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.

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。

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.

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。

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...

铜、砷及其复合污染对黄豆(Glycine max)影响的初步研究

通过水培实验研究了Cu、As单一及复合污染对黄豆种子萌发、幼苗生长及部分酶活性的影响 .结果表明 ,Cu、As污染明显抑制黄豆种子萌发、幼苗生长 ,对黄豆种子萌发时的呼吸强度、蛋白酶活性有显著的抑制作用 ,且随着Cu、As浓度的增加 ,抑制作用增强 ,呈负相关 ;而POD活性则随污染物浓度的增加而增加 ,呈正相关 .Cu、As污染共同存在时 ,随二者投加比例不同出现不同程度的拮抗效应 。

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.

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.

利用中国秋大豆(Glycine max(L.) Merr)筛选SSR核心位点的研究

选择中国秋大豆为试验材料 ,对基因组DNA进行SSR标记筛选和鉴定。经过 2 0 0个位点在琼脂糖胶上初筛和 96个位点在变性聚丙烯酰胺胶上复鉴 ,选出 6 0个位点 ,这些位点具有以下特点 :(1)分布在大豆 2 0个整合遗传连锁群 ,相邻位点间平均遗传距离在 5 0cM左右。除连锁群C2 、O上分别有 5个位点 ,G、K、M上分别有 2个位点外 ,其余 15个连锁群均分布有 3个位点 ;(2 )与 96个位点在 80份秋大豆种质检测到种质间遗传关系达到极显著相关 (r =0 .910 ) ;(3)在 80份秋大豆初选核心种质中表现出较高多态性 ,平均每个位点等位变异数为 9.3,多态性信息含量 (PIC)值为 0 .773;(4)在检测的秋大豆绝大多数种质基因组中 ,均为单一拷贝的位点 ,具有较高特异性 ;(5 )在相同的PCR扩增条件下 ,同一位点不同等位变异间易于识别且扩增强度较为一致。这套SSR核心位点的确定为中国大豆核心种质的构建奠定了基础。

不同茬口条件下的作物根渗出物对大豆胞囊线虫(Heterodera glycines)卵孵化影响

取北方5种作物轮作茬口和休闲土壤种植火豆、玉米、小麦、亚麻和甜菜,出苗后22 d制备作物的根渗出物,观察其对大豆胞囊线虫卯孵化的影响。试验表明,麦豆麦迎茬的甜菜根渗出物在后期大豆胞囊线虫(SCN)的卵孵化率最高,豆麦米轮作茬的大豆根渗出物9 d后SCN的卵孵化率明显高于其它作物,米豆米茬口的玉米根渗出物SCN的卵孵化率较高,豆麦豆迎茬甜菜根渗出物SCN的卵孵化率明显高于所有供试作物,12年大豆连作茬甜菜从一开始卵孵化率就高于对照和其它4种作物,休闲区的5种作物根渗出物也有对SCN卵孵化的刺激作用。

灰布支黑豆对大豆孢囊线虫(Heterodera glycines)14号小种的抗性遗传

大豆孢囊线虫（Ｈｅｔｅｒｏｄｅｒａ ｇｌｙｃｉｎｅｓ）是危害大豆生产的世界性病害。山西省兴县“灰布支黑豆”是对目前我国鉴定的所有流行小种表现出免疫或高抗的重要抗源。利用目前国际通用的一套鉴别寄主和小种划分标准，通过人工接种的方法，确定了１４号是北京马连洼中国农业科学院植物保护研究所实验站土壤中大豆孢囊线虫群体的主导小种。用敏感的栽培品种“冀豆７号”作母本，与灰布支黑豆杂交，采用人工接种的方法，对后代群体进行大豆孢囊线虫１４号小种的抗性鉴定。Ｆ１的２个单株都表现出抗性。随机取２个单株的Ｆ２代群体，分别测定每个群体的１１６和７８个单株。每个群体都表现出４３抗：２１感的分离比例，支持兴县灰布支黑豆对大豆孢囊线虫１４号小种的抗性是由３对基因控制、一对隐性基因对两对显性基因的上位和两对显性基因互补作用的遗传假设。随机取Ｆ３代的３０个株系，每个株系随机测定１０～１５个单株。１９抗：３８分离：７感的株系间分离比确认上述的遗传假设是正确的。

