genetic engineering of guard cell wall.
• Progress on Microbial Fermentation of Medicinal and Edible Plants
• LI Yubang1, WU Junlin1,2*, LI Mansha1
• 2019, 9(5):  461-466.  DOI: 10.19586/j.2095-2341.2018.0127
• Abstract ( ) PDF (1028KB)( )
Medicinal and edible plants refer to plants that can be used as both food and medicine, and they are widely used in daily health care. The medicinal value of medicinal and edible plants can be improved by applying the decomposition ability of microorganisms. The common strains, mechanism and characteristics, technical progress of medicinal and edible plants treated by microbial fermentation were reviewed, as well as their application in food and medicine. And the development trend was prospected in order to provide basis for the development of medicinal and edible plant products treated by microbial fermentation.
• The Regulatory Mechanism of Mitophagy and its Role in Related Diseases
• LIN Jingjing, YANG Yufeng*
• 2019, 9(5):  467-475.  DOI: 10.19586/j.2095-2341.2019.0065
• Abstract ( ) PDF (2480KB)( )
Mitophagy refers to the process of specifically removing damaged or redundant mitochondria, and it is an important mitochondrial quality control mechanism. Dysfunction or excessive activation of mitophagy will destroy mitochondrial homeostasis, affect body health and even lead to death. The mechanisms of positive regulation of mitophagy found in yeast and mammalian cells were mainly discussed. In yeast, mitophagy was mediated by autophagy-related protein 32 (Atg32), while there were mainly three regulatory pathways of mitophagy in mammals: PTEN-induced kinase 1(PTEN-induced kinase 1, PINK1)/E3 ubiquitin ligase Parkin pathway, NIP3-like protein X(NIP3-like protein X, Nix) pathway and FUN14 domain-containing protein 1 (UNDC1) pathway. In addition, several newly discovered mitophagy receptors could also mediate mitochondrial specific clearance. Moreover, the mechanisms of negative regulation of mitophagy, which were seldom studied at present, were reviewed. Finally, the relationship between abnormal mitophagy and human diseases (such as Parkinsons disease) was discussed. The molecular mechanism of the occurrence of mitophagy was in-depth analyzed in order  to provide theoretical basis for further research on the treatment of diseases related to abnormal mitophagy.
• Mechanism of Ca2+ in Cadmium Induced Nephrotoxicity
• REN Zhen1, ZHAO Jinfeng1, ZHANG Yao1, XIE Yimin2, ZHOU Yang1, GU Jie1*, SHI Haifeng1*
• 2019, 9(5):  476-482.  DOI: 10.19586/j.2095-2341.2019.0040
• Abstract ( ) PDF (1079KB)( )
Cadmium (Cd) as a toxic heavy metal, and the situation of pollution is becoming more and more serious in China. Cd accumulates in kidney after exposure to human body. In the kidney, Cd mainly accumulates in the renal tubules. Due to the long half-life of Cd in human body, Cd will cause the loss or disorder of renal function, and even increase the incidence of renal cancer. At present,  studies have suggested that Cd causes endoplasmic reticulum stress, calcium homeostasis disruption, and eventually leads to cell apoptosis. Ca2+ homeostasis plays a key role in maintaining the normal physiological function of the cells. Recently, the methods and drugs to restore Cd disrupted Ca2+ homeostasis has been tested. Further studies of the roles and the underlying mechanism of Ca2+ in Cd induced nephrotoxicity are urgently needed. This article reviewed the underlying mechanism of Ca2+ in Cd-induced nephrotoxicity, which might provide a new direction for the study of the mechanism of toxicity and detoxification in Cd.
• Study on Antagonizing Regulation of Shoot Growth by Abscisic Acid and Gibberellic Acid in Rice
• QI Yidong, QIN Hua, GAO Yadi, WANG Fangfang, HUANG Rongfeng, QUAN Ruidang*
• 2019, 9(5):  483-489.  DOI: 10.19586/j.2095-2341.2019.0042
• Abstract ( ) PDF (7959KB)( )
Abscisic acid (ABA) and gibberellic acid (GA) are both important plant hormones, which have important regulatory effects on rice growth. Previous studies have showed that the down-regulation of OsPK1 gene (encoding cytoplasmic pyruvate kinase) in rice results in the change of ABA/GA ratio, thus affecting the plant height. However, the molecular mechanism of shoot growth regulated by ABA and GA synergistically is still unclear. In order to reveal the relationship between ABA and GA in the regulation of shoot growth of rice seedlings, rice wild-type Hwayoung (HY) and IR29 were used as research materials, treated with plant hormones ABA, GA3 and GA synthesis inhibitor paclobutrazol (PAC), and shoot length and fresh weight were used as indicators to measure the growth and development of rice seedlings. The results showed that the inhibition of ABA on shoot growth of rice was relieved under gibberellin treatment. Subsequently, real-time fluorescence-based quantitative PCR (qPCR) was further used to detect the expression levels of key gibberellin synthesis genes (OsCPS1, OsKS1, OsKO2, OsKAO, OsGA20ox2 and OsSHB) in ABA-treated wild-type Nipponbare, and the results showed that ABA could inhibit the expression of OsCPS1, OsKO2 and OsSHB. The results have revealed that ABA could affect the expression of genes related to gibberellin biosynthesis through signal transduction, thus inhibiting the shoot growth of seedlings.
