2017 Vol. 32, No. 8
Display Method:
2017, 32(8): 813-817.
doi: 10.19303/j.issn.1008-0384.2017.08.001
Abstract:
A strain of duck hepatitis A virus 1 subtype a (DHAV-1a), coded as FJ1605, was isolated from the mule ducklings with pancreatitis. It was identified by RT-PCR analysis and a neutralization test in duck embryos that showed the strain specifically being neutralized by the anti-DHAV-1a hyperimmune serum. Molecular characteristics of VP1 gene of the virus was analyzed to indicate the nucleotide to be 714 bp in length with a homology with those of other DHAV-1a isolates ranging from 98.1% to 99.7% and sharing the greatest similarity to strain FJ1220.On the other hand, the homology between FJ1605 and DHAV-2 was merely 65.7%; and, that between FJ1605 and DHAV-3, approximately 68.3%. Phylogenetic analysis on the VP1 gene also showed FJ1605 to be in the viral lineage of DHAV-1a. Furthermore, an animal infection test clearly displayed the same clinical symptoms as those shown inclinical cases. Thus, it was concluded that the pancreatitis in the mule ducklings was caused by DHAV-1a.
A strain of duck hepatitis A virus 1 subtype a (DHAV-1a), coded as FJ1605, was isolated from the mule ducklings with pancreatitis. It was identified by RT-PCR analysis and a neutralization test in duck embryos that showed the strain specifically being neutralized by the anti-DHAV-1a hyperimmune serum. Molecular characteristics of VP1 gene of the virus was analyzed to indicate the nucleotide to be 714 bp in length with a homology with those of other DHAV-1a isolates ranging from 98.1% to 99.7% and sharing the greatest similarity to strain FJ1220.On the other hand, the homology between FJ1605 and DHAV-2 was merely 65.7%; and, that between FJ1605 and DHAV-3, approximately 68.3%. Phylogenetic analysis on the VP1 gene also showed FJ1605 to be in the viral lineage of DHAV-1a. Furthermore, an animal infection test clearly displayed the same clinical symptoms as those shown inclinical cases. Thus, it was concluded that the pancreatitis in the mule ducklings was caused by DHAV-1a.
2017, 32(8): 818-822.
doi: 10.19303/j.issn.1008-0384.2017.08.002
Abstract:
This study aimed to obtain the monoclonal antibodies against porcine epidemic diarrhea virus(PEDV) for antibody characterization as well asdisease control.The antigen of PEDV was purifiedby means of differential centrifugation and sucrose gradientcentrifugation. Spleen cells of the Balb/c mice immunized with viral antigen were fused with SP2/0 myeloma cells.Hybridoma cell lines secreting the monoclonal antibodies were isolated by using the indirect ELISA and sub-cloning approach to provide the source for the target antibody.Subsequently, specificities of the antibody were determined by means of IFA, ELISA and the Western blotting. The results obtained the following. (a) Two isolated hybridoma cell lines that could steadily secretethe specific McAbs against PEDV were designated as E1 and H6. (b) The isotyping analysis showed that E1 belonged to IgG2a; and H6, IgM sub-type. The McAbs from them could both bind to the antigen of PED in ELISA, IFA and the western blot analyses. (c) ELISA titers of the supernatants of E1 and H6 were 26 and 24 respectively; while those of the ascites fluids of E1 and H6, 105 and 104 respectively. (d) The McAbs from E1 and H6 showed no cross-reactivity with Vero cells, CSFV, TGEV or PRRSV.(e) The Western blotting confirmed that E1 McAbs recognized M protein; and, H6 McAbs, N protein. It appeared that two monoclonal antibodies with a high specificity against PEDV were successfully prepared, and further studies on PEDV immunodiagnosis, epitope discernment and the proteins became possible.
This study aimed to obtain the monoclonal antibodies against porcine epidemic diarrhea virus(PEDV) for antibody characterization as well asdisease control.The antigen of PEDV was purifiedby means of differential centrifugation and sucrose gradientcentrifugation. Spleen cells of the Balb/c mice immunized with viral antigen were fused with SP2/0 myeloma cells.Hybridoma cell lines secreting the monoclonal antibodies were isolated by using the indirect ELISA and sub-cloning approach to provide the source for the target antibody.Subsequently, specificities of the antibody were determined by means of IFA, ELISA and the Western blotting. The results obtained the following. (a) Two isolated hybridoma cell lines that could steadily secretethe specific McAbs against PEDV were designated as E1 and H6. (b) The isotyping analysis showed that E1 belonged to IgG2a; and H6, IgM sub-type. The McAbs from them could both bind to the antigen of PED in ELISA, IFA and the western blot analyses. (c) ELISA titers of the supernatants of E1 and H6 were 26 and 24 respectively; while those of the ascites fluids of E1 and H6, 105 and 104 respectively. (d) The McAbs from E1 and H6 showed no cross-reactivity with Vero cells, CSFV, TGEV or PRRSV.(e) The Western blotting confirmed that E1 McAbs recognized M protein; and, H6 McAbs, N protein. It appeared that two monoclonal antibodies with a high specificity against PEDV were successfully prepared, and further studies on PEDV immunodiagnosis, epitope discernment and the proteins became possible.
