Acta Agriculturae Zhejiangensis

CRISPR/Cas9 editing and expression analysis of LOC_Os10g05490 in rice under cold stress

SHEN Chunxiu

2017, 29(2): 177-185. DOI: 10.3969/j.issn.1004-1524.2017.02.01

Abstract ( 849 )

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To explore the expression and cold tolerance function of LOC_Os10g05490 in rice under cold stress, semi-quantitative analysis was performed for Dongxiang wild rice and 93-11. The results showed that expression level of LOC_Os10g05490 in 93-11 was the same before and after cold stress, but it was down regulated in Dongxiang wild rice after cold stress. This was consistent with RNA-seq data and further study indicated that this gene might be associated with cold tolerance in Dongxiang wild rice. Plant expression vectors with CRISPR/Cas9 genome editing technique were subsequently constructed to target the LOC_Os10g05490 locus, and were transformed into the japonica rice variety TP309. A total of 26 transgenic positive plants were obtained. This study may lay the foundation for subsequent screening of mutant plants and identification of genes associated with cold tolerance in rice.

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Sequence analysis of ABA-insensitive gene of common buckwheat, Fagopyrum esculentum

WANG Yan, LIU Yao, LIANG Chenggang, CHEN Qingqing, SHI Taoxiong, CHEN Qijiao, MENG Ziye, CHEN Qingfu

2017, 29(2): 186-192. DOI: 10.3969/j.issn.1004-1524.2017.02.02

Abstract ( 609 )

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ABA-insensitive (ABI) is a kind of transcription factors that regulates plant stress response, seed shattering and dormancy. Bioinformatic analysis indicated that the cDNA sequence of common buckwheat ABI(CL38501) gene contained 963 bp, coded 320 amino acids including the bZIP domain and the conserved domain Ⅱ of ABI5, suggesting that it might encode the ABI5 family protein. The major genes involve in ABA signal transduction pathway were up-regulated in cultivar including ABI that was up-regulated by 4.42-fold and 3.66-fold at bud stage and flowering stage, respectively, as compared with the wild germplasm of common buckwheat. A total of 405 sites in ABI gene including 30 polymorphic sites were identified among 9 cultivars and 5 wild germplasms of common buckwheat. Only 1 polymorphic site was identified among 9 cultivars, whereas 7 polymorphic sites were identified in 5 wild germplasms. These results indicated that the ABI sequence of common buckwheat was highly conserved. A total of 11 consistent polymorphic amino acid sites were identified among cultivar and wild germplasm. The change of protein conformation was found between cultivar and wild germplasm of common buckwheat induced by the amino acid transformation at the 32nd and 60th sites of ABI, implying that it might be associated with the loss of seed shattering and dormancy during the long domestication process.

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Isolation, identification and phylogenetic analysis of Lactobacillus from vagina of giant panda

MA Xiaoping, YANG Tianyi, YU Yan, ZHANG Zhihe, WANG Chengdong, GU Yu

2017, 29(2): 193-199. DOI: 10.3969/j.issn.1004-1524.2017.02.03

Abstract ( 532 )

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In order to separate and identify the Lactobacillus from vagina of giant panda, the vaginal secretions were taken from estrus female giant panda and women. The isolated Lactobacillus strains were identified by morphology, biochemical characteristics, 16S rDNA sequencing and phylogenetic analysis. The results showed that a total of 6 gram-positive Lactobacillus strains were obtained. Lactobacillus salivarius GPV01, Lactobacillus plantarum GPV02, Lactobacillus saerimneri GPV03 were isolated from the vaginal secretion of the giant panda (test group), and Lactobacillus acidophilus WAV01, Lactobacillus crispatus WAV02, Lactobacillus plantarum WAV03 were isolated from the vaginal secretion of the woman (control group). It was interesting that the giant panda and woman all had L. plantarum in the vagina (GPV02 and WAV03). This result would lay the foundation for studying biological property of vagina Lactobacillus of giant panda and exploiting vaginal probiotics for giant panda.

