Acta Agriculturae Zhejiangensis

Establishment and optimization of AFLP system for Coix

2014, 26(1): 0-19.

Abstract ( 437 )

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By taking 30 samples of Coix species as testing materials， the genetic diversity and genetic relationship was analyzed using AFLP molecular markers technology. The optimal analysis system of AFLP has been established by studying the key factors of conventional process such as gene extraction， restriction digestion and link up， pre-amplication， selective amplification， electrophoresis and siliver staining detection. The result showed that the genome DNA was digested more completely by using two-step method compared with one-step method. In each step， the enzyme reaction time was 4 h added with 15 min for enzyme inactivation. The connection product should be diluted for 10 times and the pre-amplification product should be diluted for 50 times to get a better result. With application of the established system， 14 selective amplification primers with better polymorphism were selected out from 64 primers.

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Nitric oxide involved in defense responses of Oryza meyeriana to rice bacterial blight

2014, 26(1): 0-6.

Abstract ( 540 )

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Oryza meyeriana， a wild rice species， is highly resistant to rice bacterial blight （BB）， but the resistance mechanism is still unknown. Nitric oxide （NO） is an important signaling molecule that plays a pivotal role in plant disease resistance responses. However， little study has focused on whether NO is involved in BB resistance of O. meyeriana. In this study， the effects of BB pathogen inoculation on leaf lesion， NO content， NO subcellular location and xylem ultrastructure were studied in O. meyeriana and a susceptible rice cultivar Nipponbare. Pathogen infection caused yellow necrotic lesions in Nipponbare and brown apoptotic lesion in O. meyeriana， and the lesion length was much longer in Nipponbare than in the wild rice. No difference was found in NO content in Nipponbare after pathogen inoculation. However， NO content was prominently increased by pathogen inoculation in the wild rice， and it was further discovered that NO was largely accumulated in xylem cell wall. In addition， electron microscopic observations showed that BB pathogen inoculation induced the thickening of xylem cell wall in O. meyeriana but not in Nipponbare. Based on these results， we suggested that NO played an important role in the BB resistance of O. meyeriana， and the role might involve the thickening of xylem cell wall that could inhibit further invasion of the pathogen.

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Analysis and mapping of a dwarf and narrow leaf rice mutant

2014, 26(1): 0-13.

Abstract ( 344 )

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A dwarf and narrow leaf （dnl1） mutant was isolated by ethylmethylsulfonate （EMS） treating for a japonica rice variety Xiushui 09 dry seeds，it could be inherited stably after several selfing. Compared with its wild type Xiushui 09， the mutant shows dwarf， narrow leaf and more tillers. Genetic analysis showed that the traits of the dnl1 mutants were controlled by one pair of recessive nuclear gene. With map\|based cloning approach， Dnl1 gene was mapped between SSR marker RM17478 and RM17488 on the chromosome 4， a 260 kb region. Sequencing and blast analysis indicated that Dnl1 is an allele to the gene Nal1.

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Identification of the freshness of Litopenaeus vannamei with smell

2014, 26(1): 0-25.

Abstract ( 301 )

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Freshness of Litopenaeus vannamei changed， which gave out unpleasant smell， and its smell was respectively detected with the electronic nose and HS\|SPME\|GC\|MS， combined with the sensory evaluation and TVBN. The results showed that the output of five sensors of electronic nose， which was consisted of six sensors， presenting regular change. The volatile components in prawn were extracted by solid phase micro\|extraction （SPME） and determined by gas chromatography\|mass spectrometry （GC\|MS）.The volatile components in prawn were nitrogenous and sulfur compounds， alcohols and inflammable gas. Compared the change rule of output of each sensor of electronic nose with the detected results of volatile components， the two test results were better consistent. It has proved that the electronic nose could be tried to assess freshness of prawn.

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Development and evaluation of RT\|LAMP for the rapid detection of goose\|origin Tembusu virus

2014, 26(1): 0-33.

Abstract ( 296 )

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In this study， a RT\|LAMP （reverse transcription loop\|mediated isothermal amplification） assay was developed to rapidly detect and diagnose goose tembusu virus （GTMUV）. Using Primer Explorer V4 software， a series of specific primers were designed based on the nucleotide sequences of GTMUV NS5 gene available in GenBank. These primers targeted 6 distinct conserved sequences according to the conserved region of NS5 gene. The results showed the RT\|LAMP method specifically amplified GTMUV but not other common avian disease RNA viruses， and the optimum reaction temperature and time were verified to be 60℃ and 60 min. Compared with other methods， this method was 100 times more sensitive than the conventional PCR. Meanwhile， it did not require complicated thermal cycling steps and denaturation， with the addition of SYBR GreenⅠ dye in the reaction mixture， and the results could be differentiated by naked eyes. Thus， the results proved that RT\|LAMP was a reliable， accurate， simple and practical method for the detection of avian Tembusu virus in the field.

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Sequence cloning and prokaryotic expression of omp31 gene in swine Brucella

2014, 26(1): 0-28.

Abstract ( 383 )

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In this paper， the swine Brucella genome was used as the template to amplify the omp31 gene sequence and its sequence characteristics and recombinant expression through E. coli prokaryotic expression system were also invetigated. The results showed the ORF sequence of omp31 gene included 723 bp， and it encoded 240 amino acids. The recombinant protein Omp31 was successfully expressed by E. coli prokaryotic expression system. The recombinant protein was purified by GST\|tag affinity column， and the molucelar was consistent with the expected 553 KD.

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Effect of probiotics on water quality and bacterial community in shrimp pools at later stage of cultivation

WANG Xin;WU Yi\|fei;YAO Xiao\|hong;LIU Yong;SUN Hong;TANG Jiang\|wu*

2014, 26(1): 0-47.

