高温胁迫下紫花苜蓿多胺含量与耐热性的关系

我国缺乏适宜在南方高温地区生长的优质紫花苜蓿(Medicago sativa)品种,研究紫花苜蓿耐热性具有重要意义。多胺与植物逆境胁迫响应密切相关。本试验以高秋眠级紫花苜蓿品种‘赛迪10’、低秋眠级紫花苜蓿品种‘阿尔冈金’和适应云南干热河谷的‘德钦’紫花苜蓿为研究材料,研究腐胺、亚精胺、精胺含量在胁迫温度35℃/30℃(白天/夜晚)条件下105天内的变化,分析多胺与紫花苜蓿耐热性之间的关系。结果表明,紫花苜蓿的腐胺和精胺含量在高温胁迫下先升后降,期间低秋眠级品种的腐胺和精胺含量较高,高秋眠级品种的精胺含量开始下降的时间比低秋眠级品种早大约21天;紫花苜蓿的亚精胺含量在高温胁迫下变化有所不同,高秋眠级品种的亚精胺含量会有所增加,而低秋眠级品种则显著下降,累计增长率分别为16.81%和-26.21%。‘德钦’紫花苜蓿与‘阿尔冈金’同为低秋眠级紫花苜蓿品种,但在高温胁迫下其多胺含量的变化却与高秋眠级品种‘赛迪10’一致。高温胁迫下德钦紫花苜蓿的耐热性可能与较高的亚精胺含量和较低的腐胺和精胺含量有关,是其能在干热河谷生存的原因之一。     

高温胁迫下‘德钦’紫花苜蓿内源激素的变化

为探析‘德钦’紫花苜蓿(Medicago sativa L.‘Deqin’)特异耐热性的原因,本试验以高秋眠级耐热型‘赛迪10’和低秋眠级热敏型‘阿尔冈金’为对照,比较35℃/30℃(昼温/夜温)高温胁迫下‘德钦’叶片IAA,CTK,GA_3和ABA 4种内源激素的变化。结果表明,具有增强植物耐热性的ABA在‘赛迪10’中含量最高并且在高温胁迫初期急剧增加,耐热性与ABA作用相反的GA_3在‘赛迪10’中始终维持较低的水平。‘德钦’的ABA和GA_3变化趋势与‘赛迪10’相似,其ABA在高温胁迫第7d小幅增加(4.16%),也能维持相对较高的水平,但GA_3累计增加量小于‘阿尔冈金’。延缓叶片衰老的IAA和CTK在‘德钦’中的含量高,累计变化率分别为74.23%和21.6%,显著高于‘阿尔冈金’和‘赛迪10’。‘德钦’特异性耐热的原因可能与其响应高温胁迫相对较早和高水平ABA有关,且IAA和CTK含量较高,GA_3含量和累计增加量较低。     

紫花苜蓿水分利用效率对水分胁迫的响应及其机理

采用盆栽试验,测定了3种紫花苜蓿品种的光合(Pn)、蒸腾(Tr)、胞间CO₂浓度(Ci)、气孔密度和叶片碳同位素组成(δ¹³C)等,探讨不同紫花苜蓿品种在水分胁迫下的水分利用效率(WUE)的变化。结果显示,陇东苜蓿和新疆大叶苜蓿叶片WUE在50%田间持水量(FWC)水分胁迫下最高,在75%FWC下最低,而阿尔冈金苜蓿则相反;以日为尺度,各品种在25%FWC下WUE最高,在75%和50%FWC下WUE大小则因品种不同而表现不同;Pn/Ci随水分胁迫增加而下降,而气孔密度上升;δ¹³C值也随水分胁迫的增加而增加。上述结果表明,水分胁迫可提高紫花苜蓿水分利用效率,但品种间有差异;气孔因素和叶片羧化效率共同影响紫花苜蓿的水分利用。     

