Acta Agriculturae Zhejiangensis

Preparation and application of multiclonal antibody against plant cysteine proteinase

PENG Qiqi, YANG Jian, LIAO Qiansheng, ZHANG Hengmu

2018, 30(6): 881-885. DOI: 10.3969/j.issn.1004-1524.2018.06.01

Abstract ( 748 )

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Cysteine proteases (CysPs) are a family of important proteases that are involved in a wide range of plant physiological processes. To characterize such proteinase, a gene encoding CysP was amplified by RT-PCR from Nicotiana benthamiana plants and ligated into pEASYTM-T5 Zero vector. The insertion was sequenced and then sub-cloned into the prokaryotic expression plasmid pGEX-6P1, named as pGEX-NbCysP. The plasmid was transformed into Escherichia coli BL21plysS for inducible expression. The recombinant NbCysP fusion protein was purified with affinity chromatography and used for producing polyclonal antibody by immunizing rabbits. In immunoblotting assays, the polyclonal antibody could react strongly with the recombinant NbCysP protein as the presence of a single band, indicating that the antibody was specific against the proteinase. These results laid a foundation for further characterization of its function.

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Effects of combined symbiotic nitrogen-fixing bacteria on growth of wheat seed under different environmental factors

DENG Tiantian, CHEN Xi, HOU Yumeng

2018, 30(6): 886-892. DOI: 10.3969/j.issn.1004-1524.2018.06.02

Abstract ( 882 )

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Three species of co-symbiotic nitrogen fixation bacteria were obtained from 13 strains of wheat. Different strains were determined by the single factor batch experiments to obtain the optimal culture time, temperature, pH, glucose concentration, NaCl concentration and nitrogen concentration; furthermore, to investigate the effects of the single strain and combination groups on the growth of wheat seeds. The results showed that under the conditions of pH 7, the temperature of 35 ℃, NaCl concentration of 5%, glucose concentration of 10%, nitrogen concentration of 3 g·L^-1, culture time of 36 h, the combination of group action showed better effect than the single strain, and compared with the control, the germination rate was increased by 4% on average, the germination potential, germination index also increased. To some extent, the combinatorial bacteria group can better adapt to the changeable environment, playing a stable role in promoting the yield.

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Proteomic analysis of maize seedling leaves in response to heat stress

SHI Jiang, ZHAO Lin, ZHU Yueqing, LOU Xuping, YU Jianzhong, RUAN Songlin, CHEN Wenyue

2018, 30(6): 893-908. DOI: 10.3969/j.issn.1004-1524.2018.06.03

Abstract ( 625 )

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Maize is a warm-tempered crop, but it is sensitive to high temperature weather during its growth and development, which is easy to cause the decline of yield. In this study, seedlings of a heat-resistant maize variety Xiantian No. 5 as experimental materials were treated for 0, 0.5, 3.0 h at 42 ℃, and leaf proteins from all treatments was extracted to carry out screening of differential proteins, and then the GO classification and KEGG pathway were analyzed. The results showed that 181 proteins including 95 up-regulated proteins (quantitative ratio>1.20 and P<0.05) and 86 down-regulated proteins (quantitative ratio<0.83 and P<0.05) were identified. Compared with the control, the number of up-regulated proteins increased with the increase of heat treatment time, while the number of down-regulated proteins decreased. GO classification indicated that in these differentially up-regulation proteins, there were 2 proteins enriched and designated to 2 cell components, 18 proteins enriched and designated to 8 molecular functions and 15 proteins enriched and involved in 17 biological processes. Similarly, in differentially down regulated proteins, there were 2 proteins enriched and distributed in 8 cell components, 29 proteins were enriched and designated to 11 molecular functions. 45 proteins involved in 18 biological processes. KEGG pathway analysis revealed that 16 differentially up-regulated proteins were enriched and designated to participate in 6 pathways, while 14 differentially down-regulated proteins were enriched and designated to participate in 8 pathways. Domain analysis revealed that 27 differentially up-regulated proteins were enriched and designated to 22 various domains, while 34 differentially down-regulated proteins were enriched and designated to 21 different domains. According to the searching information in NCBI database, we found that 5 proteins of Dna J molecular chaperone family including heat shock protein, thioredoxin H type, dehydrogenase and ubiquitin binding enzyme were likely to be closely related to heat resistance. In conclusion, identification of proteins related to heat stress response can provide an insight to further revealing the underlying molecular mechanism of maize response to heat stress.

