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2013 Vol. 28, No. 7

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Sequence Analysis of ITS-1 and Partial HSP70 Gene of Three Cryptocaryon irritans Strains
CHI Hong-shu, LIU Xiao-dong, KE Ling, GONG Hui
2013, 28(7): 625-629. doi: 10.19303/j.issn.1008-0384.2013.07.001
Abstract:
The ciliates witch isolated from Paralichthys olivaceus in Changle, Epinephelus awoara in Fuding and Pseudosciaena crocea in Sandu′ao Ningde were identified as Cryptocaryon irritans based on morphological observation.The genomic DNA of the ciliates witch was extracted, and the gene sequence of the internal transcribed spacer 1 (ITS-1) were amplified by PCR, then cloned, sequenced and analyzed.The results revealed that the ITS1nucleotide sequence of the three strains were exactly the same.Homology comparation indicated that they shared 100%identity with the gene sequence of PYH4.12strain (DQ270010) , Wakayama strain (AB608054) , Chiayi strain (AF490381) , and shared 90.62%-99.22% identity with other sequences in NCBI database, which confirmed that the three identified strain was Cryptocaryon irritans.Furthermore, partial HSP70gene sequence was amplified from the genomic DNA of the identified Cryptocaryon irritans by PCR, and only three base pairs difference were observed in the sequence among the three strains.This indicated that the Cryptocaryon irritans Strains gotten from different hosts, different farming models in Fujian were highly homologous.
cDNA Cloning and Sequencing for Precursor of Fujian Yellow Rabbit Follicle Stimulating Hormone β Subunit
SANG Lei, SUN Shi-kun, CHEN Dong-jin, CHEN Yan-feng, XIE Xi-ping
2013, 28(7): 630-633. doi: 10.19303/j.issn.1008-0384.2013.07.002
Abstract:
The total RNA of the Fujian yellow rabbit′s pituitary was used to amplify the cDNA for the precursor of its follicle stimulating hormoneβsubunit (FSHβ) by means of RT-PCR.The opening reading frame of 390bp was obtained.The fragments amplified were ligated with pMD18-T vector and transformed into DH5αcompetent cells.Randomly selected positive clones were sequenced and submitted to GenBank for verification (accession number: KC784943) .It was compared with homologous sequences of the mammalian animals, including human, cow, sheep, buffalo, and mouse.The sequence alignment showed the similarities ranging from 72.2%to 96.5%.The genetic evolution analysis on the gene indicated a fairly high degree of homology of the rabbit with the primates.The cDNA sequences of the Fujian yellow rabbit′s FSHβwere also compared with those of the Japanese white rabbit′s.Nucleotide mutations at C81Tand G219Awere found, but not on amino acids.
Molecular Identification and Rapid Detection of Pathogenic Plesiomonas shigelloides from Ctenopharyngodon idellus
ZHANG Xin-yan, FAN Hai-ping, CENG Zhan-zhuang
2013, 28(7): 634-638. doi: 10.19303/j.issn.1008-0384.2013.07.003
Abstract:
A Gram negative pathogenic bacterium named strain LCCiL90625NA was isolated from the liver samples of diseased Ctenopharyngodon idellus and was indentified by phylogenetic analysis of 16SrRNA.A phylogenetic tree was constructed based on 16SrDNA sequences of strain LcCiL90625NA and other related bacteria species in the GenBank.It revealed that strain LcCiL90625NA shared a high similarity of 99.7% with Plesiomonasshigelloides and formed a cluster with strain PIC3 (accession number GQ359957) .The isolated bacterium strain LcCiL90625NA was recognized as Plesiomus shigelloides.PCR primers were designed to amplity a 280-bp flagment from the Plesiomusshigelloides23SrRNA gene.The results indicated that this method might serve as an efficient tool for rapid and sensitive identification of Plesiomus shigelloides.