种子萌发期紫花苜蓿秋眠级与耐热性的关系

为了探明高温胁迫对不同秋眠级紫花苜蓿种子萌发的影响,在25和35℃条件下,对14个不同秋眠级紫花苜蓿品种种子进行萌发试验,以发芽率、发芽指数、活力指数、胚根鲜重和发芽势为指标,结合协方差分析、模糊隶属函数和聚类分析统计方法,对各紫花苜蓿品种萌发期耐热性进行分析和综合评价。结果表明：高温胁迫对不同秋眠级紫花苜蓿的种子萌发有影响,高秋眠级品种赛迪10的耐热最强,其发芽指数、活力指数、胚根鲜重、发芽势和发芽率在14个参试品种中最高,分别为67.78、66.70、0.98g、99.33%和92.89%;聚类分析的结果表明,14个品种萌发期耐热性可划分为3类,耐热性较强的品种为标秆、WL903、1075、三得利、赛迪10和赛阔,耐热性较弱的品种为Power、猎人河和阿尔冈金,其余品种耐热性一般;在种子萌发阶段,高秋眠级类型的紫花苜蓿品种表现较强的耐热性,低秋眠级类型的品种耐热性较弱。种子活力指数可作为评价紫花苜蓿萌发期耐热性强弱的有效指标。     

水分胁迫和复水对紫花苜蓿幼苗叶绿素荧光特性的影响

利用PEG-6000模拟水分胁迫(ψs=-0.2MPa,胁迫时间:48 h,复水48 h),研究了紫花苜蓿(Medicago sativa L)品种陇东苜蓿和阿尔冈金叶片叶绿素荧光特性和光合色素含量的变化,旨在探讨紫花苜蓿水分胁迫条件下的光合反应机制.结果表明:光化学淬灭系数(qP)和PSII线性电子传递的有效量子产额(YIELD)以及非光化学淬灭系数(qN)和表观光合电子传递效率(ETR)在胁迫前后随光合有效辐射(PAR)的变化规律分别可用公式:Y=aLn(X) b(Y:qP或YIELD,X:PAR,X≠0)和Y=aX2 bX c(Y:qN或ETR,X:PAR)来表示;在受到干旱胁迫后,PSII最大光能转化效率(Fv/Fm)、PSII潜在活性(Fv/Fo)和光合色素含量均明显低于对照,在任意光强下的qP、YIELD和ETR值也极显著低于对照的相应值,qN值则极显著高于对照的相应值;同时,和对照相比,qN和ETR光响应曲线顶点所对应的光强值在受到胁迫后亦明显下降,表明光抑制初始点的降低和最大光保护能力的减弱,甚至光抑制的提前到来.干旱胁迫使得PSⅡ反应中心结构和功能遭到破坏而部分关闭,光合电子传递受阻,光能利用与光化学转化与能力下降,吸收的光能更多的以热能形式耗散.复水后,虽然有光保护机制的存在,但除陇东苜蓿光合色素含量及阿尔冈金类胡萝卜素外,各参数均未能恢复到对照水平,因此旱后复水不能完全解除干旱对PSII反应中心所带来的伤害.紫花苜蓿幼苗对干旱胁迫较为敏感.供试品种相比,陇东苜蓿耐旱性优于阿尔冈金.     

水分胁迫对紫花苜蓿根系吸水与光合特性的影响

变水条件下(利用PEG-6000模拟水分胁迫48 h,ψs=0.2MPa,之后复水48 h),测定紫花苜蓿(Medicago sattva L,品种阿尔冈金和陇东苜蓿)幼苗根系水力学导度与光合参数变化规律,旨在研究变水条件下紫花苜蓿光合作用的响应机制、植株水分吸收能力的变化规律及地上与地下部可能的相关关系.结果表明:干旱胁迫使得紫花苜蓿根系水力学导度(Lpr)受到显著影响.随着水分胁迫时间的延长,根系吸水能力呈现快速--缓慢下降趋势,即根系水力学导度逐渐下降,气孔导度(Gs)、蒸腾速率(Tr)、净光合速率(Pn)和叶片水势亦随之而显著降低,胞间二氧化碳浓度(Ci)先随之降低但最终上升而累积.复水后,根系水导呈现缓慢--慢速的恢复趋势,Pn、Gs、Tr、叶片水势随着根系水导速率的增加而逐渐恢复;Ci则随复水时间的延长而逐渐下降.但各参数除Ci外,均没有恢复到胁迫前水平.紫花苜蓿根系水导与光合参数在复水后的恢复程度说明,紫花苜蓿对干旱逆境的抵御与适应能力相对较弱,但陇东苜蓿对水分胁迫的忍耐能力强于阿尔冈金.     

