Acta Agriculturae Zhejiangensis

Bioinformatics analysis of StUOXs gene family in potato

LIANG Liqin, YANG Rui, GAO Gang

2020, 32(9): 1523-1532. DOI: 10.3969/j.issn.1004-1524.2020.09.01

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Potato(Solanum tuberosum) is the third largest food crop in the world, and its yield and quality are adversely affected by biotic and abiotic stresses. Ubiquinone oxidase(UOX), as a mitochondrial terminal oxidase encoded by nuclear genes, plays an important role in the regulation of plant growth and stress response. In order to understand the response of StUOXs in potato disease resistance, five StUOXs genes were identified in potato genome by bioinformatics methods, and their physical and chemical properties, chromosome location, tertiary structure, evolutionary relationship, conserved domain, promoter element and expression profile, especially the expression patterns induced by Ralstonia solanacearum were analyzed. The results showed that the number of amino acids encoded by potato StUOXs genes was between 279 and 366, the molecular weight was distributed between 31 and 42 ku, and the subcellular locations were located in mitochondria. Phylogenetic analysis showed that potato StUOXs members were closely related to tomato and pepper UOXs members, but slightly farther than Arabidopsis thaliana and tobacco. FPKM data analysis showed that StUOXs were mainly induced by salt, hormone and biological stress in leaves, which corresponded to the cis-acting elements such as hormone response in its promoter. The qRT-PCR test after inoculation with Ralstonia solanacearum showed that StUOX4 was involved in the early response to the bacterial wilt pathogens.

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Transcriptome analysis of red bud taro and green stem taro in Yanshan, Jiangxi Province

YIN Minghua, CAO Qing, CHEN Hong, DENG Siyu, DENG Yanmei

2020, 32(9): 1533-1543. DOI: 10.3969/j.issn.1004-1524.2020.09.02

Abstract ( 829 )

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In order to explore the molecular differences between Jiangxi Yanshan red bud taro and Jiangxi Yanshan green stem taro, the transcriptome analysis was carried out on the plantlets of Jiangxi Yanshan red bud taro (HYY group) and Jiangxi Yanshan green stem taro (QGY group). The results showed that the Clean reads of HYY group was 41 298 676, and the GC content was 53.09%; the Clean reads of QGY group was 45 551 860, and the GC content was 51.17%. In HYY group and QGY group, the pair value of expression FPKM was between 0-2, and the expression density was between 0-0.7. The number of common genes expressed in HYY group and QGY group was 14 038, the number of genes independently expressed in HYY group was 17 843, the number of genes independently expressed in QGY group was 18 634. The correlation coefficient of the expression of HYY group and QGY group was 0.035, and the correlation between samples was poor. There were 32 555 differential expressed genes (DEG) in HYY group and QGY group. Compared with QGY group, there were 15 887 up-regulated genes and 16 668 down regulated genes in HYY group. Go enrichment analysis showed that differential genes were mainly annotated to polysaccharide metabolic process, gene silencing, pectin catabolic process, carbohydrate metabolic process, auxin-activated signaling pathway, extracellular region, intrinsic component of membrane, integral component of membrane, membrane part, apoplast, hydrolase activity, acting on glycosyl bonds, hydrolase activity, hydrolyzing O-glycosyl compounds, protein dimerization activity, DNA binding, oxidoreductase activity, acting on diphenols and related substances as donors and other functions. KEGG enrichment analysis showed that the differential genes were mainly annotated to plant hormone signal transduction, plant-pathogen interaction, cysteine and methionine metabolism, phenylpropanoid biosynthesis, cortisol synthesis and secretion, peroxisome, MAPK signaling pathway-plant, mismatch repair, starch and sucrose metabolism, sulfur metabolism, pentose and glucuronate interconversions, C-type lectin receptor signaling pathway, glycerolipid metabolism, biosynthesis of unsaturated fatty acids, alpha-linolenic acid metabolism, diterpenoid biosynthesis, flavonoid biosynthesis, fatty acid elongation, photosynthesis, indole alkaloid biosynthesis and other pathways. The results of this study will provide reference for variety identification and molecular breeding of Jiangxi red bud taro and Jiangxi green stem taro.