Abstract ( 334 )

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The research of probiotics for aquatic animals is increasing with the demand of aquacultures sustainable development. To evaluate the effect of new developed probiotcs MP\|1 and MP\|2 and investigate their water purifying mechanism， experiments were conducted in shrimp pools at the later stage of cultivation. Water samples were sampled from the shrimp pools to determine the CODMn， ammonia， nitrite， nitrate and total nitrogen every week， and the bacterial community structure in shrimp pools were analyzed by PCR\|DGGE. Results showed the contents of nitrite in shrimp pools were all below 001 mg·L-1 after a week when the probotics were added. The contents of ammonia in shrimp pools kept within 026 mg·L-1 to 033 mg·L-1 and 035 mg·L-1 to 048 mg·L-1 after two weeks when probiotics MP\|1 and MP\|2 were added. The contents of nirate and CODMn remained statistical unchanged in the experiments. And the total nitrogen was found a removal rate over 341%. PCR\|DGGE showed the main bacterial populations and bacterial community structure changed significantly. The results suggested that probiotcs MP\|1 and MP\|2 could significantly improve the water quality while they were used in shrimp pools. It might be a result of the change in autochthonal bacterial community caused by probiotics.

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2014, 26(1): 0-39.

Abstract ( 432 )

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Porcine circovirus 2 （PCV2） is the primary causative factor of postweaning multisystemic wasting syndrome （PMWS） in swine. The first three open reading frames have been described while ORF4 is the fourth ORF in PCV2 genome and its function is still unknown. In this study， ORF4 gene was cloned into the vector of pGEX\|4T\|1 to construct recombinant plasmid pGEX\|4T\|ORF4. The recombinant plasmids confirmed by sequencing were transformed into BL21 competent strain and induced by IPTG. The purified GST\|ORF4 protein was subcutaneously injected into the rabbits for preparation of the anti\|GST\|ORF4 antibody. Blood was collected four weeks later and the titer of polyclonal antibody was assessed by indirect enzyme\|linked immune sorbent assay （ELISA）. The results showed that the GST\|ORF4 protein was successfully expressed by SDS\|PAGE analysis and could be specifically detected with resulting polyclonal antibodies against GST\|ORF4 protein by Western blot. The ELISA result indicated that the titer of the antibody was over 1∶12 800 and the antibody can be used for further functional study of ORF4 which may facilitate understanding the pathogenesis of PCV2 and interaction between PCV2 and host.

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Comparison of photosynthetic characteristics of different origin pepper seedlings under low temperature and poor light

LYU Xiao\|han;CHAI Wei\|guo

2014, 26(1): 0-53.

Abstract ( 418 )

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Under different low temperature and poor light treatment， the photosynthetic characteristics of four different origin pepper were studied. The results showed that with falling temperature and light intensity， photosynthetic parameters and chlorophyll fluorescence parameters of four different origin pepper substantially decreased. These results indicated that low temperature and poor light affected the photosynthesis of pepper seedlings by directly reducing the activity of the photosynthetic tissue and destroying PSⅡ reaction center. Besides， material 20805 originated in Turkey showed better low light utilization ability and stronger photochemical efficiency， material 07\|1\|1\|0\|1\|2 originated in the Yangtze River Delta region of China was the worst. It could be speculated that under low temperature and poor light， differences in photosynthetic characteristics of pepper seedlings might be due to the difference in the climate of pepper origin.

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Comparative analysis of endogenous hormones during dormant bud development of Chinese herbaceous peony

YUAN Yan\|bo;WANG Li\|hui;YU Xiao\|nan*

2014, 26(1): 0-60.

Abstract ( 291 )

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The growth of flower buds of plants is closely related to the endogenous hormones. This paper examined the buds of Paeonia cultivar Da Fugui in the period of dormancy. The content variation of the GA3， IAA， ZT， ABA in different periods of dormant bud development of Chinese herbaceous peony were measured by using the liquid chromatography technique. The results showed that ABA content started to increase on November 20th， reached the peak on December 10th; GA3 content increased obviously on December 10th and decreased on December 30th; ZT content had a similar trend to that of GA3. However， the IAA content was relatively low with little fluctuation in the dormant period. GA3/ABA and ZT/ABA values decreased with the deepening of dormancy and increased after the release of dormancy. The study indicated that GA3 and ZT can promote the release of dormancy， which has the theoretical guiding significance for the release of dormancy with the application of exogenous hormones.

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2014, 26(1): 0-66.

Abstract ( 341 )

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The experiment with different cultivation methods was carried out to study the effect of straw bagged with net cultivation method on yield， quality and development of purple yam （also named purple Dioscorea alata L.）. The results showed that the tuber numbers per plant of purple yam cultivated in straw bagged with net can be appropriately increased， at the same time the tuber was also relatively large， resulting in obvious yield increasing. The purple yam tuber cultivated in straw bagged with net had the relatively elongated shape， and avoided underground pest damage and injury in the growth process and harvest. So it had a good commodity with beautiful shape， smooth， integrity; in addition， straw cultivation is beneficial to improve nutritious and healthy compositions， for example， yam saponin， soluble sugar， polysaccharide， mucin polysaccharide and so on. Straw bagged with net cultivation method also belongs to the straw cultivation， though it reduced the yam saponin and mucin content， its nutritious and healthy compositions were still high. The straw bagged with net cultivation method can improve commodity and quality of the purple yam.

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Study on methods for RNA extraction from Phellinus baumii

GUO YAN\|cui;XIONG Yi;ZHANG Zhi

2014, 26(1): 0-99.

Abstract ( 311 )

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In order to obtain an ideal total RNA extraction method of Phellinus baumii， total RNA of Phellinus baumii was extracted with 3 methods including Trizol method， CTAB method and RNA prep pure Plant Kit method. The results showed that all these methods can extract total RNA from Phellinus baumii， however， the Trizol method was the best and had high integrity and purity. The band of 28S and 18S was clear. The value of A260/A280 was 196 and the RNA yield was 60464 μg·g-1. Thus， the RNA extracted by Trizol method could be used for RT\|PCR， gene cloning and gene expression and so on.