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Effects of spermidine on immune organ index and expression of inflammatory cytokines in mice

XU Qilin, KANG Bo, JIANG Dongmei, WEI Chunye, JIA Yilin, YI Zhixin, CHEN Ziyu

2017, 29(2): 200-205. DOI: 10.3969/j.issn.1004-1524.2017.02.04

Abstract ( 445 )

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To clarify the effects of exogenous spermidine on mice immune organs index and expression of inflammatory cytokines in thymus and spleen, 0, 0.05, 0.10 and 0.15 mg·g^-1 (body weight) spermidine were intraperitoneal injected for the six-week Kunming mice. The thymus and spleen index were measured after 24 h and the expression of TNF-α、IL-1β、IL-6 and IFN-γ in thymus and spleen were assessed by quantitative real-time PCR. The results showed that, compared with the control group, intraperitoneal injection of 0.15 mg·g^-1 spermidine group significantly (P<0.05) decreased the thymus index and the spleen index. 0.1 mg·g^-1 spermidine group enhanced the thymus IFN-γ expression (P<0.05). In the spleen, the expressions of TNF-α and IL-1β in 0.05 mg·g^-1 spermidine group were significantly (P<0.05) lower than those of the control group, but the expressions of the IL-6 and IFN-γ gene in 0.1 mg·g^-1 spermidine group were significantly higher than those of the control group (P<0.05). The expression of TNF-α in 0.15 mg·g^-1 spermidine group was significantly (P<0.05) lower than that of the control group. Thus, the spermidine showed tissue specific and dose dependent manner on the expression of inflammatory cytokines in mice. Low concentration of spermidine had anti-inflammatory effect by inhibiting the expression of inflammatory cytokines, but high-dose spermidine could also induce inflammation.

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Effect of Wuling San on copper poisoning in experimental rats

CHENG Yan, LIU Yayun, ZHANG Fangjie, HUANG Cuihua, KANG Youxi, HAN Chunyang, CHEN Dingding, LIU Cuiyan

2017, 29(2): 206-212. DOI: 10.3969/j.issn.1004-1524.2017.02.05

Abstract ( 513 )

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In order to investigate the effects of Wuling San, a Chinese herbal medicine compound powder with promoting diuresis, on the copper excretion in animal, the pathological model of subchronic copper poisoning in SD rats was established by feeding high copper diet, and then Wuling San as a therapeutic drug was given by means of intragastric administration to rats. After the success of building models and at the end of treatment, the blood sample of rats was collected by cardiac puncture for determining copper content with the method of flame atomic absorption spectrometry, and the activities of serum AST, LDH and SOD were detected by reagent kit. At the end of the experiment, all the rats were anatomized to observe the changes of pathologic autopsy of the internal organs, and the histopathologic changes of liver and kidney. Results showed that compared with control group, the rats of the model group were slow in velocity of body weight increasement, the obvious pathological changes of liver and kidney structure were found and the level of blood copper was significantly higher (P<0.01), indicating that the model of subchronic copper poisoning in rats was successfully established. After treated with Wuling San, compared with model group, the treatment group had less detectable changes in histopathology, the level of blood copper was significantly lower (P<0.05), SOD activity increased (P<0.05) and AST and LDH activities declined (P<0.05). The above five indexes had no significant differences between the treatment group and the control group (P<0.05). The results of this study indicated that Wuling San could effectively promote the excretion of copper in rats, alleviate the damage of liver and kidney, so it could be used to treat clinical subchronic copper poisoning of animals.

