鼠兔核质杂交胚胎早期发育的研究

细胞核移植技术已被证明是研究发育中核质相互关系的非常重要的手段之一,电融合技术也是近十年发展起来的新型细胞融合技术。本实验运用这两项技术,进行了鼠、兔目间核质杂交实验,小鼠8-细胞核在激活的兔去核卵母细胞中,发生了染色体超前凝聚及核膨胀,融合卵移植到小鼠输卵管4.5天后,冲洗出,有5.4%的重构卵发育到囊胚期,通过染色体检查,囊胚细胞中均为小鼠染色体,其中一个囊胚为正常小鼠核型(2 n=40,XX)。通过本实验,我们认为:鼠兔远缘核质杂交胚胎的早期发育是可能的。     

TTLL11 gene is associated with sustained attention performance and brain networks: A genome-wide association study of a healthy Chinese sample

Genetic studies on attention have mainly focused on children with attention-deficit/hyperactivity disorder (ADHD), so little systematic research has been conducted on genetic correlates of attention performance and their potential brain mechanisms among healthy individuals. The current study included a genome-wide association study (GWAS, N = 1145 healthy young adults) aimed to identify genes associated with sustained attention and an imaging genetics study (an independent sample of 483 healthy young adults) to examine any identified genes' influences on brain function. The GWAS found that TTLL11 showed genome-wide significant associations with sustained attention, with rs13298112 as the most significant SNP and the GG homozygotes showing more impulsive but also more focused responses than the A allele carriers. A retrospective examination of previously published ADHD GWAS results confirmed an un-reported, small but statistically significant effect of TTLL11 on ADHD. The imaging genetics study replicated this association and showed that the TTLL11 gene was associated with resting state activity and connectivity of the somatomoter network, and can be predicted by dorsal attention network connectivity. Specifically, the GG homozygotes showed lower brain activity, weaker brain network connectivity, and non-significant brain-attention association compared to the A allele carriers. Expression database showed that expression of this gene is enriched in the brain and that the G allele is associated with lower expression level than the A allele. These results suggest that TTLL11 may play a major role in healthy individuals' attention performance and may also contribute to the etiology of ADHD.     

Transmembrane kinase 1-mediated auxin signal regulates membrane-associated clathrin in Arabidopsis roots

Clathrin-mediated endocytosis (CME) is the major endocytic pathway in eukaryotic cells that directly regulates abundance of plasma membrane proteins. Clathrin triskelia are composed of clathrin heavy chains (CHCs) and light chains (CLCs), and the phytohormone auxin differentially regulates membrane-associated CLCs and CHCs, modulating the endocytosis and therefore the distribution of auxin efflux transporter PIN-FORMED2 (PIN2). However, the molecular mechanisms by which auxin regulates clathrin are still poorly understood. Transmembrane kinase (TMKs) family proteins are considered to contribute to auxin signaling and plant development; it remains unclear whether they are involved in PIN transport by CME. We assessed TMKs involvement in the regulation of clathrin by auxin, using genetic, pharmacological, and cytological approaches including live-cell imaging and immunofluorescence. In tmk1 mutant seedlings, auxin failed to rapidly regulate abundance of both CHC and CLC and to inhibit PIN2 endocytosis, leading to an impaired asymmetric distribution of PIN2 and therefore auxin. Furthermore, TMK3 and TMK4 were shown not to be involved in regulation of clathrin by auxin. In summary, TMK1 is essential for auxin-regulated clathrin recruitment and CME. TMK1 therefore plays a critical role in the establishment of an asymmetric distribution of PIN2 and an auxin gradient during root gravitropism.     

Universal two-dimensional labelled probe-mediated melting curve analysis based on multiplex PCR for rapid typing of Plasmodium in a single closed tube

Currently, malaria is still one of the major public health problems commonly caused by the four Plasmodium species. The similar symptoms of malaria and the COVID-19 epidemic of fever or fatigue lead to frequent misdiagnosis. The disadvantages of existing detection methods, such as time-consuming, costly, complicated operation, need for experienced technicians, and indistinguishable typing, lead to difficulties in meeting the clinical requirements of rapid, easy, and accurate typing of common Plasmodium species. In this study, we developed and optimized a universal two-dimensional labelled probe-mediated melting curve analysis (UP-MCA) assay based on multiplex and asymmetric PCR for rapid and accurate typing of five Plasmodium species, including novel human Plasmodium, Plasmodium knowlesi (Pk), in a single closed tube following genome extraction. The assay showed a limit of detection (LOD) of 10 copies per reaction and could accurately distinguish Plasmodium species from intra-plasmodium and other pathogens. Additionally, we proposed and validated different methods of fluorescence quenching and tag design for probes that are suitable for UP-MCA assays. Moreover, the clinical performance of the Plasmodium UP-MCA assay using a base-quenched universal probe was evaluated using 226 samples and showed a sensitivity of 100% (164/164) and specificity of 100% (62/62) at a 99% confidence interval, with the microscopy method as the gold standard. In summary, the UP-MCA assay showed excellent sensitivity, specificity, and accuracy for genotyping Plasmodium species spp. Additionally, it facilitates convenient and rapid Plasmodium detection in routine clinical practice and has great potential for clinical translation.     

Factors Affecting Plant Regeneration from Tissue Cultures of Chinese Leymus (Leymus chinensis)

Chinese leymus (Leymus chinensis Trin.) is a perennial grass of the Gramineae, which is widely distributed in China, Mongolia and in Russian-Siberian. In order to explore the potential of biotechnology for genetic improvement of this forage grass, an efficient tissue culture system was established and the factors affecting plant regeneration were evaluated. Immature inflorescence segments 3–5 mm in length from eight accessions were cultured on N6 medium supplemented with 2.26–22.60 µM 2,4-D. The callus induction frequency ranged from 72.11 to 82.19%. Shoots were differentiated from the calli on N6 medium containing 4.65 µM kinetin and 4.44 µM BA. Viable regenerants were developed on hormone-free medium. Normal plants were obtained after natural vernalization in the field. The plant regeneration frequency in Chinese leymus was associated with different genotypes and different combinations of growth regulators in medium. The concentration of 2,4-D in the callus induction medium had a strong effect on successive plant regeneration. Relatively higher concentrations of 2,4-D (i.e., 9.04 and 22.60 µM) were more favorable to the plant regeneration than lower ones (i.e., 2.26 and 4.52 µM). This is the first report on plant regeneration in vitro in L. chinensis.     

抗旱性不同的两个大豆品种对外源H_2O_2的响应

两个品种的大豆叶圆片经１０－４ｍｏｌ／Ｌ和１０－３ｍｏｌ／Ｌ的Ｈ２Ｏ２处理１２ｈ后,超氧物歧化酶（ＳＯＤ）、过氧化氢酶（ＣＡＴ）与谷胱甘肽还原酶（ＧＲ）活性明显增加,但１０－２ｍｏｌ／Ｌ的Ｈ２Ｏ２处理却使这些酶活性降低。抗旱性较强的大豆品种小粒豆１号较抗旱性较弱的鲁豆４号能维持较高的叶绿素含量和较高的ＳＯＤ、ＣＡＴ及ＧＲ活性,对Ｈ２Ｏ２的抗性较强。５０μｍｏｌ／Ｌ的亚胺环已酮（ＣＨＭ）能消除Ｈ２Ｏ２对ＳＯＤ、ＣＡＴ与ＧＲ活性的刺激作用,而同样浓度的放线菌素Ｄ（ＡＭＤ）则不能。