Acta Agriculturae Zhejiangensis

Cloning and analysis of OsCSP2 gene promoter in rice (Oryza sativa L.)

XIAO Wenfei, NI Shen, QIU Jieren, WANG Shuzhen, XIN Ya, RUAN Songlin

2017, 29(6): 857-863. DOI: 10.3969/j.issn.1004-1524.2017.06.01

Abstract ( 862 )

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To understand functional expression profiling of cold shock protein gene OsCSP2, the promoter sequence and expression of OsCSP2 in rice were investigated. The promoter of OsCSP2, named Poscsp2, was cloned from the salt tolerant rice cultivar Shanyou 10 using PCR. An expression vector containing the Poscsp2::GUS fusion gene was constructed and introduced into rice. The expression patterns of Poscsp2 under low temperature and high salt stress were analyzed by GUS histochemical staining and qRT-PCR. The results showed that Poscsp2 was predicted to contain a large number of cis elements associated with low temperature, high salinity and other stresses. Expression levels of GUS reporter gene and OsCSP2 were significantly increased after low temperature and high salt stress treatment, indicating that Poscsp2 can improve expression levels of downstream genes under abiotic stresses.

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Effects of water stress on yield and water use efficiency of near isogenic introgression lines of high-quality indica rice Chuanxiang 29B

ZHANG Hong, ZHU Conghua, LI Xingyue, LI Qiyong, YANG Wenyu, REN Guangjun

2017, 29(6): 864-873. DOI: 10.3969/j.issn.1004-1524.2017.06.02

Abstract ( 898 )

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By pot experiments in greenhouse, the change characteristics in photosynthetic physiology, spike traits, grain traits, yield and yield components, and water use efficiency of near isogenic introgression lines were investigated, the relationships among the decreasing range of these indexes and drought tolerance of rice were analyzed under different water stress conditions. It was benefit to find the drought resistant material and the drought resistance identification index. The results showed that leaf net photosynthetic rate, spike length, neck-spike node, grain length, effective spike per plant, grain number per spike, seed setting rate, 1 000-garin weight, water use efficiency and drought resistance index of all the materials were significantly decreased with the increasing of the degree of water stress from seedling to maturity. In treatments T1 and T5, there were no significantly differences in grain yield of these tested materials. In treatments T2, T3 and T4, comparing with other materials, net photosynthesis rate, number of primary branches per spike, effective spike per plant, grain number per spike, seed setting rate, 1 000-grain weight, grain yield, harvest index and water use efficiency of C2 and C3 were much higher. In treatments T3 and T4, the drought resistance index of near isogenic introgression lines was significantly positively correlated to decreasing range of spike length and neck-spike node, and negative correlated to decreasing range of effective spike per plant, seed setting rate and water use efficiency. In summary, the drought resistance ability of C2 and C3 were significantly higher than any of other materials, and the higher yield could also be maintained under water stress. In the treatment T3 (0~30 cm soil water potential was -0.040~-0.045 Mpa) or T4 (0~30 cm soil water potential was-0.060~-0.065 Mpa), it was more accurate that the decreasing range of effective spike per plant, seed setting rate and water use efficiency were used as indicators for screening new material or combination with much stronger drought tolerance from seedling reviving to maturity.

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Cloning and functional characterization of a Na⁺/H⁺ antiporter gene from mulberry

LIU Yan, JI Dongfeng, SHEN Guoxin, WEI Jia, LIN Tianbao, ZHU Yan, LYU Zhiqiang

2017, 29(6): 874-881. DOI: 10.3969/j.issn.1004-1524.2017.06.03

Abstract ( 596 )

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Salinity is a major factor that adversely affects the growth and yield of plants worldwide. Plants Na⁺/H⁺ antiporters (NHX) play significant role in adaption to salt stress by compartmentalizing excess cytosolic Na⁺ into vacuoles and maintaining ionic equilibrium. Here we cloned an orthologous of NHX1 gene from You1#, a popular mulberry variety used especially in Southwest China. The length of MaNHX cDNA was 2 016 bp with a deduced molecular mass of 59.28 ku and pI 6.67. The consensus amiloride binding motif (⁸²LFFIYLLPPI⁹¹) was observed in the third putative transmembrane domain. Heterologous expression of MaNHX in Arabidopsis enhanced plants salt tolerance, which was verified by retaining higher chlorophyll content, higher antioxidase activity, slower reudction of malondialdehyde (MDA) content in transgenic lines than those in WT plants when confronted with the NaCl stress. Consequently, our work suggested that MaNHX was a candidate salt tolerance determinant gene in future mulberry breeding.

