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At least 487 records · Page 27Linked to original sources

PubMed2005

Combination of immunomagnetic separation with real-time PCR for rapid detection of Salmonella in milk, ground beef, and alfalfa sprouts.

A rapid, specific, and sensitive method for detecting Salmonella spp. in pasteurized milk, ground beef, and alfalfa sprouts was developed. The method combined immunomagnetic separation with a real-time PCR assay based on the double-stranded DNA binding dye SYBR Green I. The primers used produced a product with a melting temperature of 87+/-0.5 degrees C during the PCR assay by amplifying a 284-bp sequence from the invasive gene (invA) of Salmonella. The method was successful in detecting 20 Salmonella strains, but the expected PCR product was not formed by any of 11 other bacterial strains. To test this combined method for the monitoring of Salmonella, Salmonella enterica serotype Newport was inoculated into 52 samples each of pasteurized milk, ground beef, and alfalfa sprouts. Following a 10-h nonselective enrichment step in buffered peptone water, cells were removed by immunomagnetic separation and DNA extracted using the High Pure PCR template preparation kit. The DNA produced was used as a template in the real-time PCR assay. When spiked pasteurized milk, ground beef, and alfalfa sprout samples were analyzed by this protocol, an initial inoculum of 1 CFU/ml, 25 CFU/25 g, and 1.5 CFU/25 g, respectively, was detectable within 13 h. These results indicate that the combination of immunomagnetic separation and real-time PCR assay was a highly specific and sensitive method for the rapid detection of Salmonella.

Animals↗

PubMed2001

Auxin and heat shock activation of a novel member of the calmodulin like domain protein kinase gene family in cultured alfalfa cells.

A calmodulin like domain protein kinase (CPK) homologue was identified in alfalfa and termed MsCPK3. The full-length sequence of cDNA encoded a 535 amino acid polypeptide with a molecular weight of 60.2 kDa. The deduced amino acid sequence showed all the conserved motifs that define other members of this kinase family, such as serine-threonine kinase domain, a junction region and four potential Ca2+ -binding EF sites. The recombinant MsCPK3 protein purified from E. coli was activated by Ca2+ and inhibited by calmodulin antagonist (W-7) in in vitro phosphorylation assays. The expression of MsCPK3 gene increased in the early phase of the 2,4-D induced alfalfa somatic embryogenesis. Heat shock also activated this gene while kinetin, ABA and NaCl treatment did not result in MsCPK3 mRNA accumulation. The data presented suggest that the new alfalfa CPK differs in stress responses from the previously described homologues and in its potential involvement in hormone and stress-activated reprogramming of developmental pathways during somatic embryogenesis.

2,4-Dichlorophenoxyacetic Acid↗

PubMed2001

A comparison of recommended and reduced insecticide regimes in alfalfa.

The objective of this study was to evaluate the effects of the recommended rates of insecticides applied at two different population levels of the alfalfa weevil, Hypera variabilis Hbst. (Coleoptera: Curculionidae) and the reduced levels of insecticides on alfalfa weevil, predators and yield in 1998 and 1999. More specific objective was to search for options to reduce insecticide usage while implementing an effective pest control, thereby developing more economically and environmentally sound alfalfa production systems. The treatments used included: i) recommended rate insecticide (malathion) at population level of 5 weevils/sweep, ii) recommended rate at population level of 10 weevils/sweep, iii) 50% of recommended rate at population level of 10 weevils/sweep, iv) stripe spray of 50% rate at population level of 10 weevils/sweep, where insecticides were applied only on 0.5 m stripes in plots while the next 0.5 m stripes were not treated and v) control. The results indicated that the recommended rate insecticide application at 5 weevils/sweep level provided the highest reduction in the weevil densities in both years. No significant differences occurred among the weevil densities in the plots received the recommended rate, the 50% rate and the stripe spray insecticide treatments at 10 weevils/sweep level. Predators were in general more abundant in the insecticide treated plots compared to the control. The insecticide applications, except the stripe spray in 1999, led to increases in yields.

Animals↗

PubMed2004

Expression of UDP-glucose dehydrogenase reduces cell-wall polysaccharide concentration and increases xylose content in alfalfa stems.

