Acta Agriculturae Zhejiangensis

Genome-wide identification and expression analysis of soybean NIPs

WU Wei, FENG Zhijuan, XU Shengchun, LIU Na, ZHANG Guwen, HU Qizan, GONG Yaming

2018, 30(7): 1101-1109. DOI: 10.3969/j.issn.1004-1524.2018.07.01

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Nodulin 26-like intrinsic protein (NIPs) was one subclass of aquaporins which played vital roles in abiotic stress responses. Based on soybean genome database, a total of 13 GmNIP genes were identified on the genome level. Chromosome location analysis showed that these GmNIPs located on 11 chromosomes. Physical and chemical properties analyses revealed that the amino acid number of GmNIP proteins ranged from 261 to 296, with molecular masses of 27 to 32 ku and isoelectronic points of 6 to 10. Protein multiple alignments showed that all GmNIPs contained the typical regions of aquaporins: six putative trans-membrane domains (TM1 to TM6) and two NPA (Asp-Pro-Ala box) domains. Phylogenetic analysis showed that the classification of GmNIPs from soybean was consistent with Arabidopsis and rice NIPs. Gene structure analysis revealed that all GmNIPs were consisted of five exons and four introns. Promoter region analysis indicated that the upstream promoter region of most GmNIPs had stress and hormone responsive elements. Tissue expression analysis suggested that five GmNIPs had relatively high transcripts. Furthermore, the expression profile of GmNIP1;5 in response to drought stress was analyzed by the real-time PCR.

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Effects of DON and ZEA on IL-2, IL-4, IL-10 and IFN-γ of chicken spleen lymphocytes cultured in vitro

ZHANG Zhuo, GU Yanfei, YANG Shasha, WANG Yachao, DENG Junliang, REN Zhihua

2018, 30(7): 1110-1115. DOI: 10.3969/j.issn.1004-1524.2018.07.02

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The purpose of this study was to investigate the effects of DON and ZEA on the secretion of IL-2, IL-4, IL-10 and IFN-γ in cultured chicken spleen lymphocytes in vitro. The exposure doses of DON and ZEA were DON 0.012 5 μg·mL^-1 and ZEA 0.006 25 μg·mL^-1, DON 0.05 μg·mL^-1 and ZEA 0.025 μg·mL^-1, DON 0.2 μg·mL^-1 and ZEA 0.1 μg·mL^-1, DON 0.8 μg·mL^-1 and ZEA 0.4 μg·mL^-1, individually. Then, the effects of different doses of DON and ZEA on the concentrations and mRNA expressions of IL-2, IL-4, IL-10, and IFN-γ of chicken splenic lymphocytes in vitro were investigated. The results were as followed: The concentrations of IL-10 and IFN-γ, and the mRNA expressions of IL-2, IL-4, and IL-10 were increased with the DON and ZEA concentrations increasing (P<0.01). However, the concentrations of IL-2 and IL-4 were decreased with the DON and ZEA concentrations increasing (P<0.01). And the mRNA expressions of IFN-γ were increased at first and then decreased (P<0.01). DON combined with ZEA can affect the secretion of the IL-2, IL-4, IL-10 and IFN-γ with a dose-dependent. DON, ZEA combined with the spleen lymphocytes of the infected chicken to increase or decrease the level of inflammatory cytokines. The imbalance of Th1 and Th2 cytokines can result in cellular immune damage, which can be toxic to cells.

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Prevalence and genotyping of Giardia duodenalis in pre-weaned dairy calves in Sichuan Province

DAN Jiaming, OU Hongping, YAN Guangwen, TIAN Yinan, WEI Bin, TU Rui, XIAO Qicheng, TANG Li, YANG Tingyu, PENG Guangneng, WANG Yonggang, ZHONG Zhijun

