Morphological, Phylogenetic and Biological Characteristics of Ectropisobliqua Single-Nucleocapsid Nucleopolyhedrovirus

Chuan-Xi Zhang,Hai-Jun Xu,Mei-Jun Tang,Qiang Xiao,Jian Hong,Xiu-cui Ma 한국미생물학회 2006 The journal of microbiology Vol.44 No.1

The tea looper caterpillar, Ectropis obliqua, is one of the major pests of tea bushes. E. obliqua single-nucleocapsid nucleopolyhedrovirus (EcobSNPV) has been used as a commercial pesticide for biocontrol of this insect. However only limited genetic analysis for this important virus has been done up to now. EcobSNPV was characterized in this study. Electron microscopy analysis of the occlusion body showed polyhedra of 0.7 to 1.7 μm in diameter containing a single nucleocapsid per envelope of the virion. A 15.5 kb genomic fragment containing EcoRI-L, EcoRI-N and HindIII-F fragments, was sequenced. Analysis of the sequence revealed that the fragment contained eleven potential open reading frames (ORFs): lef-1, egt, 38.7k, rr1, polyhedrin, orf1629, pk-1, hoar and homologues to Spodoptera exigua multicapsid NPV (SeMNPV) ORFs 15, 28, and 29. Gene arrangement and phylogeny analysis suggest that EcobSNPV is closely related to the previously described Group II NPV. Bioassays on lethal concentration (LC50 and LC90) and lethal time (LT50 and LT90) were conducted to test the susceptibility of E. obliqua larvae to the virus.

The utilization and industrialization of insect resources in China

Chuan-Xi Zhang,Xu-Dong TANG,Jia-An CHENG 한국곤충학회 2008 Entomological Research Vol.38 No.-

Insecta is the biggest group of animals on earth. The insects are thought to be one of the biggest biological resources that have not been fully exploited by humans. China is one of the earliest countries to exploit insect resources in the world and has been the top producer for over one thousand years of many insect-related industrial products, such as silk, insect wax and Chinese gallnuts. The exploitation and industrialization of insect resources in China is generally classified into four different levels. The first level is the direct utilization of insect bodies and their secretions, the history of which can be traced back for thousands of years. This level includes the culture and utilization of the silkworm Bombyx mori, the Chinese honeybee Apis cerana cerana, the Chinese white-wax scale Ericerus pela, the Chinese gall aphid Schlechtendalia chinensis, and the lac insects Kerria spp. Additionally, numerous other insects are typically used for Chinese traditional medicines and food, such as Eupolyphaga sinensis, Opisthoplatia orientalis, Aspongopus chinensis, Martianus dermestoides, Polyrhachis vicina, Hepialus spp, Vespa, Hydrillodes repugnalis, and Tenebrio molitor. Pollinators (Megachile rotundata, Osmia cornifrons, O. excavata) and ornamental insects like butterflies, katydids Gampsocleis grafiosa, and fighting crickets Scapsipedes micado are also among the insects included in this level. Accordingly, a related industry is insect-breeding, including sericulture and apiculture, which lays the basis for all insect industrialization. The second level is the utilization of insects as enemies of pests and insect pathogens for biological control. The enemy insects, including the egg parasites tricogramma Trichogramma spp, the seven spotted lady beetle Coccinella septempunctata, the Chinese green lacewing Chrysopa sinica, and Anastatus sp. could be produced in large scale. The insect pathogens that have been extensively used for commercial biocontrol in China include Helicoverpa armigera Nucleopolyhedrovirus (HaSNPV), Ectropis oblique Nucleopolyhedrovirus (EcobNPV), Spodoptera exigua multicapsid nucleopolyhedrovirus (SeMNPV), Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV), Plutella xylostella Granulosis (PxGV), Pieris rapae granulosis (PiraGV), and Bacillus thuringiensis (Bt). Related industries include the biopesticide industry and the enemy insect production industry. The third level of utilization is the extraction and synthesis of insect materials with diverse bioactivities. Some insect pheromones and hormones extracted from insect bodies or chemically synthesized have been used for insect pest control and for regulating the silkworm breeding. Toxins from honeybees and wasps have been used in medicine. Some insect materials from the larvae of honeybees, silkworms, tasar silkworms, and houseflies have been developed into health products. The fourth level is using the insects as bioreactors to produce peptides for medical and veterinary uses. Hundreds of foreign genes have been successfully expressed in the insect cells and larvae. The hGM-CSF expressed in silkworm pupae is commercially available. In this article, we review the culture and utilization of important industrial insects in China.

