Genome-wide analysis of ACO and ACS genes in pear (Pyrus ussuriensis)

The “Nanguo” pear is a typically climacteric fruit and ethylene is the main factor controlling the ripening process of climacteric fruit. Ethylene biosynthesis has been studied clearly and ACC synthase (ACS) is the rate-limited enzyme. ACO (ACC oxidase) is another important enzyme in ethylene biosynthesis. By exploring the pear genome, we identified 13 ACS genes and 11 ACO genes, respectively, and their expression patterns in fruit and other organs were investigated. Among these genes, 11 ACS and 8ACO genes were expressed in pear fruits. What’s more, 4 ACS and 3ACO genes could be induced by Ethephon and inhibited by 1-MCP treatment. This study is the first time to explore ACS and ACO genes at genome-wide level and will provide new data for research on pear fruit ripening.

     

Pyridinylpyrimidines selectively inhibit human methionine aminopeptidase-1

Cellular protein synthesis is initiated with methionine in eukaryotes with few exceptions. Methionine aminopeptidases (MetAPs) which catalyze the process of N-terminal methionine excision are essential for all organisms. In mammals, type 2 MetAP (MetAP2) is known to be important for angiogenesis, while type 1 MetAP (MetAP1) has been shown to play a pivotal role in cell proliferation. Our previous high-throughput screening of a commercial compound library uncovered a novel class of inhibitors for both human MetAP1 (HsMetAP1) and human MetAP2 (HsMetAP2). This class of inhibitors contains a pyridinylpyrimidine core. To understand the structure–activity relationship (SAR) and to search for analogues of 2 with greater potency and higher HsMetAP1-selectivity, a total of 58 analogues were acquired through either commercial source or by in-house synthesis and their inhibitory activities against HsMetAP1 and HsMetAP2 were determined. Through this systematic medicinal chemistry analysis, we have identified (1) 5-chloro-6-methyl-2-pyridin-2-ylpyrimidine as the minimum element for the inhibition of HsMetAP1; (2) 5′-chloro as the favored substituent on the pyridine ring for the enhanced potency against HsMetAP1; and (3) long C4 side chains as the essentials for higher HsMetAP1-selectivity. At the end of our SAR campaign, 25b, 25c, 26d and 30a–30c are among the most selective and potent inhibitors of purified HsMetAP1 reported to date. In addition, we also performed crystallographic analysis of one representative inhibitor (26d) in complex with N-terminally truncated HsMetAP1.     

Molecular dynamics simulation on the allosteric analysis of the c‐di‐GMP class I riboswitch induced by ligand binding

Riboswitches are RNA molecules that regulate gene expression using conformation change, affected by binding of small molecule ligands. Although a number of ligand‐bound aptamer complex structures have been solved, it is important to know ligand‐free conformations of the aptamers in order to understand the mechanism of specific binding by ligands. In this paper, we use dynamics simulations on a series of models to characterize the ligand‐free and ligand‐bound aptamer domain of the c‐di‐GMP class I (GEMM‐I) riboswitch. The results revealed that the ligand‐free aptamer has a stable state with a folded P2 and P3 helix, an unfolded P1 helix and open binding pocket. The first Mg ions binding to the aptamer is structurally favorable for the successive c‐di‐GMP binding. The P1 helix forms when c‐di‐GMP is successive bound. Three key junctions J1/2, J2/3 and J1/3 in the GEMM‐I riboswitch contributing to the formation of P1 helix have been found. The binding of the c‐di‐GMP ligand to the GEMM‐I riboswitch induces the riboswitch's regulation through the direct allosteric communication network in GEMM‐I riboswitch from the c‐di‐GMP binding sites in the J1/2 and J1/3 junctions to the P1 helix, the indirect ones from those in the J2/3 and P2 communicating to P1 helix via the J1/2 and J1/3 media.     

