刺参响应灿烂弧菌侵染的基因组DNA甲基化水平和转录组差异及其关联分析

为探讨病原菌胁迫下刺参(Apostichopus japonicus)基因组DNA甲基化水平和转录组表达的差异,本研究采用人工攻毒侵染胁迫,获得刺参化皮体壁组织,并以未攻毒组的健康体壁组织为对照,对刺参2种体壁组织进行全基因组甲基化测序(WGBS)和转录组高通量测序,解析刺参体壁基因组DNA甲基化差异,筛选响应病原胁迫的差异甲基化区域和差异表达基因。同时,通过基因组甲基化和转录组联合分析,筛选负相关关联基因,为解析刺参响应灿烂弧菌(Vibrio splendidus)侵染的分子机理提供基础数据。结果显示,刺参对照组和侵染组基因组总甲基化水平分别为(3.59±0.04)%和(3.87±0.27)%,侵染组甲基化水平显著升高;在发生甲基化的位点中,攻毒组和对照组的mCpG占比分别为83.06%和81.91%,mCpG为最主要的甲基化形式。本研究共筛选出差异甲基化区域(DMRs) 626 677个,注释到23 706个功能基因。转录组测序共检测到29 290个基因,筛选到差异表达基因496个,其中,上调基因214个,下调基因282个。在基因组甲基化与转录组的关联分析中,筛选到180个负相关关联基因,其中,差异甲基化区域位于启动子区域的负相关基因为60个。对负相关关联基因的GO和KEGG富集分析,筛选到相关通路和LRR、hsp20和CARD等关键基因。本研究将为解析刺参抗病的表观遗传调控机制提供数据,也为刺参抗病性状选育提供科学参考。     

大体积混凝土施工中的温度监测及裂缝控制

结合中联水泥集团南阳分公司水泥熟料生产线二期工程大体积混凝土的施工,提出在施工中防止大体积混凝土因水泥水化热引起的温度差而产生温度应力裂缝的措施。通过严格控制混凝土温度,降低内外温差,预防收缩缝,减少坍落度损失,延缓凝结时间等,保证大体积混凝土顺利施工。     

亲和素磁珠分离毛竹SSR标记方法的优化

在参考其它文献报道的基础上,构建毛竹SSR位点富集文库。用EcoRⅠ分别和DraⅠ、EcoRⅤ、SmaⅠ、PvuⅡ四种平端限制性内切酶对毛竹基因组DNA进行双酶切,酶切片段回收后与根据抑制性PCR原理设计的接头连接。在利用亲和素磁珠富集含有SSR序列的DNA片段过程中,对杂交、漂洗及洗脱步骤进行优化。通过三引物特异PCR筛选含有SSR序列的阳性克隆并结合测序结果对富集SSR文库进行评价,确定了一条简单、经济、高效分离竹类植物SSR标记的方法。     

Interactions between phosphorus availability and microbes in a wheat–maize double cropping system: A reduced fertilization scheme

Mechanisms controlling phosphorus(P) availability and the roles of microorganisms in the efficient utilization of soil P in the wheat–maize double cropping system are poorly understood. In the present study, we conducted a pot experiment for four consecutive wheat–maize seasons(2016–2018) using calcareous soils with high(30.36 mg kg^–1) and low(9.78 mg kg^–1) initial Olsen-P content to evaluate the effects of conventional P fertilizer application to both wheat and maize(Pwm) ...     

Identification of a conservative site in the African swine fever virus p54 protein and its preliminary application in a serological assay

BackgroundASF was first reported in Kenya in 1910 in 1921. In China, ASF spread to 31 provinces including Henan and Jiangsu within six months after it was first reported on August 3, 2018. The epidemic almost affected the whole China, causing direct economic losses of tens of billions of yuan. Cause great loss to our pig industry. As ELISA is cheap and easy to operate, OIE regards it as the preferred serological method for ASF detection. P54 protein has good antigenicity and is an ideal antigen for detection.ObjectiveTo identify a conservative site in the African swine fever virus (ASFV) p54 protein and perform a Cloth-enzyme-linked immunosorbent assay (ELISA) for detecting the ASFV antibody in order to reduce risks posed by using the live virus in diagnostic assays.MethodWe used bioinformatics methods to predict the antigen epitope of the ASFV p54 protein in combination with the antigenic index and artificially synthesized the predicted antigen epitope peptides. Using ASFV-positive serum and specific monoclonal antibodies (mAbs), we performed indirect ELISA and blocking ELISA to verify the immunological properties of the predicted epitope polypeptide.ResultsThe results of our prediction revealed that the possible antigen epitope regions were A23–29, A36–45, A72–94, A114–120, A124–130, and A137–150. The indirect ELISA showed that the peptides A23–29, A36–45, A72–94, A114–120, and A137–150 have good antigenicity. Moreover, the A36–45 polypeptide can react specifically with the mAb secreted by hybridoma cells, and its binding site contains a minimum number of essential amino acids in the sequence 37DIQFINPY44.ConclusionsOur study confirmed a conservative antigenic site in the ASFV p54 protein and its amino acid sequence. A competitive ELISA method for detecting ASFV antibodies was established based on recombinant p54 and matching mAb. Moreover, testing the protein sequence alignment verified that the method can theoretically detect antibodies produced by pigs affected by nearly all ASFVs worldwide.     