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Structural analysis and identification of biological activity of alpha-toxin amino-terminal PLC from Clostridium welchii type A

XU Chongli, XU Chongbo, GONG Yuchen

2017, 29(2): 213-219. DOI: 10.3969/j.issn.1004-1524.2017.02.06

Abstract ( 511 )

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Amino-terminal PLC1-250 gene of Clostridium welchii α-toxin was amplified by PCR. The recombinant strain BL21 (DE3)(pN-PLC1-250) containing PLC1-250 gene was constructed. It was shown that the recombinant plasmid pN-PLC1-250 contained PLC1-250 gene with correct sequence and ORF by identification of endonuclease-digesting and sequence analysis. SDS-PAGE analysis showed that the expression level of PLC1-250 proteins were about 18.76% of total cellular proteins. The secondary structure and three-dimensional structure of PLC1-250 proteins were predicted by SOPMA method on EXPASY website. The results showed that the secondary structure of PLC1-250 protein was composed of alpha helices and random coils. The three-dimensional structure of PLC1-250 protein was similar with amino-terminal domain of α-toxin protein. The study of biological activity laid foundation for further research of the molecular mechanism of α-toxin and the relation of its molecular structure and biological function.

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Cloning and expression analysis of FabHLH3 gene from cultivated strawberry and construction of overexpression and silencing vector

ZHANG Qing, MIAO Lixiang, ZHANG Yuchao, YANG Xiaofang, JIANG Guihua

2017, 29(2): 220-227. DOI: 10.3969/j.issn.1004-1524.2017.02.07

Abstract ( 754 )

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The full-length cDNA sequence of FabHLH3 gene was cloned from strawberry (Fragaria×ananassa cv. Yuexin) through homology cloning method. Then, bioinformatics and expression pattern of FabHLH3 gene were analyzed by some softwares and real-time fluorescent quantitative PCR technique, respectively. Sequence analysis showed that the cDNA of FabHLH3 was 2 094 bp, encoding 697 amino acids, while the estimated molecular weight and isoelectric point of the putative protein were 77.43 ku and 6.05. After DNAMAN analysis, it shared 92.04% and 90.87% identity with the nucleotide sequence and amino acid sequence of bHLH3 gene from woodland strawberry(F. vesca), respectively. The expression of FabHLH3 was analyzed in leaf, petiole, flower and fruit being grown under colored light-quality plastic film. The results showed that FabHLH3 was expressed in all the tissues and the highest level was found in leaf. The expression analysis showed that the expression level of FabHLH3 was gradually increased during fruit development. The lowest expression was detected at small green fruit stage, and a reduced peak appeared at the full red stage. In addition, the expression of FabHLH3 gene can be induced by red and blue light. The overexpression and silencing vectors of FabHLH3, pGreen-FabHLH3 and pGreen-FabHLH3i, were constructed by in-fusion method with a plant expression vector pGreenII 62-SK. It would be used in functional analysis in the future research.

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Genetic diversity analysis of Brassica oleracea L. var. italica with SSR markers

ZHANG Zhixian, HE Daogen, ZHU Changzhi, TAN Guoyin, GAO Xu

2017, 29(2): 228-235. DOI: 10.3969/j.issn.1004-1524.2017.02.08

Abstract ( 618 )

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Simple sequence repeat (SSR) markers were used to analyze genetic diversity among 28 broccoli (Brassica oleracea L. var. italica) materials. Fourteen pairs of SSR marker primers selected from 50 pairs of SSR primers generated 94 polymorphic bands among these 28 materials. The polymorphism information content (PIC) ranged from 0.20 to 0.91. The overall genetic similarity values ranged from 0.585 1 to 0.904 3 with an average value of 0.728 9. Among them, TN12 and TN14 were the most similar while TN503 and TN597 were the most distant ones with genetic similarity values of 0.904 3 and 0.585 1, respectively. The cluster analysis divided the 28 broccoli materials into two clades (Ⅰand Ⅱ). Clade Ⅰ was constituted of 26 broccoli inbred lines and clade Ⅱ were formed by TN539 and TN597. While the result of principal coordinates analysis showed that all 28 materials were grouped into 4 clades, which consist of 2, 6, 6, 14 broccoli germplasm materials, respectively. Both of the two methods showed that TN539 and TN597 had a low genetic similarity with other 26 germplasm materials. These results revealed that 28 broccoli materials had a narrow genetic base while part of them had wide variations, which provide information for their use and the hybrid parents choice.

