Hormonal Regulation of Ca2+ Stimulated K+ Influx and Ca2+, K+- ATPase in Rice Roots: in vivo and in vitro Effects of Auxins and Reconstitution of the ATPase

The role of natural and synthetic auxins in regulation of ion transport and ATPase activity was studied in rice roots (Oryza sativa L. cv. Dunghan Shah). In vivo treatment of seedlings with 2,4-dichlorophenoxyacetic acid at 2 × 10^?6M for a short period enhanced subsequent Ca²⁺ stimulated K⁺ influx and ATPase activity, while a longer treatment diminished both K⁺ influx and ATPase activity. Indoleacetic acid at 10^?10–10^?8M induced ATPase activity. In in vitro experiments both 2,4-dichloro phenoxyacetic acid and indoleacetic acid (10^?10–10^?8M) stimulated Ca²⁺, K⁺-ATPase activity of a plasmalemma rich micro somal fraction from the roots. Acetone extracted ATPase preparations lost their activity. The enzyme regained its activity and its sensitivity towards ions (Ca²⁺+ K⁺) when reconstituted with phosphatidyl choline. Addition of auxins also indicated that the presence of the lipid was necessary in the interaction between the ATPase and auxins. Auxins and ions probably interact with the intact ATPase lipoprotein complex, which may possess a receptor site for the auxins, possibly as a sub unit.     

Introduction of a Micrococcus plasmid in Escherichia coli

A 6-MDa plasmid (pMQV10), carrying cholesterol hydroxylase and streptomycin-resistance genes, from a gram-positive strain of Micrococcus sps., (RJ6) has been successfully transformed in gram-negative Escherichia coli K12 C600. pMQV10 is maintained stably and expresses its drug resistance in the new host.     

Extrachromosomal DNA in Bacillus thuringiensis var. kurstaki, var. finitimus, var. sotto, and in Bacillus popilliae

Four entomopathogenic bacteria contained extrachromosomal deoxyribonucleic acid (DNA) molecules of various sizes. Bacillus thuringiensis var. kurstaki contained twelve elements banding on agarose gels that ranged from 0.74 to > 50 × 10⁶ daltons, three of which were giant extrachromosomal DNA elements. B. thuringiensis var. sotto contained one giant extrachromosomal DNA element with a molecular size of about 23.5 × 10⁶ daltons and two lesser elements of 0.80 and 0.62 × 10⁶ daltons. B. thuringiensis var. finitimus harbored two giant DNA elements corresponding to >50 × 10⁶ daltons and two lesser bands with relative small size (0.98 and 0.97 × 10⁶ daltons). B. popilliae contained no giant extrachromosomal DNA elements but did contain two smaller elements corresponding to 4.45 and 0.58 × 10⁶ daltons. The possible use of extrachromosomal DNA elements that prove to be autonomous replicons for recombinant DNA studies is discussed.     

Apelin activates L-arginine/nitric oxide synthase/nitric oxide pathway in rat aortas

Apelin was recently found to be an inotropic polypeptide in isolated rat hearts, and intravenous injection of apelin can induce a transient decrease in blood pressure. To illustrate the mechanism of apelin-induced vasodilation, we observed the in vitro effects of apelin on the L-arginine (L-Arg)/nitric oxide (NO) pathway in the incubated, isolated rat aorta. Apelin stimulated vascular NO(2)(-) product and NOS activation in a concentration- and time-dependent manner. Compared with no apelin treatment, incubation with apelin (10(-9), 10(-8), and 10(-7)mol/L) increased NO(2)(-) product by 33%, 46%, and 69% (all p<0.01), respectively, and Ca(2+)-dependent constitutive NOS (cNOS) activity by 200%, 460%, and 550% (all p<0.01), respectively. However, Ca(2+)-independent NOS (iNOS) activity was not significantly altered (p>0.05). Apelin incubation (10(-9), 10(-8), and 10(-7)mol/L) increased L-Arg uptake by 130%, 180%, and 240% (all p<0.01), respectively. The mRNA level of cationic amino acid transporters, CAT-1 and CAT-2B, in rat aortic tissues treated with 10(-7)mol/L apelin was increased by 110% and 128%, respectively (both p<0.01). Incubation with 10(-7)mol/L apelin elevated eNOS mRNA and protein levels, by 53% (p<0.05) and 319% (p<0.01), respectively. Collectively, these results demonstrate that apelin directly activated the vascular L-Arg/NOS/NO pathway, which could be one of the important mechanisms of apelin-regulated vascular function.     