2017, 32(8): 823-827.
doi: 10.19303/j.issn.1008-0384.2017.08.003
Abstract:
A double antibody sandwich ELISA (DAS-ELISA) was developed using the high specificity, mouse-derived monoclonal antibody (Mab) as the capture antibody and the rabbit-derived polyclonal antibody against porcine epidemic diarrhea virus (PEDV) as the detecting antibody. The optimal reaction conditions for DAS-ELISA was determined to include a coating concentration of 4.40 g·mL-1 for PEDV MAb E1 with 1 h incubation at 37℃, the use of 5% BSA solution for blocking for 1 h, an application of 5.91 μg·mL-1 in concentration of rabbit polyclonal antibodies against PEDV, a 2 000×dilution of HRP, and the positive OD equal or greater than 0.381 at 450 nm wave length on the spectrophotometer measurement. The developed method showed no cross-reaction between porcine rotavirus and transmissible gastroenteritis virus. The detection sensitivity of the method was 30 g·mL-1(5×103.12); and, the coefficient variation of repetition, less than 10%. Furthermore, a total of 42 clinical samples were positively detected by the method in conjunction with RT-PCR at a rate of 92.30%. Consequently, it was concluded that the newly developed DAS-ELISA methodology was highly specific, sensitive, rapid, and hence, applicable for PEDV detection.
A double antibody sandwich ELISA (DAS-ELISA) was developed using the high specificity, mouse-derived monoclonal antibody (Mab) as the capture antibody and the rabbit-derived polyclonal antibody against porcine epidemic diarrhea virus (PEDV) as the detecting antibody. The optimal reaction conditions for DAS-ELISA was determined to include a coating concentration of 4.40 g·mL-1 for PEDV MAb E1 with 1 h incubation at 37℃, the use of 5% BSA solution for blocking for 1 h, an application of 5.91 μg·mL-1 in concentration of rabbit polyclonal antibodies against PEDV, a 2 000×dilution of HRP, and the positive OD equal or greater than 0.381 at 450 nm wave length on the spectrophotometer measurement. The developed method showed no cross-reaction between porcine rotavirus and transmissible gastroenteritis virus. The detection sensitivity of the method was 30 g·mL-1(5×103.12); and, the coefficient variation of repetition, less than 10%. Furthermore, a total of 42 clinical samples were positively detected by the method in conjunction with RT-PCR at a rate of 92.30%. Consequently, it was concluded that the newly developed DAS-ELISA methodology was highly specific, sensitive, rapid, and hence, applicable for PEDV detection.
2017, 32(8): 828-832.
doi: 10.19303/j.issn.1008-0384.2017.08.004
Abstract:
The study was aimed at developing a quantitative real-time fluorescence reverse transcriptase PCR(qRT-PCR) assay according to the conserved gene sequence of BVDV 5'-UTR for detection of bovine viral diarrhea virus(BVDV) obtained from 359 slaughtered swine serum samples of slaughterhouses in Fuzhou Prefecture. The results showed that the BVDV positive rate of slaughtered swines was 14.5%(52 positive samples out of 359 ones). Sequence analysis on 5'-UTR fragments of 13 BVDV positive samples showed that 5'-UTR fragments of all tested samples were closely related to VEDEVAC evolution lineages, all of which were genotype I. It was indicated that the specific qRT-PCR method established based on BVDV was suitable for detection of BVDV in swines. And the BVDV infection was common in slaughtered swines of Fuzhou Prefecture.
The study was aimed at developing a quantitative real-time fluorescence reverse transcriptase PCR(qRT-PCR) assay according to the conserved gene sequence of BVDV 5'-UTR for detection of bovine viral diarrhea virus(BVDV) obtained from 359 slaughtered swine serum samples of slaughterhouses in Fuzhou Prefecture. The results showed that the BVDV positive rate of slaughtered swines was 14.5%(52 positive samples out of 359 ones). Sequence analysis on 5'-UTR fragments of 13 BVDV positive samples showed that 5'-UTR fragments of all tested samples were closely related to VEDEVAC evolution lineages, all of which were genotype I. It was indicated that the specific qRT-PCR method established based on BVDV was suitable for detection of BVDV in swines. And the BVDV infection was common in slaughtered swines of Fuzhou Prefecture.
2017, 32(8): 833-836.
doi: 10.19303/j.issn.1008-0384.2017.08.005
Abstract:
Tissue samples of liver and spleen from the Muscovy ducks that were suspected to have died from the duck plague were collected for this study.Viruses were isolated from the Muscovy duck embryo fibroblasts (MDEF) for a preliminary identification using hemagglutination assay(HA), immunofluorescence analysis(IFA), qPCR, PCR product sequencing, and animal infection test. Four virus strains, DPVfj1, DPVfj2, DPVfj3 and DPVfj4, were thus identified for further investigation. Subsequently, it was found that these strains (a) would not cause red blood cell agglutination on pigeon erythrocytes; (b) were tested negative on IFA with goose parvovirus, Muscovy duck parvovirus, duck reovirus, duck paramyxovirus, and Tembusu virus; (c) showed positive on test with fluorescence RT-PCR kit; (d) had a positive result on their PCR-amplified pair of primers from the specific fragment of gJ protein gene, JQ673560, and a homologygreater than 99% on the sequence of the fragment with that of the duck plague virus(DPV) gJ protein gene; (e)infected 30-day-old ducks by injection and 5-day-old ducklings by co-habitation showing exactly the same clinical symptoms and postmortem signsas in the natural cases; and, (f)were recovered from the diseased birds. The results seemed to verify the fact that DPV caused the duck plaque in natural environment, and that the isolated strains possessed the virulent nature in question.