干旱和高温胁迫下紫花苜蓿PEPC酶活性变化及其基因序列分析

紫花苜蓿(Medicago sativa L.)是一种分布极其广泛的多年生优质豆科牧草，分析干热胁迫下其磷酸烯醇式丙酮酸羧化酶(Phosphoenolpyruvate carboxylase, PEPC)酶活性变化及PEPC蛋白的生物信息学。本研究以‘阿尔冈金’紫花苜蓿和云南德钦县野生紫花苜蓿为材料，测定其在干旱胁迫和高温胁迫下的PEPC酶活性的变化，同时扩增了PEPC基因，并对PEPC蛋白进行了生物信息学分析和系统发育分析。结果表明，在胁迫第12 h酶活性达到峰值，两种胁迫下野生紫花苜蓿的酶活性基本高于‘阿尔冈金’紫花苜蓿，两种紫花苜蓿PEPC蛋白均为不稳定蛋白和亲水性蛋白。干旱胁迫与高温胁迫都对紫花苜蓿PEPC酶活性产生了较大的影响，且两种紫花苜蓿PEPC氨基酸序列保守性均较高。本研究可为提高高温和干旱条件下紫花苜蓿的光合特性进而增加产量提供理论依据。     

不同紫花苜蓿品种在均匀与不均匀盐胁迫下的耐盐性评价

土壤中的盐分分布通常存在空间异质性,单株植物的根系区域土壤盐分变化较大。而目前关于植物品种耐盐性的评价都是在均匀的盐胁迫下进行的,研究不均匀盐胁迫下植物品种的耐盐性更接近自然盐分状况,可以为耐盐品种的选育提供重要的参考依据。选取6个紫花苜蓿品种作为研究对象,通过分根装置在均匀(100/100、200/200mmol·L-1 NaCl)盐胁迫和不均匀(0/200mmol·L-1 NaCl)盐胁迫下分别测定植株生长速率、地上生物量、地下生物量、水分吸收、电导率、丙二醛(MDA)、叶绿素(Chl)、脯氨酸(Pro)含量等指标,对各紫花苜蓿品种进行耐盐性比较;并通过隶属函数法对6个紫花苜蓿品种各盐分浓度下的耐盐性进行综合评价。结果表明,不论在哪种盐胁迫(均匀、不均匀)处理下,各紫花苜蓿品种均表现为生长速率下降,地上、地下生物量降低,水分吸收和叶绿素含量减少,膜透性、MDA和Pro含量增加,盐胁迫抑制了植物生长,且各指标存在品种差异。通过综合评价,均匀和不均匀盐胁迫下各品种的耐盐性不同,在均匀盐胁迫(100/100和200/200mmol·L-1 NaCl)处理下各品种的耐盐性为中苜一号WL354HQWL343HQWL353LH阿尔冈金WL298HQ,而在不均匀盐胁迫(0/200mmol·L-1 NaCl)处理下则为中苜一号WL354HQWL298HQWL343HQWL353LH阿尔冈金。中苜一号在不同盐胁迫处理下均表现出较高的耐盐性,而阿尔冈金耐盐性较差,WL298HQ在均匀与不均匀盐胁迫下的耐盐性差异较大,其他品种的耐盐性居中。     

应用隶属函数法评价10个紫花苜蓿品种的耐热性

以10个紫花苜蓿(Medicago sativa)品种作为试验材料,采用盆栽法探究38℃/35℃(昼/夜)高温胁迫对其生长和生理特性的影响,并用隶属函数法综合评价其耐热性,筛选耐热性较强及热敏感性紫花苜蓿品种.结果 表明:在高温胁迫处理下,紫花苜蓿的各项生长指标和叶绿素含量均低于对照;丙二醛(MDA)含量均大于对照;可...     