2018, 30(7): 1116-1121. DOI: 10.3969/j.issn.1004-1524.2018.07.03

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To investigate the prevalence and molecular characteristics of Giardia duodenalis (G. duodenalis) in pre-weaned dairy calves in Sichuan Province, a total of 278 pre-weaned dairy calves' fecal samples were randomly collected in Sichuan. Samples were examined by the Sheather's sucrose flotation technique, and DNA was extracted. These samples were detected for Giardia by nest PCR amplification of β-giardin (bg), tpi and gdh genes. The results showed that 9.35% (26/278) was positive for G. duodenalis infection. One assemblage was detected, which was assemblage E. Multilocus genotyping showed that 7 subtypes of assemblage including 3, 2 and 2 new subtypes (bg: E13, E14, E16; tpi: E21, E24; gdh: E18, E19) were observed based on bg, tpi and gdh genes, respectively. The results indicated that assemblage E had the ascendant position in pre-weaned dairy calves in Sichuan Province and which showed its high genetic diversity.

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In vitro culture of CD103⁺ DCs from mouse bone marrow and effects of LPS on its morphology and functional characteristics

HOU Yanhua, ZHANG Kai, WANG Lei, SUN Jing, WANG Xurong, ZHANG Kang, WANG Xuezhi, LI Jianxi, ZHANG Jingyan

2018, 30(7): 1122-1131. DOI: 10.3969/j.issn.1004-1524.2018.07.04

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In order to establish a method of inducing and culturing CD103⁺ dendritic cells (CD103⁺ DCs) from C57BL/6 mice bone marrow in vitro,and investigate the effects of LPS on their morphology and functional feature. The bone marrow cells (BMCs) were isolated and cultured in the presence of recombinant granulocyte-macrophage colony-stimulating factor (GM-CSF) and fms-like tyrosine kinase 3 ligand (FLT3L), and then observed the effect of LPS on their morphology, phenotype and function. The results showed that the modality of BMCs diversified after being cultured in GM-CSF and FLT3L for 3 d, and grew clustered-liked.As time went on, the cell colonies became larger and larger. When culturing for 13 d, the BMC colonies started separating, typical morphology with dendritic processes could be observed. A number of morphologically typical dendritic cells were observed after culturing for 15 d. The dendritic structure became more obvious after BMCs were treated with LPS for 24 h under scan electron microscope. The induced BMCs were able to express CD103 on the cell surface, and the positive rate of CD103⁺DC was over 90%. The percentage of phagocytosis of CD103⁺ DCs was 25.70% in the group of RPMI-1640, but the percentage of phagocytosis in the group of LPS dropped to 10.33%. In the group of RPMI-1640, 41.31% CD103⁺DCs expressed MHC-II, 13.79% expressed CD83. The CD103⁺DCs treated by LPS showed a higher MHC-II expression rate of 68.10%, and increase expression rate in CD83 (24.71%). MTT assay showed that the ability of CD103⁺DC treated with LPS to stimulate the proliferation of the naive allergenic T cells was stronger than those CD103⁺DC untreated with LPS. In conclusion, a large number of dendritic cells can be generated by culturing BMCs from C57BL/6 mice in vitro,and LPS shows positive effect on CD103⁺ DC mature.

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Molecular characterization of Enterocytozoon bieneusi in captive giant panda, China

CAO Yuying, LI Wei, ZHONG Zhijun, XIAO Jie, GONG Chao, WANG Chengdong, LI Caiwu, YANG Haidi, HUANG Xiangming, WU Kongju, PENG Guangneng

2018, 30(7): 1132-1136. DOI: 10.3969/j.issn.1004-1524.2018.07.05

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In order to reveal the prevalence of Enterocytozoon bieneusi in captive giant panda and assess its zoonotic potential, we surveyed 31 fecal samples from captive giant panda based on nested PCR amplification of ITS gene of E. bieneusi. A total of 14 samples were positive for E. bieneusi, with an infection rate of 45.2%. Two genotypes were identified as SC02(13/14)and Peru6(1/14). Phylogenetic analysis revealed that these genotypes belonged to group 1b which had zoonotic potential. Giant panda could spread E. bieneusi to humans and lead to human microsporidiosis. The first report of SC02 in giant panda broadened the understanding of E. bieneusi of hosts.

