长白猪Plin3、Plin5基因克隆及表达规律研究

脂滴包被蛋白3(Plin3)和脂滴包被蛋白5(Plin5)是细胞内脂滴包被蛋白(PAT)家族的成员,在脂滴合成方面具有重要作用。为探究Plin3和Plin5在长白猪中的序列和表达特征,利用PCR技术扩增该基因,采用生物信息学分析两者的序列特征,并利用实时荧光定量PCR技术检测其在长白猪11个不同组织中的表达特征。结果表明,长白猪Plin3序列全长1 403 bp,Plin5全长1 397 bp,两种蛋白二级结构均以α-螺旋为主,无规则卷曲次之,不存在跨膜结构,无信号肽结构,有多个磷酸化位点。检测两种基因在不同组织中的表达水平发现,Plin3在长白猪的11个组织中均有表达,其中在脾脏中的相对表达量显著高于其他组织(P<0.05),肝脏次之;Plin5在脂肪组织中的相对表达量显著高于其他组织(P<0.05),而在大肠和小肠中不表达。本研究为进一步探究长白猪Plin3和Plin5在脂质代谢中的作用机制提供了理论依据。     

基于基因表达式编程的作物水分生产函数构建

作物水分生产函数的确定是农业水资源优化配置的关键。该研究采用农业水文生态系统模型（Agro-Hydrological & Chemical and Crop systems simulator, AHC）与基因表达式编程（Gene Expression Programming, GEP）相结合的方法构建作物水分生产函数。以河套灌区3种主要作物（葵花、玉米、小麦）为研究对象,采用AHC模型模拟作物产量等,构建基于GEP算法的作物水分生产函数,探讨考虑盐分胁迫的作物水分生产函数构建的思路与方法。结果表明：1）作物模拟产量与地下水埋深、地下水矿化度和灌水量等因素有关。2）构建作物水分生产函数的最优输入因子组合为地下水埋深、灌溉量、蒸散发、地下水矿化度、土壤根层盐分对作物胁迫因子、土壤根层含水率。3）应用作物水分生产函数估算不同灌溉定额条件下作物产量（预测产量）,并与AHC模型计算的产量（模拟产量）进行比较,玉米、葵花、小麦预测产量与模拟产量具有很好一致性,其决定系数分别是0.96、0.93、0.96,平均相对误差均小于5%,满足计算精度要求。因此,该研究所构建的作物水分生产函数可以较准确地估算盐分胁迫下作物产量,为农业节水与灌溉水高效利用提供科学参考。     

基于两点机器学习方法的土壤有机质空间分布预测

准确预测土壤有机质（Soil Organic Matter,SOM）空间分布对精细农业、耕地质量建设、生态环境保护以及固碳减排等均具有重要的意义。该研究探讨了基于两点机器学习方法（Two-point Machine Learning,TPML）提高SOM空间分布预测的可行性。以黑龙江省海伦市为研究区,以气候、地形地貌、社会经济和空间位置信息等因素作为辅助变量,充分利用空间位置信息和属性相似关系,有效处理SOM空间分布异质性及其与辅助变量间关系异质性,以提高TPML方法进行SOM空间分布预测的精度。采用随机森林、基于随机森林的回归克里格、反距离权重法和普通克里格（Ordinary Kriging,OK）方法作为对比,以平均绝对误差（Mean Absolute Error,MAE）、均方根误差（Root Mean Square Error,RMSE）、预测值与真实值相关系数（r）和决定系数（R2）作为评价指标,进行不同样本量下的多组对比试验,评价不同方法的预测精度。结果表明：1）研究区SOM含量在1.775～7.188 g/kg之间,平均值为3.179 g/kg,空间分布不均匀,呈东高西低的分布趋势。2）在不同样本量条件下,与其他模型相比,TPML的预测精度均最高,其MAE（0.088～0.097 g/kg）和RMSE（0.116～0.139 g/kg）均为最小,r（0.992～0.996）和R2（0.971～0.985）均为最高。3）预测值的误差标准差（理论误差）与实际误差具有相似的空间模式,说明TPML可以为预测结果提供合理的不确定性估计。综上,TPML模型可以通过同时利用空间自相关性和属性相似性来提高预测精度,该模型适用于预测具有一定空间自相关性且具有可用辅助数据的资源环境变量。     