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Cloning and expression analysis of StDWF1 in Solanum tuberosum

TANG Xiao, DENG Mengsheng, ZOU Xue, Li Liqin, ZHU Yuanzhi, WANG Xiyao

2018, 30(6): 909-917. DOI: 10.3969/j.issn.1004-1524.2018.06.04

Abstract ( 956 )

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The DWF1 is a key gene in brassinosteroid (BR) biosynthesis pathway. To explore the function of StDWF1 in the growth and development of potato, this experiment was based on the potato variety Favorita test-tube plantlets, and we cloned the full-length sequence of StDWF1 gene. The open reading frame (ORF) of StDWF1 gene was 1 704 bp, and encoded 567 amino acids. The molecular weight of protein was 66.08 ku, and the isoelectric point (pI) was 7.96. Agrobacterium tumefaciens-mediated transient expression was carried out in tobacco, and laser scanning confocal microscopy showed that potato StDWF1 was located in cytoplasm. Fluorescence quantitative analysis showed StDWF1 expression quantity was the highest in new leaves, followed by old leaves, stolon, roots, stems and tubers. We analyzed the expression of StDWF1 in potato leaves at the growth period, and the results showed that the highest expression quantity was in potato seedling stage. The results of this experiment laid the theoretical foundation for further interpreting the regulation mechanism of BR in the potato growth and development.

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Optimization of efficient regeneration system in stem of potato using response surface methodology

JIANG Linjuan, ZOU Xue, HUANG Xueli, NI Su, LI Liqin, YANG Shimin

2018, 30(6): 918-925. DOI: 10.3969/j.issn.1004-1524.2018.06.05

Abstract ( 755 )

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To establish the optimal hormone concentrations of culture medium for efficient regeneration of potato stem callus tissue, as well as build a new system for tissue culture and rapid propagation technology, stem of Chuanyu No.10 was employed as explants to induce callus tissue, and the Box-Behnken design method of response surface methodology (RSM) was used to research the effect of TDZ, 2,4-D and GA₃ on the differentiation of potato callus. Meanwhile, the hormone concentrations were optimized. The results showed that the sensitivity of the hormone concentrations in culture medium to potato callus budding from the response surface method optimization model is TDZ>GA₃>2,4-D. The determination conefficient R² of the model was 0.992 1, the optimal medium for adventitious bud differentiation from stem callus of Chuanyu No.10 was MS+2.02 mg·L^-1 TDZ+0.08 mg·L^-1 2,4-D+2.25 mg·L^-1 GA₃.In this condition, the budding rate was actual measured, which was consistent with the predicted values. That confirmed the validity of the model.

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Ability to break through blood-brain barrier of porcine Japanese encephalitis virus SCYA201021 strain after infection in mice

CHAI Chunxia, WANG Qiao, LAI Yalan, XU Yixuan, WEN Xintian, CAO Sanjie, HUANG Xiaobo, WEN Yiping, ZHAO Qin, WU Rui

2018, 30(6): 926-931. DOI: 10.3969/j.issn.1004-1524.2018.06.06

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In the present study, SPF Kuming mice were inoculated with SCYA201021 strain (F122 generation, attenuated strain of porcine Japanese encephalitis virus) by intracerebral injection in order to evaluate its potential value as the attenuated live vaccine, and its ability to break through the blood-brain barrier in mice. After infection in mice, pathological changes in mice brain were observed, and the E gene copes were measured by fluorescent quantitative PCR in the blood and brain at 6, 12, 18, 24 h after inoculation, respectively. The results indicated that the virulence of SCYA201021 strain (F122 generation) was significantly weakened as the half lethal does (LD₅₀) in suckling mice brain was only 4×10⁵ PFU·40 μL^-1. Moreover, the mice did not show pathological changes in the brain during the 24 h after intraperitoneal injection, and the quantitative PCR results were negative, indicating that the SCYA201021 strain could not break through the blood-brain barrier in mice and the SCYA201021 strain had high safety.

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Pathological study on duck-origin goose Parvovirus strain QH-L01 infection of Cherry Valley duck

CHEN Bing, XU Jing, LUO Qihui, HUANG Chao, LIU Wentao, WU Hong, CHEN Jingfei, CHEN Ping, GAO Qi, MA Wenjing, CHEN Zhengli

2018, 30(6): 932-938. DOI: 10.3969/j.issn.1004-1524.2018.06.07

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In the present study, the histopathological changes of various organs of Cherry Valley ducklings were explored under the condition of artificial infection, to clarify the pathogenesis and characteristics of duck-origin goose Parvovirus strain QH-L01. Thirty Cherry Valley ducks in experimental group were intramuscularly inoculated with 0.2 mL of 10^6.54 ELD₅₀ duck-origin goose Parvovirus strain, and the ducks in the control group were intramuscularly inoculated with 0.2 mL of sterilized normal saline per duck, to observe the pathological changes of various tissues/organs in each infective stage at 1, 3, 6, 9 d, respectively. On the 1^st day after infection, the lungs appeared punctate hemorrhage, partial lobular expansion, and intestinal tissue villous epithelial cells necrosed. On the 3^rd day after infection, brain, kidney, thymus, bursa fabricius showed pathological changes, and villus of duodenum and ileum slight necrosed, mucosa layer of cecum appeared slight necrosis. On the 6^th day after infection, lymphocytes in spleen increased, moderate necrosis was detected in villus of duodenum, ileum and mucosa layer of cecum. On the 9^th day after infection, the lesions were reduced or returned to normal. The mortality rate was 20% during the whole infection period.

