Acta Agriculturae Zhejiangensis

Measurement uncertainty in genetically modified maize DAS-4-40278-9 estimated by Monte Carlo method

SONG Jun, YE Xianlin, YIN Quan, ZHANG Fuli, WANG Dong, LI Jie

2018, 30(5): 681-687. DOI: 10.3969/j.issn.1004-1524.2018.05.01

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The GUM method is a commonly used way for evaluating measurement uncertainty (MU) in the field of products detection,whereas the approach has some limitations such as the hypothesis of normal distribution in output quantity. To provide a better and more suitable method for the estimating MU of genetically modified organism (GMO) detection,the Monte Carlo method (MCM) was employed to evaluate the MU of GM corn DAS-4-40278-9 in mixed sample. The results showed that the relative content of the GM corn DAS-4-40278-9 (C) was 1% with a small MU of 3.07×10^-4;the C value fell in a 95% coverage interval of 0.94%-1.06% indicating a high detection quality. In addition,the probability density distribution of the C was presented in a normal distribution,which was consistent with the assumption mentioned above in GUM method. Because MCM can avoid the limitation in traditional GUM method,MCM should be adopted in the evaluation of MU of GMO detection.

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Preliminary study on resistance identification and mechanism of maize inbred lines to Asian corn borer

WU Degong, FANG Wenhao, CHEN Huan, DU Junli, YU Haibing, WANG Changjin

2018, 30(5): 688-694. DOI: 10.3969/j.issn.1004-1524.2018.05.02

Abstract ( 426 )

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Asian corn borer, Ostrinia furnacalis, is an important maize pest. The most economical and effective measurement to control it was to breed resistant varieties. In 2016, the resistance level of 1 463 common maize inbred lines to maize borer was evaluated in Fengyang, Anhui and Ledong, Hainan. At the same time, the physicochemical characters, such as plant height, ear height, leaf number, length, width and angle of spike three leaves, stem diameter, leaf thickness, leaf hardness, tannin content, wax content, soluble sugar and soluble protein content, were measured. The relationship between borer resistance and physicochemical characters was analyzed. The identification results showed as follows: there were 142 highly-resistant inbred lines, 317 resistant inbred lines and 322 mid-resistant inbred lines and zoophobocus inbred lines accounted for 53.38%. Correlation analysis showed that the plant height, ear height, leaf number, leaf length, leaf width, leaf angle, stem diameter, and wax content of maize inbred lines were not significantly related to borer resistance. However, the leaf thickness, leaf hardness, tannin content, soluble sugar and soluble protein content were significantly correlated with maize resistance.

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Cadmium uptake characteristics of different rice cultivars

MAO Yaxi, FU Jianrong, MA Junwei, ZOU Ping, LEI Tinghai, LI Huan, LI Yuanwei, HUANG Yixiao, WANG Lingli

2018, 30(5): 695-701. DOI: 10.3969/j.issn.1004-1524.2018.05.03

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The cadmium (Cd) absorption and accumulation characteristics of four rice varieties, i.e. Yongyou 538, Chunyou 84, Jiahe 218 and Xiushui 134 were studied by Cd stress experiment in paddy fields in Shaoxing, Ninghai and Wenling, Zhejiang Province. It was shown that four rice varieties possessed Cd low accumulation characteristics in rice grain. When Cd concentration in rice grain in Yongyou 538, Chunyou 84, Jiahe 218 and Xiushui 134 was above 0.2 mg·kg^-1, the corresponding soil Cd concentrations in treatments were >1.41, 1.41, 0.91, 0.52 mg·kg^-1, respectively. The linear regression equation could be used to fit the relationship between Cd content in rice grain and the total Cd concentration in paddy soils. Under rice safe production, the order of the threshold value of the Cd concentration in paddy soil of four rice varieties was as Yongyou 538>Chunyou 84>Jiahe 218>Xiushui 134, and the order of the threshold value of the Cd concentration in paddy soil of different soil types was as clay degleyic paddy soil>tidal mud percolating paddy soil>tital mud redoximorphic paddy soil. In this study, the critical value of soil Cd concentration under rice safe production was higher than that of the current national soil environmental quality standard (GB 15618—1995).

