甘薯小象甲是甘薯上危害最为严重的世界性害虫，对生态环境和社会经济遭受巨大损失。为提高甘薯小象甲综合防治的效果和深入理解其遗传进化机制，我们对甘薯小象甲功能基因组学进行了的深入研究。利用 Illumina 和 PacBio技术，对单对交配15代的甘薯小象甲进行测序。获得了甘薯小象甲成虫染色体水平的基因组，基因组大小为338.84Mb，Contig N50 和 Scaffold N50 分别为 14.97 Mb和34.23 Mb。预测重复序列为 157.51 Mb和 11907 个编码蛋白质基因。共有 337.06 Mb长度的基因组序列被定位到 11 条染色体上，其中能够确定顺序和方向的序列长度为 333.79 Mb，占定位到染色体上总序列长度的 99.03 %。比较基因组学分析表明，甘薯小象甲和中欧山松大小蠹亲缘关系较近，约 1.38 亿年前从中欧山松大小蠹的祖先分化而来。许多重要的基因家族在甘薯小象甲基因组中得到了扩张，包括农药解毒、耐冷应激和化学感觉系统相关基因家族。为了进一步解析气味结合蛋白在甘薯小象甲嗅觉识别过程中的作用，竞争性结合分析结果表明，CforOBP4-6对性信息素及其他配体具有很强的结合亲和力。高质量的甘薯小象甲基因组图谱为揭示其分子生态学基础、群体遗传和适应性进化机制及绿色有效防控的新方法和新技术提供了坚实的基础。

猪的呼吸道是多种病原微生物的定殖场所，包括常见的四种猪源支原体，分别是猪肺炎支原体（Mhp），猪鼻支原体（Mhr），猪絮状支原体（MF）和猪滑液支原体（MHS）。猪源支原体主要寄居于猪气管黏膜表面，最具代表性的Mhp是引起猪气喘病的主要病原，并易与其他猪呼吸道病原混合或继发感染，引发猪呼吸道疾病综合征，使得感染猪产生慢性持续性呼吸系统疾病，给养猪业带来巨大的经济损失。沿着猪呼吸道上皮细胞的纤毛的黏附是猪源支原体成功感染的前提。猪原代气管上皮细胞（PTEC）是研究包括猪源支原体的各种猪呼吸道病原发病机制的合适模型，但原代PTEC细胞短暂的寿命极大地限制了其应用前景。因此，构建永生化的猪气管上皮细胞，对包括Mhp在内的所有猪源支原体等猪呼吸道病原，具有较高的应用价值。

我们首先提供了详细的分离和培养原代PTEC的方法步骤。随后通过用含有人端粒酶逆转录酶（hTERT）的重组构建的质粒pEGFP-hTERT转染原代PTECs，通过两轮G418的抗性筛选，建立永生化的猪气管上皮细胞系（hTERT-PTECs），并可传代至60代以上。通过比较原代与永生化细胞的上皮细胞表面标志物角蛋白18的表达，细胞周期，细胞生长曲线，端粒酶活性，染色体核型分析，端粒酶基因的蛋白质印记检测，软琼脂和裸鼠成瘤试验，增殖能力等相关基因的定量PCR检测，不仅证实了永生化细胞具备了原代细胞的形态及功能特性，与原代PTECs相比，hTERT-PTEC也具有更长的寿命，更高的端粒酶活性和增殖活性。裸鼠体内未表现出恶性表型，表明该细胞系不具有致瘤性。将不同的猪源支原体菌株感染hTERT-PTECs，通过颜色变化单位CCU₅₀黏附率的定量计算，hTERT-PTECs对所有猪源支原体易感，且原代和永生化细胞之间的黏附能力无显著差异。而对于代表性的Mhp，DNA拷贝定量实时PCR测定，间接免疫荧光测定和蛋白质印迹分析表明hTERT-PTECs能够粘附不同毒力的Mhp菌株。总之，与原代PTECs类似，hTERT-PTECs可以广泛用作猪源支原体的粘附细胞模型，并可用于多种猪呼吸道病原体的感染研究。

