海藻糖酶

The results showed that the mutant contained high trehalose content and high cell survival rate kept even after 4 h 18 %vol alcohol treatment (no respiration deficiency existed), which indicated that high trehalose content could protect cells membrane and prevent DNA loss of mitochondria and exosmosis of intracellular materials.

结果表明，中性海藻糖酶缺失突变株细胞海藻糖含量较高，用18 %vol的酒精处理4 h后仍可保持高的细胞存活率，并且无呼吸缺陷型的出现，说明高海藻糖含量可以保护细胞膜，防止线粒体DNA丢失和胞内物质的外渗，与酵母的酒精耐性之间存在一定的关系。

First,through monofactorial experiment,we studied the influences of pH,temperature and substrate concentration on acidic trehalase activity of baker's yeast.

首先通过单因素实验研究了pH值、温度、底物浓度对面包酵母酸性海藻糖酶活性的影响，然后通过正交试验得出了酸性海藻糖酶活性测定的适宜条件：海藻糖浓度1.6%、温度30℃、pH 4.0、酵母菌泥接种量为0.02g/mL。

To test the influence of trehalose accumulation on response to freeze stress, we constructed all single and double gene disruption mutants of the trehalase genes ATH1, NTH1 and NTH2 and examined the changes of their trehalose accumulations and their freeze tolerance.

为了检测冷冻压力下海藻糖积累的作用，我们构建了海藻糖酶基因ATH1、NTH1和NTH2的各种单基因突变株和双基因突变株，并对突变株的海藻糖积累情况和耐冷冻能力进行了检测。

The activity of neutral trehalase lost and trehalose concentration has been increased three times after knocking out neutral trehalase gene in S. cerevisiae.

将酿酒酵母细胞内中性海藻糖酶基因敲除后，中性海藻糖酶活性丧失，海藻糖含量提高3 倍。

First, through monofactorial experiment, we studied the influences of pH, temperature and substrate concentration on acidic trehalase activity of baker's yeast. Then we designed an orthogonal experiment to detective the optimal condition of ascertaining acidic trehalase activity. The optimal conditions are: substrate concentration 1.6%, pH 4.0, temperature 30 and seed volume 0.02g/mL compressed yeast.

首先通过单因素实验研究了pH值、温度、底物浓度对面包酵母酸性海藻糖酶活性的影响，然后通过正交试验得出了酸性海藻糖酶活性测定的适宜条件：海藻糖浓度1.6%、温度30℃、pH 4.0、酵母菌泥接种量为0.02g/mL。

To determine the role of neutral trehalase during storage stability of spores, we constructed a M.anisopliae RNAi transformant and an overexpression transformant. Analysis of these transformants revealed that the expression level of the neutral trehalase gene in RNAi transformant(No 688) was reduced to 50% of which found in wild type and the overexpression transformant(No 113) showed approximatlely 2-fold higher levels of neutral trehalase mRNA than that found in wild-type control.

构建了中性海藻糖酶基因RNA干扰载体和超表达载体，采用电转法和基因枪法转化金龟子绿僵菌CQMa102，获得了RNA干扰转化子和超表达转化子，比较这两种转化子和出发菌株的mRNA表达情况，发现RNA干扰转化子中性海藻糖酶mRNA表达受到抑制，而超表达转化子中性海藻糖酶mRNA的表达量是出发菌株的2倍。

Sugar and protein content in the haemolymph after fed with transgenic rice Total sugar content in the haemolymph of SSB fed with KMD1 was significantly lower than that of the control, while trehalase activity was significantly higher than that of the control, but there were no significant differences on trehalose concentration and glucose concentration between the treatment and the control. However, there were no significant differences on all the four components mentioned above between SSB fed on MSB and the control.

四、取食转基因抗虫水稻后二化螟幼虫血淋巴糖代谢和蛋白质浓度的变化五龄二化螟幼虫取食KMD1后血淋巴总糖含量显著低于取食秀水11对照的，海藻糖酶活性显著高于对照，而海藻糖含量和葡萄糖含量与对照组相比都无显著差异；取食MSB后二化螟血淋巴总糖含量、海藻糖含量、葡萄糖含量以及海藻糖酶活性与取食明恢86的对照相比，均差异不显著。

The glucose-6-phosphate dehydrogenase increased during overw intering,indicating that reduced condition of cell might keep the cell in pr oper function even in cold temperature,and also for cryoprotectant synthesis.The changes in trehalase activity showed that low activity was for keeping higher tre halose content and high activity for removing it in the spring.

越冬期间，体内葡萄糖-6-磷酸脱氢酶活性增高所产生的还原型烟酰胺腺嘌呤二核苷酸磷酸，可为细胞在亚低温状态下发挥正常功能以及体内抗冻保护性物质的合成提供还原动力，同时通过调节体内海藻糖酶活性来维持越冬期间较高含量的海藻糖和移除春季体内累积的过多的海藻糖。

Analysis of these mutants revealed that the expression level of the neutral trehalase gene in RNAi mutant (Ma688 strain) was reduced to 42% of which found in wild-type strain and the over-expression mutant (Ma113 strain) showed approximatlely 2-fold higher levels of neutral trehalase mRNA than that found in wild-type control. Then the transformants Ma688 and Ma113 were chosen for closer examination, including trehalose content, storage longevity , thermotol

利用Northern blot 分析这几个突变菌株中性海藻糖酶基因mRNA 的含量，选择中性海藻糖酶基因超表达2 倍和中性海藻糖酶基因被抑制到42%的两个转化菌：Ma113 和Ma688，将其与出发菌株进行分生孢子内海藻糖含量的测定，孢子储藏

The results showed that the conidia of the RNAi transformant accumulated more trehalose, lost viability more slowly during prolonged storage and showed reduced heat shock sensitivity than the wild type, and the overexpression transformant has reverse results. These results indicate that neutral trehalase in M.anisopliae control trehalose concentration via degradation of trehalose, and indirectly influnce the viability of conidia during prolonged storage and the resistance to heat stress.

以上研究结果表明，金龟子绿僵菌CQMa102孢子体内海藻糖是由中性海藻糖酶降解的，海藻糖含量与耐储性和耐热性具有正相关性，中性海藻糖酶在虫生真菌金龟子绿僵菌孢子中的生理功能是通过降解分子内海藻糖调控海藻糖含量，从而影响其对不良环境的抗性，如储藏性、耐热性。

As she looked at Warrington's manly face, and dark, melancholy eyes, she had settled in her mind that he must have been the victim of an unhappy attachment.

每逢看到沃林顿那刚毅的脸，那乌黑、忧郁的眼睛，她便会相信，他一定作过不幸的爱情的受害者。

Maybe they'll disappear into a pothole.

也许他们将在壶穴里消失