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Expression and subcellular localization of bone marrow stromal antigen 2 gene of Capra hircus

ZHANG Li, LIU Teng, MIAO Qiuhong, TANG Jingyu, ZHU Jie, DONG Dandan, CHEN Zongyan, WANG Guijun, LIU Guangqing

2018, 30(5): 702-706. DOI: 10.3969/j.issn.1004-1524.2018.05.04

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BST-2 gene (Capra hircus bone marrow stromal antigen 2 gene) was amplified and cloned into pGEX-4T-1 and p3×Flag-CMV-14, respectively. The recombinant plasmids were named as pGEX-BST-2 and p3×Flag-BST-2, respectively. Firstly, pGEX-BST-2 was transformed into E.coli BL21(DE3), then the protein expression was induced by IPTG. The results of SDS-PAGE and Western blot analysis showed that the molecular weight of the recombinant protein was about 42 ku. Finally, Vero cells were transfected with p3×Flag-BST-2 through Lipofectamine 2 000 reagent, and the cells were analyzed by confocal microscopy.

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Expression analysis of crest phenotype related genes from crested white duck

ZHANG Yang, YAO Wencheng, WANG Bin, LU Lizhi, WANG Zhaoshan, XU Qi, CHANG Guobin, CHEN Guohong

2018, 30(5): 707-710. DOI: 10.3969/j.issn.1004-1524.2018.04.05

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To study the molecular mechanism of crest phenotype from crested white duck, crested white ducks and Liancheng white ducks were selected to detect the differential expressions of Hoxc8, EphB2, EphA2, EphA3 genes by real time quantitative PCR. Tissue distribution profiles were constructed and the results may help to understand the molecular mechanism of crest phenotype.

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Expression and distribution of Mel 1b mRNA and protein in various tissues of duck

LIU Wenju, WANG Shujuan, LIU Xiaoli, PANG Xunsheng, WANG Like

2018, 30(5): 711-716. DOI: 10.3969/j.issn.1004-1524.2018.05.06

Abstract ( 543 )

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A broad physiological function of melatonin is mediated by binding to melatonin receptors (Mel 1a, Mel 1b and Mel 1c). Melatonin receptors are widely distributed in the body, but vary according to subtypes, species and tissues. The study investigated the expression, distribution of Mel 1b in different tissues of ducks (including heart, spleen, brain, kidney, liver, lung, pancreas and breast muscle) using RT-PCR, real time PCR and immunohistochemistry. The results showed that Mel 1b mRNA was extensively expressed in heart, spleen, brain, kidney, liver, lung, pancreas and breast muscle of ducks, and Mel 1b protein was distributed in brain, lung, liver, skeletal muscle, kidney, heart and pancreas. The expression levels of Mel 1b mRNA were different among various tissues. The highest expression of Mel 1b mRNA was found in ovary, followed by the lung. Compared with the expression level of Mel 1b in brain, the expression levels in spleen, heart kidney and liver were higher, but those in breast muscle and pancreas were lower. Thus, Mel 1b was widely distributed in duck tissues, which further confirmed its involvement in the broad physiological functions of melatonin, especially the regulation of reproduction.

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Effect of plasmid-mediated tet(A) variant on tigecycline resistance in Klebsiella pneumoniae

JING Wei, CHENG Yumeng, SHAN Xinxin, LI Dexi, YAO Hong, DU Xiangdang

2018, 30(5): 717-721. DOI: 10.3969/j.issn.1004-1524.2018.05.07

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Tigecycline is one of the last choices to treat multidrug-resistant Klebsiella pneumoniae infection. However, the emergence of the tigecycline-resistant Klebsiella pneumoniae has posed a serious threat in clinical treatment. A clinical tigecycline-resistant Klebsiella pneumoniae strain was isolated, and a tigecycline-resistant plasmid was obtained by electroporation. Functional analysis revealed that two frameshift mutations occurred in the tet(A) gene on this plasmid. The antimicobial susceptibility tests indicated that the mutations could increase the minimal inhibition concentrations of tigecycline by 8 folds, whlie the MIC values of minocycline and tetracycline could increase 128 times due to these point mutations. The study confirmed that the plasmid-mediated tet(A) variant could confer the resistance to tigcycline in Klebsiella pneumoniae, which not only broaded the theoretical understanding about the resistance mechanism of Klebsiella pneumoniae to tigecycline, but also provided a basis to control the spread of such resistant gene among the gram-negative organisms.