基于全卷积神经网络的猪背膘厚快速准确测定

为了有效地弥补猪B超图像人工手绘背膘厚的不足,为生猪育种工作提供更精准和稳定的背膘厚测定新方法。该研究将全卷积神经网络（Fully Convolutional Networks,FCN）模型应用于猪B超图像的背膘分割和背膘厚测定上,开发出一套使用Python调用FCN模型对猪B超图像背膘厚进行自动测定的系统。通过开展验证集验证试验、屠宰比对试验和人员比对试验,发现模型测定结果和标注结果之间差异不显著（P>0.05）,两者相关系数达0.92（P<0.01）;B超标准测定背膘厚和FCN分割测定背膘厚的相关系数达到0.97（P<0.01）;专家组组内标准差为0.17（最小）,行业外组组内标准差为1.67（最大）,而FCN分割结果稳定性强,不受人员因素的影响。因此,该方法可以实现对外种猪B超背膘厚的精准、快速、稳定测量,减少猪场对专业人员的依赖,降低测定人员培训成本,减少工作人员工作量。     

Soil texture affects the conversion factor of electrical conductivity from 1:5 soil-water to saturated paste extracts

Electrical conductivity (EC) of soil-water extracts is commonly used to assess soil salinity. However, its conversion to the EC of saturated soil paste extracts (EC_e), the standard measure of soil salinity, is currently required for practical applications. Although many regression models can be used to obtain EC_e from the EC of soil-water extracts, the application of a site-specific model to different sites is not straightforward due to confounding soil factors such as soil texture. This study was conducted to develop a universal regression model to estimate a conversion factor (CF) for predicting EC_e from EC of soil-water extracts at a 1:5 ratio (EC_1:5), by employing a site-specific soil texture (i.e., sand content). A regression model, CF=8.910 5e^{0.010 6sand}/1.298 4 (r²=0.97, P < 0.001), was developed based on the results of coastal saline soil surveys (n=173) and laboratory experiments using artificial saline soils with different textures (n=6, sand content=10%-65%) and salinity levels (n=7, salinity=1-24 dS m^-1). Model performance was validated using an independent dataset and demonstrated that EC_e prediction using the developed model is more suitable for highly saline soils than for low saline soils. The feasibility of the regression model should be tested at other sites. Other soil factors affecting EC conversion factor also need to be explored to revise and improve the model through further studies.     

Methanotrophy-driven accumulation of organic carbon in four paddy soils of Bangladesh

Biological methane oxidation is a crucial process in the global carbon cycle that reduces methane emissions from paddy fields and natural wetlands into the atmosphere.However,soil organic carbon accumulation associated with microbial methane oxidation is poorly understood.Therefore,to investigate methane-derived carbon incorporation into soil organic matter,paddy soils originated from different parent materials(Inceptisol,Entisol,and Alfisol) were collected after rice harvesting from four major rice-producing regions in Bangladesh.Following microcosm incubation with 5%(volume/volume)¹³ CH₄,soil¹³ C-atom abundances significantly increased from background level of 1.08% to 1.88%–2.78%,leading to a net methane-derived accumulation of soil organic carbon ranging from 120 to 307 mg kg^-1.Approximately 23.6%–60.0% of the methane consumed was converted to soil organic carbon during microbial methane oxidation.The phylogeny of¹³ C-labeled pmoA(enconding the alpha subunit of the particulate methane monooxygenase) and 16 S rRNA genes further revealed that canonical α(type II) and γ(type I) Proteobacteria were active methane oxidizers.Members within the Methylobacter-and Methylosarcina-affiliated type Ia lineages dominated active methane-oxidizing communities that were responsible for the majority of methane-derived carbon accumulation in all three paddy soils,while Methylocystis-affiliated type IIa lineage was the key contributor in one paddy soil of Inceptisol origin.These results suggest that methanotroph-mediated synthesis of biomass plays an important role in soil organic matter accumulation.This study thus supports the concept that methanotrophs not only consume the greenhouse gas methane but also serve as a key biotic factor in maintaining soil fertility.     