Characterization of Ha29, a Specific Gene for Helicoverpa armigeraSingle-nucleocapsid Nucleopolyhedrovirus

Chuan-Xi Zhang,Zhong-Jian Guo,Shi-Heng An,Dun Wang,Yan-He Liu,V. Shyam Kumar 한국생화학분자생물학회 2005 BMB Reports Vol.38 No.3

Open reading frame 29 (ha29) is a gene specific for Helicoverpa armigera single-nucleocapsid nucleopolyhedrovirus (HearSNPV). Sequence analyses showed that the transcription factor Tfb2 motif, bromodomain and Half-A-TPR (HAT) repeat were present at aa 66-82, 4-76, 55-90 of the Ha29 protein respectively. The product of Ha29 was detected in HearSNPV-infected HzAM1 cells at 3 h post-infection. Western blot analysis using a polyclonal antibody produced by immunizing a rabbit with purified GST-Ha29 fusion protein indicates that Ha29 is an early gene. The size of Ha29 product in infected HzAM1 cells was about 25 kDa, which was larger than the presumed size of 20.4 kDa. Tunicamycin treatment of HearSNPV-infected HzAM1 cells suggested that the Ha29 protein is N-glycosylated. Fluorescent confocal laser scanning microscope examination, and Western blot analysis of purified budded virus (BVs), occlusion-derived virus (O DVs), cell nuclear and cytoplasmic fraction, showed that the Ha29 protein was localized in the nucleus. Our results suggested that ha29 of HearSNPV encodes a non-structurally functional protein that may be associated with virus gene transcription in Helicoverpa hosts.

Histological Validation of Cardiovascular Magnetic Resonance T1 Mapping for Assessing the Evolution of Myocardial Injury in Myocardial Infarction: An Experimental Study

Zhang Lu,Yang Zhi-gang,Xu Huayan,Yang Meng-xi,Xu Rong,Chen Lin,Sun Ran,Miao Tianyu,Zhao Jichun,Zhou Xiaoyue,Fu Chuan,Guo Yingkun 대한영상의학회 2020 Korean Journal of Radiology Vol.21 No.12

Objective: To determine whether T1 mapping could monitor the dynamic changes of injury in myocardial infarction (MI) and be histologically validated. Materials and Methods: In 22 pigs, MI was induced by ligating the left anterior descending artery and they underwent serial cardiovascular magnetic resonance examinations with modified Look-Locker inversion T1 mapping and extracellular volume (ECV) computation in acute (within 24 hours, n = 22), subacute (7 days, n = 13), and chronic (3 months, n = 7) phases of MI. Masson’s trichrome staining was performed for histological ECV calculation. Myocardial native T1 and ECV were obtained by region of interest measurement in infarcted, peri-infarct, and remote myocardium. Results: Native T1 and ECV in peri-infarct myocardium differed from remote myocardium in acute (1181 ± 62 ms vs. 1113 ± 64 ms, p = 0.002; 24 ± 4% vs. 19 ± 4%, p = 0.031) and subacute phases (1264 ± 41 ms vs. 1171 ± 56 ms, p < 0.001; 27 ± 4% vs. 22 ± 2%, p = 0.009) but not in chronic phase (1157 ± 57 ms vs. 1120 ± 54 ms, p = 0.934; 23 ± 2% vs. 20 ± 1%, p = 0.109). From acute to chronic MI, infarcted native T1 peaked in subacute phase (1275 ± 63 ms vs. 1637 ± 123 ms vs. 1471 ± 98 ms, p < 0.001), while ECV progressively increased with time (35 ± 7% vs. 46 ± 6% vs. 52 ± 4%, p < 0.001). Native T1 correlated well with histological findings (R2 = 0.65 to 0.89, all p < 0.001) so did ECV (R2 = 0.73 to 0.94, all p < 0.001). Conclusion: T1 mapping allows the quantitative assessment of injury in MI and the noninvasive monitoring of tissue injury evolution, which correlates well with histological findings.