The primary cell-wall matrix of most higher plants is composed of large amounts of uronic acids, primarily D-galacturonic acid residues in the backbone of pectic polysaccharides. Uridine diphosphate (UDP)-glucose dehydrogenase is a key enzyme in the biosynthesis of uronic acids. We produced transgenic alfalfa (Medicago sativa) plants expressing a soybean UDP-glucose dehydrogenase cDNA under the control of two promoters active in alfalfa vascular tissues. In initial greenhouse experiments, enzyme activity in transgenic lines was up to seven-fold greater than in nontransformed control plants; however, field-grown transgenic plants had only a maximum of 1.9-fold more activity than the control. Cell-wall polysaccharide content was lower and Klason lignin content was higher in transgenics compared to the nontransformed control. No significant increase in pectin or uronic acids in the polysaccharide fraction was observed in any line. Xylose increased 15% in most transgenic lines and mannose concentration decreased slightly in all lines. Because of the complexity of pectic polysaccharides and sugar biosynthesis, it may be necessary to manipulate multiple steps in carbohydrate metabolism to alter the pectin content of alfalfa.

Biomass↗

PubMed2004

[Instability of a cryptic plasmid in Sinorhizobium meliloti P108 during symbiosis of it with alfalfa Medicago sativa].

Instability of cryptic plasmids in Sinorhizobium meliloti laboratory strains SKhM1-188, DM7-R, and P108 as well as in their clones isolated from nodules of alfalfa grown during a long-term microvegetation experiment (120 days) was studied. The isolated clones of strains SKhM1-188 and DM7-R manifested stable inheritance of plasmids, whereas 12.7-14.0% of clones with changed plasmid profile were detected in a population of clones from strain P108. These segregants were designated as P108c. Segregants P108c exhibited significantly decreased symbiotic effectiveness, nitrogenase activity, and the competitiveness with respect to alfalfa, compared to the original strain P108. It was established that a 80-kb deletion occurred in a larger of two cryptic plasmids (240 and 230 kb) of segregants P108c. It was concluded that genetic rearrangements are possible in rhizobial clones that did not undergo structural transformation and retained viability in the nodule during the natural vegetation period of alfalfa.

Medicago sativa↗

PubMed2005

[Degradation process and plant diversity of alfalfa grassland in North Loess Plateau of China].

Vegetation recovery and reconstruction is the key of ecosystem restoration in the North Loess Plateau. With the Liudaogou catchment of Shenmu County, Shaanxi Province as test area, this paper studied the dynamics of the plant diversity of artificial alfalfa (Medicago sativa) grassland during its degradation process. The results showed that the degradation process of the grassland could be divided into three stages, i.e., artificial alfalfa grassland stage (1-6 yrs), transitional stage from artificial to natural grassland (6-10 yrs), and secondary natural grassland stage dominated by Stipa bungeana ( >10 yrs). In the whole degradation process from 1-to 30-yrs, 32 species belonging to 13 families and 28 genera were found, of which, 90% appeared in the initial 6-yrs. The dynamics of accumulatively appeared family, genus, and species in the vegetation succession process were well described by logarithmic function. During the process of community succession, the species richness (Gleason index and Margalef index),plant diversity (Shannon-Wiener index), and Pielou evenness index were changed with a similar tendency, i.e., increased more rapidly at the first stage, and the climax phase appeared in the second stage. The plant diversity and evenness decreased slightly, and gradually became stable later. Planting alfalfa could markedly accelerate the natural vegetation succession process in the forest-steppe ecotone of Loess Plateau, mainly due to the intensive soil water consumption of artificial grassland, which accelerated the process of soil aridification. To develop artificial grasslands in the Loess Plateau is an optimal joint between accelerating natural vegetation recovery and increasing farmers' income by stockbreeding, and is also a favorable paradigm both for the ecological and for the economic benefits in the eco-environmental construction in West China.

Biodiversity↗

PubMed2006

[Composition diversity of lactic acid bacteria (LAB) community Al2 used for alfalfa silage].