外源木聚糖酶在枯草芽胞杆菌中信号肽筛选

[目的]本试验旨在筛选引导表达外源木聚糖酶基因高效分泌的信号肽,为枯草芽胞杆菌木聚糖酶高效分泌表达系统提供元件.[方法]构建信号肽筛选载体,载体是以含壮观霉素抗性基因的大肠-枯草穿梭载体为基本骨架,目标蛋白为耐碱性木聚糖酶,可在麦芽糖启动子Pglv诱导下表达.从枯草芽胞杆菌A1747基因组中扩增获得24个Sec途径信号肽,并将其全部链接到至筛选载体上,并在枯草芽胞杆菌WB700中实现表达分泌.重组菌在3%麦芽糖诱导下培养24h后用DNS法测定上清酶活.[结果]成功构建信号肽筛选载体pGPSX及24个表达载体,实现木聚糖酶表达分泌.且不同信号肽对于引导外源木聚糖酶分泌能力不同,其中YnfF信号肽引导分泌目标蛋白效率最高,上清酶活为37.2IU/mL.[结论]试验证明在枯草杆菌中对外源蛋白进行信号肽筛选是提高其分泌的有效途径,并获得了针对木聚糖酶高效分泌信号肽YnfF.     

石灰石粉施用剂量对重庆酸雨区受害 马尾松林细根生长的影响

马尾松(Pinus massoniana)为重庆的重要乡土树种和主要造林树种,担负着特殊而重要的生态环境保全功能,但在酸雨的长期影响下,其健康状况持续低下。为探究不同剂量石灰石粉对马尾松细根生长的影响,给受害马尾松纯林的恢复与管理提供科学依据,2004年5月在属于酸雨区的重庆市江北区铁山坪林场,采用随机区组设计,设立在马尾松纯林酸化土壤表面一次性撒施石灰石粉0(不撒施对照)、1、2、3,4t/hm² 5个处理的长期定位观测试验。2006年10月的监测结果表明,与对照相比,撒施石灰石粉可明显促进腐殖质层和0-20、20-40、40-60cm土层马尾松细根的生长(P < 0.05),尤其是改善腐殖质层和20-40、40-60cm的根量,在腐殖质层和0-60cm马尾松细根的干重总密度、长度总密度、表面积总密度、体积总密度和根尖总密度与对照各项密度指标对应值的比值,1t/hm²下分别为1.53、1.34、1.39、1.24,1.48,2t/hm²下分别为2.05、1.83、1.94、1.79,2.04,3t/hm²下分别为2.17、2.02、2.02、1.78,2.32,4t/hm²下分别为2.68、2.52、2.62、3.70和2.71,改善效果总体上随撒施剂量增大而增强。从有限时间(2-3a)内观测的根系生长改善效果与经济投入的比值来看,建议一次性撒施的最佳剂量为2t/hm²,但在考虑时间加长情况下,效果最好和最经济的剂量还有待后续研究。     

拟南芥ABA敏感突变体asm1的分离与鉴定

以拟南芥(Arabidopsis thaliana)为研究材料,从T-DNA突变体库中筛选分离得到1株脱落酸(ABA)敏感突变体asm1(ABA sensitive mutant 1,asm1),在含有ABA的培养基中,与野生型相比,asm1突变体的根伸长明显受到抑制,且其种子萌发结果显示asm1对ABA同样表现出敏感特性。在生长发育方面,asm1突变体抽苔时间提前,植株矮化,并且荚果长度明显小于野生型。利用远红外成像系统分析发现,在干旱胁迫下asm1突变体叶面温度高于野生型;失水率分析显示突变体失水率降低以及水分散失减少。遗传学分析表明,asm1是单基因隐性突变且与一个T-DNA插入共分离;通过图位克隆成功获得候选基因ASM1。RT-PCR结果显示,在突变体中ASM1的表达受到抑制,并且能够调控多种ABA信号通路和胁迫应答基因的表达水平。研究结果表明,ASM1可能参与调控ABA信号转导并应答干旱胁迫。     

非洲猪瘟病毒I226R蛋白抑制cGAS-STING通路介导的天然免疫应答

本研究旨在探究非洲猪瘟病毒(African swine fever virus, ASFV) I226R蛋白(I226R protein, pI226R)抑制cGAS-STING信号通路的作用机制。利用双荧光素酶报告系统和实时荧光定量PCR (real-time quantitative PCR, qPCR)证明pI226R显著抑制cGAS-STING通路介导的I型干扰素及干扰素刺激相关基因的产生。免疫共沉淀及激光共聚焦显微镜试验发现pI226R与cGAS蛋白相互作用。免疫印迹分析证明pI226R通过自噬-溶酶体途径促进cGAS蛋白的降解。同时,pI226R阻碍了cGAS与E3泛素连接酶三基序蛋白56 (tripartite motif protein 56, TRIM56)的结合,导致cGAS的单泛素化减弱,从而抑制了cGAS的活化和cGAS-STING通路的激活。总之,本研究证明ASFV pI226R通过拮抗cGAS进而抑制宿主的抗病毒天然免疫反应,进一步增加了对研究ASFV免疫逃逸机制的理解,为疫苗的研发提供了理论基础。     

