龙眼DCL4的亚细胞定位分析及其对外源ABA和SA的响应

为了研究DlDCL家族中DlDCL4基因在龙眼中的功能,本研究通过氨基酸序列比对、系统发育进化树的构建、亚细胞定位验证、实时荧光定量PCR（qPCR）等方法,对其进行初步探究。结果表明：遗传进化分析显示,DCL4氨基酸序列在所选的物种间相似度高于50%,其中龙眼与甜橙（Citrus sinensis）的亲缘关系最近。在激光共聚焦显微镜观察下,洋葱细胞核发出绿色荧光,其他部位不发出荧光,表明龙眼DCL4蛋白定位于细胞核中。在24 h内,100 μmol/L的外源脱落酸（ABA）对 DlDCL4都有极显著的下调效果;100 μmol/L的水杨酸（SA）对其表达量有显著的上调效果,其中8 h时表达量最大,提示 DlDCL4对不同激素的响应效果不同且不同处理时长下响应效果也会有影响。     

菜籽油裂解燃料离子液体催化酯化反应降酸工艺的研究

为了降低裂解燃料酸值,增加燃料的稳定性能,将吡啶丁烷磺酸硫酸氢盐应用于菜籽油裂解燃料的酯化反应。考察了催化剂用量、反应时间和反应温度等对酸值的影响,并在最佳优化条件下考察了裂解燃料成品的理化性质。结果表明:吡啶丁烷磺酸硫酸氢盐对催化酯化反应具有很高的催化活性。优化工艺条件为:催化剂用量1.2%、反应温度75℃、反应时间70 min,在此工艺条件下,酸值降低到1.0 mgKOH/g以下。     

稳定碳、氮同位素在牛肉加工过程中的分馏效应

为了探究牛肉及副产物中稳定碳、氮同位素在加工过程中的变化规律,确证稳定碳、氮同位素在牛肉加工制品产地溯源中的稳定性和有效性。本试验通过对牛肉进行不同时间的水煮、烤制和油炸3种处理,其中水煮和烤制加工时间分别为5、10、15、20、25和30 min,油炸加工时间分别为1、2、3、4和5 min;采用元素分析仪-稳定同位素比率质谱仪(EA-IRMS)测定脱脂牛肉、粗脂肪及副产物中δ¹³C和δ¹⁵N值。结果表明,脱脂牛肉中δ¹³C值在水煮、烤制和油炸3种加工方式不同加工时间之间均无显著差异,水煮和烤制后粗脂肪中δ¹³C值无显著变化,油炸后的牛肉粗脂肪δ¹³C值主要受植物油的影响,加工方式及加工时间对其无显著影响;水煮脱脂牛肉δ¹⁵N值在加工25、30 min时与对照组牛肉存在显著差异,但平均差值仅为0.3‰~0.9‰。牛肉稳定碳、氮同位素在不同加工过程中分馏效应较小,可有效用于牛肉加工半成品及成品的原产地溯源。     

Intensive organic vegetable production increases soil organic carbon but with a lower carbon conversion efficiency than integrated management

Intensive vegetable production in greenhouses has rapidly expanded in China since the 1990s and increased to 1.3 million ha of farmland by 2016, which is the highest in the world. We conducted an 11‐year greenhouse vegetable production experiment from 2002 to 2013 to observe soil organic carbon (SOC) dynamics under three management systems, i.e., conventional (CON), integrated (ING), and intensive organic (ORG) farming. Soil samples (0–20 and 20–40 cm depth) were collected in 2002 and 2013 and separated into four particle‐size fractions, i.e., coarse sand (> 250 µm), fine sand (250–53 µm), silt (53–2 µm), and clay (< 2 µm). The SOC contents and δ¹³C values of the whole soil and the four particle‐size fractions were analyzed. After 11 years of vegetable farming, ORG and ING significantly increased SOC stocks (0–20 cm) by 4008 ± 36.6 and 2880 ± 365 kg C ha^?1 y^?1, respectively, 8.1‐ and 5.8‐times that of CON (494 ± 42.6 kg C ha^?1 y^?1). The SOC stock increase in ORG at 20–40 cm depth was 245 ± 66.4 kg C ha^?1 y^?1, significantly higher than in ING (66 ± 13.4 kg C ha^?1 y^?1) and CON (109 ± 44.8 kg C ha^?1 y^?1). Analyses of ¹³C revealed a significant increase in newly produced SOC in both soil layers in ORG. However, the carbon conversion efficiency (CE: increased organic carbon in soil divided by organic carbon input) was lower in ORG (14.4%–21.7%) than in ING (18.2%–27.4%). Among the four particle‐sizes in the 0–20 cm layer, the silt fraction exhibited the largest proportion of increase in SOC content (57.8% and 55.4% of the SOC increase in ORG and ING, respectively). A similar trend was detected in the 20–40 cm soil layer. Over all, intensive organic (ORG) vegetable production increases soil organic carbon but with a lower carbon conversion efficiency than integrated (ING) management.     