Tissue samples of liver and spleen from the Muscovy ducks that were suspected to have died from the duck plague were collected for this study.Viruses were isolated from the Muscovy duck embryo fibroblasts (MDEF) for a preliminary identification using hemagglutination assay(HA), immunofluorescence analysis(IFA), qPCR, PCR product sequencing, and animal infection test. Four virus strains, DPVfj1, DPVfj2, DPVfj3 and DPVfj4, were thus identified for further investigation. Subsequently, it was found that these strains (a) would not cause red blood cell agglutination on pigeon erythrocytes; (b) were tested negative on IFA with goose parvovirus, Muscovy duck parvovirus, duck reovirus, duck paramyxovirus, and Tembusu virus; (c) showed positive on test with fluorescence RT-PCR kit; (d) had a positive result on their PCR-amplified pair of primers from the specific fragment of gJ protein gene, JQ673560, and a homologygreater than 99% on the sequence of the fragment with that of the duck plague virus(DPV) gJ protein gene; (e)infected 30-day-old ducks by injection and 5-day-old ducklings by co-habitation showing exactly the same clinical symptoms and postmortem signsas in the natural cases; and, (f)were recovered from the diseased birds. The results seemed to verify the fact that DPV caused the duck plaque in natural environment, and that the isolated strains possessed the virulent nature in question.
2017, 32(8): 837-841.
doi: 10.19303/j.issn.1008-0384.2017.08.006
Abstract:
The pathogen that caused a disease characterized by thin nasal discharge on goats in Fujian during 2014-2017 was investigated.An epidemiological survey of the disease was conducted with tumor tissue biopsy, specific PCR diagnosis, and nucleic acid sequence analysis. It was found that the disease had basically the sam eepidemiology, clinical symptoms, pathology, and histopathology as the similar cases reported domestically and abroad. RT-PCR identified the pathogen to be ENTV-2. Among the various isolated viruses, 7 were in the same phylogenetic tree branch as ENTV-2 CHN2, and one in the branch as ENTV-SC and ENTV-Shaanxi, which were all isolated in China.The results confirmed ENTV-2 to be the culprit of the disease in goats found in Fujian Province. Hopefully, with the information, a prevention measure and/or cure for the disease could be developed in the near future.
The pathogen that caused a disease characterized by thin nasal discharge on goats in Fujian during 2014-2017 was investigated.An epidemiological survey of the disease was conducted with tumor tissue biopsy, specific PCR diagnosis, and nucleic acid sequence analysis. It was found that the disease had basically the sam eepidemiology, clinical symptoms, pathology, and histopathology as the similar cases reported domestically and abroad. RT-PCR identified the pathogen to be ENTV-2. Among the various isolated viruses, 7 were in the same phylogenetic tree branch as ENTV-2 CHN2, and one in the branch as ENTV-SC and ENTV-Shaanxi, which were all isolated in China.The results confirmed ENTV-2 to be the culprit of the disease in goats found in Fujian Province. Hopefully, with the information, a prevention measure and/or cure for the disease could be developed in the near future.
2017, 32(8): 842-848.
doi: 10.19303/j.issn.1008-0384.2017.08.007
Abstract:
Effects of variety and cultivation conditions on production and yield factors of ratoon rice were studied. Various varieties of ratoon rice, stubble height for ratooning, and fertilization to promote budding were implemented in a L9 (34) orthogonal experimentation to optimize the annual crop harvestof the first and second season ratoon rice. It was found that 10 cm-tall stubbles of Liangyou 616 fertilized with N 140 kg·hm-2 would render the highest yield on both crops. The annual production of ratoon rice was affected in the order of:variety > stubble height > bud fertilization, with rice variety playing a prominent role.Grain yield of the first-season crop was affected byvariety, bud fertilizationand stubble heightin that order, with rice variety being the predominant factor as well.Duringthe second season, thegrain and paniclecountson the ratoon rice plants were affected by variety > stubble height > bud fertilization; while, the 1 000-grain-weight and seed setting rate, variety > bud fertilization > stubble height.Within the scope of this experiment, the aforementioned combination of variety choice, stubble height and fertilization wasrecommended for high outputof the ratoon rice cultivation.
Effects of variety and cultivation conditions on production and yield factors of ratoon rice were studied. Various varieties of ratoon rice, stubble height for ratooning, and fertilization to promote budding were implemented in a L9 (34) orthogonal experimentation to optimize the annual crop harvestof the first and second season ratoon rice. It was found that 10 cm-tall stubbles of Liangyou 616 fertilized with N 140 kg·hm-2 would render the highest yield on both crops. The annual production of ratoon rice was affected in the order of:variety > stubble height > bud fertilization, with rice variety playing a prominent role.Grain yield of the first-season crop was affected byvariety, bud fertilizationand stubble heightin that order, with rice variety being the predominant factor as well.Duringthe second season, thegrain and paniclecountson the ratoon rice plants were affected by variety > stubble height > bud fertilization; while, the 1 000-grain-weight and seed setting rate, variety > bud fertilization > stubble height.Within the scope of this experiment, the aforementioned combination of variety choice, stubble height and fertilization wasrecommended for high outputof the ratoon rice cultivation.