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Quantitative analysis of catechins in local registered tea cultivars in Hubei Province

JIN Xiao\|fang;JIA Shang\|zhi;SHI Ya\|ya;GONG Zi\|ming*

2014, 26(1): 0-71.

Abstract ( 378 )

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The contents of 8 main catechins in 12 local registered tea cultivars in Hubei Province were determined by high performance liquid chromatography （HPLC）. The results showed that the variation coefficients of catechins contents ranged from 1973% to 6717%. No significant trend was found within non\|esterified catechins， while contents of ester catechins decreased in the order of epigallocatechin gallate（EGCG）> gallocatechin gallate（GCG）> epicatechin gallate（ECG）> catechin gallate（CG）. According to the cluster analysis based on catechin quality index， the varieties Echa 1， Echa 8 and Echa 10 were suitable for the production of high quality green tea. Wufeng 212， Wufeng 310， Echa 3 and Echa 6 with high EGCG level could be used for the development of special tea drinks. Echa 2 contained high EGC and total catechin level. Therefore， it was of great potential for the breeding and production of black tea.

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In vitro toxicity and field control effect of six fungicides to Colletotrichum gloeosporioides causing citrus anthracnose

PU Zhan\|xyu;HUANG Zhen\|dong;HU Xiu\|rong;LI Hong\|ye;*

2014, 26(1): 0-126.

Abstract ( 362 )

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In order to obtain fungicides with high efficiency， low toxicity， and less pollution in controlling citrus anthracnose， which caused by Colletotrichum gloeosporioides Penz.， the toxicity of pyraclostrobin， azoxystrobin， prochloraz， pyraclostrobin·me\|tiram， thiophanate\|methyl and iprovalicarb to a strain of C. gloeosporioides was respectively tested by mycelium growth rate and spore germination. The results showed that the effective dose to reduce mycelial growth of C. gloeosporioides by 50% （EC50 values） were 081， 258， 0066， 497， 155515 and 23798 μg·mL-1， whereas the EC50 values of spore germination to pyraclostrobin， azoxystrobin， prochloraz and pyraclostrobin·me\|tiram were 093， 1434， 447 and 529 μg·mL-1. Field experiments showed that 95% of above citrus anthracnose could be inhibited by azoxystrobin， prochloraz， iprovalicarb and pyraclostrobin·me\|tiram with the recommended concentrations on citrus anthracnose. Conclusively， azoxystrobin， prochloraz， iprovalicarb and pyraclostrobin·me\|tiram are effective in controlling citrus anthracnose.

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Study on virus\|free and rapid propagation techniques of Zhuangyuanhong grape

YANG Guang;JIN Gui\|hua;DONG Jun;ZHANG Qing;GONG Na

2014, 26(1): 0-76.

Abstract ( 346 )

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Zhuangyuanhong grape was used as experimental material， the shoot tips of stem apexes were selected as explants， which reproduced rapidly through directly inducing cluster buds， then the methods of heat treatment and tip culture were combined to induce virus\|free grape. The results showed that shoots induction rate in medium 1/2MS was higher than MS. The best inducing effects was found in the medium of 1/2 MS+20 mg·L-1 6\|BA+01 mg·L-1 IBA. Rootless plantlets inoculated in the medium of 1/2MS+05 mg·L-1 IBA+02 mg·L-1 NAA grew quickly and the growth of adventitious root was good， which were ready for transplanting. The optimal heat treatment time was 10 days which was beneficial for growth and virus elimination of rootless plantlets. After the heat treatment， 100% target viruses in the rootless plantlets were taken off.

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Effects of exogenous bisulfite on photosynthetic pigments content and fruit growth of Red Fuji apple under different climate conditions

CHEN Ping\|zhao;SANG Zheng\|lin;HE Mei;HUANG Xian\|min;ZHANG Ze\|jun;WANG Rui

2014, 26(1): 0-83.

Abstract ( 336 )

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Spraying 5 mmol·L-1 NaHSO3 and KHSO3 on the 10\|11 a Red Fuji apple trees was carried out under high temperature， strong sunlight and arid condition or low temperature， poor sunlight and rainy condition to investigate the effect of bisulfate on the photosynthetic pigments content and fruit growth of Red Fuji apple， and water was used as the control. The results showed that the experimental groups were superior to the control significantly， and the effect of KHSO3 was better than that of NaHSO3. It was also concluded that the photosynthetic pigments under the low temperature， poor sunlight and rainy condition were higher than that under high temperature， strong sunlight and arid condition in both experimental group and the control obviously， but the fruit growth changed to the contrary.

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Construction of rapid propagation system for Orostachys fimbriatus

LI Qing\|wei;HOU Dian\|ming;LIANG Ming\|qin;LIU Rong\|ning

2014, 26(1): 0-88.

Abstract ( 291 )

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Studies on the multiple shoots induction， propagation and rooting of Orostachys fimbriatus plantlet in vitro were conducted to construct a rapid propagation system for Orostachys fimbriatus. The results were as follows: the proliferation multiple of the stem was 1169 when cultured with the MS+10mg·L-1 6\|BA+02mg·L -1 IBA+20g·L -1 sucrose medium. Leaves inoculated on MS+15mg·L -1 6\|BA+01mg·L-1 IBA+30g·L-1 sucrose medium could regenerate small adventitious buds well， and the number of regenerated bud per leaf was 971. The vigorous proliferous shoots were inoculated on 1/2 MS+02mg·L-1 IBA+005mg·L-1 NAA medium for rooting， and the rooting rate was 100%， the root number was 123 per shoot.

