水稻BWMK1互作基因的分离

为探明BWMK1介导的信号传导途径,采用酵母双杂交系统,构建水稻稻瘟病菌诱导后的cDNA文库,以BWMK1为诱饵,对文库进行筛选,通过验证、测序和基因编码分析,获得了7个BWMK1互作基因BWIP1～BWIP7,分别位于水稻基因组第12、2、3、10、6、4、5染色体上；7个互作基因中,除BWlP2和BWIP7未找到同...     

新化水酒辣蓼草酒药工艺技术研究

以大米为原料，在酒药制作过程中添加辣蓼草，以糖化力和发酵力为评价指标，通过单因素试验和正交试验，探讨了辣蓼草添加量、种曲添加量、培曲时间对新化水酒酒药品质的影响，筛选出新化水酒辣蓼草酒药的最佳制作工艺条件。结果表明，新化水酒辣蓼草酒药的最佳制作工艺条件为：辣蓼草添加量6%，种曲添加量5%，培曲时间60 h，在此工艺条件下制得的酒药糖化力和发酵力分别为1 443 U/g和53.4 g/100 g。     

传统山西老陈醋增香酵母菌的筛选及其特性研究

为选育山西老陈醋用微生物发酵剂,从传统发酵工艺生产的山西老陈醋中分离筛选到5株增香酵母菌。结合形态学和酵母菌26S rDNA Dl/D2区序列分析,初步鉴定Y1为酿酒酵母(Saccharomyces cerevisiae),Y26为毕赤酵母(Pichia scaptomyzae),Y30为假丝酵母(Candida humilis),Y68为红酵母(Rhodotorula sp.),Y70为盔形毕赤酵母(Pichia membranifaciens)。采用固相微萃取和气-质联用技术分析这5株酵母发酵液的主要挥发性成分。结果表明,醇类和酯类挥发性成分种类和组成差异较大;对这5株酵母菌进行耐酸性、乙醇耐受性、高温耐受性发酵试验,结果表明毕赤酵母Y26具有优良的耐酸性、乙醇耐受性。综合其具有高产乙酯类、低产高级醇的特性,毕赤酵母Y26有望开发为山西老陈醋的新型发酵剂。     

月季bHLH基因的克隆、表达及其与MYB和WD40的互作分析

以月季‘月月红’(Rosa chinensis‘Slater’s Crimson China’)花瓣为材料,利用同源克隆法鉴定到1个Rcb HLH基因的c DNA全长序列,并研究了其生物信息学特征、组织特异性表达及其与MYB和WD40的互作关系。序列分析表明,Rcb HLH的开放阅读框长2 112 bp,编码703个氨基酸,基因登录号为KY783912。结构域序列分析显示,Rcb HLH为一个保守的N端含有碱性氨基酸的DNA结合区、C端含有α螺旋–环–α螺旋的b HLH蛋白。对Rcb HLH进行系统进化分析发现,Rcb HLH与草莓及其他蔷薇科植物的b HLH蛋白具有较高的同源性。组织特异性实时定量RT-PCR分析揭示Rcb HLH基因主要在花瓣中表达。酵母双杂交结果显示,Rcb HLH能与Pav MYB和Pav WD40相互作用形成MYB-b HLH-WD40复合物。以上结果为进一步研究Rcb HLH的功能鉴定奠定了基础。     

酵母浸膏对北柴胡种子萌发的影响及评价

以北柴胡种子为试验材料,分别以0.5％、1.0％、1.5％和2.0％的酵母浸膏液对北柴胡种子进行浸种,在种子萌发过程中测定相关的生理生化指标,探讨酵母浸膏液对北柴胡种子萌发的影响.含水量的测定采用烘干减重法,体积的测定采用排水法,可溶性糖含量的测定采用蒽酮比色法,蛋白质含量的测定采用考马斯亮蓝-G 250染色法,氨基酸含量的测定采用茚三酮显色法;利用隶属函数法对各项指标进行综合评价.结果表明:北柴胡种子在萌发过程中,种子内含水量的变化和其体积的变化呈一定的相关性,都是呈先下降后上升再缓慢增加的趋势;可溶性糖含量的变化呈先下降后上升再下降的趋势;蛋白质含量整体呈先急剧下降后上升再缓慢下降的趋势;氨基酸含量整体呈先上升后下降的趋势.综合各项指标进行综合评价,酵母浸膏液浸种对北柴胡种子的萌发具有明显的促进作用,其中以1.0％酵母浸膏液浸种效果最佳.     

Effects of dietary Bacillus coagulans and yeast hydrolysate supplementation on growth performance,immune response and intestinal barrier function in weaned piglets