2017, 32(8): 849-853.
doi: 10.19303/j.issn.1008-0384.2017.08.008
Abstract:
A ratoon-rice, Tianyouhuazhan, was used to study the effects of N application rates on the photosynthetic products and grain yield of the first-season and ratoon crops. Five rates of N fertilization, i.e., N0 at 0 kg·hm-2 (control), N1 at 69.00 kg·hm-2, N2 at 138.00 kg·hm-2, N3 at 207.00 kg·hm-2, and N4 at 276.00 kg·hm-2, were applied on the first crop. The fields were followed with a 160.50 kg·hm-2 application rate for the ratooning. The results showed that higher N fertilizations increased the tiller number, dry matter accumulation, and transporting of storage substances to the grains, but large amount nitrogen use decreased the N agricultural use efficiency (NAUE) on the first crop. The grain yields increased by 12.12% to 23.35% over that of N0 due to the fertilizations, with the greatest production achieved by N3. The panicle count and spikelets per panicle increased with the rate increases before reaching N4 level. For the ratoon crop, the increases on grain yield ranged from 1.61% to 5.54% as compared with N0. N4 also caused a yield reduction of 4.73% over control. The panicle count of the ratoon crop decreased with rising N application, while the spikelets per panicle increased. It appeared that a N fertilization of 174.12 kg·hm-2 for the first crop followed by 160.50 kg·hm-2 for the ratoon crop could maximize the combined grain production.
A ratoon-rice, Tianyouhuazhan, was used to study the effects of N application rates on the photosynthetic products and grain yield of the first-season and ratoon crops. Five rates of N fertilization, i.e., N0 at 0 kg·hm-2 (control), N1 at 69.00 kg·hm-2, N2 at 138.00 kg·hm-2, N3 at 207.00 kg·hm-2, and N4 at 276.00 kg·hm-2, were applied on the first crop. The fields were followed with a 160.50 kg·hm-2 application rate for the ratooning. The results showed that higher N fertilizations increased the tiller number, dry matter accumulation, and transporting of storage substances to the grains, but large amount nitrogen use decreased the N agricultural use efficiency (NAUE) on the first crop. The grain yields increased by 12.12% to 23.35% over that of N0 due to the fertilizations, with the greatest production achieved by N3. The panicle count and spikelets per panicle increased with the rate increases before reaching N4 level. For the ratoon crop, the increases on grain yield ranged from 1.61% to 5.54% as compared with N0. N4 also caused a yield reduction of 4.73% over control. The panicle count of the ratoon crop decreased with rising N application, while the spikelets per panicle increased. It appeared that a N fertilization of 174.12 kg·hm-2 for the first crop followed by 160.50 kg·hm-2 for the ratoon crop could maximize the combined grain production.
2017, 32(8): 854-858.
doi: 10.19303/j.issn.1008-0384.2017.08.009
Abstract:
Factors affecting the properties of polyphenol oxidase (PPO) and peroxidase (POD) in luffa (Luffa cylindrica) were studied for information relating to enzymatic browning of the vegetable. The temperature, time duration, pH, and concentrations of the substrates, i.e., catechol for PPO activity and guaiacol for POD activity determinations, applied for the reaction as well as the wavelength used for the spectrophotometric color measurements were analyzed. The results indicated that (1) the optimum reaction system for PPO activity determination required the applications of wavelength at 408 nm for the spectrophotometry, a temperature at 35, a pH of 6.0, and an enzyme dosage of 0.15 mL with 0.056 moL·L-1 catechol; and, (2) the optimum reaction system for POD, wavelength at 410 nm, a temperature at 40, a pH of 5.5, an enzyme dosage of 0.1 mL with 0.067 moL·L-1 guaiacol.
Factors affecting the properties of polyphenol oxidase (PPO) and peroxidase (POD) in luffa (Luffa cylindrica) were studied for information relating to enzymatic browning of the vegetable. The temperature, time duration, pH, and concentrations of the substrates, i.e., catechol for PPO activity and guaiacol for POD activity determinations, applied for the reaction as well as the wavelength used for the spectrophotometric color measurements were analyzed. The results indicated that (1) the optimum reaction system for PPO activity determination required the applications of wavelength at 408 nm for the spectrophotometry, a temperature at 35, a pH of 6.0, and an enzyme dosage of 0.15 mL with 0.056 moL·L-1 catechol; and, (2) the optimum reaction system for POD, wavelength at 410 nm, a temperature at 40, a pH of 5.5, an enzyme dosage of 0.1 mL with 0.067 moL·L-1 guaiacol.
