诱发小麦成熟胚愈伤组织及其再生植株抗盐性变异的研究

本研究以普通小麦成熟胚为起始材料,以平阳霉素(PYM)和正定霉素(ZDM)为诱变剂。发现不同基因型对药物的敏感性不同,其中对药物敏感的在含盐筛选培养基上的存活率高。在诱导培养基内添加一定量的诱变剂可以产生耐盐变异。这种抗性愈伤组织转入与筛选培养基含盐量相同的分化培养基,比较容易产生耐盐再生植株。其M_1、M_2代较亲本系表现株高降低,穗长变短、籽粒饱满度也差;M_1代的结实率仅有5.5%,M_2代恢复到40.9%。利用M_1代和亲本系实生苗叶片,进行脯氨酸含量测定。发现耐盐变异株在无盐的Hoagland溶液中,游离脯氨酸的含量超过亲本系,对盐胁迫不敏感,其耐受力强于对照,具有一定的抗盐稳定性。     

ploidyfrost: Reference-free estimation of ploidy level from whole genome sequencing data based on de Bruijn graphs

Polyploidy is ubiquitous and its consequences are complex and variable. A change of ploidy level generally influences genetic diversity and results in morphological, physiological and ecological differences between cells or organisms with different ploidy levels. To avoid cumbersome experiments and take advantage of the less biased information provided by the vast amounts of genome sequencing data, computational tools for ploidy estimation are urgently needed. Until now, although a few such tools have been developed, many aspects of this estimation, such as the requirement of a reference genome, the lack of informative results and objective inferences, and the influence of false positives from errors and repeats, need further improvement. We have developed ploidyfrost , a de Bruijn graph-based method, to estimate ploidy levels from whole genome sequencing data sets without a reference genome. ploidyfrost provides a visual representation of allele frequency distribution generated using the ggplot2 package as well as quantitative results using the Gaussian mixture model. In addition, it takes advantage of colouring information encoded in coloured de Bruijn graphs to analyse multiple samples simultaneously and to flexibly filter putative false positives. We evaluated the performance of ploidyfrost by analysing highly heterozygous or repetitive samples of Cyclocarya paliurus and a complex allooctoploid sample of Fragaria × ananassa. Moreover, we demonstrated that the accuracy of analysis results can be improved by constraining a threshold such as Cramér's V coefficient on variant features, which may significantly reduce the side effects of sequencing errors and annoying repeats on the graphical structure constructed.     

Peptide MSI-1 inhibited MCR-1 and regulated outer membrane vesicles to combat immune evasion of Escherichia coli

Polymyxin resistance is conferred by MCR-1 (mobile colistin resistance 1)-induced lipopolysaccharide (LPS) modification of G⁻ bacteria. However, the peptide MSI-1 exerts potent antimicrobial activity against mcr-1-carrying bacteria. To further investigate the potential role of MCR-1 in improving bacterial virulence and facilitating immune evasion, and the immunomodulatory effect of peptide MSI-1, we first explored outer membrane vesicle (OMV) alterations of mcr-1-carrying bacteria in the presence and absence of sub-MIC MSI-1, and host immune activation during bacterial infection and OMV stimulation. Our results demonstrated that LPS remodelling induced by MCR-1 negatively affected OMV formation and protein cargo by E. coli. In addition, MCR-1 diminished LPS-stimulated pyroptosis but facilitated mitochondrial dysfunction, further aggravating apoptosis in macrophages induced by OMVs of E. coli. Similarly, TLR4-mediated NF-κB activation was markedly alleviated once LPS was modified by MCR-1. However, peptide MSI-1 at the sub-MIC level inhibited the expression of MCR-1, further partly rescuing OMV alteration and attenuation of immune responses in the presence of MCR-1 during both infection and OMV stimulation, which can be exploited for anti-infective therapy.     

High-resolution mapping of the S and Z loci of Phalaris coerulescens.

