Identification of QTL Affecting Protein and Amino Acid Contents in Rice

The phenotypes of protein and amino acid contents were measured in an F9 recombinant inbred line population derived from a cross between Zhenshan 97B and Delong 208. A total of 48 and 64 QTLs were identified in 2004 and 2005, respectively. The contribution of each QTL to the phenotypic variation ranged from 4.0% to 43.7%. Most QTLs co-localized, forming 29 QTL clusters on the chromosomes with three major ones detected in both years, which were mapped on chromosomes 1, 7 and 9, respectively. The two QTL clusters for amino acid content, qAa1 and qAa7, influenced almost all the traits with the allele from Zhenshan 97B, and the third QTL cluster for amino acid content, qAa9, increased the lysine content with the allele from Delong 208. A wide coincidence was found between the QTL detected under this study and the loci involved in amino acid metabolism pathways in nitrogen assimilation and transport, or protein biosynthesis. The results would facilitate the identification of candidate genes and could be used in marker-assisted selection for the favorable allele in rice quality improvement.

Effects of High-intensity Pulsed Electric Field on Amino Acid Content in Large-leaf Pu'er Tea Variety

[Objective] This study aimed to test the effects of high-intensity pulsed electric field on the amino acid content in Pu’er tea leaves. [Method] The amino acid contents of fermented and unfermented the leaves of large-leaf Pu’er tea variety produced in Yunnan Mengku, were measured before and after the treatment with high-intensity pulsed electric field （HPEF）. [Result] The difference of amino acid contents was notable between fermented and unfermented tea leaves produced in the same year. The amino acid content of unfermented tea leaves was larger than that in fermented tea leaves. The amino acid content in tea leaves decreased over time during storage. HPEF treatment under al the conditions improved the amino acid content in tea leaves. [Conclusion] This experiment provides theoretical foundation for the application of HPEF in processing of Pu’er tea leaves.

Effects of salt and rice flour concentration on microbial diversity and the quality of sour meat,a Chinese traditional meat

This study investigated the effects of salt(3%and 6%,m/m)and rice flour(10%and 20%,m/m)addition in sour meat,a traditional Chinese fermented meat.It was found that salt has greater effect than rice flour addition in spontaneous fermentation.Low-salt groups had lower pH and higher titratable total acid.In the low-salt groups,the dominant genera were Lactobacillus and Lactococcus,whereas Staphylococcus,Weissella,and Tetragenococcus were dominant in the high-salt groups.Higher total free amino acids and essential amino acids,organic acids,hexanoic acid ethyl ester and octanoic acid ethyl ester were found in the low-salt groups.The RDA analysis revealed that Lactococcus was closely related to product quality,with the S3F10(3%salt and 10%rice f lour)group outperforming the others in the sensory evaluation.Therefore,3%salt and 10%rice flour were considered more appropriate for the production of healthy and tasty fermented sour meats.

Physiological Processes Contributing to the Difference in Grain Amino Acid Content Between Two Hybrid Rice(Oryza sativa L.)Cultivars

Improving grain amino acid content of rice(Oryza sativa L.)is essential for the health of consumers.This study was conducted to identify the physiological processes that contribute to the higher grain amino acid content in hybrid rice cultivar Lingliangyou 268 compared to Luliangyou 996.The results showed that total amino acid content in grains was 9%higher in Lingliangyou 268 than in Luliangyou 996.There was no significant difference in grain nitrogen(N)content between Lingliangyou 268 and Luliangyou 996,while ratio of amino acid to N was 6%higher in Lingliangyou 268 compared to Luliangyou 996.A total of 16 differentially expressed proteins related to amino acid metabolism(e.g.,erythronate-4-phosphate dehydrogenase domain containing protein)were identified in grains between Lingliangyou 268 and Luliangyou 996.The identified proteins were involved in 10 molecular functions.Six of the 10 defined functions were related to binding(heterocyclic compound binding,nucleoside phosphate binding,nucleotide binding,organic cyclic compound binding,protein binding,and small molecule binding)and the other 4 defined functions were catalytic activity,enzyme regulator activity,hydrolase activity,and transferase activity.These results indicate that the higher grain amino acid content in Lingliangyou 268 compared to Luliangyou 996 is attributed to increased efficiency of converting N to amino acid that results from altered expression of proteins related to amino acid metabolism.