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Comparative analysis on genetic diversity of Colocasia esculenta germplasm in Zhejiang Province based on phenotype and simple sequence repeats markers

HONG Xia, ZHAO Yongbin, QU Weidong, CHEN Yinlong, QIU Liping, WANG Jiaoyang

2020, 32(9): 1544-1554. DOI: 10.3969/j.issn.1004-1524.2020.09.03

Abstract ( 915 )

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In order to explore the genetic diversity of Colocasia esculenta germplasm in Zhejiang Province, and excavate special resources, 25 landraces were selected in the experiment. Genetic diversity analysis, principal component analysis (PCA), and cluster analysis were carried out on 15 phenotypic traits, and simple sequence repeats (SSR) molecular marker cluster analysis was combined based on these 25 resources. The results showed that the variation coefficient and diversity index were 31.32% and 0.97, respectively, indicating Colocasia esculenta germplasm in Zhejiang Province had a rich genetic diversity. Principal component analysis (PCA) showed that the accumulated eigenvalue of 5 principal components was 10.871, and cumulative contribution rate was 77.649%. The first principal component factor reflected the plant morphology of multi-cormels taro, the second principal component factor reflected the shape of taro leaf, the third principal component factor reflected the shape of parent-taro, the fourth and fifth principal component factor represent parent-taro epidermis hair (PTEH) and parent-taro colour, respectively. Moreover, grandson-taro shape (GTS), parent-taro bud colouring (PTBC), leaf junction colour (LJC), lower middle petiole colour (LMPC) and parent-taro epidermis hair (PTEH) could be preferentially observed in the Colocasia esculenta germplasm identification. All resources could be clustered into five categories at the genetic distance of 4.171 based on phenotypic traits. Among them the shape of son and grand-son taros in group Ⅲ and group Ⅴ was oval, which showed a well commodity nature. Otherwise, two special taro germplasm were excavated with purplish red petiole. 20 pairs of primers amplified 53 bands, of which 48 bands had polymorphism. The polymorphism rate was 90.6%. Polymorphism information content (PIC) ranged from 0.22 to 0.64. 25 taro accessions could be grouped into 5 clusters at DICE genetic similarity coefficient 0.721 based on SSR markers, this was similar to the phenotypic clustering, but also had some difference. Two clustering methods were supplement of each other among Colocasia esculenta germplasm identification, indicating there was no direct relationship between geographical origin. These results would provide an important theoretical basis for the further conservation, utilization and innovation of Colocasia esculenta germplasm.

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Analysis of nicotine content and related gene expression in different types of tobacco germplasm

XU Xiuhong, LIU Jinliang, LI Dongcheng, LIU Renxiang

2020, 32(9): 1555-1563. DOI: 10.3969/j.issn.1004-1524.2020.09.04

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In order to study the changes of nicotine content and expression of related genes in different types of tobacco germplasm, four tobacco types, including flue-cured tobacco K326, oriental tobacco Basma, burley tobacco TN90 and sun-cured tobacco Maryland were used as materials. Nicotine content in different leaves and the expression levels of nicotine metabolism related genes PMT, QPT, CYP82E5V2 and A622 were measured at different developmental stages. The results showed that with the passing of growth stage, the nicotine content increased slowly, and increased dramatically after topping. Except for Basma, the nicotine content continued to increase after topping, while in K326, TN90 and Maryland, the nicotine content increased sharply after topping, then decreased slowly, and gradually stabilized. PMT gene in different types of tobacco was expressed in different growth stages. Expression level of PMT was higher at the seedling stage, and its change trend was basically the same as that of the nicotine content. Expression level of QPT was higher in the early stage, then decreased, and reached a peak one week before topping. After topping, the expression level continued to increase, which was positively correlated with the change of nicotine content. Expression level of CYP82E5V2 remained at a high level in different growth stages, presenting a wave pattern, and increased in the bloom stage, and peaked before and after topping. However, expression level of CYP82E5V2 in Basma decreased when it was mature. Expression level of CYP82E5V2 was positively correlated with the change of nicotine content before the topping, and negatively correlated to a certain extent from the later topping stage to the mature stage. Expression level of A622 in K326 and Maryland rose in a wave-like manner, and remained at a high level at maturity, while in TN90 and Basma, expression level of A622 was higher one week before and after topping, then decreased, and increased again, the expression level was lower at maturity, and there was no obvious consistency with the change of nicotine content.

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Tissue expression and significant target genes analysis of swine miR-204

WANG Wei, GUN Shuangbao, WANG Pengfei, HUANG Xiaoyu, XIE Kaihui, LUO Ruirui, GAO Xiaoli, ZHANG Bo, YAN Zunqiang, YANG Qiaoli, MA Yanping