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Evolutionary and molecular characteristics of M genes of porcine epidemic diarrhea viruses in western Fujian, China

DONG Bo, AN Yongshuai, DAI Ailing, LI Xiaohua, YANG Xiaoyan

2018, 30(6): 939-945. DOI: 10.3969/j.issn.1004-1524.2018.06.08

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To elucidate evolutionary characteristics of the memberane (M) genes of porcine epidemic diarrhea viruses in Fujian, China, 12 samples collected from Fujian province during 2015 to 2017 were screened by multiplex RT-PCR,M genes of PEDV viruses from positive samples were amplified by RT-PCR and cloned into pMD18-T vectors for sequencing respectively. The sequence data were done alignment and phylogenetic analysis with sequences from known reference strains. The M genes from 12 representative positive samples shared 100.0% identity at amino acid level. 12 representative positive samples are remote to classical strain CV777, SM98 and virulent DR13. 12 representative strains belong to the genotype variant strain. Molecular structure characteristics analysis showed that M gene is highly conservative and could be an optimal candidate protein for vaccine development.

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Mucous membrane organization structure characteristics and goblet cells distribution observation of Huanghuai white goat's intestinal tract

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

2018, 30(6): 946-952. DOI: 10.3969/j.issn.1004-1524.2018.06.09

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To observe mucous membrane organization structure characteristics and goblet cells distribution of Huanghuai white goat's intestinal tract, three adult healthy Huanghuai white goats were bleed from jugular vein to death, the samples of duodenum, jejunum, ileum, cecum, colon and rectum were taken immediately and immersed into 4% paraformaldehyde fixative to be fixed for 72 h. Tissue slices were thus made, and stained with HE and PAS. The results showed that epithelial cells of the duodenum and jejunum of Huanghuai white goat were highly columnar, and lymphocytes were distributed in lamina propria. The epithelial cells of the ileum mucosa were low columnar, and a large collection of lymph nodes were distributed in submucosa. The epithelial cells of cecum, colon and rectum were also low columnar, large intestinal gland were developed, and there were diffused lymphocytes between glands. The lymphoid nodule was seen in the lamina propria of rectum. The goblet cells in the epithelium secreted mucopolysaccharide in the form of apical secretion. The small intestinal goblet cells of Huanghuai white goat secreted neutral-mucopolysaccharide, while the large intestinal goblet cells of Huanghuai white goat secreted acid-mucopolysaccharide. The goblet cell's number of large intestine was significantly (P<0.01) higher than that of small intestine. The goblet cell number of colon were significantly (P<0.05) higher than those of the cecum and rectum, and the goblet cell number of ileum were significantly(P<0.05) higher than those of the duodenum and jejunum. Therefore, from duodenum to ileum, the epithelial cells gradually changed from high column to low column, the goblet cells were mainly distributed in the large intestine, and the highest number of goblet cells was found in the colon.

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Analyses of blood physiological and biochemical indices of Chinese Holstein cows during late period of perinatal in Sichuan rolling country

JIA Xianbo, YANG Shuhui, DING Peng, CHEN Shiyi, WANG Jie, HU Shenqiang, LAI Songjia

2018, 30(6): 953-960. DOI: 10.3969/j.issn.1004-1524.2018.06.10

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To establish the normal reference range for blood physiological and biochemical indices of Chinese Holstein cows in Sichuan rolling country, 46 blood indices, including 23 routine indices, 15 serum biochemical indices and 8 trace elements, were detected and statistically analyzed among 150 Chinese Holstein cows within the first three parities. The reference range of 46 blood physiological and biochemical indices were obtained for perinatal Chinese Holstein in Sichuan Province. 9 blood routine indices and 9 serum biochemical indices showed significantly (P<0.05 or P<0.01) differences among the three parities. The present study established the normal reference range for the 46 blood physiological and biochemical indices of Chinese Holstein cows in Sichuan Province, which would provide scientific references for the feeding, management, prevention and treatment of disease of China Holstein in Sichuan Province.