Alfalfa is the most important forage grass that is difficult to ensile for good quality. Using silage inoculants are the important way for preservation of alfalfa silage. Through continuous restricted subcultivation, a lactic acid bacteria (LAB) community Al2 was selected from well-fermented alfalfa silage. Plate isolation and Denaturing Gradient Gel Electrophoresis (DGGE), construction of 16S rDNA clone library were used to identify the composition diversity of Al2 community, with 7 strains detected, and they were all belonged to Lactobacillus. The composition ratios of the 7 strains were 55.21%, 19.79%, 14.58%, 3.13%, 3.13%, 3.13%, 1.03% according to 16S rDNA clone library. Al2-1i, Al2-2i, Al2-3i, corresponding to L. plantarum (99.9%), L. kimchii (99.4%), L. farciminis (100%) were detected by plate isolation. Among 3 isolates, Al2-1i had the highest ability of dropping pH and producing lactic acid, and the amount of lactic acid was reach to 18g/L at 24h cultivated in MRS media. The ability of dropping pH and producing lactic acid of Al2-3i was the lowest. From DGGE profiles, the dominant strains in Al2 community were L. plantarum and L. kimchii. L. plantarum was detected during the whole process, and L. kimchii was detected in the later phase.

Electrophoresis, Polyacrylamide Gel↗

PubMed1987

[Evaluation of different levels of amaranth flour (vegetative parts), in substitution for alfalfa flour for growing rabbits].

The vegetative part of the amaranth plant could be a useful resource for animal feeding due to its chemical characteristics and to the high yield obtained when harvested between 45 and 60 days after planting. Six diets were evaluated in order to determine the feed efficiency of amaranth (Amaranthus hypochondriacus) for growing rabbits. The diets contained dehydrated amaranth leaves and stalks in levels of 0, 15, 30, 45 and 60%, to replace equal amounts of alfalfa leaf meal. An additional diet containing 60% of steam-treated leaves and stalks of amaranth, for five minutes, prior to dehydration and milling, was also tested. The amaranth leaf meal contained 17.8% protein and 12.4% crude fiber as compared with the alfalfa leaf meal which contained 22.0% protein and 23.3% crude fiber. In terms of weight gain, feed intake, feed efficiency, carcass weight and serum proteins, the results indicated that amaranth leaf meal can efficiently replace alfalfa leaf meal up to 15% of the total weight of the diet. Higher levels, however, caused growth retardation and a pathological picture characterized by interstitial nephrosis and edema, more easily observed at a 60% level in the diet. Results also revealed that a steam treatment improves the nutritive quality of the amaranth meal.

Animal Feed↗

PubMed1995

Cloning novel alfalfa cyclin sequences--a RACE-PCR approach.

Cyclins are a complex group of proteins involved in regulation of the eukaryotic cell division cycle via their interaction with cyclin dependent kinases (Cdks). Cyclin gene sequences have been cloned from a number of plant species, including alfalfa, but the diversity of these genes suggests that there are many plant cyclins which have yet to be characterized. A RACE-PCR strategy has been adopted for cloning cyclin gene sequences expressed during direct somatic embryogenesis in alfalfa. RT-PCR with nested degenerate primers was used to amplify the highly conserved "cyclin box" region of a novel A-like cyclin mRNA sequence expressed after induction of somatic embryogenesis. The sequence of this PCR product was used to design primers for 5'- and 3'-RACE protocols. 5'-RACE using a modified SLIC (single strand ligation to single stranded cDNA) procedure revealed considerable sequence heterogeneity in the N-terminal region of the coding sequence with several closely related sequences apparent. Conventional 3'-RACE generated a single cyclin sequence. The complete coding sequence of one member of this A-like cyclin subgroup has been obtained by this RACE strategy and confirmed by PCR amplification and sequencing of alfalfa genomic DNA.

Amino Acid Sequence↗

PubMed1994

Evaluation of two supplements for the prevention of alfalfa bloat.

Poloxalene and a mineral mixture feed supplement patented for the treatment of emphysema, polyarthritis, and other pectin related diseases were tested in two trials for their ability to prevent bloat in cattle fed fresh alfalfa. Each trial had a crossover design using three Jersey steers with rumen fistulas per group. Each trial period continued until the total number of cases of bloat reached > or = 24. Treatments were given at 0800 each day. The mineral mixture was given at 100 g/d and 190 mg/kg body weight per day in the first and second trials, respectively. Poloxalene, which was tested only in the second trial, was given at 23 mg/kg body weight per day. Each group of steers was then fed 200 kg of freshly harvested alfalfa in the vegetative to early bloom stages of growth at 0830. In the first trial, only 69% as many cases of bloat occurred on the mineral mixture as on the control treatment, but no significant difference was detected in the second trial. The potency of the alfalfa may have been higher in the second trial, when forage dry matter was lower, magnesium and soluble nitrogen were higher, and bloat occasionally occurred twice a day. Bloat did not occur when the steers were treated with poloxalene. In these trials, poloxalene was completely effective in preventing bloat, but the mineral mixture was only partially so.