Helicoverpa armigera Nucleopolyhedrovirus ORF80 Encodes a Late, Nonstructural Protein

( Dun Wang ),( Chuan Xi Zhang ) 생화학분자생물학회 2007 BMB Reports Vol.40 No.1

BmNPV chitinase gene deletion enhances foreign gene expression in a BmN cell system

Fang WANG,Chuan-Xi ZHANG 한국곤충학회 2009 Entomological Research Vol.39 No.1

Bombyx mori nucleopolyhedrovirus (BmNPV) is extensively being studied as an expression vector for heterologous gene expression in silkworm-derived cells as well as in the host larvae or pupae. BmNPV chitinase is necessary for liquefaction of the virus-infected host insect. The influence of chitinase on the efficiency of foreign gene expression was studied to provide a scientific basis for improving the BmNPV expression system. The BmNPV chitinase gene (chiA) was deleted and the expression level of the polyhedrin promoter controlling the lacZ gene in BmN cells was determined. Sodium dodecylsulfate (SDS)–olyacrylamide gel electrophoresis (PAGE) showed that β-galactosidase accounted for approximately 6.9 and 7.7% of the total protein in BmN cells infected with the chiA deficient Bm lacZ+chiA−,at 3 and 4 days post infection, while the total protein was 3.2 and 4.2% in cells infected with Bm lacZ+. The relative β-galactosidase activities in Bm lacZ+chiA−[기호] -infected cells improved 2.33- and 1.56-fold compared to those of Bm lacZ+-infected cells at 3 and 4 days post infection. The results of the present study suggest that chitinase deletion could improve the lacZ expression level in the BmNPV-BmN cell expression system.

Helicoverpa armigera Nucleopolyhedrovirus ORF80 Encodes a Late, Nonstructural Protein

Wang, Dun,Zhang, Chuan-Xi Korean Society for Biochemistry and Molecular Biol 2007 Journal of biochemistry and molecular biology Vol.40 No.1

The Helicoverpa armigera nucleopolyhedrovirus (HearNPV) ORF80 (ha80) has 765 bp encoding a protein with approximately 254 amino acids and a predicted molecular weight of 30.8 kDa. Homologues of ha80 are found in most baculovirus sequences, including those from lepidopteran NPVs, lepidopteran granuloviruses (GVs), hymenopteran baculoviruses, and one dipteran baculovirus, yet their functions remain unclear. In this study we characterized ha80, and showed that it was transcribed late in infected host cells (HzAM1). The product of ha80 was a 31 kDa protein that was not a structural protein of budded virus (BV) or occlusion-derived virus (ODV) particles. Ha80 was first detected in the cytoplasm of infected HzAM1 cells at 12 h p.i., and was observed in the nucleus at later stages of infection, suggesting that it may be involved in transporting viral proteins into the host cell nucleus or play its roles in the nucleus.

ODV-Associated Proteins of the <i>Pieris rapae</i> Granulovirus

Wang, Xiao-Feng,Zhang, Bao-Qin,Xu, Hai-Jun,Cui, Ying-Jun,Xu, Yi-Peng,Zhang, Min-Juan,Han, Yeon Soo,Lee, Yong Seok,Bao, Yan-Yuan,Zhang, Chuan-Xi American Chemical Society 2011 JOURNAL OF PROTEOME RESEARCH Vol.10 No.6

