6个桑树品种的耐盐性鉴定试验

种植耐盐性强的桑树品种是土壤盐渍化地区生态修复的技术途径之一.选择桂桑优12、桂桑优62、抗青283×抗青10、桂桑5号、桂桑6号、桑特优2号共6个桑树品种的实生苗供试,将供试桑苗种植于质量浓度分别为0 g/L(CK)、2 g/L、4 g/L、6 g/L的NaCl溶液中,在历时28 d的盐胁迫试验期,每隔7d测定和统计各处理组桑苗的枝高、叶片数量、最大叶长、最大叶宽、受害叶片数量和枯死植株数量等,初步评价供试桑品种实生苗对盐胁迫的耐受能力.结果表明:在不同浓度盐胁迫处理组桑苗生长均较好的桑树品种是桂桑优12和桑特优2号;桑苗受盐害指数和死亡率最低的桑树品种均是桂桑优12.试验结果提示,6个供试桑树品种中桂桑优12是耐盐性最强的品种.     

1株库蠓源版纳病毒的分离与全基因组序列分析

为分析云南省虫媒病毒的种类与遗传特征,在云南省师宗县采集库蠓进行病毒的分离与鉴定;通过全长cDNA扩增与高通量测序技术获取病毒全基因组序列,进行序列比对与系统发生树构建。结果显示,从采集的库蠓样本中分离出1株可在C6/36细胞上引起细胞病变的毒株（YNSZ043）,病毒基因组为分节段双链RNA,琼脂糖凝胶电泳呈"2-4-3"的带型特征;电镜观察可见直径为70~80 nm,呈"指环状",表面具有纤维突起的病毒粒子。全基因组测序结果显示,YNSZ043毒株为版纳病毒（Banna virus,BAV）,基因组大小为20 683 bp,由Seg-1（3 762 bp）至Seg-12（861 bp）12个基因节段组成,与中国BAV毒株各基因节段的核苷酸序列相似性在64.8%~99.6%之间,氨基酸序列相似性在58.8%~100%之间,在系统发生树上YNSZ043毒株与中国分离的BAV聚为一簇,形成独立的中国进化支系。对决定BAV基因型的Seg-12分析结果显示,YNSZ043毒株属于A2基因型,该毒株的Seg-5/VP5与越南分离BAV毒株的核苷酸和氨基酸序列相似性高达97.1%和97.6%,表明该毒株的Seg-5基因节段很可能与越南毒株之间发生了基因重配。研究结果丰富了中国BAV的基因组序列,为开展云南省BAV的流行病学研究提供了参考。     

猪戊型肝炎病毒ORF3蛋白影响HepG2细胞核黄素代谢的lncRNA-mRNA调控网络的鉴定

为初步鉴定并挖掘出猪戊型肝炎病毒(Hepatitis E virus,HEV)ORF3蛋白影响HepG2细胞核黄素代谢信号通路的lncRNA-mRNA调控网络,本试验通过构建腺病毒过表达载体,制备高滴度过表达腺病毒,介导猪 HEV-ORF3在HepG2细胞中实现过表达。Western blotting检测ORF3蛋白过表达成功后,运用lncRNA高通量组学测序,筛选出差异表达的lncRNA并进行靶向差异基因预测,对lncRNA靶向差异基因进行GO功能和KEGG通路富集分析,初步鉴定出与核黄素代谢信号通路相关的lncRNA-mRNA调控网络。Western blotting结果显示,在约为12 ku处出现目的条带,说明成功实现腺病毒介导猪HEV-ORF3在HepG2细胞中过表达。lncRNA高通量组学测序结果显示,共发现102个显著差异表达lncRNAs表达量上调,80个显著差异表达lncRNAs表达量下调。GO功能和KEGG通路富集分析显示,初步挖掘出lncRNA(MSTRG.13995.2)和lncRNA(MSTRG.1960.1)可能是与核黄素代谢信号通路相关的显著差异表达lncRNA,分别通过顺式调控其靶向基因APC-5和FLAD1来影响核黄素代谢信号通路。本试验初步鉴定出lncRNA(MSTRG.13995.2)-APC5和lncRNA(MSTRG.1960.1)-FLAD1可能是影响HepG2细胞核黄素代谢信号通路的lncRNA-mRNA调控网络。     

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。     

珠江三角洲地区生鲜农产品冷链网络化运输技术

文章主要是对我国的珠江三角洲地区的生鲜农产品冷链的物流方式进行探究,结合现在我国的生鲜农产品冷链运输技术的现状和发展前景进行大胆的预测,并提出我国的珠江三角洲地区的生鲜农产品冷链的运输技术的网络化的建议。     