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Effects of NaCl treatments on growth and ion distribution of Portulaca oleracea seedlings

DONG Jing, XING Jincheng, HONG Lizhou, WANG Maowen, ZHU Xiaomei, LIU Chong, WEN Zhugui, ZHAO Baoquan, DING Hairong

2017, 29(2): 236-243. DOI: 10.3969/j.issn.1004-1524.2017.02.09

Abstract ( 435 )

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A sand culture pot experiment was conducted in order to investigate the effects of 3 NaCl stress levels of 0, 100, 200 mmol·L^-1 on seedling growth and distribution of Na⁺, K⁺, Ca²⁺ and Mg²⁺ in different organs of Portulaca oleracea. The results showed that under different levels of NaCl stress, dry weight of shoots and leaves number were decreased, while dry weight of roots and root/shoot ratio were increased. Among these three organs, content of Na⁺ was significantly increased compared with that of the contrast seedlings, and mainly concentrated in roots. Moreover, K⁺ contents dramatically decreased, and the decrease range of the K⁺ contents was larger in leaves and smaller in stems. Ca²⁺ and Mg²⁺ contents in leaves were dramatically higher than those in stems and roots. With the increase of NaCl stress concentration, ratios of K⁺/Na⁺, Ca²⁺/Na⁺, Mg²⁺/Na⁺ were decreased, primarily due to the increasing accumulation of Na⁺. Besides, ratios of K⁺/Na⁺, Ca²⁺/Na⁺, Mg²⁺/Na⁺ in leaves were relatively higher than those in stems and roots. The ability of ion-selective transportation from roots to stems of Portulaca oleracea seedlings increased first and then decreased, and that from stems to leaves gradually increased with the increasing NaCl stress. The results of comprehensive analysis indicated that Portulaca oleracea possibly protect seedlings from damage caused by NaCl stress via compartmentalization of Na⁺ into roots and increasing the upward transportation of K⁺, Ca²⁺ and Mg²⁺.

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Analysis of combining ability and heritability for main characters of melon fruit

SHEN Jia, SHOU Weisong, ZHANG Yuejian

2017, 29(2): 244-250. DOI: 10.3969/j.issn.1004-1524.2017.02.10

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Melon is one of the main fruits of the world, and the main objective for melon breeders is to breed new varieties with high quality. To improve the efficiency of melon hybrid combinations, the combining ability and heritability for mainly characters of melon fruit were analyzed by 28 cross combinations which were made with 8 melon inbred lines by the method of Griffing (IV). Except for vertical diameter of fruit, the other six characters of melon fruit including diameter of fruit heart chamber, horizontal diameter of fruit, vertical diameter of fruit, flesh thickness, soluble sugar at center and edge were significantly different between the inbred lines. And A1, A3 and A4 showed the high general combining ability (GCA) effect value in fruit weight, diameter of fruit heart chamber, horizontal diameter of fruit and flesh thickness, while A2, B1, B2, B3 and B4 showed the high GCA effect value in fruit sugar at flesh center and edge. A1×A2, A1×B3, A3×B2 and A3×B3 were the excellent hybrid combinations with high special combining ability (SCA). Our results provided reliable basis for selecting proper parental lines in cross breeding of melon.

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Suitability for processing green tea from high amino acid content of tea introduced to Hubei Province

MA Linlong, JIN Xiaofang, CAO Dan, LIU Yanli, GONG Ziming, WANG Shengpeng, LIU Panpan

2017, 29(2): 251-260. DOI: 10.3969/j.issn.1004-1524.2017.02.11

Abstract ( 686 )