2017, 32(8): 859-863.
doi: 10.19303/j.issn.1008-0384.2017.08.010
Abstract:
To select desirable parents for cross-pollination, effect of severalvarieties of Fuguihong pitaya (Hylocereus undulatus Britt) on the properties of the fruits was studied. The appearance, eating quality, fruiting, cracking and development time of the fruits, as well as characteristics of the seeds produced from the flowerscross-pollinated between Fuguihong and 6 varieties of pitaya were compared.Significant improvements on fruiting percentage, single fruit weight, seed count and weight per thousand grains, as well asreductions onincidents of fruit cracking were found because of the cross-pollination. Among the pollen-donor plants, Mihong, Jinduyihao and Dahong also provided the resulting fruits with large edible portion, high soluble solids, and attractive flesh color and flavor; while, Mihong, Jinduyihao, Dahong and Xixianghong, increased totalsugars, vitamin C, and total amino acids, butreduced total acid.Therefore, it appeared that using Mihong, Jinduyihao and/or Dahong pitaya to pollinate the flowers on Fuguihong could enhancethe fruit qualities.
To select desirable parents for cross-pollination, effect of severalvarieties of Fuguihong pitaya (Hylocereus undulatus Britt) on the properties of the fruits was studied. The appearance, eating quality, fruiting, cracking and development time of the fruits, as well as characteristics of the seeds produced from the flowerscross-pollinated between Fuguihong and 6 varieties of pitaya were compared.Significant improvements on fruiting percentage, single fruit weight, seed count and weight per thousand grains, as well asreductions onincidents of fruit cracking were found because of the cross-pollination. Among the pollen-donor plants, Mihong, Jinduyihao and Dahong also provided the resulting fruits with large edible portion, high soluble solids, and attractive flesh color and flavor; while, Mihong, Jinduyihao, Dahong and Xixianghong, increased totalsugars, vitamin C, and total amino acids, butreduced total acid.Therefore, it appeared that using Mihong, Jinduyihao and/or Dahong pitaya to pollinate the flowers on Fuguihong could enhancethe fruit qualities.
2017, 32(8): 864-869.
doi: 10.19303/j.issn.1008-0384.2017.08.011
Abstract:
The natural habitat, phenophases, and morphological characteristics of wild Pholidota chinensis Lindl were studied. The on-site environmental factors, such as temperature, light intensity, and humidity at the area where the plants were located, and the morphological characteristics, such as leaf length and width, pseudobulb length and counts per plant, and rhizome length and girth, of the plants were recorded. The results showed that P. chinensis grew in shady locations along a stream; flowered in April through May; developed fruits in October which ripened in following March; and, underwent no apparent dormancy period in a year.
The natural habitat, phenophases, and morphological characteristics of wild Pholidota chinensis Lindl were studied. The on-site environmental factors, such as temperature, light intensity, and humidity at the area where the plants were located, and the morphological characteristics, such as leaf length and width, pseudobulb length and counts per plant, and rhizome length and girth, of the plants were recorded. The results showed that P. chinensis grew in shady locations along a stream; flowered in April through May; developed fruits in October which ripened in following March; and, underwent no apparent dormancy period in a year.
2017, 32(8): 870-873.
doi: 10.19303/j.issn.1008-0384.2017.08.012
Abstract:
The authenticity of hybrids created by reciprocal crossing between Dendrobium officinale and D. huoshanense was not always apparent. To resolve the confusion, this study applied SSR markers to differentiate them among 63 F1 generation of the hybrids. Three out of 21 pairs of the SSR primers were found to have polymorphic bands, i.e., 14.3% of all, in the parental lines. By using these 3 pairs of primers, 25 out of 32 known (D. officinale×D. huoshanense) hybrids were correctly identified rendering a positive identification rate of 78.1%; while, 28 out of 31 known (D. huoshanense×D. officinale) hybrids were positively identified with a rate of 90.3%.
The authenticity of hybrids created by reciprocal crossing between Dendrobium officinale and D. huoshanense was not always apparent. To resolve the confusion, this study applied SSR markers to differentiate them among 63 F1 generation of the hybrids. Three out of 21 pairs of the SSR primers were found to have polymorphic bands, i.e., 14.3% of all, in the parental lines. By using these 3 pairs of primers, 25 out of 32 known (D. officinale×D. huoshanense) hybrids were correctly identified rendering a positive identification rate of 78.1%; while, 28 out of 31 known (D. huoshanense×D. officinale) hybrids were positively identified with a rate of 90.3%.
2017, 32(8): 874-879.
doi: 10.19303/j.issn.1008-0384.2017.08.013
Abstract:
Newly geminated buds of Dendrobium spp., Sanya Sunny, were used asexplants to induce bud clumps. Effects of culture media (i.e., Hyponex #1, Improvement #1, Improvement #2, and Improvement #3), phytohormones (i.e., 6-BA, NAA, and IBA), and sugar on the bud clump induction, propagation, and rooting of the plantlets were analyzed and optimized using anorthogonal experiment. The results showed that the major factors affecting the bud multiplication were medium, 6-BA, NAA, and sugar. Improvement #2 medium that contained 1.0 mg·L-1 of 6-BA, 0.2 mg·L-1 of NAA, 30.0 g·L-1 of sugar, 3.0 g·L-1 of agar powder and 3.6 g·L-1 of carrageenanprovided the best average propagation coefficient of 6.25 in 50 d among all tested media. For rooting, Improvement #3 with added NAA at 0.3 mg·L-1, IBA at 0.3 mg·L-1, AC at 0.5 g·L-1, banana puree at 100.0 g·L-1, sugar at 20.0 g·L-1, agar powder at 3.6 g·L-1 and carrageenan at 3.6 g·L-1 performed best with a 100.0% rooting rate. The survival rate of the plantlets in 60 d after transplanting from test tubes was 96.5%.