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Responses of Expansins and XTHs to different aluminum concentrationa in roots of tea plant [Camellia sinensis (L.) O. Kuntze]

NING Qiuyan, FAN Kai, WANG Min, SHI Yuanzhi

2018, 30(6): 961-969. DOI: 10.3969/j.issn.1004-1524.2018.06.11

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In order to investigate the effect of aluminium (Al) on root growth of tea plants [Camellia sinensis(L.) O. Kuntze], Anjibaicha tea plant was selected as study material with different Al concentrations for 28 d. Then, the phenotypic differences and parameters of roots, activities of the Expansins and xyloglucan endotransglucosylase (XET), and the gene expression level of Expansins and XTHs, which were related with root growth and key proteins of cell wall were assayed. The results showed that the Al could promote the root growth of tea plant on phenotypic and parameters. The treatment with 0.4, 1.0 mmol·L^-1 Al showed significant (P<0.05) difference with control, as the root total length was increased by 206% and 209%, respectively, the number of root tips was increased by 175% and 166%, respectively, the root surface area was increased by 227% and 286%, respectively, and the root volume was increased by 246% and 385%, respectively. Correspondingly, the gene expression level of CsEXPB14 and the expansins-like gene CsEXLA8 in the roots treated with 0.4 mol·L^-1 Al were significantly (P<0.05) higher than those without Al or with 4.0 mmol·L^-1 Al. The expansins activity in the roots with 0.4 mmol·L^-1 Al was higher than other treatments and was increased by 84.3% as compared to the control. The expression level of CsXTH14 in the roots with 0.4, 1.0 mmol·L^-1 Al were significantly (P<0.05) higher than the other treatments, and XET activity was increased with the elevated Al concentration, and was significantly (P<0.05) inhibited until the Al concentration reached up to 4.0 mmol·L^-1. The gene expression level of Expansins and XTHs, the activities of XET and Expansins showed a positive correlation with root growth of tea plant, which indicated that Al (≤1 mmol·L^-1) could increase XET and Expansins activity by promoting the gene expression of Expansins and XTHs, hence, promote root growth of tea plants.

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Effects of salt stress on anthocyanin content and activities of antioxidant enzymes in leaves of Photinia frasery

LIN Yicheng, FU Qinglin, GUO Bin, LIU Chen, DING Nengfei

2018, 30(6): 970-977. DOI: 10.3969/j.issn.1004-1524.2018.06.12

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In the present study, a pot experiment with Photinia frasery was conducted to examine NaCl addition (0, 0.1%, 0.2% 0.3%, 0.4%) on fresh weight of aboveground parts, plant height increase, plant growth, Na and K concentration, electrolyte leakage, relative water content, contents of proline, H₂O₂, malondialdehyde (MDA), anthocyanin and activities of the enzymes related to antioxidant system of plant leaves. It was shown that the growth of Photinia frasery was inhibited when NaCl addition amount was 0.2% or higher. Na and proline content, electrolyte leakage were significantly (P<0.05) increased, while K⁺/Na⁺ and relative water content were significantly (P<0.05) decreased. Salt stress induced oxidative stress of leaves. The H₂O₂ and MDA contents were significantly (P<0.05) increased by 162% and 128% in new leaves, and by 114% and 33% in old leaves, respectively. For new leaves, activities of SOD and CAT were significantly (P<0.05) enhanced, while POD activity and anthocyanin contents were significantly (P<0.05) decreased as the salt stress increased. However, activities of SOD and CAT were significantly (P<0.05) decreased, while POD activity and anthocyanin contents were significantly (P<0.05) increased as the salt stress increased in old leaves. Thus, it could be concluded that anthocyanin was an important antioxidant molecular of Photinia frasery, which partially replaced the antioxidant roles of CAT under salt stress.

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Effect of gibberellin on fruit quality and anatomical structure of Hongdeng sweet cherry

XUE Wanwan, GONG Ronggao, DING Jianlin, LI Keqiang, ZOU Jin, LI Rulong, PENG Honggui

2018, 30(6): 978-984. DOI: 10.3969/j.issn.1004-1524.2018.06.13

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In the present study, the effect of gibberellin (GA₃) on fruit quality and anatomical structure of Hongdeng sweet cherry was explored. It was shown that application of 60 mg·L^-1 GA₃ increased single fruit weight and longitudinal diameter. Application of GA₃ had no significant effect on fruit diameter, decreased the firmness of fruit, contents of soluble sugar, vitamin c and soluble solid, yet increased titratable acid content, which led to the overall decline in fruit quality. Anatomical structure analysis showed that with the increased concentration of GA₃, the thickness of cuticle and epidermis and firmness of fruit decreased, yet the size of flesh cells and epidermal cells become larger. It was suggested that the application of GA₃ should be reduced in production, especially for the application with high concentration (100 mg·L^-1).