Determination and Analysis of Amino Acid Content in Maca from Different Habitats and Varieties

[Objectives]To determine and analyze amino acid content in Maca from different habitats and varieties.[Methods]Maca from two habitats and three colors were selected and a kind of turnip commonly used was to fake Maca.After optimizing the hydrolysis conditions,100 mg Maca samples were hydrolyzed in the concentration of 6 mol/L hydrochloric acid at 110℃for 24 h.The content range of amino acids in each sample was detected by HPLC.[Results]The average total content of amino acids in Maca was 9.52 g/100 g.Yunnan yellow Maca contained 16 kinds of amino acids,including 7 kinds of human essential amino acids,accounting for 17.2%of the total amino acids while Yunnan purple Maca held 16 kinds of amino acids,counting 7 kinds of human essential amino acids,accounting for 16.5%of the total amino acids.The amino acid ratio coefficient method also showed that the two kinds of Maca had higher nutritional value than the others.15 kinds of amino acids were detected in Yunnan black Maca,including 6 kinds of essential amino acids,accounting for 9.60%of the total amino acids.Among amino acids in Yunnan Maca,the highest content was Arg.A total of 12 kinds of amino acids,together with 6 essential human amino acids,were determined in Peru black Maca,accounting for 11.6%of the total amino acids.In Peru black Maca the highest concentration of amino acid was Pro.It was noteworthy that the amino acid species were comprehensive in Maca,and high level of amino acids resulted in high nutritional value.Its specific amino acid composition ratio was obviously different from Turnip,the counterfeit goods to deceive customers.[Conclusions]The method of automatic amino acid analyzer is simple and accurate,and can be used for the identification and content determination of amino acids in Maca as amino acid is an essential nutrient in Maca.

Effect of nitric oxide synthase inhibitor N^G-nitro-L-arginine on the content of amino acid in ischemic brain tissues of rats

