谷氨酰胺的抗疲劳生化机制研究

用两批各 48只小鼠分 4组 (n =12 )每天分别灌喂生理盐水和 3 4mmol·kg-1的谷氨酰胺、谷氨酸和天冬酰胺 ,连续 10d。结果显示 ,三者都可提高小鼠血清、肝和骨骼肌中的肌酸磷酸激酶活性 ,降低乳酸脱氢酶活性 ,并显著增加肝、骨骼肌糖原积累。其中有相同碳架结构的谷氨酰胺、谷氨酸能极显著提高小鼠的游泳耐力 ,但与谷氨酰胺有相等酰胺氮的天冬酰胺作用不强。结果提示 ,谷氨酰胺提高机体抗疲劳作用与其碳架结构有关     

Effect of idazoxan on permeability of inflammatory blood-brain barrier model in vitro

AIM: To study the effect of idazoxan on the permeability of inflammatory blood-brain barrier (BBB) model in vitro and the expression of tight junction protein ZO-1.METHODS: In vitro BBB model was established by murine brain endothelial cell line bEnd.3 incubated for 7 d. The cells were treated with TNF-α (10 nmol/L) for additional 24 h to establish the inflammatory BBB model, which was pretreated with IDA at doses of 50, 100 and 200 μmol/L, respectively. The permeability was measured using fluorescein isothiocyanate-conjugated dextran (FD-40, MW 40,000), the expression of ZO-1 was detected by Western blot analysis, the distribution of ZO-1 was observed by immunofluorescence, and the mRNA expression of MMP-9/TIMP-1 was measured by RT-PCR.RESULTS: After incubated for 7 d, b.End3 cells converged to be confluent monolayer with low permeability. The inflammatory BBB model induced by TNF-α treatment displayed much higher permeability with decreased expression of tight junction protein ZO-1, destroyed distribution of ZO-1 and increased mRNA expression of MMP-9. When pretreated with IDA, the permeability was greatly decreased, the expression of ZO-1 was greatly increased, the abnormal distribution of ZO-1 was greatly ameliorated and the mRNA expression of MMP-9 was obviously reduced. The effect was most significant in IDA (200 μmol/L)-pretreated group (P<0.01).CONCLUSION: IDA directly acts on brain endothelial cells to reduce the expression of MMP-9, increase the expression of tight junction protein ZO-1 and ameliorate the destroyed distribution of ZO-1 in the inflammatory BBB, thus reversing the abnormally elevated permeability in a inflammatory BBB model in vitro induced by TNF-α.     

Introduction of new traits into cotton through genetic engineering: insect resistance as example

The main goal of gene transfer into cotton is the development of insect-resistant varieties. The stakes are important since cotton protection against insects uses almost 24% of the world's chemical insecticides market, which is not without consequences on the environment. The first approach was to introduce and express in the cotton genome, genes from the bacterium Bacillus thuringiensis (B.t.) which produces entomopathogenic toxins. The development of an efficient Agrobacterium tumefaciens mediated transformation system was the first step. The expression of B.t. genes was studied and synthetic genes more adapted to a plant genome have been constructed. Studies on their expression in cotton is underway. The second focus was to develop strategies that would minimize the risks of inducing insect resistance. The main approach is to associate several genes coding for entomopathogenic proteins with different modes of action. Genes encoding protease inhibitors were chosen. One possibility is to associate a B.t. gene and a gene encoding a protease inhibitor. Several protease inhibitors were tested in artificial diets on major pests of cotton. The corresponding genes have been introduced into the cotton genome. These various orientations of the research program will be presented. This revised version was published online in July 2006 with corrections to the Cover Date.     