2020, 32(9): 1564-1573. DOI: 10.3969/j.issn.1004-1524.2020.09.05

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Based on the earlier research, miR-204 was screened out with significantly up-regulated expression in the resistance and susceptibility group for piglets infected with Clostridium perfringens type C by using Illumina Hiseq 4000 high-throughput sequencing technology. In the present study, real-time quantitative PCR was used to detect the miR-204 expression in different tissues of piglets at 7-day-old. Meanwhile the mature gene sequences was analyzed using MEGA7.0 software, and target genes were predicted using TargetScan, miRDB, and PicTar. Then, the online software DAVID was used to perform gene ontology and KEGG pathway enrichment analysis of target genes, and qRT-PCR method was used to verify the targeted relationship of the selected target genes in porcine intestinal epithelial cells (IPEC-J2). The results showed that miR-204 was highly expressed in kidney, liver, thymus, lymph, duodenum and ileum tissues of 7-day-old piglets, and its mature gene sequences was highly conserved among vertebrates. There were 114 common target genes predicted by the three softwares, and these target genes were significantly (P<0.05) enriched in 15 GO functions and 13 signaling pathways. Combined with the previously obtained resistance genes, the four key target genes nuclear receptor subfamily 3 group C member 1(NR3C1), Silence information regulator 1 (SIRT1), BCL2 like 2(BCL2L2) and Discs, large homolog 5(DLG5) were screened. After transfection of miR-204 mimics, the target genes SIRT1, BCL2L2 and DLG5 were significantly (P<0.01)down-regulated. In summary, miR-204 is an important factor involved in regulating the piglets' resistance caused by Clostridium perfringens type C infection. The genes SIRT1, BCL2L2, and DLG5 may be key target genes of miR-204, and the further study should be conducted to testify their functions.

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Effect of variation of PRKCB gene intron 14 in Sansui ducks on eggshell quality

LIAO Chaomei, YOU Minfang, TAN Guanghui, LI Jiezhang, ZHANG Yiyu, QIN Yuanyu, LIU Ruoyu

2020, 32(9): 1574-1580. DOI: 10.3969/j.issn.1004-1524.2020.09.06

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To explore the genetic effect of PRKCB gene on eggshell quality of Sansui duck, direct sequencing of PCR products, MegaAlign program of DNAStar software and Chromas software were used to screen and identify the genetic locus variation of (Anas platyrhyncha domestica) PRKCB gene. The correlation between SNP loci and eggshell traits were analysed by SPSS 18.0 software. The results indicated that three moderately polymorphic novel SNPs of g.1158465 C>T, g.1158519 G>A and g.1158524 G>A were detected in intron 14 of PRKCB gene in Sansui duck. Three genotypes were produced in each SNP. The genotype distribution of g.1158519 G>A and g.1158524 G>A were in accordance with Hardy-Weinberg equilibrium, while that of g.1158465 C>T was significantly (P<0.05) deviated from Hardy-Weinberg equilibrium. There was a strong linkage disequilibrium between g.1158519 G>A and g.1158465 C>T loci. The combination of three SNPs composed of 5 haplotypes and 8 diplotypes. The frequency of haplotype H5 (TGG) and diplotype H2H5 was 0.492 and 0.200, respectively. Association analysis showed that the eggshell thickness of individuals with AG genotype at g.1158519 G>A locus was significantly (P<0.05) higher than that of individuals with GG genotype, and the eggshell thickness of diplotype H1H3 individuals was significantly (P<0.05) higher than that of H3H5 individuals. The results suggested that the genetic variation of the intron 14 in PRKCB gene can affect duck eggshell quality, and g.1158519 G>A locus may be used as a genetic marker to improve duck eggshell quality.

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Effect of temperature on physiology metabolism of Meretrix petechialis

HU Gaoyu, ZHANG Xiang, LIN Xingguan, CAI Yilong, CAI Jingbo, XIAO Guoqiang, CHAI Xueliang

2020, 32(9): 1581-1590. DOI: 10.3969/j.issn.1004-1524.2020.09.07

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In the present study, Meretrix petechialis was selected as the study object. Based on the laboratory still water method and intermittent respiration measurement method, the effects of different temperature on ingestion rate, oxygen consumption rate, faecal egestion rate, and ammonia excretion rate of Meretrix petechialis were explored. It was shown that the maximum values of ingestion rate, ammonia excretion rate, faecal egestion rate and oxygen consumption rate were found under 25-30, 20-30, 15-20, and 35 ℃, respectively. With the elevated water temperature, the ingestion rate and ammonia excretion rate of Meretrix petechialis increased first and then decreased, while the faecal egestion rate gradually decreased, yet oxygen consumption rate increased. Under the experiment conditions, the variation ranges of ingestion rate, ammonia excretion rate, faecal egestion rate and oxygen consumption rate were 0.015-0.043 mg·g^-1·h^-1, 1.26-135.25 μg·g^-1·h^-1, 2.173-15.908 μg·g^-1·h^-1 and 0.290-1.780 mg·g^-1·h^-1, respectively. These results could provide references for the study of physiological metabolism of Meretrix petechialis in different seasons and guide the development of corresponding mariculture.