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Ultramicroscopical histopathology and pathophysiology of “Shuibiezi” disease of Eriocheir sinensis

YANG Zongying, ZENG Liugen, WANG Yulan, YAO Yi, HOU Yujie, LEI Xiaoqing, XU Liangqing, XIONG Chunxian, YANG Xianle

2018, 30(7): 1137-1148. DOI: 10.3969/j.issn.1004-1524.2018.07.06

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Ultramicroscopical histopathological observations and pathophysiological analysis of Eriocheir sinensis with “Shuibiezi” disease were conducted by the transmission electron microscope technique and biochemical analysis. Ultramicroscopical histopathological results showed that pathological changes in different degrees were observed in gills, muscles and hepatopancreas of diseased crabs. The main pathological characteristics were degenerative and necrotic hepatic cells, decreased number of lipid droplets, and swelled or fractured microvillus. The swelled mitochondria showed abnormal appearance, such as swell, disorganization and reduction or vanish of the crista. The endoplasmic reticula were dilated or fragmented into lamellar structure. As for gills, the chitin layer became thinner, the cell nucleus of epithelial cells of gill was heterochromatinized, the finger shaped protuberance extended from the chitin layer was collapsed and disappeared, microvilli arranged disorderly and fractured or vanished, the sub-cuticular space was expended, the number of mitochondria was decreased and mitochondria was swelled or vacuolar with the decrease of crista, the number of lysosome was increased and lysosome formed autophagosomes with vacuoles and vacuolar mitochondria. Bacteria particles appeared in the severely changed gill epithelium, but no inclusions and other virions. The main pathological changes in muscle cells were relaxed and cracked muscular fibers, melted sarcoplasmic reticulum or formed many small bubbles, the number and volume of mitochondria decreased, and the nuclear was pyknosis and marginalized. Pathophysiological analysis results showed that glucose content and GOT activity in hemolymph of diseased crabs were higher than that of healthy crabs, ALP and GPT activities were significantly higher than those of healthy crabs, whereas triglyceride content was significantly lower than that of healthy crabs. As for hepatopancreas, the liver glycogen content of diseased crabs was significantly lower than that of healthy crabs, and ALP and ACP activities were lower than those of healthy crabs.

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De novo assembly and functional annotation of transcriptome data of Rhododendron pulchurum cv. Baifeng 4 leaf

WANG Hua, WANG Wangwei, WANG Dongliang, ZHANG Shihu, HU Xinfang, LU Shiyu, GONG Xuemei

2018, 30(7): 1149-1159. DOI: 10.3969/j.issn.1004-1524.2018.07.07

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In this study, the transcriptome analysis of two-years-old cuttage potted seedlings of Rhododendron pulchurum cv. Baifeng 4 were carried out by the second generation of Illumina sequencing technology. The results showed that 213 723 424 Clean reads and 53 568 Unigenes whose average length was 930 nt were obtained after filter, quality control and De novo assembly. Based on it, 28 877 Unigenes were successfully annotated to some databases, such as Nr, Swiss-Prot, KOG (clusters of orthologous group), KEGG (Kyoto encyclopedia of genes and genomes) (E-value≤10^-5). Comparing with the sequence homology of Nr database, it was found that Rhododendron pulchurum had higher homology with Vitis vinifera, but lower homology with other species. There were 30 508 Unigenes from KOG database and divided into 25 categories. GO (gene ontology) analysis showed that 11 928 Unigenes were identified as potentially involved in environmental stresses. From KEGG analysis, 6 478 Unigenes were assigned to 132 known pathways. Among these pathways, 384 Unigenes were related to biosynthesis and transduction of phytohormone. In addition, there were 1 062 Unigenes were annotated as transcription factors, a total of 8 738 SSR markers were excavated, which laid the foundation for further study of Rhododendron.

