Dietary glycine supplementation enhances syntheses of creatine and glutathione by tissues of hybrid striped bass(Morone saxatilis ♀ × Morone chrysops ♂) fed soybean meal-based diets

Background We recently reported that supplementing glycine to soybean meal-based diets is necessary for the optimum growth of 5-to 40-g(Phase-I)and 110-to 240-g(Phase-II)hybrid striped bass(HSB),as well as their intestinal health.Although glycine serves as an essential substrate for syntheses of creatine and glutathione(GSH)in mammals(e.g.,pigs),little is known about these metabolic pathways or their nutritional regulation in fish.This study tested the hypothesis that glycine supplementation enhances the activities of creatine-and GSH-forming enzymes as well as creatine and GSH availabilities in tissues of hybrid striped bass(HSB;Morone saxatilis♀×Morone chrysops♂).Methods Phase-I and Phase-II HSB were fed a soybean meal-based diet supplemented with 0%,1%,or 2%glycine for 8 weeks.At the end of the 56-d feeding,tissues(liver,intestine,skeletal muscle,kidneys,and pancreas)were collected for biochemical analyses.Results In contrast to terrestrial mammals and birds,creatine synthesis occurred primarily in skeletal muscle from all HSB.The liver was most active in GSH synthesis among the HSB tissues studied.In Phase-I HSB,supplementation with 1%or 2%glycine increased(P<0.05)concentrations of intramuscular creatine(15%–19%)and hepatic GSH(8%–11%),while reducing(P<0.05)hepatic GSH sulfide(GSSG)/GSH ratios by 14%–15%,compared with the 0-glycine group;there were no differences(P>0.05)in these variables between the 1%and 2%glycine groups.In Phase-II HSB,supplementation with 1%and 2%glycine increased(P<0.05)concentrations of creatine and GSH in the muscle(15%–27%)and liver(11%–20%)in a dose-dependent manner,with reduced ratios of hepatic GSSG/GSH in the 1%or 2%glycine group.In all HSB,supplementation with 1%and 2%glycine dose-dependently increased(P<0.05)activities of intramuscular arginine:glycine amidinotransferase(22%–41%)and hepaticγ-glutamylcysteine synthetase(17%–37%),with elevated activities of intramuscular guanidinoacetate methyltransferase and hepatic GSH synthetase and GSH reductase in the 1%or 2%glycine group.Glycine supplementation also increased(P<0.05)concentrations of creatine and activities of its synthetic enzymes in tail kidneys and pancreas,and concentrations of GSH and activities of its synthetic enzymes in the proximal intestine.Conclusions Skeletal muscle and liver are the major organs for creatine and GSH syntheses in HSB,respectively.Dietary glycine intake regulates creatine and GSH syntheses by both Phase-I and Phase-II HSB in a tissue-specific manner.Based on the metabolic data,glycine is a conditionally essential amino acid for the growing fish.

Hypomethylation of glycine dehydrogenase promoter in peripheral blood mononuclear cells is a new diagnostic marker of hepatitis B virus-associated hepatocellular carcinoma

Background: Glycine dehydrogenase(GLDC) plays an important role in the initiation and proliferation of several human cancers. In this study, we aimed to detect the methylation status of GLDC promoter and its diagnostic value for hepatitis B virus-associated hepatocellular carcinoma(HBV-HCC). Methods: We enrolled 197 patients, 111 with HBV-HCC, 51 with chronic hepatitis B(CHB), and 35 healthy controls(HCs). The methylation status of GLDC promoter in peripheral mononuclear cells(PBMCs) was identified by methylation specific polymerase chain reaction(MSP). The mRNA expression was examined using real-time quantitative polymerase chain reaction(q PCR). Results: The methylation frequency of the GLDC promoter was significantly lower in HBV-HCC patients(27.0%) compared to that in CHB patients(68.6%) and HCs(74.3%)( P < 0.001). The methylated group had lower alanine aminotransferase level( P = 0.035) and lower rates of tumor node metastasis(TNM) Ⅲ/Ⅳ( P = 0.043) and T3/T4( P = 0.026). TNM stage was identified to be an independent factor for GLDC promoter methylation. GLDC mRNA levels in CHB patients and HCs were significantly lower than those in HBV-HCC patients( P = 0.022 and P < 0.001, respectively). GLDC mRNA levels were significantly higher in HBV-HCC patients with unmethylated GLDC promoters than those with methylated GLDC promoters( P = 0.003). The diagnostic accuracy of alpha-fetoprotein(AFP) combined with GLDC promoter methylation for HBV-HCC was improved compared with that of AFP alone(AUC: 0.782 vs. 0.630, P < 0.001). In addition, GLDC promoter methylation was an independent predictor for overall survival of HBV-HCC patients( P = 0.038). Conclusions: The methylation frequency of GLDC promoter was lower in PBMCs from HBV-HCC patients than that from patients with CHB and HCs. The combination of AFP and GLDC promoter hypomethylation significantly improved the diagnostic accuracy of HBV-HCC.

