Callus Induction of Alfalfa and Evaluation of Callus Tolerance to Acid and Aluminum

Medicago is an important grass in the legume family,which is suitable to be grown in neutral or alkalescent soils,and hence,can be widely spread in south of China with low pH value.It is one of major objectives to cultivate acid-aluminum tolerant species.In the research,6 high-quality species with high-yielding potential were taken as materials to make evaluation on acid-aluminum tolerance by cell-culture technique,and the results showed that tolerance capacity from high to low was GT13R＞Muge 701=Muge 702＞Acrora＞AC-3＞Sheshou No.2=Medicago sativa.

Comparative Analysis of Six Triticum turgidum L. Subspecies for Acid and Aluminum Tolerance

The aluminum （A1） tolerance of hexaploid wheat and rye has been extensively studied. However, the A1 tolerance of tetraploid wheat （Triticum turgidum L.）, the AABB genome donor of hexaploid wheat, has not been well characterized. To understand the A1 tolerance of tetraploid wheat and identify potentially new sources of tolerance to acidic soils, 254 Triticum turgidum wheat lines, including six subspecies of T. turdigum ssp. dieoccoides, ssp. dicoccon, ssp. carthlicum, ssp. turgidum, ssp. durum, and ssp. polinicum, were screened for acid and A1 tolerance by measuring the root regenerate lengths and the tolerance indices in acid nutrient culture solution containing A1. Significant differences were observed either among or within tetraploid wheat subspecies on the RRL （root regenerate lengths）. The orders for average RRL among subspecies in pH 7.00, pH 4.50 and pH 4.50 ＋ 50μmol L^-1 A1^3＋ were carthlicum ≥ turgidum ≥ durum ≥ dicoccoides polinicum ≥ dicoccon, carthlicum ≥ turgidum ≥ durum ≥ polinicum ≥ dicoccon ≥ dicoccoides and durum ≥ dicoccoides carthlicum ≥ turgidum ≥polinicum ≥ dicoccon, respectively. Significant difference was also observed on the root acid tolerance index （RT11） and root A1 tolerance index （RTI2） either among or within tetraploid wheat subspecies. Compared to RTI2, RTI1 varied in a wide range but with large value. The orders of the average RTI1 and RTI2 among subspecies were polinicum ≥ dicoccon ≥ carthlicum ≥ dicoccoides ≥ turgidum ≥durum, and dicoccoides ≥ durum ≥ turgidum ≥polinicum carthlicum ≥dicoccon, respectively. Significant difference on acid and A1 tolerance was existed either among or within subspecies. Some potentially tolerance germplasms different from those derived from hexaploid wheat, including 42 acid tolerance and 41 A1 tolerance lines, were identified in 254 tetraploid wheat lines. They provide new sources for hexaploid wheat acid and A1 tolerance improvement.

Study on Acid Tolerance in Rhizobium meliloti and Its Symbiosis System

Soil pH is a main factor that affects the nodule formation and nitrogen fixation. The largely reduced nitrogen fixation effect caused by acid soil is the ma- jor limitation of alfalfa growth in the south of China. Thus selection of anti-acid nodule for inoculating on alfalfa is of great significance in practical production. Using nine candidate rhizobia （ Rhizobium meliloti L. ） as study objects, their colony diameters and the nodule number of each strain inoculated alfalfa variety were investigated at different pH conditions. The YNCY006 strain presented the strongest resistance to acid, while YNCY007 strain presented the weakest. For nedulation, GT13R alfalfa inoculated with YNCY006 or YNCY008 strains was the best symbiosis system in the acidic soil.