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Effect of different concentrations of methyl-jasmonate on fruit quality of Chunmi peach

LI Qiuli, GAO Dengtao, WEI Zhifeng, WANG Zhiqiang, LIU Junwei, YANG Wenjia

2018, 30(6): 985-991. DOI: 10.3969/j.issn.1004-1524.2018.06.14

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Early maturing peach Chunmi was treated with different concentrations of methyl-jasmonate (MJ) to discuss its effect on fruit quality. Chunmi peach treated with of 125, 250, 500, 1 000 and 2 000 mg·L^-1 MJ at S3 stage were taken as the test materials to study the changes of single fruit weight, longitudinal and transverse diameter, soluble solid content, titratable acid content, firmness and color parameters of fruit during fruit development and ripening stage. The results showed that the single fruit weight, longitudinal and transverse diameter of Chunmi peach were significantly increased in 250 mg·L^-1 MJ treatment, were decreased significantly in 1 000 and 2 000 mg·L^-1 MJ treatment, while there was no significant difference in 125 and 500 mg·L^-1 MJ treatment compared with the control group. Different concentrations of MJ treatment increased the soluble solid content, reduced titratable acid content, but showed no significant difference among treatments. The firmness of fruit was increased in 250 and 2 000 mg·L^-1 MJ treatment, while there was no significant difference by the other MJ treatments compared with the control group. The color parameters showed no significant difference in all MJ treatments compared with the control group. The results of single fruit weight, longitudinal and transverse diameter, firmness and color parameters of fruit during fruit development period were consistent with that during fruit mature period. Different concentrations of MJ treatment increased the ratio of soluble solids to acidity, but there was no significant difference among treatments during fruit mature period. The fruit shape index, soluble solid content and titratable acid content showed no significant difference in all MJ treatments compared with the control group during fruit mature period. In conclusion, the 250 mg·L^-1 concentration of MJ was shown better application effect on the early maturing peach Chunmi during fruit development and ripening stage.

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Preliminary research on wound signal in fresh-cut bamboo shoots

HE Xiong, XIAO Shangyue, JIN Tunan, LUO Haibo, SUN Jincai

2018, 30(6): 992-998. DOI: 10.3969/j.issn.1004-1524.2018.06.15

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In order to explore the wound signal induced by mechanical damage in fresh-cut bamboo shoots, the effect of fresh-cut treatment on the damage signal transmission of bamboo shoots at different distances from the cutting site and the influence of methyl jasmonate (MeJA) on the synthesis MeJA and transmission of signal molecule in fresh-cut bamboo shoots were investigated with the exogenous signal molecule and its synthetic inhibitor salicylhydroxamic acid (SHAM). The results showed that the wound signal substance was generated at the cleavage site, and it could be transmitted to the adjacent uninjured tissue to increase the activities of phenylalanine ammonia lyase (PAL) and peroxidase (POD). MeJA treatment could increase the activities of PAL, POD, superoxide dismutase (T-SOD) and ascorbate peroxidase (APX) in fresh-cut bamboo shoots, while SHAM treatment had a certain inhibitory effect. Thus, it was hypothesized that MeJA could induce and participate in the synthesis and transmission of some damage signal after mechanical damage in fresh-cut bamboo shoots.

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MoSOK1, a putative germinal center kinase encoding gene, is required for fungal growth, conidiation and pathogenicity in Magnaporthe oryzae

FENG Xiaoxiao, LI Haijiao, LI Ling, WANG Jiaoyu, LIN Fucheng, LU Jianping

2018, 30(6): 999-1007. DOI: 10.3969/j.issn.1004-1524.2018.06.16

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SOK1 is a Ste20 protein kinase of the germinal center kinase (GCK) that is activated by oxidant stress and chemical anoxia. It is unknown so far whether SOK1 play roles in regulation of the fungal pathogenicity. Herein, we functionally characterized the SOK1 homologue in the rice blast fungus Magnaporthe oryzae(MoSOK1). MoSOK1 was up-regulated during conidial germination and appressorial formation. Targeted gene replacement showed that MoSOK1 acts as a regulator to fungal development and virulence in M. oryzae. Δmosok1 mutants showed decreased vegetative growth and conidiation, delayed conidial germination and a significant reduction in virulence. MoSOK1 was also found related to the sexual reproduction of the fungus. These data implicate that SOK1 signaling is required for fungal development and pathogenicity in phytopathogenic fungi.

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Synergetic inhibition against cotton Verticillium wilt disease of arbuscular mycorrhizal fungi cooperated with Bacillus vallismortis

ZHANG Guoyi, CHENG Lin, HUANG Liying, QIAN Qihui, GUAN Linlin, YE Wenling, LU Hongjuan, RAN Wei, ZHANG Ligan

2018, 30(6): 1008-1015. DOI: 10.3969/j.issn.1004-1524.2018.06.17

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In the present study, the cooperative effect of arbuscular mycorrhiza fungi (AMF) Glomus versiforme and Bacillus vallismortis HJ-5 against cotton Verticillium wilt disease was explored by greenhouse pot experiment, and the possible underlying mechanisms was revealed by high performance liquid chromatography (HPLC) and tablet test. As compared with control, AMF and HJ-5 showed synergistic inhibition against Verticillium wilt disease, as the disease index was reduced by 50.82%, root weight was increased by 125.00%, and the colonization of HJ-5 in rhizosphere was enhanced by 23.80%. HPLC analysis showed that HJ-5 could produce antifungal substances, i.e. iturin A and surfactin. Dual inoculation significantly increased the concentration of iturin A and surfactin in rhizosphere than single inoculation of HJ-5 by 13.38% and 11.27%, respectively. It was hypothesized that the main mechanism of synergetic inhibition was that AMF promoted cotton root growth, helped HJ-5 colonized in rhizosphere, and produce more antifungal lipopeptides.