A novel procedure for identifying a hybrid QTL-allele system for hybrid-vigor improvement, with a case study in soybean(Glycine max)yield

“Breeding by design” for pure lines may be achieved by construction of an additive QTL-allele matrix in a germplasm panel or breeding population, but this option is not available for hybrids, where both additive and dominance QTL-allele matrices must be constructed. In this study, a hybrid-QTL identification approach, designated PLSRGA, using partial least squares regression(PLSR) for model fitting integrated with a genetic algorithm(GA) for variable selection based on a multi-locus, multi-allele model is described for additive and dominance QTL-allele detection in a diallel hybrid population(DHP). The PLSRGA was shown by simulation experiments to be superior to single-marker analysis and was then used for QTL-allele identification in a soybean DPH yield experiment with eight parents. Twenty-eight main-effect QTL with 138 alleles and nine QTL × environment QTL with 46 alleles were identified, with respective contributions of 61.8% and 23.5% of phenotypic variation. Main-effect additive and dominance QTL-allele matrices were established as a compact form of the DHP genetic structure. The mechanism of heterosis superior-to-parents(or superior-to-parents heterosis, SPH) was explored and might be explained by a complementary locus-set composed of OD+(showing positive over-dominance, most often), PD+(showing positive partial-to-complete dominance, less often) and HA+(showing positive homozygous additivity, occasionally) loci, depending on the parental materials. Any locus-type, whether OD+, PD + and HA+, could be the best genotype of a locus. All hybrids showed various numbers of better or best genotypes at many but not necessarily all loci, indicating further SPH improvement. Based on the additive/dominance QTL-allele matrices, the best hybrid genotype was predicted, and a hybrid improvement approach is suggested. PLSRGA is powerful for hybrid QTL-allele detection and cross-SPH improvement.

Performance in Survival,Development and Reproduction of Aphis glycines Matsumura Virginoparae at High Temperatures

The soybean aphid,Aphis glycines Matsumura,is an important pest of soybean,which is native to Asia.In this study,A.glycines fed on soybean(AgFS)and A.glycines fed on wild soybean(AgFW)were reared at 25℃,27℃,29℃,31℃,33℃and 35℃,respectively,and some of the life parameters were determined.At temperature ranging from 25℃to 31℃,nymphs of AgFS and AgFW all developed into adults successfully.Only a few nymphs of AgFS and AgFW developed into adults at 33℃and no nymphs could develop into adults at 35℃.Lifespan,fecundity and body size of AgFS and AgFW adults all decreased gradually with temperatures increasing from 25℃to 33℃.At 25℃,the intrinsic rate of increase of AgFS was as big as that at 27℃,which was smaller than that at 29℃,but was bigger than that at 31℃.Intrinsic rate of increase of AgFW decreased gradually with temperatures increasing from 25℃to 31℃.Nymph stage duration of AgFW was longer than or as long as that of AgFS;adult lifespan of AgFW was shorter than or as long as that of AgFS.Adult fecundity,intrinsic rate of increase and adult body size of AgFW were all smaller than or as big as those of AgFS.It showed that AgFS and AgFW both survived and developed well at temperature ranging from 25℃to 31℃,and AgFW was more adaptive to low temperatures.These results were important to study the adaptability of A.glycines to high temperatures and for predicting its dynamics in the temperature keeping rising region.

