Acta Agriculturae Zhejiangensis

Cloning and expression analysis of a fibrillin gene in plant

JIANG Yaoyao, LI Jing, CAI Nianjun, CHEN Jianping, ZHANG Hengmu

2019, 31(9): 1399-1404. DOI: 10.3969/j.issn.1004-1524.2019.09.01

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Plant fibrillin (FBN), a large family of conserved proteins, is widely distributed in plant kingdom, but its biological properties and functions in Nicotiana benthamiana are still unclear. To analyze its expression and function, an FBN gene (NbFBN) was amplified and cloned from Nicotiana benthamiana by RT-PCR. Phylogenetic tree analysis showed that the gene was closely related to FBN1a and FBN1b in Arabidopsis thaliana. Homology analysis indicated that it was highly homologous to different FBN genes and its C-terminal part was particularly conserved in several kinds of plants. Quantitative analysis showed that NbFBN was highly expressed in leaves and flowers of Nicotiana benthamiana, and that the gene was found to be induced by drought stress and hormone ABA, indicating that the gene might be involved in response to the abiotic stresses.

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Genetic diversity of “Small & Medium Leaf” tea resources in Sichuan Province based on phenotypic characteristics

XIE Wengang, LI Xiaosong, LI Wei, TANG Qian

2019, 31(9): 1405-1415. DOI: 10.3969/j.issn.1004-1524.2019.09.02

Abstract ( 1103 )

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In order to protect and identify tea resources, and excavate special resources in Sichuan Province, principal component analysis (PCA) and cluster analysis were carried out on 30 phenotypic traits of 109 “ Small & Medium Leaf” tea resources in Sichuan. The results showed that the variation coefficient and diversity index of “Small & Medium Leaf” tea resources in Sichuan Province were 32.27% and 0.98, respectively. There were obvious differences in phenotypic traits of tea resources and rich diversity. Principal component analysis (PCA) and cluster analysis showed that the cumulative contribution rate of 12 principal component factors was 71.14%. The materials which were tested were clustered into 8 groups, and had little correlation with their provenance. The “Small & Medium Leaf” tea resources in Sichuan Province had rich phenotypic-traits genetic diversity, and phenotypic traits of different-provenance teas were different, which provided references for structure of core germplasm bank of Sichuan tea plants and selection of tea varieties.

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Association between the polymorphism of PvuⅡ locus of GH gene and growth traits in sheep

BAI Junyan, CAO Heng, WANG Xu, YANG Youbing, FAN Hongdeng, FU Xueyan, SHI Kunpeng, DONG Zhihao, LU Xiaoning, LI Xinyue, HAO Weiguang, LI Ziheng, ZHENG Feiyang

2019, 31(9): 1416-1422. DOI: 10.3969/j.issn.1004-1524.2019.09.03

Abstract ( 849 )

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To investigate the relationship between GH gene and growth and development performance of sheep, the polymorphism of PvuⅡ locus in intron Ⅱ of GH gene in five sheep populations was analyzed by PCR-RFLP, and the association between the polymorphism of PvuⅡ locus and sheep weight and body size was analyzed. The results showed that two genotypes, AB genotype (264 bp/429 bp/693 bp) and BB genotype (264 bp/429 bp), were detected at PvuⅡ locus of intron Ⅱ of GH gene in five sheep. The AB genotype frequencies of Large-tailed Han sheep, Small-tailed Han sheep, Yuxi fat-tailed sheep, Hu sheep and Dorper sheep were 0.786, 0.750, 0.424, 0.471 and 0.459 respectively, and BB genotype frequencies were 0.214, 0.250, 0.576, 0.529 and 0.541, respectively. The PvuⅡ locus of GH gene intron Ⅱ in Hu sheep deviated significantly from Hardy-Weinbery equilibrium (P<0.05), and the PvuⅡ locus of GH gene intron Ⅱ in Small-tailed han sheep, Yuxi fat-tailed sheep and Dupo sheep deviated significantly from Hardy-Weinbery equilibrium (P<0.01). The correlation analysis showed that the body weight, body length, chest width, buttock height, cannon circumference, rump height and neck length of AB genotype sheep were significantly higher than those of BB genotype sheep (P<0.05). The results showed that the PvuⅡ locus of intron Ⅱ of GH gene had some effects on the growth and development performance of sheep.

