2011 Vol. 2, No. 2
2011, 2(2): 97-100.
doi: 10.15886/j.cnki.rdswxb.2011.02.018
Abstract
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Abstract:
In our study,the head kidney tissue were used as materials,and in vivo treatment by phytohemagglutinin（PHA） and colchicines and Giemsa staining were performed,and the cytogenetics of E.trimaculatus were analyzed.The results indicated that the karyotypic formula of E.trimaculatus was 2n=48,48t,NF=48,no heteromorphic chromosome was detected.Based on the karyotypes of 23 species of groupers,the conclusion that 2n=48,48t,NF=48 was the primitive cytogenetic characterization of genus Epinephelus were proposed.
In our study,the head kidney tissue were used as materials,and in vivo treatment by phytohemagglutinin（PHA） and colchicines and Giemsa staining were performed,and the cytogenetics of E.trimaculatus were analyzed.The results indicated that the karyotypic formula of E.trimaculatus was 2n=48,48t,NF=48,no heteromorphic chromosome was detected.Based on the karyotypes of 23 species of groupers,the conclusion that 2n=48,48t,NF=48 was the primitive cytogenetic characterization of genus Epinephelus were proposed.
2011, 2(2): 101-106.
doi: 10.15886/j.cnki.rdswxb.2011.02.008
Abstract:
In order to determine varieties and contents of froctooligosaccharides in transgenic sugarcane transformed with 1-SST.Two saccharides determination methods using HPLC were established,one for determining the fructose,glucose and sucrose,the other for determining three Froctooligosaccharides（GF2,GF3 and GF4）.Saccharides of sugarcane were extracted with water,using a mobile phase composed of acetonitrile and water（87/13 V/V;75/25 V/V） at a flow rate of 1.0 mL·min-1 for separating,Hydersir NH2 column（4.6 mm×250 mm,5 μm） temperature was 30 ℃,the differential refraction detector was used to detect samples.The results indicated that the linear ranges of the six saccharides were 0.625—50 mg·mL-1,with a correlation coefficient exceeding 0.999 3.kestose GF2 in sugarcane transformed with the 1-SST gene were detected.Our methods can be used for accurate qualitative and quantitative determination of fructose,glucose and sucrose in sugarcane.
In order to determine varieties and contents of froctooligosaccharides in transgenic sugarcane transformed with 1-SST.Two saccharides determination methods using HPLC were established,one for determining the fructose,glucose and sucrose,the other for determining three Froctooligosaccharides（GF2,GF3 and GF4）.Saccharides of sugarcane were extracted with water,using a mobile phase composed of acetonitrile and water（87/13 V/V;75/25 V/V） at a flow rate of 1.0 mL·min-1 for separating,Hydersir NH2 column（4.6 mm×250 mm,5 μm） temperature was 30 ℃,the differential refraction detector was used to detect samples.The results indicated that the linear ranges of the six saccharides were 0.625—50 mg·mL-1,with a correlation coefficient exceeding 0.999 3.kestose GF2 in sugarcane transformed with the 1-SST gene were detected.Our methods can be used for accurate qualitative and quantitative determination of fructose,glucose and sucrose in sugarcane.
2011, 2(2): 107-112.
doi: 10.15886/j.cnki.rdswxb.2011.02.013
Abstract:
A cDNA fragments homologue of the known plant mitogen activated protein kinase（MAPKK,MKK,MEK） was selected by the suppression subtractive hybridization（SSH） library,and which can be induced by ethylene,and Rapid Amplification of cDNA Ends（RACE） were used to obtain the full length cDNA.Bioinformatic analysis results suggested that the gene belongs to a subfamily of plant MAPKK with a conserved region phosphorylation consensus sequence S/T*****S/T.Semi-quantitative RT-PCR results indicated that the expression of this gene increased after ethylene treatment,which suggested it may play an important role in the ethylene induction pathway.
A cDNA fragments homologue of the known plant mitogen activated protein kinase（MAPKK,MKK,MEK） was selected by the suppression subtractive hybridization（SSH） library,and which can be induced by ethylene,and Rapid Amplification of cDNA Ends（RACE） were used to obtain the full length cDNA.Bioinformatic analysis results suggested that the gene belongs to a subfamily of plant MAPKK with a conserved region phosphorylation consensus sequence S/T*****S/T.Semi-quantitative RT-PCR results indicated that the expression of this gene increased after ethylene treatment,which suggested it may play an important role in the ethylene induction pathway.