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Complete sequence and gene organization of mitochondrial genome of Megalobrama terminalis from Qiantang River

LIU Kai, FENG Xiaoyu, MA Hengjia, XIE Nan

2020, 32(9): 1591-1608. DOI: 10.3969/j.issn.1004-1524.2020.09.08

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To study the mitochondrial genomic structure of Megalobrama terminalis Richardson from Qiantang River and phylogenetic relationship of Cultrinae subfamily, the amplification by PCR, sequencing, and software splicing were used to obtain complete mitochondrial genome of M. terminalis from Qiantang River, whose GenBank accession number was MN725725. The full-length sequence of the mitochondrial genome of M. terminalis from Qiantang River was 16 621 bp, and its base composition was A (31.23%), G (16.17%), C (27.87%), and T (24.73%). There were 13 protein-coding genes, 22 tRNA genes, 2 rRNA genes in the mitochondrial genome. Except for NAD6, tRNA-Gln, tRNA-Ala, tRNA-Asn, tRNA-Cys, tRNA-Tyr, tRNA-Ser^(UCN), tRNA-Glu, and tRNA-Pro, the other genes were encoded on the H chain. The A+T content of the mitochondrial genome and protein-coding genes in the mitochondria of M. terminalis from Qiangjiang River was 55.97% and 55.86%, respectively. The mitochondrial genome, and protein-coding genes in the mitochondria had obvious AT preference. There were two interspersed repeats in the mitochondrial gene, which were located at the front end of the terminal associated sequences region and the 3'end of the control region in the mitochondrial control region, respectively. Of the 22 tRNA genes, except tRNA-Ser^(AGY), they had a typical clover secondary structure. Based on the comparison of BLAST, the consistency of M. terminalis from Qiantang River and Heilongjiang River was 99.76%, while the consistency of M. terminalis from Pearl River was 99.87%. Phylogenetic tree was constructed based on complete mitochondrial genomes of 15 species of Cultrinae subfamily belonging to 7 genera. The genetic relationship between Megalobrama and Parabramis was closer than that of Culter, and Megalobrama was far away from Hemiculter, Hemiculterella, and Metzia.

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Effect of chicken-derived compound probiotics on immunoglobulin and toll-like receptor pathway of white feather broilers

GUI Xueer, WANG Zhi, LI Siting, HE Mengchu, ZHU Jie, FENG Shibin, WU Jinjie

2020, 32(9): 1609-1614. DOI: 10.3969/j.issn.1004-1524.2020.09.09

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To investigate the effects of probiotics on serum immunoglobulin and intestinal Toll-like receptor pathway in White feather broiler, one hundred and eighty 28-day-old white feather hybrid broilers were randomly divided into group I (basic diet), group II (basic diet adding 10¹¹ CFU·kg^-1 probiotics) and group III ((basic die adding 2×10¹¹ CFU·kg^-1 probiotics), with 3 repeats in each group and 20 broilers in each repeat. The experimental period was 21 d. The results showed that, compared with group A, the serum IgG and IgM levels in group B and group C increased significantly (P<0.05). The protein expression of TLR2, TLR4, Myd88, TRAF-6, AP-1 and their relative expression of mRNA in group B and group C were significantly (P<0.05) increased. Compared with group B, the serum IgG, IgM and IgA levels in group C were no significant difference (P>0.05). The protein expression of TLR4 and MyD88 in group C were significantly (P<0.05) increased, and the relative expression of MyD88 mRNA was significantly (P<0.05) increased. Feeding probiotics can improve the serum IgG and IgM levels of white feather broilers, and regulate the immune function of broiler intestine effectively through toll like receptor pathway.

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Analysis of simple sequence repeats in transcriptome of garlic (Allium sativum L.) and development of molecular markers

LIU Xinyu, TIAN Jie

2020, 32(9): 1615-1625. DOI: 10.3969/j.issn.1004-1524.2020.09.10

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In order to investigate features of SSR sequence and develop SSR primers of Allium sativum for garlic breeding, MISA software was used to detect the SSR loci and analyse the SSR information of the garlic transcriptome sequence in this study. The effectiveness and polymorphism of primers were determined by PCR amplification. The results showed that 444 865 unigenes were obtained from garlic transcriptome sequencing. A total of 141 132 SSR loci were identified, the occurrence frequency was 31.72%, the average distribution distance was 3.45 kb. The major repeat motifs were mononucleotide and dinucleotide, which accounted for 68.02% and 24.12%, respectively. 82 repeat motifs of SSR loci were identified, and the most abundant motifs were A/T and AT/AT,which accounted for 65.02% and 12.37%, respectively. A total of 125 616 pairs of SSR primers were designed by Primer 3.0, and 14 pairs of primers were randomly selected and amplified. There were 12 pairs of primers that were clearly amplified. Among them, 6 pairs showed rich polymorphism in 35 garlic germplasm. 26 polymorphic bands were obtained by using 6 pairs polymorphic primers, and the average polymorphic amplification band of each SSR primer was 4.33. According to the UPGMA analysis, 35 garlic germplasm resources were divided into 5 groups at the genetic similarity coefficient of 0.756 9. In conclusion, abundant SSR loci were obtained by development of SSR molecular markers in garlic transcriptome and SSR markers could be widely used.

