'寒富'苹果干简易加工技术探讨与营销分析

以'寒富'苹果为试材,采用热风干燥制作方法,分别用60、70和80℃烘干处理6、8、10h,烘干后进行真空包装、带封口塑料袋包装、保鲜袋包装和不包装处理,探究苹果干简易加工方法及分析销售前景.结果 表明,冬季'寒富'苹果去皮后在70℃条件下烘干10 h制成的苹果干软硬、酸甜、色泽和口感均好于其他处理,夏季80C条件下烘干10h效果略好于其他处理,但整体口感不如冬季处理.在包装上,简易真空包装对于保持果干质量、延长销售时间等效果好于其他处理.城市近郊观光采摘果园、专业合作社采用优化后的方法加工制作苹果干,作为待客零食及乡土地域特色产品销售,具有较大的增加果园收益空间.     

木枣抗裂变异系的鉴定和果实性状分析

在枣品种的抗裂性调查中,发现“木枣”存在变异系。本研究以“木枣”、变异系“V1”、变异系“V2”和易裂品种“狗头枣”为供试材料,在采用冷水浸泡法进行室内裂果率测定和抗裂性分析的基础上,对单果重、果纵茎、果横茎、果纵横茎比、单核重、核纵茎、核横茎、核纵横茎比、梗洼宽度、肉核比、含水量等果实性状进行了测定。结果表明：裂果率表现为狗头枣>V1≈木枣>V2,筛选出木枣优良抗裂变异系V2,与木枣遗传背景一致,抗裂性又极显著优于木枣,可为后续裂果机理研究和抗裂品种选育提供育种材料。品种的抗裂性与果形、核形、肉核比、梗洼宽度有关。果实较小、果核较小、果形细长、肩大尾尖、肉核比高、梗洼宽度大,抗裂性好。     

Cloning,Expression and Bioinformatics Analysis of Enterococcus faecalis from Northeast Black Bear

In order to detect Enterococcus faecalis endocarditis antigen (EfaA) of bear that was used to disclose the infected Northeast Black bear immediately, a pair of PCR primers was synthesized by EfaA gene of Enterococcus faecalis that was recorded in GenBank (accession number:U03756.1) and the target fragment was 689 bp. The PCR product was cloned by pMD19-T vector, then was transferred to Escherichia coli DH5α competent cells and the positive clones were filtered. Recombinant plasmid (pMD19-T-EfaA) was extracted. Identification of PCR and digestion, sequencing, the structure prediction of proteins were operated. The results showed that EfaA gene was cloned successfully, and the gene homology with ATCC 29212 was 100.0%. pMD19-T-EfaA was constructed successfully, structure of proteins was predicted. This study provided theoretical basis for diagnostic methods of Enterococcus faecalis and laid basis for prevention and control of Northeast Black bear disease.     

蚯蚓粪益生菌肥对铁棍山药不同生长时期 共生微生物群落及其品质的影响

蚯蚓粪富含有机质、腐殖酸及微生物菌群,可作为植物养分供应和土壤理化性质改善的常用有机肥基质,然而,其对山药的生长和品质的影响尚不清楚。以木质纤维素饲喂的赤子爱胜蚓得到的蚯蚓粪为基质配施益生菌剂开展山药施用效果的研究,统计分析蚯蚓粪益生菌肥添加对山药生长和品质的变化,探究其对于山药共生微生物群落的影响。结果表明:蚯蚓粪益生菌肥的使用促进了铁棍山药的株高及单株重,提高了山药的产量,收获期铁棍山药中黄酮含量升高了2.48 mg/g,增长约37.48%,可溶性糖、蛋白质及皂苷等营养成分分别提高0.26、0.14、0.31 倍;通过对铁棍山药不同生长时期根际微生物和内生微生物群落结构的分析表明,施肥后根际土壤中链霉菌、鞘氨醇单胞菌、芽孢杆菌明显增多,绿僵菌等明显减少,并且这些丰度明显改变的微生物也与土壤理化性质之间有着明显的相关性。内生微生物中泛菌、枝孢菌明显增多,青枯菌、Cyphellophora 等病原微生物丰度明显减少。蚯蚓粪益生菌肥的施用改善了山药根际微生态健康状况,减少了发病率,提高了山药品质。     

栀子叶斑病及其病原的鉴定

首次报道在栀子(Gardenia jasminoides)上存在由拟盘多毛孢属真菌引致的叶斑病。从湖北省长阳县采集到栀子叶斑病病害标本,并利用柯赫氏法则证实该病由真菌GJ-1菌株所引致;根据GJ-1菌株的形态学特征和ITS序列的分析、比较,证实GJ-1菌株属于拟盘多毛孢属(PestalotiopsisSteyaert)真菌;GJ-1菌株的ITS序列与P.gracilis的ITS同源性达100%,因此推定它属于P.gracilis。     