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Effects of compound traditional Chinese medicine preparation on thymus tissue structure and caspase-3 protein distribution of SD rats

ZHOU Jinxing, QIAN Zhaoquan, JIN Erhui, LIU Wenju, JIN Guangming

2018, 30(5): 722-729. DOI: 10.3969/j.issn.1004-1524.2018.05.08

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To study the effects of compound traditional Chinese medicine preparation on the thymus tissue structure and Caspase-3 protein distribution of SD rats, which consist of Atractylodes macrocephala, Crataegus pinnatifida Bunge, Astragalus membranaceus (Fisch.) Bunge, Isatis tinctoria, Poria cocos Wolf, Pericarpium citri Reticulatae, Radix glycyrrhizae and Dioscoreae rhizoma. 60 SD rats (SPF animal), were randomly divided into control treatment group, compound traditional Chinese medicine soup 1.5, 3.0 and 6.0 mL·kg^-1 treatment groups, with 15 rats in each. After pre-feeding for one week, 3.0 mL distilled water, 1.5 mL, 3.0 mL and 6.0 mL soup according to mass per kilogram were applied by timing gavage, respectively. On the 90^th day, each treatment group rats were fasted and banned water for 3 h, then weighed, anaesthetized and put to death. The full thymus were taken and weighed, and were immediately immersed into the 4% polyformaldehyde phosphate buffer and fixed for 72 h, for making paraffin section, by HE, reticular fiber and caspase-3 immunohistochemical dyeing, microscopic observation, Image-pro PLUS 6.0 software analysis and photography. The results showed that 3.0 mL compound traditional Chinese medicine preparation treatment could increase obviously cortex of thymus according to the mass per kilogram, and the growth index of thymus gland was significantly higher than that of the control group (P<0.05). The reticular fibers were mainly distributed in the thymus medulla of SD rats, and the content of reticular fiber in 3.0 mL·kg^-1 dose group was significantly lower than that of the control group. Each test group could inhibit the appearance of caspase-3 protein in different degrees, the inhibition effect of the 3 mL·kg^-1 dose group was the most significant. It indicated that 3 mL·kg^-1 compound Chinese medicine preparation treatment could improve the thymus tissue structure of SD rats, delay its structure and function degradation.

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Isolation and identification of pathogenic Aeromonas veronii from Misgurnus anguillicaudatus

YUAN Xuemei, YAO Jiayun, LIN Lingyun, PAN Xiaoyi, XU Yang, YIN Wenlin, SHEN Jinyu

2018, 30(5): 730-737. DOI: 10.3969/j.issn.1004-1524.2018.05.09

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The present study was conducted to clarify species and drug resistance of pathogen from the diseased Misgurnus anguillicaudatus in a farm. Pathogenic bacteria from the liver of the diseased M. anguillicaudatus were examined using conventional methods and isolated. The pathogenic bacteria named strain NQ150728 were isolated and identified as Aeromonas veronii by means of VITEK-2 identification system and 16S rRNA and gyrB gene sequence analysis. The results of artificial infection test showed that the isolate could induce symptom of natural disease and caused death in 5 days. In addition, strain NQ150728 was highly sensitive to 24 kinds of drug such as ceftriaxone, riftriaxone, enrofloxacin and tetracycline.

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Study of anti-inflammatory effects of extracts from Zornia diphylla (L.) Pers in mice

CHEN Xiuying, ZHANG Yuchen, XU Xiangkun, FENG Xuejian

2018, 30(5): 738-743. DOI: 10.3969/j.issn.1004-1524.2018.05.10

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The peritoneal capillary permeability test, xylene induced auricular swelling test and carrageenan induced paw edema test were used to study the anti-inflammatory effect of Zornia diphylla (L.) Pers. Compared with the model control group, Zornia diphylla (L.) Pers inhibited the penetration of capillary vessel, decreased the swelling rate of auricle and paw in mice and relieved the pathological changes of the toes swelling and the exudation of PGE₂ in the inflammatory site after intragastric administration for 5 days. These results showed that Zornia diphylla (L.) Pers had inhibitory effects on acute inflammation in mice, and the mechanism may be related to the reduction of PGE₂ in the inflammatory sites.

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Study on toxicity of three kinds of triazole fungicides on zebrafish (Danio rerio)

GE Jing, JIANG Jinhua, CAI Leiming

2018, 30(5): 744-755. DOI: 10.3969/j.issn.1004-1524.2018.05.11

Abstract ( 545 )

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The acute toxicity of three kinds of triazole fungicides (tebuconazole, propiconazole, difenoconazole) to zebrafish was explored, and the joint toxicity of two drugs under conditions of equivalent toxicity and equivalent concentration was explored. At the same time, a six-day embryonic exposure was also carried out to investigate the developmental effect of three triazole fungicides. The study showed that the acute toxicity of the three kinds of triazole fungicides (tebuconazole, propiconazole, difenoconazole) to embryo, larvae and adult fish of zebrafish were medium; difenoconazole and propiconazole had synergistic effects on the embryo and fish larvae of zebrafish under conditions of equivalent toxicity and equivalent concentration; the rest combination of two drugs have antagonistic effects on the embryo and larvae of zebrafish. The results of developmental toxicity test indicated that the three triazole fungicides were more toxic to zebrafish at an early life stage and could induce a series of adverse symptoms in zebrafish embryos and larvae, including decreased hatching rate, abnormal heart rate and growth inhibition, etc.