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Epidemiological investigation of porcine epidemic diarrhea virus and sequence analysis of ORF3 and S1 genes in Sichuan from 2016 to 2018

DEXI Cuomu, WANG Yin, YANG Zexiao, YAO Xueping, LUO Yan, LIAO Changyu, ZHANG Pengfei, JIANG Dike, XIANG Mingyuan, JIANG Ruijiao, SONG Yong

2019, 31(9): 1423-1428. DOI: 10.3969/j.issn.1004-1524.2019.09.04

Abstract ( 735 )

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To analyze the prevalence and molecular genetic evolution of pig epidemic diarrhea in Sichuan, this study conducted RT-PCR and related epidemiological investigations on 71 cases of diarrhea collected from June 2016 to June 2018, and ORF3 and S1 genes of four positive samples were tested and sequenced. The results showed that the peak incidence days in the past two years were January 28 and February 11, respectively, and the peak incidence period was from December to March. The number of infections in pig farms decreased year by year, and the new epidemic source increased. ORF3 gene can be divided into two genotypes, some strains had a small number of base mutations. The results of S1 gene high-variation segmentation showed that the prevalent PEDV strain from 2016 to 2018 was the GII group, which was the same as the current popular PEDV variant strain. The homology was 92.8%~99.3% and the number of base variation was large. The relationship with the domestic classic vaccine strain CV777 was far and the homology was 82.7%~87.3%. The incidence period was more concentrated than the traditional PEDV. The results showed that the incidence of PED in large-scale pig farms in Sichuan showed a downward trend, and preliminary results were obtained for the prevention and control of the disease. The current trend of PEDV strains was mutated. This study could lay the foundation for the prevention and control of PEDV.

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Isolation and identification of 4 strains of porcine pseudorabies virus and analysis of main virulence genes

YI Keke, YIN Wenqi, ZHOU Yuancheng, JIANG Jinzhen, ZHANG Baiyu, LI Zhongyin, YAN Qigui

2019, 31(9): 1429-1436. DOI: 10.3969/j.issn.1004-1524.2019.09.05

Abstract ( 947 )

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Pseudorabies virus(PRV)’s prevalence brings huge economic losses to the domestic pig industry, however, many large-scale immunized pig herds broke out pseudorabies, and gE antybodies turned from negative to positive since 2011, causing PRV clinical symptoms infected pigs, and then resulting in “abortion storm”. Most scholars believed that national outbreak of PRV might be related to factors such as viral virulence and genetic variation. In this study, four pseudorabies viruses were successfully isolated from suspected PRV infected pigs by PK-15 cells, moreover, their TCID₅₀ and LD₅₀ were determined. Their main immunogenic genes and virulence-related genes gB, gC, gE and TK were amplified, sequenced and analyzed. The 4 pseudorabies viruses were affirmed, named as FJ01, FJ03, YK and MS2018 strain, respectively, whose titers were 10^-6.63, 10^-7.08, 10^-8.10, 10^-7.18 TCID_50s·0.1mL^-1, respectively, and the results of virulence test in Balb/c mice showed as 10^2.17, 10^2.72, 10^3.44, 10^3.51 TCID_50s, respectively. The virulence of FJ01 strain was the strongest. Their nucleotide and amino acid homologies were analyzed and evolutionary trees were established. The results showed that the genes of FJ01, FJ03 and MS2018 strains had higher homology with the mutants such as HNX, HNB and JS-2012 strains which were isolated in recent years. In evolutionary relationship, they belonged to a large branch, however, they belonged to different evolutionary branches from vaccine Bartha-K61,SA215, and foreign strains Becker and Kaplan with evident variations. YK had the closer affinity with foreign strains in evolutionary relationship, but the reason remained to be further explored.