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Controlling effects and mechanisms on rice diseases, pests and weeds in rice-based coculture system

WANG Yuejun, HUAI Yan, XU Jianfeng

2018, 30(6): 1016-1021. DOI: 10.3969/j.issn.1004-1524.2018.06.18

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Although the application of chemical pesticides has greatly increased agricultural productivity in modern agriculture, it has also brought about environmental and other negative effects. The rice-based coculture system was demonstrated as a successful approach to rice pesticide reduction and green, efficient productive way. This paper discussed the controlling effects and mechanisms of representative rice-based coculture system on rice diseases, pests and weeds, and its effect on pesticide reduction. It is suggested that further studies should be strengthened on technology packages for culturing rice and fish to achieve sustainable development of agriculture in Zhejiang Province.

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Impact of burning and water corruption of crop residues on environment

YUAN Weidong, LU Na, SONG Jiling, CHEN Qing, YAN Jing, WANG Weike, KANG Xueping, WANG Shiheng

2018, 30(6): 1022-1028. DOI: 10.3969/j.issn.1004-1524.2018.06.19

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In the present study, five materials, commonly used in mushroom substrates, i.e. Zizania latifolia leaves, rice straw, corn cob, mulberry branch and peels of Carya cathayensis Sarg, were selected as study objects to test the effects of two typical disposal methods (burning straightly and waste disposal in water) on environment. It was shown that burning (both open flame and smoulder) could produce a large amount of SO₂, fine particles (PM_2.5) and NO_x. In addition, smoulder produced greater amount of emission than open flame. The burning of Zizania latifolia leaves produced the largest amount of PM_2.5 and SO₂, which were 2 773.37, 3.96 mg·m^-3, respectively, while the burning of mulberry branches generated maximum amount of NOx, which was 61.5 mg·m^-3. Waste disposal (except peels of Carya cathayensis Sarg) decreased the pH of water body, yet increased the contents of suspended solids (SS), total nitrogen (TN), total phosphorus (TP), ammonia nitrogen, permanganate index and chromatic value in water. These indexes reached the maximum value in 30-50 d. Disposal of Zizania latifolia leaves in water led to 270.85 μg·L^-1 TN, which was 5 times greater than the others. The life cycle ‘multiplication-proliferation-death' of Escherichia coli was observed in contaminated water during 60 d and the amount of E. coli colonies reached to the highest level at 10 d, then died gradually. Both burning and disposal in water would lead to serious environment issues. Thus, it was suggested to improve the resources cyclic utilization of substrate residues.

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Determination of heavy metal contents and health risk evaluation of algae in coastal region of Zhejiang Province

CHEN Xingxing, WU Yue, ZHOU Chaosheng, LU Rongmao, ZENG Guoquan, HUANG Zhenghua

2018, 30(6): 1029-1034. DOI: 10.3969/j.issn.1004-1524.2018.06.20

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In order to investigate heavy metal pollution status in algae in Zhejiang Province and assess its risk to human health, inductively coupled plasma-mass spectrometry (ICP-MS) was used to detect the contents of Pb, Cd, As and Hg in three kinds of algae. The measured values were compared with current relevant standards in China. To anticipate the risks of heavy metal in algae, target hazard quotients (THQ and TTHQ) were calculated based on the health risk model proposed by the US Environmental Protection Agency (USEPA) in 2000. The results based on 34 samples showed that the standard-exceeding rates of Cd were 83.33% in Sargassum fusiform and 100% in porphyra. The standard-exceeding rates of Pb in Sargassum fusiform, Laminaria japonica, and porphyra were 16.67%, 44.44% and 100%, respectively, and the standard-exceeding rates of inorganic arsenic in Sargassum fusiform and Laminaria japonica were 100% and 66.7%, respectively. The three kinds of algae tended to be safe for adult, but posed a potential risk to children based on the health risks of single heavy metal pollution, especially the pollution of As in Sargassum fusiform and Laminaria japonica, and Cd in porphyra. Based on the assessment of health risks posed by multiple heavy metal pollution, these algae were found to be dangerous both for adult and children. The overall health risks of heavy metals were mainly due to As and Cd pollutions.