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The suitability for processing green tea from three high amino acid of tea varieties (Huangjingcha 1, Anji Baicha and Zhonghuang 1) introduced to Hubei Province was investigated by comparison with Echa 1, which was the dominant cultivar in Hubei Province. One-bud-two-leaf green teas from three introduced cultivars were analyzed for major biochemical components, aroma components and sensory quality. The results showed that content of water extract, tea polyphenols, catechin, ratios of tea polyphenols and amino acids of these three kinds of green tea were all significantly lower than those of the control, but total amino acid and 18 kinds of amino acid were significantly higher than those of the control except leucine, phenylalanine. A total of 37, 35 and 36 aroma constituents were indentified in Huangjincha 1, Anji Baicha and Zhonghuang 1, respectively. The types, components and proportions of aroma constituents in these three tea varieties were different from the Echa1. The sensory quality detection showed that Anji Baicha was the most excellent one, followed with the Huangjincha 1, and Zhonghuang 1 showed a similar level with Echa1. Therefore, the green tea products, which made from the three cultivars introduced to Hubei Province, showed quality characteristics of high grade green tea with slight bitterness and astringency, rich aroma, refreshing and velvety taste.

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Effects of forestry ecological projects on landscape pattern evolution and migration of barycenter in Dengkou County

ZHANG Qibin, YUE Depeng, YU Qiang, LYU Qi, YIN Bo, MA Huan, LI Ning

2017, 29(2): 261-269. DOI: 10.3969/j.issn.1004-1524.2017.02.12

Abstract ( 510 )

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To explore the influence of forestry ecological project on the local environment, the landscape evolution characteristics of Dengkou County during 2000 and 2014 were studied using multifarious landscape analysis methods. On the basis, the landscape pattern of Dengkou County in 2021 was simulated using CA/Markov model under the following conditions: either with or without the landscape transfer probabilities caused by forestry ecological project. Then, the transfer characteristics of the barycenter of landscape types under each simulation condition were analyzed. It was shown that during the 14 years studied, the area of water, forestry land and construction land increased obviously, while the area of desert decreased apparently. Forestry land, desert, construction land and water had higher dynamic degrees. The landscape dynamic degrees in the border area of different landscape types were relatively high. The barycenter of water and forestry migrated to the south-west of the county and the barycenter of desert migrated back to the hinterland of the Ulanbuh Desert markedly. The simulation results with the landscape transfer probabilities caused by forestry ecological project indicated that the landscape pattern of Dengkou County would show an ecofriendly trend of evolution, while the landscape would deteriorate without the landscape transfer probabilities caused by forestry ecological project. The study results indicated that the forestry ecological project played key role in environment improvement in Dengkou County.

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Identification of twig blight pathogens of Myrica rubra and analysis on soil nutrient contents in Yunnan Province

WU Yangchun, REN Haiying, QI Xingjiang, ZHENG Xiliang, LIANG Senmiao

2017, 29(2): 270-276. DOI: 10.3969/j.issn.1004-1524.2017.02.13

Abstract ( 614 )

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In order to clarify the cause of bayberry twig blight in Yunnan Provinces pathogens were isolated and identified by Koch's rule, morphology and rDNA-ITS sequences. The soil nutrient contents of diseased planting-stocks, diseased container seedlings, healthy container seedlings and native healthy bayberry trees were tested. The results showed that the pathogens were identified as Pestalotiopsis versicolor. The contents of phosphorus, boron, potassium, calcium and zinc in diseased planting-stocks significantly increased with 55.8%-511.5% and nitrogen increased with 9.2%, compared with the healthy planting-stocks, which meaned that the planting-stocks grew in the rich-nutrient soil. In addition, compared with the natively healthy bayberry trees, phosphorus and potassium in the diseased container seedlings were significantly increased with 203.2% and 431.6%, respectively, magnesium increased with 27.6%, other nutrient contents were all decreased, with 28.6%-96.9%, which indicated that nutrient contents in container seedlings soil were imbalance. The results of this study provided important base for bayberry disease prevention and control in Yunnan Province.