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Knock out deletion and functional analysis of glucose transportation system in Xanthomonas campestris pv. campestris

MA Chen;LIANG Ru\|bing;LIU Jian\|hua*

2014, 26(1): 0-104.

Abstract ( 481 )

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Three glucose transporters and two membrane\|bound glucose dehydrogenases were knocked out in the phytopathogenic bacterium Xanthomonas campestris pv. campestris 8004. The five mutants and Xcc8004 had little difference in their basic metabolisms， such as cell growth curve， extra\|cellular polysaccharide production and cellulases activities when cultured in nutrition\|rich medium. However， the deletion of sodium/glucose transporter gene （XC_2460） influenced the cell growth when glucose was used as carbon source，but its extra\|cellular glucose yield in carboxymethyl cellulose degradation test was about 167\|fold higher than those of other strains. These results demonstrated the hypothesis that the extracellular glucose concentration from cellulose could be increased significantly by block of glucose influx transport system without affecting basic cell metabolism is feasible.

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N\|terminal domain of VPg encoded by Wheat yellow mosaic virus is involved in the interaction with the nucleolar protein Fibrillarin

BIAN Jing;XIANG Rong;SUN Li\|ying;*;CHEN Jian\|ping;*

2014, 26(1): 0-94.

Abstract ( 496 )

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Wheat yellow mosaic virus （WYMV） is a member of the genus Bymovirus in the family Potyviridae. WYMV has two single\|stranded positive sense RNA genomes which produce 10 viral proteins. Among those proteins， viral genome\|linked protein （VPg）， which binds to the 5′ end of the viral RNA genome， is predicted to contain a nuclear localization signal （NLS） and nuclear export signal （NES） in the N\| and C\|terminal portions， respectively. WYMV（Wheat yellow mosaic virus） VPg fused to C\|terminus of enhanced green florescent protein （eGFP\|VPg） exclusively accumulated in the nucleus of epidermal cells of Nicotiana benthamiana plants. Fibrillarin， a nucleolus resident protein， was introduced into eYFP（n） plasmid. To investigate the regions of the VPg domain binding fibrillarin， a series of VPg mutant and fibrillarin performed BiFC assay. The result display VPg localized and interact with fibrillarin in the nucleus. NLS but not NES is required for interact with fibrillarin.

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Detection of Strawberry mottle virus and Strawberry mild yellow edge virus with multiplex RT\|PCR based on internal control

WANG Hong;WANG Jing\|jing;ZHANG Ke\|li;CHENG Zhuo\|min

2014, 26(1): 0-109.

Abstract ( 410 )

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Strawberry mottle virus（SMoV） and Strawberry mild yellow edge virus（SMYEV）， which are found world\|wide， can cause serious crop losses. Total RNA for reverse transcriptase\|polymerase chain reaction （RT\|PCR） was extracted from strawberry with CTAB method. A multiplex RT\|PCR assay was used for simultaneous detecting SMoV and SMYEV based on ndh B gene as the internal control. Both field plants and test\|tube plantlets of strawberries were detected effectively.

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Sensitivity to several types of insecticides in field populations of Spodoptera litura in Zhejiang Province

2014, 26(1): 0-116.

Abstract ( 442 )

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To characterize the developmental trend of resistance to insecticides of oriental leafworm moth （OLM）， Spodoptera litura， in Zhejiang province， the sensitivities to 11 insecticides of OLMs collected from Sanmen and Xiaoshan were studied and compared. Significant differences were shown between the data of OLMs from the two cities， i.e. the acquired sensitivities to 11 insecticides of OLMs from Sanmen were higher than those of OLMs from Xiaoshan. Refering to the survival rate of OLMs after treating at discriminating doses， together with the ratio of insecticidal LC99 to field recommended concentration， it could be deduced that OLMs in Sanmen was still sensitive to chlorantraniliprole， chlorfenapyr; sensitivities to indoxacarb， emamectin benzoate， chlorfluazuron， diafenthiuron， spinosad have declined; and resistances to tebufenozide， spinetoram and beta\|cypermethrin have developed. While the OLMs in Xiaoshan were sensitive to chlorantraniliprole， chlorfenapyr and spinosad; sensitivity to emamectin benzoate， diafenthiuron， chlorfluazuron， spinetoram， indoxacarb， abamection had declined， and resistances to beta\|cypermethrin have developed. Therefore， resistance levels of OLMs to most of the insecticides monitored were increasing in Zhejiang province， and beta\|cypermethrin， abamection， tebufenozide that OLMs had developed resistance should be stopped using for one or two years， and the rest still can be applied to control OLMs by the methods of limiting the frequency of spraying， rotation or mixture of different kinds insecticides in order to delay the development of resistance.

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Successive impacts of different temperatures on the growth， development and reproduction of Cyrtorhinus lividipennis over three generations

2014, 26(1): 0-121.

Abstract ( 529 )

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Cyrtorhinus lividipennis is a key predator of the major rice pest the brown planthopper （BPH）， Nilaparvata lugens. Biological control of BPH by C. lividipennis could be much more effective and feasible by improving the techniques of artificial breeding and field\|releasing. In this study， the successive effects of different temperatures （19℃， 23℃， 27℃and 31℃） on the growth， development and reproduction of C. lividipennis were determined over three generations in the laboratory. The results showed that the female nymph duration was significantly shortened and the number of offspring decreased as generations developed at 19℃， the female adult longevity was shortened and the number of offspring reduced at 31℃; However， no obvious change on nymphal survival rate， female nymphal duration， female fresh weight， adult longevity and fecundity was found at other tested temperatures. Our results indicated the development and reproduction of C. lividipennis could be suppressed at 19℃ and 31℃，which could provide a theoretical basis for artificial breeding and field\|releasing of C. lividipennis.

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Detection of Lily symptomless virus by real\|time RT\|PCR

2014, 26(1): 0-130.