Newly geminated buds of Dendrobium spp., Sanya Sunny, were used asexplants to induce bud clumps. Effects of culture media (i.e., Hyponex #1, Improvement #1, Improvement #2, and Improvement #3), phytohormones (i.e., 6-BA, NAA, and IBA), and sugar on the bud clump induction, propagation, and rooting of the plantlets were analyzed and optimized using anorthogonal experiment. The results showed that the major factors affecting the bud multiplication were medium, 6-BA, NAA, and sugar. Improvement #2 medium that contained 1.0 mg·L-1 of 6-BA, 0.2 mg·L-1 of NAA, 30.0 g·L-1 of sugar, 3.0 g·L-1 of agar powder and 3.6 g·L-1 of carrageenanprovided the best average propagation coefficient of 6.25 in 50 d among all tested media. For rooting, Improvement #3 with added NAA at 0.3 mg·L-1, IBA at 0.3 mg·L-1, AC at 0.5 g·L-1, banana puree at 100.0 g·L-1, sugar at 20.0 g·L-1, agar powder at 3.6 g·L-1 and carrageenan at 3.6 g·L-1 performed best with a 100.0% rooting rate. The survival rate of the plantlets in 60 d after transplanting from test tubes was 96.5%.
2017, 32(8): 880-884.
doi: 10.19303/j.issn.1008-0384.2017.08.014
Abstract:
Using the commonly applied protein evaluation methods, the nutritionalqualityof proteins in the fruiting bodies of Flammulina velutipes cultivated under simulated microgravity was compared with that of the mushrooms grown under regular condition (control). It was found that 14 amino acids (AA)in the mushrooms under treatment were higher than control. The contents of total AA and essential AA were 151.2 g·kg-1 (i.e., 26.32% increase over control) and 81.7 g·kg-1 (i.e., 29.89% increase over control), respectively. Thus, the weightless condition effectively raised the essential AA of F. velutipes 2.82% over control, 8.71% over chicken protein, and 52.63% over the FAO/WHO reference standard. Six protein nutritional indices, including chemical score (CS), AA score (AAS), AA ratio coefficient (AARC), essential AA index (EAAI), biological value (BV) and nutritional index (NI), of the treatment samples were higher than those ofcontrol. Itappeared that the simulated microgravitycultivation increased the AA and protein qualityof F. velutipes fruiting bodies over the conventional method.
Using the commonly applied protein evaluation methods, the nutritionalqualityof proteins in the fruiting bodies of Flammulina velutipes cultivated under simulated microgravity was compared with that of the mushrooms grown under regular condition (control). It was found that 14 amino acids (AA)in the mushrooms under treatment were higher than control. The contents of total AA and essential AA were 151.2 g·kg-1 (i.e., 26.32% increase over control) and 81.7 g·kg-1 (i.e., 29.89% increase over control), respectively. Thus, the weightless condition effectively raised the essential AA of F. velutipes 2.82% over control, 8.71% over chicken protein, and 52.63% over the FAO/WHO reference standard. Six protein nutritional indices, including chemical score (CS), AA score (AAS), AA ratio coefficient (AARC), essential AA index (EAAI), biological value (BV) and nutritional index (NI), of the treatment samples were higher than those ofcontrol. Itappeared that the simulated microgravitycultivation increased the AA and protein qualityof F. velutipes fruiting bodies over the conventional method.
2017, 32(8): 885-890.
doi: 10.19303/j.issn.1008-0384.2017.08.015
Abstract:
Gas chromatography-mass spectrometry was employed for the investigation on the heterogeneity of metabolites in tomato plants infected by strains of Ralstonia solanacearum with varied pathogenicity. The plants were inoculated with the virulent FJAT-91 and/or avirulent FJAT-1458, as well as water as control. After 6, 24, 48, 72 and 96 h, metabolites in the plants were analyzed. The results showed the metabolites were mostly alcohols, esters, acids, aldehydes, pyridines, and alkanes. The compositional differences on the plants infected by different pathogenic strains were compared. It was found that the number of metabolite varieties in the plants varied by the treatments and time. The tomato plants treated by FJAT-91 for 48 h had 12 different metabolites; those treated by FJAT-1458 for 48 h, 18;those treated by both strains simultaneously for 96 h, 15;and, control treated for 72 or 96 h, 15. Dibutyl phthalate was the only metabolite present in all samples. n-hexadecanoic acid was absent in the plants inoculated with FJAT-91, but was detected at considerable amounts in those infected with FJAT-1458 or combined strains, and control. The discrepancy might be due to a variation in the immunology of the tomato plants. A principal component analysis on the metabolites revealed discernible differences among the tomato plants that received different treatments. It suggested that the viral infections could be identified by comparing the two most abundant metabolites in a plant.
