Comparison of flagellin and an oil-emulsion adjuvant in inactivated Newcastle disease vaccine in stimulation of immunogenic parameters

The present study was designed to investigate the potential application of native (N) and recombinant (truncated modified [tmFliC] and full-length [flFliC]) flagellin proteins along with inactivated Newcastle disease virus (NDV). Fifty six SPF chickens were immunized twice with PBS (control), inactivated NDV (Ag), inactivated NDV/flFliC (AgF), inactivated NDV/tmFliC (AgT), inactivated NDV/N (AgN), commercial vaccine containing Montanide (Vac) and Vac/N (VacN), with a two-week interval. Blood was collected weekly and spleens were harvested after chickens were sacrificed. Interleukin-6 (IL-6) and tumor necrotic factor-α (TNF-α) gene expression in peripheral blood mononuclear cells were analyzed by Real-Time PCR. Antibody response was assessed by haemagglutination inhibition (HI). Cellular activity was quantified by MTT assay. Results showed that the most IL-6 and TNF-α gene expression was observed in AgF group (P < 0.01). The lowest gene expression among vaccinated groups was observed in Ag group for IL-6 and Ag and Vac group for TNF-α. The highest HI titer was observed in Vac, VacN, AgF and AgT groups. The AgF group showed the highest cellular activity (P < 0.01). In conclusion, flagellin-adjuvanted groups showed a pro-inflammatory effect and acted similarly to or better than the Vac group. Hence, flagellin can be proposed as a potential adjuvant for ND vaccine.     

黄瓜枯萎病新型抑制蛋白L37的研究

 从土壤中分离到对黄瓜枯萎病病原菌有抑制作用的枯草芽孢杆菌(Bacillus subtilis) , 发酵产物经DEAE - 纤维素、Sephadex G-100柱层析, 分离到15 kD抑菌蛋白L37, SDS-PAGE显示纯度达到电泳纯, N - 末端氨基酸序列同源性检测结果提示可能是一种新型的抑菌蛋白。蛋白质抑菌结果表明, 对多种病原真菌具有抑制效果, 且对蛋白酶、温度和大部分有机物不敏感, 是稳定的抑菌蛋白。对病原菌菌丝抑制机理研究表明, 抑菌蛋白具有严重扭曲、断裂、阻止生长等抗生效果。应用发酵蛋白产物进行两年大田防效试验表明, 对黄瓜枯萎病防效显著, 最高达到90% , 具有显著提高产量的作用。     

川西北高寒沙地不同年限高山柳林下优势植物碳、氮、磷生态化学计量特征

通过采集川西北高寒沙地不同年限(6、18、24年)高山柳林下3种优势植被藏沙蒿、裂叶独活和镰荚棘豆,分别测定分析3种植被叶片、根部C、N、P化学计量特征变化特征。结果表明:不同年限高山柳林下植被C、N、P含量及其比值间存在显著差异且呈现出不同变化趋势。林下植被C含量整体下降;叶N含量呈上升趋势,根N含量随年限增长而下降;除藏沙蒿外,林下植被P含量变化不显著;C∶N变化范围为1.92~12.86;C∶P为29.18~196.88;不同年限高山柳林下植被N∶P间虽存在差异,但均表现出主要受到P限制,表明该区域植被生长主要受P限制,应注意P养分的适当补充。     