2011, 2(2): 113-116.
doi: 10.15886/j.cnki.rdswxb.2011.02.006
Abstract:
In the paper,a method for constructing dsRNA prokaryotic expression vector was introduced.The conserved region of PRSV-CP gene,which is 278 bp of 3'-end region,was used as the base,three hairpin structures of different lengths（861 bp,432 bp,279 bp） were cloned and constructed through one-step,zero-background ligation-independent cloning（OZ-LIC） method,and which were successively cloned into pSP73 prokaryotic expression vector after being digested with XhoⅠand ClaⅠ.Finally three recombinants of dsRNA prokaryotic expression plasmid of PRSV-CP gene were obtained.
In the paper,a method for constructing dsRNA prokaryotic expression vector was introduced.The conserved region of PRSV-CP gene,which is 278 bp of 3'-end region,was used as the base,three hairpin structures of different lengths（861 bp,432 bp,279 bp） were cloned and constructed through one-step,zero-background ligation-independent cloning（OZ-LIC） method,and which were successively cloned into pSP73 prokaryotic expression vector after being digested with XhoⅠand ClaⅠ.Finally three recombinants of dsRNA prokaryotic expression plasmid of PRSV-CP gene were obtained.
2011, 2(2): 117-122.
doi: 10.15886/j.cnki.rdswxb.2011.02.016
Abstract:
In order to identify the proteins in Hevea that interacted specifically with JAZ,the recombinant plasmid that expressed fusion protein of Hevea JAZ（HbJAZ1） and GST（Glutathione S-transferase） was constructed and transformed into Escherichia coli（E.coli） for prokaryotic expression and further purification.The results indicated that HbJAZ1-GST fusion protein was successfully expressed and purified,and the fusion proteins exist in pellet of bacterial lysis suspension.
In order to identify the proteins in Hevea that interacted specifically with JAZ,the recombinant plasmid that expressed fusion protein of Hevea JAZ（HbJAZ1） and GST（Glutathione S-transferase） was constructed and transformed into Escherichia coli（E.coli） for prokaryotic expression and further purification.The results indicated that HbJAZ1-GST fusion protein was successfully expressed and purified,and the fusion proteins exist in pellet of bacterial lysis suspension.
2011, 2(2): 123-128.
doi: 10.15886/j.cnki.rdswxb.2011.02.019
Abstract:
Microsatellite makers were used to evaluate genetic variations of Orange-Spotted Grouper（Epinephelus coioides） in the South China Sea wild population and the Hainan cultivation artificial propagation population,and 48 individuals in each stock were assessed at 19 SSR loci.The result showed that the wild population（WP） and cultivation artificial propagation population（CP）,the number of effective alleles（Ne） were 1.135 0—4.516 2 and 1.00 0—3.982 7 respectively;values of mean observed heterozygosity（Ho） were 0.7061 and 0.687 5 respectively;expected heterozygosity（He） was 0.480 3 and 0.421 5 respectively;polymorphic information content（PIC） were 0.547 5 and 0.493 0 respectively;genetic similarity coefficient（Gst）,genetic differentiation coefficient（Fst） and genetic distance（D） between WP and CP were 0.979 4,0.015 7 and 0.020 8 respectively.These genetic parameters suggested that the genetic variations in the South China Sea wild population of E.coioides were well conserved and the genetic differentiation and degradation were not obvious in Hainan cultivation artificial propagation population.
Microsatellite makers were used to evaluate genetic variations of Orange-Spotted Grouper（Epinephelus coioides） in the South China Sea wild population and the Hainan cultivation artificial propagation population,and 48 individuals in each stock were assessed at 19 SSR loci.The result showed that the wild population（WP） and cultivation artificial propagation population（CP）,the number of effective alleles（Ne） were 1.135 0—4.516 2 and 1.00 0—3.982 7 respectively;values of mean observed heterozygosity（Ho） were 0.7061 and 0.687 5 respectively;expected heterozygosity（He） was 0.480 3 and 0.421 5 respectively;polymorphic information content（PIC） were 0.547 5 and 0.493 0 respectively;genetic similarity coefficient（Gst）,genetic differentiation coefficient（Fst） and genetic distance（D） between WP and CP were 0.979 4,0.015 7 and 0.020 8 respectively.These genetic parameters suggested that the genetic variations in the South China Sea wild population of E.coioides were well conserved and the genetic differentiation and degradation were not obvious in Hainan cultivation artificial propagation population.