转甜菜碱醛脱氢酶基因马铃薯的抗旱耐盐性

通过根癌农杆菌介导法将甜菜碱醛脱氢酶(BADH)基因导入马铃薯栽培品种甘农薯2号, 经PCR、Southern杂交和Northern杂交证明BADH基因已整合到马铃薯基因组中并在转基因植株中转录和表达。测定表明对照植株没有BADH酶活性, 各转化株系在胁迫前后BADH酶活性近似, 在2~11 U之间。BADH酶活性与叶片的相对电导率呈一定的负相关(y= –3.7738x+57.083, r=0.989^**)。在NaCl和PEG胁迫下, 转基因植株生长正常, 株高比对照提高0.41~1.00 cm, 单株重量比对照增加10%~35%, 说明外源BADH基因的导入提高了马铃薯植株对干旱和盐碱的抗性。     

转甜菜碱醛脱氢酶基因水稻种子萌发和苗期耐盐性的研究

用0.0、3.0、5.0g/L NaCl对2个转甜菜碱醛脱氢酶(BADH)基因水稻51-22、52-7与受体亲本中花8进行萌发期盐胁迫试验.结果表明,盐胁迫下转基因材料受抑程度小于受体亲本,尤以52-7表现明显.用0.0、3.0、5.0g/L NaCl对51-22、52-7和中花8在幼苗期进行14 d盐胁迫试验.结果表明,低盐胁迫时幼苗受盐害较小,高盐胁迫时转基因材料耐盐性优于受体亲本;在盐胁迫7 d后,转基因材料体内脯氨酸含量明显高于受体亲本,同一品种随盐度增大,脯氨酸含量升高.     

野油菜黄单胞菌6-磷酸葡萄糖脱氢酶基因的初步分析

野油菜黄单胞菌野油菜变种是发酵工业生产黄原胶的菌种,对该菌8004菌株全基因组序列进行分析,发现基因组中有两个编码6-磷酸葡萄糖脱氢酶的基因,编号分别为XC1977和XC4082,同源性分析显示这两个基因演绎的氨基酸序列只有36.3%的相似性。为了解这两个基因的功能,采用自杀质粒pK18 mob对XC1977和XC4082分别进行诱变,构建这两个基因的非极性突变体,对突变体表型初步分析,发现XC1977和XC4082分别突变后不影响细胞的生长繁殖,但对胞外多糖产量的影响则不相同,XC1977突变使胞外多糖产量降低56.3%,而XC4082突变基本不影响胞外多糖产量,表明8004菌株的两个zwf基因在细胞中的功能不同,XC1977与细胞的胞外多糖合成产量有关。     

Hypoglycemic effect of lupin seed γ-conglutin in experimental animals and healthy human subjects

A lupin seed γ-conglutin-enriched preparation was tested in a glucose overload trial with both murine models and adult healthy volunteers. The results with rats showed a dose-dependent significant decrease of blood glucose concentration, which confirmed previous findings obtained with the purified protein. Moreover, three test-product doses equivalent to 630, 315, and 157.5 mg γ-conglutin, orally administered 30 min before the carbohydrate supply, showed a relevant hypoglycemic effect in human trials. Insulin concentrations were not significantly affected. The general hematic parameters did not change at all.This is the first report on the glucose-lowering effect of lupin γ-conglutin in human subjects.     

Effective nitrification inhibitors may improve fertilizer recovery in irrigated cotton

N fertilizer is often poorly recovered in irrigated cotton production, due to N loss through denitrification. We researched the ability of inhibitors to delay nitrition and reduce the availability of NO₃ ^- to denitrifying microorganisms and thus improve N fertilizer recovery, 2-Ethynylpyridine, etridiazole, and nitrapyrin proved highly effective nitrification inhibitors, although nitrification was evident several weeks after their application. CaC₂ was relatively ineffective, even when wax-coated to prolong the evolution of C₂H₂. Phenylacetylene and ethynylcyclohexanol were also ineffective, despite having a chemical structure similar to 2-ethynylpyridine. A strong association was identified between each compound's ability to inhibit nitrification and its capacity to improve N fertilizer recovery. In one experiment, N fertilizer recovery was increased by 50% with 2-ethynylpyridine, etridiazole, or nitrapyrin application, from 33% without inhibitors. The inhibitors had little effect on fertilizer recovery where N losses were relatively small. 3-Methyl pyrazole significantly increased N uptake and lint yield, but the nitrification inhibitors had no significant effect on N uptake or on yield in two of the three of the cotton crops. A laboratory study confirmed that nitrification inhibitor effectiveness declined in the order 2-ethynylpyridine>etridiazole>nitrapyrin>3-methyl pyrazole>phenylacetylene>CaC₂>ethynylcyclohexanolThis research was conducted at Australian Cotton Research Institute, CSIRO Division of Plant Industry, Locked Bag 59, Narrabri, NSW 2390, Australia     