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Polymorphism of Nramp1 intron 6 and its correlation with piglet diarrhea

GU Shaobo, DU Qian, SHU Dan, LI Zhixia, TIAN Xiaojing, LIU Lixia

2017, 29(6): 882-887. DOI: 10.3969/j.issn.1004-1524.2017.06.04

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In order to research the polymorphism of Nramp1 intron 6 and its correlation with piglet diarrhea, this study analyzed the polymorphism of the Nramp1 intron 6 in Bamei pig and Yantai black pig by PCR-RFLP. The results showed that the polymorphisms of Nramp1 intron 6 were not rich in two pig breeds, two alleles (A, B) and two genotypes (AB, BB) were detected, the polymorphic information content was 0.33 in Bamei pig and 0.36 in Yantai black pig. The Chi square fit test showed that the Nramp1 intron 6 deviated from Hardy-Weinberg balance extremely in two pig breeds (P<0.01). The results of the least square showed that, the diarrhea resistance ability of Bamei pig was stronger than that of Yantai black pig (P<0.05). The gender and genotype of the Nramp1 intron 6 did not significantly influence piglet diarrhea alone (P>0.05), but the interaction of the gender and genotype influenced piglet diarrhea significantly (P<0.05) in female piglets, and the diarrhea score of genotype AB was greater than that of genotype BB (P<0.01), while in male piglets, these two genotypes had not significant difference in diarrhea score (P>0.05).

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Drug resistance spectrum analysis and MLST genotyping of Escherichia coli from refrigerated chicken in three regions of Zhejiang Province

YANG Hua, HE Xiangxiang, XIAO Yingping, QIAN Mingrong, ZHANG Qiaoyan, TANG Biao

2017, 29(6): 888-893. DOI: 10.3969/j.issn.1004-1524.2017.06.05

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Escherichia coli is a common opportunistic pathogen in food. Its drug resistance is severe in which the related resistance genes can easily spread. In this study, 59 strains of E. coli from refrigerated chicken in three regions of Zhejiang Province were isolated. Using drug resistant spectrum determination, 24 types of drug resistant spectrum were found, which showed that the drug resistant types of E. coli isolates were multifarious. Meanwhile, all the E. coli isolates were analyzed with MLST genotyping. The results showed that a total of 38 ST types were obtained, and two new ST types were found as well. On this basis, the phylogenetic analysis of 7 housekeeping genes showed a certain degree of the regional differences. This research results can provide scientific reference for the evaluation of drug resistance and traceability analysis of avian Escherichia coli in Zhejiang province, and a wider region in China.

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Isolation, identification of an ammonia-oxidizing bacteria and its application in treatment of wastewater from breeding of Macrobrachium rosenbergii

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

2017, 29(6): 894-902. DOI: 10.3969/j.issn.1004-1524.2017.06.06

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Ammonia-oxidizing bacteria play an important role in material cycles and water purifying processes in aquaculture ecosystems. In the present study, the screening of bacteria with capability to remove ammonia nitrogen from sludge on the bottom of an aquaculture pond was carried out, and the bacteria isolated were identified by biochemical tests and molecular methods, then, which were used for treatment of ammonia nitrogen in wastewater from breeding of Macrobrachium rosenbergii. The results showed that the isolated strain YX0703 with capability to remove ammonia nitrogen was Ochrobactrum anthropi, which was able to grow on both inorganic and organic nitrogen medium; Strain YX0703 not only had ammonia oxidation activity, but also had the ability to convert NO3- into NO2-, then convert NO2- into N₂. Moreover, strain YX0703 carried nitrite reductase K gene (nirK). The determination of minimal inhibitory concentration (MIC) indicated that strain YX0703 was resistant to cephalosporins, but sensitive to some antibiotics, such as quinolones, meropenem and tetracycline; the removal rate of ammonia nitrogen in wastewater from breeding of Macrobrachium rosenbergii were 96.98% and 97.44%, respectively, when shaker speed was 150 r·min^-1, at 25 and 30 ℃. These results suggested that the isolated strain YX0703 was capable of using inorganic or organic nitrogen, which also had activity of ammonia-oxidizing and denitrification, and was useful for removal of ammonia nitrogen in wastewater from breeding of Macrobrachium rosenbergii.