大豆对胞囊线虫(Heterodera glycines Ichinohe)1号和4号生理小种抗性的遗传分析

大豆胞囊线虫(Heterodera glycinesIchinohe)是我国大豆的全国性主要病害之一。1号和4号生理小种是黄淮地区的优势小种。以Essex×ZDD2315、Peking×ZDD2315、PI88788×ZDD2226、Peking×ZDD2226的P1、P2、F1、BC1F2为材料,用主基因+多基因混合遗传模型分析大豆对胞囊线虫1号和4号生理小种抗性的遗传机制。结果表明,ZDD2315、ZDD2226对1号生理小种的抗性受主效基因控制,未发现多基因效应,且与Peking存在相同的抗病基因;抗性遗传表现组合特异性,Essex×ZDD2315组合为3对加性主基因遗传模型,主基因遗传率72.02%,PI88788×ZDD2226组合为2对显性上位主基因遗传模型,主基因遗传率62.33%。对4号生理小种的抗性为主基因+多基因混合遗传模型,Essex×ZDD2315、Peking×ZDD2315、PI88788×ZDD2226等3个组合为3对主基因+多基因遗传模型,主基因遗传率分别为67.76%7、2.46%和53.25%,多基因遗传率分别为24.48%、21.31%和35.77%;Peking×ZDD2226表现为2对主基因遗传模型,主基因遗传率45.40%。抗性基因表现为隐性,育种上可以在早代选择。培育多抗品种应以抗4号生理小种为主要目标进行基因聚合。

干旱和复水对大豆(Glycine max)叶片光合及叶绿素荧光的影响

选用河南省大面积种植的大豆品种豫豆29作为实验材料,通过研究逐步干旱和旱后复水条件下大豆叶片光合、叶绿素荧光等指标随土壤水分的动态变化规律,以期为大豆的水分高效利用提供理论依据。研究发现,在土壤相对含水量高于46.5%时,虽然随着土壤相对含水量的下降,豫豆29仍可以保持它的叶片水分状态;豫豆29的叶片净光合速率在土壤水分中等条件下最大,在土壤相对含水量为64.3%时,它比对照组高出11.2%(P<0.01);在实验的第3d,处理组的土壤相对含水量降为46.5%,叶片水势与对照组相比降低了7.2%(P>0.05),净光合速率为对照组的89.6%(P<0.05),但气孔导度却迅速下降为对照组的44.7%(P<0.01),这说明与叶片的光合和水分状况相比,豫豆29的气孔对土壤水分的匮缺更加敏感。复水后,豫豆29叶片的水势、净光合速率、气孔导度和叶绿素荧光等值都可以得到迅速的恢复,并在实验的最后接近对照组的水平,这表明豫豆29的叶片光合在水分胁迫解除后有迅速恢复的能力。

大豆属Glycine亚属植物花粉形态研究

对大豆属Glycine亚属中10个种的花粉进行了光学显微镜和扫描电镜的观察.发现多倍体材料的花粉大于二倍体的.并发现Glycine亚属诸种花粉的形状、萌发孔、外壁纹饰均存明显差异.

Studies of Glycine Betaine on Physiology of Two Varieties of Pumpkin Seedlings under NaCl Stress

[Objective] The research aimed to discuss the relationship between Glycine betaine and the salt-tolerance mechanism of different pumpkins and confirm the mechanism of betaine in salt-tolerance physiology.[Method] Taking the seedlings of C.ficifolia and Qingli pumpkin as test materials,the effects of glycine betaine on the cell membrane permeability,MDA and root activity of two varieties of pumpkin seedlings under 300 mmol/L NaCl stress were studied.[Result] At suitable concentrations,glycine betaine could d...

Cd Distribution and Subcellular Localization in Leaf and Its Effects on Growth of Soybean(Glycine max) Seedlings

In order to investigate Cd accumulation, subcellular distribution, and local-ization in soybean seedlings leaves, soybean seedlings were cultivated in solution containing different concentrations of Cd. The results showed that most Cd associ-ated with the cellwal s and soluble fractions, and a minor part of Cd presented in mitochondria fractions, nuclear and chloroplast fractions, especial y exposure to high Cd concentrations. Under 20.00 mg/L Cd stress, Cd subcellular distribution fol owed a sequence as： soluble fractions （55.00%）＆gt;cellwal s （30.0%）＆gt;mitochondria fractions （8.21%）＆gt;nuclear and chloroplast fractions （6.79%）. Deposited Cd black particles were observed in cellwal s, chloroplasts, nuclei, and vacuoles through electrical microscope slice. This fact indicated that the cellwal s of soybean leaves were the first protecting organel es from Cd toxicity, and the cellwal s and soluble fractions were the main place for Cd storage. Due to Cd accumulated in the organel es, the intercellular space was enlarged and the subcellular structure was damaged, especial y for the chloroplasts.

