A leaf shape mutant provides insight into PINOID Serine/Threonine Kinase function in cucumber(Cucumis sativus L.)

Optimizing leaf shape is a major challenge in efforts to develop an ideal plant type. Cucumber leaf shapes are diverse; however, the molecular regulatory mechanisms underlying leaf shape formation are unknown. In this study, we obtained a round leaf mutant(rl) from an ethyl methanesulfonate-induced mutagenesis population. Genetic analysis revealed that a single recessive gene, rl, is responsible for this mutation. A modified Mut Map analysis combined linkage mapping identified a single nucleotide polymorphism within a candidate gene,Csa1 M537400, as the mutation underlying the trait.Csa1 M537400 encodes a PINOID kinase protein involved in auxin transport. Expression of Csa1 M537400 was significantly lower in the rl mutant than in wild type, and it displayed higher levels of IAA(indole-3-acetic acid) in several tissues. Treatment of wild-type plants with an auxin transport inhibitor induced the formation of round leaves,similar to those in the rl mutant. Altered expression patterns of several auxin-related genes in the rl mutant suggest that rl plays a key role in auxin biosynthesis,transport, and response in cucumber. These findings provide insight into the molecular mechanism underlying the regulation of auxin signaling pathways in cucumber,and will be valuable in the development of an ideal plant type.     

Plasticity of mycangia in Xylosandrus ambrosia beetles

Insects that depend on microbial mutualists evolved a variety of organs to transport the microsymbionts while dispersing. The ontogeny and variability of such organs is rarely studied, and the microsymbiont*s effects on the animal tissue development remain unknown in most cases. Ambrosia beetles (Coleoptera: Curculionidae: Scolytinae or Platypodinae) and their mutualistic fungi are an ideal system to study the animalfungus interactions. While the interspecific diversity of their fungus transport organ一 mycangia—is well-known, their developmental plasticity has been poorly described. To determine the ontogeny of the mycangium and the influence of the symbiotic fungus on the tissue development, we dissected by hand or scanned with micro-CT the mycangia in various developmental stages in five Xylosandrus ambrosia beetle species that possess a large, mesonotal mycangium: Xylosandrus amputatus. Xylosandrus compactus, Xylosandrus crassiusculus, Xylosandrus discolor, and Xylosandrus germanus. We processed 181 beetle samples from the United States and China. All five species displayed three stages of the mycangium development:(1) young teneral adults had an empty, deflated and cryptic mycangium without fungal mass;(2) in fully mature adults during dispersal, the promesonotal membrane was inflated, and most individuals developed a mycangium mostly filled with the symbiont, though size and symmetry varied;and (3) after successful establishment of their new galleries, most females discharged the bulk of the fun gal inoculum and deflated the mycangium. Experimental aposymbiotic individuals demonstrated that the pronotal membrane invaginated independently of the presence of the fungus, but the fungus was required for inflation. Mycangia are more dynamic than previously thought, and their morphological changes correspond to the phases of the symbiosis. Importantly, studies of the fungal symbionts or plant pathogen transmission in ambrosia beetles need to consider which developmental stage to sample. We provide illustrations of the different stages, including microphotography of dissections and micro-CT scans.     

Genome editing in Bombyx mori: New opportunities for silkworm functional genomics and the sericulture industry

In recent years, research in life sciences has been remarkably revolutionized owing to the establishment, development and application of genome editing technologies. Genome editing has not only accelerated fundamental research but has also shown promising applications in agricultural breeding and therapy. In particular, the clustered, regularly interspaced, short palindromic repeat (CRISPR) technology has become an indispensable tool in molecular biology owing to its high efficacy and simplicity. Genome editing tools have also been established in silkworm (Bombyx mori), a model organism of Lepidoptera insects with high economic importance. This has remarkably improved the level and scope of silkworm research and could reveal new mechanisms or targets in basic entomology and pest management studies. In this review, we summarize the progress and potential of genome editing in silkworm and its applications in functional genomic studies for generating novel genetic materials.     