Abstract ( 370 )

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Lily symptomless virus （LSV） is one of the main viruses infecting Liliaceae plants. In the study， a real\|time fluorescent RT\|PCR method for LSV detection was established with specific primers and probe designed from the conservative sequences in coat protein genes of LSV isolates. Specificity study showed that typical amplification curves could be obtained for two LSV\|positive samples， but not for Lily mottle virus， Tobacco ringspot virus and Tomato ringspot virus\|positive samples. Sensitivity of this method was 10 times higher than that of regular RT\|PCR. With this method， 53 batches of LSV were intercepted in 67 batches of lily imported from Holland since June 2012 by Ningbo Entry\|Exit Inspection and Quarantine Bureau， and the detection rate was 93%.

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Simultaneous detection for residue of imidacloprid and nitenpyram in cowpea using HPLC method

LIU Jing\|kun;WU Chun\|yuan;DENG Xiao;CHEN Shan\|shan;LI Qin\|fen;*

2014, 26(1): 0-134.

Abstract ( 530 )

PDF (470KB) ( 1064 )

The maximum absorption wavelength of imidacloprid and nitenpyram detected by UV spectrophotometer were 270 nm and 246 nm. To get optimum retention time and separation， the mobile phase should be acetonitrile and water （V/V， 3∶7）. Standard curves of two pesticides were developed， which fit a linear regression ranging from 005 to 5 μg·mL-1. After extracted by homogenate with acetonitrile and purified by C18 SPE column， the recovery rates of the two pesticides were 75%～110% at the levels of 01mg·kg-1， 05mg·kg-1 and 15mg·kg-1 with a RSD less than 5%， which showed a good accuracy and stability. This method with a limit of quantification （LOQ） of 001μg·g-1 could be used in the practical detections.

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Establishment of an NS1\|ELISA for detection of antibody against Tembusu virus and initial application

2014, 26(1): 0-140.

Abstract ( 299 )

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In order to establish a NS1\| ELISA for rapid detection of antibodies against duck Tembusu virus（DTV）， a series of studies were conducted. Firstly， the non\|structural protein NS1 gene of goose Tembusu virus JS804 strain was inserted into the prokaryotic vector pET32a， then pET32a\|NS1 were transformed into Escherichia coli BL12（DE3）and the combination protein NS1 （His\|NS1） was obtained with the induction of IPTG. The fusion protein was purified by extracting the inclusion bodies with urea， and the purified protein was used as coated antigen. Based on the best working condition， the indirect ELISA method（NS1\|ELISA） to detect the antibody of duck TVD was developed. The optimized working condition included: the coating antigen of purified DTV\|NS1 protein was 1925μg·well-1， the best package condition was 37℃ for 1 h and then over night at 4℃， the best dilution of testing serum was 1∶200， the best closure time was 1 h， the dilution of HRP conjugated anti\|duck lgG was 1∶1 000，the chromogenic time was 10 min， and the cut off\|value judging negative or positive was 0346（D450）. The specific tests showed that there were no cross\|reaction to the anti\|sera against avian influenza virus， Newcastle disease virus， infectious bronchitis virus and DTV， which indicated that the NS1\|ELISA was specific to anti\|sera against DTV. The intra\|plate and inter\|plate demonstrated that the coefficient of maximum variation was 67％and 82％ respectively， which showed the method had good stability. A total of 81 samples from affected ducks were tested and 43 samples were positive detected by the NS1\|ELISA， While 48 samples showed positive detected by the E\|ELISA， which showed the method had a high sensitivity. The results revealed the NS1\|ELISA could be used for laboratory diagnosis and sera\|survey for duck Tembusu virus infection.

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Expression and purification of the novel Resilin\|R5 fusion protein in Escherichia coli

MAO Zheng\|zheng;LIU Yong;SUN Yan;ZHOU Man;LIU Zi\|duo;*;TANG Jiang\|wu;*

2014, 26(1): 0-147.

Abstract ( 292 )

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A fusion protein Resilin\|R5 which was expected to have a superiority of high elasticity and silicon deposition was designed and expressed in Escherichia coli. At the same time， a facile purification method was built to obtain the new fusion protein. Firstly， the recombinant resilin\|r5 gene， which constructed by fusing the diatoms R5 peptide gene sequences to the 3’ end of Drosophila melanogaster resilin gene sequences， was synthesized after a rare codon optimization. Then， the new gene was inserted into the prokaryotic expression vector pET28a. The expression vector was transformed into E. coli BL21（DE3）pLysS strain， and the transformant was cultured in an auto\|induced medium to get resilin\|r5 gene expressed. At last， the His\|tagged recombinant fusion protein was purified by‘salting\|out and heatingmethod and Ni ion column affinity chromatography. The efficient expression of the fusion Resilin\|R5 reached 120 mg per liter culture. A solid foundation was built for characterization and innovative applications of the fusion protein Resilin\|R5 which has high elasticity and silicon deposition ability.

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The density and storage of soil organic carbon in Zhejiang Province

2014, 26(1): 0-153.

Abstract ( 356 )

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Soil is the core of terrestrial ecosystem. Investigations into soil carbon （C） pool would clarify the role of soil in terrestrial ecosystem C cycle and global change. In the present study， 145 soil profile data collected from 2009 to 2012 were analyzed by soil type and GIS method to estimate the soil C pool in Zhejiang Province. The total amount of soil organic carbon （SOC） in Zhejiang Province was about 081～083 Pg， and the average soil carbon density was 808～837 kg·m-2. The SOC density varied within soil types. The lowest SOC density was shown as a southeast strip from Jinqu Basin to Taizhou\|Wenzhou costal area， and the SOC density increased gradually outward to the north， west and south. Finally， the advantages and disadvantages of soiltype and GIS\|estimation method in SOC evaluation were compared.

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Study on the combination of county cultivated land demand in social transformation period

TIAN Qiu\|xia;ZHANG Ren\|zhi*

2014, 26(1): 0-176.