Discussion on the Problem of Salt Gland of Glycine soja

Glycine soja Sieb. et Zucc. plants living in saline soil in three provinces of China were treated with different salinity concentrations under different laboratory culture conditions (including solution, sand and field cultivation). The attachment shape and distribution on the surface of stalk and leaf of G. soja plants were observed with scanning electron microscopy (SEM), and the ultrastructure of glandular hair with transmission electron microscopy (TEM). Na+ and Cl- contents in the secretion of the leaf surface and inside the leaf of G. soja subjected to different treatments were measured. The Na+ relative contents in glandular cells, epidermal cells and mesophyllous cells of leaves under different salinities were determined by X-ray microanalysis. Results show that only glandular and epidermal hair exist on the surface attachments of leaves and stalks of G. soja plants. These glandular hair were similar in shape to some salt glands of Gramineae halophytes, and they attached to the vein on the leaf surface. The cell structure of the glandular hair showed the characteristics of common salt glands, such as big vacuoles, dense cytoplasm, a great deal of mitochondria, chloroplast, plasmodesmata and thicker cell walls, etc. The results of Na+ and Cl- contents in the leaf secretion and inside the leaf showed that the glandular hair executed the function of salt-secretion, and when treated with the salt gland inhibitor the salt-secretion process was inhibited. As a result, Na+ and Cl- were mainly accumulated inside G. soja leaves. The results of Na+ X-ray microanalysis under different salinities proved that the three cells of the glandular hair, especially the top cell, possessed strong competence for Na+ accumulation. Above all, the glandular hair were the salt gland, and no other kind of salt glands were found on G. soja plants. The secreting mechanism of the salt gland was also discussed.

Isolation and Characterization of a Mitochondrial atp6 Gene from Soybean (Glycine max)

Mitochondrial ATPase (mtATPase) complex plays vital roles in higher plants. It consists of a few subunits. In the present study, a new copy of the mtATPase subunit 6 (EC 3.6.1.34) gene (atp6) was cloned and characterized from Glycine max (L.) Merr., which has the shortest opening reading frame of 223 amino acids in all organisms examined and designated as the atp6 copy3 (atp6_3). PCR amplifications of the atp6_3 from 9 soybean cultivars combined with sequencing analysis suggested its wide occurrence in G. max . RFLP analysis of a RILs population implied that paternal inheritance of the atp6_3 might occur in G. max at undetermined frequency. Under salicylic acid (SA)_treated condition, the expression of the atp6 gene was significantly inhibited. The possible role of this inhibition was discussed.

野生大豆(GlycinE Soja)抗蚜鉴定技术方法研究初报

目测分级鉴定野生大豆对大豆蚜的抗性是一种简便、有效的方法。被鉴定材料扣网接蚜可以大大提高蚜虫的发生量及鉴定结果的准确性。接虫虫源可以在栽培大豆上预先繁殖。野生大豆苗期及开花期对蚜虫危害的反应基本一致。

Glycine attenuates myocardial ischemia-reperfusion injury by inhibiting myocardial apoptosis in rats

Glycine is a well-documented cytoprotective agent.However,whether it has a protective effect against myocar-dial ischemia-reperfusion injury in vivo is still unknown.By using an open-chest anesthetized rat model,we found that glycine reduced the infarct size by 21% in ischemia-reperfusion injury rats compared with that in the vehicle-treated MI/R rats.The left ventricular ejection fraction and fractional shortening were increased by 19.11% and 30.98%,respectively,in glycine-treated rats.The plasma creatine kinase levels in ischemia-reperfusion injury rats decreased following glycine treatment.Importantly,administration of glycine significantly inhibited apoptosis in post-ischemia-reperfusion myocardium,which was accompanied by suppression of phosphorylated p38 mitogen-activated protein kinase and c-Jun NH2-terminal kinase,as well as the Fas ligand.These results suggest that gly-cine attenuates myocardial ischemia-reperfusion injury in vivo by inhibiting cardiomyocytes apoptosis.