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Isolation, identification and antibiotic sensitivity of Streptococcus iniae from Pelteobagrus fulvidraco

YANG Yibin, XU Ning, YANG Qiuhong, LIU Yongtao, DONG Jing, AI Xiaohui

2017, 29(6): 903-909. DOI: 10.3969/j.issn.1004-1524.2017.06.07

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The study was aimed to investigate the cause about the outbreak of numerous deaths of Pelteobagrus fulvidraco at a fishery farm in Zhangzhou, Fujian Province, and to provide reference for the prevention and control of the disease for Pelteobagrus fulvidraco. The pathogenic bacteria were isolated and purified from liver, kidney and spleen of sick Pelteobagrus fulvidraco. And the identification of the strain si222 was conducted, using the biochemical identification and 16S rRNA gene sequence determination. The artificial infection test was performed, and the antimicrobial susceptibility test was conducted by disc diffusion method. The results showed that strain si222 was the pathogens of Pelteobagrus fulvidraco, the LD₅₀ of the isolate was counted to 2.63×10⁴ cfu·g^-1. According to morphological and biochemical characteristics as well as the result of 16S rRNA gene sequence analysis, the isolated strain si222 was Streptococcus iniae. Strain si222 was susceptible to oxacillin, cefadroxil, clindamycin and rifampicin and other 11 kinds of antibiotics. Meanwhile, it showed resistance to penicillin, amoxicillin, sulfisoxazole, florfenicol and other 6 kinds of antibiotics. The isolated strain si222 was the pathogenic bacteria to Pelteobagrus fulvidraco, and the disease can be prevented by administering drugs such as gentamicin and doxycycline in fisheries farming.

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Effects of glyphosate-resistant transgenic soybean on in vitro fertilization of male mice with reproductive damage

LU Chunbin, ZHANG Yan, CHEN Bohui, LIN Zebin, QIU Pingle, LIU Biao

2017, 29(6): 910-916. DOI: 10.3969/j.issn.1004-1524.2017.06.08

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To assess the effects of glyphosate-resistant transgenic soybean on reproductive function in mice with reproductive damage male mice were injected chemotherapeutical agent, cyclophosphamide (CP), to induced reproductive damage as model mice. The formation rate of zygotes, 2-cells, 4-cells, 8-cells as well as blastocysts and excellent embryo rate were evaluated in vitro fertilization after feeding with glyphosate-resistant soybean for 30 days. The results showed that no significant difference (P> 0.05) in the formation rate of zygotes, 2-cells, 4-cells, 8-cells, as well as blastocysts and excellent embryo rate were observed in cyclophosphamide (CP) treated reproductive-damaged mice fed with glyphosate-resistant soybean for 30 days, compared with the mice fed with traditional soybean. In conclusion, there was no adverse effect on the reproductive function in male mice after feeding with transgenic soybean for 30 days under CP induced pathologic conditions.

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Photosynthetic characteristics of 13 citrus cultivars

QIU Xia, YE Shuang, XIONG Bo, LIAO Ling, SUN Guochao, RONG Yi, LUO Jinyu, DAI Lin, WANG Zhihui

2017, 29(6): 917-925. DOI: 10.3969/j.issn.1004-1524.2017.06.09

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In order to study the photosynthetic characteristics of different citrus cultivars, 13 citrus varieties of 6 categories were used as test materials, and the instantaneous photosynthesis parameters, response characteristics to light, response characteristics to CO₂ and photosynthetic key enzyme activities were determined and analyzed. The results showed that there were significant differences in the photosynthetic parameters and photosynthetic enzyme activities among the 13 citrus cultivars. The light compensation point and the light saturation point were between 11.99-60.00 and 523.88-1 155.73μmol·m^-2·s^-1, respectively; the CO₂ compensation point and the CO₂ saturation point were between 20.01-72.02 and 1 364.44-1 811.29 μmol·mol^-1, respectively; the Rubisco, RCA and PEPC activity were respectively between 3.74-5.34, 10.97-23.83 and 5.06-9.39 μ IU, respectively. Comprehensive analysis of different citrus photosynthetic physiological parameters and photosynthetic enzyme activity showed that the photosynthetic capacity of Hejiang Bergamot, Huangguogan and Brazil Sour orange was strong; the photosynthetic capacity of Dayagan, Wanhong blood orange and Eureka lemon were the second; the photosynthetic capacity of Ehime Kashi.38 and Hongrou Guanxi pummelo were relatively worst. Higher activity of PEPC was detected in citrus, which is a key enzyme in the C₄ photosynthetic pathway, and the ratio of PEPC to Rubisco was between 0.95-2.26, indicating that there may be a weak C₄ photosynthetic pathway in citrus.