BACKGROUND： Nitric oxide synthase （NOS） inhibrtors have been widely used to investigate the role of NO on cerebral ischemic injury, but the results are controversial. Moreover, it has been considered to aggravate the ischemic neuronal damage with the release of excessively excitatory amino acids （EAA） during cerebral ischemia. On the other hand, some inhibitory amino acid is suggested to be important for the neuronal protection against ischemic brain damage. Our study has recently showed that treatment with the NOS inhibitor NG-nitro-L-arginine （L-NA） reduced focal cerebral ischemic damage. The effect of L-NA on the contents of excitatory and inhibitory amino acid in the rat brain following cerebral ischemia is still unclear. OBJECTIVE： By evaluating the effect of NOS inhibitor, L-NA on the contents of aspartate, glutamate, glycine and γ-aminobutyric acid （GABA） in striatum, hippocampus and cortex in the rat brain following cerebral ischemia respectively, to investigate the beneficial effect of L-NA on cerebral ischemic injury and the possible mechanism. DESIGN： A randomized and controlled experiment SETTING ： Department of Pharmacology, Hebei Academy of Medical Sciences MATERIALS： A total of 42 male healthy SD rats （grade Ⅱ, weighting 250-300 g） were provided by the Experimental Animal Center of Hebei Province （Certification： 04036）. Aspartate, glutamate, glycine, GABA, L-NA and 2,3,5-triphenyltetrazolium chloride （TTC） were obtained from Sigma Chemicals Co, St Louis, MO, USA. HPLC-ultraviolet detector system consisted of Agilent 1100 HPLC. METHODS： The experiment was carried out in Department of Pharmrcology, Hebei Academy of Medical Sciences from June 2005 to June 2006. Rats were randomly divided into three groups： sham-operated group （n = 6）, ischemic group （n = 18）, L-NA group （n = 18）. The model of focal cerebral ischemia in rat was prepared with intraluminal line occlusion methods. In sham-operated rats, the external carotid artery was surgically prepared, but the filament was not inserted. Each group was further divided into 3 subgroups （n = 6 for each）： drugs were administrated at 2, 6 and 12 hours after the middle cerebral artery occlusion （MCAO） respectively. L-NA （20 mg/kg, ip） was administrated, twice a day, for 3 consecutive days. Same volume of normal saline was administrated in ischemic and sham operation groups. The changes of infarcted volume and the contents of amino acids were respectively assayed. Image analysis software was used for the measurement of the infarcted area. The results were expressed as a percentage of the infarcted volume of cerebral/volume of whole brain （IV%） in order to control for edema formation. The contents of aspartate, glutamate, glycine and GABA in striatum, hippocampus and cortex in the rat brain following cerebral ischemia were respectively measured by HPLC method. All data were analyzed with one-way ANOVA and Dunnett＇s test. MAIN OUTCOME MEASURES： （1） The volume of cerebral infarction; （2） The contents of aspartate, glutamate glycine and GABA in brain tissue after cerebral ischemia. RESULTS ： All 42 rats were involved in the final analysis. （1） Infarcted volume： Volume was 0 in sham-operated group. When L-NA was administrated at 2 and 6 hours after MCAO, the infarcted volume was （20.13±3.59）% and （23.12±5.84）% in L-NA group, which was not similar to that in ischemic group [（22.10±3.98）%, （25.38± 5.37）%, P〉 0.05]. However, the infarcted volume was markedly decreased compared with that of ischemic group when L-NA was administrated at 12 hours after MCAO [（26.11±3.55）% and （37.15±3.58）%, P 〈 0.01]. Changes of amino acid content： At 2 and 6 hours after ischemia, the contents of aspartate, glutamate, glycine and GABA in striatum, hippocampus and cortex in ischemic group were significantly increased compared with those in sham-operated group （ P〈 0.05-0.01）. However, contents in L-NA group were similar to those in ischemic group （P 〉 0.05）. At 12 hours after ischemia, the contents of aspartate [（0.21 ±0.06）, （0.36±0.05）, （0.29±0.12） mg/g] and glutamate [（0.55±0.06）, （0.78±0.10）, （0.52±0.10） mg/g] in striatum, hippocampus and cortex in L-NA group were significantly decreased compared with those in ischemic group [（0.49±0.17）, （0.63± 0.03）, （0.51±0.15） mg/g; （0.98±0.30）, （1.15±0.15）, （0.93±0.15） mg/g, P〈 0.05-0.01]. Glycine in hippocampus was （0.40±0.07） mg/g, which was higher than that in ischemic group [（0.21±0.07） mg/g, P 〈 0.05]. GABA in striatum, hippocampus and cortex was （0.93±0.10）, （0.62±0.12） and （0.81 ±0.10） mg/g, respectively, which was higher than that in ischemic group [（0.60±0.08）, （0.37±0.17）, （0.59±0.10） mg/g, P 〈 0.05-0.01]. CONCLUSION ： It may be concluded that L-NA have beneficial effect on ischemic cerebral injury in ischemic later stage in rats. The possible mechanism is that L-NA can decrease the contents of aspartate and glutamate, increase the contents of glycine and GABA.