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Effects of various concentrations of EGCG on seed germination and resistance in cucumber under NaCl stress

LI Yang, LIU Kai, WEI Jipeng, ZHANG Lan, LI Xin, HAN Wenyan, LI Qingyun

2018, 30(7): 1160-1167. DOI: 10.3969/j.issn.1004-1524.2018.07.08

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The present study investigated the effects of exogenous application with various concentrations of EGCG (epigallocatechin gallate) on seed germination, malondialdehyde (MDA) content and activities of antioxidant enzymes in radicles of cucumber under NaCl stress. Results showed that, without salt stress, seed germination, radicle length and root branch number responded differently to various concentrations of EGCG. While those were significantly restrained under 200 mmol·L^-1 NaCl treatment, accompanying the great increases in content of MDA and activities of antioxidant enzymes. Furthermore, application with various concentrations of EGCG(10, 100, 1 000 μmol·L^-1)could significantly increase seed germination, radicle length and root branch number under NaCl stress. Moreover, EGCG application decreased MDA content by improving the activities of antioxidant enzymes (APX, CAT, SOD and POD) under NaCl stress. And the treatment with 100 μmol·L^-1 EGCG performed best. These results suggested that exogenous EGCG could enhance seed germination and mitigate NaCl-induced salt stress through promoting the activities of antioxidant enzymes in cucumber.

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Physiological response to heat stress and heat resistances evaluation of 14 Capsicum varieties

HU Nengbing, PANG Dandan, SUI Yihu, SHU Yingjie, HE Keqin, ZHU Xiaomei

2018, 30(7): 1168-1174. DOI: 10.3969/j.issn.1004-1524.2018.07.09

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Under the heat stress of 38 ℃, the heat damage index and chlorophyll fluorescence parameters of 14 Capsicum varieties at different processing time (24, 48, 72 and 96 h) were studied, and furthermore their abilities of heat resistance were evaluated and the best Capsicum varieties were selected by dealing with the data above. The results showed that, with the increase of high temperature treatment time, the thermal damage index(TDI) and water loss rate(WLR) of pepper increased and fluorescence parameter of F_v/F_m decreased. It was concluded by the membership analysis method and the parameters of chlorophyll fluorescence method by heat injury index that variety 2, 6, 1 can be used as the germplasm resisting high temperature stress and variety 11, 10 would be eliminated, with the coincidence rate reached 75% of the two methods, which could screen the optimum combination and verify the reliability of the two methods.

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Effects of substrates prepared from Dendrobium officinale cultivation waste and mushroom residue on growth, yield and quality of strawberry

MA Yisheng, HONG Chunlai, WANG Weiping, YAO Yanlai, ZHU Fengxiang, CHEN Xiaoyang, XUE Zhiyong

2018, 30(7): 1175-1181. DOI: 10.3969/j.issn.1004-1524.2018.07.10

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Strawberry pot experiments were performed on compound substrates, which contained different ratios of Dendrobium officinale cultivation waste (DOCW) or mushroom residue (MR), and the single component substrates, which contained either DOCW or MR, were designed as controls. It was shown that the compound substrates contained more abundant and balanced nutrient elements, and their physical and chemical properties, such as bulk density, electrical conductivity, porosity and pH value were more suitable for strawberry growth. There were no significant differences in nutritional growth of strawberry among different treatments. However, significant differences in reproductive growth such as fruit quality and strawberry yield were found among treatments. The yield of strawberry in treatment H5 (80% MR+20% soil) and H1 (80% DOCW+20% soil) were significantly (P<0.05) higher than the other treatments, and the contents of total soluble solid, vitamin C, soluble sugar, and the ratio of total soluble solid to titratable acidity in H5 were the highest among all the treatments. Therefore, the comprehensive cultivation effects of treatment H5 on strawberry were obviously superior, and it deserved promotion as compound substrate for strawberry cultivation.

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Selection of reference genes in wheat infected by Chinese wheat mosaic virus (CWMV)

WU Ne, CHEN Xuan, JIANG Yaoyao, ZHANG Tianye, YANG Jian, ZHU Tongquan, ZHANG Hengmu, CHEN Jianping

2018, 30(7): 1182-1187. DOI: 10.3969/j.issn.1004-1524.2018.07.11

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Quantitative PCR (qRT-PCR) has been widely applied to the expression analysis of functional genes. The selection of appropriate reference genes is required for the statistical analysis of qRT-PCR data. To select the suitable reference genes in wheat infected by Chinese wheat mosaic virus(CWMV), 10 housekeeping genes were screened by calculations of PCR efficiencies and melting curve analysis, suggesting that 8 out of 10 genes could be used as candidate reference genes. And then their temporal and spatial expression patterns were investigated by qRT-PCR in the different tissues and at the different stages of wheat inoculated with CWMV. These quantitative data were further evaluated by the computer programs geNorm and NormFinder in this study. The analysis results consistently revealed that two genes, 26S and CDC, were stably expressed in all tested tissues and CWMV-infected wheat samples, indicating that the combination of 26S and CDC could be selected as reference genes for expression analysis of functional gene during the interaction between CWMV and wheat.