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CgRGS4 regulates vegetative growth, osmotic pressure response, oxidative stress response and pathogenicity in Colletotrichum gloeosporioides

XU Shuang, WU Manli, KE Zhijian, LIU Zhiqiang, LI Xiaoyu

2017, 29(2): 277-285. DOI: 10.3969/j.issn.1004-1524.2017.02.14

Abstract ( 466 )

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Regulators of G-protein signaling (RGS) are a kind of negative regulatory factor of G protein, which play an important role in growth development and pathogenicity of plant pathogen. However, there has been no research on biological functions of RGS in Colletotrichum gloeosporioides up to now. The aim of this study was to clone the gene of CgRGS4 and analyze its biological function in C. gloeosporioides. The CgRGS4 was cloned by PCR, and its gene-knockout mutant was obtained by homologous recombination. The biological functions of CgRGS4 were preliminarily determined through phenotypic analysis. The results showed CgRGS4 encoded a 1 224-amino acids protein, containing RGS, PXA and PX function domains. Compared with the wild type, the knockout mutant of CgRGS4 had slow growth on poor medium, enhanced tolerance to osmotic stress, low melanin content, sensitivity to H₂O₂, decrease in extracellular laccase and catalase activity, decreased pathogenicity. In conclusion, CgRGS4 is involved in regulation of vegetative growth, osmotic pressure response, oxidative stress response and pathogenicity of C. gloeosporioides.

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PCR-DGGE assisted selection of ammonia degrading bacteria

ZHANG Zhichao, SUN Hong, WU Yifei, WANG Xin, YAO Xiaohong, LIU Yong, TANG Jiangwu, GE Xiangyang

2017, 29(2): 286-291. DOI: 10.3969/j.issn.1004-1524.2017.02.15

Abstract ( 534 )

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In the present study, strains able to efficiently degrade ammonia in livestock waste water were screened with traditional enrichment selection method combined with PCR-denaturing gradient gel electrophoresis (DGGE). Twenty-two strains were screened out after domestication under different ammonia concentrations. Among them, 3 strains exhibited relatively higher ammonia degradation abilities, and were named as ZZC-3, ZZC-4 and ZZC-14, respectively, which were identified as Gordonia sp., Denitrobacter sp. and Rhodococcus sp. by 16S rDNA sequence analysis. The PCR-DGGE results confirmed that the Denitrobacter sp. had the highest concentrations in the enriched solution of waste water. Therefore, strain ZZC-4 was selected as the target strain. After incubation for 144 h, the treatment with strain ZZC-4 reached the highest degradation efficiency, which was 90.8% for ammonia nitrogen and 94.7% for COD, respectively.

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Effects of different materials on base cations leaching in island greening soils

CHEN Wen, WU Haiping, WANG Jing, MA Wanzhu

2017, 29(2): 292-299. DOI: 10.3969/j.issn.1004-1524.2017.02.16

Abstract ( 476 )

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In order to study the effects of organic fertilizer, wood chips, biochar and gypsum on base cations (K⁺, Ca²⁺, Na⁺, Mg²⁺)leaching, soil column simulation experiment was carried out. It was shown that addition of any above material could effectively increase the leaching amount of base cations, and improve soil quality. Under the experiment conditions, addition of 25 g organic fertilizer or gypsum, respectively, led to the highest leaching amount of K⁺, Ca²⁺, Na⁺, Mg²⁺ as compared to other doses. As for wood chips, the leaching amount of Ca²⁺, Na⁺, Mg²⁺ reached the highest level with addition of 12.5 g wood chips, while the leaching amount of K⁺ was the highest without wood chips. As for biochar, the leaching amount of Ca²⁺, Mg²⁺ reached the highest level with addition of 12.5 g biochar, while the leaching amount of K⁺, Na⁺ was the highest with addition of 25 g biochar. Comprehensively, the following solutions were effective for base cations leaching: (1) Irrigation. This method was simple and time-saving, but, it cost a lot of water. (2) Adding amendment. Based on the findings in the present study, addition of 25 g organic fertilizer, 12.5 g wood chips, 12.5 g gypsum exhibited the best leaching effect and the highest efficiency under the set conditions.

