Tup1基因缺失对酿酒酵母耐高糖性状的影响

研究Tup1基因对酿酒酵母（Saccharomyces cerevisiae）细胞耐高糖性状的影响。利用Cre/loxP系统对单倍体细胞中的转录因子Tup1基因进行敲除,单倍体细胞复倍后研究基因缺失菌株的性状变化。结果表明,Tup1基因缺失虽然减弱了菌株的呼吸能力,但却增强了菌株在高浓度葡萄糖培养基中的生长和发酵能力。Tup1基因可能通过调节酿酒酵母细胞呼吸强度来调控细胞的高糖应激反应。     

Identification of a bi-directional promoter from Tomato yellow leaf curl China virus

A bi-directional promoter of Tomato yellow leaf curl China virus (TYLCCNV) was obtained with the total DNA from TYLCCNV isolate Y10 infected tobacco leaves as a template. Plant expression vectors were constructed by fusing the amplified DNA fragment with the gus gene and nopaline terminator in different orientations. The vectors containing promoter fragments were transferred into leaf cells and plant stems of Nicotiana benthamiana by Agrobacterium-mediated method. Transient expression results showed that both the complementary and virion-sense promoters could drive the gus gene to express, and the GUS activity of the complementary-sense promoter was stronger than that of the virion-sense. Co-expression of the vector containing βC1 gene of TYLCCNV DNAβ with the vector containing a bi-directional promoter revealed that the βC1 protein has no impact on expression of either the virion- or the complementary-sense promoter.     

Identification of a bi-directional promoter from Tomato yellow leaf curl China virus

A bi-directional promoter of Tomato yellow leaf curl China virus (TYLCCNV) was obtained with the total DNA from TYLCCNV isolate Y10 infected tobacco leaves as a template. Plant expression vectors were constructed by fusing the amplified DNA fragment with the gus gene and nopaline terminator in different orientations. The vectors containing promoter fragments were transferred into leaf cells and plant stems of Nicotiana benthamiana by Agrobacterium-mediated method. Transient expression results showed that both the complementary and virion-sense promoters could drive the gus gene to express, and the GUS activity of the complementary-sense promoter was stronger than that of the virion-sense. Co-expression of the vector containing βC1 gene of TYLCCNV DNAβ with the vector containing a bi-directional promoter revealed that the βC1 protein has no impact on expression of either the virion- or the complementary-sense promoter.     

Expression of ipt gene driven by tomato fruit specific promoter and its effects on fruit development of tomato

Fruit specific promoter (2A12) from Lycopersicom esculentum and cDNA of isopentenyl-transferase (ipt) from Ti plasmid of Agrobacterium tumerfaciens C58 were cloned by PCR procedure respectively. Two plant expression vectors with 2A12/gus or 2A12/ipt were respectively constructed. These two chimeric genes were transferred into tomato by Agrobacterium mediated procedure. The results of Southern hybridization showed that the fusion genes had been integrated into tomatoes. The result of gus histochemical staining showed that 2A12 had high fruit specific expressive capability in transgenic tomato. The ipt expression resulted in accumulation of high level of cytokinins (CTKs) in fruit lead to developmental changes in fruits and seeds. The fruit of ipt transformed tomato had the hyperplastic placenta with very few seeds or even seedless. The shelf life of transgenic fruits elongated for 1–2 weeks. The ratio of fruit set, the dry weight of fruit and the crude protein content in fruit were increased, while the soluble sugar of fruits decreased.     

核桃JrEFM1转录因子响应逆境及激素的表达模式

MYB转录因子在调控植物生长发育和逆境响应方面发挥着重要的作用.通过克隆获得了核桃的1条R1-MYB类转录因子EFM基因(命名为JrEFM1),利用生物信息学和实时荧光定量RT-PCR(RT-qPCR)技术,分析在不同非生物及植物激素处理下JrEFM1的表达规律,探究了JrEFM1的基本生物学功能.结果显示,JrEFM1的编码区长为1 320 bp,编码蛋白含439个氨基酸,分子量为48.322 KDa,理论等电点为9.26.与葡萄、番薯等具有较近的进化关系.其启动子包含干旱胁迫(MBS)、热激响应(HSE)、水杨酸(SA)、玉米素(O2-site)和赤霉素响应(GARE-motif)等相关元件.对JrEFM1在干旱,冷害,热激,ABA,JA,SA处理下的表达情况进行分析,发现JrEFM1可被这些处理不同程度地诱导,同时根和叶表现出不同的转录水平.表明JrEFM1可响应逆境胁迫,并与激素信号通路相关;JrEFM1可作为核桃逆境响应机制研究及抗逆育种的优良候选基因.     