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Pathogen identification and susceptibility to fungicides on tea anthracnose in Xinchang of Zhejiang Province

XIE Yunye, ZENG Sijin, YUAN Yue, WANG Lianping, FANG Li, WANG Hanrong

2018, 30(7): 1188-1193. DOI: 10.3969/j.issn.1004-1524.2018.07.12

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Tissue isolation method was used to isolate the pathogens from the symptomatic leaves of tea which were collected from Xinchang of Zhejiang Province. These isolates were identified into Colletotrichun gloeosporioides based on its morphology, pathogenicity and rDNA ITS sequence. Fungicide sensitivity for this pathogen was tested in this study. The results showed that C. gloeosporioides is more sensitive to 250 g·L^-1 pyraclostrobin, 65% zineb, 45% prochloraz, 250 g·L^-1 propiconazole and 70% thiophanate-methyl (EC₅₀<1 mg·L^-1), followed by 325 g·L^-1 difenoconazole·azoxystrobin, 10% difenoconazole, 430 g·L^-1 tebuconazole, 38% pyraclostrobin·boscalid and 10% hexaconazole (1 mg·L^-1<EC₅₀<10 mg·L^-1). The sensitivity of C. gloeosporioides to 12.5% diniconazole, 25 g·L^-1 fludioxonil and 42.8% fluopyram·trifloxystrobin were lower than the others (EC₅₀>10 mg·L^-1). Moreover, the mycelium of C. gloeosporioides can growth well with 50% carbendazim and 50% boscalid.

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Genetic characteristics and taxonomic classification of paddy field soils developed from two parent materials in East Hunan

MAN Haiyan, HUANG Yunxiang, SHENG Hao, YU Zhan, ZHOU Qing, YUAN Hong, ZHANG Yangzhu

2018, 30(7): 1194-1201. DOI: 10.3969/j.issn.1004-1524.2018.07.13

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Six typical paddy field soils, which were developed from either purple rock weathering or river and lake sediments in the East Hunan, were investigated to make its attribution in Chinese Soil Taxonomy clear. The diagnostic horizon and characteristics of 6 paddy field soils were retrieved out and the taxonomic classification were determined according to Chinese Soil Taxonomy (3rd edition) , based on soil forming environment, physical and chemical properties, and genesis characteristics of soil profiles. It was shown that the paddy soil developed from 2 tested parent materials exhibited similarity in soil pH, content of organic matter and total iron. With the deepening of soil profiles, soil pH increased, yet organic matter content decreased. The total iron content of topsoil was lower than that in plow pan and hydragric horizon. Soils developed from purple rock weathering were redder than that developed from river and lake sediments, and the soil particle composition of the former was similar between solum among soil profiles. Fe_o/Fe_d in anthrostagnic epipedon was higher than that in subsoils, while the (Fe_d-Fe_o)/Fe_o was the contrary. The determined diagnostic horizons of all test soils were anthrostagnic epipedon and hydragric horizon, and the diagnostic characteristics were mainly anthrostagnic soil moisture regime, gleyic features and redox features. The 6 paddy field soils belonged to stagnic anthrosol, and could be further divided into 3 groups and 3 subgroups, i.e. gleyi-stagnic anthrosols, Fe-accumuli-stagnic anthrosols, hapli-stagnic anthrosols, and typic gleyi-stagnic anthrosols, typic Fe-accumuli-stagnic anthrosols, typic hapli-stagnic anthrosols. Finally, 5 soil families and 6 soil series of test soils were established.

