Shuifang Zhu

Chinese Academy of Inspection and Quarantine, Peping, Beijing, China

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Publications (28)91.68 Total impact

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    ABSTRACT: Alien species are brought into countries world wide on a massive scale for agricultural production, ex situ conservation, landscape aesthetics, gardens, and ecosystem restoration. Unfortunately, some of these species have escaped and adversely impacted on regional as well as global biodiversity conservation and agricultural production. To reduce such risks, it is necessary to implement specific and effective measures. Since various government departments and institutions are involved in the management of alien species, it is difficult to prevent native and agroecosystems from being invaded by invited species. We propose the establishment of a supervision and inspection continuum over intentional species introduction, similar to that which exists in some countries over unintentional species introductions. Namely, a justification of the necessity to import, a risk assessment, assurances as to provision of an adequate containment facility assessment, and a damage-limitation protocol should that need to be invoked. These requirements should be satisfied before an alien species is knowingly imported, and the necessary follow-up supervision is important post- importation.
    Biodiversity and Conservation 09/2014; 23(10). · 2.26 Impact Factor
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    ABSTRACT: The aim was to establish an effective screening microarray at genus level for Pospiviroid. We analyzed nucleotide sequences from Pospiviroid viroid and designed 19 probes with genus identification characteristics. The standards of these probes included the characters of (i) a GC content between 40 and 60%, (ii) less than 50% of single nucleotide, (iii) less than 4 continuous mononucleotides, and (iv) less than 6 nucleotides in the inner hairpin. We synthesized microarrays by using these probes on glass slides. The validation results of microarray probes show effective signals from chrysanthemum stunt viroid and tomato planta macho viroid standard samples hybridization. The sensitivity results show that the microarray detected 200 pg/microL of total RNA. The microarray can be used to screen Pospiviroid viroid.
    03/2014; 30(3):514-23.
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    ABSTRACT: Transgenic maize plant expressing high phytase activity has been reported and approved by Chinese government in 2009. Here, we report a highly specific loop-mediated isothermal amplification (LAMP) method to detect the phytase gene in the GMO maize. The LAMP reaction takes less than 20 min and the amplification is visible without gel electrophoresis. The detection sensitivity of the LAMP method is about 30 copies of phytase genomic DNA, which is 33.3 times greater than the conventional PCR method with gel electrophoresis. The quantitative detection results showed that the LAMP method has a good linear correlation between the DNA copy number and the associated Tt values over a large dynamic range of template concentration from 6 × 101 to 6 × 107 copies, with a quantification limit of 60 copies. Therefore, the LAMP method is visual, faster, and more sensitive, and does not need special equipment compared to traditional PCR technique, which is very useful for field tests and fast screening of GMO feeds.
    Food Chemistry 01/2014; 156:184–189. · 3.33 Impact Factor
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    ABSTRACT: A simple and ultrasensitive DNA biosensor was developed utilizing the color and size changes originating from Ligation Chain Reaction (LCR)-based gold nanoparticle assembly as an indicator. Using synthetic DNA of 10-fold serial dilutions as a target, the limit of detection (LOD) was 1.5aM with a range of DNA concentrations from 0.01 to 1000fM by UV-vis absorption and 0.1aM in the range of 0.01-10,000fM by dynamic light scattering (DLS). This developed method is low-cost, rapid, and much lower LOD for the detection of specific short DNA sequences, and the results can be directly determined either by the color or by the DLS. The capability of high-throughput detection with the aid of PCR amplification apparatus can also be realized.
    Biosensors & bioelectronics 08/2013; 52C:8-12. · 5.43 Impact Factor
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    ABSTRACT: An integration event-specific fluorescent liquid bead array was developed for the simultaneous identification of 10 genetically modified (GM) maize, including Bt176, Bt11, MON810, NK603, GA21, MON88017, MON89034, MIR604, T25 and MIR162, as well as one non-GM maize. The system comprised 11 specific oligonucleotide probes labeled with an amino group and coupled to fluorescence-encoded microspheres. To enable fluorescence detection, 11 pairs of primers labeled with biotin at the 5' ends were used. The hybridization signal of biotinylated PCR product to the probe-coupled microspheres was then detected. The limit of detection of this assay was 0.1% for GM maize, which is lower than the current labeling threshold levels enforced in the EU (0.9%). The results of the positive and negative controls were consistent with their expected situation, which showed that the method was highly specific. We detected GM maize in 20 of the 1370 commercial food samples tested, which were labeled as containing maize. The overall sensitivity, specificity, rapidity and high throughput capacity of this liquid chip system suggest that it could provide a significant improvement over current methods, and potentially offer an improved platform for further research into the detection of other GM plants.