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Microbial community structure and diversity of mastitis cows by 16S rRNA high-throughput sequencing

ZENG Xueqin, LIU Chenjian, YANG Xue, LI Xiaoran

2019, 31(9): 1437-1445. DOI: 10.3969/j.issn.1004-1524.2019.09.06

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To clarify the pathogens that cause mastitis in cows, the V4-V5 hypervariable region of bacterial 16S rRNA gene in nipple wipes and milk of mastitis cows and normal cows were amplified and high throughput sequenced. The succession of microbial diversity was analyzed by biostatistics. The results showed that there was a very obvious difference in the composition of microbial between the nipple and the milk of the mastitis cow. At the phylum level, the highest abundance was Firmicutes in the nipples, while Proteobacteria was the highest in the milk. All nipples contain Bacteroidetes, while there is almost no in the milk, Fusobacteria only exists in nipples. The microbial community of the nipple and milk after mastitis in cows was different from normal cows. Fusobacteria was only detected on the nipples of mastitis cows, and normal cows were not. The abundance of Staphylococcus on the nipples of mastitis cows was higher than that of normal cows. Enterococcus was detected in mastitis cow milk, but not in normal cows. Therefore, the nipple of the cow was very different from the microbial community in the milk. Microbial composition changes both outside and inside the breast after bovine mastitis. Exogenous pathogens and endogenous sources pathogens can cause mastitis.

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Effects of DMSO and DMF on myocardial cytotoxicity and STAT3 signaling pathway in H9C2

YANG Min, ZHANG Kai, LI Jianxi, WANG Lei, ZHANG Kang, QIU Zhengying, WANG Xuezhi

2019, 31(9): 1446-1452. DOI: 10.3969/j.issn.1004-1524.2019.09.07

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In order to screen the lowest concentration of toxic effects of organic solvents DMSO and DMF on H9C2 cardiomyocytes, and to investigate the effects of DMSO and DMF on signal transduction and transcription factor 3 (signal transducer and activator of transcription 3, STAT3) signal pathway, the survival rates of cardiomyocytes exposed to DMSO and DMF at volume fraction of 0.1%, 0.2%, 0.3%, 0.5%, 0.7%, 0.9%, 1%, 2%, 3% for 24 h, 48 h and 72 h, respectively, were detected by the tetrazolium salt colorimetric assay (MTT method). And the survival rate of STAT3 and Bcl-2 protein was determined by Western blot method, and the expression of STAT3 protein was compared between DMSO and DMF at 0%, 0.2%, 0.3%, 0.5% volume fraction, respectively. At the same time, the survival rate of cardiomyocytes decreased with the increase of the volume percentage of DMSO and DMF, and the content of Bcl-2 and STAT3 protein decreased. When the volume fraction of DMSO was 0.3% and 0.5%, the cell survival rate was significantly lower than that of the control group (P<0.05). The protein levels of STAT3 and Bcl-2 were lower than those of the control group (P<0.05). The survival rate of cardiomyocytes decreased with the increase of the volume percentage of DMSO and DMF, and the content of Bcl-2 and STAT3 protein decreased at the same time. However, when the volume fraction of DMSO was 0.2%, there was no significant difference in the cell survival rate and the expression of STAT3 and Bcl-2 compared with the control group (P>0.05). When the volume fraction of DMF was 0.3% and 0.5%, the cell survival rate was statistically significant (P<0.05), and the protein levels of STAT3 and Bcl-2 were lower than those in the control group. When the volume fraction of DMF was 0.2%, the cell survival rate was not statistically significant (P>0.05). The expression of STAT3 protein was significantly higher than that in the control group (P<0.01). Compared with the control group (without DMSO and DMF), DMSO and DMF had toxic effects on H9C2 cardiomyocytes. With the increase of concentration, the cell survival rate decreased and the content of Bcl-2 protein decreased, which may be related to the down-regulation of STAT3 protein. The results of MTT assay were not completely consistent with those of Western blot test, and MTT assay was unstable.