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Effects of straw-returning and tillage depth on soil properties in plough layer of paddy soil

HU Xinyi, FU Qinglin, LIU Chen, DING Nengfei, LIN Yicheng

2018, 30(7): 1202-1210. DOI: 10.3969/j.issn.1004-1524.2018.07.14

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To investigate the effects of different tillage and straw management on soil nutrients, enzyme activities and microbial community structure, a field experiment was conducted at two experimental stations in Lanxi and Jinhua in 2016.The experiment consisted of four treatments with three replicates arranged in a completely randomized design: shallow tillage (tillage depth 1012 cm) without straw, shallow tillage with straw incorporation,deep tillage (tillage depth 1820 cm) without straw and deep tillage with straw incorporation. Soil nutrients, enzyme activities, phospholipid fatty acid (PLFA) content and microbial community composition were determined. It was shown that addition of straw in topsoil increased the contents of soil alkali-nitrogen and Olsen phosphorus, but there were no significant differences in pH, available potassium and organic matter contents among treatments. Straw incorporation at the same tillage depth increased rice grain yield, but tillage depth with the same straw management had no significant effect on rice grain yield. Increasing tillage depth or addition of straw in topsoil increased the activities of soil acid phosphatase and β-glucosidase. There were no significant differences on Shannon index among treatments. Straw incorporation at the same tillage depth increased total PLFA, bacterial PLFA and fungal PLFA, but deep tillage with the same straw incorporation decreased fungal PLFA. Straw incorporation with proper tillage depth was an effective management measure to increase soil nutrients, soil enzyme activities, contents of soil microbes, and consequently grain yield.

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Prediction for spatial distribution of soil organic matter based on random forest model in cultivated area

YANG Yucen, YANG Lian'an, REN Li, LI Congli, ZHU Qun'e, WANG Tiantai, LI Xinyao

2018, 30(7): 1211-1217. DOI: 10.3969/j.issn.1004-1524.2018.07.15

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In the present study, the cultivated area of Zhouzhi County in Shaanxi Province was selected as study area with 192 soil samples collected, and the soil organic matter (SOM) content and distribution were predicted based on random forest (RF) model. The prediction accuracy was verified by 29 (15%) independent verification points, and the results were compared with ordinary kriging (OK) and cokriging (COK). It was shown that SOM contents in the training set and verification set were all moderately variable, and were classified into the medium low level. The SOM content was relatively high at east coast of Heihe River in middle and southern area, whereas was relatively low at Weihe River coast in the north-eastern area. By ranking the importance of variables, it was revealed that the main factors affecting the soil organic matter in the study area were elevation and rainfall. Compared with OK and COK, the correlation coefficient of prediction value and actual value of RF (0.782) was higher, yet the mean absolute error (0.618 g·kg^-1) and root mean squre error (2.062 g·kg^-1) were lower, which suggested that RF model yielded a more realistic spatial distribution of SOM.

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Preparation technology and properties analysis of biochar-based microbial fertilizers

SUO Guifang, LYU Haohao, WANG Yuying, LIU Yuxue, HE Lili, YANG Shengmao

2018, 30(7): 1218-1228. DOI: 10.3969/j.issn.1004-1524.2018.07.16

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In the present study, the rice husk biochar, NPK fertilizer, and Bacillus subtilis spores were prepared for the development of new biochar-based microbial fertilizers, by using three manufacture technologies including the mixed method, adsorption method and mixed granulation method. The content of rice husk biochar was set as 10%, 20%, 30%, respectively. The properties of these biochar-based microbial fertilizers, including the pH value, the total nutrients content, the elemental contents, the number of living bacteria, and nutrient release characteristics were determined. It was shown that biochar-based microbial fertilizer prepared with mixed granulation method had the best ability to absorb the NPK fertilizer and microbial spores, and was followed by those prepared by the adsorption method and the mixed method. The elements contents of 9 biochar-based microbial fertilizer samples were rich. The content of C and N increased while the content of H decreased with the increase of the addition amount of rice husk biochar under the same preparation condition. The pH value of the 9 biochar-based microbial fertilizer samples were acid or close to neutral, and the total nutrients contents met the requirement of the national standard for compound fertilizer. The number of living bacteria was 3.4×10⁵-7.0×10⁵ g^-1 after the spores of Bacillus subtilis were compounded with rice husk biochar and NPK fertilizer for 60 days. Under the same process conditions, the number of living bacteria of biochar-based microbial fertilizers increased with the increase of the addition amount of rice husk biochar. The seven days nitrogen accumulated water dissolution experiments showed that the 9 biochar-based microbial fertilizer samples had certain slow release function. The nitrogen release rate of biochar-based microbial fertilizers with the same preparation technology decreased with the increase of the addition amount of rice husk biochar. With the same content of rice husk biochar, mixed granulation method had the best slow release characteristics, and was followed by the adsorption method and the mixed method. Therefore, the biochar-based microbial fertilizer with 30% rice husk biochar made by mixed granulation method had optimal property, and the effect should be further verified by pot or field experiment and to promote its application in small scale.