Self incompatibility (SI) in Phalaris coerulescens is gametophytically determined by two unlinked multi allelic loci (S and Z). Neither the S nor Z genes have yet been cloned. As part of a map-based cloning strategy, high-resolution maps of the S and Z regions were generated from distorted segregating populations using RFLP probes from wheat, barley, oat, and Phalaris. The S locus was delimited to 0.26 cM with two boundary markers (Xwg811 and Xpsr168) and cosegregated with Xbm2 and Xbcd762. Xbcd266 was the closest marker linked to Z (0.9 cM). A high level of colinearity in the S and Z regions was found in both self-incompatible and -compatible species. The S locus was localized to the subcentromere region of chromosome 1 and the Z locus to the long arm end of chromosome 2. Several rice BAC clones orthologous to the S and Z locus regions were identified. This opens the possibility of using the rice genome sequence data to generate more closely linked markers and identify SI candidate genes. These results add further support to the conservation of gene order in the S and Z regions of the grass genomes.     

高原适应过程中大鼠脑内β－内啡肽含量变化的研究

选用成年雄性Ｗｉｓｔａｒ大鼠,由平原（海拔５ｍ,上海）直接运送到高原（２２６１ｍ,西宁和３４６０ｍ,天峻）。以放射免疫分折法研究了大鼠在高原２４ｈ．急性缺氧期和３０ｄ慢性缺氧期中枢脑内β－内啡肽样免疫活性物质（β－ＥＰ）的含量变化。结果表明：１．大鼠在两个不同海拔（２２６１ｍ,３４６０ｍ）环境２４ｈ急性缺氧期与平原对照组相比,脑垂体内β－ＥＰ含量降低非常显著（Ｐ＜０．０１）,纹状体、丘脑、下丘脑、桥－延脑、海马内β－ＥＰ含量均增加非常显著（Ｐ＜０．０１）。２．大鼠在３４６０ｍ高原３０ｄ慢性习服期,垂体及各脑区内β－ＥＰ的含量变化呈时相性：即１—３ｄ均呈进行性增加（Ｐ＜０．０１）：３－１５ｄ呈持续性减少,除中脑、纹状体、丘脑外,均为Ｐ＜０．０１。１５－３０ｄ垂体、丘脑、皮层、纹状体内β－ＥＰ含量仍持续减少（毒体、皮层Ｐ＜０．０１）,桥－延脑、下丘脑、海马趋于回升（Ｐ＜０．０１）,中脑亦趋于回升（Ｐ＞０．０５）。脑内β－ＥＰ的这种变化可能具有十分重要的生物学意义。     

用地高辛标记探针检测基因工程人β干扰素制品中的外源DNA

基因工程人β干扰素工程菌经发酵培养、纯化后获得纯的制品,采用斑点杂交技术,用非放射性地高辛标记超声裂解的全工程菌ＤＮＡ作为探针,检测基因工程人β干扰素纯品,结果显示：基因工程人β干扰素纯品中的外源ＤＮＡ含量小于１００ｐｇ／剂。     

鼠肝DNA甲基化酶的纯化及物理化学性质的研究

以Ｗｉｓｔａｒ大鼠肝为材料,确立了一个简便的纯化鼠肝ＤＮＡ甲基化酶的程序,包括：细胞的超声破碎、去内源核酸、硫酸铵盐析、磷酸纤维素亲和层析、ＤＥＡＥ－ＳｅｐｈａｄｅｘＡ－５０柱层析及ＳｅｐｈａｄｅｘＧ－１５０凝胶过滤。用不同浓度聚丙烯酰胺凝胶电泳和孔梯度凝胶电泳检测,纯化后的酶已达电泳均一,且酶的比活力提高１１２倍。以聚丙烯酰胺孔梯度凝胶电泳测得其天然酶的分子量为３６５ｋＤ,以ＳＤＳ－聚丙烯酰胺凝胶电泳测得该酶有两种亚基,大亚基为９５ｋＤ,小亚基为８５ｋＤ,推测该酶由两个大亚基和两个小亚基组成。     

Aliphatic nitro-compounds in Astragalus canadensis

A reinvestigation of the alphatic nitro-compounds in Astragalus canadensis resulted in the identification of two new esters of glucose with 3-nitropropanoic acid and 5-oxotetrahydrofuran-3-acetic acid, together with six known conjugates of 3-nitropropanoic acid. ¹H and ¹³CNMR data are reported for the new compounds.