Toll样受体4(TLR4)基因剔除小鼠构建及初步表型分析

目的 利用CRISPR/Cas9技术构建Toll样受体4(TLR4)基因敲除小鼠模型,并观察突变小鼠对革兰氏阴性细菌脂多糖(LPS)刺激响应的变化。方法 针对TLR4基因外显子2设计并合成1对sgRNA片段,与编码Cas9的mRNA混合后通过受精卵显微注射方法,建立TLR4基因敲除小鼠,通过繁育获得基因敲除纯合子小鼠(TLR4^-/-小鼠);通过LPS刺激,分析TLR4^-/-小鼠对炎症应激的反应情况,并在分子和病理水平上和野生型对照(WT)进行比较。结果 PCR及测序检测表明TLR4基因外显子2在小鼠基因中被成功敲除;给予LPS刺激后,IL1β、IL6、MyD88、iNOS及TNFa等炎症因子的表达在野生型小鼠的心、肝和肺组织中显著上调,而在TLR4^-/-小鼠中则几乎没有变化;血生化指标显示LPS刺激后WT小鼠血清中的尿素(Urea)和肌酐(Cre)水平显著升高,而TLR4^-/-小鼠刺激前后无显著变化,病理分析同样发现TLR4^-/-小鼠能够抵抗LPS对肾组织的损伤。结论 利用CRISPR/Cas9技术成功构建了TLR4基因剔除小鼠模型,TLR4的缺失能够降低IL1β、IL6、MyD88、iNOS及TNFa炎症因子对LPS刺激的响应,抑制LPS引起的炎症反应及对组织的损伤。     

基于移动终端的稻田飞虱调查方法

【目的】建立一种基于移动终端的稻田飞虱调查方法,以减轻测报人员劳动强度,提高稻田飞虱调查的客观性,实现稻飞虱调查结果可追溯。【方法】利用Android相机、可伸缩手持杆和装载控制相机APP的Android手机研制了稻田飞虱图像采集仪。在Android开发环境下,利用socket通信和视频编码等技术,实现Android相机的视频采集与编码模块、视频传输模块和相机命令控制模块等。利用Android NDK开发和Java web等技术,实现手机端的视频预览模块、手机控制模块、图像上传模块等。相机实时拍摄的视频将压缩成H.264格式,通过RTSP/RTP协议控制其传输至手机端。手机端通过解压缩,实现实时预览相机所拍摄的视频,并控制相机拍摄水稻茎基部飞虱图像,同时将图像传输到手机端。稻飞虱识别算法部署在云服务器上。手机端可选择稻飞虱图像上传至云服务器,云服务器运行稻飞虱自动识别算法,结果返回至手机端。【结果】基于移动终端的稻田飞虱调查方法利用手机可以实时预览相机拍摄的水稻茎基部飞虱画面,控制相机拍照。云服务器上稻飞虱自动识别算法对图像中的飞虱平均检测率为86.9%,虚警率为11.2%;对稻飞虱各虫态平均检测率为81.7%,虚警率为16.6%。【结论】基于移动终端的稻田飞虱调查方法可以便捷地采集到水稻茎基部飞虱图像,实现稻田飞虱不同虫态的识别与计数。该方法可大大减轻测报人员的劳动量,避免稻飞虱田间调查的主观性,实现稻飞虱田间调查的可追溯。     

Increased microbial functional diversity under long-term organic and integrated fertilization in a paddy soil

Microbes play key roles in diverse biogeochemical processes including nutrient cycling. However, responses of soil microbial community and functional genes to long-term integrated fertilization (chemical combined with organic fertilization) remain unclear. Here, we used pyrosequencing and a microarray-based GeoChip to explore the shifts of microbial community and functional genes in a paddy soil which received over 21-year fertilization with various regimes, including control (no fertilizer), rice straw (R), rice straw plus chemical fertilizer nitrogen (NR), N and phosphorus (NPR), NP and potassium (NPKR), and reduced rice straw plus reduced NPK (L-NPKR). Significant shifts of the overall soil bacterial composition only occurred in the NPKR and L-NPKR treatments, with enrichment of certain groups including Bradyrhizobiaceae and Rhodospirillaceae families that benefit higher productivity. All fertilization treatments significantly altered the soil microbial functional structure with increased diversity and abundances of genes for carbon and nitrogen cycling, in which NPKR and L-NPKR exhibited the strongest effect, while R exhibited the least. Functional gene structure and abundance were significantly correlated with corresponding soil enzymatic activities and rice yield, respectively, suggesting that the structural shift of the microbial functional community under fertilization might promote soil nutrient turnover and thereby affect yield. Overall, this study indicates that the combined application of rice straw and balanced chemical fertilizers was more pronounced in shifting the bacterial composition and improving the functional diversity toward higher productivity, providing a microbial point of view on applying a cost-effective integrated fertilization regime with rice straw plus reduced chemical fertilizers for sustainable nutrient management.