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Spatial characteristics and influencing factors of soil nutrients in hilly region of South China

ZHAO Yue, LUO Zhijun, ZHAO Jie, JIANG Chunyan, Cao Liping

2018, 30(6): 1035-1043. DOI: 10.3969/j.issn.1004-1524.2018.06.21

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The spatial characteristics of soil nutrients and their correlation with influencing factors were studied in hilly areas of southern China, to provide references for the efficient utilization of land resources and soil nutrient conservation. In the present study, Xiajiang County, Jiangxi Province was selected as the research object, soil samples were collected, and soil organic matter, available phosphorus, available potassium and alkali hydrolysable nitrogen contents were measured, and factors on soil nutrients were examined via spatial variability analysis and spatial autocorrelation analysis to explore the main influencing factors. It was shown that the distribution of soil nutrients in Xiajiang County was in accordance with the normal distribution, and the content of alkali hydrolyzable nitrogen was abundant. The contents of organic matter and available potassium were in the middle level, yet the available phosphorus content was low. The organic matter, available phosphorus showed moderate variation, yet available potassium showed strong spatial variability, alkaline hydrolyzable nitrogen showed weak spatial variability. Generally speaking, soil nutrients in the cultivated land in the west of the study area were generally lower than that in the east, and were higher in the area with higher elevation, or lower gradient, slope variability and topographic relief. Besides, soil nutrients were higher in the area with sandy rock weathering materials soil parent material, and were lower in the area with acid crystalline rock weathering substance as soil parent material.

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In situ simulated remediation of polycyclic aromatic hydrocarbons by activated sodium persulfate system

ZHANG Hongling, LI Sen, ZHANG Yang, ZHAN Sheng, SHANG Zhaocong

2018, 30(6): 1044-1049. DOI: 10.3969/j.issn.1004-1524.2018.06.22

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A system of activated sodium persulfate by sodium percarbonate was used to remove anthracene, pyrene, and benzoapyrene (PAHs) in soil,and the system complying with surfactants was applied in a simulation experiment of in situ remediation of PAHs. In the case that ratio of sodium percarbonate to sodium persulfate was 0.67,the maximum removal yield of PAHs reached 92.3%. On the basis, as the ratio of surfactants to system was 2%, the obtained highest removal rate of PAHs was 97.4%. Meanwhile, the effect of sodium percarbonate activated sodium persulfate combined with surfactants system to repair PAHs in soil in situ was significant, and the removal efficiencies of PAHs in upper and lower soils were both above 97.0% after 168 h.

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Effects of drying methods on quality of Sedum aizoon

YANG Ying, HAN Yanchao, GAO Haiyan, CHEN Hangjun, WU Weijie, DENG Shanggui

2018, 30(6): 1050-1055. DOI: 10.3969/j.issn.1004-1524.2018.06.23

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In the present study, three drying methods, i.e. hot air, vacuum, vacuum freezing, were applied to deal with Sedum aizoon. The color, and contents of vitamin C (V_C), total phenols, chlorophyll, protein, total flavonoids and oleanolic acid of the processed Sedum aizoon powder were determined to explore the effects of different drying methods on nutrition and functional components. It was shown that color, and the contents of Vc, total phenols, chlorophyll, protein, total flavonoids and oleanolic acid in the produced powder by vacuum freeze were significantly (P<0.05) superior than those in other products. Contents of protein and oleanolic acid in the powder produced by hot air were significantly (P<0.05) higher than those produced by vacuum. Based on the nutritional quality and functional value of Sedum aizoon powder, it was suggested that vacuum freeze was the optimal drying method.

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Dynamics of main nutritional components and aroma components of three sweet potato varieties during storage

LI Chen, XUE Guanwei, HUANG Jingyan, CUI Peng, LU Guoquan

2018, 30(6): 1056-1062. DOI: 10.3969/j.issn.1004-1524.2018.06.24

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In this paper, sweet potato cultivars Xinxiang, Zheshu75 and Nonglin54 were used as experimental materials to study the dynamics of main nutrients and aroma components of sweet potato during storage, to provide theoretical basis for the storage and preservation of sweet potato. The storage condition was set as temperature (14.95±1.27) ℃, and relative humidity (76.68±4.98)%. Samples were collected on 0, 20, 40, 60, 90, 110 d, respectively. It was shown that the dry matter content of three sweet potato cultivars increased, whereas the starch content and carotenoids showed a decreasing trend during storage. The number of sweet potato aroma components also decreased. The maximum content of aroma components in Xinxiang, Zhejiang75and Nonglin54 appeared on 40, 110, 20 d, respectively. Benzaldehyde and phenylacetaldehyde were not only the shared aroma components of three cultivars, but also the important components of sweet potato aroma.

