间斑寇蛛卵粒毒素-Ⅵ的基因克隆、异源表达与活性鉴定

间斑寇蛛Latrodectus tredecimguttatus的一个显著特点是其毒腺外组织甚至卵粒中也存在毒性成分。研究毒腺外的毒素不但可以加深对蜘蛛毒素的了解,而且可以发现具有重要应用前景的新型毒素分子。为了探究间斑寇蛛卵粒中低丰度表达的蛋白质类毒素,利用生物信息学方法从间斑寇蛛卵粒转录组中挖掘出一条编码多肽毒素的基因序列,利用基于3′-RACE和巢式PCR的策略成功克隆并异源表达了该基因。表达的多肽毒素命名为间斑寇蛛卵粒毒素-Ⅵ(Latroeggtoxin-Ⅵ,LETX-Ⅵ)。生物学活性鉴定结果表明,LETX-Ⅵ能抑制ND7/23细胞膜上的钠离子通道电流和促进PC12细胞多巴胺的释放,但对美洲蜚蠊Periplaneta americana和金黄色葡萄球菌及白色念珠菌等细菌和真菌不显示明显的毒性,说明LETX-Ⅵ是一种哺乳动物特异的神经毒素,在神经生物学研究工具试剂和相关疾病治疗药物的研发等方面具有潜在的应用前景。     

Molecular mapping of leaf rust resistance gene LrNJ97 in Chinese wheat line Neijiang 977671

Neijiang 977671 and 19 near-isogenic lines with known leaf rust resistance genes were inoculated with 12 pathotypes of Puccinia triticina for postulation of leaf rust resistance genes effective at the seedling stage. The reaction pattern of Neijiang 977671 differed from those of the lines with known leaf rust resistance genes used in the test, indicating that Neijiang 977671 may carry a new leaf rust resistance gene(s). With the objective of identifying and mapping the new gene for resistance to leaf rust, F₁ and F₂ plants, and F_2:3 families, from Neijiang 977671 × Zhengzhou 5389 (susceptible) were inoculated with Chinese P. triticina pathotype FHNQ in the greenhouse. Results from the F₂ and F_2:3 populations indicated that a single dominant gene, temporarily designated LrNJ97, conferred resistance. In order to identify other possible genes in Neijiang 977671 other eight P. triticina pathotypes avirulent on Neijiang 977671 were used to inoculate 25 F_2:3 families. The results showed that at least three leaf rust resistance genes were deduced in Neijiang 977671. Bulked segregant analysis was performed on equal amounts of genomic DNA from 20 resistant and 20 susceptible F₂ plants. SSR markers polymorphic between the resistant and susceptible bulks were used to analyze the F_2:3 families. LrNJ97 was linked to five SSR loci on chromosome 2BL. The two closest flanking SSR loci were Xwmc317 and Xbarc159 at genetic distances of 4.2 and 2.2 cM, respectively. At present two designated genes (Lr50 and Lr58) are located on chromosome 2BL. In the seedling tests, the reaction pattern of LrNJ97 was different from that of Lr50. Lr50 and Lr58 were derived from T. armeniacum and Ae. triuncialis, respectively, whereas according to the pedigree of Neijiang 977671 LrNJ97 is from common wheat. Although seeds of lines with Lr58 were not available, it was concluded that LrNJ97 is likely to be a new leaf rust resistance gene.     

Gene cloning and functional analysis of yellow green leaf3 (ygl3) gene during the whole-plant growth stage in rice

Chlorophyll is an important photosynthetic pigment in the process of photosynthesis in plants and photosynthetic bacteria. Genes involved in chlorophyll biosynthesis in Arabidopsis and photosynthetic bacteria have been well documented. In rice, however, these genes have not been fully annotated. In this paper, a yellow-green leaf gene, yellow green leaf3 (ygl3) was cloned and analyzed. ygl3 encodes magnesium chelation ChlD (D) subunit, a key enzyme for chlorophyll synthesis, resulting in a yellow-green leaf phenotype in all growth stages in rice. Expression content of ygl3 is highest in the leaf blades, followed by the leaf sheaths, while there is virtually no expression of the gene in the stems and seeds. The sub-cellular structure and protein content of the photosynthetic system of the ygl3 mutant were revealed by transmission electron microscopy, BN-PAGE, and western blotting. The results show that the mutation of the ygl3 gene indirectly leads to a decrease in the protein content of the photosynthetic system and severely obstructs the formation of granum thylakoids.     