Effect of Lactose on Acid Tolerance of Yogurt Culture Bacteria

Lactose, the sugar naturally present in milk, provides energy to lactic acid bacteria used in fermented dairy foods. Increasing concentrations of lactose may improve survivability of lactic acid bacteria in the dairy foods and in human gut enhancing their probiotic benefits. Acid tolerance is an important probiotic characteristic. The objective was to determine the influence of lactose on acid tolerance of yogurt starter culture Streptococcus thermophiles ST-M5 and Lactobacillus bulgaricus LB-12. The M 17 broth was used for Streptococcus thermophiles ST-M5 and MRS broth was used for Lactobacillus bulgaricus LB-12. Lactose was added to both broths at 0% (control), 1%, 3%, and 5% (wt/vol). Both broths were acidified to pH 2.0. Upon sterilizing and tempering, both broths were inoculated. Acid tolerance was determined as viable counts in acidified broths after 120 minutes of incubations. In an incubation period of 2 hours, dilutions were plated every 30 minutes. Three replications were conducted. The highest acid tolerance for Streptococcus thermophiles ST-M5 and Lactobacillus bulgaricus LB-12, was observed in lactose concentration of 3% and 5% (wt/vol).

Divergent responses of growth rate and antioxidative system of ten Bacillus strains to acid stresses

Response of growth rate and antioxidative system of ten Bacillus strains to acid stresses was assayed.Strong acid treatment significantly decreased the growth rate of the strains.Acid stresses increased the GPX activity and GSSG content of the tested strains.Divergent changes occurred in ROS and antioxidative system(SOD,CAT,GR,MDA and GSH).Environmental changes including soil acidification exert obvi-ous stresses on soil ecosystems and influence soil microor-ganisms.In this study,ten microbial strains were incubated under different acid treatments to investigate responses of microbial growth and antioxidative system to acid stress.All the strains belong to Bacillus genus,but exhibit distinct ecological functions.We observed that these microbial strains had obviously different pH tolerance threshold,in spite of the close phylogenetic classification among strains.Acid stresses exerted significant effects on microbial antiox-idative system,including superoxide dismutase(SOD),cata-lase(CAT)and glutathione transferring enzymes(GPX and GR)and reactants(GSH and GSSH),but the effects were strain specific.Furthermore,we found acid stress effects on total variances of the investigated microbial antioxidative system along the first two principal components(PCs).Activities of CAT and SOD contributed substantially to PC1 that reflected obvious acid effects on NC7 and ZC4,and closely related to intracellular malondialdehyde content.The GSSG activities and GSH/GSSG contributed greatly to PC2 that unveiled acid stress effects on most of the microbial strains.Our results highlight substantially heterogeneous responses of microbial strains to acid stress and support that phylogenetic closeness does not imply functional similarity of soil microorganisms under environmental changes.

Influences of trans-trans farnesol,a membrane-targeting sesquiterpenoid,on Streptococcus mutans physiology and survival within mixed-species oral biofilms

Trans-trans farnesol （tt-farnesol） is a bioactive sesquiterpene alcohol commonly found in propolis （a beehive product） and citrus fruits, which disrupts the ability of Streptococcus mutans （S. mutans） to form virulent biofilms. In this study, we investigated whether tt-farnesol affects cell-membrane function, acid production and/or acid tolerance by planktonic cells and biofilms of S. mutans UA159. Furthermore, the influence of the agent on S. mutans gene expression and ability to form biofilms in the presence of other oral bacteria （Streptococcus oralis （S. oralis） 35037 and Actinomyces naeslundii （.4. naeslundil） 12104） was also examined. In general, tt-farnesol （1 mmol-L-1） significantly increased the membrane proton permeability and reduced glycolytie activity of S. mutans in the planktonic state and in biofilms （P〈0.05）. Moreover, topical applications of 1 mmol-L＂l tt-farnesol twice daily （1 min exposure/treatment） reduced biomass accumulation and prevented ecological shifts towards S. mutans dominance within mixed-species biofilms after introduction of 1% sucrose. S. oralis （a non-cariogenie organism） became the major species after treatments with tt-farnesol, whereas vehicle-treated biofilms contained mostly S. mutans （〉90% of total bacterial population）. However, the agent did not affect significantly the expression of S. mutans genes involved in acidogenicity, acid tolerance or polysaccharide synthesis in the treated biofilms. Our data indicate that tt-farnesoi may affect the competi- tiveness of S. mutans in a mixed-species environment by primarily disrupting the membrane function and physiology of this bacterium. This naturally occurring terpenoid could be a potentially useful adjunctive agent to the current anti-biofilm/anti-caries chemotherapeutic strategies.