六盘山典型森林伴随降水的总有机碳(TOC)通量变化特征

在六盘山香水河小流域,选择6种典型森林样地,测定了2011年生长季的大气降水、穿透水、干流、枯落物渗漏水和主根系层(0—30 cm深)土壤渗漏水的总有机碳(TOC)浓度及其相应的通量变化。结果表明,在降水转化为由穿透雨和干流组成的林下降水中,所有样地的TOC浓度都不同程度地增大;虽然林冠截持使林下降水减小,但因雨水淋洗和与林冠发生碳交换,各样地林下降水携带的生长季TOC通量(kg/hm2)(华北落叶松人工林132.28、华山松次生林106.56、油松人工林94.10、灌木林79.49、桦木林66.52、辽东栎次生林63.01)都比林外降水(53.17)不同程度地明显增大,整体看来,林冠的TOC淋出作用在针叶林很大,在阔叶林较弱。在6种森林样地的枯落物层渗漏水中,其TOC浓度彼此相差不大,平均为24.51 mg/L,高于林冠穿透水的TOC浓度;受枯落物截持部分降水及与枯落物TOC交换的影响,4个样地枯落物渗漏水的TOC通量(kg/hm2)(桦木次生林84.35、野李子灌丛129.35、辽东栎次生林79.21、油松人工林114.93)都比其林下降水TOC通量增加了,但华北落叶松人工林和华山松次生林的TOC通量分别降至90.76和104.90 kg/hm2。在测定的华北落叶松人工林和华山松次生林的主根系层(0—30 cm)土壤渗漏水中,TOC浓度均低于枯落物渗漏水;由于水量减小和与土壤发生碳交换,土壤渗漏水的TOC通量均显著低于枯落物渗漏水,两个林分样地分别降至43.04和66.33 kg/hm2。整体来看,林外降水携带的TOC输入通量在林地TOC输入中占有重要地位,林冠的TOC淋洗使其程度不同地增加TOC通量,枯落物层具有增加或减少TOC通量的作用,但主根系层土壤会显著减少TOC输出通量,所以是固定TOC的重要场所。     

Molecular Cytogenetic Identification of a New Wheat-Rye 6R Chromosome Disomic Addition Line with Powdery Mildew Resistance

Rye (Secale cereale L.) possesses many valuable genes that can be used for improving disease resistance, yield and environment adaptation of wheat (Triticum aestivum L.). However, the documented resistance stocks derived from rye is faced severe challenge due to the variation of virulent isolates in the pathogen populations. Therefore, it is necessary to develop desirable germplasm and search for novel resistance gene sources against constantly accumulated variation of the virulent isolates. In the present study, a new wheat-rye line designated as WR49-1 was produced through distant hybridization and chromosome engineering protocols between common wheat cultivar Xiaoyan 6 and rye cultivar German White. Using sequential GISH (genomic in situ hybridization), mc-FISH (multicolor fluorescence in situ hybridization), mc-GISH (multicolor GISH) and EST (expressed sequence tag)-based marker analysis, WR49-1 was proved to be a new wheat-rye 6R disomic addition line. As expected, WR49-1 showed high levels of resistance to wheat powdery mildew (Blumeria graminis f. sp. tritici, Bgt) pathogens prevalent in China at the adult growth stage and 19 of 23 Bgt isolates tested at the seedling stage. According to its reaction pattern to different Bgt isolates, WR49-1 may possess new resistance gene(s) for powdery mildew, which differed from the documented powdery mildew gene, including Pm20 on chromosome arm 6RL of rye. Additionally, WR49-1 was cytologically stable, had improved agronomic characteristics and therefore could serve as an important bridge for wheat breeding and chromosome engineering.