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Antioxidant capacity analysis in different sweet cherry varieties

GAO Fan, XIA Hui, WANG Xiu, WANG Jin, ZHOU Qiong, LYU Xiulan, LIANG Dong

2017, 29(6): 926-932. DOI: 10.3969/j.issn.1004-1524.2017.06.10

Abstract ( 615 )

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To investigate the International Commission on Illumination (CIE) values and antioxidant capacity of different varieties of sweet cherry fruit, Napoleon, Lapins, Sato Nishiki, Rainier, Hongdeng were used as materials to determine the content of the 3 antioxidants (total phenolics, total flavonoids and total flavanols).The antioxidant activities of 3 different mechanisms were evaluated in terms of the antioxidant activity of sweet cherry fruits, including ABTS^.+ radical scavenging capacity (ABTS), DPPH radical scavenging capacity (DPPH), and iron ion reducing ability (FRAP).The results showed that differences were found among different varieties. Red-fleshed sweet cherry varieties Hongdeng and Lapins owned higher a^* value and lower b^* value, while the yellow-fleshed varieties were on the contrary, and Napoleon had the highest L^* value. Hongdeng had the highest content of antioxidant compounds and also had the highest antioxidant capacity, while the yellow-fleshed varieties Sato Nishiki and Napoleon were the lowest. Moreover, significant or extremely significant positive correlations were found between total phenolics (TPC), total flavonoids (TFC) and each bioactive compound, and significant positive correlation was found between total flavanols (TFAC) and ABTS. All of these results suggested that red-fleshed sweet cherries contained high antioxidants, and Hongdeng is a promising hygienical function for human nutrition.

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Effects of embedding position of moisture sensors on growth characteristics of tomato in substrate culture in greenhouse

CAO Shaona, LI Jianshe, GAO Yanming, WU Suping, LIU Mengjin, LI Juan

2017, 29(6): 933-942. DOI: 10.3969/j.issn.1004-1524.2017.06.11

Abstract ( 718 )

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In order to study the effects of moisture sensor embedded position of real-time control irrigation system on growth characteristics of tomato in greenhouse, and to improve the precision and intelligence of water management in greenhouse substrate culture, the variety of Jingfan 301 was selected as the tested tomato, and single factor randomized block design of experiment was carried out with a total of 6 treatments and 3 replicates in each treatment. Using the same water upper and lower control irrigation, the effect of different treatments on growth indicators, photosynthetic indexes, quality, yield, dry matter, irrigation indicators and plant nutrients of tomato were analyzed. The results showed that T3 treatment (horizontal distance of 10 cm, vertical distance of 10 cm) had stronger root system, higher root to shoot ratio; and the accumulated values of net photosynthetic rate and transpiration rate were the highest, as well as the biggest plant fresh weight. T1 treatment (horizontal distance of 5 cm, vertical distance of 10 cm) had biggest plant nutrient accumulation. T4 treatment (horizontal distance of 10 cm, vertical distance of 15 cm) had highest light energy conversion. But, T3 treatment (horizontal distance of 10 cm, vertical distance of 10 cm) showed the highest ratio of sugar to acid and Vc content, and the production was up to 59 749.21 kg·hm^-2, significantly higher than other treatments. Water production efficiency in T3 treatment (horizontal distance of 10 cm, vertical distance of 10 cm) was the highest, which was up to 9.58 kg·m^-3 and better than that of T2 treatment (horizontal distance of 5 cm, vertical distance of 15 cm). According to the comprehensive analysis, for greenhouse tomatoes, the matrix moisture sensor at 10 cm from horizontal distance and 10 cm below the dripper was the suggested treatment.