新发展格局视域下红色旅游高质量发展研究

将红色文化和旅游相融合,形成红色旅游是新时代旅游业升级转型之路。红色旅游对政治、经济、社会和文化有着较高的价值和效益,但存在品牌效应弱、基础设施不完备、同质化现象严重、服务和管理水平规范化不足等问题。在分析红色旅游内在价值和发展现状及困境的基础上,提出构建新型“红色旅游+乡村振兴”发展模式、沉浸式旅游体验、挖掘红色精神时代价值和推进人才队伍建设等建议,以期为红色旅游的健康发展提供政策支持。     

广东主要茶区春季杂草群落调查与区域差异分析

探明广东茶园春季杂草基本情况,为茶园杂草防控提供基础数据.2020年3-5月,在广东省粤东、粤西、粤北3个茶叶主产区31个茶园共620块样方开展茶园杂草群落调查,采用七级目测法,记录样方杂草种类及生长优势等级.调查共发现杂草313种,出现频度高于10％的杂草有126种,其中菊科杂草23种、禾本科12种、茜草科8种、蔷薇...     

发展有机蔬菜是我国蔬菜生产的必由之路

近 5 0年来石油工业迅速发展 ,在农业生产上由于超量使用化肥、农药 ,环境污染日益严重。因此 ,可持续农业发展 ,引起社会上的普遍关注 ,不使用化学合成的肥料、农药 ,改用有机肥、堆肥进行蔬菜生产是行之有效的方法。科学研究证明植物可以直接吸收有机物 ,用有机方法生产蔬菜 ,对环境无污染 ,产品质量好 ,味道纯正 ,有益健康。目前无土栽培大多数是全部用化肥配制营养液进行作物生产 ,产品硝酸盐含量偏高 ,对人体健康不利 ,如改用有机肥和生物防治病虫 ,就可以生产绿色食品蔬菜 ,这是 2 1世纪农业发展的方向     

花椰菜未受精子房、花托、花序轴离体培养诱导成苗研究

以花椰菜的花托、花序轴和未受了房为外植体,诱导出不定芽,并进行继代增殖培养,在工增殖培养基上每隔２５～３０天,不定芽可以增殖５～６倍,不加激素的１／２ＭＳ培养基上,试管苗生根率达到９８％以上,移栽成活率达到９５％以上。经观察,花托和未受精了再生植株是由外植体直接分化而来,不经过愈伤组织阶段,这样可避免不良变异,保持材料的遗传稳定性。     

Effects of oxidative stress and inflammatory response on kidney injury in mice with hyperthyroidism

AIM To explore the effects of oxidative stress and inflammatory response on kidney injury induced by hyperthyroidism in mice. METHODS Forty male Kunming mice were randomly divided into control group （n=20） and L-thyroxine （T4） group （n=20）. The mice in T4 group were intraperitoneally injected with T4 diluent at a dose of 1 mg/kg to induce hyperthyroidism, and those in control group were injected with normal saline of the same volume. After 7 weeks, the mice were weighed and dissected, the kidneys were removed and weighed, and the length of tibia was also measured. The activity of superoxide dismutase （SOD） and the content of malondialdehyde （MDA） in the kidney tissues were detected. The pathological changes of the kidney tissues were observed by HE staining. The levels of 4-hydroxynonenal （4-HNE）-modified proteins, interleukin-1 receptor-associated kinase 1 （IRAK1） and tumor necrosis factor receptor-related factor 6 （TRAF6） were determined by Western blot and immunohistochemistry. RESULTS Compared with control group, the body weight of the mice was decreased, while the kidney size and weight were increased significantly in T4 group. In addition, the ratios of kidney weight/body weight and kidney weight/tibia length were also increased （P<0.05）. In T4 group, the renal tubules were enlarged, and the epithelial cells of renal tubules were swollen and exfoliated, with vacuolar degeneration. Furthermore, reduced SOD activity, and increased MDA content and 4-HNE-modified proteins were found in T4 group, all of which were related to oxidative stress （P<0.05）. The levels of inflammation-related proteins IRAK1 and TRAF6 were significantly increased in T4 group （P<0.05）. CONCLUSION Excessive T4 may lead to kidney hypertrophy and injury in mice, and the mechanism may be related to oxidative stress and inflammatory response.