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Prokaryotic expression and immunofluorescence localization of nucleoside diphosphate kinase gene from Dirofilaria immitis

ZHANG Haojie, LIU Mei, LI Chunyan, HE Ran, LAN Jingchao, LUO Li, GU Xiaobin, XIE Yue, YANG Guangyou

2019, 31(9): 1453-1460. DOI: 10.3969/j.issn.1004-1524.2019.09.08

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In order to provide base of heartworm diagnose, this study analyzed the molecular characteristics and identified the diagnostic potential of Dirofilaria immitis nucleoside diphosphate kinase (NDPK) gene. Di-NDPK was cloned and expressed, and the protein was analyzed by bioinformatic characterization, Western blotting and immunohistochemical analysis. Also the diagnostic potential of rDi-NDPK was evaluated by indirect ELISA. The results of Western blotting indicated that Di-NDPK had good reactivity. Immunohistochemical analysis results showed that Di-NDPK was distributed in hypodermis and intestinal epithelial cells of D.immitis. The indirect ELISA results showed that the sensitivity and specificity were 66.7% (16/24) and 38.9% (14/36) respectively. The cross reactions were observed with serum samples positive for Echinococcus granulosus, Ancylostoma caninum and Ascaris alata. In conclusion, Di-NDPK might be a candidate vaccine antigen, but not a suitable antigen for the detection of D. immitis.

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Cloning and bioinformatics analysis of GH gene and its flanking region in Odontobutis potamophila

LIU Jialin, LIU Shili, JIANG Wenping, CHENG Shun, CHI Meili, ZHENG Jianbo, JIA Yongyi, ZHAO Jinliang, YIN Shaowu, GU Zhimin

2019, 31(9): 1461-1470. DOI: 10.3969/j.issn.1004-1524.2019.09.09

Abstract ( 803 )

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The full-length sequence of the Odontobutis potamophila GH gene was successfully cloned by using long segment PCR, genome walking method and T-A cloning technology, which was submitted to GenBank to obtain the login number MH717101. The full-length of the GH gene of Odontobutis potamophila is 5 120 bp, the length of the 5'-end and 3'-end lateral sequence is 920 bp and 682 bp. It is predicted that the upstream region contains MF3, GAGA factor, hnf-3alpha, C/EBP, R2, GATA-1 and other transcriptional regulatory elements, respectively. The transcription unit is 3 518 bp long and contains 4 introns and 5 exons. 4 introns are 75, 333, 2 070 and 121 bp; 5 exons are 150, 131, 117, 138 and 373 bp, respectively. The third intron is 2 070 bp, the length was more than 4 times of the culter third intron. The open reading frame area (ORF) is 594 bp, encoding a protein polypeptide composed of 197 amino acid residues. Comparison of amino acid sequence homology between Odontobutis potamophila and Perciformes, Scorpaeniformes, Cypriniformes, Siluriformes with 13 fishes showed that the homology was the highest to Leucopsarion petersii, at 83.8%, and the homology to Ctenopharyngodon idella was the lowest, at 49.1%, the Odontobutis potamophila to show highly nonconservative at 180-197 amino acid sequence and the amino acid arrangement was completely different with the listed fish. The results of this study laid a molecular foundation for further study on expression, function and transcriptional regulation of GH gene in Odontobutis potamophila.

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Comparative study of several DNA extraction methods for raw animal muscle tissues

JI Yi, JIANG Yuanyuan, WANG Xiaofu, XU Xiaoli, XU Junfeng, LI Yueying, Chen Xiaoyun

2019, 31(9): 1471-1477. DOI: 10.3969/j.issn.1004-1524.2019.09.10

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DNA extraction methods with different principles for raw animal muscle tissue were conducted and compared to define the most suitable extracting method for the daily work of detecting as well as monitoring the adulteration in meat and meat products. The eight DNA extracting methods with various principles included the traditional CTAB-, proteinase K-, SDS-and rapid extraction-based methods, and the DNA extraction were conducted on raw beef, pork, lamb, chicken and duck. Moreover, the extracted DNA was evaluated for the integrity, yield, purity, time-consuming and inhibition effects on the real-time fluorescence quantitative PCR. As a result, all eight methods could be utilized to achieve the DNA extraction from the muscle tissues of common varieties of meat. Generally, all the commercial kits that based on the proteinase K, SDS or rapid extraction principle avoided organic solvents and exhibited a time-saving character when compared with the traditional CTAB-based method. Moreover, regarding to the higher DNA yield and purity as well as the less time-consuming and little fluorescence inhibition in real-time PCR, it gave priority to choose the commercial kits with proteinase K-or SDS-based method to extract DNA. Accordingly, these commercial kits could meet the requirement of the DNA extraction for detecting or monitoring the adulteration in raw muscle tissues of common varieties of meat.