OCX-32基因内含子变异对长顺绿壳蛋鸡蛋壳品质的影响

[目的]探讨OCX-32基因对长顺绿壳蛋鸡蛋壳品质的遗传效应,为长顺绿壳蛋鸡的保种选育及开发利用提供参考依据.[方法]以长顺绿壳蛋鸡为材料,采用PCR产物直接测序法筛选OCX-32基因SNP多态位点,实时荧光定量PCR检测组织表达谱,运用SPSS 19.0广义线性模型(GLM)分析SNP位点基因型或双倍型与所测定性状指标的相关性.[结果]在长顺绿壳蛋鸡OCX-32基因中检测到3个SNPs位点,分别是位于内含子1上的g.22592323G>T及内含子3上的g.22597350T>C和g.22597555G>A.卡方(χ2)检测结果发现,g.22597350T>C位点的基因型分布显著偏离Hardy-Weinberg平衡(P<0.05,下同).连锁不平衡分析结果显示,3个SNPs突变位点不存在强连锁不平衡,共发现4种单倍型和8种双倍型,单倍型H1和双倍型H1H2频率最高,分别为0.523和0.345.实时荧光定量PCR检测结果显示,OCX-32基因在长顺绿壳蛋鸡12个组织中存在不同程度的表达,表达量排序为肾脏>心脏>子宫>腹脂>小肠>脾脏>肝脏>腺胃>胸肌>胰腺>肺脏>肌胃.关联分析结果显示,g.22597350T>C位点CC基因型在蛋壳强度和蛋壳重2个品质指标上显著高于CT基因型和TT基因型;双倍型H4H4个体的蛋壳强度显著高于其他7种双倍型个体,蛋壳重显著高于H2H4个体.[结论]OCX-32基因内含子变异能影响长顺绿壳蛋鸡蛋壳品质,检测到的3个SNPs位点可作为鸡蛋壳品质选择的遗传分子标记,其中突变位点g.22597350T>C的CC基因型和双倍型H4H4是影响蛋壳强度和蛋壳重的关键基因型和双倍型,有利改善蛋壳品质.     

A novel glycoside hydrolase 74 xyloglucanase CvGH74A is a virulence factor in Coniella vitis

Grape white rot is a destructive fungal disease occurring worldwide. Recently, Coniella vitis was identified as the predominant pathogen causing this disease in China. As the periderms of grape shoots are severely degraded by C. vitis, it was speculated that cell wall-degrading enzymes (CWDEs) might play a key role in the pathogenesis of this disease. Therefore, this study aimed to examine the hydrolytic activity of the CWDEs of C. vitis. The results showed that xylanase (Xy) and xyloglucanase (XEG) had high levels of hydrolytic activity both in vitro and in vivo. Furthermore, a high-virulence fungal strain exhibited higher levels of Xy and XEG activities compared with a low-virulence strain. The genome of the fungus was found to harbor two XEG-coding genes CvGH74A and CvGH74B, which belonged to the glycoside hydrolase (GH)74 family. The expression level of CvGH74A was found to be high during pathogen infection. CvGH74A gene deletion mutants were generated using the split-marker method. The deletion of CvGH74A decreased both the hydrolytic activities of XEG and Xy and also the ability of the fungus to infect the grape leaves. No differences in the hyphal growth, morphology of colonies, or conidiation were found between the ΔCvGH74A mutant strains and the wild-type strain. Together, these results suggested that CvGH74A acted as an important virulence factor, and its enzymatic activity might regulate the virulence of the pathogen. This study was novel in reporting that GH74 XEG acted as a virulence factor in C. vitis.     

Long-term fertilization leads to specific PLFA finger-prints in Chinese Hapludults soil

Soil microbes play essential roles in the biogeochemical processes of organic carbon and nutrient cycling. Many studies have reported various short-term effects of fertilization on soil microbes. However, less is known about the effects of longterm fertilization regimes on the rhizosphere. Therefore, the objective of this study was to explore how the soil microbial communities in the rhizosphere respond to different long-term fertilization strategies. Based on a 21-year field treatment experiment in Guizhou, China, we extracted phospholipid fatty acids(PLFAs) to determine the microbial community structure in both the non-rhizosphere(NR) and rhizosphere(R). Six treatments were included: no fertilizer(CK), mineral nitrogen fertilizer(N), N with potassium(NK), phosphorus with K(PK), NPK, and NPK combined with manure(MNPK). The results showed that total PLFAs under unbalanced mineral fertilization(N, NK and PK) were decreased by 45% on average in the NR compared with CK, whereas MNPK increased fungi and G~– bacteria abundance significantly in both the NR(by 33 and 23%) and R(by 15 and 20%), respectively. In addition, all microbial groups in the R under these treatments(N, NK and PK) were significantly increased relative to those in the NR, except for the ratio of F/B and G~+/G~–, which might be due to the high nutrient availability in the R. Soil pH and SOC significantly regulated the soil microbial community and structure, explaining 51 and 20% of the variation in the NR, respectively. However, the rhizosphere microbial community structure was only significantly affected by soil pH(31%). We concluded that the soil microbial community in the NR was more strongly affected by long-term fertilization than that in the R due to the rhizosphere effect in the agricultural ecosystem. Rhizosphere nutrient conditions and buffering capacity could help microbial communities resist the change from the long-term fertilization.     

