Employment of stressful conditions during culture production to enhance subsequent cold- and acid-tolerance of bifidobacteria

AIMS: This study examined whether exposure of early stationary phase Bifidobacterium longum and B. lactis cells to various combinations of reduced temperature, reduced pH and starvation would enhance the cells' subsequent cold- and/or acid-tolerance. METHODS AND RESULTS: Survival of B. longum in growth medium at 6 degrees C significantly (P < 0.05) increased as a result of starving cells for 30 or 60 min without any simultaneous decrease in temperature or pH. Acid-tolerance of B. lactis (at pH 3.5 in synthetic gastric fluid) increased significantly when the growth medium pH was decreased from 6.0 to 5.2 and cells experienced 30 or 60 min of starvation. Enhanced B. lactis acid-tolerance persisted through 8-11 weeks of -80 degrees C storage in the pH 5.2 growth medium. Upon addition to milk during yogurt manufacture, these cells initially had enhanced acid-tolerance relative to untreated cells but untreated cells became equally acid-tolerant during the first 2.5 h of yogurt manufacture. CONCLUSIONS: The cold- and acid-tolerance of bifidobacteria vary widely, but may be significantly increased by application of sub-lethal stress to early stationary phase cells during culture production. SIGNIFICANCE AND IMPACT OF THE STUDY: The enhancement of B. lactis acid-tolerance observed in this study may be of potential importance in the production of effective ready-to-consume probiotic dietary supplements.     

Chloroplast ultrastructure in leaves of tobacco plants with the introduced gene for the acyl-lipid Δ9-desaturase from <Emphasis Type="Italic">Synechococcus vulcanus</Emphasis> at normal and low temperature

Ultrastructural changes in chloroplasts of tobacco plants (Nicotiana tabacum L.) with the introduced desC gene for the acyl-lipid Δ9-desaturase from the thermophilic cyanobacterium Synechococcus vulcanus were investigated during plant acclimation to cold. Control plants were transformed with an empty pGA482 binary vector. At optimum growth temperature, a decreased number of grana and thylakoids and an increased number of plastoglobules and their larger area were observed in transgenic plants when compared to control ones. In control plants, acclimation to cold (6 days at 10°C) resulted in the larger areas of chloroplasts and grana. These changes indicated starting cold-induced injuries manifested in swelling of the stroma and a slight decrease in the total number of thylakoids in the chloroplast. In contrast, transgenic plants responded to cold by reducing the chloroplast, granal, and plastoglobule areas. Meantime, the number of thylakoids per granum increased noticeably. The total number of thylakoids in the chloroplast increased from 123 to 203. It was concluded that expression of the acyl-lipid Δ9-desaturase gene in tobacco plants provided for the formation of the cell ultrastructure similar to one characteristic of cold-tolerant plants.     

转天山雪莲sikPIP3基因烟草的获得及抗逆性鉴定

利用PCR技术从实验室建立的天山雪莲DNA文库中克隆了天山雪莲质膜水孔蛋白基因sikPIP3,构建了植物表达载体pBI121-sikPIP3,通过农杆菌介导法转化烟草品种NC89,经PCR和RT-PCR检测证明目的基因成功导入并得到了表达,采用水分胁迫进行抗旱分析和采用冷冻胁迫进行抗寒性分析。结果显示:(1)克隆出具有水孔蛋白基因特性的sikPIP3基因。(2)经断水干旱处理,转基因烟草的生长表型优于野生型烟草,特别是在断水9d的情况下,野生型烟草已经完全萎蔫,转基因烟草萎蔫症状较轻;生理指标测量结果显示,转基因烟草的相对电导率和MDA的含量低于野生型烟草,相对含水量和CAT活性高于野生型烟草。试验表明转sikPIP3烟草的抗旱性高于野生型烟草。(3)经不同温度胁迫处理,转基因烟草的生长表型优于野生型烟草,特别是在-4℃冷处理6h情况下,野生型烟草已经完全萎蔫,转基因烟草只出现少量伤斑;生理指标分析结果表明,转基因烟草的相对电导率和MDA的含量低于野生型烟草,CAT活性高于野生型烟草。试验表明转sikPIP3烟草的抗寒性高于野生型烟草。综合结果表明:sikPIP3基因在抗逆基因工程方面具有较高的应用前景。     