Development and Application of Ligase-dependent RT-PCR Assay for Detection of Avian Tembusu Virus
FU Guang-hua, HUANG Yu, CHENG Long-fei, CHEN Cui-teng, WAN Chun-he, CHEN Hong-mei, PENG Chun-xiang, SHI Shao-hua, LIN Jian-sheng
2013, 28(7): 639-643. doi: 10.19303/j.issn.1008-0384.2013.07.004
Abstract:
Based on the mechanism of ligase-dependent RT-PCR assay, we established RT-PCR assay to detect avian tembusu virus which still prevails in most poultry of China.The results showed that it would specifically detect tembusu viruses isolated from different avian, such as laying duck, meat duck, goose and hens.The sensitivity of this assay is equivalent to that of conventional RT-PCR.To test the reliability of ligase-dependent RT-PCR assay, six tissue samples collected from experimental animals and 140clinical samples collected from Fujian, Jiangxi, Zhejiang, and Guangdong province during 2011-2012were detected.All of the six tissue samples were positive, while only 20of the 140clinical samples were tested to be positive by this method.The above results indicated that the ligase-dependent RT-PCR assay was a sensitive, specific and rapid diagnosis method for detecting avian Tembusu virus.
Prokaryotic Expression of the NS1 Gene for Avian Tembusu Virus
PAN Yi-zhe, WAN Chun-he, HUANG Yu, CHENG Long-fei, FU Guang-hua, SHI Shao-hua, CHEN Hong-mei
2013, 28(7): 644-647. doi: 10.19303/j.issn.1008-0384.2013.07.005
Abstract:
A pair of primers were designed and synthesized according to the nanostructure protein (NS1) gene of avian Tenbusu virus WR strain available in GenBank.The NS1gene was amplified by RT-PCR and cloned into the prokaryotic expression vector pGEX-4T-1, then induced into E.coli with IPTG.The SDS-PAGE analysis of expressed protein demonstrated that the recombinant NS1protein of 62kD had got a high expression, which laid a good foundation for developing in molecular diagnosis and the function of the NS1gene for avian Tenbusu virus.
Effects of the Dietary Protein Levels on the Growth and Feed Utilization of Juvenile Lateolabrax japonicas
LIN Xing
2013, 28(7): 648-652. doi: 10.19303/j.issn.1008-0384.2013.07.006
Abstract:
To investigate the effect of the diet protein level on growth and feed utilization of juvenile Lateolabrax japonicus, five isoenergetic experimental diets containing 35.0%, 37.5%, 40.0%, 42.5% and 45.0% protein were made up respectively, whose protein source was fish meal.Juvenile Lateolabrax japonicus with initial average weight of (84.81±0.92) g took the feeding experiments for 75days for determination of the best protein level.The results showed that the relative weight gain rate, specific growth rate and feed conversion rate of the group of 39.85% were much higher than those of 34.76% and 37.54% groups (P<0.01) , but not significantly different from the 42.34% group (P>0.05) .Protein efficiency rate of the 39.85%group was also higher than those of the 34.76%, 42.34% and 45.03% groups (P<0.05) , while comparing with the 37.54% group, no significant difference was found (P>0.05) .In this experiment, based on quadratic model regression analysis of the specific growth rate and protein efficiency rate, growth index and feed utilization rate of Lateolabrax japonicus were best while the protein content of juvenile Lateolabrax japonicus feed was 38.87%-41.50%.
Varieties of Fishes and Growth Characteristics of Aristichthys nobilis in Shanmei Reservoir
SHUI Xing-yong, LIAO Jia-wei, CAI Yong-zong, ZHU Lun-zhang, WANG Yi-min, CHEN Jin-tian, ZHANG Hui-shan, HUANG Yong-chun
2013, 28(7): 653-658. doi: 10.19303/j.issn.1008-0384.2013.07.007
Abstract:
Thirty-seven species in 10fish families were found in Shanmei Reservoir.They included 22species of Cyprinidae, accounting for 59.46% of the total, and 4species of Bagridae, accounting for 10.81% of the total.There were also respectively 2species of Channidae and Anguillidae, and one species of Cichlaidae, Siluridae, Serranidae, and Adrianichthyidae.Specifically, the growth characteristics of Aristichthys nobilis found in the reservoir were observed: (a) its body weight (W) could be related to the length (L) as W=0.0269×L2.9132; (b) its body length to the scale size (R) as L=0.0129×R+3.5323; (c) its Von Bertalanffy growth equations were Lt= 107.2167×[1-e-0.1349 (t+0.1561) ]and Wt =22095.9566×[1-e-0.1349 (t+0.1561) ]3; (d) its growth inflection point occurred at 7.99-year-old; (e) its maximum body length, L∞, was 107.22cm; (f) its maximum body weight, W∞, reached 22.1kg;and, (g) the growth coefficients were k=0.1349and t0 =-0.1561.