     

外源施钙对盐胁迫下花生营养元素吸收与分配的影响

为解决盐碱地花生养分吸收不畅及分配受阻等问题,研究外源施钙对盐胁迫下花生氮、磷、钾、钙、镁吸收积累、分配特性和产量的影响,为盐碱地花生生产合理、高效施肥提供理论依据.以‘花育25号’为材料,在0.3%盐胁迫浓度下,设置4个Ca浓度梯度[T₁(0)、T₂(75)、T₃(150)和T₄(225) kg·hm^-2 CaO]进行盆栽试验.结果表明: 花生植株内养分含量依次为氮>钾>钙>磷>镁,苗期植株对氮和钙素的吸收中心均在叶片,磷、钾、镁的吸收中心为茎,苗期近一半的营养积累分配在各元素相应的生长中心.成熟期氮、磷、钾吸收中心转移到荚果中,尤以氮、磷在籽仁中的积累量居多,达72.3%～78.9%;钙、镁的吸收中心仍为叶片和茎,其分配比例分别为49.8%、32.6%.盐胁迫明显抑制花生植株各器官对氮、磷、钾、钙和镁各元素的吸收积累与分配,尤以对叶片和籽仁中氮素积累的抑制较为显著,但盐胁迫对荚果中镁的积累有促进作用.外源钙对盐胁迫下花生植株各器官氮、磷、钙和镁的吸收累积有明显的促进作用,尤其对籽仁中磷素积累的调节最为显著,其在籽仁中的积累量提高50%以上.适宜的钙施用量可显著促进盐胁迫下花生养分吸收积累量,提高花生成熟期荚果中氮、磷、钾的分配比,最终提高产量.综合各养分吸收、积累分配和产量结果,在0.3%盐胁迫条件下钙肥适宜施用量为150 kg·hm^-2 CaO.     

皖北地区土壤中不同形态氟含量及其影响因素

以皖北地区广泛分布的潮土、砂姜黑土、水稻土等为材料,采用连续化学提取的方法,研究了土壤中不同形态氟的含量、土体分布以及土壤氟形态与土壤理化性质的关系·结果表明:皖北地区土壤全氟含量为265·8～612·8mg·kg-1,平均含量为423·7mg·kg-1,土壤全氟含量高低排序为菜园土>潮土>水稻土>砂姜黑土>黄棕壤,菜地土壤存在氟积累现象.土壤氟主要以残余态氟形式存在,占土壤全氟的95%以上;其次为水溶态氟,占土壤全氟的1·5%左右;有机态氟、铁锰氧化物态氟和交换态氟含量均较少.28个土壤样本的水溶态氟含量范围为1·35～17·98mg·kg-1,平均含量为6·62mg·kg-1,含量较低的是硅质黄棕壤,含量较高的是菜地土壤、潮土和砂姜黑土.土壤水溶态氟含量与土壤pH和土壤有效磷含量呈极显著正相关,与土壤全磷含量呈显著正相关;土壤交换态氟与土壤物理性粘粒和小于0·001mm土粒含量呈极显著正相关;铁锰结合态氟与土壤全磷含量呈显著正相关;有机态氟与土壤有机质含量呈极显著正相关.对土壤氟形态与土壤性质的逐步回归表明,土壤pH,有效磷、有机质、全磷和物理性粘粒含量是影响土壤氟形态分布的主要因素,其中,土壤pH的影响最大.土壤水溶态氟含量与土壤母质有关,在浅湖沼相沉积物上形成的土壤水溶态氟含量最高,平均含量为9·05mg·kg-1;其次为在黄河冲积物上形成的土壤,平均含量为8·12mg·kg-1;在晚更新世黄土母质上形成的土壤水溶态氟平均含量为2·97mg·kg-1;在淮河冲积物上形成的土壤和轻质黄棕壤水溶态氟含量分别为2·05和1·91mg·kg-1.在土体中,土壤有机态氟和铁锰氧化物态氟含量随土层深度增加而减少,菜地土壤上部土层的全氟和水溶态氟含量均高于底土.     