动力髋螺钉与股骨近端髓内钉内固定治疗股骨粗隆间骨折患者的疗效及安全性和关节功能对比观察

目的:比较动力髋螺钉(DHS)与股骨近端髓内钉(PFNA)内固定治疗股骨粗隆间骨折患者的疗效及安全性和关节功能。方法:选取滁州市第一人民医院于2013年3月～2018年4月期间收治的160例股骨粗隆间骨折患者,根据内固定方式的不同将患者分为DHS组(n=80,采用DHS内固定)和PFNA组(n=80,采用PFNA内固定),比较两组临床疗效,采用髋关节功能Harris评分评价所有患者关节功能恢复情况,比较两组患者术前及术后相关指标,并观察患者术后并发症发生情况。结果:PFNA组患者临床总有效率为90.00%,高于DHS组患者的68.75%(P0.05)。两组患者Harris评分的优良率比较差异无统计学意义(P0.05)。PFNA组患者手术时间、卧床时间、骨折愈合时间、切口长度均短于DHS组(P0.05),术中出血量、术后引流量均少于DHS组(P0.05)。两组患者术后并发症总发生率比较无统计学差异(P0.05)。结论:DHS与PFNA内固定治疗股骨粗隆间骨折在术后关节功能恢复、安全性方面效果相当,但与DHS内固定治疗比较,PFNA内固定治疗的临床疗效更佳,手术时间更短,出血量更少,患者术后恢复更快,是治疗股骨粗隆间骨折较理想的手术方式。     

水稻减数分裂染色体分析方法

减数分裂粗线期染色体研究技术的发展, 很大程度上克服了水稻(Oryza sativa)细胞遗传研究中较小染色体所带来的研究困难。减数分裂染色体的制备与观察已经成为水稻细胞遗传学研究中的常规方法。该文详细描述了水稻中常用的减数分裂染色体制备、荧光原位杂交和免疫荧光染色的实验方法。     

INH-α对BMP9诱导间充质干细胞成骨分化的作用

为了研究抑制素α亚基(inhibinα-subunit INH-α)对骨形态发生蛋白9(bone morphogenetic protein9,BMP9)诱导的间充质干细胞(mesenchymal stem cells,MSCs)成骨分化的影响,本研究采用细胞化学染色法检测第3天、第5天、第7天细胞中碱性磷酸酶(alkaline phosphatase,ALP)活性的变化。利用RT-PCR和Western blotting检测细胞中的成骨分化早期标志物(Runx2)和晚期标志物(OPN)的mRNA含量及蛋白表达水平。茜素红S染色法检测第21天细胞中的钙盐沉积变化。发现BMP9组ALP活性明显增高,INH-α组ALP活性与对照组相比无明显变化,但联合运用BMP9和INH-α组ALP活性较BMP9组明显降低。此外,BMP9组Runx2和OPN的mRNA含量和蛋白表达水平明显增高,而联用BMP9和INH-α组中的Runx2和OPN水平较BMP9组显著下降(p<0.01)。同样,在茜素红S染色实验中,BMP9组钙盐结节明显增多,染色深;而在联合运用BMP9和INH-α组钙盐结节较BMP9组明显减少,染色变浅。说明INH-α能够抑制BMP9诱导间充质干细胞成骨分化作用。     

Integrated analysis of gene expression and methylation profiles of novel pancreatic cancer cell lines with highly metastatic activity

Pancreatic cancer is one of the most lethal human malignancies, partly because of its propensity for metastasis. However, highly metastatic human pancreatic cancer cell lines suitable for studies of metastasis are currently lacking. Here we established two highly metastatic human pancreatic cancer cell lines, MIA PaCa-2 In8 and Panc-1 In8, by Matrigel induction assay. The cell lines were further characterized both in vitro and in vivo. MIA PaCa-2 In8 and Panc-1 In8 cells demonstrated increased migration and invasion compared with their respective parental cells. Following injection into nude mice, MIA PaCa-2 In8 and Panc-1 In8 cells resulted in more pulmonary metastases compared with the parental cells. Furthermore, analyses of m RNA, long non-coding RNA, micro RNA, and methylation profiling revealed that these factors were aberrantly regulated in the highly metastatic cells,indicating that they probably affected metastasis. We thus established and characterized two highly metastatic human pancreatic cell lines that could be used as valuable tools for future investigations into the pathogenesis, metastasis, and potential treatment of human pancreatic cancer.