Sampling Survey and Identification of Races of Soybean Cyst Nematode (Heterodera glycines Ichinohe) in Huang-Huai Valleys

Soybean cyst nematode （SCN Heterodera glycines Ichinohe） is one of the most important nationwide soybean diseases in China. A total of 38 soil specimens or locations in the area was sampled and tested for SCN races during 2001-2003 for the inspection of race distribution in Huang-Huai Valleys. A map of race distribution was constructed according to the data from both the present study and the published reports cited. Three areas, namely, the area of southeast to Jinan in Shangdong Province; the area of northern Henan Province and its border region to south of Hebei Province; and the area of Luohe, Zhoukou of Henan Province and Fuyang of Anhui Province mainly infested with Race 1 were identified. Race 4 was predominant in Shanxi Province, Beijing and the adjacent area of Henan, Shandong, and Anhui provinces, and the delta of Huanghe River in Shandong Province. Race 2 was mainly found in Liaocheng, Dezhou of Shangdong Province and Shijiazhuang of Hebei Province, and Jiaozuo and Huojia of Henan Province. Race 7 was distributed in the west part of Jiaodong Peninsula of Shandong Province and Kaifeng, Huaxian, Wenxian of Henan Province. Race 5 was found and scattered in Hebei and Henan Province. Race 9 was found in Shangqiu of Henan Province, which was reported for the first time in China. It can be seen that Race 1 and Race 4 were the two predominant races in Huang-Huai Valleys, and that research should focus on developing resistant cultivars of these races. There might exist other races in an area with some predominant races. The race substitution in the past decade was not obviously found, therefore, the results should be meaningful to future breeding for resistance to SCN in Huang-Huai Valleys.

Effects of Exogenous Glycine Betaine on Oxidation Metabolism in Cucumbers during Low-temperature Storage

[Objective] This study aimed to analyze the effects of different concentrations of glycine betaine（GB） on oxidation metabolism in cucumbers under low-temperature stress and to investigate the possible mechanism of low-temperature resistance in cucumber during low-temperature storage. [Method] Cucumber cultivar Zhongnong No.8 was treated with 0, 5, 10 and 15 mmol/L GB solutions for 15 min and stored at 4 ℃. Changes in oxidative metabolism-related parameters were observed. [Result] Increasing exogenous GB concentration could enhance GB content in cucumbers, decline lipoxygenase（LOX） activity, improve peroxidase（POD） and catalase（CAT） activities, remove effectively hydrogen peroxide（H2O2） and reduce the accumulation of malondialdehyde（MDA）. [Conclusion] Treating cucumbers with10 mmol/L GB exhibited the most remarkable effect.

Inheritance of the Anatomy--Morphological Structure of the Stalk by Interspecific Hybrids of the Glycine L.

The research focuses on the study of anatomical and morphological stalk structure ofsoya interspecific hybrids of the third generation （F3） between （Glycine max （L.） Merr.） and G. soja Sieb. et Zucc. in comparison with parent plant species. The parent plant species and interspecific hybrids were sowed and grew under similar conditions. The similarity of the anatomic structure of stalks of cultivated plants and Glycine soja （wild soya） proves the hypothesis the studied species have the same origin. However, the obtained results show the considerable degree of phylogenetic dissociation between the studied soya species. Interspecific hybrids inherit from G. soja the ability to high intensive growth. The G. soja use in practical selective breeding is of great interest.

Effects of salinity on activities of H^+-ATPase, H^+-PPase and membrane lipid composition in plasma membrane and tonoplast vesicles isolated from soybean(Glycine max L.) seedlings

The effects of NaCl stress on the H +-ATPase, H +-PPase activity and lipid composition of plasma membrane(PM) and tonoplast(TP) vesicles isolated from roots and leaves of two soybean cultivars(Glycine max L.) differing in salt tolerance(Wenfeng7, salt-tolerant; Union, salt-sensitive) were investigated. When Wenfeng7 was treated with 0.3%(W/V) NaCl for 3 d, the H +-ATPase activities in PM and TP from roots and leaves exhibited a reduction and an enhancement, respectively. The H +-PPase activity in TP from roots also increased. Similar effects were not observed in roots of Union. In addition, the increases of phospholipid content and ratios of phospholipid to galactolipid in PM and TP from roots and leaves of Wenfeng7 may also change membrane permeability and hence affect salt tolerance.