Genomic characterization of multidrug-resistant extraintestinal pathogenic Escherichia coli isolated from grain culture soils

Multidrug-resistant (MDR) Escherichia coli, mainly extraintestinal pathogenic E. coli (ExPEC), has been widely reported in infections worldwide. In agricultural soils, manure is a hotspot for the dissemination of antimicrobial resistance genes (ARGs) and pathogenic bacteria; however, MDR bacteria have also been reported in soils with no history of manure use. In addition, cross-resistance and co-resistance have been described as responsible for the metal-driven selection of bacteria resistant to antimicrobials. Therefore, the aim of this study was to analyze three MDR E. coli isolates obtained from Brazilian grain culture soil samples with no history of manure use by whole-genome sequencing. The MDR E. coli isolates were recovered from soils from corn and coffee fields, and presented resistance to β-lactams, quinolones, aminoglycosides, tetracyclines, sulphonamides, and dihydrofolate reductase inhibitor. Resistome analysis showed ARGs to several antimicrobials (i.e., β-lactams, tetracyclines, aminoglycosides, sulphonamides, trimethoprim, phenicols, fosfomycin, and macrolides) as well as several metal resistance genes and antibacterial biocide resistance genes. In addition, known mutations in quinolone-resistance-determining regions of GyrA (Ser83Leu and Asp87Asn), ParE (Ser458Thr), and ParC (Ser80Ile) were also detected. Virulome analysis showed the presence of virulence genes (lpfA, mcmA, gad, mchF, iroN, cma, and iss) associated with ExPEC. Multidrug-resistant ExPEC isolates were assigned to phylogenetic group B1. The presence of MDR B1-ExPEC in soil samples shows the ability of these isolates to survive in soils. This study reports for the first time some sequence types (i.e., ST345, ST448, and ST1146) of MDR E. coli in Brazilian soils. Therefore, these findings contribute to the monitoring of antimicrobial resistance and surveillance studies based on whole-genome sequencing worldwide.     

钙信号抑制剂加剧低钾胁迫对烟草幼苗光合特性及钾吸收的影响

  【目的】  研究低钾胁迫抑制钙信号传导对植株幼苗叶片光合特性及钾吸收的影响,为缓解作物的低钾胁迫提供理论依据。  【方法】  以烟草品种K326为试验材料进行了室内水培试验。烟草幼苗先在营养液正常钾(K+ 5 mmol/L)和缺钾(K+ 0.15 mmol/L)营养液中生长8 天,然后在营养液中分别施加钙通道有机阻断剂Vp、钙通道无机阻断剂LaCl₃和钙离子螯合剂(EGTA)钙信号抑制剂,继续生长4天后取样,测定植株干鲜重、电导率、活性氧含量、叶片抗氧化酶活性、叶片光合特性等指标和植株地上、地下部钾素浓度及钾离子吸收相关基因表达量。  【结果】  1)不添加钙通道抑制剂或螯合剂条件下,低钾胁迫的烟株地上部和根系干重较常钾水平显著降低了59.29%、39.82%,鲜重显著降低了51.58%、48.19%,除水汽压饱和亏外的其余5项光合特性指标显著降低,植株地上部和根系钾含量分别显著降低了39.31%和77.05%,而叶中可溶性蛋白、相对电导率、过氧化氢含量则显著增加,NKT2、NtKC1、NtHAK1、NtHAK5基因的相对表达量分别增加了2.09、3.32、3.23、12.34 倍。2)常钾水平下,3个钙抑制剂处理未明显影响地上部和根系干鲜重,而低钾水平下,钙抑制处理的地上部鲜重和干重显著高于无钙抑制剂对照。两个钾水平下,与各自对照相比,3个钙抑制剂处理的叶片相对电导率、活性氧积累量均显著增加,抗氧化酶SOD和POD活性、叶绿素含量显著减少,光合特性指标(除水汽压饱和亏外)明显降低。3)两个钾水平下,3个钙信号抑制剂处理的烟株地上部和根系钾含量均显著低于其对照,低钾胁迫下平均降低幅度分别达到25.54%和53.36%。低钾胁迫下,钾离子通道基因NKT2、NtKC1和转运蛋白基因NtHAK1、NtHAK5在根系中的相对表达量分别平均降低了60.60%、50.62%、38.61%、69.79%,钙通道相关基因NtCNGC1和NtCNGC11在根中的相对表达量分别平均降低了95.36%和87.04%。  【结论】  低钾胁迫明显影响烟株幼苗生长、光合作用及对钾素的吸收和积累。低钾水平下,外施钙通道抑制剂或螯合剂可进一步降低烟草叶片抗氧化酶活性,减弱烟株叶片光合作用,并抑制根中钾吸收相关基因的表达和烟株对钾素的吸收。     