<P><I>Alphabaculovirus</I> (lepidopteran-specific nucleopolyhedroviruses, NPV) and <I>Betabaculovirus</I> (granuloviruses, GV) are two main genera of the family Baculoviridae. The virion proteomes of <I>Alphabaculovirus</I> have been well studied; however, the <I>Betabaculovirus</I> virion compositions remain unclear. <I>Pieris rapae</I> granulovirus (PrGV) can kill larvae of <I>P. rapae</I>, a worldwide and important pest of mustard family crops. In this study, the occlusion-derived virus (ODV)-associated proteins of PrGV were identified using three mass spectrometry (MS) approaches. The MS analyses demonstrated that 47 proteins were present in PrGV-ODV. Of the 47 PrGV-ODV proteins, 33 have homologues identified previously in other baculovirus ODV/BVs, whereas 14 (P10, Pr21, Pr29, Pr35, Pr42, Pr54, P45/48, Pr83, Pr84, Pr89, Pr92, Pr111, Pr114 and FGF3) were newly identified ODV proteins. Seven of the 14 newly identified ODV proteins are specific to <I>Betabaculovirus</I>, including Pr35, Pr42, Pr54, Pr83, Pr84, Pr111 and Pr114. Furthermore, the data derived from these MS approaches were validated by immunoblotting analysis using antisera prepared from 11 randomly selected recombinant PrGV-ODV proteins (including 5 <I>Betabaculovirus</I>-unique proteins). Comparison analyses revealed the similar and different compositions between <I>Betabaculovirus</I> and <I>Alphabaculovirus</I> virions, which deepen our understanding of the baculovirus virion structure and provide helpful information on <I>Betabaculovirus</I>–host interaction studies.</P><P>We used three mass spectrometry (MS) approaches to identify the occlusion-derived virus (ODV)-associated proteins of the <I>Pieris rapae</I> Granulovirus. A total of 47 proteins were identified; 14 of them were first identified in the ODV, and 7 are specific to Granulovirus.</P><P><B>Graphic Abstract</B> <IMG SRC='http://pubs.acs.org/appl/literatum/publisher/achs/journals/content/jprobs/2011/jprobs.2011.10.issue-6/pr2000804/production/images/medium/pr-2011-000804_0002.gif'></P>

Molecular characterization and inhibition analysis of the acetylcholinesterase gene from the silkworm maggot, Exorista sorbillans

( Guo Jun Lang ),( Ming Yan Zhang ),( Bao Ling Li ),( Lin Lin Yu ),( Xing Meng Lu ),( Chuan Xi Zhang ) 생화학분자생물학회 (구 한국생화학분자생물학회) 2010 BMB Reports Vol.43 No.8

Several organophosphorus (OP) insecticides can selectively kill the silkworm maggot, Exorista sorbillans (Es) (Diptera: Tachinidae), while not obviously affecting the host (Bombyx mori) larvae, but the mechanism is not yet clear. In this study, the cDNA encoding an acetylcholinesterase (AChE) from the field Es was isolated. One point mutation (Gly353Ala) was identified. The Es-353G AChE and Es-353A AChE were expressed in baculovirus- insect cell system, respectively. The inhibition results showed that for eserine and Chlorpyrifos, Es-353A AChE was significantly less sensitive than Es-353G AChE. Meanwhile, comparison of the I(50) values of eserine, dichlorvos, Chlorpyrifos and omethoate of recombinant Es AChEs with its host (Bombyx mori) AChEs indicated that, both Es AChEs are more sensitive than B. mori AChEs. The results give an insight of the mechanism that some OP insecticides can selectively kills Es while without distinct effect on its host, B. mori. [BMB reports 2010; 43(8): 573-578]

Transcriptional analysis of Pieris rapae in response to P. rapae granulovirus

Hai-Jian Huang,Tong-Qiang Zhang,Qiao Lin, Jian-Hui Ye,Chuan-Xi ZHANG,Bao-Qin Zhang 한국응용곤충학회 2018 Journal of Asia-Pacific Entomology Vol.21 No.2

Pieris rapae granulovirus (PrGV) is an important pathogen that has been exploited as a microbial insecticide to control agriculture pests. They can specifically infect cabbage butterfly (Pieris rapae), causing a series of pathological symptoms. In this infected P. rapae at 6 h and 72 h. As a result, a series of host genes were significantly modulated following PrGV infection, including those correlated with exoskeleton, ribosome, heat shock protein (HSP), proteasome, oxidation-reduction and apoptosis. Taken together, our study unveiled the P. rapae response to PrGV at different time point and provided a potential strategy for pest management.