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Cloning and expression of three chalcone synthase-like genes from Paeonia delavayi

YUAN Xiaolong, LI Yunqin, WANG Yi

2019, 31(9): 1478-1484. DOI: 10.3969/j.issn.1004-1524.2019.09.11

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Three CHS genes were isolated from the transcriptome of wild Paeonia delavayi with yellow flower, gene functions were predicted by using bioinformatics analysis, and gene expression levels in different developmental period of flowers were also analyzed. The results showed that the full-length cDNA of these three CHS genes were 1 173, 1 185, 1128 bp, respectively. They were named as PdCHS1 (GenBank ID was MK516264), PdCHS2 (GenBank ID was MK516265) and PdCHS3 (GenBank ID was MK516266), and encoded 390, 394 and 375 aa, respectively. The proteins encoded by these three CHS genes were non-secreted proteins and had no signal peptide, they all contained highly conserved catalytic active motif (G/A) FGPG. Phylogenic analysis indicated that these 3 proteins belonged to different branches. Expression level of PdCHS1 was high in the stamina and budding stage, expression level of PdCHS2 was high in the blossom falling stage and initial flowering stage, and expression level of PdCHS3 was high in the blossom falling stage and budding stage. These three CHS genes could participate in the different biosynthesis of secondary metabolites, PdCHS1 might relate to the naringenin biosynthesis, PdCHS2 might take part in the stibenoid biosynthesis, and PdCHS3 might involve in the polyketide biosynthesis.

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Effects of different cultivation patterns on soil microbial community and enzyme activity in continuous cropped pepper field

DONG Yufei, LYU Xiangzhang, ZHANG Zikun, HE Hongjun, YU Jingquan, ZHOU Yanhong

2019, 31(9): 1485-1492. DOI: 10.3969/j.issn.1004-1524.2019.09.12

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The effects of rotation and intercropping on soil microbial community and enzyme activity were studied by taking three years of continuous cropping of pepper, garlic-pepper rotation and maize-pepper intercropping rhizosphere soil as objects, which was expected to provide theoretical basis for relieving pepper continuous cropping obstacle. Results showed that rotation and intercropping significantly enhanced the activities of catalase and urease, and increased the biomass of bacteria and actinomycetes while the biomass of fungi was greatly decreased compared with continuous cropped soil (P<0.05). According to high-throughput sequencing results, the soil microbial structure of rotation field was similar to intercropping soil but far different from continuous cropped soil. Findings revealed that Proteobacteria and Acidobacteria were dominant bacterial populations, while Ascomycota and Basidiomycota were leading fungi phylum among the tested soil. Moreover, the relative abundances of Acidobacteria, Bacteroidete, Planctomycetes, Xanthomonada and Fusarium, Dothideomycetes, Zygomycota among different treatments were much significant (P<0.05). In conclusion, rotation and intercropping enhanced soil enzyme activity and soil bacterial community diversity. Besides, they improved soil microflora by reducing the relative abundance of potential pathogens while increasing the abundance of potential beneficial bacteria.

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Changes of physiological indexes and quality analysis of lettuce under optical interference film

ZHAI Keqing, ZHANG Xinyu, ZHOU Niannian, AN Yulan, LEI Yue, GAO Jie, GAN Defang, LIU Wen, HE Xiaomei

2019, 31(9): 1493-1501. DOI: 10.3969/j.issn.1004-1524.2019.09.13

Abstract ( 725 )