温控条件下高温胁迫对紫花苜蓿地上部分生长的影响

本研究的目的在于探讨高温对紫花苜蓿(Medicago sativa)产量和形态的影响。试验采用盆栽方法,选取6个紫花苜蓿品种,将其种植在2个生长培养箱内,2个培养箱温度(白天/黑暗)分别设置为25/20℃和35/30℃,测定其产量、株高、叶部特征及叶片叶绿素含量。结果表明,高温胁迫使紫花苜蓿生长受到抑制,植株矮小,产量降低54%;单叶片叶面积缩小至常温的1/4,但叶片厚度增加了54%。综合上述结果可以看出,高温是生产中紫花苜蓿夏季减产的一个重要原因,紫花苜蓿对高温非常敏感,但会通过一系列形态上的变化来适应高温的胁迫。     

饲料蛋白质保护对肉牛饲料氮的降解,消化和利用的影响

本研究用甲醛和新型保护剂保护豆饼日粮及未保护豆饼日粮在 3头装有瘤胃和真胃瘘管的阉牛中进行 3× 3拉丁方试验 ,研究保护及未保护豆饼蛋白质在瘤胃降解、真胃后消化吸收及吸收蛋白质的利用率的差异。结果表明 ,新型保护剂保护豆饼明显降低豆饼蛋白质在瘤胃的降解率 (p<0 .0 1 ) ,增加日粮过瘤胃蛋白质量 ,而不影响蛋白质在小肠的消化率及氮的沉积效率 ,但甲醛保护豆饼会导致吸收蛋白质的沉积效率明显下降 (p<0 .0 1 )     

生长肉鸭体蛋白周转的营养生理效应研究

选２７０只１日龄天府肉鸭，采用单因子试验设计，用同位素（Ｌ－〔４，５－＾３Ｈ〕Ｌｙｓｉｎｅ）示踪的大剂量法研究了肉鸭体蛋白周转的营养生理效应和周转代谢规律。试验结果表明：正常营养条件下，肉鸭体重、日增重、饲料利用效率与ＦＳＲ（体蛋白合成率）极显著地呈二次曲线相关（Ｒ＝１，Ｐ＜０．０１）。相对生长率、日粮粗蛋白利用效率随体蛋白周转率降低而下降（Ｐ＜０．０１）。体蛋白周转强度明显比其它禽和哺乳动物大。     

兰狐和银黑狐免疫球蛋白的提纯与鉴定

采用硫酸铵盐析和Agarose-Protein G亲和层析相结合的方法，从兰狐和银黑狐血清中提取和纯化了免疫球蛋白（IgG），应用SDS-PAGE分析表明：提纯的免疫球蛋白纯度高，重链和轻链的分子量分别为45ku和21ku左右；以犬瘟热病毒（CDV）为包被抗原的间接ELISA试验结果表明：提纯的蛋白免疫小鼠分别制备的抗兰狐和银黑狐免疫球蛋白抗体作为二抗检测CDV阳性血清具有良好的生物活性。本研究制备的高纯度的兰狐和银黑狐免疫球蛋白，对今后狐源CDV等疫病血清流行病学的调查、诊断试剂盒的开发及疫苗免疫效果的评价奠定了基础。     

Envelope protein complexes of Mycobacterium avium subsp. paratuberculosis and their antigenicity