Soil microbial functional diversity response following nematocide and biocide amendments in a desert ecosystem

The aim of the current study was to gain a better understanding of the changes that occur in soil microbial community and in its functional diversity as a result of the use of nematocide and biocide inhibitors in natural ecosystems. Both inhibitors are known to have a great effect on the nematode community and total biota, playing an important role in soil food web and biota interactions. The experiment was set up in the Negev Desert using sixteen 1×1 m soil plots, to which two chemical inhibitors were applied: (a) a biocide, to eliminate the whole biotic community; and (b) a nematocide, to eliminate the nematode community in soil. In addition, water treatment was applied to the same soil plots, while untreated soil plots were used as control. Microbial functional diversity, together with abiotic parameters such as soil moisture and total organic carbon, was tested monthly in soil samples collected from the 0-10 and 10-20 cm soil layers. The results of the abiotic parameters showed similar patterns in the two soil layers regardless of the inhibitor treatments. An increase in soil water content followed rainfall patterns. Total organic carbon was low during the wet season and increased during the dry seasons. The Shannon-Weaver index value for microbial functional diversity was found to increase in spring after the wet season in both soil layers. In the upper soil layer, an increase was observed both in the inhibitor and water treatments. However, the increase in the water treatment lasted longer compared to the increase observed in the inhibitor-treated soil plots. In the 10-20 cm soil layer, a different pattern was observed: an increase in microbial functional diversity was observed in the inhibitor-treated soil plots, while an increase in the water-treated soil plots was seen at a later stage. Principal Component Analysis was also conducted, revealing different patterns between inhibitors and water treatments on both a temporal scale, when changes from a homogeneous to heterogeneous consumption pattern were observed, and in the nature of communities that proliferate in the soil. Differences were also observed in the microbial community between the upper 0-10 and the lower 10-20 cm soil layers, where an opposite pattern of substrate consumption was observed. This study emphasizes the important role the biotic component plays in the soil of an arid climate, studying the long-term effects of key species elimination on the microbial community in desert soil.     

Measurement of coupled nitrification-denitrification in paddy fields affected by Terrazole,a nitrification inhibitor

Coupled nitrification-denitrification and potential denitrification were measured as ¹⁵N₂O and ¹⁵N₂ evolution rates in ammonium sulphate-treated rice soils with or without Terrazole [5-ethoxy-3 (trichloromethyl) 1,2,3 Thiadizole] under laboratory and field conditions. The greatest coupled nitrification-denitrification activity was found after drying and rewetting the soil, with maximum values of 322 ng N cm^–2 h^–1 in the laboratory and 90.8 ng N cm^–2 h^–1 in the field. These ¹⁵N₂O + ¹⁵N₂ evolution rates were about 10 times lower than potential denitrification in these soils. These results and the observed decrease in ¹⁵N₂O + ¹⁵N₂ evolution rate in soils treated with Terrazole (60% under laboratory conditions and 52% under field conditions) indicate that denitrification was limited by coupled nitrification-denitrification activity. Oxygen and previous addition of ammonium sulphate appear to control the rate of ¹⁵N₂O + ¹⁵N₂ evolution in ammonium sulphate-fertilised soils.