2011, 2(2): 129-132.
doi: 10.15886/j.cnki.rdswxb.2011.02.009
Abstract:
In our report,the chromosome was prepared using root tips of cassava variety SC6,the specific primers of MeEFⅠwere designed accoding to the complete cDNA sequence in GenBank,and in situ PCR was used to locate MeEFⅠin Manihot esculenta crantz genome.The results indicated that one or two signals were detected by in situ PCR on all phases of cell division;MEEFⅠgene located on the short arm of chromosome 10;the percent distances of the gene from centromere is 35.11.
In our report,the chromosome was prepared using root tips of cassava variety SC6,the specific primers of MeEFⅠwere designed accoding to the complete cDNA sequence in GenBank,and in situ PCR was used to locate MeEFⅠin Manihot esculenta crantz genome.The results indicated that one or two signals were detected by in situ PCR on all phases of cell division;MEEFⅠgene located on the short arm of chromosome 10;the percent distances of the gene from centromere is 35.11.
2011, 2(2): 133-137.
doi: 10.15886/j.cnki.rdswxb.2011.02.010
Abstract:
PCR were performed to clone LTP3 promoter from color cotton.Sequencing analysis results indicated that it contained 1 548 nt;the homology is 99% compared with the LTP3 reported by Liu,and only 4 nucleotides were changed.The LTP3（1548bp） promoter was fused with the E.coli.β-glucuronidase gene（GUS） and transformed into tobacco.The results suggested that GUS expressions under control of LTP3 promoter were detected only in the trichomes of transgenic tobacco leaves,while the positive control,contains Cauliflower mosaic Virus（CaMV） 35S promoter and GUS gene,was constitutively expressed,and there are no GUS expression in the negative control（untransformation plants）.The results indicated that LTP3 promoter can be used to express the heterologous genes in tobacco leaves.
PCR were performed to clone LTP3 promoter from color cotton.Sequencing analysis results indicated that it contained 1 548 nt;the homology is 99% compared with the LTP3 reported by Liu,and only 4 nucleotides were changed.The LTP3（1548bp） promoter was fused with the E.coli.β-glucuronidase gene（GUS） and transformed into tobacco.The results suggested that GUS expressions under control of LTP3 promoter were detected only in the trichomes of transgenic tobacco leaves,while the positive control,contains Cauliflower mosaic Virus（CaMV） 35S promoter and GUS gene,was constitutively expressed,and there are no GUS expression in the negative control（untransformation plants）.The results indicated that LTP3 promoter can be used to express the heterologous genes in tobacco leaves.
2011, 2(2): 138-142.
doi: 10.15886/j.cnki.rdswxb.2011.02.011
Abstract:
The effects of three template DNA extraction methods from different growth stages on transgenic sugarcane PCR detection were compared.The results indicated that the quality of DNA extracting from seedling leaves was better than that of mature plants and mature stage,and PCR detections were easier;DNA extracted from sugarcane leaves with CTAB method,SDS method and CTAB-SDS method were all suitable for PCR detection of resistant plants in seedling stage.The quality of DNA extracted with CTAB-SDS method was the best for PCR detection of resistant plants in mature stage.
The effects of three template DNA extraction methods from different growth stages on transgenic sugarcane PCR detection were compared.The results indicated that the quality of DNA extracting from seedling leaves was better than that of mature plants and mature stage,and PCR detections were easier;DNA extracted from sugarcane leaves with CTAB method,SDS method and CTAB-SDS method were all suitable for PCR detection of resistant plants in seedling stage.The quality of DNA extracted with CTAB-SDS method was the best for PCR detection of resistant plants in mature stage.
2011, 2(2): 143-147.
doi: 10.15886/j.cnki.rdswxb.2011.02.012
Abstract:
The young leaves of Pogostemon cablin were used as explants,and a large number of shoots were obtained through inducing callus.The shoots were transferred onto the multiplication media with different concentration of 6-BA,and onto the root-promoting media with different concentration of IBA.The optimum multiplication culture medium was obtained（MS +0.1 mg·L-1—0.2 mg·L-1 6-BA）,and the optimum root-promoting medium was obtained（1/2 MS +0.25 mg·L-1—1.0 mg·L-1 IBA）.When the height of the plantlets of Pogostemon cablin amounted to 5—6 cm,the survival percentage of plantlets Pogostemon cablin in the transplantation was 95%.