Effects of Different Nitrogen Fertilizer Applications on Physiological Characteristics,Yield,and Quality of Dual-Purpose Peanut
WENG Bo-qi, ZHENG Xiang-li, ZHAO Ting, XU Guo-zhong, WANG Jun-hong
2013, 28(7): 659-663. doi: 10.19303/j.issn.1008-0384.2013.07.008
Abstract:
The field experiment was conducted with peanut‘shan G'as material, in the experimental station of Agricultural Ecology Institute, Fujian Academy of Agricultural Sciences to study the effects of N application amount on physiological characteristics, yield and quality of dual-purpose peanut.The results showed that different nitrogen fertilizer treatments increased chlorophyll content and photosynthetic rate in peanut leaves, with greater increasing extent at middle-later stage (pod filling stage) than that at the middle stage (pod setting stage) .Nitrogen application enhanced the activities of catlase (CAT) and the contents of soluble protein, but reduced malondialdeyde (MDA) content in peanut leaves, especially treated by N300and N450.Nitrogen application led to the highest yields of peanut and peanut vine, and reached the highest when applying N 300kg·hm-2 level.In addition, the application of nitrogen fertilizer also increased the contents of lysine and methionine, which were inadequate in the protein fractions of peanut kernel, to improve nutritional quality of peanut.
Analysis on Yield Components of Series CMS-FA Hybrid Rice Combinations Derived from Restorer Line Jinhui 3
CHOU Xiu-li, WANG Hong-fei, WANG Nai-yuan, LIANG Kang-jing
2013, 28(7): 664-669. doi: 10.19303/j.issn.1008-0384.2013.07.009
Abstract:
Relationship between yield and yield components of series CMS-FA hybrid rice combinations derived from restorer line Jinhui 3was studied.It aimed at detecting the yield potential of CMS-FA hybrid rice combinations crossbred with restorer line Jinhui 3.Using 7CMS-FA sterile lines as female parents and corresponding restorers Jinhui 3as male parents, 7crosses were made, along with Shanyou 63as the control, were used for the study.The results showed that series CMS-FA hybrid rice yield an average of 7 877.8kg·hm-2, the majority of combination of yield was significantly or very significantly higher than the control.The yield was significantly positively correlated with total grain number per plant (TGNPP) , and not significantly correlated with effective panicles per plant (PP) , seed-set percent (SSP) and 1000-grain-weight (TGW) .Yield was positively directly affected by TGNPP, and was negatively directly affected by PP and TGW.When using restorer line Jinhui 3to cross CMS-FA hybrid rice combinations, CMS-FA sterile line should be selected within a certain PP, TGW should not be too large as appropriate.
Effects of Different Culture Medium on Ophiopogon japonicus (L.F) Kew-Gawl. in vitro Culture
WAN Xue-feng, LAI Zhong-xiong, CHEN Jing-ying
2013, 28(7): 670-674. doi: 10.19303/j.issn.1008-0384.2013.07.010
Abstract:
The effects of NH+ 4 and NO3 levels and BA, NAA on vitro proliferation and rooting of Ophiopogon japonicus (L.F) Kew-Gawl were compared with vitro culture, Ophiopogon japonicus (L.F) Kew-Gawl to study the conditions of Ophiopogon japonicus (L.F) Kew-Gawl.The results showed that the coefficient of japonicus proliferation was 2.60under the optimum ratio of NH+ 4 to NO3 was 2.2∶1with BA.Further, the proliferation with BA was higher than that without BA at the same condition.In addition, the root proportions of different nitrogen conditions were also different.When the ratio of NH+ 4 to NO3 was 1.7∶1, the situation was the best for rooting without NAA.The trend of rooting was almost the same between with NAA and without NAA.But it was not rooting when the ratio of NH+ 4 to NO3 was 2∶1.