    Biosensors & bioelectronics 06/2013; 49C:360-366. · 5.43 Impact Factor
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    ABSTRACT: We report a biosensor based on Surface Plasmon resonance (SPR) for the selective detection of Maize Chlorotic Mottle Virus (MCMV). 11-mercaptoundecanoic acid was applied on a gold surface to form a self-assembled monolayer and a layer of anti-MCMV antibody was crosslinked on the surface for specific recognition of MCMV. The effects of coupling reaction time and antibody concentration on detection sensitivity were studied. The coverage mass change is a function of the concentration of MCMV with a dynamic range from 1 ppb to 1000 ppb. The detection limit is approximately 1 ppb which is about two orders of magnitude higher than that of the exiting ELISA method. The developed SPR sensor showed highly specific recognition for both purified MCMV and crude extracts from real-world samples.
    Analytical Biochemistry 05/2013; · 2.58 Impact Factor
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    ABSTRACT: A major challenge in the agricultural industry is the development of techniques that can screen plant samples for viroid infection. Microarrays are promising in this regard, as their high throughput nature can potentially allow for the detection of a range of viroids in a single test. In this paper we present a microarray that can detect a wide spectrum of all 8 reported viroid genera including 37 known plant viroid species. The array was constructed using an automated probe design protocol which generated a minimal number of probes to detect viroids at the genus level. The designed microarray showed a high specificity and sensitivity when tested with a set of standard virus samples. Finally, the microarray was applied to screen infected field samples, with Hop stunt viroid infection identified as the major disease causing pathogen for an infected citrus sample.
    PLoS ONE 01/2013; 8(5):e64474. · 3.73 Impact Factor
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    Environmental Science & Technology 06/2012; 46(13):7063-4. · 5.26 Impact Factor
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    ABSTRACT: During the summer of 2008 and 2009, massive algal blooms repeatedly broke out in the Yellow Sea of China. These were undoubtedly caused by the accumulations of one or more species in the macroalgal genus Ulva. In previous reports, morphological observation indicated that the species involved in this phenomenon is Ulva prolifera but molecular analyses indicated that the species belongs to an Ulva linza–procera–prolifera (LPP) clade. Correct identification of the bloom species is required to understand and manage the blooms, but the taxonomic status of the bloom species remains unclear. In the current study, the taxonomic status of 22 selected specimens from the Yellow Sea was assessed by using both morphological and molecular (ITS and rbcL sequences) data. In addition, 5S rDNA analyses were performed for those samples clustering in the LPP clade, and phylogenetic tree and ribotype analyses were constructed for determining the possible origin of the bloom. Three free-floating and two attached Ulva species were distinguished and described: Ulva compressa Linnaeus and Ulva pertusa Kjellman were found in free-floating samples; U. linza Linnaeus was found on rocks; and U. prolifera O.F. Müller was found in both habitats. Diversity in free-floating Ulva of the Yellow Sea appears to be greater than previously thought. The dominant free-floating Ulva species, U. prolifera, was not closely related to local populations attached to rocks but was closely related to populations from Japan. KeywordsYellow Sea–Free floating–Rock attached– Ulva –Phylogeny–Morphology
    Journal of Applied Phycology 01/2012; 24(1):97-108. · 2.33 Impact Factor
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    ABSTRACT: We report a biosensor based on a microcantilever that is modified by a specific peptide for highly selective detection of trimethylamine (TMA). The assay is based on binding-induced bending of the peptide functionalized microcantilevers. The sensor is selectively responsive to TMA. The amplitude of microcantilever bending at equilibrium is a function of the concentration of TMA with a dynamic range from 8 ppm to 800 ppm. The detection limit is approximately 8 ppm. There is a good intra-sensor and an acceptable inter-sensor reproducibility as evidenced by the standard deviation of 5% and 15%, respectively.