柴达木盆地唐古特白刺果实调节免疫、抗疲劳和耐寒冷作用研究

采用环磷酰胺所致的免疫功能低下的模型小鼠试验柴达木盆地唐古特白刺果实的免疫调节作用,并进行了抗疲劳和耐寒冷试验。结果显示：唐古特白刺果实3．6g/kg灌胃剂量能显著增强模型小鼠单核巨噬细胞吞噬功能、迟发性超敏反应;1．8g/kg和3．6/kg灌胃剂量能显著提高模型小鼠血清溶血素生成能力;3．6g/kgg灌胃剂量可显著减轻环磷酰胺对小鼠胸腺DNA含量的抑制作用;3．6g/kg灌胃剂量能显著延长小鼠的游泳时间和爬杆时间,并能提高小鼠在低温的存活时间。实验表明唐古特白刺果实具有调节免疫、抗疲劳和耐寒冷作用。     

Growth light level and photon absorption by cells of Chlamydomonas rheinhardii,Dunaliella tertiolecta (Chlorophyceae,Volvocales), Scenedesmus obliquus (Chlorophyceae,Chlorococcales) and Euglena viridis (Euglenophyceae,Euglenales)

Reductions in the growth light level (40 to 6 μmol m^-2 s^-1) resulted in increases in chlorophyll and protein per cell for all of the species examined. Only Dunaliella tertiolecta exhibited a reduction in chlorophyll a:b ratio with decreases in the photon flux density. However, the specific absorption coefficient (ā? _i ) normalized to chlorphyll a (ā? _a remained invariant for all of the microalgae studied. Constant values for the specific absorption coefficient normalized to the total pigment content (ā? _a+b ) were also found for the species Chlamydomonas rheinhardii, Euglena viridis and Scenedesmus obliquus. In contrast ā? _a+b for D. tertiolecta decreased with a reduction in light level due to an increase in the proportion of chlorophyll b. Differences in ā? _i were related to cell size and pigment content and possible reasons for the constancy of ā? _a discussed. Increases in the absorption cross sections (¯sQ _a ) were also found at reduced light levels due to an increase in the absorptance per cell (α_cell). The lower α_cell for D. tertiolecta, compared with C. rheinhardii was exactly compensated for by a larger light-capturing area. Although the increase in α_cell does not compensate for the reduction in the incident light level, it does reduce this range by half on an absorbed light basis.     

江苏地区烟粉虱的越冬研究

通过室内模拟寒潮、越冬大棚内虫量观察,以及冬后田间调查,研究江苏地区烟粉虱的越冬状况。结果表明,在0℃以上的环境温度下,72 h内各虫态烟粉虱均不会造成急性致死,在-2℃的环境温度下,1 h内低龄若虫开始死亡,24 h后高龄若虫开始死亡,6 h后卵全部死亡。在-4℃的环境温度下,6 h后烟粉虱所有虫态均全部死亡。在单膜覆盖的大棚内,烟粉虱成虫在-2.5℃的寒潮下全部死亡,在双膜覆盖的大棚内,烟粉虱可以安全越冬。研究认为,在江苏地区,烟粉虱可以在日光温室或双膜覆盖的大棚内越冬,苏南部分地区可以在单膜覆盖的大棚内越冬。     