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Combined effects of chlorpyrifos and chlorothalonil on early development stage of zebrafish

LI Jian, YANG Guiling, ZHAO Huiyu, WANG Qiang, ZHU Chaohua

2018, 30(5): 756-763. DOI: 10.3969/j.issn.1004-1524.2018.05.12

Abstract ( 458 )

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To evaluate the toxicity of chlorpyrifos and chlorothalonil in single dose and combinations, zebrafish was taken as biological subjects. By using the static method and real-time fluorescent quantitative PCR, the acute toxicity of chlorpyrifos or/and chlorothalonil on zebrafish embryos and their influence on the related genes of zebrafish larvae gonadal axis were studied. It was shown that chlorpyrifos and chlorothalonil were moderately and highly toxic to zebrafish embryos, respectively, as the 96 h-LC₅₀ (lethal concentration 50) was 2.139 mg·L^-1 and 0.353 mg·L^-1, respectively. The toxicivy of chlorpyrifos and chlorothalonil were synergistic to zebrafish embryos. In the joint exposure of chlorpyrifos and chlorothalonil, the 96 h-LC₅₀ was 0.38 mg·L^-1 and 0.063 mg·L^-1, respectively. Moderate concentration (26.7 μg·L^-1) and high concentration (106.7 μg·L^-1) of chlorpyrifos led to upregulation of the gene expression of the aromatase (CYP19α), vitellogenin (VTG1, VTG2), and estrogen receptor (ERα, ERβ1 and ERβ2). Low concentration of 1.1 μg·L^-1 chlorothalonil led to the upregulation of gene expression of VTG1, VTG2, ERβ1 and ERβ2. Combination of 6.7 μg·L^-1 chlorpyrifos+1.1 μg·L^-1 chlorothalonil led to the downregulation of gene expression of VTG1, ERβ1, ERβ2 and ERα in zebrafish compared with the control. The results indicated that chlorpyrifos and chlorothalonil had an estrogenic effect on zebrafish, while the combination reduced the effect of estrogenic interference. Therefore, the effects of combined pollution should be fully considered for harm assessment of pesticides to aquatic organisms.

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Study on fruit development and quality formation of (Myrica rubra) cv. Zaoxian

ZHU Tingting, LIANG Senmiao, ZHANG Shuwen, ZHENG Xiliang, REN Haiying, QI Xingjiang

2018, 30(5): 764-770. DOI: 10.3969/j.issn.1004-1524.2018.05.13

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In order to study the breeding potential of Zaoxian and the formation law of fruit development and quality, the dynamic changes of single fruit weight, vertical and transverse diameter, chromatic aberration, saccharide, acid, Vc, total phenol and flavonoid during the fruit development of Zaoxian and Zaose were determined. The results showed that the single fruit weight of Zaoxian increased rapidly from the white stage. The growth rate of fruit was obviously higher than that of Zaose, which was the performance of the early ripening of the fruit. The dynamic changes of the growth, chromaticity parameters and fruit quality of the two Myrica rubra fruits were basically the same but each was different. The change of color difference parameter can be used to analyze the color and maturity of Myrica rubra fruits. During the process of fruits ripening, the contents of soluble sugar increased gradually but the contents of oxalic acid, Vc, phenols and flavonoids decreased. The titratable acid and citric acid content increased firstly and then decreased. But the content of malic acid was firstly decreased but increased at anaphase in the growing periods of Myrica rubra fruits. This study confirmed that the fruit of Zaoxian matured earlier and it had higher soluble sugars, Vc and total phenols contents and lower titratable acid content.