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Review on application of willows (Salix spp.) in remediation of contaminated environment

ZHAO Fengliang, YANG Weidong

2017, 29(2): 300-306. DOI: 10.3969/j.issn.1004-1524.2017.02.17

Abstract ( 593 )

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Willows had ideal biological characteristics for phytoremediation, such as high biomass, fast-growing, easy propagation and high transpiration ability. Relevant studies showed that willows could effectively remove nitrogen, phosphorus from contaminated environment through plant-soil, plant-sand, constructed wetland and river buffer zones. Willows could remediate organic pollutants such as ethanol, polychlorinated biphenyls and polycyclic aromatic hydrocarbons. Willows could also remove heavy metals through natural phytoremediation and induced phytoremediation (chelators, arbuscular myconhizal fungi). The phytoremediation efficiency of willows was strongly correlated with species or clones, soil types and hydrological regime. In the future, more strength should be put toward to explore the bioenergy potential of willow along with environment remediation techniques, and more efforts should be devoted to the screening and evaluation of willow species for environmental phytoremediation.

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Optimization of Spirulina media by response surface methodology

XIE Lidan, WANG Suying

2017, 29(2): 307-314. DOI: 10.3969/j.issn.1004-1524.2017.02.18

Abstract ( 671 )

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To optimize the basic formula of Spirulina medium,the effects of soda ash production wastes, sodium nitrate(NaNO₃), potassium chloride (KCl), potassium dihydrogen phosphate (KH₂PO₄) on the phycocyanin content of Spirulina were determined using the response surface methodology (RSM) . Results showed that the optimal formula of Spirulina medium was soda ash production wastes 17 g, NaNO₃ 2 g, KCl 1 g, KH₂PO₄ 1.6 g, which can achieve 0.269 3 mg·L^-1 phycocyanin, 23% higher than that of the semi-synthetic medium used in outdoor cultivation.

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Evaluation of quality and safety of lightly salted Mylopharyngodon piceus in processing and storage

GUO Quanyou, DONG Yiwei, LI Baoguo, JIANG Zhaojun

2017, 29(2): 315-322. DOI: 10.3969/j.issn.1004-1524.2017.02.19

Abstract ( 466 )

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In order to investigate the quality and safety of lightly salted Mylopharyngodon piceus, changes in moisture content, water activity(Aw), pH, total viable counts (TVC), total volatile basic nitrogen (TVB-N) and thiobarbituric acid (TBA) during processing stages were studied, in addition to determination of muscle composition, mineral content and histamine etc. The results showed that pH increased initially in curing phase and then decreased from washing stage to drying stage, while moisture content, Aw, TVC, TVB-N and TBA kept decreasing during the whole processing stages. The moisture content, crude protein, crude fat and ash were (71.23 ± 1.16)%, (18.01 ± 0.34)%, (8.07 ± 0.08)% and (2.69 ± 0.06)%, respectively. The contents of essential amino acids (EAA) and delicious amino acids (DAA) were (5.74 ± 0.34)% and (6.90 ± 0.23)%, accounted for 36.17% and 40.83% of the total amino acids, respectively, meeting requirement of the balance style of essential amino acids ruled by FAO/WHO. However, unsaturated fatty acids/saturated fatty acid 3.43 and the total content of unsaturated fatty acids (77.4 ± 2.23)% suggested a high content of fat. Contents of microorganisms, histamine, heavy mental, TVB-N and TBA of lightly salted Mylopharyngodon piceus were acceptable in comparison to national standards.

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Preliminary studies on canning processing adaptability of citrus varieties cultivated in Quzhou area

JIA Yapeng, ZHANG Jun, LU Shengmin

2017, 29(2): 323-331. DOI: 10.3969/j.issn.1004-1524.2017.02.20

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Nowadays, Satsuma oranges were the main raw materials for the canned mandarins. In this work, nine citrus varieties mainly cultivated in Quzhou area of Zhejiang Province were selected to investigate their canning processing adaptability by comparative analysis on their anatomical characters (the difficulty of peeling, the difficulty of splitting, the edible ratio, the average seed number, the distortion ratio of slice and hardness of membrane), physicochemical properties (soluble solid, total sugar, total acid, vitamin C, total flavonoid, carotenoid and so on), and sensory evaluation after canning (juice transparency, slice uniformity, slice texture, and bitterness). The results indicated that the anatomical traits of Mantouhong, Nanxiang, Shanxiahong, seedless Shanxiahong Lipeng No.2 and Weizhang were suitable for canning, the canned seedless Shanxiahong, Weizhang, Nanxiang, Lipeng No.2, Shanxiahong had the higher sensory score, indicating that in addition to the Satsuma orange strain, Ponkan (Lipeng No.2) and citrus hybrid (Nanxiang) are also suitable for processing into canned mandarin.