栽培大豆(Glycine max)与野生大豆(G.soja)解剖学的比较研究——Ⅰ.雄配子体的发育

本文对栽培大豆(Glycine max)和野生大豆(G.soja)雄配子体的发育和同步性及精子发生进行了比较研究。大豆和野生大豆花粉母细胞的减数分裂是属于同时型的,小孢子的发育过程基本相同。花粉粒的发育在同一花药中基本上是同步的,在同花中九个连体花药的花粉粒的发育也基本上是同步的,而单个花药中花药粉粒的发育要稍落后于九个连体花药中的花粉粒。大豆的两个精子是在花粉管和花粉粒中形成的,即二、三细胞型;野生大豆的两个精子是在花粉管中形成的即二细胞型。本文讨论了这两种植物精子发生途径的类型及分类学上的意义。

中国栽培大豆(Glycine max (L.) Merr.)微核心种质的群体结构与遗传多样性

【目的】评价中国栽培大豆微核心种质的群体结构和遗传多样性水平,为拓宽大豆遗传基础、发掘优异基因、改良大豆品种提供理论依据。【方法】利用大豆20个连锁群上的100个SSR位点,对来自全国28个省补充完善的248份栽培大豆微核心种质进行SSR遗传多样性及群体结构分析;采用PowerMarker Version 3.25软件统计等位变异数、平均等位变异数、多态性信息量(PIC值)及亚群特有等位变异数等参数;基于遗传距离建立了栽培大豆微核心种质的无根Neighbor-Joining树;用Structure2.2软件对微核心种质的群体结构进行评价。【结果】100个SSR位点在248份材料中共检测出等位变异1460个,每个位点变异范围为2—33个,平均为14.6个,每个位点PIC值变异范围为0.158—0.932,平均为0.743。基于模型的群体结构分析显示,依据LnP(D)无法判断最佳K值(群组数),但通过计算系数ΔK发现,K=3为微核心种质的最佳群体结构。结合种质的生态类型及品种类型分析发现,地理来源相同的种质具有聚在一起的倾向,但来源相同的种质也有分在不同组的情况。不同生态类型及品种类型间均存在较多的互补等位变异和特有等位变异。【结论】中国栽培大豆微核心种质具有丰富的遗传多样性,可以用来拓宽大豆品种遗传基础;不同生态类型及品种类型间存在较多的互补及特有等位变异,是种质创新及品种改良的物质基础;栽培大豆微核心种质存在明显的群体结构,为微核心种质在育种中的直接或间接利用提供了理论依据。

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.

高盐碱胁迫下野生大豆(Glycine soja)体内离子积累的差异

在总含量3%高盐碱原土盆栽条件下,对搜集于津唐盐碱地895份野生大豆材料进行全生育期耐盐碱性鉴定。通过测定203株凋亡植株及收获植株茎、叶中Na+、Cl-、K+、Ca2+、Mg2+的含量,分析高盐碱胁迫下野生大豆植株离子含量的分布及野生大豆植株凋亡和成熟时体内离子积累程度,并探讨耐盐碱型野生大豆的耐性机制。结果表明,野生大豆植株在营养生长期间,盐碱胁迫致死植株茎、叶无机离子含量在不同存活时间组间并未达显著水平,Na+和Cl-积累达到一定含量即出现凋亡现象,致死植株茎中Na+和Cl-离子范围分别为3.239%～4.682%和4.639%～6.328%,叶中分别为1.754%～2.349%和4.126%～5.073%;能够存活到成熟的耐盐碱型野生大豆植株茎叶Na+和Cl-含量存在低、中、高3种类型。高耐型野生大豆茎、叶平均Na+和Cl-含量显著低于低耐盐型,且茎中K+和叶中Ca2+和Mg2+含量较高。在高耐型野生大豆植株茎叶中也存在Na+和Cl-离子含量高水平和低水平两种类型,推测野生大豆可能存在两种耐盐碱机制,其一为高耐受性,其二为低吸收性。