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Cloning and prokaryotic expression of 1-deoxy-D-xylulose-5-phosphate synthase encoding gene (BaDXS1) in Brassica alboglabra

XUE Shengling, JIANG Min, CHANG Jiaqi, LIU Yang, WEI Lin, ZHOU Jiankun, ZHANG Fen, SUN Bo

2018, 30(5): 771-777. DOI: 10.3969/j.issn.1004-1524.2018.05.14

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The 1-deoxy-D-xylulose-5-phosphate synthase (DXS) encoding gene BaDXS1 was isolated from Chinese kale (Brassica alboglabra) leaves by using homologous cloning method. BaDXS1 contained an open reading frame of 2 139 bp in length, encoding 712 amino acids. Physical and chemical properties analysis indicated that the molecular weight of BaDXS1 protein is 77.21 ku, and the isoelectric point pI is 8.59. There was no transmembrane region, and BaDXS1 is located in chloroplast of plant cell. Amino acid sequences comparison results showed that the identities of BaDXS1 and those in Brassica oleracea, Arabidopsis thaliana, tobacco, tomato were all over 79%. Phylogenetic tree analysis showed that it had close relation with BoDXS. The prokaryotic expression vector pEASY-Blunt E1-BaDXS1 was constructed, and the protein was successfully expressed in the form of inclusion bodies in Escherichia coli.

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Establishment of complex growth-promoting rhizobacteria for watermelon and promoting effect on watermelon roots

XU Weihui, LYU Zhihang, SHI Yiran, JIANG Jiaying, HU Yunlong, WANG Zhigang

2018, 30(5): 778-786. DOI: 10.3969/j.issn.1004-1524.2018.05.15

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Plant growth-promoting rhizobacteria (PGPR) are important species resources of microbial fertilizer, and they play important roles in agriculture production. In this study, the capability of hormone-secreting and antagonistic effect among experimental strains were evaluated, combinations of complex bacteria were constructed, and effect of complex bacteria on watermelon roots were studied. The results showed that the all twelve bacteria strains could secrete indoleacetic acid (IAA), the best one was Streptomyces CL05, whose yield was 117.30 mg·L^-1. Sphingomonas CL03, Lysobacter CL02 and Microbacterium LS01 could not product siderophores, Microbacterium LS01, Microbacterium HYL03 and Pantoea ananatis HYL01 could not secrete gibberellin. Furthermore, three groups of complex bacteria were generated based on the antagonistic experiments, which were named as R1, R2 and R3, respectively. In unsterilized soil, complex bacteria R2 significantly increased root length, root surface area, root volume, root number and root dry weight of watermelon seedlings by 79.0%, 46.8%, 57.1%, 162.7% and 49.1% compared with the control, respectively. In sterilized soil, complex bacteria R2_S increased root length, root surface area, root volume, root number and root dry weight of watermelon seedlings by 191.8%, 302.4%, 160.0%, 206.5% and 139.6% compared with the control, respectively, and decreased root mean diameter of watermelon seedlings by 17.5%. Interestingly, the promoting effect of R2 on watermelon roots was better in sterilized soil than in unsterilized soil. Overall, the study indicated that complex bacteria R2 could significantly promote the growth of watermelon roots, and the ability of gibberellins secretion in R2 (CL03, LS05, HYL02, WZW03) exceeded that in R1 and R3, which may be one of promoting mechanisms of complex bacteria R2.

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Proteomic analysis of black plum leaf treated with immunity inducer

ZHANG Qing, XIAO Wenfei, QIU Jieren, CHEN Chuwei, XIN Ya, CHAI Weiguo, RUAN Songlin

2018, 30(5): 787-796. DOI: 10.3969/j.issn.1004-1524.2018.05.16

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The plum variety Friar plum was used as test material. Black plum leaves were treated with immunity inducer Baokangling No.1. Proteomic analysis and determination of physiological and biochemical indexes of treated leaves were carried out. The results showed that area, thickness, chlorophyll content and PPO activities of black plum leaf treated with immunity inducer Baokangling No.1 all increased. The cross section of blade showed that the palisade tissue, spongy tissue and lower epidermis were obviously thickened after treatment with immunity inducer, which was probably responsible for leaf thickening and increased chlorophyll content. Proteomic analysis showed that a total of 157 differentially expressed proteins were screened, including 75 up-regulated proteins (up to 1.5 fold and P<0.05) and 82 down-regulated proteins (down 1.5 fold and P<0.05).GO classification and KEGG pathway analysis showed that differentially up-regulated protein (or genes) were enriched and assigned to the chloroplast, chloroplast membrane and chloroplast matrix function, whereas differentially down regulated proteins (or genes) were enriched and assigned to chloroplast matrix, the chloroplast membrane and photosystem Ⅰ. Both of them were involved in photosynthesis. Taken together, the immunity inducer Baokangling No.1 may promote the growth of black plum leaves via regulating the expression of proteins related to photosynthesis.