Mitigative Effect of La on Glycine max seedlings under Pb-Cd Compound Pollution

Mitigative effect of La on Glycine max seedlings under combined Pb and Cd pollution was studied through pot culture experiment. The results show that the growth and metabolism of Glycine max seedlings are inhibited by the solution with 500 mg.L-1 Pb + 100 mg.L-1 Cd. When 30 mg. L-1 LaCl3 is used to spray Glycine max seedlings once, the injury effect of combined Pb and Cd pollution is reduced. The experiment proves that the effect is related to La which can raise photosynthetic rate, chlorophyll content and activity of nitrate reductase, and reduce cell membrane permeability, content of Pb and Cd, and keep TTC reduction ability of Glycine max seedling.

Mapping and validation of a dominant salt tolerance gene in the cultivated soybean(Glycine max) variety Tiefeng 8

Salt is an abiotic stress factor that strongly affects soybean growth and production. A single dominant gene has been shown to confer salt tolerance in the soybean cultivar Tiefeng 8.The objective of the present study was to genetically map the salt-tolerance gene in an F2:3population and a recombinant inbred line(RIL) population derived from a cross between two cultivated soybeans, Tiefeng 8(tolerant) and 85-140(sensitive). The F2:3families and RILs were treated with 200 mmol L-1Na Cl to evaluate salt tolerance. The F2:3population showed 1(42 tolerant): 2(132 segregating): 1(65 sensitive) segregation, indicating a single dominant gene for salt tolerance in Tiefeng 8. A sequence-characterized amplified region(SCAR) marker from a previously identified random amplified polymorphic DNA(RAPD)marker and four insertion/deletion polymorphism(In Del) markers were developed within the mapping region. Using these markers along with SSR markers, the salt-tolerance gene was mapped within 209 kb flanked by SCAR marker QS08064 and SSR marker Barcsoyssr_3_1301 on chromosome 3. Three markers that cosegregated with the salt tolerance gene and SCAR marker QS08064 were used to genotype 35 tolerant and 23 sensitive soybean accessions. These markers showed selection efficiencies of 76.2% to94.2%. The results indicate that these markers will be useful for marker-assisted breeding and facilitating map-based cloning of the salt tolerance gene in soybean.

Overaccumulation of glycine betaine makes the function of the thylakoid membrane better in wheat under salt stress

Wheat(Triticum aestivum L.) lines T1, T4, and T6 were genetically modified to increase glycine betaine(GB) synthesis by introduction of the BADH(betaine aldehyde dehydrogenase, BADH)gene from mountain spinach(Atriplex hortensis L.). These transgenic lines and WT of wheat(T. aestivum L.) were used to study the effect of increased GB synthesis on wheat tolerance to salt stress. Salt stress due to 200 mmol L-1Na Cl impaired the photosynthesis of the four wheat lines, as indicated by declines in net photosynthetic rate(Pn), stomatal conductance(Gs),maximum photochemical efficiency of PSII(Fv/Fm), and actual photochemical efficiency of PSII(ФPSII) and an increase in intercellular CO2concentration(Ci). In comparison with WT, the effect of salinity on the three transgenic lines was mild. Salt stress caused disadvantageous changes in lipids and their fatty acid compositions in the thylakoid membrane of the transgenic lines and WT. Under salt stress, the three transgenic lines showed slightly higher chlorophyll and carotenoid contents and higher Hill reaction activities and Ca2+-ATPase activity than WT. All the results suggest that overaccumulation of GB resulting from introduction of the BADH gene can enhance the salt tolerance of transgenic plants, especially in the protection of the components and function of thylakoid membranes, thereby making photosynthesis better. Changes in lipids and fatty acid compositions in the thylakoid membrane may be involved in the increased salt stress tolerance of the transgenic lines.