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Screening, identification and characteristics of Lysinibacillus fusiformis 23-1 for petroleum degradation

LI Guoli, ZENG Xiaoying, ZHAI Lixiang, LENG Yan, LIU Mengyuan, LI Shiweng, CHEN Tuo

2018, 30(7): 1229-1236. DOI: 10.3969/j.issn.1004-1524.2018.07.17

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Petroleum spill has caused serious environmental problems and enormous economic losses. The microbial bioremediation have been shown to be the most effective way for the removal of petroleum pollution in environments. We isolated a biosurfactant producing and petroleum degrading bacteria from petroleum contaminated soil in the region of Qilian Mountain Baoping River in the Qinghai-Tibet Plateau. We further investigated the petroleum emulsifiability, degradation properties, and degradation conditions of the isolated strain, as well as the uses of hydrocarbons with different carbon chain by the isolated strain. A biosurfactant producing and petroleum degrading strain, assigned as 23-1, was successfully isolated on the medium with petroleum as sole carbon source using the blood plate screening method. Morphological and 16S rRNA gene sequence analysis identified this isolated strain as Lysinibacillus fusiformis strain 23-1. The colorimetric and ultrasonic analysis indicated that the emulsifiability and emulsion stability of strain 23-1 were 19.6% and 37.5%, respectively, suggesting that L. fusiformis strain 23-1 can reduce the oil-water interfacial tension and enhance the oil solubilization. The gravimetric method analysis showed that 57% petroleum added in the medium could be degraded by L. fusiformis 23-1after 8-d incubation, with pH 7.5 and 25 ℃. Further GC-MS measurement indicated that L. fusiformis 23-1 strongly degraded direct chain saturated alkanes, partly degraded the short chain alkanes and long chain alkanes, but weakly degraded the aromatic hydrocarbons. L. fusiformis 23-1 can produce certain biosurfactant and degrade petroleum during growth in the medium added with petroleum as sole carbon source. This strain could be used for bioremediation of petroleum contaminated environments.

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Patulin reduction by Lactococcus lactis strain

ZHU Ruiyu, HUANG Conghui, ZHU Yanjin, YOU Yuru, LI Ling, SUN Lü

2018, 30(7): 1237-1243. DOI: 10.3969/j.issn.1004-1524.2018.07.18

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In the present study, a strain of food-grade microorganism Lactococcus lactis MG1363 was screened for its ability of patulin degradation. The effects of environmental temperature and pH value on patulin degradation by L. lactis MG1363 were assessed, and the mechanisms of patulin degradation were also studied by bacterial cells inactivation and crude enzyme extraction. The results indicated that temperature and pH influenced patulin degradation significantly (P<0.05), and the optimal condition was 30 ℃, pH value 5.5-6.0. The mechanisms of patulin degradation by L. lactis MG1363 involved both physical adsorption and enzymatic degradation, and the degradation relevant enzymes were located inside the bacterial cells. The property of patulin-degrading by L. lactis MG1363 was constitutive, nevertheless, the enzymatic activity of patulin degradation was enhanced by low amount of patulin.