The young leaves of Pogostemon cablin were used as explants,and a large number of shoots were obtained through inducing callus.The shoots were transferred onto the multiplication media with different concentration of 6-BA,and onto the root-promoting media with different concentration of IBA.The optimum multiplication culture medium was obtained（MS +0.1 mg·L-1—0.2 mg·L-1 6-BA）,and the optimum root-promoting medium was obtained（1/2 MS +0.25 mg·L-1—1.0 mg·L-1 IBA）.When the height of the plantlets of Pogostemon cablin amounted to 5—6 cm,the survival percentage of plantlets Pogostemon cablin in the transplantation was 95%.
2011, 2(2): 148-152.
doi: 10.15886/j.cnki.rdswxb.2011.02.007
Abstract:
The TAS-ELISA and DAS-ELISA were established to detect the orchids suspected to be infected by virus mainly from main orchid planting in Hainan Province.The results suggested that the pathogen were Cymbidium mosaic virus（CymMV）,Odontolossum ring spot virus（ORSV） and their mixed infection;there were more orchids which were infected with CymMV than with ORSV.The method possesses high sensitivity,accuracy and specificity.
The TAS-ELISA and DAS-ELISA were established to detect the orchids suspected to be infected by virus mainly from main orchid planting in Hainan Province.The results suggested that the pathogen were Cymbidium mosaic virus（CymMV）,Odontolossum ring spot virus（ORSV） and their mixed infection;there were more orchids which were infected with CymMV than with ORSV.The method possesses high sensitivity,accuracy and specificity.
2011, 2(2): 153-156.
doi: 10.15886/j.cnki.rdswxb.2011.02.014
Abstract:
In order to find the best DNA extraction method of asparagus,improved SDS,CTAB and kit method were used to extract DNA from Asparagus young leaves and the DNA quality also was tested by UV spectrophotometer,agarose gel electrophoresis and PCR detection.The results showed that the quality of the DNA from the three DNA extraction methods fill the requirements for ISSR molecular markers and other assays.
In order to find the best DNA extraction method of asparagus,improved SDS,CTAB and kit method were used to extract DNA from Asparagus young leaves and the DNA quality also was tested by UV spectrophotometer,agarose gel electrophoresis and PCR detection.The results showed that the quality of the DNA from the three DNA extraction methods fill the requirements for ISSR molecular markers and other assays.
2011, 2(2): 157-163.
doi: 10.15886/j.cnki.rdswxb.2011.02.017
Abstract:
In our study,according to the Hoxc8 sequence of cow,the specific primers were designed,and the sequences of Hoxc8 exon-1（432 bp）and exon-2（273 bp）of normal and multi-thoracic vertebrae mongolia sheep were obtained（Genebank accession number: EU817489 and FJ905472）.Alignment results of them indicated that the sequences were conformity except a little difference in two sides of sequences.Hoxc8 exon-1 and exon-2 were aligned with other species and the results showed that compared with other mammals（human,dog,mouse,rat and chimpanzee）,the homology were above 96%（exon-1） and 91%（exon-2）;compared with zebra fish,the homology were 75.8% and 74%.
In our study,according to the Hoxc8 sequence of cow,the specific primers were designed,and the sequences of Hoxc8 exon-1（432 bp）and exon-2（273 bp）of normal and multi-thoracic vertebrae mongolia sheep were obtained（Genebank accession number: EU817489 and FJ905472）.Alignment results of them indicated that the sequences were conformity except a little difference in two sides of sequences.Hoxc8 exon-1 and exon-2 were aligned with other species and the results showed that compared with other mammals（human,dog,mouse,rat and chimpanzee）,the homology were above 96%（exon-1） and 91%（exon-2）;compared with zebra fish,the homology were 75.8% and 74%.
2011, 2(2): 164-166.
doi: 10.15886/j.cnki.rdswxb.2011.02.015
Abstract:
In order to promote the extraction of pectin from pineapple peel,the acid hyclrolyzed suspending solution of pineapple peel was treated with cellulose,and the conditions of enzymatic hydrolysis process were screened.The results indicated that the optimal conditions were as follows: enzymatic hydrolysis temperature was 55℃;enzymatic hydrolysis pH volume was pH 4.2;appending quantity of cellulose was 15.5 U·mL-1;and enzymatic hydrolysis time was 140 min,and under these conditions,the concentration of pectin in the enzymatic hyclrolyzed solution reached 7.4 g·L-1,which was 40.7% higher than that of the acid hyclrolyzed solution.