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Effects on spectral indexes and fluorescence parameters of cowpea leaves after spraying cytokinin

HU Zhihui, ZHANG Liqin, WANG Yanjie, LAN Hong, GUO Rui, CHEN Gao, CHEN Chanyou

2017, 29(6): 943-950. DOI: 10.3969/j.issn.1004-1524.2017.06.12

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Ejiangdou 6, Ejiangdou 2, Ejiangdou 7 and Meiguo Didou were used as research materials and cytokinin (CTK) was sprayed in the budding period on cowpea leaves, with the same amount of water as a control (CK). Spectral indexes and fluorescence parameters of cowpea leaves were determined after spraying cytokininon at 7 days (early flowering stage), 14 days (flourishing flowering stage), 28 days (flourishing podding stage), 42 days (final stage). The results showed, with the increase of processing time, the change trends of CRI1 and ARI1 of all cowpea varieties leaves were down-up-down, while NDVI, WBI and PRI were up-down-up and there was no significant difference between spraying cytokininon and the control. On 28 days after spraying cytokininon, compared with the control, qP increased by 1.37%, Y(Ⅱ) and F_v/F_m reduced by 1.92%and 2.35%, respectively, which had no significant difference from CK.

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Comparison of chlorophyll fluorescence characteristics between two submerged macrophytes in Jiuzhaigou National Nature Reserve

GAO Li'nan

2017, 29(6): 951-958. DOI: 10.3969/j.issn.1004-1524.2017.06.13

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The Veronica undulata and Hippuris vulgaris in Jiuzhaigou were selected to examine diurnal variations of the maximum photochemical efficiency (F_v/F_m) and actual photochemical efficiency (Φ_PSⅡ) of PSⅡ, relevant parameters of rapid light curves (RLCs), photochemical quenching (qP) and non-photochemical quenching (NPQ) using pulse-amplitude modulated fluorometer (Junior-PAM) in Grass Lake. The purpose was to investigate the photosynthetic characteristics of these two submerged macrophytes and their adaptive strategies against environmental in alpine lakes. The results showed that the diurnal variations of Veronica undulata and Hippuris vulgaris of maximum relative electron transport rate (rETR_max) and half-saturation light intensity (E_k) presented typical unimodal curve. The F_v/F_m and Φ_PSⅡ of two submerged macrophytes showed V pattern diurnal cycle, F_v/F_m and Φ_PSⅡ were the highest at sunrise and sunset and the lowest at noon. The daily averages of rETR_max, E_k, qP and NPQ of Veronica undulata were significantly greater than those of Hippuris vulgaris. Therefore, Veronica undulata was macrophytes with strong capacity of light energy utilization and dissipation. A higher initial slope (α), chlorophyll b (Chlb) content and lower E_k in Hippuris vulgaris implied that it had higher capture light capacity and photosynthetic efficiency under weak light condition. In summary, significant differences in chlorophyll fluorescence parameters were found between Veronica undulata and Hippuris vulgaris, and Veronica undulata displayed more tolerance to strong light intensity than Hippuris vulgaris, it had a stronger ability for adaptation to local environment.

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The relationship between leaf surface anatomical structure and its function in four air plant species

FANG Minyan, ZHANG Ming, LI Jia, KONG Weiliang, DAI Dan, WANG Shanzhong

2017, 29(6): 959-965. DOI: 10.3969/j.issn.1004-1524.2017.06.14

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Tillandsia belongs to the air, rootless plants. The leaf surface and anatomical structure of 4 Tillandsia species (T. xerographica, T. streptophylla, T. velutina, T. tricolor) were studied. Results showed that: (1) Leaf surface of all the 4 varieties were covered with foliar trichomes, and shapes, cover forms and densities of foliar trichomes in 4 species were different. (2) The leaf epidermal cells were different in morphology. There were two cell shapes, near quadrangle and near quadrangle with peripheral wall. Cell sizes were not the same. There were no stomata on the upper epidermis, while had stomata on the lower epidermis. (3) The leaf anatomical structure included cuticular layer, palisade tissue, water storage tissue, vascular bundle and cavity, and no differentiation of palisade tissue and spongy tissue existed in mesophyll cells. A large amount of water storage tissue was near the upper surface or the lower surface, and there were big and obvious cavities between different vascular bundles. (4) The special leaf structure was closely related to the adaptability of the air grown and the different origins.