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Effects of different mulching methods on yield, quality and soil nutrients of open-air loose-curd cauliflower on plateau in summer

LI Jinwu, YU Jihua, LYU Jian, FENG Zhi, YANG Haixing, CHE Xusheng, QIN Qijie, ZHANG Yang, JIN Ning

2020, 32(9): 1626-1633. DOI: 10.3969/j.issn.1004-1524.2020.09.11

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In order to find out the effect of different mulching methods on yield, quality and soil nutrients of the open ground loose-curd cauliflower on theplateau in summer, the field experiment, five mulching methods were adopted, including plastic film and straw mulching (T1), straw mulching between rows (T2), whole straw mulching (T3), open field without mulching (CK1) and plastic film mulching (CK2), the yield, quality, mineral element content, irrigation amount, irrigation utilization efficiency, and soil nutrient content of loose-curd cauliflower were measured and analyzed. The results showed that the surface mulching could reduce the irrigation amount in the whole growth period of the loose-curd cauliflower, and the yield and water production efficiency of T1 treatment were higher than those of other treatments. Compared with CK1 and CK2, T1 could save water by 29.0% and 7.3%, increase production by 68.9% and 4.7%, and irrigation water use efficiency increasedby 6.12 and 1.22 percent points. The content of soluble sugar and vitamin C in T1 treatment was higher than that in other treatments, and the content of nitrate was the lowest. Compared with CK1 and CK2, T1, T2 and T3 treatment could improve the mineral element content of flower ball, and T3 treatment had the most significant effect. The content of soil organic matter, total phosphorus in T1 treatment was 29.6%, 108.4% higher than that in CK1, and the content of total nitrogen, total potassium, soil calcium, magnesium and manganese had no significant difference from CK1; The content of soil organic matter, total nitrogen, total phosphorus and manganese in T1 treatment was 14.7%, 80.0%, 52.0% and 105.3% higher than that in CK2, and the content of total potassium, soil calcium and magnesium had no significant difference with CK2. The treatment of plastic film and strawmulching had the effect of conserving soil moisture and increasing production, improving the quality of loose-curd cauliflower and improving the soil environment, effectively realizing the recycling of crop straw, avoiding the environmental problems caused by local straw burning, and promoting the sustainable development of agriculture in this area.

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Effects of salinity level on photosynthetic characteristics of Sargassum fusiforme seedlings

SONG Xindan, CHEN Binbin, MA Zengling, XU Lili, LIN Lidong, WU Mingjiang

2020, 32(9): 1634-1644. DOI: 10.3969/j.issn.1004-1524.2020.09.12

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Sargassum fusiforme seedlings were used to determine their photosynthetic responses to different seawater salinities (15‰, 20‰, 29‰, 45‰). The results showed that photosynthesis of S. fusiforme was significantly affected by excessive low-and high-salinity stress. Specifically, at salinity of 20‰, photosynthesis of S. fusiforme seedling was slightly affected, which indicated low salinity tolerance in S. fusiforme seedlings. However, excessive low salinity level (15‰) significantly depressed algal photosynthesis. At high salinity of 45‰, S. fusiforme seedlings were damaged by high salt stress, its photosynthetic activity and utilization of inorganic carbon were significantly inhibited. In a short time treatment(1-12 h), seedling photosynthesis was slightly affected, and it's pigment contents, maximum relative election transport rate (rETR_max), and maximal photochemical efficiency (F_v/F_m) exhibited changes of decreasing followed by increase. In prolonged culture(240 h), excessive low-(15‰) or high-salinity (45‰) significantly inhibited algal photosynthesis, meaning notable decreasing of its rETR_max, light use efficiency(α), maximum net photosynthesis rate (P_nmax) and photosynthetic carbon fixation (v_max). Moreover, excessive high salinity stressed the photosynthesis process of S. fusiforme seedling by hindering the synthesis of its photosynthetic pigments. It indicated that the strategies for preventing low-and high-salinity stress on S. fusiforme seedlings should be practically performed when the salinity level extremely changed, to avoid the severe stress of prolonged salinity change on the seedlings.

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Transcriptome analysis on development of aerial root in grape of Weike

GE Jintao, WANG Jiangying, ZHAO Wenjing, SHAO Xiaobin, ZHU Pengbo, TANG Xueyan, SUN Mingwei, LIU Xingman

2020, 32(9): 1645-1655. DOI: 10.3969/j.issn.1004-1524.2020.09.13

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Aerial root is a special type of root in grape which grows under high temperature and high humidity. The aerial root has low developmental degree and high potential for differentiation. In this study, techniques were used to induce aerial germination and rooting at the stem nodes of the fruiting branch of Weike grape, and the expression of root transcription genes was analyzed with the untreated stem nodes of the same mother branch as the control. The results showed that the expression of 3 989 genes was increased and the expression of 2 830 genes was down-regulated during the induction of aerial roots. GO functional classification was used to classify annotated genes into 40 functional categories, including catalytic activity, binding, and metabolic processes. Parts of the root key gene expression results showed that all plant hormones including auxin, gibberellin and abscisic acid were involved in the regulation process. Combined with corresponding gene analysis of auxin downstream, endoglucanase, PRE gene and high-affinity nitrate transporter gene significantly up-regulated in the aerial root development, indicating that the degradation of epidermal cell wall at stem nodes and the enrichment of nitrate at stem nodes to stimulate root germination were important regulatory mechanisms for the growth and development of aerial roots.

