Molecular characterization of a stable antisense chalcone synthase phenotype in strawberry (Fragaria x ananassa)

An octaploid (Fragaria x ananassa cv. Calypso) genotype of strawberry was transformed with an antisense chalcone synthase (CHS) gene construct using a ripening related CHS cDNA from Fragaria x ananassa cv. Elsanta under the control of the constitutive CaMV 35S promoter via Agrobacterium tumefaciens. Out of 25 transgenic lines, nine lines showed a reduction in CHS mRNA accumulation of more than 50% as compared to the untransformed cv. Calypso control. The antisense CHS construct was found to be integrated into the genome, with a copy number ranging from one to four. The pigmentation of the fruit was only affected when less than 5% of the control CHS expression level was detected. A stable antisense phenotype over a period of 4 years was obtained in the primary transgenic lines at a rate of 1:20. As a consequence of the reduced activity of CHS, the levels of anthocyanins, flavonols, and proanthocyanidins were downregulated and precursors of the flavonoid pathway were shunted to the phenylpropanoid pathway leading to highly increased levels of cinnamoyl glucose (520% of control), caffeoyl glucose (816% of control), and feruloyl glucose (1092% of control) as well as p-coumaryl alcohol (363% of control) and p-coumaryl-1-acetate (1079% of control), which occur only as trace components in untransformed control fruits. These results demonstrate that the introduction of an antisense CHS construct in strawberry results in an unpredictable biochemical phenotype, thereby confirming that CHS function is an important regulatory point of substrate flow between the flavonoid and the phenylpropanoid pathways.     

Venograms for Use in Laminitis Treatment

Venography is a very valuable tool that can be used in the diagnosis and treatment of laminitis. It can be used to assess vascular damage in the hoof before changes in radiographic detail occur. When used at onset, the venogram proves to be most valuable. They can not only give baseline measurements but can also show the level of damage that has already occurred. Initial venograms allow farriers and veterinarians to evaluate what degree of corrective treatment is necessary for the best outcome. In addition, a series of venograms can show the process of pathology as well as recovery. From here, treatment can be altered to accommodate the case.     

Antioxidants in raspberry: on-line analysis links antioxidant activity to a diversity of individual metabolites

The presence of antioxidant compounds can be considered as a quality parameter for edible fruit. In this paper, we studied the antioxidant compounds in raspberry (Rubus idaeus) fruits by high-performance liquid chromatography (HPLC) coupled to an on-line postcolumn antioxidant detection system. Both developmental and genetic factors were assessed by comparing fruits from a single cultivar of different ripening stages and by comparing ripe fruits of 14 raspberry cultivars, respectively. The HPLC-separated antioxidant compounds were identified using HPLC-photodiode array coupled to mass spectrometry (quadrupole time-of-flight tandem mass spectrometry), using a reference lock mass for determining accurate masses. The dominant antioxidants could be classified as anthocyanins, ellagitannins, and proanthocyanidin-like tannins. During fruit ripening, some anthocyanins were newly produced, while others, like cyanidin-3-glucoside, were already present early in fruit development. The level of tannins, both ellagitannins and proanthocyanidin-like tannins, was reduced strongly during fruit ripening. Among the 14 cultivars, major differences (>20-fold) were observed in the levels of pelagonidin type anthocyanins and some proanthocyanidin type tannins. The content of ellagitannins varied approximately 3-fold. The findings presented here suggest that the content of individual health-promoting compounds varies significantly in raspberry, due to both developmental and genetic factors. This information will assist in the future development and identification of raspberry lines with enhanced health-promoting properties.     

外源茉莉酸对大豆中异戊烯黄酮Glyceollins及其前体Glycinol累积的影响

以中黄13号大豆种子为材料,取生长8天的健康子叶,在其表面挖出约2×2mm的伤口,将50μl不同浓度的茉莉酸溶液和1mM的硝酸银溶液滴在伤口处,黑暗中(25℃)保温不同时间后取样,经甲醇提取,用高压液相色谱(HPLC)测定其中异戊烯化异黄酮glyceollins及其前体glycinol 的累积情况.结果显示,健康子叶中没有glyceollins和glycinol的累积,1mM硝酸银比茉莉酸具有更强的诱导glyceollins和glycinol累积的能力,glyceollins的累积与茉莉酸浓度呈负相关.在各种处理中,glyceollinⅠ、Ⅱ、Ⅲ所占比例分别为:50%～80%、10%～40%和10%～20%,但是glyceollin三种异构体的比例随保温时间的延长而有所变化,glyceollin Ⅰ比例逐渐下降,glyceollin Ⅱ所占比例逐渐升高,在1mM硝酸银处理的子叶中表现更为明显.     

Stress-induced biosynthesis of dicaffeoylquinic acids in globe artichoke

Leaf extracts from globe artichoke ( Cynara cardunculus L. var. scolymus) have been widely used in medicine as hepatoprotectant and choleretic agents. Globe artichoke leaves represent a natural source of phenolic acids with dicaffeoylquinic acids, such as cynarin (1,3-dicaffeoylquinic acid), along with its biosynthetic precursor chlorogenic acid (5-caffeoylquinic acid) as the most abundant molecules. This paper reports the development of an experimental system to induce caffeoylquinic acids. This system may serve to study the regulation of the biosynthesis of (poly)phenolic compounds in globe artichoke and the genetic basis of this metabolic regulation. By means of HPLC-PDA and accurate mass LC-QTOF MS and MS/MS analyses, the major phenolic compounds in globe artichoke leaves were identified: four isomers of dicaffeoylquinic acid, three isomers of caffeoylquinic acid, and the flavone luteolin 7-glucoside. Next, plant material was identified in which the concentration of phenolic compounds was comparable in the absence of particular treatments, with the aim to use this material to test the effect of stress application on the regulation of biosynthesis of caffeoylquinic acids. Using this material, the effect of UV-C, methyl jasmonate, and salicylic acid treatments on (poly)phenolic compounds was tested in different globe artichoke genotypes. UV-C exposure consistently increased the levels of dicaffeoylquinic acids in all genotypes, whereas the effect on compounds from the same biosynthetic pathway, for example, chlorogenic acid and luteolin-7-glucoside, was much less pronounced and was not statistically significant. No effect of methyl jasmonate or salicylic acid was found. Time-response experiments indicated that the level of dicaffeoylquinic acids reached a maximum at 24 h after UV radiation. On the basis of these results a role of dicaffeoylquinic acids in UV protection in globe artichoke is hypothesized.     

Hoof capsule distortion: understanding the mechanisms as a basis for rational management.

Understanding the basic mechanisms of hoof capsule distortion enables the clinician to unravel the sometimes complex mix of abnormalities, identify early warning signs, and manage existing problems more effectively. Distortion of the hoof capsule can often seem like a complicated puzzle by the time the veterinarian is called to examine the lame horse. Evaluating each of the components of the hoof capsule, both individually and as an integrated unit, allows the clinician to tease apart the tangle of primary and secondary problems and come up with effective options for countering or attenuating the forces responsible for the distortion. Using high-quality radiography and developing an eye for detail regarding the physical and radiographic characteristics of the foot allow the clinician to identify subtle abnormalities, thereby averting a clinical problem or addressing the problem while it is still reversible. With existing problems, developing these skills enhances the clinician's ability to interpret the degree of damage accurately and to devise mechanical solutions that create an environment in which healing and restoration of function are maximized.     

Radiographic imaging of the equine foot.

Radiographic examination of the equine foot can provide the veterinarian and farrier with a wealth of information. Positioning and selection of exposure factors are of central importance if one is to produce radiographs of maximum diagnostic value.