Abstract ( 465 )

PDF (694KB) ( 959 )

The twenty\|first century was in the rapid social transformation period. Whether the forecast on the county cultivated land demand was reasonable or not was related to the degrees of regional and global cultivated land protection pressure. Taking Jiajiang County as an example， the food safety law， GM(1， 1) model and time series forecasting method were used to forecast the cultivated land demand for Jiajiang County. And then combining the natural， economic and social situation， as well as the present land utilization situation， three kinds of prediction results were endowed with weights by means of AHP method， finally cultivated land demand in Jiajiang County was calculated by using fixed weight coefficient prediction. The results showed that the demand for cultivated land in 2020 and 2030 would be 14 49359 and 14 28945 hm2. The method could accurately investigate the demand for cultivated land in a certain year， taking natural， social， economic factors into account.

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Phosphorus status and its leaching loss risks in the soils of Chinese hickory orchards in Linan City， Zhejiang Province

2014, 26(1): 0-158.

Abstract ( 294 )

PDF (780KB) ( 1140 )

In this paper， 50 top soil samples （0\|20 cm） were collected from Chinese hickory （Carya cathayensis Sarg.）orchards located at Daoshi and Tuankou towns of Linan City， Zhejiang Province， to assess the potential risk of phosphorus （P） leaching induced by long\|term fertilizer application. Soil analysis showed that soil Olsen\|P had a great spatial variability， being 0\| 893 mg·kg-1. Analysis of correlations between Olsen\|P and CaCl2\|P showed that the critical values （change\|point） of soil Olsen\|P in Daoshi and Tuankou towns were 193 and 293 mg·kg-1， respectively. Soil Olsen\|P exceeding the critical values accounted for 30% and 45% of all the investigated orchards, respectively, in Daoshi and Tuankou towns.

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Study on the microbial diversity of primary Korean pine forests and artificial forests

XIONG Yi;GUO Yan\|cui;ZHANG Zhi

2014, 26(1): 0-164.

Abstract ( 289 )

PDF (570KB) ( 1038 )

By the use of BIOLOG microplate method， the soil microbial diversity characteristics of primary Korean pine forests and man\|made forest in Liangshui National Nature Reserve， Lesser khingan were studied. The results showed that the soil microbial AWCD values of the forest\|type varied with soil depth and seasons: in the surface and 0\|10 cm layer of soil， it was the highest in Korean pine forest and Tilia amurensis forest; in the 10\|20 cm layer of soil， it was the highest in Xingan larch forest in spring， summer and autumn， while it was the highest in Tilia amurensis forest in winter. The soil microbial diversity indexes of all soil layers were the highest in Tilia amurensis forest， and were significantly higher than those of man\|made forest. So we concluded that the soil microbial diversity of primary Korean pine forest was richer than that of the others.

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Surface structure characteristics of barbecue bamboo charcoal and its adsorption behavior of ammonium from aqueous solution

YUAN Ju\|hong;HU Mian\|hao;*;QIN Hui\|yuan

2014, 26(1): 0-170.

Abstract ( 377 )

PDF (954KB) ( 1045 )

The surface structure characteristics of barbecue bamboo charcoal was analyzed using X\|ray diffraction （XRD）， scanning electron microscope and X\|ray energy dispersive spectrometer （SEM\|EDS）， and Fourier transform infrared spectrometer （FTIR）. The adsorption behavior of ammonium onto bamboo charcoal was also investigated by adsorption experiments. The results showed that the barbecue bamboo charcoal was dominated with micro\|pore， and contained various oxygen\|functional groups （\|OH，\|COOH）， methylene， trans\|isoolefine， aromatic and aliphatic compounds. Ammonium adsorption equilibrium was reached within about 80 min， and the adsorption capacity of barbecue bamboo charcoal tended to increase with elevated ammonium concentration in solutions. The Freundlich isotherm equation could well describe the isothermal adsorption behavior of ammonium on barbecue bamboo charcoal. The ΔG of the equation is negative， while ΔH is positive， indicating that the adsorption reaction of ammonia nitrogen is spontaneous. The pseudo\|second\|order model could effectively explain the adsorption kinetics. Pore diffusion， surface adsorption and intra\|particle diffusion would affect the adsorption process.

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Effect of high fructose syrup content on the glass transition temperature of linden honey

ZHOU Guo\|yan;ZHANG Jian\|jun;LI Xuan\|chen

2014, 26(1): 0-180.

Abstract ( 391 )

PDF (408KB) ( 991 )

Making an effective detection method on honey adulteration detection was of important practical significance on monitoring the honey industry. In order to achieve the rapid detection of honey mixed with high fruit syrup， the glass transition temperature Tg of linden honey mixed with different contents of high fructose syrup was determined by differential scanning calorimetry （DSC）. The results showed that the pure linden honey and high fructose syrup had significantly different glass transition temperature， whose Tg values were -5545℃ and 6086℃， respectively. When the content of high fructose syrup was lower than 10%， the Tg values of the adulterated honey and pure linden honey was too close to distinguish. When high fructose syrup content was higher （20%， 40%， 80%）， there was a good linear relationship between high fruit syrup quality score and the Tg values of doping honey. In conclusion， the identification of the contents of the doped high linden honey based on the differential scanning calorimetry is feasible.

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Studies on the conditions of solid\|state fermentation for cellulase production by Trichoderma viride and cellulase immobilization

2014, 26(1): 0-193.