中国水韭属两个四倍体新种

水韭属(Isoëtes)是起源最为古老的水生维管植物,全属物种均被列为国家一级重点保护植物。通过对全国水韭属植物的调查和研究发现,不同产地的四倍体植株在形态上存在显著差异。基于形态学、孢粉学和细胞学证据,将分布于中国湖南省长沙地区和怀化地区的四倍体居群分别命名为隆平水韭(Isoëtes longpingii)和湘妃水韭(I. xiangfei),并详细描述了其形态特征。隆平水韭形态上与中华水韭(I. sinensis)相似,不同之处在于其大孢子具小的瘤状或冠状纹饰,叶细长而柔弱,长达60 cm; 该种也与六倍体东方水韭(I. orientalis)相似,不同之处在于其染色体44条,大孢子具瘤状或冠状纹饰。湘妃水韭的大孢子纹饰虽与二倍体云贵水韭(I. yunguiensis)相似,但在小孢子纹饰、孢子囊形状和染色体数目方面却不同。隆平水韭仅少数植株生长于湖南省宁乡市一处池塘,完全沉水生长,而湘妃水韭则分布于怀化市通道县和会同县的湿地。由于这两个新种的分布区狭窄,野生居群数量和个体数较少,栖息地环境受到人为干扰,因此根据IUCN红色名录评估标准,将隆平水韭评为极危(CR)等级,湘妃水韭评为易危(VU)等级。所编制的中国已知水韭属物种的分种检索表,为本属物种的鉴定和保护工作提供了重要参考。     

Adenohypophysis formation in the zebrafish and its dependence on sonic hedgehog

Formation of the adenohypophysis in mammalian embryos occurs via an invagination of the oral ectoderm to form Rathke's pouch, which becomes exposed to opposing dorsoventral gradients of signaling proteins governing specification of the different hormone-producing pituitary cell types. One signal promoting pituitary cell proliferation and differentiation to ventral cell types is Sonic hedgehog (Shh) from the oral ectoderm. To study pituitary formation and patterning in zebrafish, we cloned four cDNAs encoding different pituitary hormones, prolactin (prl), proopiomelancortin (pomc), thyroid stimulating hormone (tsh), and growth hormone (gh), and analyzed their expression patterns relative to that of the pituitary marker lim3. prl and pomc start to be expressed at the lateral edges of the lim3 expression domain, before pituitary cells move into the head. This indicates that patterning of the pituitary anlage and terminal differentiation of pituitary cells starts while cells are still organized in a placodal fashion at the anterior edge of the developing brain. Following the expression pattern of prl and pomc during development, we show that no pituitary-specific invagination equivalent to Rathke's pouch formation takes place. Rather, pituitary cells move inwards together with stomodeal cells during oral cavity formation, with medial cells of the placode ending up posterior and lateral cells ending up anterior, resulting in an anterior-posterior, rather than a dorsoventral, patterning of the adenohypophysis. Carrying out loss- and gain-of-function experiments, we show that Shh from the ventral diencephalon plays a crucial role during induction, patterning, and growth of the zebrafish adenohypophysis. The phenotypes are very similar to those obtained upon pituitary-specific inactivation or overexpression of Shh in mouse embryo, suggesting that the role of Shh during pituitary development has been largely conserved between fish and mice, despite the different modes of pituitary formation in the two vertebrate classes.     

骆驼瘤胃耐受/降解去氢骆驼蓬碱细菌的筛选

【目的】中药骆驼蓬含多种生物碱,对动物有毒性。生活于荒漠半荒漠的骆驼可采食部分有毒植物而不中毒。为了解骆驼瘤胃微生物对骆驼蓬植物毒素的耐受与降解能力进行本研究。【方法】以含100mg/L纯品去氢骆驼蓬碱的M98-5培养基接种骆驼瘤胃内容物,经五代胁迫培养后分离可耐受/降解去氢骆驼蓬碱的细菌,以薄层析法检验其降解活力,以16SrRNA序列分析其进化地位。【结果】29个分离株中15株具有降解去氢骆驼蓬碱活性;16SrRNA序列分析显示,属于乳杆菌属(Lactobacillus)16株,占55%;志贺氏菌属(Shigella)7株,占24%;芽孢杆菌属(Bacillus)4株,占13.8%;肠球菌属(Enterococcus)和巨型肠球菌属(Megasphaera)各1株。【结论】可耐受/降解去氢骆驼蓬碱的骆驼瘤胃细菌仅限于少数几类,且检测到的具有降解活力的只有乳杆菌类。