美洲黑杨与大青杨杂种无性系耐寒性的初步研究

以美洲黑杨与大青杨杂种无性系1年生扦插苗为材料,测定其低温胁迫中多个生理指标的变化。结果表明,6个杂种无性系SOD、POD和APX活性在低温胁迫中均表现出先上升后下降的趋势,而电解质渗漏率、活性氧含量、CAT和GR活性保持逐渐升高的趋势。无性系间差异显著,其中DU146和DU147电解质渗漏率和活性氧含量最低,而5种抗氧化酶活性均最高,表明其具有较强的耐寒性;而DU135和DU150电解质渗漏率和活性氧含量最高,而5种抗氧化酶活性均最低,表明其耐寒性差。     

Rapid cold-hardening response in Ophraella communa LeSage (Coleoptera: Chrysomelidae), a biological control agent of Ambrosia artemisiifolia L.

In China, an unintentionally introduced beetle Ophraella communa shows good control efficiency against invasive common ragweed, Ambrosia artemisiifolia. With the aim of understanding the capacity of O. communa to overcome sub-zero low temperature rapidly, the longevity, survival and fecundity of the beetle adults were studied at conditions of different rapid cold-hardening in the laboratory. The results showed that approximately 20% female or male individuals still survive at ?12°C for 2 h, thus the discriminating temperature for rapid cold-hardening of both female and male beetles is ?12°C. The survival, longevity and fecundity of adult beetles were significantly affected by rapid cold-hardening treatments. Compared to the control, the survival rates of adult beetles experiencing 2 h of rapid cold-hardening at 2, 5 and 8°C prior to exposure to its discriminating temperature (?12°C) were significantly increased. Adult longevity and fecundity were significantly higher after pre-treatment at 2, 5 and 8°C for 2 h than those of the control. Both longest longevity and maximum fecundity of adult beetles were observed at the rapid cold-hardening of 2°C. Our experiment implies that the cold-tolerance ability of O. communa can be rapidly increased after rapid cold-hardening treatments, which reveals an important ecological significance for population establishment and expansion of O. communa in the invaded areas of common ragweed.     

不同核桃品种的耐寒性及其渗透调节机制

以‘香玲’、‘鲁W06-1’、‘西洛3号’3个核桃品种（优系）为材料,对低温胁迫各材料的耐受情况及相应渗透调节物质含量和总蛋白质表达的变化情况进行分析,以揭示低温胁迫下不同核桃品种（优系）的渗透调节机制和蛋白质组分差异。结果显示:（1）3个核桃品种（优系）休眠枝条解剖结构分析表明,‘鲁W06-1’一年生休眠枝条的木质部比率最大,‘西洛3号’的韧皮部厚度显著大于其他两个品种（优系）。（2）随胁迫温度降低和胁迫时间的延长,枝条相对电导率（REC）逐渐升高,且‘西洛3号’的相对电导率均大于‘鲁W06-1’和‘香玲’。（3）核桃枝条可溶性糖、可溶性蛋白和游离脯氨酸含量均随温度降低先增加后减小,但各品种(优系)增至最大值的温度不同,其中‘鲁W06-1’枝条积累可溶性蛋白的速度和幅度较大,且各低温胁迫处理后的枝条可溶性糖和游离脯氨酸的含量也更高;3种渗透调节物质含量之间均呈极显著正相关关系,可溶性糖与游离脯氨酸含量的相关系数达0.844,表明二者对低温胁迫的应答反应密切相关。（4）在不同胁迫温度和胁迫时间处理后,核桃枝条的蛋白质表达谱带总体相似,但检测出6条表达量明显增加的差异条带,它们的分子量范围为38.9～87.9 kD。研究表明,3个核桃品种（优系）的耐寒性强弱为‘鲁W06-1’>‘香玲’>‘西洛3号’;低温胁迫下,核桃枝条迅速积累可溶性蛋白,然后积累可溶性糖和游离脯氨酸,并以耐寒性较强品种（优系）积累速度更快、积累量更大,且耐寒性较强的品种（优系）枝条蛋白谱带出现较多表达量增加的条带。