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Due to short growing period, high yield and suitable for raw food, lettuce becomes an important leafy vegetable in facility cultivation. Traditional photovoltaic power generation occupies a large amount of land resources, while photovoltaic agriculture performs photovoltaic power generation under the illumination demand that meets the growth of crops. In this study, two kinds of lettuce varieties, MonDai and Kristin were used as the tested varieties to study the bioactive enzymes, nutritional quality and the ultrastructure of the leaves under different optical interference films (red blue light transmission film and far red light-cutting film). The results showed that the CAT activity, SOD activity and malondialdehyde content of MonDai showed a downward trend under the red blue light transmission film and the far-red light-cutting film. Meanwhile, the increased activity of CAT and the reduced activity of SOD and the malondialdehyde appeared in Kristin covered with two films. However, the enzyme activities of two lettuces under two kinds of film mulching were mostly higher than those of the control. As for the nutritional quality of lettuce, the content of soluble sugar in the two lettuces increased firstly and then decreased in the red-blue light transmission film treatment, while there is an upward trend in the far red light-cutting film treatment. With the extension of the coverage time, the accumulation of vitamin C increased significantly in both light treatments and the content of protein and chlorophyll decreased, while most of them are higher than those of the control. The ultrastructural analysis of the leaves showed that the chloroplast aggregation of the MonDai increased under the red and blue light, while some cells of the leaves were destroyed under the far red light-cutting film, and the chloroplast granules were not obvious. This study provided a reference and basis for the technical improvement of optical interferometric film, the field application and the future development trend of photovoltaic agriculture.

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Study on preservation and germination of Cymbidium hybridum pollen

ZHANG Xiaoying, FU Qiaojuan, ZHAO Fukang, LI Chunnan, RUAN Ruoxin

2019, 31(9): 1502-1508. DOI: 10.3969/j.issn.1004-1524.2019.09.14

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To solve the problem of the flowering worst of orchid and provide reference for orchid hybridization breeding. This study have used the Cymbidium hybridum species JOIC and KK560 as test materials to screen the optimal medium for in vitro pollen germination, and detected the in vitro germination rate of Cymbidium hybridum pollen with different storage temperature and storage time to determine the appropriate pollen preservation method. The results showed that the optimum medium for in vitro germination of JOIC pollen was 0.5 MS+30% sucrose, and the optimum medium for in vitro germination of KK560 pollen was 0.01% H₃BO₃+0.2% CaCl₂+30% sucrose. Pollinium should be dried in the shade with silicone for 24 h before preservation and be preserved in -20 ℃ for six months.

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Karyotype analysis of 3 diploid hyacinth cultivars

SU Xiaoqian, WANG Fei, HU Fengrong

2019, 31(9): 1509-1515. DOI: 10.3969/j.issn.1004-1524.2019.09.15

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The karyotypes of three diploid hyacinth cultivars Gipsy Queen, Purple Sensation and Odysseus were analyzed by routine root tip pressing method to study the evolutionary degree and genetic relationship among different hyacinth cultivars. The results showed that karyotypes of hyacinths of three varieties were comparatively consistent, consisting of centromere chromosomes in the middle (m) and near-middle (sm), with a pair of satellites, but the distribution of satellites were differences in these three varieties. The karyotype formula of Gipsy Queen was 2n=2x=16=6m+8sm+2sm (sat), Purple Sensation was 2n=2x=16=6m+2m(sat)+8sm, and Odysseus was 2n=2x=16=6m+2m (sat)+8sm. The karyotype asymmetry coefficients of these three cultivars were 59.24%, 58.92% and 59.18%, respectively, and the karyotype types were all 2B.

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Research on automatic identification and counting of insect pests on sticky board

BAO Xiaomin, SHENG Jiawen

2019, 31(9): 1516-1522. DOI: 10.3969/j.issn.1004-1524.2019.09.16

Abstract ( 637 )

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In order to solve the arduous and inaccurate problem of manual counting of insect pests in yellow sticky board, based on the adaptive removal of the sticky board background and fusion of color space moments and image geometry, the quantification of pest characteristics were realized, and identification was carried out by weighted decision. Thus, an automatic identification and counting system was constructed. Taking Corcyra cephalonica on yellow sticky board as test materials, it was shown that the relative error between the results of the proposed system and manual counting on sticky board was less than 7%, indicating the feasibility of the proposed system.