哺乳动物子宫内膜的发育是一个涉及众多的调节因素的复杂过程。环状RNA （circRNAs）是一种自然存在的内源性非编码RNA家族的重要成员之一，有研究报道circRNAs在多种生理过程中发挥着重要的调控作用。目的：本研究旨在分析奶山羊发情周期非妊娠子宫内膜组织中circRNA的表达谱。方法：采用链特异性去核糖体转录组测序技术对发情第5天和发情第15天奶山羊的非妊娠子宫内膜组织样进行circRNAs表达谱分析，筛选两个时期差异表达的circRNAs并利用RT-qPCR进行验证；然后利用生物信息学分析两个时期差异表达circRNAs的宿主基因，进而进行GO 和KEGG分析；最后利用Targetscan 7.0和miRanda网站分析circRNAs和miRNAs的靶向结合关系。结果：奶山羊发情第5天和发情第15天的非妊娠子宫内膜组织中共有2331个差异表达的circRNAs（P<0.05），RT-qPCR检测的10个circRNAs变化趋势与测序结果一致；其中circRNA1460的宿主基因Nipped-B 样蛋白（Nipped-B-like，NIPBL）和circRNA8694的宿主基因钙反应性转录因子（calcium responsive transcription factor，CARF）降低其circRNAs的转录形式的水平（P<0.05）参与子宫内膜的发育；差异表达circRNAs的宿主基因GO和KEGG分析结果显示注释到紧密连接（Tight junctions）条目上和鸟苷三磷酸酶（GTPases）通路上的circRNAs参与了奶山羊子发情周期中非妊娠子宫内膜的发育过程；另外，本研究中得到的circRNAs与公共数据库中的436个山羊miRNAs有靶向结合位点。结论：奶山羊发情第5天和发情第15天的非妊娠子宫内膜组织中存在差异的circRNAs，生物信息学分析表明一些circRNAs参与奶山羊发情周期非妊娠子宫内膜的发育过程；本研究构建了奶山羊发情周期非妊娠子宫内膜circRNAs文库，丰富了奶山羊的转录组信息，有助于我们研究奶山羊子宫内膜发育过程中的分子调控机制，为进一步提高奶山羊的胚胎着床的成功率和繁殖率提供理论依据。创新点：本研究选择发情第5天和发情第15天奶山羊的非妊娠子宫内膜组织进行circRNAs表达谱分析；采用核糖体去除和链特异性文库构建方案进行RNA-seq测序，保留了完整的circRNAs序列，对样本中几乎全部的circRNAs序列进行鉴定和分析。

地球磁场影响着地球上所有的生物。本研究以正常地磁场（约50 μT）为对照条件，在近零磁场（<500 nT）下饲养粘虫，观察近零磁场对粘虫的行为效应。近零磁场由亥姆霍兹线圈系统产生。结果表明：近零磁场显著延长了粘虫幼虫和蛹的发育历期，增加了雄蛾的寿命，减少了蛹重，雌性产卵量和卵黄蛋白原蛋白基因（Vg）的表达水平；而且，近零磁场对粘虫成虫交配率产生负面影响。此外，近零磁场下粘虫在黑夜的自主飞行活动减少，表明近零磁场可能影响粘虫的飞行节律。磁场强度的降低可能对粘虫的生长发育、繁殖和飞行等活动产生负面影响，进而影响粘虫的迁飞活动。

Resistance gene analog (RGA) screening of mapped disease-resistant genes not only helps to clone these genes but also helps to develop efficient molecular markers for resistance breeding. The present study focused on the PmU region located on chromosome 7A^uL of Triticum urartu, and recently, a nucleotide binding site (NBS)-encoding gene, Pm60, was cloned from the same chromosome arm. In this research, NBS, protein kinase (PK), and ATP-binding cassette (ABC), the three disease resistance-related gene families, were analyzed within PmU region by using informatics tools, and an expression experiment was conducted to verify their functions in vivo. Comparative genomic analysis revealed that 126 RGAs were included on chromosome 7A^uL, and 30 of the RGAs as well as Pm60 were found in the PmU region. Transcriptome database analysis of T. urartu revealed 14 PmU-RGAs with expression data, and three PmU-NBSs exhibited significant changes in expression after inoculation with Blumeria graminis f. sp. tritici (Bgt); TRIUR3_14879 was up-regulated, while TRIUR3_00450 and TRIUR3_06270 were down-regulated. Cluster analysis showed that these three PmU-NBSs were clustered far from the cloned wheat resistance genes. Then, qRT-PCR was performed to investigate the expression of 14 PmU-RGAs and Pm60 after inoculation with Bgt race E09; the results showed that Pm60 was specifically expressed in UR206 which carrying PmU, but not in susceptible UR203; while TRIUR3_14879 was significantly up-regulated and TRIUR3_00450 was downregulated in UR206 after inoculation. These results indicated that PmU is Pm60, and TRIUR3_14879 and TRIUR3_00450 may also be involved in the defense against Bgt.