Animal Feed↗

PubMed1994

Toxic hepatopathy and photosensitization in cattle fed moldy alfalfa hay.

Cattle in 2 herds developed type-3 photosensitization after eating moldy alfalfa hay. Clinical signs included severe epidermal necrosis of unpigmented skin and marked decrease of milk production (herd 1). One herd had 18% mortality. Values for serum gamma-glutamyltransferase, lactate dehydrogenase, aspartate transaminase, and serum bilirubin were high in affected cows. Biliary epithelial degeneration and necrosis affecting the smaller bile ductules is the most consistent histologic lesion. Biliary hyperplasia, early portal fibroplasia, hepatocellular vacuolar degeneration and necrosis, and cholestasis were commonly seen. Mold growth on the alfalfa hay associated with prolonged wet weather prior to harvest was common to both herds. The cases reported here document hepatoxicosis and photosensitization associated with feeding moldy alfalfa hay grown in southeastern United States.

Alkaline Phosphatase↗

PubMedDate not supplied

[Nucleotide sequence of internal transcribed spacers and 5.8S rDNA for the ribosomal operon from alfalfa Medicago sativa and cotton Gossypium hirsutum L].

The 708- and 769-bp fragments from alfalfa and cotton containing the 3'-end of the 18S gene, the internal transcribed spacer. 1 (ITS1) the 5.8S gene, ITS2, and the 5'-end of the 28S gene were obtained using primers to the 18S and 28S ends of rDNA from tomato by a polymerase chain reaction. These sequences were cloned into pTZ19R. The 5.8S rDNA, ITS1 and ITS2 nucleotide sequences of alfalfa and cotton were determined. Comparative analysis of nucleotide sequences of alfalfa and cotton showed large hypervariability of spacer regions and conservatism of 5.8S rDNA sequence.

Base Sequence↗

PubMed1993

Comparative survival of first-stage larvae of small lungworms Muellerius capillaris and Neostrongylus linearis of goats in alfalfa and ryegrass plots.

Faeces from a goat infected with M capillaris and from another infected with N linearis were used to assess survival of first-stage larvae under natural conditions in northwest France in mid-autumn, at the high-risk period. The faeces were deposited on 3 plots covered respectively with ryegrass, short cut alfalfa and high alfalfa. The desiccation of faeces and survival of larvae were tested at 1, 7, 14 and 21-d intervals. Survival of M capillaris was much higher than that of N linearis in all plots. The lower desiccations were observed in the high alfalfa plot whereas the best survivals were found in the 2 other plots. Vegetation where faeces are deposited may thus play a part in the accumulation of parasitic risk on a pasture.

Analysis of Variance↗

PubMed1996

Developmental and cell cycle regulation of alfalfa nucMs1, a plant homolog of the yeast Nsr1 and mammalian nucleolin.

We report here the isolation and characterization of the nucMs1 alfalfa cDNA, whose predicted amino acid sequence structurally resembles the yeast Nsr1 protein and animal nucleolins. These proteins consist of an N-terminal acidic domain, centrally located RNA recognition motifs (RRMs), and a C-terminal glycine- and arginine-rich domain. In comparison with animal nucleolins that contain four RRMs, NucMs1 more closely resembles the yeast Nsr1 protein, which contains only two RRMs. A NucMs1 C-terminal peptide antibody specifically recognized a 95-kD nucleolar protein in alfalfa cells that changed its localization in a cell cycle-dependent manner. The nucMs1 transcript and p95nucMs1 protein levels correlated with cell proliferation, and nucMs1 gene expression was found to be induced in the G1 phase upon mitogenic stimulation of G0-arrested leaf cells. In situ hybridization analysis of different alfalfa organs during various developmental stages showed that nucMs1 gene expression is highest in root meristematic cells, but it is also found in other meristematic cells of the plant body. nucMs1 expression is tightly linked to cell proliferation but does not depend on a particular cell cycle phase. No nucMs1 expression was observed in cells that had exited the cell cycle and were undergoing differentiation or polar growth, indicating that nucMs1 may not be necessary for processes other than cell proliferation.

Animals↗

PubMed1996

Primary photosensitization related to ingestion of alfalfa silage by cattle.