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Detoxification protocal optimization and virus detection in strawberry variety Hongyan

CHEN Ying, XIAO Chunlin, LUO Yanna, ZHANG Xiying, LIU Jiangna

2017, 29(6): 966-970. DOI: 10.3969/j.issn.1004-1524.2017.06.15

Abstract ( 634 )

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With strawberry variety Hongyan as the material, the sterile container seedlings were treated with high temperature treatment, low temperature freezing treatment combined with stem tip off detoxification. The double antibody step of serology and the clip art enzyme-linked immunoassay detection method were carried out to detect four kinds of virus. The survival rate and the detoxification efficiency of seedlings induced by meristem cells were compared. The results showed that the survival rate increased from 23.0% to 39.6%-60.0% under-1 ℃ cryogenic treatment with the stem tip stripping length increased from 0.3 mm to 0.5 mm, and all four virus were removed by the double antibody sandwich enzyme-linked immunoassay detection.

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Effect of Streptomyces JD211 fermentation products on the induced resistance to Magnaporthe grisea in rice

XU Zhirong, FU Yanhui, ZHAO Yingjie, WANG Ting, WEI Saijin

2017, 29(6): 971-976. DOI: 10.3969/j.issn.1004-1524.2017.06.16

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In order to study the effect of Streptomyces JD211 fermentation products on the induced resistance of rice, the leaves were induced by Streptomyces JD211 fermentation products and exhibited obvious resistance to Magnaporthe grisea. Rice seedlings were treated with different concentrations of Streptomyces JD211 fermentation products, and the dynamics of MDA contents and activities of defense enzymes in the rice leaves were measured. The results showed that the activities of CAT, POD and PAL in rice leaves could be improved by different concentrations of Streptomyces JD211 fermentation products, as well as the MDA content. The 20% Streptomyces JD211 fermentation products showed the most significant effect.

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Effects of biochar based fertilizer on seasonal variation of greenhouse gas emissions

CHEN Hongwei, HUANG Ling, FENG Lu, LI Xiaoqing, MENG Yutian, DAI Lin

2017, 29(6): 977-981. DOI: 10.3969/j.issn.1004-1524.2017.06.17

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A field experiment was performed to investigate the effect of application of biochar based fertilizer (0, 10, 20, 30 t·hm^-2) on soil N₂O,CO₂ emissions in 2015 and 2016. It was shown that compared with CK, CO₂-C emissions were increased in 2015 and 2016 by 0.81%-23.05% and 11.66%-30.94%, respectively, while, N₂O emissions were decreased by 13.32%-16.59% and 7.90%-19.57% in 2015 and 2016, respectively. Based on the results, although application of biochar based fertilizer increased soil CO₂ emission, it could effectively reduce soil N₂O emission, and decrease global warming potential (GWP) and greenhouse gas intensity (GHGI). Moreover, the effects of 30·t·hm^-2 application rate were sustainable between years.

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Improving quality of watermelon continuous cropping soil by reductive soil disinfestation

ZHOU Kaisheng

2017, 29(6): 982-987. DOI: 10.3969/j.issn.1004-1524.2017.06.18

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In the present study, the reductive soil disinfestation (RSD) was used to improve the quality of watermelon continuous cropping soil. The watermelon continuous cropping soil samples were incorporated with rice straw, ammonia, acetic acid, alcohol or different combinations, and then were used to test the effect of RSD treatment by planting watermelon seedling. It was shown that the pH values in soil samples added with acetic acid were significantly (P<0.05) lower than those in the samples of the original soil (OS) and controls (CK), but the pH values in other treated soil samples were all higher than those in OS and CK. The values of electrical conductivity (EC) increased in all treated soil samples. All treatments had no significant influence on soil bacteria. But, Fusarium oxysporum f. sp. niveum (FON), fungi and actinomyces were not detected in the soil samples added with ammonia, ammonia plus acetic acid, ammonia plus rice straw. The results of pot experiment showed that the number of death and mortality of watermelon in all treatment groups were lower than those in the control, as there were 3 groups with 0 mortality, and other 2 groups with mortality less than 7%, while the mortality rate in the control group was 43.3%. The watermelon growth momentum in all treatment groups was obviously better than that in the control group. The watermelon yields of all the treatment groups were also higher than that of control group, and there were 3 groups with significantly (P<0.05) higher watermelon yields than that of CK. It was concluded that RSD could effectively inhibit FON, alter soil microbial composition and structure, improve soil quality, and then control the obstacle of watermelon continuous cropping soil.