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Heterologous expression and purification of plantaricin JK and analysis of its antibacterial activity

JIANG Han, LIU Jiao, LI Ping, GU Qing

2017, 29(2): 332-337. DOI: 10.3969/j.issn.1004-1524.2017.02.21

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In order to obtain the high-yield two-peptide bacteriocin plantaricin JK, pln J and pln K were successfully heterologously expressed in Escherichia coli BL21 (DE3), and were induced by IPTG. Then the two peptides were expressed as His₆-tag fusion proteins and were separated by Ni²⁺ chelating affinity chromatography. The fusion proteins were cleaved by enterokinase and further purified. The yields of pln J and pln K were around 2-3 and 5-8 mg·L^-1. The antibacterial activity of the peptides against 4 indicator strains, i.e. Staphylococcus citreus, Escherichia coli, Micrococcus luteus, Listeria monocytogenes, was analyzed by agar diffusion method. It was shown that pln J and pln K both could inhibit the growth of the 4 indicator strains, but plantaricin JK had larger inhibition zone than pln J or pln K alone.

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Plant classification method based on dictionary learning with sparse representation

ZHANG Shanwen, KONG Weiwei, WANG Zhen

2017, 29(2): 338-344. DOI: 10.3969/j.issn.1004-1524.2017.02.22

Abstract ( 475 )

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Plant classification based on leaf image is an important research area in plant taxonomy. Because the leaf image is complex and is sensitive to the season and illumination, the classification results of the existing plant classification methods are not robust. Based on the dictionary learning with sparse representation, a plant classification method was proposed in this paper. The plant classification problem was transformed to solve the sparse representation problem of the test sample to the training samples. A small optimal over-complete dictionary was designed to calculate the sparse representation of the leaf image by using the class-specific dictionary learning. Comparing to the other methods, the proposed method didn't need to extract the features of color, texture and shape of the leaf image. So the computing complexity was reduced and the robustness and the real-time performance of the automatic identification of plant were improved. The experimental results on the real-world database of 50 kinds of the plant leaf images showed the feasibleness of the proposed algorithm. The recognition rate was more than 92%.

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Cloning and encoding protein bioinformatics analysis of BdCDPK4 from Brachypodium distachyon

WEI Shuya, ZHAO Xudong, WANG Huiping, YANG Guangxiao, HE Guangyuan

2017, 29(2): 345-352. DOI: 10.3969/j.issn.1004-1524.2017.02.23

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Brachypodium distachyon, the genome of which shares high homology with wheat and barley, has been used as the new model species of Gramineae. In order to clarify the structure and function of Brachypodium distachyon CDPKs gene, the coding region sequence (CDS) of BdCDPK4 gene of Brachypodium distachyon was isolated, and its character, structure and function were analyzed by bioinformatics methods. The results showed that 1 851 bp cDNA (GenBank ID: XM_003559516) of coding sequence of BdCDPK4 gene included the complete 1 752 bp CDS, coded 583 amino acids, and located on chloroplast and vacuole. The molecular weight and isoelectric point of BdCDPK4 protein were 64.78 ku and 9.12, respectively. BdCDPK4 protein contained several domains including N-variable domain, kinase domain, autoinhibitory domain, EF-hands and CaM-like domain. Phylogenetic analysis results suggested that BdCDPK4 protein had high homology with wheat TaCDPK19. TaCDPK19 were induced by high salt and white powder Blumeria graministritici(Bgt). BdCDPK4 protein may participate in the regulation of signal transduction pathways under stresses according to the homologies of protein domain among all species. Taken together, our data could establish a good foundation for studying the biological functions of BdCDPK4 gene.

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