A herd of 650 Holstein cows was examined for skin disease. Approximately 400 of the lactating adults were affected, but heifers, calves, and nonlactating cows were clinically normal. The condition was characteristic of primary photosensitization. Milk production of the affected cows was normal. Affected cows did not appear to be ill, and none of the cows was icteric. Three of 7 cows had high serum gamma-glutamyltransferase activities, but in the other 4 cows, activity was within the reference range. Serum activities of other hepatic enzymes were within reference ranges in the 7 cows that were examined. Hepatic biopsy specimens from 3 cows were normal. Specimens from 4 other cows had changes that ranged from minimal to mild, chronic, lymphoplasmacytic periportal hepatitis to acute, random, necrotizing hepatitis. Development of photosensitivity was related to ingestion of alfalfa silage. Acetone extracts of the alfalfa silage, but not of other feedstuffs, were found to inhibit growth of Candida albicans under ultraviolet light. Cows experimentally fed a diet composed exclusively of the alfalfa silage developed skin lesions after 6 days, but did not have detectable serum concentrations of phylloerythrin.

Animals↗

PubMed1998

Regulation of single-strand RNA synthesis of alfalfa mosaic virus in non-transgenic cowpea protoplasts by the viral coat protein.

We have compared the RNA synthesis of alfalfa mosaic virus in complete (by RNAs 1, 2 and 3) and incomplete infections (by RNAs 1 and 2) of cowpea protoplasts. Both viral RNA polymerase activity and accumulation of viral RNA were measured. By annealing RNA in solution with 32P-labelled probes of plus and minus polarity followed by treatment with ribonucleases, we determined viral RNAs quantitatively in both single- and double-stranded RNA fractions. The accumulation of single-stranded RNA of positive polarity differed considerably between the two types of infection (250 ng vs. less than 1 ng per 10(5) protoplasts), although viral RNA polymerase activities as measured in vitro and the concentrations of minus RNA were similar. Since the method also measured fragmented RNA, this difference is probably not due to lack of protection of viral RNA by coat protein during incomplete infection. Synthesis of single-stranded plus RNA requires either RNA 3 itself or one of its gene products. We postulate that coat protein is the stringent regulator of alfalfa mosaic virus genomic expression.

Alfalfa mosaic virus↗

PubMed1993

Coat protein stimulates replication complexes of alfalfa mosaic virus to produce virion RNAs in vitro.

Viral replication complexes (RCs) were gradient-purified from cowpea mesophyll protoplasts 21 h after inoculation with alfalfa mosaic virus. These membranous structures incorporate [32P]UMP into double- and single-stranded RNAs in the absence of added template. When coat protein is added prior to the reaction the incorporation in both RNA fractions is stimulated several times. Part of the single-stranded product RNAs are released from the RCs. The stimulation of incorporation in high molecular mass RNAs by coat protein can be mimicked only to a certain extent by addition of a ribonuclease inhibitor or of an excess of viral RNA prior to the reaction. This shows that the coat protein is not only protecting the product of the RCs against degradation by ribonuclease, but that it is stimulating the synthesis and release of viral RNAs from RCs as well. This leads to the hypothesis that with alfalfa mosaic virus some coat protein is necessary for the release of messenger RNA from the RC. The hypothesis explains why the viral genome RNAs, although they are of messenger polarity, cannot start a replication cycle in the absence of coat protein: RCs containing the parental RNAs could be formed but no amplification of them could take place since no messenger RNAs needed for the production of viral polymerase proteins would be released into the cytoplasm.

Alfalfa mosaic virus↗

PubMed2004

Cofolding organizes alfalfa mosaic virus RNA and coat protein for replication.

Alfalfa mosaic virus genomic RNAs are infectious only when the viral coat protein binds to the RNA 3' termini. The crystal structure of an alfalfa mosaic virus RNA-peptide complex reveals that conserved AUGC repeats and Pro-Thr-x-Arg-Ser-x-x-Tyr coat protein amino acids cofold upon interacting. Alternating AUGC residues have opposite orientation, and they base pair in different adjacent duplexes. Localized RNA backbone reversals stabilized by arginine-guanine interactions place the adenosines and guanines in reverse order in the duplex. The results suggest that a uniform, organized 3' conformation, similar to that found on viral RNAs with transfer RNA-like ends, may be essential for replication.

3' Untranslated Regions↗