Overexpression of rice OsLOL2 gene confers disease resistance in tobacco to Pseudomonas syringae pv. Tabaci

LSD1-related proteins of Arabidopsis with LSDl-like zinc finger domains regulate disease resistance and programmed cell death （PCD）. We cloned a rice OsLOL2 gene, orthologous to LSD1 of Arabidopsis and expressed it in a tobacco plant. Transgenic tobacco lines displayed enhanced disease resistance to a virulent bacterium Pseudomonas syringae pv. tabaci （Pst）. RT-PCR analysis showed that overexpression of OsLOL2 in transgenic tobacco lines resulted in upregulation of two pathogenesis-related （PR） protein genes, PR2 and PR5. Our results suggest that overexpression of OsLOL2 in transgenic tobacco enhances the resistance through the induction of PR proteins and hypersensitive response-like reaction.     

银杏疫病的研究

对银杏苗圃中发生苗枯的植株进行病原分离及回接试验,发现引起银杏苗枯的病原为Ｐｈｙｔｏｐｈｔｈｏｒａｓｐ．称这种苗枯为银杏疫病。该病危害银杏幼嫩苗木的地上部分,未见危害根部和多年生成林树。叶片受害似开水烫伤状,茎干受害则变黑,黑色病斑环绕茎干一周后,病茎以上部分叶片青枯萎垂。病原菌在ＰＳＡ培养基上,菌落白色绒毛状,２８℃下７ｄ可长出大量孢子囊,易产生厚壁孢子,病菌生长适宜温度为２４℃～３２℃。人工接种可致受伤银杏叶片及幼嫩茎干严重发病,但无伤茎叶发病较轻或不受害。在田间,该病５月初开始发生,３ｄ～５ｄ病情达到高峰,苗圃的发病率为１０％～２０％,有明显的发病中心,５月底病情稳定,以后病害不再发生。用含药培养基及含药纸碟法测定,甲霜灵及甲霜灵锰锌对病原菌有很强的抑制作用。病原种鉴定正在进行。     

温度和光照对克氏原螯虾生物钟基因PcClk与PcCry1表达的影响

生物钟基因clock(Clk)和cryptochrome(Cry)参与生物内在节律调控。克氏原螯虾(Procambarus clarbii)是我国重要的水产养殖经济虾类,为探究温度、光照对克氏原螯虾生长及其生物钟基因PcClk与PcCry1节律振荡表达的影响,采用实时定量PCR技术探讨克氏原螯虾PcClk和PcCry1基因在脑、眼柄和肝胰腺组织中的mRNA表达模式,并探讨PcClk和PcCry1基因在温度和光照周期适应过程中是否发生适应性进化。荧光定量PCR检测结果显示,克氏原螯虾的PcClk基因和PcCry1基因在脑、眼柄和肝胰腺中均有表达。在不同光照周期培养下,PcClk基因和PcCry1基因在脑、眼柄中的mRNA表达有一定的振荡节律,在25℃、12 LD的培养模式下,脑中PcClk基因和PcCry1基因mRNA表达的节律性会更明显。选择压力分析显示,经PAML和Datamonkey共同筛选出的PcCLK正选择位点为71与213,PcCRY1正选择位点为63。以上结果表明,克氏原螯虾生物钟基因PcClk和PcCry1受到了选择压力,克氏原螯虾PcClk基因和PcCry1基因是内源性的节律基因。     

番茄褪绿病毒及植株对其侵染响应性研究进展

目前全球已知的番茄褪绿病毒(Tomato chlorosis virus,ToCV)分离株系可分成3组,它们在中国境内都有被发现,但仍呈区域性分布.ToCV病害主要通过阻断粉虱传播来防治,更长期有效的防控则基于对植株对ToCV侵染的敏感性和抗性的分子遗传基础的了解,和对病毒具有耐受或者抗性的作物品种的培育.为此,综述了...     