Association Analysis of the Amino Acid Contents in Rice

The main objective of the present study was to identify simple sequence repeat （SSR） markers associated with the amino acid content of rice （Oryza sativa L.）. SSR markers were selected by prescreening for the relationship to amino acid content. Eighty-four rice landrace accessions from Korea were evaluated for 16 kinds of amino acids in brown rice and ganotyped with 25 SSR markers. Analysis of population structure revealed four subgroups in the population. Linkage disequilibrium （LD） patterns and distributions are of fundamental importance for genome-wide mapping associations. The mean r^2 value for all intrachromosomal loci pairs was 0.033. LD between linked markers decreased with distance. Marker-trait associations were investigated using the unified mixed-model approach, considering both population structure （Q） and kinship （K）. A total of 42 marker-trait associations with amino acids （P 〈 0.05） were identified using 15 different SSR markers covering three chromosomes and explaining more than 40% of the total variation. These results suggest that association analysis in rice is a viable alternative to quantitative trait loci mapping and should help rice breeders develop strategies for improving rice quality.

Chemical Constituents of Pedicularis longiflora var. tubiformis (Orobanchaceae), a Common Hemiparasitic Medicinal Herb from the Qinghai Lake Basin, China

Pedicularis longiflora var.tubiformis(Orobanchaceae)is an abundant parasitic herb mainly found in the Xiaopohu wetland of the Qinghai Lake Basin in Northwestern China.The species has an important local medicinal value,and in this study,we evaluated the chemical profile of its stems,leaves and seeds using mass spectrometry.Dried samples of stems,leaves and seeds were grinded,weighted,and used for a series of extractions with an ultrasonic device at room temperature.The chemical profiles for each tissue were determined using Gas Chromatography-Mass Spectrometry(GC-MS)and Liquid Chromatography-Mass Spectrometry(LC-MS).Twenty-seven amino acids and organic acids were identified and quantified from stems,leaves and seeds.The content of amino acids detected in leaves and seeds was higher than the amount found in stems.Six fla-vonoids were also detected,including isoorientin,orientin,luteolin-7-O-glucoside,luteolin,apigenin and tricin.The concentrations of luteolin-7-O-glucoside,luteolin and tricin were the highest and more concentrated in leaves,while that of orientin was the lowest and mainly found in stems.Soluble monosaccharides and oligosaccharides below tetramer were also examined,and our analyses detected the presence of arabitol,fructose,galacturonic acid,glucose,glucuronic acid,inositol,sucrose,and trehalose.This is the first study to identify and quantify the main components of amino acids,organic acids,flavonoids and soluble sugars from stems,leaves and seeds of P.longiflora var.tubiformis.Eight of the amino acids detected are essential for humans,highlighting the medicinal importance of this species.Results shown here can be used as a reference case to develop future studies on the chemical constituents of Pedicularis herbs and other medicinal plants from the Tibetan region.

Phytophthora Root and Crown Rot on Fruit Trees in Bulgaria

Phytophthora root and crown rot was found on the fruit trees in Bulgaria for the first time in the period 1998-1999. Monitoring of the disease spread from 2000 to 2007 points out incidence between 2 and 14 per cent, in some orchards and nurseries in the Southern part of Bulgaria. The following Phytophthora species were identified based on morphological and cultural characteristics, and temperature requirements： Phytophthora cactorum, Phytophthora citrophthora, Phytophthora drechsleri, Phytophthora cryptogea, Phytophthora hybrid specie and Pythium. Prevailing specie was P. cactorum. P. cryptogea and P. cactorum were confirmed by application of molecular methods. Nutritional requirements of P. cactorum and P. citrophthora were studied. Most Nitrogen sources stimulated the mycelial growth of P. cactorum to a higher extend, and reduced the colony size ofP. citrophthora. Different Carbon sources were utilized well by P. cactorum, and only saccharose and maltose had a stimulating effect on the mycelial growth of P. citrophthora. MgSO4.7H20 was the preferred sulfur source for both fungi, as L-cysteine and L-methionine only for P. cactorum. Phytophthora infection leads to physiological changes in the.host plant tissues. The tendency traced out is： about disorders in the amino acid metabolism, increase in the total sugars and slight reduction of the cellulose content. The total nitrogen, phosphorus and potassium content are reduced and the calcium and magnesium are increased. Photosynthesis of inoculated plants was suppressed and transpiration was increased.