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Demonstration on rice pests control by fertilizer regulation technique in different geographic rice growing areas of Zhejiang Province

ZHENG Xusong, ZHONG Liequan, WANG Huifu, CHEN Fangjing, CHENG Liping, XU Qiqiang, LI Yang, ZHONG Xuhua, LYU Zhongxian

2020, 32(9): 1656-1664. DOI: 10.3969/j.issn.1004-1524.2020.09.14

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The practice of fertilizer regulation technique (FRT) for rice pests control in single cropping rice season was carried out in different geographic rice growing areas of Zhejiang Province, including paddy fields in Yuyao (plain), Linhai (hill), Longquan (basinin mountain area) and Jingning (terrace), with rice varieties of Zhejing 88, Yongyou 18, Yongyou 1540 and Zhongzheyou 2838 planting, respectively. FRT guideline included reducing nitrogen, applying less nitrogen in early season, increasing potassium and optimizing the ratio of nitrogen, phosphorus and potassium, and its effects on rice agronomic characters and yield, and main diseases and insect pests were compared with those using local farmers' traditional fertilization. The results showed that FRT could obviously reduce the occurrence of rice main diseases and insect pests. On average, the planthopper density, leaf folded rate by leaffolder, and rates of rice false smut and sheath blight were decreased by 64.79%,30.57%,14.76% and 64.82%, respectively. In Yuyao, Linhai and Longquan, average rice yields were increased by 5.88%, even though the average nitrogen and phosphorus fertilizer input were reduced by 19.31% and 31.80%, respectively. Meanwhile, the partial factor productivity from applied nitrogen (PFPN) were increased by 50.26%, 28.93% and 44.29%, respectively. In the terrace area of Jingning, FRT led to slight yield loss, and the PFPN was decreased by 2.62%. However, when basic seedlings were increased by 20% under FRT, the rice yields were increased by 16.67% and the PFPN was increased by 18.05%. All these results indicated that FRT could not only reduce nitrogen application, reduce the occurrence of major diseases and pests in rice, but also increase production. In conclusion, FRT was applicable in all rice planting areas in Zhejiang Province.

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Passivation effects of calcium carbonate with/without chitosan on chromium pollution in calcareous soil

CHANG Huiqing, XU Fujin, PAN Yajie

2020, 32(9): 1665-1671. DOI: 10.3969/j.issn.1004-1524.2020.09.15

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In order to explore the effective method for chromium (Cr) remediation in calcareous soil, a field experiment was carried out to study the passivation effect of calcium carbonate with/without chitosan on Cr pollution. It was shown that in the test soil, the main form of chromium was residual, which accounted for 71.28% of the total Cr content. The addition of exogenous Cr significantly (P<0.05) increased the content of various forms of Cr in soil, especially the oxidizable Cr, which accounted for more than 76% of the total Cr. While, the content of water-soluble Cr was the smallest, and the smallest number was only 0.45%. Addition of calcium carbonate with/without chitosan significantly (P<0.05) reduced the content of water-soluble Cr and significantly (P<0.05) increased the content of residual Cr. Cr-contaminated soils were passivated by addition of calcium carbonate with/without chitosan, of which the passivation rate was increased by 35.44 and 21.64 percent, respectively. The contents of malondialdehyde (MDA) and soluble protein, and catalase (CAT) activity in maize leaves were increased under Cr pollution. MDA content in maize leaves was significantly (P<0.05) decreased by 31.92% and 20.57% by addition of calcium carbonate with/without chitosan. Compared with addition of calcium carbonate alone, the addition of calcium carbonate with chitosan significantly (P<0.05) reduced Cr content in maize grains, roots and stems, and significantly (P<0.05) reduced the enrichment factor of Cr in maize grains. The present study provided method reference for the remediation of Cr contaminated calcareous soil.

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Growth and arsenic enrichment characteristics of Typha angustifolia L. under different arsenic pollution levels