Abstract ( 279 )

PDF (1119KB) ( 1015 )

Using wheat bran and corn straw as raw materials， the influence of the ratio of wheat bran to corn straw， fermentation temperature and time， moisture content， initial pH value， nitrogen concentration， on cellulose production by solid\|state fermentation of Trichoderma viride were studied in this paper. Based on optimization of solid\|state fermentation by single\|factor test， orthogonal test was made. The results showed that the optimal solid\|state fermentation conditions for cellulase were listed as follows：the ratio of wheat bran to corn straw powder was 4∶1，nitrogen concentration was 2%， culture temperature was 28℃， fermentation time was 72 h， initial pH value was 5， inoculum level was 10% and moisture content （based on solid fermentation substrate） was 175%. In this condition，maximum cellulase activity was 1 1325 U·g-1，which was increased 849% than 6124 U·g-1 in initial fermentation condition. Cellulase was immobilized by embedding， cross\|linking and embed\|cross\|linking method， in which embed\|cross\|linking method was the best one.

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Improvement of tobacco quality by flavored proteinase degradation

XIAO Ming\|li;YANG Qing;LIN Rui\|feng;PU Yuan\|zhu;*;BAO Xiu\|ping;LI Xian

2014, 26(1): 0-185.

Abstract ( 332 )

PDF (1373KB) ( 1009 )

In order to improve the quality of upper tobacco leaves， proteins in tobacco leaves were degraded by flavored proteinase， and the physical structure， free amino acids， aroma component of the processed tobacco leaves were compared with the control. The protein content decreased by 191% after proteinase degradation， and the texture of tobacco leaves became loose. The sensory characters were improved due to increased aroma， lightened miscellaneous odors， lowered irritation， and cleaner aftertaste. Especially， the bitterness and pungent taste was obviously eliminated. Besides， the content of free amino acid and aroma significantly increased. The above results indicated that flavored proteinase degradation was effective to improve the smoking quality of upper tobacco leaves.

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Comparison on biogas production of different biogas digesters

XU Qing\|xian;GUAN Xue\|fang;LIN Bi\|fen;QIAN Lei;LIN Bin;*

2014, 26(1): 0-199.

Abstract ( 297 )

PDF (722KB) ( 1080 )

The traditional biogas digester， biogas digester of glass fiber reinforced plastic（GFRP） using solar energy heating system and intelligent biogas digester in Xin\|xing pig farm and Yong\|sheng pig farm in Fujian Province were compared for biogas production. The index of feed of the three different technology biogas digesters was close and HRT was 10 d. The fermentation temperature changed with climate temperature， COD levels at the entrance and exit of biogas digesters and biogas production rate of biogas digesters were compared and analyzed in one year. The result showed that， firstly in the same conditions， the fermentation temperature of GFRP biogas digester was 085℃ higher than that of traditional biogas digester， the biogas production rate of up\|flow GFRP biogas digester， baffling GFRP biogas digester and traditional biogas digester was 074， 044 and 035 m3·m-3·d-1 respectively. Secondly， the biogas production rate of GFRP biogas digester using solar energy heating system was 074 m3·m-3·d-1， which was higher than that of GFRP biogas digester without solar energy heating system. Thirdly， the biogas production rate of intelligent biogas digester was 093 m3·m-3·d-1， which was higher than that of GFRP biogas digester using solar energy heating system.

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A design of the feeding control system for calf

JIN Mi\|xiao;LI Ya\|min*;ZENG Li\|hua

2014, 26(1): 0-209.

Abstract ( 333 )

PDF (445KB) ( 1046 )

In order to realize the scientific feeding of calves and to improve the economic benefits， a calf automatic feeding control system was designed. Taking PC as management platform， this system was based on cow information management technology and wireless communication technology， an 8\|bit CMOS microcontroller STC89C52， temperature sensor DS1820， and MPX2100 type pressure sensor as the core hardware. It solved the scientific control problem of feeding temperature and the amount of milk， and realized the process from cow identification to precise feeding.

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Simulation of soil tillage process based on ANSYS

ZHAI Li\|xin;JI Chang\|ying;*;DING Qi\|shuo;YU Yin\|mei

2014, 26(1): 0-215.

Abstract ( 411 )

PDF (1275KB) ( 1018 )

In order to verify the feasibility of data simulation， the tillage experiment was carried out and the stress in the advancing direction of plough was measured. Based on ANSYS， soil tillage process was simulated， and the overall distribution of stress within deformation zone was studied as well as the changes of tillage resistance. It was shown that the highest stress toward the heading direction occurred in the soil deformation zone of the maximum curvature of plough. During the tillage process， the tillage resistance increased first and then reduced slightly， and finally reached stable.

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Establishment of dynamic evaluation indicator system of poverty alleviation in rural area—Taking Lanzhou for example

ZHANG Xiao\|ning;FU Ying*;MA Yan\|ling

2014, 26(1): 0-234.

Abstract ( 317 )

PDF (817KB) ( 992 )

The scientific and reasonable evaluation system is an essential element for successful poverty alleviation，but the available poverty alleviation evaluation system has many flaws. Therefore， this paper used AHP\|Delphi method to rebuild the dynamic evaluation system in the perspective of achieving sustainable development and anti\|poverty from four aspects: policy relevance， efficiency， alleviation effects and sustainable development abilities， and carried out the dynamic evaluation and research by taking Lanzhou City for example. The results showed that poverty alleviation in Lanzhou was good and the evaluation index system was feasible.

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Study on the farmer\|initiated contacts for the rural public goods—a test of two theories

LUO Fang;CHEN Li\|jun;TIAN Miao;WANG Qing;ZHOU Ye\|wang

2014, 26(1): 0-221.

Abstract ( 307 )

PDF (651KB) ( 995 )

The farmer\|initiated contact with all levels of government agencies or organization for the rural public goods is one of the effective patterns of demand preference expression. The rural public goods need is about social well\|being which is generally indicated by household income and education. There are two different classic models about the relationship research of perceived needs and social well\|being， namely parabolic theory and hierarchy theory of needs. The two theories were tested with the Hubei Province farmer data， and it was concluded that parabolic model was proved to be a much better predictor of the sample.