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Trends analysis of wastewater treatment used by constructed wetlands based on Web of Science statistics

CHENG Binbin, XU Linjun, MA Guilian, LIU Chang'e

2017, 29(6): 988-993. DOI: 10.3969/j.issn.1004-1524.2017.06.19

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Based on database provided by Web of Science, literature of the research results of wastewater treatment that had been used by constructed wetlands in recent 16 years were analyzed both in China and abroad in order to understand the development of wastewater treatment in constructed wetlands. The results showed that the publication quantity of international research findings was fluctuated with years from 2001 to 2016, and in general they were in an uptrend. The top five countries in terms of publication quantities were China, USA, Spain, Canada and Germany, among which China contributed the most, accounting for 21.1% and showing a great growth in publication quantities after 2007. Compared to western countries, China was low in citing rates of literature and in short of articles with high impact factors, which indicated the level and depth of China research remained to be further improved. This paper briefly analyzed the present research situation of wastewater treatment that had been used by constructed wetlands in China, and looked into the future development with positive anticipation.

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Rapid detection of chicken,duck and pork blending in beef products by multiple digital PCR

YANG Hua, WANG Xiaofu, XIAO Yingping, WEI Wei, XU Junfeng

2017, 29(6): 994-1000. DOI: 10.3969/j.issn.1004-1524.2017.06.20

Abstract ( 522 )

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In this study,a quickly detective method,using droplet digital PCR,for detection of beef,chicken,duck and pork was established. The results showed that the selected primers and probes for the four species had good specificity,and could quickly make out these four kinds of mean component in tested samples. In all of the digital PCR analysis, the fluorescence signals of FAM and HEX can be well distinguished. For detection of 9 samples,the numbers of droplets generated in the multiple digital PCR were all above 10 000, which could meet the requirements for the poisson formula analysis in digital PCR. meanwhile,the test results of the 9 samples using multiple digital were consistent with the known information of these samples.

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Synthesis technology and benzoylation cyanidin-3-glucoside catalyzed by lipase and structural characterization

YAN Zheng, LI Chunyang, HUANG Wuyang, ZHANG Lixia

2017, 29(6): 1001-1008. DOI: 10.3969/j.issn.1004-1524.2017.06.21

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The synthetic process of acylated cyanidin-3-glucoside catalyzed by enzyme was optimized through single factor experiment and response surface analysis according to the degree of conversion, and the product was identified by HPLC-MS and NMR. The optimum conditions were as follows: 41 ℃,-90 kPa, 43 mg·mL^-1 enzyme applied, concentration of cyanidin-3-glucoside 10 mg·mL^-1, concentration of methyl benzoate 0.4 g·mL^-1, reaction time 24 h. The structures of acylated cyanidin-3-glucoside products were identified as 3-(6″-benzoyl)-glucoside.

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Location of occluded apples based on gradient Hough transform

WU Qinggang, ZHANG Weiguo, CHANG Huawen, JIN Baohua, LIU Zhaoxia

2017, 29(6): 1009-1016. DOI: 10.3969/j.issn.1004-1524.2017.06.22

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In order to accurately locate the apples occluded by branches or other apples in natural environment, a method based on gradient Hough transform was proposed in the present study. Firstly, K-means clustering algorithm was used in Lab space to segment apples in natural environment. Secondly, morphological operations were made based on the segmented results to remove the influence of small area. Then, Sobel operator was adopted to extract the edge of apples. Finally, gradient Hough transform was applied to estimate the center and radius of target apples. Thus, the occluded apples could be automatically located. Experimental results showed that the method could effectively locate the occluded apples with the precision as high as 93.17%.

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Research on estimation approach of grassland vegetation coverage based on cellphone photo

DING Xiao, QIU Xinfa, GAO Jiaqi, GONG Jingyu

2017, 29(6): 1017-1025. DOI: 10.3969/j.issn.1004-1524.2017.06.23

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In the present study, 6 kinds of vegetation indexes (VEG, CIVE, EXG, EXGR, NGRDI, COM) were selected to estimate vegetation coverage of grassland based on RGB channel information, and their effects were compared with self-developed APP. Meanwhile, the influence on estimation error was discussed from 3 aspects including the green degree, weather conditions and vegetation coverage. Besides, the stability of each method was estimated from the perspective of threshold variation with illumination. It was shown that the estimation accuracy of 6 methods was higher than 90%. COM method reached the maximum accuracy of 95.41%, while, NGRDI method exhibited the lowest accuracy of 92.87%. The estimation error of each method for dark green grass was smaller than that of yellow green grass, and the estimation error in cloudy condition (cloudiness≥70%) was smaller than that in sunny condition (cloudiness≤10%). With the increase of vegetation coverage, the estimation error of VEG, CIVE, EXG, COM increased, while, no regular changes on estimation error of EXGR, NGRDI method were found. All 6 methods showed no obvious changes (threshold fluctuation≤0.02) with the changes of solar zenith angle and light intensity in a day. Under different weather conditions, the thresholds of VEG, CIVE methods had no obvious change (threshold fluctuation≤0.01), while, the thresholds of the remaining methods changed significantly (threshold fluctuation≥0.03). In short, all the 6 methods could satisfy the application in cellphone platform. As there was no need to consider the impact of illumination changes, VEG and CIVE possessed better versatility compared with the other methods.

