野生大豆内生真菌对作物促生作用的研究

为了实现绿色生态农业、农作提质增效,加快生防菌剂的研发进度。本研究利用具有广谱拮抗玉米大斑病和大豆疫霉病野生大豆内生真菌Y6R15和Y2S2为试材,通过菌液灌根和叶喷处理,分析不同处理下玉米和大豆苗期干物质和根系指标的变化。结果表明：拮抗菌株Y6R15对玉米叶片和根干重促生作用明显,均提高根长度、根表面积、根体积、根尖数等指标,以根冠比指标标准Y6R15叶喷效果为最佳方式,依据根系指标特征变化Y6R15灌根效果好于叶喷。拮抗菌株Y2S2和Y6R15处理均对大豆有促生作用,效果显著,Y6R15菌株对玉米和大豆均有促生作用。     

Improvement,identification, and target prediction for miRNAs in the porcine genome by using massive,public high-throughput sequencing data

Despite the broad variety of available microRNA (miRNA) research tools and methods, their application to the identification, annotation, and target prediction of miRNAs in nonmodel organisms is still limited. In this study, we collected nearly all public sRNA-seq data to improve the annotation for known miRNAs and identify novel miRNAs that have not been annotated in pigs (Sus scrofa). We newly annotated 210 mature sequences in known miRNAs and found that 43 of the known miRNA precursors were problematic due to redundant/missing annotations or incorrect sequences. We also predicted 811 novel miRNAs with high confidence, which was twice the current number of known miRNAs for pigs in miRBase. In addition, we proposed a correlation-based strategy to predict target genes for miRNAs by using a large amount of sRNA-seq and RNA-seq data. We found that the correlation-based strategy provided additional evidence of expression compared with traditional target prediction methods. The correlation-based strategy also identified the regulatory pairs that were controlled by nonbinding sites with a particular pattern, which provided abundant complementarity for studying the mechanism of miRNAs that regulate gene expression. In summary, our study improved the annotation of known miRNAs, identified a large number of novel miRNAs, and predicted target genes for all pig miRNAs by using massive public data. This large data-based strategy is also applicable for other nonmodel organisms with incomplete annotation information.     

纳米孔测序技术在疾病检测中的研究进展

近几年兴起的纳米孔测序技术(nanopore sequencing)为新一代测序技术,又称第三代测序技术,已开始应用于细菌、病毒、抗生素耐药及动物疫病等多个领域的检测。虽然在实际应用中仍存在一些瓶颈,包括测序精度不高、样品制备过程需要优化等,但该技术在疾病检测方面存在明显优势,包括可直接检测碱基修饰,超长读长、高分辨率和实时测序以及强大的生物信息学支持。未来,随着准确性的提高和性能的不断改进,加之在成本、测序时间和生物信息学分析方面的明显优势,纳米孔测序技术有望在临床实验室成为疾病检测的重要选择。本文通过综述该技术的原理、应用及在疾病检测中的优势,以期为纳米孔测序技术的进一步应用研究提供支持。     

猪GRP78基因克隆、生物信息学分析及组织表达谱研究

本研究旨在克隆猪葡萄糖调节蛋白78（glucose-regulated protein 78，GRP78）基因，并进行生物信息学分析，探讨其在猪不同组织中的表达情况。根据GenBank上公布的猪GRP78基因序列（登录号：XM_001927795.6）设计引物，PCR扩增及测序获得猪GRP78基因CDS序列，使用在线软件分析GRP78的理化性质、跨膜结构、信号肽、疏水性、保守结构域、二级结构和三级结构，并进行同源性比对及系统进化树构建，利用实时荧光定量PCR方法检测猪GRP78基因在各组织中的表达情况。结果显示，猪GRP78基因CDS区长1 965 bp，可编码654个氨基酸。生物信息学分析表明，GRP78理论分子质量为72.3 ku，等电点（pI）为5.06，半衰期为30 h，其水溶液在280 nm处的消光系数为30 495，肽链N端为蛋氨酸（Met），不稳定系数为32.39，属于稳定蛋白。脂肪系数为85.40，总平均疏水指数为-0.496，无跨膜结构，存在信号肽序列，说明该蛋白属于分泌型蛋白；GRP78蛋白只包含1个超家族保守结构域：HSP70结构域。蛋白二级结构分析显示，GRP78蛋白中α-螺旋、延伸链、β-转角和无规则卷曲分别为40.06%、20.49%、8.10%和31.35%。同源性比对结果显示，猪GRP78基因与人（登录号：NM_005347.4）、小鼠（登录号：NM_001163434.1）、大鼠（登录号：NM_013083.2）、山羊（登录号：XM_005687138.3）、牛（登录号：NM_001075148.1）核苷酸序列的同源性分别为93%、91%、90%、95%、95%，氨基酸序列的同源性分别为99%、98%、98%、99%、99%，各个物种之间GRP78基因保守性较高。实时荧光定量PCR结果显示，GRP78基因在心脏、肝脏、脾脏、肺脏、肾脏、小肠、胃、卵巢、输卵管、乳腺、小脑、大脑、垂体等组织中均有表达，在心脏、脾脏、肺脏中相对高表达。本研究结果为今后深入研究GRP78基因的生物学功能提供了基础材料。     