Starch-sugar interconversion in Solanum tuberosum

The changes in starch, sugars, and respiration of both immature and mature potato tubers (variety King Edward) caused by transfer from +10° to +2° and back to +10°, were followed throughout. At each storage temperature the tubers were allowed to reach a steady state before transfer to another temperature. In potatoes transferred from +10° to +2°, the sugar at first rose rapidly and then reached a constant value after 30 days. The respiration showed a characteristic pattern, for the first 5–8 days being below the initial value, then rising to a maximum at 14 days and finally returning to the initial value at 28 days. In potatoes transferred from +2° to +10° the sugar declined steadily, the respiration reaching a maximum after 10 days and then slowly falling to a value slightly above the initial value. Quantitative analysis of the results showed that the sum of starch + sugar + CO₂ expressed in equivalent anhydrohexose units did not change throughout the various changes in temperature. This work provided a quantitative experimental basis for what had hitherto been an assumption. Starch was the only polysaccharide involved in these carbohydrate changes. No change in the amylose/amylopectin ratio was detected either during the breakdown of starch (temperature change +10° to +2°) or during its synthesis (+2° to + 10°). The increased respiration which accompanied any change in temperature was related quantitatively to the formation of sucrose from starch (+10° to +2°) and starch from sugar (+2° to + 10°). The ATP equivalent of the extra CO₂ output was of the same order as that predicted on the basis of known biochemical pathways linking starch and sugar.     

草莓生长素响应因子FvARF10和FvARF18基因的克隆及表达分析

该研究以森林草莓(Fragaria vesca)为试验材料,利用RT-PCR技术克隆草莓FvARF10、FvARF18基因全长cDNA序列。生物信息学分析表明,FvARF10、FvARF18基因开放阅读框(ORF)分别为2 056bp和2 100bp,编码685和699个氨基酸,预测其分子量和等电点分别为75.73、76.78kD和8.12、6.55。系统进化树分析表明,FvARF10与苎麻BnARF10聚为一个分支,FvARF18与桑树MnARF18聚集在一个分支上,进化关系较密切。亚细胞定位预测分析表明,FvARF10、FvARF18蛋白均定位于细胞核或细胞质中。启动子分析显示,FvARF10、FvARF18基因均具有多样化的激素应答元件。荧光定量PCR结果显示,FvARF10和FvARF18基因在草莓不同组织中均有表达,FvARF10在果实中表达较高,且在半熟果中表达量最高,而FvARF18在茎中相对表达量最高;外源激素IAA处理1h后,FvARF10和FvARF18基因受到强烈诱导均呈上调表达,而后渐恢复正常水平。研究结果表明,FvARF10和FvARF18基因可能参与草莓的营养生殖及果实的生长发育调控,且与生长素信号转导过程相关。     

Transduction in Rhizobium meliloti

Summary Of 21Rhizobium meliloti temperate phages, 12 transduced streptomycin resistance (str-r) with a frequency of 10⁻⁵ to 10⁻⁷. Lysine dependence was transduced with frequency of 10⁻⁷. Five transducing phages were used in experiments on the transfer of effectiveness from effective donor strain to 11 ineffective strains. However, plant tests did not reveal changes in recipient effectiveness.     

Trypanosoma musculi infections in complement-deficient mice

Strains of mice (CFW, C57B1/10Sn, B10.D2/nSn, and B10.D2/oSn) were infected with Trypanosoma musculi (Trypanosoma duttoni). The complement-deficient B10.D2/oSn mice showed typical parasitemias similar to those presented by the strains possessing hemolytic complement activity. Peak parasitemias occurred 12 days postinoculation. The highest parasitemias were measured in CFW mice (657 ± 82 T. musculi/30 hi-power fields), while infections in C56BL/10Sn (528 ±44 T. musculi/30 h.p.f.), B10.D2/oSn (502 ± 20 T. musculi/30 h.p.f.) and B10.D2/nSn (512 ± 35 T. musculi/30 h.p.f.) were less severe and quantitatively comparable. The percentages of dividing forms were similar during infections in each of the strains. While parasites were detected in peripheral blood until Day 22 of infection in three of 10 C57BL/10Sn mice, none could be found at this time in blood films of CFW, B10.D2/nSn or B10.D2/oSn mice. Giemsa stained kidney imprints indicated the presence of parasites in animals of each of the strains after 33 days, when trypanosomes could no longer be detected in the peripheral blood of the mice. The minor variations in the parasitemias appeared related to the mouse strain. Complement dependent, antibody mediated immune cytolysis was not indicated as a mechanism for the elimination of T. musculi by the infected mouse.     