Gas chromatography-mass spectrometry was employed for the investigation on the heterogeneity of metabolites in tomato plants infected by strains of Ralstonia solanacearum with varied pathogenicity. The plants were inoculated with the virulent FJAT-91 and/or avirulent FJAT-1458, as well as water as control. After 6, 24, 48, 72 and 96 h, metabolites in the plants were analyzed. The results showed the metabolites were mostly alcohols, esters, acids, aldehydes, pyridines, and alkanes. The compositional differences on the plants infected by different pathogenic strains were compared. It was found that the number of metabolite varieties in the plants varied by the treatments and time. The tomato plants treated by FJAT-91 for 48 h had 12 different metabolites; those treated by FJAT-1458 for 48 h, 18;those treated by both strains simultaneously for 96 h, 15;and, control treated for 72 or 96 h, 15. Dibutyl phthalate was the only metabolite present in all samples. n-hexadecanoic acid was absent in the plants inoculated with FJAT-91, but was detected at considerable amounts in those infected with FJAT-1458 or combined strains, and control. The discrepancy might be due to a variation in the immunology of the tomato plants. A principal component analysis on the metabolites revealed discernible differences among the tomato plants that received different treatments. It suggested that the viral infections could be identified by comparing the two most abundant metabolites in a plant.
2017, 32(8): 891-896.
doi: 10.19303/j.issn.1008-0384.2017.08.016
Abstract:
An aromatic oolong tea was used as the raw material to extract polysaccharides (TPs) and other constituents (TPs-A, TPs-B and TPs-C) using two processing methods. The chemical compositions, reducing power, and DPPH scavenging capacities of the extracts were determined. The results showed that the method applying 65% ethanol for pre-soaking followed by a water extraction (E-W) produced more TPs and less soluble protein than the process employing a water extraction before precipitation by 65% ethanol (W-E). E-W promoted the extractions of polyphenols, catechins, flavonoids and caffeine; while, W-E was less efficient in separating or concentrating the components. On the antioxidant activity, TPs obtained by both methods were significantly lower on the reducing power and DPPH scavenging capacity than those of TPs-A, TPs-B and TPs-C. The correlation analysis suggested that the antioxidant activity of TPs was significantly related to the contents of polyphenols, catechins, and particularly ester catechins in the extract.There was also a synergism among TPs, total flavonoid and caffeine. It appeared that as antioxidants, TPs exhibited significantly lower activity than the phenolic compounds in the tea.
An aromatic oolong tea was used as the raw material to extract polysaccharides (TPs) and other constituents (TPs-A, TPs-B and TPs-C) using two processing methods. The chemical compositions, reducing power, and DPPH scavenging capacities of the extracts were determined. The results showed that the method applying 65% ethanol for pre-soaking followed by a water extraction (E-W) produced more TPs and less soluble protein than the process employing a water extraction before precipitation by 65% ethanol (W-E). E-W promoted the extractions of polyphenols, catechins, flavonoids and caffeine; while, W-E was less efficient in separating or concentrating the components. On the antioxidant activity, TPs obtained by both methods were significantly lower on the reducing power and DPPH scavenging capacity than those of TPs-A, TPs-B and TPs-C. The correlation analysis suggested that the antioxidant activity of TPs was significantly related to the contents of polyphenols, catechins, and particularly ester catechins in the extract.There was also a synergism among TPs, total flavonoid and caffeine. It appeared that as antioxidants, TPs exhibited significantly lower activity than the phenolic compounds in the tea.
2017, 32(8): 897-904.
doi: 10.19303/j.issn.1008-0384.2017.08.017
Abstract:
The ultrasound-assisted extraction of flavonoids from leaves of Helianthus tuberosus L. was optimized. The extract obtained was evaluated for its antimicrobial activity.Based on the results from single-factor tests, the extraction conditions were further optimized using the response surface methodology. It was found that when 40% ethanol was applied as the solvent in a substrate:solvent ratio of 1:40 to extract the leaves for 20 min at 64℃, the yield and efficiency of the process could be maximized to reach a flavonoid extraction rate of 1.47 mg·g-1. The flavonoids obtained showed an excellent antibacterial activity on Escherichiacoli, but not on Staphylococcus aureus or Bacillus subtilis.
The ultrasound-assisted extraction of flavonoids from leaves of Helianthus tuberosus L. was optimized. The extract obtained was evaluated for its antimicrobial activity.Based on the results from single-factor tests, the extraction conditions were further optimized using the response surface methodology. It was found that when 40% ethanol was applied as the solvent in a substrate:solvent ratio of 1:40 to extract the leaves for 20 min at 64℃, the yield and efficiency of the process could be maximized to reach a flavonoid extraction rate of 1.47 mg·g-1. The flavonoids obtained showed an excellent antibacterial activity on Escherichiacoli, but not on Staphylococcus aureus or Bacillus subtilis.