Optimization of Tissue Culture for Propagating Clustered Shoots of Phalaenopsis
ZHONG Hai-feng, ZHONG Huai-qin, HUANG Min-ling, WU Jian-she, YE Xiu-xian
2013, 28(7): 675-679. doi: 10.19303/j.issn.1008-0384.2013.07.011
Abstract:
Adventitious shoots induced from the peduncle auxiliary buds of Phalaenopsis‘Lauluns' were used for the study.The physical segmentation, placement mode and concentration of inorganic salt and organic additives all showed significant effects on the proliferation of Phalaenopsis with clustered shoots.Among the treatments, the greatest average number of inductive buds occurred when a bud was excised at the growth point and then grafted horizontally (PSP-3) .However, the bud's death rate was as high as 23.3%.In contrast, when the double buds were used, and followed by the horizontal grafting (PSP-6) , the budding was high, but the death rate was low.The inorganic salt concentration between 1/3MS to 1/2MS was found to be suitable for the propagating with strong seedling growth.Addition of coconut juice at 15% (v/v) level was desirable for the Phalaenopsis cluster buds proliferation, while 150g·L-1 potato juice or banana puree could be applied as low cost organic additives in the culture medium.
Breeding of Melon Rootstock“NANZHEN 1”
WU Yu-fen, ZHAO Yi-jie, CHEN Sheng, ZHANG Wei-guang
2013, 28(7): 680-684. doi: 10.19303/j.issn.1008-0384.2013.07.012
Abstract:
“NANZHEN 1”is a new variety melon rootstock hybrid derived from female parent 012and male parent 023.Its characteristics is highly fusarium wilt resistantance, good compatibility with melon, graft survival rate above 90%, more than 10% that of Lianjiang pumpkin rootstock.It is obvious to improve muskmelon yield, comparing with own root seedlings.It is little influence on melon fruit quality, basic keeps the original flavor and quality characteristics of the melon.
Regional Test of a New Oolong Tea Variety ‘Ruixiang'
YOU Xiao-mei, CHEN Zhi-hui, ZHONG Qiu-sheng, LIN Zheng-he, CHEN Rong-bing, CHEN Chang-song
2013, 28(7): 685-689. doi: 10.19303/j.issn.1008-0384.2013.07.013
Abstract:
With Huangdan as the control species (CK) , the national regional test of the new tea variety‘Ruixiang' was conducted for 6years in Guangdong, Guangxi and Hunan provinces.The investigating indices of Ruixiang in the regional test referred to the rate of survival, growth potential, yield of fresh leaves, quality of made tea and its environmental resistance.Resultsshowed that Ruixiang grew well, had the high rate of survival, strong environmental resistance, high yield of fresh leaves, and good quality of made tea.It could be an middle-maturing, excellent-quality and high-yielding tea variety, which was of good adaptability in plantation, popularization and application in Guangdong, Guangxi and Hunan provinces and others Oolong tea regions.
