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1.
Rapid detection of potato late blight using a loop-mediated isothermal amplification assay
KONG Liang, WANG Hui-bin, WANG Shuai-shuai, XU Ping-ping, ZHANG Ruo-fang, DONG Suo-meng, ZHENG Xiao-bo
Journal of Integrative Agriculture    2020, 19 (5): 1274-1282.   DOI: 10.1016/S2095-3119(19)62816-9
摘要162)      PDF    收藏
Potato late blight caused by Phytophthora infestans is one of the most destructive plant diseases that threaten global food security.  Early and effective diagnosis of P. infestans is required before disease management decisions are made.  Here, we developed a quick protocol to detect P. infestans based on a loop-mediated isothermal amplification (LAMP) assay.  The P. infestans specific multiple copy DNA sequences (PiSMC), a transposon-like element, provides an ideal target for molecular detection of this pathogen.  We designed highly specific and sensitive primers allowing effective LAMP detection of the pathogen at 64°C in 70 min.  In the validation assay, all 15 P. infestans isolates collected from China, Europe and South America could be positively detected, but results of the other 9 Phytophthora species infecting different plants, fungal and bacterial plant pathogens tested were negative.  The detection limit of this assay is 1 pg P. infestans DNA.  Moreover, the LAMP-PiSMC assay is able to detect P. infestans from infected leaves, tubers and soil.  Taken together, this study reports the development of a specific and sensitive LAMP-PiSMC assay for early diagnosis of potato late blight.
 
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2. Investigation on the co-infections of Toxoplasma gondii with PRRSV, CSFV or PCV-2 in swine in part of China
Wang Shuai, ZHang Meng, LIU Xin-chao, LIn Tao, Yang Han-chun, YUan Shi-shan, ZHao guang-wei, Ia Hassan, Yan Ruo-feng, Song Xiao-kai, XU Li-xin, LI Xiang-rui
Journal of Integrative Agriculture    2015, 14 (9): 1838-1844.   DOI: 10.1016/S2095-3119(15)61044-9
摘要1515)      PDF    收藏
The objective of the present investigation was to estimate the prevalence of Toxoplasma gondii infection and co-infection with porcine reproductive and respiratory syndrome virus (PRRSV), classical swine fever virus (CSFV) and porcine circovirus type 2 (PCV-2) in pigs in China. A total of 372 tissues or serum samples collected from pigs distributed in 9 provinces/ municipalities of China during the period from February 2011 to November 2012 were assayed for T. gondii antigens and antibodies using enzyme linked immunosorbent assay (ELISA) technique, while the PCR was designed for the detection of the PRRSV, CSFV and PCV-2, respectively. The total positive rate of T. gondii, PRSSV, CSFV and PCV-2 was 9.14% (34/372), 50.00% (186/372), 37.10% (138/372) and 3.23% (12/372), respectively. Among the 34 T. gondii positive samples, 26 samples were simultaneously infected with T. gondii and viruses, while the remaining eight samples were infected with T. gondii alone. In addition, the co-infection rate of T. gondii with PRSSV, T. gondii with PRSSV and CSFV, T. gondii with PRSSV and PCV-2, T. gondii with CSFV and PCV-2, T. gondii with PRSSV, CSFV and PCV-2 was 1.61% (6/372), 4.03% (15/372), 0.27% (1/372), 0.27% (1/372) and 0.81% (3/372), respectively. The results of the present survey revealed that PRRSV and CSFV were the common pathogens co-existing with porcine toxoplasmosis in China, and both of them could increase the chances of T. gondii infection in pig. This is the first report of T. gondii co-infections with viruses in pigs. It is very important to understand the interactions of parasite and virus, and can be used as reference data for the control and prevention of co-infections of T. gondii and viruses in pigs.
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3. Type I strain of Toxoplasma gondii from chicken induced different immune responses with that from human, cat and swine in chicken
Zhao Guang-wei, WanG Shuai, WanG Wang, ZhanG Zhen-chao, XIe Qing, ZhanG Meng, I a hassan, Yan Ruo-feng, SonG Xiao-kai, Xu Li-xin, LI Xiang-rui
Journal of Integrative Agriculture    2015, 14 (5): 956-965.   DOI: 10.1016/S2095-3119(14)60861-3
摘要2216)      PDF    收藏
In this study, four strains of Toxoplasma gondii with the same genetic type (Type I) originated from chicken, human, cat and swine were used to compare the immune responses in resistant chicken host to investigate the relationships between the parasite origins and the pathogenicity in certain host. A total of 300, 10-day-old chickens were allocated randomly into five groups which named JS (from chicken), CAT (from cat), CN (from swine), RH (from human) and a negative control group (–Ve) with 60 birds in each group. Tachyzoites of four different T. gondii strains (JS, CAT, CN and RH) were inoculated intraperitoneally with the dose of 1×107 in the four designed groups, respectively. The negative control (–Ve) group was mockly inoculated with phosphate-buffered saline (PBS) alone. Blood and spleen samples were obtained on the day of inoculation (day 0) and at days 4, 11, 25, 39 and 53 post-infection to screen the immunopathological changes. The results demonstrated some different immune characters of T. gondii infected chickens with that of mice or swine previous reported. These differences included up-regulation of major histocompatibility complex class II (MHC II) molecules in the early stage of infection, early peak expressions of interleukin (IL)-12 (IL-12) and -10 (IL-10) and long keep of IL-17. These might partially contribute to the resistance of chicken to T. gondii infection. Comparisons to chickens infected with strains from human, cat and swine, chickens infected with strain from chicken showed significant high levels of CD4+ and CD8+ T cells, interferon gamma (IFN-γ), IL-12 and IL-10. It suggested that the strain from chicken had different ability to stimulate cellular immunity in chicken.
