MSTN基因编辑牛肌肉转录组测序及生物信息学分析

为探究肌生长抑制素（myostatin,MSTN）基因对牛肌肉发育的具体调控机制,本研究选取同一牛场健康鲁西黄牛10头,其中通过转基因技术得到的基因编辑牛MSTN^-/-和同种非转基因野生型牛各5头。分别采集两组牛腿臀肌肉样品,利用IIlumina HiSeq高通量测序技术进行转录组测序分析,通过生物信息学方法比较两组样本间的差异表达基因,并进行GO和KEGG富集分析,最后利用实时荧光定量PCR验证转录组测序数据。结果显示,基因编辑型牛和野生型牛之间共检测到18 071个基因。在log₂|FoldChange|≥ 1.48条件下,筛选出406个差异表达基因,其中347个显著上调,59个显著下调。GO功能富集分析显示,MSTN基因编辑后显著影响915个功能类别（P<0.05）,差异基因主要参与结合、生物系统调节、免疫系统等相关功能。KEGG通路富集分析结果共涉及211个通路,差异基因主要富集在细胞黏附分子、趋化因子信号通路、细胞因子互作等信号通路上,进一步从中筛选出可能参与细胞生长、肌肉发育的差异基因（CD14、KIT、CSF1R、FBP1、DUSP4、ULBP21、PRKCB、SPN、CHAD、SRC）。实时荧光定量PCR检测结果显示,所选差异基因表达水平与转录组表达水平一致,证明测序结果的可靠性。本研究结果表明,MSTN基因发挥作用后可以介导多个下游基因表达,从而影响相关信号通路及生物学过程;同时,所筛选出的差异表达基因可作为进一步研究骨骼肌调控机制的候选靶标。     

MSTN基因对牛肌动蛋白细胞骨架调节通路的影响

为探究肌肉生长抑制素（myostatin,MSTN）对牛骨骼肌生长发育的作用机制,本研究前期利用定量蛋白质组学与磷酸化蛋白质组学分析野生型蒙古牛（MG.WT）和MSTN^+/-蒙古牛（MG.MSTN^+/-）腿臀肌肌肉组织中蛋白质水平和磷酸化修饰水平的差异变化,使用已建立的牛骨骼肌卫星细胞体外诱导成肌分化模型,检测设计合成的MSTN siRNA （si-MSTN）干扰效果;采用实时荧光定量PCR和Western blotting方法检测转染si-MSTN的增殖期（GM）和分化第3天（DM3）牛骨骼肌卫星细胞中肌动蛋白细胞骨架调节通路相关基因的mRNA和蛋白水平的表达变化,研究敲低MSTN表达对肌动蛋白细胞骨架调节通路的影响。结果显示,在MSTN^+/-蒙古牛肌肉组织中共鉴定到16个肌动蛋白细胞骨架调节通路相关基因表达丰度上调;转染si-MSTN细胞中的MSTN表达水平极显著降低（P<0.01）;在转染si-MSTN的GM期牛骨骼肌卫星细胞中,肌动蛋白细胞骨架调节通路相关基因ENAH、ACTN4和Cdc42的mRNA水平均显著升高（P<0.05）,PFN1、RhoA和ACTN4的蛋白水平均显著或极显著升高（P<0.05;P<0.01）;在转染si-MSTN的DM3牛骨骼肌卫星细胞中,ENAH、CFL1、SCIN和Cdc42基因mRNA水平均显著升高（P<0.05）,RhoA基因mRNA水平极显著升高（P<0.01）,PFN1和ACTN4的蛋白水平均显著升高（P<0.05）。结果表明,干扰MSTN可以促进肌动蛋白细胞骨架调节通路相关基因的表达,探明了MSTN可能通过介导肌动蛋白细胞骨架调节通路影响牛骨骼肌卫星细胞增殖和成肌分化的分子机制,为进一步研究MSTN对牛成肌分化的调控机制提供参考。     