Cotton cultivars with brown (Xiangcaimian 2), green (Wanmian 39) and white (Sumian 9) fiber were investigated to study fiber developmental characteristics of natural-colored cotton and the effect of hormones on fiber quality at different stages after anthesis. Fiber lengths of both natural-colored cottons were lower than the white-fibered control, with brown-fibered cotton longer than green. Fiber strength, micronaire and maturation of natural-colored cotton were also lower than the control. The shorter fiber of the green cultivar was due to slower growth during 10 to 30 days post-anthesis (DPA). Likewise, the lower fiber strength, micronaire and maturation of natured-colored cotton were also due to slower growth during this pivotal stage. Indole-3-acetic acid (IAA) content at 10 DPA, and abscisic acid (ABA) content at 30 to 40 DPA were lower in the fibers of the natural-colored than that of the white-fibered cotton. After applying 20 mg L^–1 gibberellic acid (GA3), the IAA content at 20 DPA in the brown and green-fibered cottons increased by 51.07 and 64.33%, fiber ABA content increased by 38.96 and 24.40%, and fiber length increased by 8.13 and 13.96%, respectively. Fiber strength, micronaire and maturation were also enhanced at boll opening stage. Those results suggest that the level of endogenous hormones affect fiber quality. Application of external hormones can increase hormone content in natural-colored cotton fiber, improving its quality.

    Tassel branch number (TBN) is the principal component of maize tassel inflorescence architecture and is a typical quantitative trait controlled by multiple genes. The main objective of this research was to detect quantitative trait loci (QTLs) for TBN. The maize inbred line SICAU1212 was used as the common parent to develop BC₁S₁ and recombinant inbred line (RIL) populations with inbred lines 3237 and B73, respectively. The two related populations consisted of 123 and 238 lines, respectively. Each population was grown and phenotyped for TBN in two environments. Eleven QTLs were detected in the BC₁S₁ population, located on chromosomes 2, 3, 5, and 7, accounted for 4.45–26.58% of the phenotypic variation. Two QTLs (qB11Jtbn2-1, qB12Ctbn2-1, qBJtbn2-1; q11JBtbn5-1, qB12Ctbn5-1, qBJtbn5-1) that accounted for more than 10% of the phenotypic variation were identified. Three QTLs located on chromosomes 2, 3 and 5, exhibited stable expression in the two environments. Ten QTLs were detected in the RIL population, located on chromosomes 2, 3, 5, 8, and 10, accounted for 2.69–13.58% of the TBN variation. One QTL (qR14Dtbn2-2) explained >10% of the phenotypic variation. One common QTL (qB12Ctbn2-2, qR14Dtbn2-2, qRJtbn2-2) was detected between the two related populations. Three pairs of epistatic effects were identified between two loci with or without additive effects and accounted for 1.19–4.26% of the phenotypic variance. These results demonstrated that TBN variation was mainly caused by major effects, minor effects and slightly modified by epistatic effects. Thus, identification of QTL for TBN may help elucidate the genetic basis of TBN and also facilitate map-based cloning and marker-assisted selection (MAS) in maize breeding programs.

    The pleiotropic drug resistance (PDR) sub-family of adenosine triphosphate (ATP)-binding cassette (ABC) transporter had been reported to participate in diverse biological processes of plant. In this study, we cloned three novel PDR genes in Fusarium head blight (FHB) resistant wheat cultivar Ning 7840, which were located on wheat chromosomes 6A, 6B and 6D. In phylogeny, these genes were members of cluster I together with AePDR7 and BdPDR7. Subcellular localization analysis showed that TaPDR7 was expressed on the plasmalemma. The quantitative real time PCR (RT-PCR) analysis showed that this gene and its probable orthologues in chromosomes 6B and 6D were both up-regulated sharply at 48 h after infected by Fusarium graminearum and trichothecene deoxynivalenol (DON) in spike. When knocking down the transcripts of all TaPDR7 members by barely stripe mosaic virus-induced gene silencing (BSMV-VIGS) system, it could promote the F. graminearum hyphae growth and made larger pathogen inoculation points in Ning 7840, which suggested that TaPDR7 might play an important role in response to F. graminearum. Although salicylic acid (SA), methyl jasmonate (MeJA) and abscisic acid (ABA) had been reported to possibly regulate wheat FHB resistance, here, we found that the three members of TaPDR7 were negatively regulated by these three hormones but positively regulated by indoleacetic acid (IAA).