基于HHT变换的PSO-LSSVM耦合模型估算气象资料短缺地区ET_0

为准确估算气象资料短缺地区参考作物腾发量,构建了一种基于HHT变换的PSO-LSSVM耦合模型,并利用新疆和田气象站2000—2009年单日数据做训练、双日数据做验证。结果表明,该模型估算ET0方法明显优于常规的PSO-LSSVM和GRNN,预测精度较二者分别提高了15.7%~85.6%和15.8%~93.7%;该方法预测ET0的气象要素重要性为RsTmaxTminRHWn,利用该方法对气象要素组合为Tmax/Tmin/RH/Wn、Tmax/RH/Wn、Tmin/Wn、Wn条件下的ET0预测,MSE分别为0.407、0.185、0.149、0.135,说明该方法可以很好地估算资料缺失地区ET0。     

Study on Expression Difference of IGFBP-5 Gene in Different Tissues of Kazakh and Yanqi Horses

The study aimed to explore the mRNA expression pattern of insulin-like growth factor binding protein-5 (IGFBP-5) gene in different tissues of Kazakh and Yanqi horses.The expression of IGFBP-5 gene in different tissues of heart,liver,spleen,lung,kidney,small intestine,large intestine,cecum,intercostal muscles,longissimus dorsi muscles,brachialis muscle and gluteus in two horses were detected by Real-time quantitative PCR and compared the mRNA expression in the same tissues of two breeds.The results showed that the expression of IGFBP-5 in longissimus dorsi muscle and brachialis muscle of two breeds were significantly higher than other tissues including heart,liver,spleen,lung,kidney,small intestine,large intestine and cecum (P<0.05),and was the lowest in large intestine.The expression of IGFBP-5 in kidney,small intestine,large intestine,cecum,longissimus dorsi muscle and brachialis muscles of Yanqi horse were higher than in the same part of Kazakh horse,and among those in longissimus dorsi muscle and large intestine of Yanqi horse were extremely significantly higher than in the same part of Kazakh horse (P<0.01),and in small intestine and cecum of Yanqi horse were significantly higher than in the same part of Kazakh horse (P<0.05).The test was for further researching the biological function of IGFBP-5 gene,and it could provide a theoretical basis for genetic improvement of production performance of horse in our country.     

Effect of Vitamin E on Bovine Granulosa Cells Apoptosis and Proliferation through Cx43

The aim of this study was to determine the effect of vitamin E on Cx43,the mechanism and function of vitamin E on bovine granulosa cells apoptosis and proliferation.In this study,granulosa cells were isolated from bovine ovary and cultivated in vitro by adding different concentration of vitamin E (0,25,50,100,200 and 500 μmol/L) for 24 h.After cultured,apoptotic cells were detected by FCM,mRNA expression levels of BCL2/BAX、P53 and Cx43 genes were determined by Real-time PCR and cell proliferation was detected by CCK8.The results showed that compared to control group,100 μmol/L vitamin E could significantly inhibit the apoptosis of granulosa cells (P<0.05).Real-time PCR detection results showed that vitamin E significantly changed the mRNA expression levels of BCL2/BAX,P53,Cx43 genes (P<0.05).Vitamin E could significantly improve granulosa cells proliferation when granulosa cells were treated for 24 and 36 h (P<0.05).The results provided a theoretical basis on further analysis for studing the influence mechanism of vitamin E on oocytes development and maturity,and improvement of female animal reproduction by influencing granulosa cells proliferation and apoptosis.     

影响‘小凤兰’根状茎增殖、分化和壮苗的因素研究

以‘小凤兰’根状茎为试材,研究了影响‘小凤兰’根状茎增殖、分化、生根壮苗的因素。结果表明:MS基本培养基、蔗糖30g/L、6-BA 2.0mg/L+NAA 0.5mg/L、活性炭0.5g/L有利于根状茎增殖,而添加20%椰汁和番茄汁40g/L会抑制根状茎增殖。1/2MS基本培养基、蔗糖30g/L、6-BA 2.0mg/L+NAA 0.2mg/L、20%椰汁、1 000lx光照强度有利于根状茎分化,不加活性炭有利于芽分化,但抑制根分化,而添加活性炭则抑制芽分化,促进苗分化。添加土豆泥60g/L显著促进试管苗生长,3~4cm高的苗在培养基1/2MS+6-BA 0.1mg/L+NAA0.5mg/L+蔗糖20g/L+土豆泥60g/L+卡拉粉8g/L+活性炭0.5g/L中培养40d即可移栽。     

三种地被菊种子萌发及幼苗生长的化感效应比较

以天人菊、金鸡菊和蛇鞭菊为试材,比较两两混种与单种在种子萌发和幼苗生长阶段相互间的化感效应强度。结果表明:两两混种下,金鸡菊、蛇鞭菊对天人菊生长起促进作用,化感效应强度为金鸡菊蛇鞭菊;天人菊、蛇鞭菊对金鸡菊生长起抑制作用,化感效应强度为天人菊蛇鞭菊;金鸡菊、天人菊对蛇鞭菊生长起抑制作用,化感效应强度为金鸡菊天人菊。