Models for the 3(10)-helix/coil,pi-helix/coil,and alpha-helix/3(10)-helix/coil transitions in isolated peptides.

Models for the 3(10)-helix/coil and pi-helix/coil equilibria have been derived. The theory is based on classifying residues into helical or nonhelical (coil) conformations. Statistical weights are assigned to residues in a helical conformation with an associated helical hydrogen bond, a helical conformation with no hydrogen bond, an N-cap position, a C-cap position, or the reference coil conformation. The models for alpha-helix formation and 3(10)-helix formation have also been combined to describe a three-state equilibrium in which alpha-helical, 3(10)-helical, and coil conformations are populated. The results are compared with the modified Lifson-Roig theory for the alpha-helix/coil equilibrium. The comparison accounts for the experimental observations that 3(10)-helices tend to be short and pi-helices are not favored for any length. This work may provide a framework for quantitatively rationalizing experimental work on isolated 3(10)-helices and mixed 3(10)-/alpha-helices.     

巨桉扩展蛋白EgrEXPA8和EgrEXPA10基因的克隆和表达特性分析

为探讨扩展蛋白在桉树生长发育中的作用,以在桉树初生生长到次生生长转换转录组测序中筛选出的差异表达基因EgrEXPA8和EgrEXPA10为基础,从巨桉(Eucalyptusgrandis)中克隆了2个扩展蛋白基因EgrEXPA8和EgrEXPA10,分别编码249和244个氨基酸,属于亲水蛋白,但Egr EXPA8稳定性高于Egr EXPA10。q RT-PCR分析表明,Egr EXPA8和Egr EXPA10基因均在幼叶和茎尖组织中表达量较高,在木质部和韧皮部表达量较低;且在茎顶端初生生长阶段表达量较高,而在下部次生生长节间表达量较低,可能其主要参与巨桉的初生生长或者负调控次生生长;另外在盐胁迫、茉莉酸甲酯处理下其均被抑制表达;而在水杨酸、缺硼、缺磷处理下均上调表达。这说明EgrEXPA8和EgrEXPA10在巨桉响应逆境胁迫时起到重要作用,且呈现出相似的调控方式。     

RNA synthesis in aphids, Lachnus tropicalis

Aphid cells contain unique RNAs with molecular weights of 1.2 × 10⁶ and 0.6 × 10⁶. These RNAs are larger in amount in the nucleus and labeled rapidly when the insect is injected with [³H] uridine. Seemingly, the 28S and 18S rRNAs are completed at the cost of these RNAs.     

穗醋栗MYB10基因克隆结构分析及功能验证

为探究花色苷合成相关转录因子MYB10在不同颜色穗醋栗果实着色差异的分子机理,通过cDNA末端快速扩增技术(rapid amplification of cDNA ends, RACE)法从果实花青素含量有较大差异的黑穗醋栗(Ribes nigrum L.)、红穗醋栗(Ribes rubrum L.)和白穗醋栗(Ribes album L.)中分别克隆出MYB10基因,分别命名为RnMYB10 (KY786107)、RrMYB10 (KY786108)和RaMYB10(MW660848)。系统发育分析表明,RnMYB10和RrMYB10在进化上具有同源性。实时荧光定量PCR(real-time quantitative PCR,RT-qPCR)结果表明:黑穗醋栗各时期果实中MYB10表达量均高于红穗醋栗且远远高于白穗醋栗。随着果实直径加大颜色加深,RnMYB10和RrMYB10表达量呈现先上升后下降的趋势(在果实转色程度75%时达到最大值),RaMYB10表达量极低,几乎无表达。过表达RnMYB10和RrMYB10的拟南芥呈现紫色叶柄和叶片,过表达RaMYB10的拟南芥无明显变化。说明...     

Manipulation of aggressive behavior in adult DBA/2/Bg and C57BL/10/Bg male mice implanted with testosterone in silastic tubing

Adult DBA/2/Bg and C57BL/10/Bg male mice were orchidectomized and implanted subcutaneously with one of two doses of testosterone (T) in Silastic tubing. Steroid doses which resulted in significant atrophy and hypertrophy of accessory sex tissues were determined for each strain. Relative to sham-castrated vehicle-implanted controls, both hypophysiological and hyperphysiological implants caused augmentation of aggression in C57BL/10/Bg males. In DBA/2/Bg mice, the hypophysiological dose reduced and the hyperphysiological implant increased aggression scores. The results demonstrate that agonistic behavior in male mice is influenced by the titer of circulating T and emphasize the importance of genetic variance in the study of hormones and behavior.     