The concentration quenching characteristics of 5D3-7FJ and 5D4-7FJ (J=0―6) transitions of Tb3+ in YBO3:Tb3+ phosphor

The photoluminescence quenching behaviors of ^5D3-^7Fj and ^5D4-^7Fj （J = 0—6） transitions of Tb^3＋ in YBO3：Tb under 130—290 nm excitation were systematically investigated. The results revealed that the quenching concentrations of both ^5D3-^7Fj and ^5D4-^7Fj transitions of Tb^3＋ in YBO3：Tb were mainly dependent on excitation wavelength. Particularly, the quenching concentrations of ^5D4-^7Fj transitions of Tb^3＋ under 130—290 nm excitation were correlated with excitation bands of YBO3：Tb. The quenching concentrations of ^5D3-^7Fj transitions remained at low concentration （2%） under 186—290 nm excitation and then increased gradually with energy of incoming excitation photon when excited at 130—186 nm. This dependence should be involved in their excitation mechanisms and quenching pathway in particular excitation region.[第一段]     

Magnetoelectric and converse magnetoelectric res ponses in TbxDy1-xFe2-y alloy ＆ Pb（Mg1/3Nb2/3）（1-x）TixO3 crystal laminated composites

Measured results of magnetoelectric （ME） and converse magnetoelectric （CME） effects of TbxDy1-xFe2-y/ Pb（Mg1/3Nb2/3）（1-x）TixO3/TbxDy1-xFe2-y （TD/PMNT/TD） and PMNT/TD/PMNT laminated composites are presented. ME effect was determined by measuring laminate voltage output under a Helmholtz-generated AC field biased by a DC field （0-1 kOe） （1Oe = 79.58 A/m）. The CME effect was measured by recording the voltage induced in a solenoid encompassing the ME sample while exposed to a DC bias field and PMNT layer driven by a 10 V AC source. The ME and CME responses in the two laminated structure are linear. The highest values of ME coefficients in TD/PMNT/TD and PMNT/TD/PMNT composites are 384 mV/Oe and 158 mV/Oe, respectively, while the highest values of CME coefficients in the two composites are 118 mG/V and 162 mG/V （1 G=10^-4 T）, respectively.     

黑曲霉木聚糖酶基因xynB的克隆及在酿酒酵母中的表达

从黑曲霉(Aspergillus niger)克隆木聚糖酶基因xynB,利用重叠延伸PCR法去除其中的内含子.将该基因与质粒pYES2连接,构建真核表达载体,用醋酸锂化转法导人酿酒酵母(Sacharom yces cerevisiae)INVSC1菌株表达.利用α信号肽替换基因原信号肽,以考察信号肽对表达蛋白分泌的影响.结果获得2株重组菌株,分别为xynB连接原信号肽的pYES2-xynB菌株和xynB连接α信号肽的pYES2-xynB-α菌株.木聚糖酶B成功表达,SDS-PAGE检测到约24kDa目的蛋白质条带.木聚糖酶B最适催化温度为50℃,最适催化pH值为5.0,Mn2+对酶活具有强烈的抑制作用.将α信号肽取代原信号肽使酶活达到最高的诱导时间由72h缩短为48h,但是置换信号肽使最高酶活由7.59U降低为3.97U.     

Fine mapping of the red plant gene R1 in upland cotton(Gossypium hirsutum)

Sub 16 is a substitution line with G. hirsutum cv. TM-1 genetic background except that the 16th chromosome (Chr. 16) is replaced by the corresponding homozygous chromosome of G. barbadense cv. 3-79, and T586 is a G. hirsutum multiple gene marker line with 8 dominant mutation genes. The R ₁ gene for anthocyanin pigmentation was tagged in Chr. 16 in T586. The objective of this research was to screen SSR markers tightly linked with R ₁ by using the F₂ segregating population containing 1259 plants derived from the cross of Sub 16 and T586 and the backbone genetic linkage map from G. hirsutum×G. barbadense BC₁ newly updated by our laboratory. Genetic analysis suggested that the segregation ratio of red plants in the F₂ population fit Mendelian 1:2:1 inheritance, confirming that the red plant trait was controlled by an incomplete dominance gene. Preliminary mapping of R ₁ was conducted using 237 randomLy selected F₂ individuals and JoinMap v3.0 software. Then, a fine map of R1 was constructed using the F₂ segregating population containing 1259 plants, and R ₁ was located between NAU4956 and NAU6752, with only 0.49 cM to the nearest maker loci (NAU6752). These results provided a foundation for map-based cloning of R ₁ and further development of cotton cultivars with red fibers by transgenic technology. Supported by National Natural Science Foundation of China (Grant No. 30730067) and Programme of Introducing Talents of Discipline to Universities (Grant No. B08025)     

Speciation of the genus Arthricocephalus Bergeron, 1899 (Trilobita) from the late Early Cambrian and its stratigraphic significance