Early resynchronization protocols for goats: Progestogens can be used prior to an early pregnancy diagnosis without affecting corpus luteum functionality

This study aimed to evaluate the exogenous progesterone (P4) effect on the luteal function from Day 16 to Day 21 of the oestrous cycle in inseminated goats with unknown pregnancy status. A total of 54 does passed through a short progestin-based synchronization protocol and, on Day 16 of the following oestrous cycle, 27 does received a new P4 device which was retained until Day 21. Blood samples were collected daily from all does during this period, as well as on Day 24. Pregnancy diagnoses were performed on Day 30. Serum P4 values from 26 animals (G_NPSP: Group of non-pregnant does with second sponge: n = 8; G_NPNSP: Group of non-pregnant does without second sponge: n = 6; G_PSP: Group of pregnant does with second sponge: n = 5; G_PNSP: Group of pregnant does without second sponge: n = 7) were determined by radioimmunoassay commercial kits. No P4 differences were found between groups (G_NPSP: 3.1 ± 2.8; 1.7 ± 1.8; 0.4 ± 1.0; and 0.0 ± 0.0 vs. G_NPNSP: 4.4 ± 1.8; 3.0 ± 2.2; 0.8 ± 0.8; and 0.0 ± 0.0 or G_PSP: 4.2 ± 1.0; 3.4 ± 0.6; 3.3 ± 1.6; 3.2 ± 0.9; 3.6 ± 1.2; 3.5 ± 1.3; 2.7 ± 1.3 vs. G_PNSP: 4.4 ± 1.6; 3.6 ± 1.5; 3.7 ± 1.5; 3.8 ± 1.4; 3.2 ± 1.2; 3.1 ± 1.2; 3.6 ± 1.1; D16, D17, D18, D19, D20, D21, D24, respectively) or for the interaction of group and time. In conclusion, a second progestogen device had no effect on luteolysis or early pregnancy in the following oestrous cycle.     

122份栽培桃品种（系）黄白肉性状的分子标记辅助鉴定

【目的】 类胡萝卜素裂解双脱氧酶基因（Carotenoid Cleavage Dioxygenases 4,CCD4）控制桃果肉颜色（白/黄）,CCD4存在3种等位基因。本研究利用Indel、SSR荧光标记毛细管电泳及SNP鉴定等基因分型技术分析我国主要桃黄白肉品种（系）中CCD4等位基因的差异,为主要黄/白肉品种（系）的基因型鉴定、亲本选配和选择相应的分子标记对不同来源子代的果肉颜色进行鉴定奠定基础。【方法】利用已经报道的桃不同果肉颜色中CCD4等位基因3种突变类型,合成不同引物进行PCR扩增,LTR反转录转座子插入突变经1%的琼脂糖凝胶电泳检测,CT单元重复的PCR产物在ABI3730XL测序仪上进行SSR荧光标记毛细管电泳检测,SNP标记经Sanger测序后利用ContigExpress软件分析CCD4等位基因的碱基替换（A→T）。综合以上结果,统计每份材料中CCD4等位基因的突变类型与果肉颜色的一致性。【结果】通过对不同来源的122份桃品种（系）材料进行基因型分析,发现CCD4发生LTR反转录转座子插入突变材料的基因型共有31份,占总材料的25.4%,其中纯合插入突变材料的片段扩增长度为729 bp,共有8份,占总突变的25.8%;CCD4发生微卫星重复序列突变材料存在2 bp的插入,扩增片段长度为179 bp,该类型共有68份,占总材料的55.7%,其中纯合插入材料25份,占总突变的36.8%;CCD4发生A→T碱基替换突变的材料较少,仅有1份,占总材料的0.82%,实际应用中可以不考虑该种类型。CT和LTR插入的两种突变类型的黄肉品种（系）有7份,占总材料的5.7%。研究结果表明,LTR反转录转座子插入突变和微卫星序列重复突变是黄肉桃中CCD4等位基因的主要突变类型。其中CCD4发生一种纯合突变或两种杂合突变桃品种（系）为黄肉类型,分子标记鉴定结果与调查的122份桃品种（系）黄白肉表型性状完全一致,准确率为100%。【结论】采用分子标记明确了122个桃品种（系）黄/白肉性状的基因型,为不同亲本组合子代表型鉴定的标记类型选择提供了技术支撑,为建立桃种质材料黄/白性状的分子辅助育种体系和黄肉桃的选育奠定了基础。     