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Differential biological characteristics between closely related tea geometrid species, Ectropis obliqua and Ectropis grisescens

BAI Jiahe, TANG Meijun, YIN Kunshan, WANG Zhibo, XIAO Qiang

2018, 30(5): 797-803. DOI: 10.3969/j.issn.1004-1524.2018.05.17

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The sibling pests, Ectropis grisescens and Ectropis obliqua, are morphologically similar. However, the potential differences between their biological characteristics especially the body-length, weight, developmental duration and population growth rates of each development stage remains unclear. In the current study, we characterized and compared this issue based on laboratory observation. Firstly, the body-length and weight of the old larvae, pupae and moths were significantly different, with higher levels observed on E. grisescens in comparison to those of E. obliqua. Secondly, the periods of one generation between these two pest species were distinct. E. grisescens needed 36.6 d to complete one generation, whereas E. obliqua required at least 39.7 d. Finally, the generational survival rate and population growth rate in E. obliqua was 0.28 and 24.9, respectively, which was approximately half of that in E. grisescens. Hence, we proposed that E. grisescens had larger body size, faster developmental rate and stronger reproductive capacity than E. obliqua.

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Expression of antimicrobial peptides thanatin tandem fusion in Escherichia coli

LIU Wei, LI Qinghai, LIU Renhu

2018, 30(5): 804-809. DOI: 10.3969/j.issn.1004-1524.2018.05.18

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The antibacterial peptide thanatin is a broad spectrum cationic antibacterial peptide, which can significantly kill pathogenic fungi. In order to improve the expression level of thanatin, three copies of thanatin monomer and MBP protein was co-expressed in Escherichia coli. The expression products were identified by Western blot and mass spectrometry after enzyme digestion and purification. The results showed that the three copies of thanatin fusion protein, 3×thanatin, were successfully expressed in E. coli and had good inhibition activity on Sclerotinia sclerotiorum. The minimum concentration of inhibition on S. sclerotiorum was 6.6 μmol·L^-1 for 3×thanatin, which was about ten times higher than 1×thanatin.

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Combined effects of salinization and acidification on cadmium activation in greenhouse soil

WANG Jingwen, XIE Guoxiong, LI Dan, ZHANG Mingkui

2018, 30(5): 810-816. DOI: 10.3969/j.issn.1004-1524.2018.05.19

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Soil salinization and acidification are common in greenhouse soil. Both of them could change the balance of chemical elements in soil, and thus affect the chemical behavior of heavy metals in soil. To understand the effects of simultaneous salinization and acidification on Cd activity in greenhouse soil, changes of soil water-soluble Cd in soil and Cd uptake by vegetable as functions of soil pH, salinity and salt accumulation were observed using simulation experiments. It was shown that water-soluble Cd content in soil and Cd accumulation in upper parts of vegetable increased obviously with the increase of soil acidification and salinization. The change of soil water-soluble Cd concentration in soil with increasing salinity was most obvious at pH 5.00-5.50. For soil with total Cd of 0.55 mg·kg^-1 and soil salinity of 2.50 g·kg^-1, the concentrations of water-soluble Cd in soil increased by 10.53%, 44.00% and 51.06% , respectively, under pH of 6.15, 5.50, and 5.00, as compared with the control soil without salinization. Correspondingly, Cd accumulation in upper parts of vegetable increased by 20.00%, 16.85% and 36.36%, respectively. Meanwhile, concentration of water-soluble Cd in soil and Cd content in the upper parts of vegetable increased by about 4 and 8 times of the control, respectively, when soil pH was reduced from 6.15 to 4.00. The activation of Cd in soil varied with ion composition of salt, and the effect decreased in the order of Ca²⁺,Mg²⁺>K⁺>NH4+ for cation, and Cl^->SO42->NO3- for anion. The results suggested that acidification and salinization occurred simultaneously in the protected soil could significantly increase the activity of soil cadmium. Those two processes had a superimposed effect on the activity of soil Cd.