The genus Arthricocephalus Bergeron, 1899 is revised, and Halipanktos Balker & Peel, 1997 is suggested here as a senior synonym. The subgenus Arthricocephalus (Arthricocephalites) Chien & Lin in Lu et al., 1974 is considered as a separate genus. Of the 20 previously assigned species of Arthricocephalus (Arthricocephalus) Bergeron, 1899, Arthricocephalus (Arthricocephalites) Chien & Lin in Lu et al., 1974, Arthricocephalus (Euarthricocephalus) Ju, 1983 are lumped into eight species. The speciation trend of Arthricocephalus and Arthricocephalites is demonstrated based on their stratigraphic occurrences. It not only enhances the resolution of the biostatigraphic zonation in the uppermost Lower Cambrian, but also represents a potential candidate to define the Duyunian stage. The base of the stage is suggested at the first appearance datum (FAD) of Arthricocephalus chauveaui Bergeron, 1899 within the evolutionary lineage from Ar. jiangkouensis Yin in Yin & Li,1978 to Ar. chauveaui in a global scale.     

Efficient transformation and expression of gfp gene in the edible mushroom Pleurotus nebrodensis

An efficient transformation method mediated by PEG-protoplasts was developed for the newly commercial edible mushroom Pleurotus nebrodensis. Two plasmids were used to co-transform protoplasts of P. nebrodensis. One plasmid is pAN7-1 containing a positive selectable marker gene hph conferring hygromycin B resistance. Another plasmid is pBlue-GFP containing a reporter gene gfp conferring green fluorescent protein. PCR and Southern blot analysis showed that hph gene or/and gfp gene were integrated into the genome of P. nebrodensis transformants. The transformation efficiency of the positive selectable marker gene hph was 3 transformants per microgram of plasmid pAN7-1 DNA, which was about 30 times higher than that previously reported in thoroughly studied Pleurotus species such as Pleurotus ostreatus. The transformation efficiency of the reporter gene gfp was 9 transformants per microgram of plasmid pBlu-GFP DNA. The co-transformation efficiency was 23.68%. This is the first report that a ＂reporter＂ gene, green fluorescent protein gene can be successfully stably exoressed in this Pleurotus species.     

Speciation of the genus Arthricocephalus Bergeron, 1899 (Trilobita) from the late Early Cambrian and its stratigraphic significance

The genus Arthricocephalus Bergeron, 1899 is revised, and Halipanktos Balker & Peel, 1997 is suggested here as a senior synonym. The subgenus Arthricocephalus (Arthricocephalites) Chien & Lin in Lu et al., 1974 is considered as a separate genus. Of the 20 previously assigned species of Arthricocephalus (Arthricocephalus) Bergeron, 1899, Arthricocephalus (Arthricocephalites) Chien & Lin in Lu et al., 1974, Arthricocephalus (Euarthricocephalus) Ju, 1983 are lumped into eight species. The speciation trend of Arthricocephalus and Arthricocephalites is demonstrated based on their stratigraphic occurrences. It not only enhances the resolution of the biostatigraphic zonation in the uppermost Lower Cambrian, but also represents a potential candidate to define the Duyunian stage. The base of the stage is suggested at the first appearance datum (FAD) of Arthricocephalus chauveaui Bergeron, 1899 within the evolutionary lineage from Ar. jiangkouensis Yin in Yin & Li,1978 to Ar. chauveaui in a global scale.     

不同产地独活及其混伪品的质量差异性分析

多点采样,对不同来源的独活及其混伪品进行质量差异性分析.测定不同产地独活及其混伪品总灰分、酸不溶性灰分、蛇床子素和二氢欧山芹醇当归酸酯质量分数,并结合HPLC指纹图谱对不同产地独活及其混伪品开展质量评价.结果表明,不同产地独活除陕西样品的蛇床子素略低于《中华人民共和国药典》(以下简称《药典》)标准外,其余样品均符合《药典》质量要求,而独活混伪品的蛇床子素质量分数均远低于正品独活质量分数.不同产地的独活HPLC指纹图谱有25个共有峰,各独活样品的指纹图谱与对照图谱的相似度在0.9以上,所建立的指纹图谱可用于独活的质量评价.因此,多指标的测定结果结合HPLC指纹图谱,可有效对不同产地的独活质量进行差异性分析和评价;混伪品蛇床子素的质量分数远低于正品独活,蛇床子素的质量分数高低可作为正品独活区别于混伪品的一个重要特征指标.