The immune response against Francisella tularensis live vaccine strain in Lpsn and Lpsd mice

Abstract The impact of Lps gene on the course of immune response against subcutaneous infection of mice with Francisella tularensis live vaccine strain was studied. Production and specificity of antibodies, cytotoxic responses of macrophages and NK-cells, spontaneous production ex vivo of cytokines IL-1α, IL-2, IL-4, IL-6, IL-10, IFN-γ, and TNF-α in spleen cell cultures in C3H/HeJ ( Lps ^d) mice in comparison with C3H/HeN ( Lps ^r) mice were tested. The value of LD₅₀ was significantly different in the two strains of mice (8.0 × 10³ cfu for C3H/HeJ versus 4.61 × 10⁵ cfu for C3H/HeN mice after subcutaneous inoculation). The production of NO₂ is also impaired in C3H/HeJ mice in the early intervals after infection. Thus, the defective Lps gene of C3H/HeJ mice influences both the level of innate resistance of mice to F. tularensis live vaccine strain infection and the process of induction and regulation of immune response against this intracellular bacterial pathogen.     

SAW1 is required for SDSA double-strand break repair in S. cerevisiae

SAW1, coding for Saw1, is required for single-strand annealing (SSA) DNA double-strand break (DSB) repair in Saccharomycescerevisiae. Saw1 physically associates with Rad1 and Rad52 and recruits the Rad1–Rad10 endonuclease. Herein we show by fluorescence microscopy that SAW1 is similarly required for recruitment of Rad10 to sites of Synthesis-Dependent Strand Annealing (SDSA) and associates with sites of SDSA repair in a manner temporally overlapped with Rad10. The magnitude of induction of colocalized Saw1-CFP/Rad10-YFP/DSB-RFP foci in SDSA is more dramatic in S and G2 phase cells than in M phase, consistent with the known mechanism of SDSA. We observed a substantial fraction of foci in which Rad10 was localized to the repair site without Saw1, but few DSB sites that contained Saw1 without Rad10. Together these data are consistent with a model in which Saw1 recruits Rad1–Rad10 to SDSA sites, possibly even binding as a protein–protein complex, but departs the repair site in advance of Rad1–Rad10.     

Extra components in kinetoplast DNA preparations from Crithidiafasciculata

Kinetoplast DNA from the order Kinetoplastidae (trypanosomatids) exists as large associations (molecular weight 4 × 10¹⁰), made up of about 104 small, probably circular, molecules, commonly known as ‘minicircles’. These minicircles were originally thought to be identical in base composition, suggesting that the coding capacity of kinetoplast DNA is very restricted. However, linear molecules have also been observed in preparations of kinetoplast DNA, which, if they contain unique sequences, could represent additional genetic information. This linear DNA has been assumed to be derived from the kinetoplast, but the possibility of it being nuclear contamination has not been definitely ruled out. Work presented in this paper demonstrates that nuclear DNA contamination may indeed be present in kinetoplast DNA prepared by a commonly used method.     

UNC-6/netrin and SLT-1/slit guidance cues orient axon outgrowth mediated by MIG-10/RIAM/lamellipodin

BACKGROUND: Axon migrations are guided by extracellular cues that can act as repellants or attractants. However, the logic underlying the manner through which attractive and repulsive responses are determined is unclear. Many extracellular guidance cues, and the cellular components that mediate their signals, have been implicated in both types of responses. RESULTS: Genetic analyses indicate that MIG-10/RIAM/lamellipodin, a cytoplasmic adaptor protein, functions downstream of the attractive guidance cue UNC-6/netrin and the repulsive guidance cue SLT-1/slit to direct the ventral migration of the AVM and PVM axons in C. elegans. Furthermore, overexpression of MIG-10 in the absence of UNC-6 and SLT-1 induces a multipolar phenotype with undirected outgrowths. Addition of either UNC-6 or SLT-1 causes the neurons to become monopolar. Moreover, the ability of UNC-6 or SLT-1 to direct the axon ventrally is enhanced by the MIG-10 overexpression. We also demonstrate that an interaction between MIG-10 and UNC-34, a protein that promotes actin-filament extension, is important in the response to guidance cues and that MIG-10 colocalizes with actin in cultured cells, where it can induce the formation of lamellipodia. CONCLUSIONS: We conclude that MIG-10 mediates the guidance of AVM and PVM axons in response to the extracellular UNC-6 and SLT-1 guidance cues. The attractive and repulsive guidance cues orient MIG-10-dependant axon outgrowth to cause a directional response.