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Research and design of intelligent control system for solar greenhouse based on ZigBee

YU Mingyue, WANG Lidi, LI Qingji, LI Yadi, LYU Tinghui

2017, 29(6): 1026-1036. DOI: 10.3969/j.issn.1004-1524.2017.06.24

Abstract ( 518 )

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In order to promote the modern management of greenhouse in North China, and to make the facility horticulture develop towards high yield and high efficiency, the intelligent control system was established based on ZigBee. To meet the needs of the control of agricultural greenhouse facilities, the system was produced by Jennic wireless microcontroller JN5139 as the control core, and the wireless sensor network was composed of sensor nodes and ZigBee wireless intelligent terminal. The system realized the real-time collection, monitoring, display, warning and control of the greenhouse environmental factors (including air temperature, humidity, light intensity, CO₂ concentration and soil pH value), and could provide the historical data of greenhouse environmental factors. In order to make the results more accurate, the sensor data sequence was smoothed three times on the nodes, then the smoothed data was sent to the gateway, and the linear regression analysis of data were carried out. The system basically met the needs of wireless, intelligent and precise modern facilities horticulture. The results showed that the greenhouse intelligent control system had the characteristics of stable and simple operation, and the results were accurate, which could effectively improve the management efficiency of the solar greenhouse, and had a good application prospect.

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Competitive situation analysis on patents regarding agricultural machinery engine fuel injection technology

WU Leiqi, SHEN Kai, ZHAO Zhijuan, YING Xiangwei

2017, 29(6): 1037-1042. DOI: 10.3969/j.issn.1004-1524.2017.06.25

Abstract ( 480 )

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Based on patent analysis into 3 kinds of diesel engine fuel injection system (electronically controlled unit pump system, electric control unit injector system, common rail fuel injection system), the competitive situation of these systems were investigated from 5 aspects, including patent application trend, technology development trend, regional distribution, rival agencies and applications in agricultural machinery engine. It was shown that the development of electronically controlled unit pump system and electric control unit injector system tended to be stable, with new technical inventions laying out in system components and new materials. Common rail fuel injection system related technologies were going to be mature. Germany and Japan hold absolutely leading positions in this field, while the research and development of China in this field was still in its infancy.

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Research on selection criteria of agri-products supply chain partner

ZHANG Cuiying, YOU Zhaotong, WANG Guoping

2017, 29(6): 1043-1049. DOI: 10.3969/j.issn.1004-1524.2017.06.26

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Establishing selection criteria of supply chain partner is the primary procedure of selecting copartner and the foundation of agri-products supply chain alliance. Furthermore, it contributes greatly to food quality safety control from its root and responds the changing market requirements. This paper analyzed features of agriculture market in China in order to seek the proper selection criterion system. The criterion system of copartner evaluation was constructed and index values were calculated by AHP. The results showed that the most important factors which affected agri-products supply chain partner selection were partner quality, performance capabilities and cooperative complementarity between two sides.

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Advance of research on microRNA function in cotton (Gossypium hirsutum)

LIU Yujiao, QIU Boyin, WANG Xiyan, XU Xiaojian, ZHU Shuijin, CHEN Jinhong

2017, 29(6): 1050-1056. DOI: 10.3969/j.issn.1004-1524.2017.06.27

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MicroRNAs (miRNAs) are important regulators of gene expression, and play essential roles in plant growth and development, response to adversity stress, etc. With the extensive application of deep sequencing technology, the function of miRNAs in cotton (Gossypium hirsutum) miRNAs has become a hot research topic. In this paper, the function of miRNAs in response to biotic and abiotic stress, fiber development and morphogenesis were reviewed. Eventually, the future research direction and emphasis were looked forward.

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