Minimally modified low-density lipoprotein up-regulates endothelin receptors in mouse mesenteric artery through p38 MAPK pathway

AIMTo investigate whether minimally modified low-density lipoprotein (mmLDL) affects the quantity and activity of endothelin （ET） type A (ET_A) and type B (ET_B) receptors in mouse mesenteric artery by activating p38 mitogen-activated protein kinase (MAPK) inflammatory pathway. METHODSThe KM mice were divided into normal saline (NS) group (injection of NS via caudal vein), mmLDL group (injection of mmLDL via caudal vein), LDL group (injection of LDL via caudal vein), mmLDL+SB 203580 group (injection of mmLDL via caudal vein and intraperitoneal injection of p38 MAPK pathway specific inhibitor SB 203580) and mmLDL+DMSO group (injection of mmLDL via caudal vein and intraperitoneal injection of DMSO). Mesenteric artery ring segment vasoconstriction dose-response curves affected by sarafotoxin 6c (S6c) and ET-1 were recorded by the myography system. The mRNA levels of ET_B receptor, ET_A receptor and interleukin-6 (IL-6) were detected by RT-qPCR. The protein levels of ET_B receptor, ET_A receptor, IL-6, p38 MAPK, p-p38 MAPK, NF-κB and p-NF-κB were determined by Western blot. The serum concentration of IL-6 was measured by ELISA. RESULTSThe contractile responses of the blood vessel segments to S6c and ET-1 were significantly increased by mmLDL (P<0.01). The mRNA and protein expression levels of ET_A receptor, ET_B receptor, and IL-6 significantly increased (P<0.01). The protein levels of p-p38 MAPK and p-NF-κB were significantly increased (P<0.01). The serum level of IL-6 was significantly increased (P<0.01). These effects of mmLDL were inhibited by p38 MAPK inhibitor SB 203580. CONCLUSION mmLDL increses the serum concentration of IL-6, up-regulates the expression of IL-6, ET_A receptor and ET_B receptor in mouse mesenteric artery, and enhances the vasoconstriction function medi?ated by ET_A and ET_B receptors, which is related to the activation of p38 MAPK inflammatory pathway and downstream NF-κB pathway.     

Roles of protein phosphatase 4 in down-regulation of eNOS Ser633 phosphorylation induced by palmitic acid in human umbilical vein endotheli?al cells

AIMTo investigate the roles of protein phosphatase 4 (PP4) in down-regulation of endothelial nitric oxide synthase (eNOS) Ser633 phosphorylation induced by palmitic acid (PA). METHODSHuman umbilical vein endothelial cells (HUVECs) were treated with PA at 25 μmol/L, 50 μmol/L, 100 μmol/L and 200μmol/L for 36 h, or treated with PA at 100 μmol/L for 12 h, 24 h, 36 h and 48 h. Protein phosphatase 2A (PP2A) family inhibitor fostriecin (FST, 20 nmol/L) or okadaic acid (OA, 5 nmol/L) was selected to pretreat the HUVECs for 30 min. Protein phosphatase 4 catalytic subunit (PP4c) siRNA or protein phosphatase 2A catalytic subunit (PP2Ac) siRNA was transfected into the HUVECs. The protein expression levels of of eNOS, PP4c and PP2Ac, as well as the level of eNOS Ser633 phosphorylation, were detected by Western blot. The intracellular nitric oxide (NO) content was measured by DAF-FM DA. RESULTS(1) Compared with control group, the levels of eNOS Ser633 phosphorylation were decreased in PA groups in which the HUVECs were treated with 25 μmol/L, 50 μmol/L, 100 μmol/L and 200 μmol/L PA for 36 h (P<0.05) and 100 μmol/L PA for 24 h, 36 h and 48 h (P<0.05). No significant difference in the level of total eNOS protein expression among all the groups was observed. (2) Compared with control group, both FST and OA pretreatment reversed the reduction of eNOS Ser633 phosphorylation (P<0.05) and the decrease in intracellular NO content (P<0.05) induced by PA. No significant difference in the level of total eNOS protein expression among all the groups was observed. (3) Compared with si-Control group, the PP4c protein expression was significantly reduced (P<0.05), while the level of eNOS Ser633 phosphorylation was significantly increased in si-PP4c group (P<0.05). Although the levels of PP2Ac protein expression declined significantly (P<0.05), the level of eNOS Ser633 phosphorylation remained unchanged in si-PP2Ac group. No significant differencein the level of total eNOS protein expression among all the groups was found. CONCLUSION PA significantly reduces the level of eNOS Ser633 phosphorylation and the content of NO in the HUVECs, which may be due to PA inducing the activation of the PP2A family member PP4 rather than PP2A.