不同生育期花生渗透调节物质含量和抗氧化酶活性对土壤水分的响应

通过防雨棚池栽试验,以不同花生品种为试材,研究了不同生育时期非充分灌溉对花生品种各生育期叶片膜脂过氧化、渗透调节物质含量和抗氧化酶活性的影响.结果表明,苗期和花针期灌水,叶片抗氧化酶活性、渗透调节物质和丙二醛(MDA)含量均有不同程度的降低.随生育期推进和土壤水分降低,其活性升高,但升幅因品种、抗氧化酶和渗透调节物质类型有异,两品种叶片超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、可溶性糖(SS)、可溶性蛋白质(Pr)、游离氨基酸(AA)和脯氨酸(Pro)含量均以对水分最为敏感的花针期升幅较大,且花育27号的SOD、CAT、Pr和AA的升幅大于花育20号;结荚期灌水后,各抗氧化酶和渗透调节物质未表现降低.两品种全生育期灌水处理与苗期灌水处理间的抗氧化酶活性、渗透调节物质和MDA含量差异均不显著.水分胁迫初期,抗氧化酶活性升高,但随胁迫时间延长其活性明显降低;而渗透调节物质和MDA含量显著高于各生育期灌水处理.POD活性变化对灌水处理响应较弱,SOD和CAT是花生适应土壤水逆境的主要保护酶.灌水处理对花生叶片抗氧化及渗透调节能力表现为花针期＞结荚期＞苗期,各渗透物质调节能力依次表现为脯氨酸、可溶性蛋白质＞可溶性糖＞游离氨基酸.     

Positive selection driving the evolution of a gene of male reproduction, Acp26Aa, of Drosophila: II. Divergence versus polymorphism

The evolution of the gene for a male ejaculatory protein, Acp26Aa, has been shown to be driven by positive selection when nonsibling species in the Drosophila melanogaster subgroup are compared. To know if selection has been operating in the recent past and to understand the details of its dynamics, we obtained DNA sequences of Acp26Aa and the nearby Acp26Ab gene from 39 D. melanogaster chromosomes. Together with the 10 published sequences, we analyzed 49 sequences from five populations in four continents. The southern African population is somewhat differentiated from all other populations, but its nucleotide diversity is lower at these two loci. We find the following results for Acp26Aa: (1) The R: S (replacement : silent changes) ratio is significantly higher in the between-species comparisons than in the within-species data by the McDonald and Kreitman test. Positive selection is probably responsible for the excess of amino acid replacements between species. (2) However, within-species nucleotide diversity is high. Neither the Tajima test nor the Fu and Li test indicates a reduction in nucleotide diversity due to positive selection in the recent past. (3) The newly derived nucleotides in D. melanogaster are at high frequency significantly more often than predicted by the neutral equilibrium. Since the nearby Acp26Ab gene does not show these patterns, these observations cannot be attributed to the characteristics of this chromosomal region. We suggest that positive selection is active, but may be weak, for each amino acid change in the Acp26Aa gene.      

内蒙古呼伦贝尔沙地不同樟子松林竞争强度的比较

利用全林木定位和单木竞争指数模型, 以受到林火干扰的樟子松林为例,分析了呼伦贝尔沙地樟子松林的竞争强度.结果表明：同一样地中,死亡林木的竞争强度均比较接近;活立木、全部林木中,伴生树种的竞争强度是樟子松的2～3倍;樟子松的竞争压力主要来自于种内,伴生树种的竞争则主要来自于种间.林火干扰与无林火（对照）样地间的比较表明,林火干扰样地樟子松活立木的竞争强度均显著小于对照样地;活立木竞争强度与其胸径间均服从幂函数关系(CI=AD^-B).地表火干扰可显著降低林木个体间的竞争强度,有利于存活林木个体的生长发育和大径阶林木的培育.