政策认知、执行力度与资源化利用满意度——基于河北省380家生猪养殖主体调研数据(上)

参与主体对于政策的评价是衡量政策实施效果的重要指标，探明政策参与主体满意度影响因素有助于了解主体自身需求，从而提高政策实施水平。文章基于对河北省380家生猪养殖主体调研数据，运用多元有序Logistic回归方法，分析生猪养殖主体自身政策认知以及相关政策执行力度对畜禽养殖主体废弃物资源化利用满意度的影响。结果表明：养殖主体资源化利用政策了解程度能够显著提升其资源化利用政策满意度；仅考虑养殖主体政策参与力度，随着参与力度增加，满意度亦会显著提升。但综合考虑其他因素，参与力度对于满意度提升效果则不再显著；养殖主体感知政策执行力度增加不利于政策满意度提升。此外，生猪养殖主体废弃物资源化利用政策了解程度增加，可以显著提升其报告资源化利用政策“满意”可能性。而单纯提高资源化利用政策执行力度并不能显著提高养殖主体资源化利用政策满意度。因此，要提高养殖主体政策满意度，应多渠道提升宣传力度，鼓励养殖主体积极参与，提高其参与意识，不断改善现有政策执行方式。     

优质棉中棉所127与高衣分材料杂交分离群体纤维产量和品质表型评价及典型相关分析

以纤维品质优异的陆地棉品种中棉所127为父本，以高衣分品系GH07-44为母本，构建了F₂和F_2:3分离群体，并对F₂和F_2:3群体的产量和纤维品质性状进行表型评价及典型相关分析。结果发现，分离群体各性状存在丰富的遗传变异和超亲分离，变异系数为1.15%～15.19%；F₂群体中铃重的变异系数最大，2个世代中纤维成熟度指数的变异系数均最小。超高亲比例为1.71%～66.86%，其中F_2:3群体马克隆值的超高亲比例最高，2个世代中最小的超高亲比例性状均为上半部平均长度。简单相关分析表明：衣分与马克_{隆值、成熟度指数呈极显著正相关，与上半部平均长度、长度整齐度指数呈极显著负相关；铃重与马克隆值呈极显著正相关，与F}2群体的成熟度指数呈极显著正相关，与F_2:3群体的成熟度指数呈显著正相关。典型相关分析表明，产量性状组与纤维品质性状组间存在相关关系，主要表现在衣分与上半部平均长度呈负相关、与马克隆值呈正相关，随着世代的增加，产量和品质性状的相关性降低，因此实现产量和纤维品质同步改良虽有一定难度但有可能实现。本研究筛选出2个世代衣分均超过43%且纤维品质较好的材料9个（马克隆值均值＜4.5、上半部平均长度均值＞30 mm），为棉花产量和纤维品质的同步改良提供了基础材料。     

乡村振兴视域下山区农业发展路径——以皖南山区黄山市精致农业发展为例

以皖南山区黄山市为例对精致农业发展情况进行了分析,重点从3个方面阐述了其存在的问题和不足,之后从5个方面系统探讨在实施乡村振兴战略背景下提升山区精致农业发展的有效途径.     