苯并芘对猪卵母细胞SHH信号通路影响的研究

如今空气污染问题仍广泛存在,其中苯并芘（Benzo （a） pyrene,BaP）是一种常见的空气污染物,对人和动物的健康具有严重影响。据报道,BaP可以显著降低卵母细胞IVM （卵母细胞体外成熟）能力,但机制尚不清楚。因此,本研究在猪卵母细胞IVM培养液中添加不同浓度BaP （50 μM、100 μM、200 μM）检测其对卵母细胞IVM能力的影响,并检测此过程SHH信号通路相关基因和蛋白的表达。结果显示50 μM BaP处理组卵母细胞IVM能力显著低于对照组（76.5 vs 68.1,P<0.05）,并随着BaP浓度增加进一步下降。同时,qPCR和免疫荧光染色检测发现,50 μM BaP处理组卵母细胞中SHH信号通路相关基因mRNA和蛋白表达量,均显著低于对照组（P<0.05）。因此,本研究初步证明BaP可能通过破坏SHH信号通路降低猪卵母细胞IVM能力。     

头骨形态指标鉴别水貂来源有效性的评估

人工驯养野生动物是获取野生动物产品的重要手段,但是非法盗猎的野生动物产品以人工饲养的名义进入市场,就会对野生动物的保护形成挑战。因此,准确鉴别市场上的动物产品是来自养殖场还是野外,是保护野外种群,维持养殖业正常秩序的关键。本研究以北美水貂(Neovison vison)为例,基于颧骨内侧空间(ZAS)、颧宽(ZAA和ZBA)、颅基长(BSL)、下颌长(MDL)、眶后收缩(POC)和最大面部宽度(GFB)等量度,建立了9个头骨形态计量学指标:POC/GFB、POC/BSL、POC/MDL、ZAS SQRT/BSL、ZA SSQRT/MDL、ZBA×BSL/2ZAS、ZBA×MDL/2ZAS、ZBB×BSL/2ZAS和ZBB×MDL/2ZAS。野生组(n=32,9雄23雌)和饲养组(n=45,35雄10雌)的比较表明,除POC/GFB外,所有指标对于在两组样品之间均有显著差异,整体判别正确率为73.2%—85.5%。在性别已知的情况下,雄性的整体正确率为82.4%—93.8%,雌性为41.7%—85.7%。为此,应针对不同性别、不同来源的动物建立相应的参考数据,利用似然比进行鉴别。本研究的结果不仅可用于北美水貂的鉴别,也为其他小型食肉动物的鉴别提供经验。     

Effects of dietary supplementation of nucleotides from late gestation to lactation on the performance and oxidative stress status of sows and their offspring

Increased metabolic burdens in breeding sows, which are induced by elevated systemic oxidative stress, could increase the need for nucleotides to repair lymphocyte DNA damage; however, de novo synthesis of nucleotides may be insufficient to cover this increased need. This study investigated the effects of dietary nucleotides on milk composition, oxidative stress status, and the reproductive and lactational performance of sows. Forty multiparous sows were assigned to 2 dietary treatments (Control group, and 1 g/kg Nucleotides group) based on a randomized complete block design using their BW at 85 d of gestation as a block. Sows from 2 groups were fed a restricted diet during gestation and ad libitum during lactation. The experiment lasted from 85 d of gestation to 21 d of lactation. The reproductive performance of sows and the growth performance of suckling piglets were measured. Oxidative stress parameters and milk components were also analysed. Data were analyzed using contrasts in the MIXED procedure of SAS. Sows in the Nucleotides group consumed more feed during the first week (P < 0.01) and from 1 to 21 d (P < 0.05) of lactation than those in Control group. Correspondingly, the litter weight gain of piglets showed a tendency to increase from cross-fostering to 9 d (P = 0.09) and from cross-fostering to 20 d (P = 0.10) in the Nucleotides group relative to the Control group. Additionally, the Nucleotides group was higher (P < 0.01) than the Control group in the concentrations of uridine 5''monophosphate, guanosine 5''monophosphate, inosine 5''monophosphate, adenosine 5''monophosphate and total nucleotides in milk. Furthermore, the Nucleotides group was higher (P < 0.01) than the Control group in the serum levels of total antioxidant capacity (P < 0.01) for sows at 109 d of gestation and glutathione peroxidase for weaning piglets, but lower at the levels of thiobarbituric acid-reactive substances (P < 0.05) in serum of weaning piglets. This study indicated that maternal dietary nucleotides could promote piglet growth, probably due to the higher lactational feed intake and higher concentration of nucleotides in the milk of sows, and lower oxidative stress for both sows and piglets.     