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Soil microbial community structure, labile organic carbon and nitrogen and enzyme activities in paddy field and upland affected by long-term fertilization systems

WANG Guiyue, SU Ting, HAN Hailiang, TAN Heping, BAO Fei, ZHAO Fucheng

2018, 30(5): 817-824. DOI: 10.3969/j.issn.1004-1524.2018.05.20

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To investigate the effects of long-term fertilization systems on soil microbial community structure, labile organic carbon and nitrogen, and enzyme activities in yellow sand paddy field and upland, a field experiment was conducted at the experimental station of Dongyang Institute of Maize Research, Zhejiang, China in 2009. The experiment consisted of six treatments with three replicates and was arranged in a completely randomized design: PCK, no fertilization in paddy field; PCF, conventional fertilization in paddy field; PSTF, formulated fertilization by soil testing in paddy field; PSTF+OF, formulated fertilization by soil testing with organic manure in paddy field; DCF, conventional fertilization on upland; DSTF+OF, formulated fertilization by soil testing with organic manure on upland. Soil nutrients, enzyme activities, microbial biomass and community structure were determined in 2015. It was shown that compared with no fertilization in paddy field (PCK), all fertilization treatments significantly (P<0.05) increased soil available phosphorus and potassium contents, and decreased soil pH value. All treatments had no significant effects on quantity of soil culturable bacteria and actinomycetes, yet PSTF+OF significantly (P<0.05) increased the quantity of soil culturable fungi. Formulated fertilization by soil testing with organic manure (PSTF+OF and DSTF+OF) significantly (P<0.05) increased soil microbial carbon and nitrogen contents both in paddy field and upland. All treatments had no significant effect on soil cellulose activity, while formulated fertilization by soil testing with organic manure (PSTF+OF and DSTF+OF) significantly (P<0.05) increased soil dehydrogenase and catalase activities. In general, application of chemical fertilizers with organic manure could improve soil fertility.

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Effects of no tillage and stubble mulch on soil meso-microfauna communities

MA Nan, QIAN Ruixue, YANG Huimin, CHEN Zhiwen, JIANG Yunfeng

2018, 30(5): 825-831. DOI: 10.3969/j.issn.1004-1524.2018.05.21

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In order to evaluate the effects of no-tillage and stubble mulch on the soil meso-microfauna assemblages, the abundance, community composition and seasonal variation of soil meso-microfauna were investigated in the Conservation Tillage Research and Development Station of Chinese Academy of Sciences in Lishu County, Jilin Province in April, July and October, 2015. A total of 18 309 soil meso-microfauna were obtained, which belonged to 2 phyla, 6 classes, 16 orders. Among them, the dominant group is Oribatida, Mesostigmata, Hypogastruridae and Isotomidae. The individual density of soil meso-microfauna in no-tillage plots was higher than that in other treatments, while the highest taxon richness and diversity index was appeared in the stubble mulch plot. The highest individual density was recorded in October for all treatment plots. Major soil animal groups differed in response to tillage methods and seasonal variations. Compared with conventional tillage, no-tillage and stubble mulch could increase the individual density of major soil animal groups.

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Characteristics of soil basic respiration in different altitude forests in Sejila Mountain, southeast of Tibet

YANG Hong, LIU Heman, CAO Lihua, XU Changchang, CAO Jianting, SAI Man

2018, 30(5): 832-839. DOI: 10.3969/j.issn.1004-1524.2018.05.22

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In order to clarify the responses of soil basal respiration to temperature changes at different altitudes, and to provide scientific basis for predicting soil carbon dynamics at different altitudes in the future climate change scenarios, the forest soils at different altitudes in Sejila Mountain were studied by laboratory simulated heating experiment. It was shown that soil basal respiration rate and soil accumulated carbon flux increased with the increase of temperature. With the deepening of the soil layer, soil basal respiration rate was decreased. Soil respiration rate of all layers and altitudes increased at first and then decreased with the extension of simulated culture time, while local parts showed a characteristic of concussion change. There was a significant (P<0.01) negative correlation between soil basal respiration rate and culture time, which could be described as exponential function. The cumulative carbon flux of soil basal respiration showed an increasing trend with the prolongation of culture time, and the increases of soil accumulated carbon flux were obvious in the first 14 days, and then gradually became stable. There was a significant (P<0.01) positive correlation between accumulated soil carbon flux and incubation time, which could be described as logarithmic function. In summary, elevated temperature would accelerate carbon emission via surface soil respiration of forest ecosystem.

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New requirements for grapes of national food safety standard GB 2763-2016

LI Zhen, SUN Caixia, DAI Fen, ZHENG Weiran, YU Guoguang, YAO Jiarong, WANG Qiang

2018, 30(5): 840-847. DOI: 10.3969/j.issn.1004-1524.2018.05.23

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National food safety standard ‘Maximum residue limits for pesticides in food' (GB 2763-2016) puts forward more new requirements for pesticide residues in food, maximum residue limit and detection methods. In this paper, the changes in the provisions of grapes or berries in the new standard were summarized, and were compared with relevant international standards, to provide reference for the development of related industries.