In order to promote the extraction of pectin from pineapple peel,the acid hyclrolyzed suspending solution of pineapple peel was treated with cellulose,and the conditions of enzymatic hydrolysis process were screened.The results indicated that the optimal conditions were as follows: enzymatic hydrolysis temperature was 55℃;enzymatic hydrolysis pH volume was pH 4.2;appending quantity of cellulose was 15.5 U·mL-1;and enzymatic hydrolysis time was 140 min,and under these conditions,the concentration of pectin in the enzymatic hyclrolyzed solution reached 7.4 g·L-1,which was 40.7% higher than that of the acid hyclrolyzed solution.
2011, 2(2): 167-171.
doi: 10.15886/j.cnki.rdswxb.2011.02.003
Abstract:
In our study,the soil fertility of Casuarinn equies forest,landscape shelterbelt and non-forest land in coastal area of Haikou were investigated and evaluated.The results indicated that the pH value is neutral;available nitrogen is medium level;available phosphorus and potassium are low;organic matter are low seriously;the index of landscape shelterbelt was much better than that of the other two forest;organic matter and moisture content have a significant correlation with other factors;the whole level of coastal area of Haikou is poor;organic matter content was the main factors that affected the forest soil fertility level.
In our study,the soil fertility of Casuarinn equies forest,landscape shelterbelt and non-forest land in coastal area of Haikou were investigated and evaluated.The results indicated that the pH value is neutral;available nitrogen is medium level;available phosphorus and potassium are low;organic matter are low seriously;the index of landscape shelterbelt was much better than that of the other two forest;organic matter and moisture content have a significant correlation with other factors;the whole level of coastal area of Haikou is poor;organic matter content was the main factors that affected the forest soil fertility level.
2011, 2(2): 172-177.
doi: 10.15886/j.cnki.rdswxb.2011.02.001
Abstract:
In this study,the route and sample plot investigation were performed to survey the species diversity of insects in Wugui mountain of Zhongshan urban nature reserve.After insect collection,the dry preserved specimens were prepared;some familiar insect species were identified according to the professional illustrated handbook;some insect species which could not be identified by the limited condition were then sent to insect experts for identification.Totally,40 thousand specimens were prepared,among which the moths catalogue of 286 species from 29 families which belong to Lepidoptera were firstly reported.
In this study,the route and sample plot investigation were performed to survey the species diversity of insects in Wugui mountain of Zhongshan urban nature reserve.After insect collection,the dry preserved specimens were prepared;some familiar insect species were identified according to the professional illustrated handbook;some insect species which could not be identified by the limited condition were then sent to insect experts for identification.Totally,40 thousand specimens were prepared,among which the moths catalogue of 286 species from 29 families which belong to Lepidoptera were firstly reported.
2011, 2(2): 178-186.
doi: 10.15886/j.cnki.rdswxb.2011.02.004
Abstract:
Based on the depth interviews and questionnaires on banana cooperatives in China,analytic hierarchy processes were performed to study the performance of banana cooperatives.The results indicated that the cooperatives can solve some difficulties and problems in the process of the development of banana industry,and improve the degree of scale and organization of banana industry,but the overall performance level is not high.
Based on the depth interviews and questionnaires on banana cooperatives in China,analytic hierarchy processes were performed to study the performance of banana cooperatives.The results indicated that the cooperatives can solve some difficulties and problems in the process of the development of banana industry,and improve the degree of scale and organization of banana industry,but the overall performance level is not high.
2011, 2(2): 187-192.
doi: 10.15886/j.cnki.rdswxb.2011.02.002
Abstract:
In the paper,the species of plant endogenous RNA and related gene silencing pathways and the constitute and function of main effectors in the silencing pathways were reviewed.In order to further explore the complexity and diversity of regulation process in plant gene expression,the differences in their function between miRNA and siRNA were discussed.The paper provides basic introduction for the utilization of plant endo signal pathway and will prompt the further research work in this field.
In the paper,the species of plant endogenous RNA and related gene silencing pathways and the constitute and function of main effectors in the silencing pathways were reviewed.In order to further explore the complexity and diversity of regulation process in plant gene expression,the differences in their function between miRNA and siRNA were discussed.The paper provides basic introduction for the utilization of plant endo signal pathway and will prompt the further research work in this field.
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