Spermatogonial stem cells (SSCs) form the foundation for spermatogenesis and sustain male fertility. To explore the regulatory mechanisms of chicken SSCs generation, we obtained highly purified chicken embryonic stem cells (ESCs), primordial germ cells (PGCs) and SSCs by fluorescence-activated cell sorting (FACS). High-throughput analysis methods (RNA-Seq) were used to sequence the transcriptome level of these cells. Gene ontology and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment were used to analyze RNA-Seq results. BMP4 was used to induce chicken ESCs differentiation to SSCs-like cells in vitro. The quantitative real-time (qRT)-PCR was used to detect the expression changes of the key genes. The results showed that 22 relevant critical pathways were found by RNA-Seq, one of them was the Janus kinase/signal transducer and activator of transcription (JAK/STAT) signaling pathway. Total of 103 related genes were detected in this pathway. Protein-protein interactions analysis found that 87 proteins were significantly related to 19 key proteins in this pathway. These 87 proteins were enriched in 21 biological processes and 18 signaling pathways. Moreover, during the differentiation of chicken ESCs to SSCs-like cells induced by BMP4 in vitro, JAK2 and STAT3 were activated. The qRT-PCR results showed that the expression trends of JAK2 and STAT3 were basically the same as in vivo. We concluded that JAK/STAT signaling pathway plays an important role in the process of chicken SSCs generation both in vivo and in vitro; it may achieve its function through multiple biological processes and other related pathways.

A endochitinase gene (Tch) from the fungus Trichoderma viride was introduced into broccoli (Brassica oleracea var. italica) by Agrobacterium-mediated transformation. Sixty-eight putative transformants were obtained and the presence of the Tch gene was confirmed by both PCR and Southern blot analysis. RT-PCR analysis showed an accumulation of the transcript encoding the endochitinase protein in the transgenic plants. Using real-time quantitative PCR, the expression profiling of endochitinase gene was analyzed. Primary transformants and selfed progeny were examined for expression of the endochitinase using a fluorometric assay and for their resistance to the pathogenic fungi Botrytis cinerea and Rhizoctonia solani. The endochitinase activities in T0 in vitro plants, T0 mature plants and T1 mature plants were correlated with leaf lesions, and the transgenic line T618 had high endochitinse activities of 102.68, 114.53 and 120.27 nmol L–1 MU min–1 mg–1 protein in the three kinds of plants, respectively. The endochitinase activity showed a positive correlation with the resistance to the pathogens. Most transgenic T0 broccoli had increased resistance to the pathogens of B. cinerea and R. solani in leaf assays and this resistance was confirmed to be inheritable. These findings suggested that expression of the Tch gene from T. viride could enhance resistance to pathogenic fungi in Brassica species.

A novel phytoplasma was detected in a cherry plum (Prunus cerasifera Ehrh) tree that mainly showed yellow leaf symptom. The tree was growing in an orchard located in Yangling District, Shaanxi Province, China. The leaves started as chlorotic and yellowing along leaf minor veins and leaf tips. Chlorosis rapidly developed to inter-veinal areas with the whole leaf becoming pale yellow in about 1-4 wk. Large numbers of phytoplasma-like bodies (PLBs) were seen under transmission electron microscopy. The majority of the PLBs was spherical or elliptical vesicles, with diameters in range of 0.1-0.6 μm, and distributed in the phloem cells of the infected tissues. A 1246-bp 16S ribosomal RNA (rRNA) gene fragment was amplified from DNA samples extracted from the yellow leaf tissues using two phytoplasma universal primer pairs R16mF2/R16mR1 and R16F2n/R16R2. Phylogenetic analysis using the 16S rRNA gene sequence suggested that the phytoplasma associated with the yellow leaf symptoms belongs to a novel subclade in the aster yellows (AY) group (16SrI group). Virtual and actual restriction fragment length polymorphism (RFLP) analysis of the 16S rRNA gene fragment revealed that the phytoplasma was distinguishable from all existing 19 subgroups in the AY group (16SrI) by four restriction sites, Hinf I, Mse I, Sau3A I and Taq I. The similarity coefficients of comparing the RFLP pattern of the 16S rRNA gene fragment of this phytoplasma to each of the 19 reported subgroups ranged from 0.73 to 0.87, which indicates the phytoplasma associated with the cherry plum yellow leaf (CPYL) symptoms is probably a distinct and novel subgroup lineage in the AY group (16SrI). In addition, the novel phytoplasma was experimentally transmitted to periwinkle (Catharanthus roseus) plants from the tree with CPYL symptoms and then back to a healthy 1-yr-old cherry plum tree via dodder (Cuscuta odorata) connections.