Biological Characteristics of the Plant Vaccin Strain FJAT-9290 Against Fusarium Wilt
ZHAN Hong, XIAO Rong-feng, LIU Bo
2013, 28(7): 690-696. doi: 10.19303/j.issn.1008-0384.2013.07.014
Abstract:
The biological characteristics of plant vaccin strain (nonpathogenic Fusarium oxysporum strain FJAT9290) were investigated by compare the effects of incubation time, and nutritional conditions on its growth with a pathogenic strain as a control.The results showed that there was no significant difference of the mycelium dry weight between strain FJAT-9290with the pathogenic strain FJAT-282, the sporulation of strain FJAT-9290was 1.47×106 cfu·mL-1 on the 6th day, while the strain FJAT-282is 108.33×106 cfu·mL-1, and changes of pH values had no significant effect on the culture growth.Furthermore, the concentration of reducing sugar in culture filtrate of strain FJAT-9290decreased faster than strain FJAT-282;starch was conducive to the growth of mycelium of strain FJAT-9290, glucose was conducive to its sporulation, xylose was not conducive to the growth of mycelium of strain FJAT-282, fructose was conducive to its sporulation;organic nitrogen was benefit to the growth of mycelium, ammonium salt was not conducive to the growth of hypha, but also to the sporulation.In addition, nitrogen sources exercised a greater influence on colony morphology.
Effect of Temperature on the Growth and Antifungal Activity of Fusarium-wilt-disease Biocontrol Bacterium Strain FJAT-4
GE Ci-bin, LIU Bo, XIAO Rong-feng, ZHU Yu-jing, TANG Jian-yang
2013, 28(7): 697-704. doi: 10.19303/j.issn.1008-0384.2013.07.015
Abstract:
To characterize the effect of temperature on the growth and antifungal activity of biological control strain Bacillus licheniformis FJAT-4, the growth response of strain FJAT-4at different temperature, the effect of concentration and temperature on the antifungal activity of the strain FJAT-4against Fusarium oxysporum strain FJAT-135were studied by culturing the strain on plate culture media, liquid media and culturing the strain FJAT135on the media contained strain FJAT-4.The results showed that the growth is inhibited and the growth curve is liner type under 20℃, but the growth return to normal and the growth curve is logarithmic type over 25℃, and the optimal temperature is 35-40℃.The cultured liquids of strain FJAT-4raised at different temperature and its 10 times serial dilution inhibited the growth of the strain FJAT-135effectively, and the differences among the inhibitions were far less than the differences among the concentrations of the cultured liquid.There were significant differences among effects of temperatures on the inhibitions of FJAT-4on FJAT-135, the inhibitory effect was the weakest at 15℃, and obvious at 20-30℃ and the highest at 35℃.Therefore, there were significant relationship between the temperatures and the growth and antifungal activity of strain FJAT-4, and the optimal temperature for the growth and antifungal activity was 35-40℃.
The Prey Preference and Functional Response of Adult Female Amblyseius eharai to the Eggs of Bemisia tabaci
JI Jie, LIN Tao, ZHANG Yan-xuan, CHEN Xia, SUN Li, LIN Jian-zhen
2013, 28(7): 705-708. doi: 10.19303/j.issn.1008-0384.2013.07.016
Abstract:
In the present study, the prey preference of the predatory mite Amblyseius eharai to the new eggs (yellow) and old eggs (brown) of Bemisia tabaci was examined.Our results showed that the predation efficiency of adult female A.eharai to the new eggs of B.tabaci is higher, the mean daily consumption are 8.6 (new) and 2.2 (old) eggs/day when the new/old eggs was supplied only, and the A.eharai preyed the new ones when the new/ old eggs was supplied simultaneously.Furthermore, the consumption by the adult female A.eharai increased along with the increasing of the predation density, and the predation efficiency increased along with the temperatures (16℃to 28℃) .The predation efficiency of A.eharai at 32℃ was higher than that at 20℃or 16℃, and was lower than that at 24℃ or 28℃.In addition, the highest rate of discovery of prey (a = 0.8054) and the shortest handling time (Th =0.0227) were obtained at 28℃.