    Biosensors & bioelectronics 12/2011; 30(1):140-4. · 5.43 Impact Factor
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    ABSTRACT: To clone plasmid from chinaberry witches'-broom phytoplasma and analyse its molecular characterization. Fragments of one plasmid (pCWBFq) in chinaberry witches'-broom phytoplasma-Fuqing strain (CWBFq) were amplified with primer pairs which were designed according to plasmid sequences published on NCBI. Transmembrane domain and subcellular localization predictions of proteins encoded by the plasmid pCWBFq as well as phylogenetic analysis among the plasmid sequences were completed by using bioinformatic softwares. Southern blot analysis was performed to detect the plasmids existed in CWBFq and several other phytoplasmas with the pCWBFq repA probe. One complete plasmid was sequenced from CWBFq. pCWBFq comprised 4446 bp and had a nucleotide content of 73.5% A + T and encoded six proteins. Protein P2, P3, P4 and P5 of pCWBFq contained 3, 2, 1 and 2 tranmembrane domains respectively, and their predicted signal peptide values were 0.989, 0.505, 0.918 and 0.914 respectively. Homologous comparison showed that RepA homology between pCWBFq and other phytoplasmas was between 9.6% -85.6% , however, the homology of different SSB proteins was between 74.0% - 89.4%. Southern blotting with pCWBFq repA probe confirmed the existence of the plasmids in CWBFq. In addition, The hybridizations occurred with paulownia witches'-broom phytoplasma-Nanyang strain (PaWBNy), periwinkle virescence phytoplasma-Hainan stanin (PeVHn), chinaberry witches'-broom phytoplasma-Fuzhou strain (CWBFz) and mulberry dwarf phytoplasma - Puyang strain (MDPy), whereas, no hybridizarions occurred with jujube witches'-broom phytoplasma-Beijing strain (JWBBj), cherry lethal yellows phytoplasma-Xichang strain (CLYXc) and Bischofia polycarpa witches'-broom phytoplasma-Nanchang strain (BiWBNc). The plasmid encoded a replication associated protein (RepA) and a single-stranded DNA binding protein (SSB), which were for the replication of plasmid. Four putative proteins encoded by the plasmid were predicted to contain one or more hydrophobic transmembrane domains, respectively, and presumably to be localized to the membrane. The alignment and homology analysis as well as phylogenetic analysis to the DNA and encoded protein amino acid sequences of the whole plasmids and single ORFs on the known phytoplasmal plasmids showed that the different homologous sequences have distinct variation, among which the repA gene with the largest diversity appeared in all the known plasmids while ssb with less variation were only found in 16SrI plasmids. CWBFq, PaWBNy, PeVHn, CWBFz and MDPy possessed distinct plasmids in terms of number and size, whereas there was no plasmid detected in JWBBj, CLYXc and BiWBNc, perhaps as a result of low homology among repA genes in plasmids of JWBBj, CLYXc and BiWBNc.
    ACTA MICROBIOLOGICA SINICA 09/2011; 51(9):1158-67.
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    ABSTRACT: Traditional real-time polymerase chain reaction (PCR) requires a purified DNA sample for PCR amplification and detection. This requires PCR tests be conducted in clean laboratories, and limits its applications for field tests. This work developed a method that can carry out DNA purification, amplification and detection in a single PCR tube. The polypropylene PCR tube was first treated with chromic acid and peptide nucleic acids (PNA) as DNA-capturer were immobilized on the internal surface of the tube. Cauliflower mosaic virus 35S (CaMV-35S) promoter in the crude extract was hybridized with the PNA on the tube surface, and the inhibitors, interfering agents and irrelevant DNA in the crude extract were effectively removed by rinsing with buffer solutions. The tube that has captured the target DNA can be used for the following real-time PCR (RT-PCR). By using this approach, the detection of less than 2500 copies of 35S plasmids in a complex sample could be completed within 3 hours. Chocolate samples were tested for real sample analysis, and 35S plasmids in genetically modified chocolate samples have been successfully identified with this method in situ. The novel One-PCR-tube method is competitive for commercial kits with the same time and simpler operation procedure. This method may be widely used for identifying food that contains modified DNA and specific pathogens in the field.