An Electrothermochromic Fabric Prepared by Electrodeposition of Polypyrrole on Single Side

An electrothermochromic fabric triggered by electric voltages was prepared by combining the electric heating feature of conductive polypyrrole (PPy) and the temperature-response color-changing feature of thermochromic inks. PPy was deposited selectively on the carbon-coated side of the sailcloth fabric via electrochemical polymerization. Thermochromic inks were painted on the reverse side with white color. The obtained fabric could be heated to about 48 °C and change its color significantly under an applied voltage of 3 V.     

Isolation and Characterization of GhLEA3 Gene from Upland Cotton and Its Expression in Response to Low Temperature Stress

[Objective] Based on the analysis of cotton GhLEA3 gene structure and its expression model in chilling stress, we investigated the resistance function of GhLEA3 in response to cold stress. [Method] We cloned GhLEA3 from upland cotton by homologous sequence cloning way. We analyzed the properties of the GhLEA3, and constructed phylogenetic tree by bioinformatics analysis. We constructed the transient expression vector 35S∷GhLEA3-GFP by In-Fusion connection technology and studied the subcellular localization of GhLEA3. The expression of GhLEA3 gene in leaves of TM-1 at three-leaf stage under low temperature stress treatments (4 ℃, 24 h) was performed. Agrobacterium tumefaciens mediated floral dipping method was used to transform the gene with expression vector 35S∷GhLEA3-GFP into Arabidopsis thaliana wild type. Low temperature germination experiment at 4 ℃ was conducted to verify germination ability of the transgenic Arabidopsis thaliana T₃ seed. The transgenic Arabidopsis seedlings were treated at 4 ℃(low temperature), and all the leaves were taken to measure their electrical conductivity. [Result] The coding sequence of GhLEA3 gene is 1 218 bp, which encodes 405 amino acids. GhLEA3 protein includes 4 functional domains of PF02987. Phylogenetic analysis showed that the similarity of amino acid sequences between upland cotton GhLEA3 and Arabidopsis thaliana AtLEA3 was 52.6%. GhLEA3 might be mainly localized in vacuoles and small vesicles. GhLEA3 was up-regulated in leaves after low temperature treatment. The germination rate of transgenic Arabidopsis thaliana of T₃ generation at low temperature was significantly higher than that of wild type, and its electrical conductivity of leaves after low temperature treatment was significantly lower than that of wild type. [Conclusion] GhLEA3 belonged to the member of LEA3 family. Phylogenetic analysis showed that GhLEA3 had the closest relationship with AtLEA3. GhLEA3 was induced by low temperature stress and may play an important role in improving cold resistance.     

川西地区鸡滑液囊支原体分离株的部分生物学特性及其VlhA基因遗传进化分析

为了解鸡滑液囊支原体(MS)在川西地区的感染情况,本研究采集15个肉鸡场疑似MS感染的病鸡咽拭子、跗关节和胸部滑液囊样本共75份,对经PCR检测为阳性的样本进行MS分离,并对分离株的主要生物学特性进行研究,以及对分离株的VlhA基因进行遗传进化分析。结果显示,75份样本MS PCR阳性检出率为41.33%(31/75),11个鸡场感染该病,场阳性率为73.33%(11/15);但仅从其中3个鸡场分离到9株MS,分离株菌落与菌体形态与已知MS培养特性相符,各分离株培养浓度为10~4CCU/mL^10~6CCU/mL,分离株以5×10~5CCU接种7日龄SPF鸡胚,鸡胚于接种后7 d^9 d死亡,且分离回收到接种菌株;9株MS分离株VlhA基因的同源性为86.1%~99.9%,与参考株同源性为86.2%~95.4%,其中分离自同一鸡场的7株MS VlhA基因同源性为96.4%~99.9%;分离株VlhA基因遗传进化分析显示,其中分离自两个不同鸡场的两株MS与国内流行株的亲缘关系最近,而分离自另一鸡场的7株MS与中东地区分离的3株MS亲缘关系较近,表明MS在川西地区肉鸡场呈高感染率,MS VlhA基因变异较大。本研究为川西地区MS的进一步研究提供了基础材料和科学依据。