四倍体鸭茅产量及其构成因素的QTL定位

产量相关性状是复杂的数量性状,对鸭茅的单株产量及构成因素进行QTL分析,可提高育种中对产量性状优良基因选择的效率,同时为鸭茅遗传改良、基因克隆及分子标记辅助育种提供理论依据。以四倍体鸭茅“楷模”和“01436”为亲本杂交而成的作图群体为试验材料,于2014年对洪雅、宝兴两个不同生境下鸭茅株高、旗叶长、倒二叶长、旗叶宽、倒二叶宽、茎粗、花序长、分蘖数、单株干重等9个产量相关性状进行了表型鉴定及相关性分析。此外,在已构建的高密度鸭茅分子遗传图谱的基础上,采用MapQTL 5.0进一步对这些性状QTL定位分析。结果表明,绝大多数性状在亲本间呈显著差异且都整体表现出连续变异,符合数量性状遗传的特征;相关性分析表明,大多数产量相关性状均与干重极显著正相关,与单株干重相关性最好的依次为分蘖、株高;QTL分析发现,控制该9个农艺性状的QTL共60个,洪雅38个,宝兴22个,这些QTL分别定位于分子连锁图谱的1、2、3、4、5共5个连锁群上,单个QTL的贡献率为5.7%~24.7%,单个性状QTL个数为2~15个。其中,控制株高、花序长的QTL各12个,控制倒二叶长、茎粗的QTL各4个,控制旗叶宽、倒二叶宽的QTL各2个,控制单株干重的QTL有6个,影响分蘖的QTL为3个,与旗叶长相关的QTL最多15个。     

Mechanisation of organic fertiliser spreading,choice of fertiliser and crop residue management as solutions for maize environmental impact mitigation

The environmental impact of crop production is mainly related to fossil fuels consumption and to fertilisers application. Emissions arising from the spreading of organic and mineral fertilisers are important contributors for impact categories such as eutrophication and acidification. The choice of the fertilisers and of the spreading techniques as well as the crop residues management can deeply affect the environmental impact related to crop cultivation.In this study, seven scenarios describing fertilising schemes characterised by different organic and mineral fertilisers and by different mechanisation were compared. The aim is to evaluate, using the Life Cycle Assessment (LCA) method, how the environmental performances of grain maize production were affected by these different fertilisers schemes. The study was carried out considering a cradle to farm gate perspective and 1 t grain maize was selected as functional unit. Inventory data were collected on a farm located in Po Valley (Northern Italy) during year 2013 and were processed using the composite method recommended by the International Reference Life Cycle Data System (ILCD). The compared scenarios involved organic and mineral fertiliser distribution and were: pig slurry incorporation after >3 days after spreading (BS), fast pig slurry incorporation within 2 h from spreading (AS1), direct soil injection of pig slurry (AS2), pig slurry incorporation (after >3 days) with straw collection (AS3), digestate spreading instead of pig slurry (after >3 days) (AS4), only mineral fertilisers (i.e. urea and superphosphate) distribution (AS5) and only mineral fertilisers (i.e. calcium ammonium nitrate and superphosphate) distribution (AS6).The results were not univocal, since climate and soil conditions as well as physical and chemical fertiliser characteristics differently affected the environmental load, especially for particulate matter formation, terrestrial acidification and terrestrial eutrophication impact categories. AS1 and AS2 showed the most beneficial results for these impact categories (between ↙67% and ↙73% respect to worst scenario). AS6, on the opposite, showed the highest environmental impact for those impact categories mainly affected by energy and fossil fuel consumption (climate change, ozone depletion, human toxicity with carcinogenic effect, particulate matter, freshwater eutrophication, freshwater ecotoxicity and mineral, fossil and renewable resources depletion), categories on which AS3 and AS4 were the best solutions. AS3 was the most impacting for terrestrial acidification and eutrophicationA sensitivity analysis was carried out varying grain maize yield (mostly affected: marine eutrophication) and ammonia volatilisation losses due to organic fertilisers (mainly affected: terrestrial acidification and eutrophication).The achieved results can be useful for the development of ⬓spreading rules⬽ that drive the application of organic fertilisers in agricultural areas where there is an intense livestock activity.     