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Prediction of soil available nitrogen content based on visible and near infrared spectroscopy preprocess and modeling

FANG Xiang, JIN Xiu, ZHU Juanjuan, LI Shaowen

2019, 31(9): 1523-1530. DOI: 10.3969/j.issn.1004-1524.2019.09.17

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In the present study, 188 yellow-red loam soil samples were collected in Southern Anhui, and the original spectrum was obtained by non-imaging spectrometer. Firstly, the characteristics of the average spectral reflectance curve at 350-1 657 nm were analyzed after preprocessing. Based on the original spectrum and spectra after 29 preprocesses, a total of 60 models were constructed either by partial least squares regression (PLSR) or radial basis function (RBF)-PLSR, and the constructed models were optimized and evaluated by the models' decision coefficient (R²) and relative percent deviation (RPD). It was shown that the PLSR model constructed on spectra after Savitaky-Golay filtering and log transformation (SG+LG/PLSR model) was most suitable for the prediction of soil available nitrogen content. Its R² and RPD were 0.94 and 3.88 in calibration set and were 0.91 and 3.38 in prediction set, which belonged to A level, and indicated that this model was feasible for soil available nitrogen content prediction.

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Evaluation of salt and alkali resistance of Pennisetum giganteum Z. X. Lin seedlings based on principal component analysis and membership function method

YAN Jingqiuzi, LI Gangtie, LIU Yujun, MA Yunxia, YANG Ying

2019, 31(9): 1531-1540. DOI: 10.3969/j.issn.1004-1524.2019.09.18

Abstract ( 946 )

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The plant height increase (H), water content (MC), relative water content (RWC), water saturation deficit (WSD), malondialdehyde content (MDA), catalase activity (CAT), superoxide dismutase activity (SOD), soluble sugar content (SS), free proline content (Pro), soluble protein content (SP), chlorophyll content (Chl) (including chlorophyll a content Chla, and chlorophyll b content Chlb) of Pennisetum giganteum Z. X. Lin seedlings were determined under simulated saline-alkali stress, and comprehensive evaluation of saline-alkali resistance was also carried out. It was shown that with the increase of saline-alkali concentration, H, RWC decreased, while the activities of CAT, SOD, and contents of SS, SP, Pro varied differently. When the concentration of test solution was 30 mmol·L^-1, and the ratio of NaCl, Na₂SO₄, NaHCO₃ concentration was 1:2:1, the comprehensive score of membership function was the highest (0.893), indicating good tolerance of Pennisetum giganteum Z. X. Lin under this simulated environment.

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Effect of aeration with carbon dioxide on growth of Desmodesmus sp. CHX1 in wastewater with high concentration of ammonium nitrogen

LUO Longzao, LIN Xiaoai, ZHU Feng, ZENG Fanjian, ZHANG Bangxi, LIU Ye, TIAN Guangming

2019, 31(9): 1541-1548. DOI: 10.3969/j.issn.1004-1524.2019.09.19

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The piggery wastewater can provide sufficient nutrient for the growth of microalgae. However, high concentration of ammonium nitrogen in the piggery wastewater may inhibit the growth of microalgae. In order to solve this problem, the inhibition of ammonia on Desmodesmus sp. CHX1 growth was investigated in the wastewater with initial ammonium concentration of 350-900 mg·L^-1, as well as the elimination effects of aeration with carbon dioxide. Results showed that the growth of Desmodesmus sp. CHX1 was inhibited by ammonia, with the increment of biomass of only 0.08-0.13 g·L^-1 after 7 days of culturing. Aeration with carbon dioxide could alleviate the inhibition of ammonia, with biomass increment of 0.32-0.47 g·L^-1. Decreasing trends of pH were observed in the wastewater without aeration, with the reduction of 0.5. While the pH in the aeration condition increased by 1.50 first and then decreased by 0.40. The ammonium was removed by 3.76%-20.10% in the experiment conditions. These results demonstrated that high concentration of ammonium nitrogen inhibited the growth of Desmodesmus sp. CHX1. Aeration with carbon dioxide could improve the growth of Desmodesmus sp. CHX1 in the wastewater with high concentration of ammonium nitrogen.