CHEN Tian, BAO Ningying, DU Chongxuan, LIU Yungen

2020, 32(9): 1672-1682. DOI: 10.3969/j.issn.1004-1524.2020.09.16

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To investigate the growth status of typical wetland emergent plant Typha angustifolia L. and its characteristics of arsenic (As) accumulation and migration, As₀, As₅₀, As₁₅₀ and As₆₀₀ (corresponding to 0, 50, 150 and 600 mg·kg^-1 As of sediment) were set up to study the growth and development, As accumulation characteristics of T. angustifolia L. during the life cycle (growth period, maturity period and yellow wilting period) and the effect of As extraction from soil. The result showed that T. angustifolia L. could complete its life cycle and formed seed stigma under four levels of As stress. In the growth stage, the root tolerance index (IT) of T. angustifolia L. under As₅₀ treatment was the highest, reached to 132.38%, and the biomass of T. angustifolia L. under As₁₅₀ treatment was 19.12 g, 34.08% higher than that under As₀ treatment, the biomass accumulation under As₆₀₀ treatment was 12.35 g, which was 13.39% less than that under As₀ treatment, at this time, typha IT was the smallest, which was 65.23%; In the mature stage, the biomass accumulation of typha under As₅₀ treatment was the highest, reaching 72.23 g, 28.70% higher than that under As₀ treatment, at this time, IT of typha was the highest, reached to 149.25%; The biomass accumulation and IT under As₆₀₀ treatment were the lowest. In the yellow wilting stage, the biomass accumulation and IT of typha under As₅₀ treatment were the highest, reached to 79.23 g and 119.28%, respectively; The biomass accumulation under As₆₀₀ treatment was the lowest, 13.47% lower than that under As₀ treatment. In the three stages, GSH activity of typha was the highest when As content was 0-50 mg·kg^-1 in the sediment, and F_v/F_m in As₅₀ and As₁₅₀were higher than that in As₀; F_v/F_m in A₆₀₀ was the lowest, and SOD activity and MDA content were the highest in the leaves; The bioaccumulation coefficient (BCF) of As in the underground part of typha was higher than that in the aboveground part, the transfer coefficient (TF) between plants was 0.09-1.10. The As fixed rate (ER) in the sediment of typha was higher than the removal rate (RR), and there was a significant positive correlation between the amount of extraction (EA), the amount of migration per unit area (M_PUA) and the amount of As in the sediment. Typha culd endure high concentration As stress and grow normally, and had good remediation effect on the sediment of As polluted wetland. It could be planted as a wetland plant establishment species in As polluted wetland, and could increase the number of harvest and replacement of typha before the yellow wilting stage to obtain better remediation effect.

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Screening and utilization of ammonia-nitrogen-degrading microorganism

SUN Xiaojun, SHEN Qi, WU Yifei, YAO Xiaohong, LI Yuancheng, SUN Hong, WANG Xin, TANG Jiangwu, GE Xiangyang

2020, 32(9): 1683-1691. DOI: 10.3969/j.issn.1004-1524.2020.09.17

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To obtain the ammonia-lowering agent for livestock manure, the bacterial strains with good ammonia-nitrogen-degrading capability were isolated from swine manure. Three strains with high ammonia-nitrogen degradation efficiency were identified with the olfactory threshold method combined with the comparisons of their ammonia-nitrogen degradation efficiencies, and were encoded as Z2, AOZ and Y, respectively. Based on the results of morphological observation, physiological and biochemical tests, and 16S rRNA sequence homology analysis, the strains Z2, AOZ and Y were identified as Bacillus flexus, Pseudomonas putida, and Lactobacillus casei, respectively. The experimental results showed that the ammonia-nitrogen degradation efficiency was higher when strains Z2 and Y were mixed, and the highest ammonia-nitrogen degradation efficiency reached 53.19%, suggesting that the microorganism complex could be practically applied to remove ammonium from swine manure.

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Dynamic analysis of river nitrogen and phosphorus pollution based on LOADEST model and wavelet transform

JIN Yanan, ZHANG Baifa, HAO Yun, WU Jianhong, LYU Jun

2020, 32(9): 1692-1701. DOI: 10.3969/j.issn.1004-1524.2020.09.18

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The estimation of pollutant loads in rivers and the analysis of their variations are the basis for non-point source pollution treatment. However, the low-frequency manual monitoring cannot reflect the daily continuous changes of water quality in rivers. Based on the monthly water quality monitoring data at the outlet section of Changle River in Zhejiang Province from 2003 to 2016, the LOADEST model for total nitrogen (TN) and total phosphorus (TP) load estimation in the river was calibrated and verified. Then, the daily TN and TP loads in the Changle River were estimated by using the continuous flow data. The results showed that, during 2003-2016, the average annual TN and TP loads in the river were (2 207.71±862.48) t·a^-1 and (71.43±31.56) t·a^-1, respectively, while their daily average loads were (6.04±11.81) t·d^-1 and (0.19±0.26) t·d^-1, respectively. Both the TN and TP loads had an increasing trend in the study period. Meanwhile, the variation patterns of TN and TP loads in the river during the research period were analyzed by using wavelet analysis method. It was found that the TN load in the Changle River had periodic changes with a main time span of 18 months. The variation of the TP load was similar to that of the TN load, but the varying intensity was relatively weak. The wavelet variance of TN and TP loads suggested that the periodic variations of TN and TP loads were mainly dependent on the periodic changes of river flow, yet less affected by TN and TP concentrations.