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Extraction technology of lecithin from follicle

YE Mengdi, TANG Honggang, XIAO Chaogeng, YANG Huijuan, CHEN Di, CHEN Lihong

2018, 30(5): 848-853. DOI: 10.3969/j.issn.1004-1524.2018.05.24

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In order to improve the utilization rate and additional value of the by-products of layer, the study explored the optimum condition of extraction through single factor and orthogonal test with follicle as material. Results showed that the optimum extraction conditions were obtained as follows: ethanol concentration 95%,extraction temperature 25 ℃,extraction time 25 min and solid-liquid ratio 1∶12.Under these conditions, the yield and extraction rate of lecithin were 45.35% and 90.14%, respectively.

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Application of foodborne lactic acid bacteria in improving tobacco quality

YANG Juan, MI Qili, XIONG Wen, ZHU Ruizhi, LOU Mumeng, YAO Jianhua, LUO Yiyong

2018, 30(5): 854-862. DOI: 10.3969/j.issn.1004-1524.2018.05.25

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In order to explore the possibility of lactic acid bacteria (LAB) for improving the smoking quality of tobacco, two strains of LAB (Lactobacillus plantarum L8 and L14) were isolated and identified from Douchi, a Chinese traditional fermented food. Using L8 and L14 for cut tobacco fermentation, sensory evaluation results indicated that the smoke of fermented cigarette was fine and smooth, with a reducing miscellaneous gas and offensive irritancy, an obvious acid fragrance, and a clean aftertaste. On the basis of sensory evaluation, the fermentation condition of strain was optimized by orthogonal experiment. The results showed that the optimal aroma-producing condition was as follows: the inoculum size of 1×10⁸ CFU·mL^-1, the fermentation temperature of 37 ℃, the bacteria age of 10 h, and the tobacco producing area of Baoshan, Yunnan. Using the cut tobacco of commercial cigarette (Yunyan Ziyun) as fermented material, the results of sensory evaluation proved again that L8 and L14 could improve significantly the smoking quality of tobacco. The main chemical compositions in the fermented cut tobacco became more harmonious. The contents of total acid (especially of acetic acid) and total aldehyde (particularly of 3-methyl butyral) were significantly increased and reduced, respectively. The above results showed that LAB have great potential in improving tobacco quality.

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Analysis of dynamic change characteristics of iron ore and its surrounding vegetation based on multi temporal remote sensing

PANG Dong, TAN Lin, HE Bingyu

2018, 30(5): 863-871. DOI: 10.3969/j.issn.1004-1524.2018.05.26

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In Xinjiang iron ore and its surrounding area,using 2007—2016 multi temporal TM/ETM+and OLI images,analysis of the iron ore and its surrounding large areas of vegetation cover index (FVC) and the growth of vegetation condition index (VCI), the vegetation coverage, temporal and spatial changes of vegetation growth and the effect of iron ore mining on the surrounding environment were evaluated, to provide decision support for mine environment. The results showed that FVC and VCI index can reflect the low level of coverage vegetation and vegetation growth status in the study area, and there was almost no actual vegetation growth in iron ore area, which could provide theoretical support for ecological risk prevention. In 2007—2016, the overall vegetation coverage and vegetation growth in the study area fluctuated upward, and the lower vegetation coverage degree and poor vegetation growth status degree developed to a better level at the same time. The higher grades of FVC and VCI were mainly distributed in the southern and southwestern slope. But the iron ore mining area increased year by year, and the area and the surrounding area has caused serious degradation of vegetation and ecological damage, which could easily lead to soil erosion and landslides and other disasters. Thus, prevention and control measures should be taken in time.

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Spatial-temporal distributions of pork price and its influencing factors analysis

GU Jingqiu, PENG Cheng, WU Huarui, WU Jianzhai

2018, 30(5): 872-879. DOI: 10.3969/j.issn.1004-1524.2018.05.27

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In this paper, pork prices in 30 provinces from 2005 to 2014 are analyzed for exploring spatial and temporal distribution pattern and the coupling relationship between pork price and 11 influencing factors of supply, demand and cost. The results showed that pork prices showed a significant positive correlation between provinces from 2005 to 2008. After 2008, the correlation decreased and there was a significant difference in spatial distribution. In terms of influencing factors, the pork price was positively correlated with the price of pig, piglet and corn.

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