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Preparation of magnetic molecularly imprinted polymer and detection of chloramphenicol in foods

LI Zengwei, WANG Na, LEI Chan, ZENG Yue, FU Zhenzhen, HE Li, LIU Shuliang, ZHOU Kang, AO Xiaolin, CHEN Shujuan

2018, 30(6): 1063-1072. DOI: 10.3969/j.issn.1004-1524.2018.06.25

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In the present study, magnetic separation method was combined with surface molecular imprinting. The magnetic molecularly imprinted polymer (MMIP) for chloramphenicol (CAP) was prepared by suspension polymerization method based on modified Fe₃O₄ nanoparticles as support material, CAP as template molecule, methacrylic acid (MAA) as functional monomer, ethylene glycol dimethacrylate (EGDMA) as corss-linker and azobisisobutyronitrile (AIBN) as initiator. Meanwhile, the magnetic molecularly non-imprinted polymer (MNIP) was prepared accordingly with the absent of CAP. Each step of the products were characterized by Fourier transform infrared spectrometer, microscopic morphology of MMIP and MNIP were observed by scanning electron microscopy, and the saturation magnetic strength of magnetic nanoparticles and MMIP were detected by vibrating sample magnetometer. The adsorption capacity of MMIP and MNIP were studied by binding experiments. Methodology evaluation was performed for detecting chloramphenicol in foods by using MMIP as solid phase extractant. The obtained MMIPs were 400-700 nm in diameter and had good monodispersity. It could be rapidly separated under an applied magnetic field. The MMIP had high maximum apparent adsorption capacity which was up to 29.18 mg·g^-1 and good selectivity. It had high recovery (86.30%-94.21%), high precision (RSD≤1.53%), good stability (RSD≤1.87%) and low detection limit (3.0 μg·kg^-1), when MMIP was used as solid phase extractant. Thus,the MMIP could be used to detect the chloramphenicol residue in food.

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Identification of navel orange lesions leaves based on parametric exponential non-linear residual neural network

YANG Guoliang, XU Nan, KANG Lele, GONG Man, HONG Zhiyang

2018, 30(6): 1073-1081. DOI: 10.3969/j.issn.1004-1524.2018.06.26

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In order to better identify navel orange leaf lesions. The paper proposed a parametric exponential nonlinear function asactivation function to improve the residual network and provided a new method of deep learning to identify lesions of navel orange leaves, which reduced the linear loss of rectification and improves the training effect. In this paper, CNN was used as a training tool, and navel orange leaf images were selected as training samples to discriminate the four types of diseased, deficient, normal and non-species. This new identification method had the great advantage of being convenient to use and more accurate for the identification of traditional plant diseases, and finally reached the accuracy rate of 97.18%-98.86%.

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Study on technical efficiency of large-scale pig breeding in China

LENG Bibin, JI Xueqiang, ZENG Hao, TU Guoping

2018, 30(6): 1082-1088. DOI: 10.3969/j.issn.1004-1524.2018.06.27

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In the present study, the panel data and data development analysis (DEA) and non-parametric frontier model were applied to measure large-scale pig farming comprehensive technical efficiency in all regions of China within 2008-2015. The comprehensive technical efficiency was decomposed and decomposition efficiency was analyzed. The results showed that comprehensive technology efficiency in western China was the highest, and comprehensive technology efficiency in eastern and central China should be improved. The influence of pure technical efficiency was higher than that of scale efficiency. Comprehensive technical efficiency of large-scale pig farming revealed by using stochastic frontier approach (SFA) model was compared with technical efficiency revealed by DEA, and the common points of the two methods were obtained. Based on the study conclusions, according suggestions for improvement of large-scale pig breeding in China were put forward. The advantages of large-scale breeding should be full played and clean breeding technology should be introduced in western China, the development of pig breeding should be promoted in the direction of automation and information in eastern China, the position advantage should be played in the central China, and technology exchange platform should be established in the central and eastern regions, and the fast logistics channel should be built in the central and western regions.

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Beekeepers' perception of climate change and its influencing factors: a case study of Sichuan Province

XU Liling, YOU Liangzhi, SUN Zhanli, ZHANG Shemei

2018, 30(6): 1089-1098. DOI: 10.3969/j.issn.1004-1524.2018.06.28

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Based on questionnaire data on beekeepers in five counties in Sichuan Province, we use model of proactive adaptation to climate change (MPPACC) framework and ordered Probit model to analyze the beekeepers' perception of climate change and its influencing factors. The results showed that the sensitivity perception and the severity perception of climate change of the beekeepers were stronger, while the efficacy perception and self-efficacy perception were weaker and the perceived adaptive cost was higher. Factors such as acquisition and accesses to free weather information, communication frequency to other beekeepers, the number of beekeepers in the family etc. have significant influence on beekeepers' perception of climate change. Especially, acquisition to free climate change information and the sensitivity perception, adaptive efficiency perception, self-efficacy perception and adaptive cost perception were the most significant factors. Finally, considering the needs to broaden beekeepers' accesses to climate information, we put forward suggestions to improve beekeepers' perception and response capacity towards climate change, such as encouraging participation of beekeepers' cooperatives and promoting communications among them.

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