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4. Adsorption of Cu(II) on humic acids derived from different organic materials
LI Cui-lan, JI Fan, WANG Shuai, ZHANG Jin-jing, GAO Qiang, WU Jing-gui, ZHAO Lan-po, WANG Li-chun, ZHENG Li-rong
Journal of Integrative Agriculture    2015, 14 (1): 168-177.   DOI: 10.1016/S2095-3119(13)60682-6
摘要1896)      PDF    收藏
The adsorption of Cu(II) from aqueous solution onto humic acid (HA) which was isolated from cattle manure (CHA), peat (PHA), and leaf litter (LHA) as a function of contact time, pH, ion strength, and initial concentration was studied using the batch method. X-ray absorption spectroscopy (XAS) was used to examine the coordination environment of the Cu(II) adsorbed by HA at a molecular level. Moreover, the chemical compositions of the isolated HA were characterized by elemental analysis and solid-state 13C nuclear magnetic resonance spectroscopy (NMR). The kinetic data showed that the adsorption equilibrium can be achieved within 8 h. The adsorption kinetics followed the pseudo-second-order equation. The adsorption isotherms could be well fitted by the Langmuir model, and the maximum adsorption capacities of Cu(II) on CHA, PHA, and LHA were 229.4, 210.4, and 197.7 mg g–1, respectively. The adsorption of Cu(II) on HA increased with the increase in pH from 2 to 7, and maintained a high level at pH>7. The adsorption of Cu(II) was also strongly influenced by the low ionic strength of 0.01 to 0.2 mol L–1 NaNO3, but was weakly influenced by high ionic strength of 0.4 to 1 mol L–1 NaNO3. The Cu(II) adsorption on HA may be mainly attributed to ion exchange and surface complexation. XAS results revealed that the binding site and oxidation state of Cu adsorbed on HA surface did not change at the initial Cu(II) concentrations of 15 to 40 mg L–1. For all the Cu(II) adsorption samples, each Cu atom was surrounded by 4 O/N atoms at a bond distance of 1.95 Å in the first coordination shell. The presence of the higher Cu coordination shells proved that Cu(II) was adsorbed via an inner-sphere covalent bond onto the HA surface. Among the three HA samples, the adsorption capacity and affinity of CHA for Cu(II) was the greatest, followed by that of PHA and LHA. All the three HA samples exhibited similar types of elemental and functional groups, but different contents of elemental and functional groups. CHA contained larger proportions of methoxyl C, phenolic C and carbonyl C, and smaller proportions of alkyl C and carbohydrate C than PHA and LHA. The structural differences of the three HA samples are responsible for their distinct adsorption capacity and affinity toward Cu(II). These results are important to achieve better understanding of the behavior of Cu(II) in soil and water bodies in the presence of organic materials.
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5. The mRNA Expression Profiles of Five Heat Shock Protein Genes from Frankliniella occidentalis at Different Stages and Their Responses to Temperatures and Insecticides
WANG Hai-hong, Stuart R Reitz, WANG Li-xia, WANG Shuai-yu, LI Xue , LEI Zhong-ren
Journal of Integrative Agriculture    2014, 13 (10): 2196-2210.   DOI: 10.1016/S2095-3119(13)60680-2
摘要1295)      PDF    收藏
The western flower thrips, Frankliniella occidentalis (Pergande) is a highly invasive pest that is able to exploit many crops across a wide range of environmental conditions. Five full-length cDNAs of heat shock protein (HSP) genes (Fo-HSP90, Fo-HSP70, Fo-HSP60, Fo-HSP40 and Fo-HSP28.9) were cloned from F. occidentalis, and their expression profiles were investigated under conditions of thermal stress and insecticide exposure, and at different stages during development, using real-time quantitative PCR. All five gene sequences showed high similarity to homologs in other species, indicating the conserved function of this gene family. HSP60 represents an informative phylogenetic marker at the ordinal taxonomic level within Insecta, but HSP90, which has two homologous copies in Hymenoptera, was not informative. The expression of Fo-HSPs under thermal stress suggests that Fo-HSP90, Fo-HSP70, and Fo-HSP28.9 are inducible by both cold and heat stress, Fo-HSP40 is only heat-inducible, and Fo-HSP60 is thermally insensitive. There were two patterns of cold induction of Fo-HSPs: one is from 0 to 4°C and the other is around -8°C. All five Fo-HSPs genes were induced by exposure to sublethal concentrations of the insecticide avermectin. The expression of the five Fo-HSPs during different developmental stages suggests that they all play a role in development of F. occidentalis.
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