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Raman spectroscopic analysis and identification of Thunberg Fritillary Bulb

SUN Caixia, PAN Zhiyan, DAI Fen, ZHENG Weiran, YU Guoguang

2018, 30(7): 1244-1248. DOI: 10.3969/j.issn.1004-1524.2018.07.19

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Fresh samples of Thunberg Fritillary Bulb from 5 main planting areas in Zhejiang Province were collected and the laser Raman spectroscopy was used to analyze and identify the samples. The results were used for the characteristic peak and first-order derivative analysis. The results showed that several characteristic strong peaks could be found at 875,912,965,1 153,1 312,1 410,1 482,1 598,1 641 cm^-1 shift, and the affiliation of these peaks was identified. The results showed that Raman spectroscopic has the advantages of nondestructive, fast and simple, and the identified 9 peaks would be realized as the measures to distinguish the truth, origin area and traceability of Thunberg Fritillary Bulb.

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Microbial community structure and drug resistance gene on water, ground and surface of slaughtering equipment in poultry slaughterhouse

WANG Peipei, YANG Hua, DAI Xianjun, GUI Guohong, XIAO Yingping

2018, 30(7): 1249-1258. DOI: 10.3969/j.issn.1004-1524.2018.07.20

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The aim of this study is to investigate the microbial community structure and resistance genes on water, surface of floor and slaughtering equipment. The water and the bacteria collected on floor and surface of slaughtering equipment by sterile gauze swab were sampled in hanging, slaughtering, scalding, dehairing, eviscerating rooms of a large-scale poultry slaughterhouse in Zhejiang Province. The microbial community structure was analyzed based on 16S rRNA V3-V4 by Illumina Hiseq 2500, while the nine major categories of 24 resistance genes were tested by PCR method. The results showed that Proteobacteria, Firmicutes and Actinobacteria were the predominant phyla on water and the surface of floor and slaughtering equipment in the different areas of the slaughterhouse, their relative abundance were 27.55%-88.99%, 3.45%-59.78%, 4.35%-31.78%, respectively. The dominant genera included Acinetobacter, Psychrobacter, Thermus, Pseudomonas and Shewanella, accounting for 46% of the total taxa. The drug resistance gene detection rate of sulⅠ, sulⅡ and qnrS was 100%, while blaTEM, tetB and aadA1 was 94.4%, and floR and mecC was 88.9%. It was concluded that there were some conditional pathogenic bacteria and spoilage bacteria with many kinds of bacterial resistance genes on water and the surface of ground and slaughtering equipments in different areas of the slaughterhouse.

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Research progress of first-flower-node trait in pepper(Capsicum spp.)

YUAN Xinjie, FANG Rong, ZHOU Kunhua, ZHANG Hongyan, CHEN Xuejun

2018, 30(7): 1259-1266. DOI: 10.3969/j.issn.1004-1524.2018.07.21

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Pepper (Capsicum spp.) is an important economic crop in Solanaceae with sympodial shoot architecture. First-flower-node (FFN), also described as the number of leaves on the primary axis (Nle), number of leaves on primary stem until first flower or number of nodes, is a main maturity trait in pepper, which is closely related to precocity and early yield. Flowering time has an important role in crop domestication and adaptation to novel environments. The timing of the initial transition to flowering can be best measured by FFN. This paper reviewed the research of FFN in pepper, including genetic mapping, knowledge of associated genes identified by EMS induced mutants, functional conservation and diversity of these genes, as well as the interrelationship of them, and prospected the future directions and suggestions.

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Arbuscular mycorrhizal fungi as a potential indicator to assess effects of genetically modified crops on soil microorganisms

LIANG Jingang, JIAO Yue, LIU Pengcheng, ZHANG Xiujie

2018, 30(7): 1267-1272. DOI: 10.3969/j.issn.1004-1524.2018.07.22

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Genetically modified (GM) crops are closely related to the environmental elements of soil ecosystem, microorganisms are the important components of the soil ecosystem, thus, whether the large-scale plantation of GM crops would affect soil microorganisms has become an important aspect of ecological risk evaluation. There are many kinds of microorganisms in the soil, so it is necessary to select some representative microorganisms as indicators. As an important soil environment indicator, arbuscular mycorrhizal fungi (AMF) play an irreplaceable role in improving soil quality and healthy status, enhancing biological diversity and promoting plant growth. This paper reviewed the influence of main GM crops on AMF, and suggested that AMF could be used as a potential indicator to assess the effects of GM crops on soil microorganisms.

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