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The production risk of Chinese beekeeper and the mechanism of disaster compensation

XI Gui\|ping;ZHAO Zhi\|jun;WU Li\|nan;CHEN Ma\|lin

2014, 26(1): 0-228.

Abstract ( 360 )

PDF (817KB) ( 1033 )

There are many risks during the course of production to Chinese beekeeper， such as natural disaster， pesticide poisoning， bee diseases and epidemics， stealing， traffic accident and so on. The incidence rate of beekeeping risk is very high， which leads to severe damage. Jiangshan city， as the name of The home of bee in China， did useful researches and trial in the relief safeguard of beekeeping in order to improve the resistance risk ability of the industry. The specific practice， operating mechanism， comparison of the advantages and disadvantages of the two disaster compensation models， which is participating in the risk salvation of beekeeping in Zhejiang province and comprehensive insurance of beekeeping， were analyzed. In the end， some policy proposals were put forward， which could promote the risk salvation measure actively， strengthen government support to insurance company of beekeeping and the construction of the insurance legislation， explore policy agricultural insurance system of beekeeping， and popularize the disaster compensation in Zhejiang province to other areas in China.

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Adjustment of Taiwan agricultural policy and its implications to rural land scale operation in China Mainland

HUANG Yan\|ting;WU Yu

2014, 26(1): 0-240.

Abstract ( 298 )

PDF (734KB) ( 950 )

In order to get rid of the agricultural crisis， Taiwan had implemented a series of agricultural policy from the 1970s. But the primary measures failed to grasp the main point， so policy changed its course in the late 1970s and early 1980s as developing scale management of farmland in focus， which finally achieved remarkable results. The following measures included helping farmers to purchase farmland， promoting the common management， cooperation management and commission management， implementing land consolidation policy， stimulating land lease and so on. These policies have a profound theoretical basis and, therefore have an important referential significance to rural land scale operation in China Mainland.

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The impact of livelihood capital on the life satisfaction of urban migrant workers in Hangzhou

SU Fei;PANG Ling\|feng*;MA Li\|sha

2014, 26(1): 0-246.

Abstract ( 322 )

PDF (764KB) ( 1059 )

The lack of various livelihood capitals has become one of the biggest obstacles to migrant workers urban lives. Therefore improving the livelihood capital of migrant workers has become the foundation and key to eliminating poverty of migrant workers and to improve their life satisfaction. This paper established a dual logistic regression model and analyzed the impact of livelihood capital on the life satisfaction of urban migrant workers in Hangzhou City. The results showed that:①the livelihood capitals of migrant workers in Hangzhou has spatial differences， the value of livelihood capitals of suburb areas is the highest， followed by the central area， then the outer suburbs;② migrant workers have lower conversion capacity of all kinds of livelihood capitals， with the consideration of the lack of human capital， weak physical capital， insufficient financial capital， and limited social capital;③social capital and human capital are the most important factors to affect the life satisfaction of migrant workers in Hangzhou City. At last， some suggestions are proposed in the hope of providing some references to achieve the sustainable livelihoods of urban migrant workers in Hangzhou City.

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Causes and countermeasures of urban\|rural fringe environmental problems—Huaxin Town， Shanghai as a case

LI Jin\|wen

2014, 26(1): 0-253.

Abstract ( 350 )

PDF (1758KB) ( 1010 )

Due to the unique geographical location， socio\|economic and population structure， environmental problems in urban\|rural fringe differed from those in the traditional countryside， where agriculture was the dominant industry， and the local residents were the majority of population. As industries were transferred to urban\|rural fringe from center city， the total population and the proportion of migrant workers increased greatly， and the industrial pollution load in these area also became much higher than in traditional countryside. Compared with traditional countryside， domestic wastewater and refuse in rural\|urban fringe were much higher， and accordingly increased the difficulty of environmental management. The present paper took Huaxin Town， a typical rural\|urban fringe in Shanghai as a case， to analyze the causes of environmental problems in this area from both industrial development and industry\|worker migration. Contiguous stretches were suggested to solve the environmental problems in the rural\|urban fringe.

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Banana shape recognition based on elliptic Fourier descriptor

HU Meng-han;DONG Qing-li;*;LIU Bao-lin;ZHANG Chong-yang;YE Fei

2014, 26(1): 200-205.

Abstract ( 618 )

PDF (836KB) ( 1241 )

In this paper， a method based on computer vision and elliptic Fourier descriptor （EFD） was developed to recognize the banana shape. The number of banana with shapes of the curved， slightly curved， end-straight and straight was 107， and the bananas were previously indentified by the experts. Subsequently， the boundary chain codes of the pre-processed images of these bananas were obtained for extracting the elliptic Fourier coefficients. The suitable number of coefficients was determined by comparing the original and reconstructed banana shape， and then the principal component analysis was used to reduce the number of non-negative coefficients. Finally， the classification was achieved by the fuzzy C-means clustering. The results demonstrated that this machine method could be applied for banana shape identification in practice. This study provided a technological means for the value increase of banana and its secondary products， and also might be the basis of realizing the shape recognition of banana hand and bunch in the future.

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Advance of the research on molecular mechanism underlying yield traits in rice

2014, 26(1): 0-259.

Abstract ( 374 )

With a large number of genes contributed to high\|yield in rice had been mapped and cloned， the molecular mechanism underlying yield traits has been gradually understood in rice. IPA1 gene and four other genes directly controlling tiller development were cooperated with each other for conferring the plant architecture traits. Up to now， it was reported that 11 genes participated in the panicle formation， 3 of which were involved in the basic biological processes in panicle development， while 6 genes regulated the rate of spikelet formation， and 2 genes regulated the duration of panicle differentiation. Genes for determination of grain size or grain weight had five members， which were involved in grain length， grain width and the filling of grain, respectively. Cloning and understanding the molecular nature of genes controlling rice yield traits may be a fundamental implication for rice genetic improvements.

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