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Design and experiment on washer with two-speed hair brush rollers for fruits

HE Xiaodong, ZHU Dequan, ZHU Jianjun, LIAO Juan, ZHANG Shun, LI Jingfa

2020, 32(9): 1702-1710. DOI: 10.3969/j.issn.1004-1524.2020.09.19

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In order to solve the problems of low cleaning rate and high damage rate of spherical crispy fruits during the cleaning process, the differential speed principle of brush roller was adopted to design a washer with dual-speed brush rollers for fruits which had the functions of soaking, brushing, spraying and collecting fruit. Its basic structure and working principle were explained, and its key components such as soaking device, spraying device, brushing device and so on were designed and analyzed. Red Fuji apple was selected as the test material. The rotation speed of the odd-numbered brush rollers, the rotation speed of the even-numbered brush rollers, and soaking time were taken as test factors. The cleaning rate and damage rate were used as evaluated indicators. The single factor test was executed to determine the levels of factors. Through the orthogonal test with three-factor and three-level, the significance of the influence of the test factors on each test index was analyzed, and the optimal cleaning parameter combination for fruit cleaning was determined. The results showed that the rotation speed of the odd-numbered rows of brush rollers was 80 r·min^-1, the rotation speed of the even-numbered brush roller was 100 r·min^-1, and the soaking time was 25 s. Under these conditions, the fruit cleaning rate was 98.08%, and the damage rate was 3.07%.

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Study on price relevance of livestock products based on VEC-BEKK-GARCH and spillover index model

LIU Jiaojiao, ZHANG Gangren, GAO Qun

2020, 32(9): 1711-1721. DOI: 10.3969/j.issn.1004-1524.2020.09.20

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With monthly price data of three animal products pork, beef and mutton from January 2001 to March 2019, the spillover effect within animal products price was studied by using VEC-BEKK-GARCH(1,1) model and spillover index. It was shown that there was a significant (P<0.05) two-way mean spillover effect between the market of pork and beef, as well as the market of beef and mutton, while there was only one-way mean spillover effect from the pork market to the mutton market. The volatility spillover effect within the price of livestock products was obvious, which contributed 38% to the total forecast error variance, and the spillover effect lasted six months. In the short term, the spillover effect of pork price shocks on beef and mutton price shocks was relatively weak, only 6.8% and 0.94% respectively. In the long term, the spillover effect was enhanced, and the total spillover effect on beef and mutton price shocks reached 39.9%. At the same time, the shock of pork prices was also affected by other price, up to 19.1%. The two-way volatility spillover effect between beef and mutton prices was obvious, about 27.5%. The spillover effect between the price of livestock products posed a challenge to the regulation of livestock products prices. Therefore, the spillover direction and extent between livestock products prices should be fully considered to improve the early warning mechanism of livestock products prices.

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Support level and structural characteristics of maize subsidy policy in world's major maize producing countries and region

GUO Yanjing, XIAO Haifeng

2020, 32(9): 1722-1731. DOI: 10.3969/j.issn.1004-1524.2020.09.21

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Based on a review of the development of maize subsidy policy in the United States (US), the European Union (EU), Brazil and China, the policy evaluation system of Organization for Economic Co-operation and Development (OECD) was used to compare and analyze the support level and structural changes of maize subsidy policies in the above countries and region from 1995 to 2018. In terms of support level of maize subsidy policy, the EU has the lowest level, and is followed by Brazil and the US. While China's policy support level for maize is relatively high. China should coordinate subsidy funds, appropriately adjust the amount of subsidies for maize, and actively guide farmers to gradually adjust the planting structure. In terms of support structure for maize subsidy policy, the policy support in the US and EU for maize is mainly payments based on current area (PC), and it is mainly market price support (MPS) in Brazil. As affected by border trade protection measures, such as maize tariff quota management, policy support for maize in China is still dominated by MPS, which has caused some distortions in the market. China should continue to push forward the reform of the maize storage system to make the maize price formed by the market, and gradually increase the international competitiveness of China's maize.

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Research progress of mustard in Zhejiang Province

MENG Qiufeng, HU Meihua, WANG Jie, REN Xiliang, GAO Tianyi, CHEN Jianming

2020, 32(9): 1732-1740. DOI: 10.3969/j.issn.1004-1524.2020.09.22

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Mustard industry is relatively developed, and there are a large variety of products for us. Zhejiang Province is one of the main producing areas of mustard in China. More than 10 sub-species of mustard are grown in Zhejiang. Brassica juncea var. tumida and Brassica juncea var. crispifolia are the most important two kinds of mustard with long planting history in Zhejiang Province. Processing products of pickled tuber mustard and potherb mustard are famous in the world. It has attracted the attention of vegetable research scholars for a long time and achieved certain research progress. Therefore, this paper will review the research progress of the basic research, genetic breeding and processing technology of mustard in Zhejiang province, and the problems in the mustard study were summarized and the future research directions were prosposed in order to provide reference for the follow-up research on mustard.

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