Starvation induces physiological changes that act on the cryotolerance of Lactobacillus acidophilus RD758

The relationship between lactose starvation and cryotolerance was investigated in Lactobacillus acidophilus RD758. Cryotolerance was measured from the acidification activity of cells recovered after 18-h lactose starvation. It was compared to that of nonstarved cells, both of them in a stationary phase and in the same medium. This measurement allowed quantifying the initial acidification activity before freezing, as well as the loss of acidification activity during freezing and the rate of loss during frozen storage. Even if initial acidification activity was similar for nonstarved and starved bacteria, the latter displayed a significantly better resistance to freezing and frozen storage at -20°C. To investigate the mechanisms that triggered these cryotolerance phenomena, the membrane fatty acid composition was determined by gas chromatography, and the proteome was established by 2-D electrophoresis, for starved and nonstarved cells. The main outcome was that the improved cryotolerance of starved cells was ascribed to two types of physiological responses as a result of starvation. The first one corresponded to an increased synthesis of unsaturated, cyclic, and branched fatty acids, to the detriment of saturated fatty acids, thus corresponding to enhanced membrane fluidity. The second response concerned the upregulation of proteins involved in carbohydrate and energy metabolisms and in pH homeostasis, allowing the cells to be better prepared for counteracting the stress they encountered during subsequent cold stress. These two phenomena led to a cross-protection phenomenon, which allowed better cryotolerance of Lb. acidophilus RD758, following cellular adaptation by starvation.     

棉花在广西南宁的越冬耐冷性鉴定和遗传分析

在广西南宁对不同种质来源的棉花材料及其杂交后代进行越冬耐冷性鉴定,为多年生棉花耐冷育种提供基础材料。在开展最佳处理时间和处理温度筛选的基础上,采用5个耐冷生理生化指标(相对电导率、可溶性糖、可溶性蛋白、丙二醛和脯氨酸)和露地越冬指标综合鉴定63份材料的越冬耐冷性,并运用系统聚类的方法将其聚为4大类,即耐冷型23份、中度耐冷型19份、中度冷敏型9份和冷敏型12份,分别占供试材料的36.51%、30.16%、14.29%和19.05%。所有耐冷型材料均和多年生海岛棉种质有关,说明多年生海岛棉在越冬耐冷性育种上具有很高的利用价值。供试棉花材料越冬性的细胞质效应不显著,杂交后代的越冬性有向亲本的平均类型回归的趋势。     

Lactobacillus,Bifidobacterium and Lactococcus response to environmental stress: Mechanisms and application of cross-protection to improve resistance against freeze-drying

The review deals with lactic acid bacteria in characterizing the stress adaptation with cross-protection effects, mainly associated with Lactobacillus, Bifidobacterium and Lactococcus. It focuses on adaptation and cross-protection in Lactobacillus, Bifidobacterium and Lactococcus, including heat shocking, cold stress, acid stress, osmotic stress, starvation effect, etc. Web of Science, Google Scholar, Science Direct, and PubMed databases were used for the systematic search of literature up to the year 2020. The literature suggests that a lower survival rate during freeze-drying is linked to environmental stress. Protective pretreatment under various mild stresses can be applied to lactic acid bacteria which may enhance resistance in a strain-dependent manner. We investigate the mechanism of damage and adaptation under various stresses including heat, cold, acidic, osmotic, starvation, oxidative and bile stress. Adaptive mechanisms include synthesis of stress-induced proteins, adjusting the composition of cell membrane fatty acids, accumulating compatible substances, etc. Next, we reveal the cross-protective effect of specific stress on the other environmental stresses. Freeze-drying is discussed from three perspectives including the regulation of membrane, accumulation of compatible solutes and the production of chaperones and stress-responsive proteases. The resistance of lactic acid bacteria against technological stress can be enhanced via cross-protection, which improves industrial efficiency concerning the survival of probiotics. However, the adaptive responses and cross-protection are strain-dependent and should be optimized case by case.     

Heat shock and cold shock in Deinococcus radiodurans

On the basis of acquired thermotolerance and cryotolerance, the optimal heat shock and cold shock temperatures have been determined for Deinococcus radiodurans. A heat shock at 42°C maximized survival at the lethal temperature of 52°C and a cold shock at 20°C maximized survival after repeated freeze-thawing. Enhanced survival from heat shock was found to be strongly dependent on growth stage, with its greatest effect shortly after phase. Increased synthesis of a total of 67 proteins during heat shock and 42 proteins during cold shock were observed by two-dimensional polyacrylamide gel electrophoresis (2D PAGE) and autoradiography. Eight of the most highly induced heat shock proteins shown by 2D PAGE were identified by MALDI-MS as Hsp20, GroEL, DnaK, SodA, Csp, Protease I and two proteins of unknown function.     

Effect of culture period and cell density on regrowth following cryopreservation of embryogenic suspension cultures of Norway Spruce and Sitka spruce

Changes in cryotolerance, sedimented cell volume and dry weight were determined during a 21-day culture period for embryogenic suspension cultures of Norway spruce (Picea abies) and Sitka spruce (Picea sitchensis). Maximum cryotolerance was obtained for both species when the cultures were harvested in the phase of stationary growth in terms of dry weight. For P. abies, the culture period that resulted in maximum cryotolerance was similar to the culture period that resulted in maximum formation of mature embryos after 10 weeks of maturation. The initial cell density of the P. abies cultures is an important factor in affecting regrowth after cryopreservation and it was found that adjustment of the sedimented cell volume to 50% (v/v) resulted in maximum regrowth. This revised version was published online in June 2006 with corrections to the Cover Date.     

Acid adaptation of Lactobacillus delbrueckii subsp. bulgaricus induces physiological responses at membrane and cytosolic levels that improves cryotolerance

Aims: This work aimed at clarifying the physiological responses of Lactobacillus delbrueckii subsp. bulgaricus CFL1 cells after exposure to acidification at the end of fermentation, in relation to their cryotolerance. Methods and Results: Cells acidified at the end of the fermentation (pH 5·25 for 30 min) had their cryotolerance improved as compared to the reference condition (pH 6·0). By analyzing the cytosolic proteome, it was established that changes occurred in the synthesis of 21 proteins, involved in energy metabolism, nucleotide and protein synthesis and stress response. Acidification also induced a slight decrease in unsaturated to saturated and cyclic to saturated membrane fatty acid ratios. Conclusions: Lactobacillus bulgaricus CFL1 was able to develop a combined physiological response at both membrane and cytosolic levels. This acid adaptation was referred as a cross-protection phenomenon as it allowed the cells to become more tolerant to cold stress. Significance and Impact of the Study: This study increased knowledge concerning the physiological mechanisms that explained the cross-protection by acid adaptation. It may be useful for improving cryotolerance of lactic acid bacteria, either in cells banks or in an industrial context.