Acid tolerance of lactate-utilizing bacteria of the order Bacteroidales contributes to prevention of ruminal acidosis in goats adapted to a high-concentrate diet

The rapid accumulation of organic acids,particularly lactate,has been suggested as the main cause of ruminal acidosis(RA)for ruminants fed high-concentrate diets.Previous research has shown that a gradual shift from low-to high-concentrate diets within 4 to 5 weeks effectively reduces the risk for RA.However,the mechanisms remain unknown.In this study,20 goats were randomly allocated into four groups(n=5)and fed with a diet containing a weekly increasing concentrate portion of 20%,40%,60%,and 80%over 28 d.At d 7,14,21,and 28,one group(named C20,C40,C60,and C80 according to the last concentrate level that they received)was killed and the ruminal microbiome was collected.Ruminal acidosis was not detected in any of the goats during the experiment.Nonetheless,ruminal pH dropped sharply from 6.2 to 5.7(P<0.05)when dietary concentrate increased from 40%to 60%.A combined metagenome and metatranscriptome sequencing approach identified that this was linked to a sharp decrease in the abundance and expression of genes encoding nicotinamide adenine dinucleotide(NAD)-dependent lactate dehydrogenase(nLDH),catalyzing the enzymatic conversion of pyruvate to lactate(P<0.01),whereas the expression of two genes encoding NAD-independent lactate dehydrogenase(iLDH),catalyzing lactate oxidation to pyruvate,showed no significant concomitant change.Abundance and expression alterations for nLDH-and iLDH-encoding genes were attributable to bacteria from Clostridiales and Bacteroidales,respectively.By analyzing the gene profiles of 9 metagenome bins(MAG)with nLDH-encoding genes and 5 MAG with iLDH-encoding genes,we identified primary and secondary active transporters as being the major types of sugar transporter for lactate-producing bacteria(LPB)and lactate-utilizing bacteria(LUB),respectively.Furthermore,more adenosine triphosphate was required for the phosphorylation of sugars to initiate their catabolic pathways in LPB compared to LUB.Thus,the low dependence of sugar transport systems and catabolic pathways on primary energy sources supports the acid tolerance of LUB from Bacteroidales.It favors ruminal lactate utilization during the adaptation of goats to a high-concentrate diet.This finding has valuable implications for the development of measures to prevent RA.

Effect of lit-moxa stimulation of Guanyuan(CV 4) acupoint on levels of lactic acid and super-oxide dismutase in skeletal muscle of rats

OBJECTIVE:To observe the effect of lit-moxa stimulating acupoint Guanyuan(CV 4) on lactic acid and super-oxide dismutase(SOD) in skeletal muscle after exercise exhaustion.METHODS:Twenty-eight adult male Sprague-Dawley rats were randomly divided into normal control group,exhausted control group,exercise group and moxibustion group using exercise training and mild heating with lit-moxa stick as treatment methods.The exhausted control group,moxibustion group and exercise group received an exhaustive swimming after 20 days of intervention.Swimming exhausted times were recorded.Lactic acid and SOD concentration in soleus muscle were detected and compared between every two groups.RESULTS:The swimming exhausted times of the moxibustion group and the exercise group were significantly increased compare to the exhausted control group(P < 0.05).The lactic acid of the exhausted control group was significantly increased comparing with the normal control group(P <0.05),and the lactic acid of the moxibustion group and the exercise group were significantly lower than that of the exhausted control group(P < 0.05).The SOD level of the exhausted control group was significantly decreased comparing with the normal control group(P < 0.05),and the SOD level of the moxibustion group and the exercise group were significantly higher than that of the exhausted control group(both,P < 0.05).There was no statistical difference between the moxibustion group and the exercise group.CONCLUSION:With lit moxa stick,heat stimulating acupoint of Guanyuan(CV 4) decreased the levels of lactic acid and SOD in rat's skeletal muscle.