• Genome-wide Identification and Expression Pattern Analysis of Class Ⅲ Peroxidase Family in Gossypium raimondii
• GAO Zhengyin, SUN Wenjie, SONG Xiaoyun, HU Shi, ZUO Kaijing*
• 2019, 9(5):  490-501.  DOI: 10.19586/j.2095-2341.2019.0043
• Abstract ( ) PDF (9584KB)( )
Class Ⅲ peroxidases in plants, as the unique secretory peroxidases, are widely exist in extracellular space or vacuole, and participate in developmental and biochemical pathways in response to biotic and abiotic stresses. To analyse their functions in cotton fiber development, we searched Class Ⅲ peroxidases in Gossypium raimondii genome using GO Annotation and Conserved Domain Analysis. The results showed that G. raimondii genome contains 90 peroxidase genes, which can be classified into 8 subgroups. Transcriptome analysis revealed that the Class Ⅲ GrPrx genes have differential expression patterns in different tissues and different fiber developmental stages, in which GrPrx11 and GrPrx16 highly express in the secondary cell wall synthesis stage, indicating that the importance of Class Ⅲ GrPrxs genes during fiber development.
• Expression of Ovis aries λ3 Interferon in Silkworm-baculovirus Expression System and the Antiviral Activity Assay
• WANG Xianxiang1,2, ZHAO Ze2, WANG Peng2, LIU Xingjian2, HU Xiaoyuan2, ZHANG Zhifang2, LI Yinv2, FANG Lingli1, YE Aihua1*
• 2019, 9(5):  502-508.  DOI: 10.19586/j.2095-2341.2019.0036
• Abstract ( ) PDF (2536KB)( )
IFN-λ as a pleiotropic cytokine belongs to type-Ⅲ interferons and demonstrate fundamental role against viruses that escape from the host immune system. In order to efficiently express Ovis aries λ3 interferon (OvIFN-λ3) with high antiviral activity by using Bombyx mori baculovirus expression system, firstly OvIFN-λ3 gene was optimized and synthesized, then cloned into transfer vector pVL1393 to obtain recombinant plasmid pVL1393-OvIFNλ3, after that, the recombinant plasmid and inactivated-rescued Bombyx mori baculovirus shuttle vector reBmBac DNA were co-transfected into Bombyx mori ovarial passage cell Bm-N to obtain recombinant baculovirus containing OvIFN-λ3 gene. The recombinant baculovirus was then infected with the 5th instar silkworm larvae and the hemolymph of the silkworms was collected after the onset of the disease. The anti-virus experiment of recombinant vesicular stomatitis virus expressing green fluorescent protein (VSV-GFP) was carried out on sheep kidney cells by micro cytopathic inhibition method, and initial antiviral activity of OvIFN-λ3 was determined to be (6.5±0.27)×105 U/mL. And plaque assay was performed to purify the recombinant baculovirus and surprisingly it was demonstrated that antiviral potential of purified recombinant virus was (3.1±0.42)×106 U/mL, it indicated that the expression level was increased. The silkworm-baculovirus expression system provided a new method for mass-scale production and application of Ovis aries λ3 interferon products.
• Genetic Diversity and Phylogenetic Analysis on mtDNA Cytb in Yak of Karakorum-Pamir Region
• LI Jing1, WANG Li1, DAO Min2, HU Ping1, XIAO Yi1, HAN Jianlin1, WANG Yutao1*
• 2019, 9(5):  509-517.  DOI: 10.19586/j.2095-2341.2019.0060
• Abstract ( ) PDF (4788KB)( )
Yak is an important breeding resource in the Karakorum-Pamir Region, and there are few reports on the genetic diversity and phylogenetic evolution of yak in this region. In this study, the genetic diversity of mtDNA Cytb gene in yak in Karakoram-Pamir Region was analyzed. The results showed that the full length of mtDNA Cytb gene was 1 140 bp, and the average contents of T, C, A and G bases were 26.2%, 29.0%, 31.7%, and 13.1%, respectively, and the A+T base content (57.9%) was greater than C+G base content (42.1%), presence of base Basis bias; a total of 10 polymorphic loci with 5 haplotypes were detected, and only the transformed variant type was obtained; the average haplotype diversity, nucleotide difference, and nucleoside acid diversity were 0.599±0.001 23, 3.363 and 0.002 59, respectively. The results based on the construction of phylogenetic trees in the domestic yak population which indicated that the yak in the Karakoram-Pamir Region is closely related to the wild yak, and has a close relationship with the Bazhou yak; using the representative sequence of yak and bovine subfamily submitted by GenBank, the phylogenetic tree was constructed by the maximum likelihood method. The results showed that the yak was closely related to the American bison and had a close relationship with the common cattle.