三种融合标签对大熊猫轮状病毒结构蛋白VP6-VP7可溶性表达的影响

大熊猫轮状病毒(giant panda rotavirus,GPRV)是引起幼龄大熊猫腹泻的主要病原,对圈养大熊猫产生了较大的危害。轮状病毒结构蛋白VP6是一种载体蛋白,可介导黏膜免疫反应。VP7是轮状病毒结构蛋白中主要的中和抗原。因此,VP6-VP7的融合表达作为候选抗原对该病的防治具有重要的意义。传统大肠埃希菌原核表达存在表达量低、可溶性差以及纯度低等弊端。本研究使用醛缩酶(EDA)、谷胱甘肽S-转移酶(GST)、麦芽糖结合蛋白(MBP)3种融合标签,以实现获得表达量高和纯度高的GPRV-VP6-VP7重组表达蛋白。将扩增的VP6、VP7基因片段利用同源重组酶构建到含3种融合标签的表达载体pET21b上,将重组质粒转化至大肠埃希菌Rosetta(DE3)感受态细胞中进行低温诱导表达。用Ni-柱亲和层析法纯化目的蛋白,SDS-PAGE和Image J分析蛋白表达量和可溶性,Western blot分析得到表达的重组表达蛋白正确且具有蛋白活性。实验结果证明,EDA标签能显著促进VP6-VP7蛋白的原核可溶性表达,提高VP6-VP7蛋白表达量。     

小麦族的基因组显性及其育种学意义

小麦族包含大量由不同基因组组成的异源多倍体物种。同一个异源多倍体物种的不同基因组可能对表型性状产生非对称性贡献,例如小麦属多倍体物种的形态分类特性,更像其A基因组供体物种,这种现象称为A基因组显性。由于基因组显性,小麦族形成了以A、D、U、St为轴心(显性)基因组的异源多倍体物种簇。异源多倍体物种的基因组显性可能与其进化适应优势的形成有关。在育种方面,基因组显性影响多倍体新作物开发及小麦-外源染色体易位设计。     

水稻粒宽突变体gw87的基因定位及候选基因分析

【目的】对水稻粒宽突变体gw87（grain width 87）进行表型鉴定、遗传分析、基因定位及候选基因分析,为探明该基因调控水稻籽粒大小的分子机制及应用潜力奠定基础。【方法】利用甲基磺酸乙酯诱变籼稻恢复系材料676R,获得一份籽粒宽度和千粒重显著增加的突变体gw87。对该突变体进行表型观察、农艺性状调查及外源油菜素内酯（BL）敏感性、叶绿素含量、光合参数测定,了解其表型特征及生理特性。调查gw87与676R杂交F₁的表型和F₂群体的分离情况,分析其遗传行为;选取该群体中的突变植株进行高通量测序,并利用gw87与粳稻品种日本晴杂交的F₂代作为定位群体,通过MutMap分析和分子标记定位,遴选候选基因并进行DNA和cDNA测序验证。利用qRT-PCR分析gw87和676R中BR合成途径基因OsDWARF4、D11和D2的表达差异。【结果】与野生型亲本676R相比,gw87突变体的株高降低,节间缩短,其中,倒一节间长度缩短最大,并呈现出扭曲的形态;叶片长度减少,宽度增加;单株有效穗数、主穗穗长和结实率显著降低,但籽粒宽度和千粒重显著增大。BL敏感性试验显示,gw87突变体幼苗对外源BL的敏感性降低。光合色素和光合参数测定表明,gw87光合色素含量增加,光合速率也有所增加。遗传分析表明gw87的突变性状是由1对隐性核基因控制。MutMap分析显示gw87突变基因位于第5染色体中部,在该染色体区域仅有1个碱基突变引起编码氨基酸变化;分子标记连锁分析表明该突变基因位于InDel标记X2和X3之间约101 kb的染色体区域;综合这两方面分析结果,最后遴选出gw87候选基因是编码一个含有AP2/EREBP DNA绑定结构域的转录因子基因LOC_Os05g32270。对该候选基因进行DNA和cDNA测序验证,发现gw87突变体中该基因DNA的第1 041位的碱基由G突变为A,导致与该位点相邻的76 bp内含子序列被剪接为外显子,引起阅读框移码,蛋白翻译提前终止。qRT-PCR分析显示gw87突变体中BR合成途径基因的表达量显著上调,表明gw87突变体中BR信号减弱。【结论】gw87是smos1、shb、rla1、ngr5的新等位突变体,但与这些突变体表型不同,gw87籽粒的宽度和千粒重显著增加,可能是LOC_Os05g32270的突变位点不同,导致其编码蛋白的功能活性不同所造成。