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Effect of apple polyphenols on quality and browning of fresh-cut lotus root

ZHANG Hanqing, LIU Ruiling, WU Weijie, DENG Shanggui, GAO Haiyan, CHEN Hangjun

2019, 31(9): 1549-1554. DOI: 10.3969/j.issn.1004-1524.2019.09.20

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Fresh-cut lotus roots were treated with different concentrations (0.1%, 0.3%, 0.5%) of apple polyphenols to study its effects on the browning and quality by monitoring the changes of color, crispness, ascorbic acid and total phenolis content, the activity of polyphenol oxidase (PPO) and peroxidase (POD) during storage. The results showed that apple polyphenol treatment significantly inhibited the browning of fresh-cut lotus roots. Compared with other treatment groups, 0.1% apple polyphenol treatment maintained better color and brittleness of lotus root, delayed the degradation of the ascorbic acid and total phenolic, slowed down the increase of malondialdehyde content and inhibited the activities of PPO and POD. The results showed that apple polyphenol treatment could maintain the quality of fresh-cut lotus root.

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Inhibition effect of Ageratina adenophorais methanol extracts against Microsporidium gypsum and its mechanism

HU Liwen, ZHANG Yong, SHI Zhen, FU Jie, SUN Wei, DENG Junliang, REN Zhihua, ZUO Zhicai, CAO Suizhong, HU Yanchun

2019, 31(9): 1555-1562. DOI: 10.3969/j.issn.1004-1524.2019.09.21

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In the present study, 9-oxo-10,11-dehydroagerophorone (euptox A), cadinan-3-ene-2,7-dione (CED) and olefin compounds were separated and identified from Ageratina adenophorais methanol extract by silica gel column chromatography, thin-layer-chromatography and gas chromatography-mass spectrometer. The inhibition effects of 125, 62.5, 31.25, 15.6, 7.8, 3.9, 1.95 and 0.98 μg·mL^-1 euptox A and CED against Microsporidium gypsum were determined at 48, 72, 96, 120, 144 h, respectively. The Microsporidium gypsum samples treated with euptox A were selected as test materials, dye penetration test with propidium iodide (PI) was carried out, and the content of total protein and total phosphorus in extra-cellular fluid, and ergo-sterol content in mycelium were determined. It was shown that the separated euptox A and CED from Ageratina adenophorais methanol extract could inhibit the growth of Microsporidium gypsum, and euptox A possessed stronger inhibition effect. After the treatment with euptox A, the hyphae of Microsporidium gypsum showed strong red fluorescence under fluorescence microscope. The contents of total protein and total phosphorus in extracellular fluid of Microsporidium gypsum were significantly (P<0.05) increased, yet the ergo-sterol content in mycelium was significantly (P<0.05) decreased. It was inferred that euptox A could inhibit the synthesis of ergo-sterol, cause membrane damage, and lead to cellular disorder and cell death.

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Evaluation and spatial correlation of cultivated land utilization efficiency under net carbon emission constraint of China

WU Haoyue, HUANG Hanjiao, HE Yanqiu, CHEN Wenkuan

2019, 31(9): 1563-1574. DOI: 10.3969/j.issn.1004-1524.2019.09.22

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In order to explore the effective utilization mode of cultivated land and grasp the cooperative mechanism under the constraint of environment, the dual effects of cultivated land utilization were taken into consideration, which consisted of carbon emission and absorption. The SBM-Undesirable model was adopted to evaluate the cultivated land utilization efficiency of 30 provinces (municipalities, autonomous regions) in China from 2007 to 2016. Besides, based on the matrix of spatial distance squares' reciprocal and Moran's I, the spatial agglomeration of the cultivated land utilization efficiency was discussed from both global and local perspectives. It was shown that the overall cultivated land utilization efficiency was relatively high, of which the average reached up to 0.867, yet different regions showed diverse performance and could be divided into 5 grades. Most of the high-efficiency areas were located in the southeastern coastal area, the western area and the northeastern area, while the low-efficiency areas were mainly situated in the northern coastal area and the middle reaches of the Yellow River. The majority of regions with low efficiency showed redundant inputs and net carbon emissions. There was a certain spatial agglomeration of the cultivated land utilization efficiency within provinces, and the spatial correlation pattern experienced a process of “multi-core distribution” to “one string and one area” to “low pole dominance”. Attentions should be paid to control the unexpected output and optimize the input, strengthen regional cooperation according to local conditions, and explore the path of efficient utilization of cultivated land with regional coordination.

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