The present study investigated the effects of Bacillus coagulans and yeast hydrolysate supplementation on growth performance, immune response and intestinal barrier function of weaned piglets. Twenty-four weaned piglets with an average body weight (BW) of 6.89 ± 0.15 kg were divided into four diets for 28 days. The treatments were basal diet (control), basal diet supplemented with antibiotic (20 mg/kg colistin sulphate and 40 mg/kg bacitracin zinc, AT), probiotics (400 mg/kg Bacillus coagulans ≥5 × 109 CFU/g, BC) or yeast hydrolysate (5000 mg/kg yeast hydrolysate, YH). Average daily gain (ADG) and average daily feed intake (ADFI) were improved by AT and YH diets (p < 0.05), while BC diet only increased ADG (p < 0.05). The complement 3 (C3), lysozyme (LZM) and tumour necrosis factor-α (TNF-α) concentrations in serum were increased in BC diet (p < 0.05). Feeding AT and YH caused the increase of jejunal villus height (p < 0.05), and a higher ratio of villus height/crypt depth was observed in AT, BC and YH groups (p < 0.05). The mRNA expression of zonula occludens-1 (ZO-1) in jejunal mucosa was up-regulated by AT, BC and YH diets (p < 0.05). Dietary AT, BC or YH inclusion decreased the interleukin-1β (IL-1β) concentration and TNF-α mRNA expression (p < 0.05), and YH supplementation even down-regulated toll-like receptor 4 (TLR4) and CD14 expressions (p < 0.05). In summary, the dietary administration of BC or YH both improves growth performance through promoting the intestinal barrier function, indicating both of them can serve as potential alternatives to antibiotics growth promoters for the piglet production.     

Construction and Identification of Yeast Surface Display Libraries of Ovis aries DRA Gene Exon 2

The specific primers were designed according to Ovis aries DRA gene sequence deposited in GenBank and the multiple cloning site of the plasmid pYD1,which was a vector used for protein surface display on Saccharomyces cerevisiae.The gene encoding DRA was amplified by PCR using the genomic RNA of Ovis aries.The 762 bp fragment was cloned and released in GenBank and registration number was KR422362.The PCR product was inserted into the yeast surface display plasmid vector pYD1 by double enzyme digestion.It was indicated that DRA gene was successfully integrated into the genome.Dot mutation was made at both ends of exon 2 in DRA gene for making restriction enzyme cutting site and design the exon 2 specific primers according to mutated Ovis aries DRA gene sequence.Sequenced exon 2 amplification products based on DNA pooling of sheep large sample template was analyzed the polymorphic loci.The polymorphic exon 2 246 bp fragment was obtained by double enzyme digestion and connected to surface display restructuring mutation carriers pYD1-DRA by the same double enzyme digestion,and then we successfully constructed yeast surface display libraries.We transformed it into Saccharomyces cerevisiae EBY100 cell.Yeast monoclone was identified by PCR amplification and sequencing,and we confirmed that DRA gene had been integrated into Saccharomyces cerevisiae genome.After galactose induced,it was detected that DRA gene library had been successfully demonstrated on the yeast cell surface under the fluorescence microscope by immunofluorescence method.     

葡萄VvSUC12基因启动子的克隆及上游调控因子鉴定

枝干病害葡萄溃疡病近年来严重限制了葡萄产业发展。研究表明葡萄蔗糖转运蛋白参与寄主植物和病原菌的互作过程。为解析蔗糖转运蛋白VvSUC12在葡萄免疫反应过程中的功能,本研究克隆了VvSUC12基因翻译起始位点上游1 500 bp的启动子区,通过生物信息学分析发现该区域包含4个Dof转录因子结合序列(A/TAAAG)及多种激素调节与防御相关的顺式元件。实时荧光定量分析显示,接种可可毛色二孢菌显著诱导VvSUC12和VvDof19基因的表达。通过酵母单杂交实验筛选得到与VvSUC12启动子互作的转录因子VvDof19。酵母双杂交实验证实VvDof19具有自激活活性;进一步通过烟草瞬时表达发现Dof19-GFP的相对GUS活性约为对照GFP的9倍,表明转录因子VvDof19能够激活VvSUC12基因的表达。研究结果为深入研究蔗糖转运蛋白在葡萄免疫反应中的功能奠定基础。     

净化SO_2气体的固定化微生物性能研究

利用SO2气体对某污水处理厂氧化沟采集的微生物进行诱导驯化,得到以氧化亚铁硫杆菌为优势菌的脱硫菌,研究了该固定化微生物脱硫菌在不同环境条件下的性能。结果表明,固定化后的微生物对热、pH等的稳定性提高,对金属等有毒物质的毒性抵抗力增强。     

酵母细胞壁多糖与铝硅酸盐复合物对猪生长性能、免疫指标及养分消化率的影响

本研究旨在探讨酵母细胞壁多糖(yeast polysaccharide,YP)与铝硅酸盐复合物(alumi-nosilicate complex,AC)对猪的生长性能、免疫指标及养分消化率的影响。试验选用平均体重为(66.18±1.26)kg的"杜×长×大"三元杂交猪140头,随机分成5组,每组4个重复,每个重复7头猪。各组分别饲喂添加不同剂量复合物的试验饲粮,A组为对照组,不添加复合物;B组添加0.1%YP;C组添加0.1%YP和0.9%AC;D组添加0.1%YP和1.9%AC;E组添加0.1%YP和2.9%AC。试验期51 d。结果表明:1)D、E组平均日增重显著低于对照组(P<0.05),各组平均日采食量和料重比差异不显著(P>0.05);2)与对照组相比,D组血清谷胱甘肽过氧化物酶(GSH-Px)活性及补体3(C3)、免疫球蛋白M(IgM)、免疫球蛋白A(IgA)、免疫球蛋白G(IgG)水平显著提高(P<0.05),血清超氧化物歧化酶(SOD)活性和补体4(C4)水平无显著差异(P>0.05);3)各组养分消化率无显著差异(P>0.05)。由此可见,YP和AC复合使用降低了育肥猪的生长性能,0.1%YP和1.9%AC同时添加可增强生长猪的免疫功能。