秸秆还田与优化施氮改善烤烟根际土壤碳氮转化及氮素吸收

针对华南沙泥田烟区氮肥过量施用导致烟叶品质下降的问题,拟通过秸秆还田与氮肥优化配施调节烤烟各生育期氮吸收、根际土壤碳氮转化及相关酶的活性。试验采用随机区组设计,设水稻秸秆还田与氮肥两因素,其中3个碳（秸秆还田）水平：C0,无秸秆还田,0 kg/hm ²;C1,中量秸秆还田,4500 kg/hm ²;C2,全部秸秆还田,9000 kg/hm ²;2个氮水平：N1,传统施氮,169.50 kg/hm ²;N2,优化施氮,105.00 kg/hm ²。结果表明,与N1相比,N2处理秸秆还田下烤烟前期烟叶和根系氮吸收和累积正常,而后期有所下降;C1、C2处理时,与现蕾期相比,成熟期时根系氮含量均降低4.2 g/kg,叶片氮含量降低3.8、3.0 g/kg;成熟期时,与N1处理相比,N2处理时C1、C2处理的土壤无机氮含量降低30.4%、20.0%,前期2个氮处理间土壤无机氮含量无差异,优化施氮下中量秸秆还田（C1N2）处理烤烟土壤NH₄ ⁺-N含量在整个生育期内较高且呈降低趋势,而硝态氮和土壤无机氮含量在成熟期最低,表明中量秸秆还田下优化施氮能够维持生育前期土壤无机氮含量,且降低成熟期土壤无机氮含量。2个秸秆还田量处理时,团棵期N2处理的土壤转化酶活性是N1处理的1.30、1.13倍;旺长期土壤转化酶活性是N1处理的1.27、1.10倍;与传统施氮相比,秸秆还田下优化施氮处理增加了整个生育期土壤脲酶、磷酸酶、过氧化氢酶活性。在中量秸秆还田（C1）处理下,与N1水平相比,现蕾期和成熟期的N2处理土壤水溶性碳含量提高11.2%和14.1%;与N1相比,N2处理的土壤微生物量氮含量呈降低趋势。综上所述,优化施氮耦合中量秸秆还田（C1N2）能够提高土壤水溶性碳含量,维持土壤氮素合理供应,稳定烤烟生育前期对氮素吸收与累积,减少生育后期烟叶氮素奢侈吸收,与烤烟吸氮规律比较吻合。     

miR-106b-5p靶向KLF4调控山羊肌内前体脂肪细胞分化

旨在明确miR-106b-5p对山羊肌内前体脂肪细胞分化的影响，并确定这种作用是通过靶向KLF4来实现的。本研究利用实时荧光定量PCR （quantitative real-time PCR，qRT-PCR）技术检测miR-106b-5p在山羊肌内前体脂肪细胞分化过程中的表达模式，通过脂质体转染技术将miR-106b-5p mimic和miR-106b-5p inhibitor转入体外培养的山羊肌内前体脂肪细胞，油红O染色法从形态学验证miR-106b-5p对脂肪细胞中脂滴积聚的影响，qRT-PCR检测预测的靶标基因KLF4和脂肪分化标志基因的表达情况，利用双荧光素酶报告系统鉴定miR-106b-5p与KLF4的靶标关系。qRT-PCR结果显示，miR-106b-5p在山羊肌内前体脂肪细胞诱导分化第3天时表达量最高。在山羊肌内脂肪细胞中干扰miR-106b-5p后油红O染色显示脂滴聚积减少，过表达miR-106b-5p后脂滴聚积增加。在山羊肌内前体脂肪细胞中转染miR-106b-5p inhibitor后PPARγ表达量显著降低（P<0.05），而KLF4表达量极显著升高（P<0.01）；转染miR-106b-5p mimic后LPL和PPARγ表达量极显著升高（P<0.01）。荧光素酶活性试验结果显示，过表达miR-106b-5p可显著抑制KLF4荧光活性。miR-106b-5p通过靶向并负调节KLF4的表达促进山羊肌内脂肪细胞分化。     

Cytokine profiles in peripheral blood mononuclear cells from patients with cystic echinococcosis

IntroductionCystic echinococcosis (CE) is a chronic zoonotic disease caused by the larval stage of Echinococcus granulosus (E. granulosus), which affects domestic and wild carnivores as the definitive host and ungulates as intermediate hosts. In intermediate hosts, both Th1 and Th2 cells are involved in the immune responses to an echinoccocal infection. This study aimed to investigate production of IL-4, IL-10, and IFN-γ cytokines in peripheral blood mononuclear cells (PBMCs) of CE patients before and after surgical treatment.MethodsTo evaluate cytokine production in response to E. granulosus antigens, we investigated IL-4, IL-10, and IFN-γ production in PBMCs of 20 CE patients in response to hydatid cyst fluid antigen (HCF-Ag) before and after surgical treatment using ELISA.ResultsThe mean IL-4 production from HCF-Ag stimulated PBMCs was significantly decreased (p < 0.05), while IFN-γ was significantly increased in HCF-Ag stimulated PBMCs in patients after surgery (p = 0.005).Furthermore, our results showed that there is no significant difference between IL-10 production in patients before and after treatment (p = 0.562).ConclusionsOur data Indicated production of IL-4 in cultured PBMCs of CE patients stimulated with HCF-Ag was decreased significantly. While, production of IFN-γ was increased significantly in responses to HCF Ag after surgery. We concluded that the evaluation of IL-4 and IFN-γ in HCF-Ag stimulated PBMCs of CE patients should be considered as a useful marker in the follow up of patients with cystic echinococcosis.