六株猪圆环病毒2型国内分离株的全基因组序列测定与分析

利用PK15细胞从临床发病猪和死亡猪淋巴结中分离到6株猪圆环病毒2型(PCV2)，对其全基因组序列进行了测定和分析。结果表明，除深圳分离株(SZ)的基因组全长为1768nt外，其余5株均为1767nt；6个分离株之间全序列核苷酸同源性为98．08％；与欧、美毒株之间同源性高，介于94．4％～99．7％。     

Effects of Cultivars,Transplanting Patterns,Environment, and Their Interactions on Grain Quality of Japonica Rice

With the increasing scarcity of rural labor, the rice transplanting pattern is encountering a shift from artificial transplanting (AT) to mechanical transplanting (MT) in numerous rice‐growing districts of China. The shift of transplanting patterns combined with altered growing environment during the grain‐filling stage in different years presumably affects rice quality. Nevertheless, related information is currently limited. This study investigated the effects of cultivars, transplanting patterns, environment, and their interactions on appearance, milling, eating, and nutritional qualities of four japonica rice varieties. The significant interactive effects of cultivars, environment, and transplanting patterns on almost all rice quality parameters (except Thr, Met, and Ile) were observed. Cultivars and environment were the main factors influencing rice appearance and milling and eating qualities. Cultivar was the primary factor affecting rice nutritional quality. Among all treatments, environment showed the strongest effect on percentage of chalky kernel, milled rice yield, peak viscosity, breakdown, setback, consistence, amylose, Glu, Tyr, and Met contents. However, Leu and Phe contents were unaffected by environment but only by cultivars and transplanting patterns. In addition to amylose and protein, Glu and Met contents were also involved in determining rice eating quality. Amino acid contents (except Cys, Tyr, and Met) were significantly negatively correlated with head rice yield, showing the function of amino acids in controlling rice milling quality. Percentage of chalky kernel as well as protein and almost all amino acid contents were significantly negatively correlated with the difference of maximum and minimum temperature (DMMT) and positively correlated with relative humidity (RH), whereas head rice yield did it reversely. Amylose content and setback were significantly negatively related to daily maximum temperature (DMAT), daily minimum temperature (DMIT), daily average temperature (DAT), and effective temperature accumulation (ETA). However, peak viscosity, breakdown, and consistence had contrary performances. According to these results, we can infer that DMMT and RH are important environmental factors affecting rice appearance, milling qualities, and nutritional qualities and that DMAT, DMIT, DAT, and ETA are key environmental factors influencing rice eating quality.     

深松整地机械化作业技术研究

论述深松整地机械化技术在辽宁省的实施效果及主要作用,从土壤条件、作业时间、作业深度、作业机具4个方面,介绍相关技术的应用要点,为该技术在全省的推广应用提供技术参考。     

寒地包衣玉米种子压缩特性试验研究

以寒地包衣玉米种子龙丹47为研究对象,对其物料特性进行了测量,得到三轴尺寸长9.287mm、宽7.64 mm、厚5.9 8 mm,休止角3 2.2 0°,自流角2 5.4 0°,千粒质量3 6 9.5 g。利用CTM2 0 5 0微机控制电子万能材料试验机,得到龙丹47玉米种子在平放、立放、侧放条件下的压缩特性曲线,经拟合曲线符合PEAL-READ模型,并获得了最大压缩载荷(平放379.5N、立放124.3N、侧放161.926N)、弹性模量(平放161.528 MPa、立放45.66MPa、侧放1 0 2.8 9 3 MPa)和抗压强度(平放1 0.8 4 4 MPa,立放3.5 5 2 MPa、侧放4.6 2 6 MPa)。结果显示:籽粒平放时的压缩载荷明显高于侧放和立放时的载荷,且平放压缩载荷是立放压缩载荷的2.6倍,是侧放压缩载荷的2.7倍;而侧放与立放的压缩载荷差异不大,表明籽粒在侧放和立放时更易受压缩外力的影响。