2017, 32(8): 905-908.
doi: 10.19303/j.issn.1008-0384.2017.08.018
Abstract:
To determine the genetic injury on the root-tip cells of Vicia faba caused by lead in water, a micronucleus assay was conducted.The genotoxic effect of lead acetate in the concentrations ranging from 0.5 mg·L-1 to 30 mg·L-1 were monitored and pollution indices calculated. The results showed that micronuclei in the cells increased with increasing lead concentration. When the cells were exposed to lead acetate at the concentration of 30.0 mg·L-1 for 6 h, the number of micronuclei in the cells were significantly higher than that in control (P < 0.05).The pollution index was greater than 3.5, which was considered as a heavily polluted condition. Moreover, in the cases where the cells were treated with lead concentrations at 1.0-30.0 mg·L-1 for 12 h, similar results were observed at an extremely significant level(P < 0.01). At a same lead concentration, the 12 h treatments affected the cells significantly greater than the 6 h treatments. However, the difference was only at a significant level(P < 0.05) for 1 mg Pb·L-1. It seemed apparent that lead was absorbed to causegenetic injury to cells in the roots of V. faba, and the toxicity increased with dosage and time of the exposure.
To determine the genetic injury on the root-tip cells of Vicia faba caused by lead in water, a micronucleus assay was conducted.The genotoxic effect of lead acetate in the concentrations ranging from 0.5 mg·L-1 to 30 mg·L-1 were monitored and pollution indices calculated. The results showed that micronuclei in the cells increased with increasing lead concentration. When the cells were exposed to lead acetate at the concentration of 30.0 mg·L-1 for 6 h, the number of micronuclei in the cells were significantly higher than that in control (P < 0.05).The pollution index was greater than 3.5, which was considered as a heavily polluted condition. Moreover, in the cases where the cells were treated with lead concentrations at 1.0-30.0 mg·L-1 for 12 h, similar results were observed at an extremely significant level(P < 0.01). At a same lead concentration, the 12 h treatments affected the cells significantly greater than the 6 h treatments. However, the difference was only at a significant level(P < 0.05) for 1 mg Pb·L-1. It seemed apparent that lead was absorbed to causegenetic injury to cells in the roots of V. faba, and the toxicity increased with dosage and time of the exposure.
2017, 32(8): 917-920.
doi: 10.19303/j.issn.1008-0384.2017.08.020
Abstract:
Polysaccharidesin loaches were extracted by an ultrasound-assisted enzymatic method to determine the antioxidant activityusing an in vitro biochemical assay. The results showed that the polysaccharides had strong scavenging capacities toward hydroxyl radicals, superoxide anions, and hydrogen peroxide.Theremoval rate of the extract onhydroxyl radicals reached as high as 73.38%, while 88.22% on hydrogen peroxide and 57.99% onsuperoxide anions.Atotal antioxidant capacity of 8.42 units·mL-1 of the loach polysaccharides was considered high and desirable.
Polysaccharidesin loaches were extracted by an ultrasound-assisted enzymatic method to determine the antioxidant activityusing an in vitro biochemical assay. The results showed that the polysaccharides had strong scavenging capacities toward hydroxyl radicals, superoxide anions, and hydrogen peroxide.Theremoval rate of the extract onhydroxyl radicals reached as high as 73.38%, while 88.22% on hydrogen peroxide and 57.99% onsuperoxide anions.Atotal antioxidant capacity of 8.42 units·mL-1 of the loach polysaccharides was considered high and desirable.
2017, 32(8): 909-916.
doi: 10.19303/j.issn.1008-0384.2017.08.019
Abstract:
This article reviews the status of collecting, protecting and applying existing resource of tea germplasms, breeding new varieties and formulating objectives, compiling information on tea origins and suitability for processing, promoting business and new products, as well as searching for means to improve farmers' income for the tea industry in Fujian. The resource on germplasms for breeding is abundant in the province. There are more than 7,000 varieties collected and well preserved for studies and applications. The objectives set forth by the industry and government on tea breeding have undergone changes over the years, from early budding to high yield to excelled quality. To date, there have been 45 cultivars successfully bred locally. Among them, 35.6% were deemed suitable for making oolong, black or green teas, and 26.7% for white tea. This allowed a flexibility for tea processing and fresh leaves utilization. Each new cultivar was bred individually based on the conditions of cultivation, process technology and market demand. As a result, the plantation acreage and production of teas had rapidly increased in the past, supported by the optimized operations and marketing for the industry. Prospects in the areas of germplasm collection, breeding diversification and specialization, unique cultivar promotion, and processing technology development on teas are discussed.
This article reviews the status of collecting, protecting and applying existing resource of tea germplasms, breeding new varieties and formulating objectives, compiling information on tea origins and suitability for processing, promoting business and new products, as well as searching for means to improve farmers' income for the tea industry in Fujian. The resource on germplasms for breeding is abundant in the province. There are more than 7,000 varieties collected and well preserved for studies and applications. The objectives set forth by the industry and government on tea breeding have undergone changes over the years, from early budding to high yield to excelled quality. To date, there have been 45 cultivars successfully bred locally. Among them, 35.6% were deemed suitable for making oolong, black or green teas, and 26.7% for white tea. This allowed a flexibility for tea processing and fresh leaves utilization. Each new cultivar was bred individually based on the conditions of cultivation, process technology and market demand. As a result, the plantation acreage and production of teas had rapidly increased in the past, supported by the optimized operations and marketing for the industry. Prospects in the areas of germplasm collection, breeding diversification and specialization, unique cultivar promotion, and processing technology development on teas are discussed.