    The Analyst 08/2011; 136(20):4254-9. · 4.23 Impact Factor
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    ABSTRACT: Acidovorax avenae subsp. citrulli (AAC) is one of the most harmful diseases in cucurbit production. A rapid and sensitive DNA strip sensor was constructed based on gold nanoparticle-labeled oligonucleotide probes for the detection of AAC. Both the qualitative and semi-quantitative detections of target DNA were successfully achieved using the developed DNA strip sensor. The qualitative limit of detection (LOD) of the strip sensor was determined as 4 nM. The LOD for the semi-quantitative detection was calculated to be 0.48 nM in the range of 0-10 nM. The genomic DNA was detected directly using the DNA strip sensor without any further treatment. This DNA strip sensor is a potentially useful tool for rapid on-site DNA screening.
    Biosensors & bioelectronics 06/2011; 26(10):4241-4. · 5.43 Impact Factor
  • Yi Yang, Wenjun Zhao, Zhihong Li, Shuifang Zhu
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    ABSTRACT: Amaranth (Amaranthus retroflexus L.) is a common weed that grows vigorously in orchards, roadside verges, fields, woods and scrubland in China. In 2009, phytoplasma disease surveys were made in orchards in Beijing, China, and stem/leaf tissues were collected from asymptomatic amaranths. Direct PCR using universal phytoplasma primers P1/P7 detected 16S rRNA gene sequences in every DNA sample extracted from the symptomless amaranths. Sequence alignment and phylogenetic analyses of the 16S rRNA gene determined that the amaranth phytoplasma strain was related to ‘Candidatus Phytoplasma ziziphi’. Furthermore, virtual RFLP pattern analysis showed that the amaranth phytoplasma belonged to the 16SrV-B subgroup. This is the first report of symptomless plants containing a ‘Candidatus Phytoplasma ziziphi’-related strain.
    Journal of Phytopathology 05/2011; 159(9):635 - 637. · 1.00 Impact Factor
  • Yi Yang, Wenjun Zhao, Zhihong Li, Shuifang Zhu
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    ABSTRACT: Amaranth (Amaranthus retroflexus L.) is a common weed that grows vigorously in orchards, roadside verges, fields, woods and scrubland in China. In 2009, phytoplasma disease surveys were made in orchards in Beijing, China, and stem/leaf tissues were collected from asymptomatic amaranths. Direct PCR using universal phytoplasma primers P1/P7 detected 16S rRNA gene sequences in every DNA sample extracted from the symptomless amaranths. Sequence alignment and phylogenetic analyses of the 16S rRNA gene determined that the amaranth phytoplasma strain was related to ‘Candidatus Phytoplasma ziziphi’. Furthermore, virtual RFLP pattern analysis showed that the amaranth phytoplasma belonged to the 16SrV‐B subgroup. This is the first report of symptomless plants containing a ‘Candidatus Phytoplasma ziziphi’‐related strain.
    Journal of Phytopathology 01/2011; 159(9). · 1.00 Impact Factor
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    ABSTRACT: The fluorescence-encoded microspheres were prepared by the simple and controllable microfluidic device for the first time. The polycaprolactone (PCL) microspheres were encoded with different quantum dots (QDs) at various ratios, and the fluorescence was decoded successfully. The designed fluorescence encoding method was easily manipulated, and generated the uniform microspheres with different size. This research demonstrated a very facile and promising technique for fluorescence encoding over traditional methods.