荔枝P型ATP酶基因家族鉴定及其在采后贮藏过程中响应拮抗菌N-1的表达模式分析

P型ATP酶是植物体内一种重要的膜转运蛋白,在果实能量代谢和酸代谢中发挥重要作用,并广泛参与植物离子运输、抗逆、细胞信号转导等各项生命活动,然而在荔枝或其他无患子科植物中尚未对P型ATP酶基因家族进行全面分析,本研究基于‘妃子笑’荔枝转录组数据,通过对这些基因进行生理生化分析、生物信息学分析和表达分析,共鉴定了28个P型ATP酶基因家族成员基因,探讨P型ATP酶在荔枝采后贮藏过程中的潜在功能。结构和系统发育分析表明该基因可以分为5个亚家族,同一亚族的基因在基因结构和motif基序方面具有很大的相似性,其蛋白二级结构主要以α-螺旋为主,亚细胞定位预测分析发现LcPAs多定位于线粒体（21/28）。通过转录组分析,在采后贮藏期间,荔枝P型ATP酶家族基因中上调表达的基因明显高于下调表达基因,暗示其可能在荔枝采后品质劣变过程中发挥着积极的抵御作用。使用拮抗菌N-1处理荔枝可以有效抑制其果实采后褐变和病害的发生,结合RT-qPCR结果,发现拮抗菌N-1处理能诱导LcPA1、LcPA7、LcPA8和LcPA19在整个贮藏过程中的上调表达,并维持了较高的ATP酶活性,进一步说明P型ATP酶家族基因在抑制荔枝病变过程中发挥着重要作用。本研究为了解荔枝P型ATP酶基因家族基因的进化和功能分析提供了参考,为深入了解拮抗菌N-1保鲜机理的分子调控机理提供新的证据。     

不同氮素水平下恢复系及杂交稻组合氮肥高效利用

为了比较利用MAS选育的氮高效型恢复系及其杂交稻组合在不同氮素水平下的农艺性状差异。本研究以抗病虫害和氮高效的多基因聚合恢复系R4302、R4312及其配制的8个杂交稻组合为试验材料,‘华占’‘天优华占’为对照,设置0 kg/hm²（N₀）、112.5 kg/hm²（N_112.5）、150 kg/hm²（N₁₅₀）、187.5 kg/hm²（N_187.5）4个氮肥水平,成熟期测定与分析供试材料的单株有效穗数、单株产量、干物质产量、氮肥农学利用率、氮肥偏生产力和收获指数。结果表明,不同氮素水平下R4302各农艺性状均优于‘华占’,R4312除氮肥农学利用率外其他农艺性状优于或接近‘华占’,表明OsTCP19基因的丢失并未影响R4302、R4312氮肥的高效利用;杂交稻组合天丰A/R4302、天丰A/R4312为氮高效型组合;贵A/R4302、科20A/R4312、香龙A/R4302、科4364A/R4312为低氮高效型组合;荃9311A/R4302属于双高效型组合;科20A/R4302属于高氮高效型组合;恢复系可用单株产量、氮肥农学利用效率、干物质产量其中1项指标或有效穗数来评估氮肥的利用效率;杂交稻组合可用单株产量、干物质产量、氮肥偏生产力、氮肥农学利用率、有效穗数、收获指数其中1项指标来评估氮肥的利用效率。本研究为通过氮高效基因选育氮肥高效利用型恢复系和杂交水稻组合提供参考。