Tolerance of Lactic Acid Bacteria Strains for Phytoprotein Yoghurt Fermentation to Simulated Gastrointestinal Environments
LIU Yun, CAO Yi, LIU Bo, LIU Dan-ying, CHEN Qian-qian
2013, 28(7): 709-713. doi: 10.19303/j.issn.1008-0384.2013.07.017
Abstract:
Previous studies have revealed that Lactobacillus plantarum FJAT-7926and Lactobacillus casei FJAT7928are promising fermentation agents for phytoprotein yoghurt beverage.The aim of our study was to examine the tolerance differences of the two strains under artificial gastric juice with different pH values and artificial intestinal juice containing different concentrates of bile salt and trypsin.The results showed that both of them have high tolerance responses to low pH value.When incubated in artificial gastric juice with pH3.0for 4h, survival of each tested strain was over 1×108cfu·mL-1 with a survival rate of 56.06%and 55.65%, respectively.However, the survival of them declined to 1×105cfu·mL-1 when incubated in the juice with pH2.0for 2h, and remained at concentrations of 3.67×102 cfu·mL-1 and 2.30×103 cfu·mL-1 in incubation for 4h.Our results suggest that both of the tested strains have great tolerance to the artificial intestinal juice.Though the survival rate of FJAT7928and FJAT-7926decreased as the concentration of bile salt increased, the concentrates of the both strains kept in the same order of magnitude.After 4hincubation in the artificial intestinal juice containing 0.3% or 0.5% bile salt, the survival rates for FJAT-7926and FJAT-7928were more than 70% and 40%, respectirely.The above indicated that both FJAT-7926and FJAT-7928have high tolerance to simulated gastrointestinal environments and both of them have broad prospects in the application of fermentation of phytoprotein yoghurt beverage.
Evaluation on in vitro Antioxidant Activity of Southern Wild Grape Wines
LI Wei-xin, GUO Yan-bo, HE Zhi-gang, LIN Xiao-zi, LIANG Zhang-cheng, REN Xiang-yun
2013, 28(7): 714-717. doi: 10.19303/j.issn.1008-0384.2013.07.018
Abstract:
Using Vitamin C solution as control in different concentrations, “brier grape wine”and“Gui-pu No1 wine”were studied with UV spectrophotometric assay on their scavenging activity to DPPH·, ·OH and reducing activity to potassium ferricyanide.The results showed that scavenging activity of two grape wines increased with the wine concentration.The scavenging activity to DPPH·in two wines were 2.75and 3.14time equivalent of it in Vc (2.0 mg·mL-1) , respectively, and to·OH were 6.98and 7.86time equivalent in 1.0 mg·mL-1, respectively.The scavenging speeds to free radicals were following the order in“Gui-pu No1wine”>“brier grape wine”>VC.There was no obvious difference between two wines in reducing activities to potassium ferricyanide but were higher in both wins than in Vc (0.5mg·mL-1) .
Pesticide Residue and Safety of Teas from Fujian
LIN Qiu, ZHANG Ju-de, SU De-sen, YAO Qing-hua, YAN Sun-an
2013, 28(7): 718-721. doi: 10.19303/j.issn.1008-0384.2013.07.019
Abstract:
Residue of 18pesticides in 55teas from Fujian were determined using GC-MS/MS and LC-MS/MS.Quantities of the residual pesticides were compared to the recommended limits, such as MRL, EMRL and/or ADI, to assess the product safety.Eleven samples, including black and oolong tea, had detectable amounts of prohibited pesticides.Some unregistered pesticides were also present in 26samples.Bifenthrin was the most frequently detected pesticides among all.The safety assessment indicated that only the level of imidacloprid residue in a certain tea samples was lower than MRL.And, in general, the daily pesticide ingestion from average tea drinking was within the safety limits set forth by ADI.
Study the Determination of Cellulose,Hemicellulose and Lignin in feed by FIBERTEC 2010 Semi-automatic Fiber Analyzer
LIU Wen-jing, PAN Wei, REN Li-hua
2013, 28(7): 722-726. doi: 10.19303/j.issn.1008-0384.2013.07.020
Abstract:
Van Soest method is commonly applied for fiber analysis worldwide, even though its measurement is known to be imprecise.Using an FIBERTEC2010Semi-automatic Fiber Analyzer, we determined the contents of cellulose, hemicellulose and lignin in feed to compare with the results obtained by the Van Soest method.The new method showed a variation coefficient of RSD less than 4% with a significantly improved degree of precision.This article presents the modified methodology as well as a formula for result calculation.