利用CRISPR/Cas9系统敲除葡萄中VviEDR2提高对白粉病的抗性

利用CRISPR/Cas9系统定点编辑葡萄白粉病感病基因VviEDR2(Enhanced disease resistance2),在VviEDR2的DUF1336结构域设计靶位点VviEDR2-T1,构建CRISPR/Cas9敲除载体,通过农杆菌介导法转化‘无核白’葡萄胚性愈伤组织。对PCR阳性植株进行靶位点扩增测序,结果表明,共有8个转基因植株在靶位点处发生不同类型的双等位基因突变,编辑效率为32%;突变体植株生长势较弱,叶片较小,茎秆丛生、细弱。进一步对突变体植株进行抗病检测,结果表明,接种葡萄白粉菌(Erysiphe necator Schw.)5d后,突变体植株叶片上白粉菌孢子仅能萌发出少量较短初级菌丝,表皮细胞产生大量明显的H2O2,而野生型叶片中白粉菌萌发出大量初级菌丝、次级菌丝和吸器,无明显H2O2产生。这些结果表明,可以利用CRISPR/Cas9技术编辑葡萄感病基因VviEDR2,提高葡萄白粉菌抗性。     

宁麦9号与扬麦158株高及其构成因素的遗传解析

宁麦9号与扬麦158是我国长江中下游麦区的主栽品种和骨干亲本,长江中下游麦区近3年来审定品种中80%都是其衍生后代,研究其性状的遗传具重要意义。以宁麦9号与扬麦158为亲本构建的包含282个家系的重组自交系群体为材料,利用Illumina 90k芯片对群体进行基因型分析,建立高密度遗传图谱。连续3个生长季对株高及节间长度、穗长等株高构成因素进行测定,结合遗传图谱对株高及相关性状进行QTL定位,获得14个控制株高及其构成因素的稳定表达位点。通过进一步位置比对,聚焦到6个染色体区段,初步明确了各节间对株高的遗传调控机制。同时,将6个染色体区段中同源性较低的连锁标记转化为适用于高通量筛选的KASP标记,利用101份区域试验材料进行标记效应验证,结果显示聚合Qph-2D与Qph-5A.1两个位点具有较高的选择效率,继续聚合Q2A后,中选材料显著减少,可能降低选择效率;对Q2A与Q5A两个一因多效位点的选择建议以降低株高的等位变异为主;Qd1-5D可作为穗下节间(D1)的选择标记对株高展开优化选择。期望以上结果能为长江中下游麦区的小麦株高遗传改良提供帮助。     

史氏鲟精子超微结构

利用扫描和透射电镜观察了史氏鲟精子形态和超微结构。精子具有顶体、头部、中段和单鞭毛等部分。顶体长0.99±0.06μm,宽0.87±0.09μm。细胞核从后往前逐渐变细,前端宽度为0.88±0.04μm,后端宽度为1.26±0.06μm,细胞核长7.29±0.32μm。核内含有3条核管(E),核管上行至顶体下行至植入窝而止。中段紧接头后部,长为0.51±0.12μm,宽0.91±0.05μm。中段含有线粒体、中心粒复合体和鞭毛的起始部分。线粒体分2~3层排列,线粒体中可见髓样嵴结构。在细胞核与中段接合部细胞核向内凹陷形成植入窝,纤维体位于植入窝内,其后是中心粒复合体。中段后缘延长为袖套,袖套腔中含有线粒体、脂质空泡。鞭毛从袖套中伸出,由轴丝组成,为典型的"9+2"微管结构。     

水稻不育突变株沈农05-9的形态和细胞学特征鉴定

以辽粳5号与丰锦杂交的高代系的突变株沈农05-9和其原始株系为材料,采用醋酸洋红染色压片技术及石蜡切片技术.进行了形态学特征、小孢子发生的细胞学特征观察.结果表明:该不育突变株花粉表现典败和染败,多数花粉母细胞减数分裂不形成二分体,而形成多核的花粉母细胞;花药表现干瘪且纤维化严重,绒毡层细胞在小孢子发育过程中提前解体.该不育突变株的减数分裂方式类似于萍乡型显性不育系及早稻昆植不育系.导致败育的直接原因为绒毡层细胞提前解体.     

H9亚型禽流感病毒的分离与鉴定

从临床表现呼吸困难并伴有啰音、肺部和支气管黄色干酪样栓塞为特征的病鸡肝脾肺混合匀浆中分离到1株病毒（YD株）,该病毒能凝集鸡红细胞并能被AIVH9标准阳性血清抑制,对热不稳定且对热敏感;对鸡胚和番鸭胚的最小致死量分别为10^-7和10^-6,其MDT分别为78h和70h;鸡胚尿囊液毒的HA效价和EID50明显高于番鸭胚尿囊液毒;能扩增出大小约为1027bp的特异性目的片段。结果表明．该分离毒为H9亚型禽流感病毒。     

小鼠FGF9在大肠杆菌中的表达与纯化

为了建立小鼠FGF9的原核表达体系,获得纯化的FGF9重组蛋白,采用PCR技术扩增FGF9,并将其克隆入原核表达载体pET30a(+)中,经测序验证后将该重组质粒转入大肠杆菌BL21(DE3)中进行表达,采用亲和层析技术进行目的蛋白的纯化,并用Western blotting进行了免疫原性验证.测序结果显示插入到载体中的片段与FGF9的天然序列一致,在BL21(DE3)中表达出了可溶性的FGF9重组蛋白,其表达量达到总蛋白的40 %;用镍螯合亲和层析方法获得了纯度大于95 %的FGF9重组蛋白.     