• Study of Function of Rhamnose Metabolism Related Gene LRA3 and Cis-acting Elements in its Promoter PLRA3 in Pichia pastoris GS115
• LIANG Mingli, LIU Bo*, ZHANG Wei*
• 2019, 9(5):  518-526.  DOI: 10.19586/j.2095-2341.2019.0031
• Abstract ( ) PDF (4488KB)( )
Pichia pastoris has become a commonly used eukaryotic expression host due to its multiple advantages including easily genetic manipulation, efficient secretion of recombinant proteins, high-density fermentation and post-translational modification. And P. pastoris expression system based on the methanol-inducible promoter PAOX1 is widely used, however, this system is disadvantageous in production of food enzymes and biopharmaceutical proteins due to flammable and toxic methanol. Therefore, it is of great significance to develop non-toxic inducible expression systems. In previous study, the gene cluster, LRA1~LRA4, was predicted to associate with rhamnose metabolism in P. pastoris, and a rhamnose-inducible promoter PLRA3 was disclosed to drive efficient expression of recombinant proteins in P. pastoris. However, it is necessary to further improve the inducible strength and basal transcriptional profiles of PLRA3. Based on this situation, LRA3 was verified to involve in rhamnose metabolism in P. pastoris via gene disruption and gene complementation, simultaneously the transcription start site of PLRA3 was determined via circularized RNA reverse transcription PCR (CRRT-PCR)  and its cis-acting elements were predicted, which provided some clues for the rational design of PLRA3.
• Effect of Phosphorylation on Deubiquitinase Activity of UCHL3 in Vitro
• DING Shan1, REN Yujing1,2, JIANG Ling1, MEI Ziqing1*
• 2019, 9(5):  527-535.  DOI: 10.19586/j.2095-2341.2019.0029
• Abstract ( ) PDF (9349KB)( )
Ubiquitin C-terminal hydropase-L3 (UCHL3), an important member of the ubiquitin C-terminal hydrolases (UCHs) family, is involved in DNA damage repair and other processes. Previous studies showed that UCHL3 can cleave C-terminal conjugate of ubiquitin with 7-amino-4-methylcoumarin (Ub-AMC) in vitro and the phosphorylation of Ser75 obviously promotes the cleavage activity of UCHL3 towards poly-ubiquitin chains in vivo. But this regulation has not been corroborated by any biochemical data in vitro so far. Based on this situation, wild-type UCHL3 (UCHL3WT) and simulated phosphorylated UCHL3 protein (UCHL3S75E) were prepared by QuikChange site-directed mutagenesis technique and step-wise chromatography methods. The effect of phosphorylation at Ser75 on the poly-ubiquitin chains cleavage activity of UCHL3 in vitro was studied. The results showed that purified UCHL3S75E displayed the enhancing cleavage activity towards Ub-AMC compared to UCHL3WT by 70%, but no physiological level activity to di-ubiquitin (diub) chains, which suggested that the mechanism by which UCHL3 cuts ubiquitin chains might be more complex. Meanwhile, sequence alignment and phylogenetic trees analysis showed that the phosphorylated Ser75 only existed in UCHL3 but not in other UCH family members, indicating that the phosphorylation regulation based on Ser75 probably was unique to UCHL3. In addition, UCHL3 was highly conserved in various eukaryotic organisms, implying that the phosphorylation regulation mechanism of UCHL3 was highly conserved in evolution. The results of this research expanded the understanding of the phosphorylation regulation of UCHL3 and laid a foundation for further understanding of its physiological role.
• The Relationship Between Helicobacter pylori Colonization and Duodenal flora Structure in Duodenal Ulcer
• CHEN Xia1, XIA Chenmei1, DAI Zaiyou2, LI Qianqian1, JIN Lingxiao1, LIN Peili3, ZHANG Zhongchen1, ZHONG Haibing1, YAN Haifan1, WANG Guoping1, DAI Li1, ZHONG Yufen1, WU Zhongbiao4*
• 2019, 9(5):  536-542.  DOI: 10.19586/j.2095-2341.2019.0017
• Abstract ( ) PDF (3895KB)( )
Through the research on the structure of gastric mucosa in patients with Helicobacter pylori (Hp) positive and negative duodenal ulcer, the effects of Hp infection and duodenal ulcer on the structure of duodenal mucosa were investigated. Five cases of duodenal mucosa in patients with Hp-positive and Hp-negative duodenal ulcer were selected. The 16S rRNA gene amplicon sequencing was used to detect the proportion of Hp colonization in duodenal mucosa. Structural analysis was performed. Results showed that, the number of species in the Hp-positive mucosa of patients with duodenal ulcer was significantly higher than that of Hp-negative mucosa (P<0.05), but the difference in microbial diversity was not statistically significant. Except Hp, Prevotella and Neisseria were also the dominatnt flora.  The content of Leptotrichia was significantly different between the two groups (P<0.05). Results indicated that, Hp does not become the only dominant bacteria after infection of duodenal mucosa, but increases the number of species in the duodenal mucosa. And it has little effect on the microbial diversity and has a certain influence on the flora structure. Leptotrichia may be involved in the development of ulcers.