    Journal of Colloid and Interface Science 12/2010; 352(2):337-42. · 3.17 Impact Factor
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    ABSTRACT: Maize chlorotic mottle virus (MCMV) causes corn lethal necrosis disease, and can be transmitted through infected maize seeds. It remains a challenge to detect this virus in the seeds to prevent its introduction and infection. For this purpose, a real-time TaqMan RT-PCR procedure for efficient detection of MCMV was developed. The sensitivity of the method was 4 fg of total RNA or 25 copies of RNA transcripts, which was approximately ten-fold higher than conventional RT-PCR gel electrophoresis method. The successful detection of MCMV in maize seeds suggested the feasibility of this procedure for routine testing.
    Journal of virological methods 11/2010; 171(1):292-4. · 2.13 Impact Factor
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    ABSTRACT: An ultrasensitive and rapid electrochemical platform for the specific detection of ochratoxin A (OTA) was developed. In this method, three single-stranded DNA molecules, including the aptamer, were immobilized on the surface of an electrode. Binding of the OTA target analyte to the aptamer changed the redox current of methylene blue (MB), which was used as the electrochemical probe, in a manner that was dependent on OTA concentration. With signal enhancement from gold nanoparticle-functionalized DNA, the sensitivity of this method for OTA was as low as 30 pg/mL, and the effective sensing range was from 0.1 to 20 ng/mL. To investigate the sensing process, the conformational switch of the aptamer was studied by circular dichroism (CD), which confirmed the recognition of the aptamer by the target OTA. Given its sensitivity and rapid detection, we believe this approach has the potential to be a main technology for the detection of toxins in the field of food safety, and in other areas.
    Biosensors & bioelectronics 10/2010; 26(2):710-6. · 5.43 Impact Factor
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    ABSTRACT: Jujube witches' -broom (JWB) is an important plant disease caused by phytoplasma. The major objective of our research was to classify JWB in Beijing and Hebei districts and to provide reference for classification in subgroup level. By use of PCR, the elongation factor Tu (tuf gene) and ribosomal protein (rp) gene of phytoplasma associated with JWB in Beijing and Hebei districts were amplified separately with universal primer pairs fTufu/rTufu and rp(v)F1A/rp(v) R1A. Partial tuf gene and rp gene were sequenced and similarity analysed with other phytoplasmas. We obtained partial tuf gene sequence (824bp) and complete rp gene (1196bp) from the diseased sample. In tuf gene, JWB in Beijing shared most similarity (92.84%) with Flavescence dorée (FD) phytoplasma (Candidatus Phytoplasma vitis), however, shared a low similarity (57.29%) with JWB in Shaanxi district which had been already reported. The similarity analysis for sequences of rp gene showed a high identity (> 96%) with members of the 16SrV group phytoplasmas. It shared most identity (99.83%) with JWB strain Taishan and Hemp fiber witches' -broom phytoplasma (HFWB) of the 16SrV group. The JWB strains in Beijing and Hebei are members of 16Sr V; JWB in Beijing and Hebei share high similarity, and show a diversity with JWB in Shaanxi.
    ACTA MICROBIOLOGICA SINICA 10/2010; 50(10):1313-9.
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    ABSTRACT: A major challenge facing agriculture at present is the development of techniques that can screen field samples and other plant materials simultaneously for the presence of many viruses. Microarray techniques show promise in this regard, as their high throughput nature can potentially detect a range of viruses using a single test. In this paper we present an array that can detect a wide spectrum of 169 plant virus species from 13 different genera. The array was constructed using an automated probe design protocol which generated a minimal number of probes to detect viruses at the genus level. The designed arrays showed a high specificity and sensitivity when tested with a set of standard virus samples. Field samples collected from a severe disease outbreak of Panax notoginseng farms in Yunnan, China, in 2001 were screened, where a potyvirus infection was identified associated with the disease.
    Journal of virological methods 03/2010; 167(1):53-60. · 2.13 Impact Factor

Publication Stats

129 Citations
91.68 Total Impact Points

Institutions

  • 2008–2014
    • Chinese Academy of Inspection and Quarantine
      Peping, Beijing, China
  • 2008–2013
    • Jiangnan University
      • School of Food Science and Technology
      Wuxi, Jiangsu Sheng, China
  • 2011
    • China Agricultural University
      • College of Agronomy and Biotechnology
      Beijing, Beijing Shi, China