肉鸽H9亚型禽流感的诊断与病毒分离及鉴定

通过分子PCR检测,初步证明在湖北一发病肉鸽群感染H9亚型禽流感病毒。病原分离后,盲传3代,其血凝效价达到109~1011,经生物学特性进行测定,其鸡胚半数致死量为107.5,9日龄鸡胚平均致死时间为54.3 h,1日龄雏鸡脑内接种致病指数为1.8,6周龄小鸡静脉接种致病指数为2.0,证明该H9亚型禽流感为毒力较强。将该病毒回归非免疫肉鸽,能产生典型的H9亚型禽流感症状,采集其肺、喉头、泄殖腔等病变组织进行病毒分离,均可分离到H9亚型AIV。     

人工催熟日本鳗鲡精子的显微和超微结构

对经人工催产获得的日本鳗鲡（Anguilla japonica）精子的显微和超微结构进行了观察。光镜下观察,日本鳗鲡精子有两种形态结构,一种精子的细胞核为圆形或近 似圆形,这精子较小,细胞核长径为(2.57±0.62) μm,短径为(2.11±0.59) μm,鞭毛长度为(38.35±7.71) μm;另外一种精子的细胞核为“眉形”或“新月形”,精子较大,细胞核长径为(7.66±1.09) μm,短径为(2.54±0.46) μm,鞭毛长度为(38.35±9.02) μm。透射电镜观察结果显示：圆形精子头部的顶端无顶体,植入窝位于细胞核底端的中间,由细胞核向内凹陷而成,呈一沟状,其走向与精子的长轴平行。中心粒复合体位于植入窝内。细胞核的下端有2～3个线粒体。基体的头端呈筒状,由电子致密物构成。基体的尾端分裂成9束。尾部从袖套腔中伸出且细长,轴丝始端与基体的尾端相连,微管多呈电子致密状,轴丝为典型的“9+2”结构。圆形精子为日本鳗鲡正常成熟的精子。“眉形”或“新月形”精子头部细胞核弯曲,在弯曲处有一圆形的球状物,球状物内含有线粒体和中心粒复合体,轴丝为“9+0”结构。这种精子可能存在发育缺陷或没有达到完全正常成熟。     

Bird flu (H7N9) outbreak and its implications on the supply chain of poultry meat in China

During the past two decades, food safety issues in China not only posed serious threats to Chinese consumers but also damaged the image of Chinese products internationally. In China, food safety is not only about scientific discoveries, advanced laboratories, and sanitation equipment; it is more about the role of different players in the food supply chain. The poultry meat supply chain is instrumental in the spread of the avian influenza A virus (H7N9), raising questions about how policymakers respond to such threats and whether industries need to be restructured to manage and control this epidemic so that it does not recur. As a short-term measure, to prevent the spread of this disease, government authorities enforced the closure of live bird markets (LBM) in disease-affected areas of China. However, in the long term, the poultry meat supply chain needs to be restructured. The aim of the current study was to analyze distribution channels for chicken meat in China and then describe arrangements in poultry meat sectors that incorporate small- and medium-scale producers into the supply chain while responding to shifts in LBMs. We also assessed the role of LBMs in spreading H7N9 and how these interventions affect the poultry meat supply chain in the Chinese market.     

Reduction in cadmium accumulation in japonica rice grains by CRISPR/Cas9-mediated editing of OsNRAMP5

Cadmium (Cd) intake is harmful to human health and Cd contamination in rice grains represents a severe threat to those consuming rice as a staple food. Knockout of Cd transporters is a promising strategy to reduce Cd accumulation in rice grains. OsNRAMP5 is the major transporter for Cd and manganese (Mn) uptake in rice. Nevertheless, it is uncertain whether knockout of OsNRAMP5 is applicable to produce low Cd rice without affecting plant growth and grain yield. In this study, we adopted CRISPR/Cas9-based gene editing technology to knock out OsNRAMP5 in two japonica varieties. We generated three independent transgene-free osnramp5 mutants and investigated the effect of osnramp5 mutations on Cd accumulation and plant growth. Hydroponic experiments showed that plant growth and chlorophyll content were significantly reduced in osnramp5 mutants at low Mn conditions, and this defective growth in the mutants could be fully rescued by supply of high levels of Mn. Cd and Mn accumulation in both roots and shoots was markedly reduced in the mutants compared to that in wild-type plants. In paddy field experiments, although Cd in flag leaves and grains was greatly reduced in osnramp5 mutants, some agronomic traits including plant height, seed setting rate, and grain number per panicle were affected in the mutants, which ultimately caused a mild reduction in grain yield. The reduced plant growth in the mutants can be attributed to a marked decrease in Mn accumulation. Our results reveal that the manipulation of OsNRAMP5 should be treated with caution: When assessing the applicability of osnramp5 mutants, soil pH and soil water content in paddy fields need to be taken into consideration, since they might affect the levels of available Mn in the soil and consequently determine the effect of the mutation on grain yield.