野生大豆(Glycine. soja)抗锈鉴定

大豆锈病近年来已发展成为影响大豆生产的世界性病害。筛选抗锈资源是开展抗病育种的基础,从野生大豆中发掘抗锈资源有利于拓宽抗锈育种的遗传基础。采用离体叶片定量接种方法对513份野生大豆(Glycine so-ja)进行抗锈鉴定,得到抗病资源1份,其余均为感病资源。

国家基因库野生大豆(Glycine soja)资源最近十年考察与研究

总结了国家种质库最近10年野生大豆搜集进展和研究概况。我国野生大豆资源在1979-1982年间搜集并保存在国家基因库的为5939份;1996-2000年搜集了600份;2001-2010最近10年补充搜集了全国17个省(市、区)、318个县(市、旗)、930个乡镇(农场)。其中新搜集县市178个,共收集典型野生大豆资源1979份,野生大豆收集样品新增资源33.3%。

渤海湾津唐沿海野生大豆(Glycine soja)种群高盐碱胁迫反应

以渤海湾津唐沿海地带895份野生大豆种群在高盐碱(3%)环境下的全生育期鉴定试验,分析不同种群(株系)的生长发育动态以及对盐碱土壤生态适应性的分化。结果表明,出苗阶段存在3种耐盐碱能力类型:耐高盐碱胁迫发芽出苗;耐低盐碱胁迫发芽出苗;盐胁迫不能发芽出苗。在高盐胁迫下营养生长早期阶段种群死亡率最高,中期以后相对降低,成熟期死亡率较低。种群植株形态建成和生殖生长受到高盐胁迫的强烈抑制,表现早期严重而后期较轻。植株和产量性状的受害程度由大到小依次为:单株的地上干物质重、单株产量、种子数、百粒重和收获指数。株系间变异系数从大到小依次为:单株产量、种子数、荚数、收获指数、地上干物质重、百粒重。研究揭示该地域种群内对盐碱土壤有很高的适应性分化,存在高耐盐碱生态型。

大豆遗传图谱的构建和抗胞囊线虫(Heterodera glycines Ichinohe)的QTL分析

大豆胞囊线虫1号和4号生理小种是黄淮地区的优势小种，ZDD2315是我国特优抗源。本文旨在定位ZDD2315对1号和4号生理小种抗性的QTL。试验以Essex为母本，ZDD2315为父本和轮回亲本，创建了一个包含114个单株的Bc。群体。采用250个SSR标记和1个形态标记通过MAPMAKER3．0构建了包含25个连锁群的遗传图谱，覆盖大豆基因组2963．5cM，平均每个连锁群上10．0个标记，标记平均间距11．8cM。采用Win QTL Cartographer Version 2．5复合区间作图法（CIM）检测到3个抗1号小种的QTL；其中rhgR1-1和rhgR1—2位于G连锁群的Sat_210～Sat_168和Sat_168～Sat_141区间，贡献率分别为22．4％和21.8％；rhgR1-3位于D2连锁群的Satt672～Satt413区间，贡献率6．2％；rhgR1-1和rhgR1—3分别与Sat_210和Satt672共分离。5个QTL与抗4号生理小种有关；其中rhgR4—1和rhgR4—-位于G连锁群的Satt275～Sat_210和Sat_168～Sat_141区间，贡献率分别为22．8％和28．9％；rhgR4—3和rhgR4—4位于H连锁群Satt442～Sat401和Sat_334～Satt181区间，贡献率分别为12．0％和10．5％；rhgR4—5位于L连锁群Satt652～Sat_301区间，贡献率5．9％；吨职4—2和rhgR4—5分别与Sat_168和Satt652共分离。不同遗传体系控制ZDD2315对1号和4号小种的抗性。抗1号和4号生理小种的主要QTL位于G连锁群的相近区段，且具有较大贡献率，通过标记辅助选择有可能育成兼抗两小种的品种。