Mycobacterium avium subsp. paratuberculosis (MAP) is the causative agent of Johne's disease, a chronic enteric disease of ruminant animals. In the present study, blue native PAGE electrophoresis and 2D SDS-PAGE were used to separate MAP envelope protein complexes, followed by mass spectrometry (MS) to identify individual proteins within the complexes. Identity of individual proteins within complexes was further confirmed by MS upon excision of spots from 2D SDS-PAGE gels. Among the seven putative membrane complexes observed, major membrane protein (MAP2121c), a key MAP antigen involved in invasion of epithelial cells, was found to form a complex with cysteine desulfurase (MAP2120c). Other complexes found included those involved in energy metabolism (succinate dehydrogenase complex) as well as a complex formed by Cfp29, a characterized T cell antigen of Mycobacterium tuberculosis. To determine antigenicity of proteins, Western blot was performed on replicate 2D SDS-PAGE gels with sera from noninfected control cows (n = 9) and naturally infected cows in the subclinical (n = 10) and clinical (n = 13) stages of infection. Clinical animals recognized MAP2121c in greater proportion than subclinical and control cows, whereas cysteine desulfurase recognition was not differentiated by infection status. To further characterize antigenicity, recombinant proteins were expressed for 10 of the proteins identified and evaluated in an interferon-gamma (IFN-γ) release assay as well as immunoblots. This study reveals the presence of protein complexes in the cell envelope of MAP, suggesting protein interactions in the envelope of this pathogen. Furthermore the identification of antigenic proteins with potential as diagnostic targets was characterized.     

成年阉牛对饲粮蛋白质的消化及维持需要量的估计

选择 6头安装有瘤胃、十二指肠前端和回肠末端瘘管的阉牛 ,分 3期依次饲喂以玉米、棉籽饼和氨化稻草为原料配合的 3种不同饲养水平的日粮 ,以Cr2 O3为指示剂测定小肠表观可消化N供给量 ,采用全收粪尿法测定氮平衡值。结果显示 ,以代谢体重为基础的氮平衡值 (NB)与进食氮和小肠表观可消化氮供给量之间分别存在如下回归关系 :NB =- 0 .5 0 12 +0 .5 76 8×进食氮　　　　　　　　R =0 .76 6 0NB =- 0 .2 80 4 +0 .6 398×小肠表观可消化氮R =0 .9894据此估计 ,阉牛维持的粗蛋白、小肠表观可消化粗蛋白和净蛋白需要量分别为 5 .4 3、2 .74、1.76g kgW0 .75。对体表氮损失加以考虑后 ,阉牛维持的粗蛋白、小肠表观可消化粗蛋白和净蛋白需要量分别为 5 .6 5、2 .93、1.88g kgW0 .75。小肠表观可消化粗蛋白用于维持和增重的效率为 0 .6 4     

鹿蛋白质营养需要研究进展

综述了近 1 0年国内外主要鹿种蛋白质营养需求的研究结果 ,分别对不同年龄、不同季节及不同生理时期鹿的蛋白质需求作了较为详细的介绍与比较 ,并提出了进一步研究方向     

Effect of a 12-Week Conditioning Period on Nitrogen Balance in Horses

Fourteen horses were used in two 12-week experiments. Horses were fed to meet crude protein needs for exercising horses and participated in light-to-moderate exercise. Total feces and urine output were measured in 4-day collections at the beginning and the end of each experiment. Fecal nitrogen (FN) and urine nitrogen (UN) were used to calculate nitrogen balance and digestibility. Nitrogen intake (NI) was higher in the end collection (P = .001). Fecal nitrogen expressed as a daily amount or as a percentage of NI was not different (P = .63 and .39, respectively). Nitrogen digestibility was not different (P = .39); however, the amount of nitrogen absorbed (NA) was lower in the initial collection (P = .001). Urine nitrogen was lower in the end collection when expressed as a total loss per day (P = .041), percent of NI (P = .001), or as a percent of NA (P = .001). Daily nitrogen retention (NR), NR expressed as a percentage of NI and as a percentage of NA, was higher in the end collection (P = .001). The improvement in NR in the end collection is due to the reduction in UN loss. This suggests more efficient use of dietary protein because there was no difference in FN loss; but without a significant change in body weight or body condition score, it cannot be concluded that the additional retained nitrogen was because of a need to support additional muscle mass.     

The response of phospholipase C/protein kinase C and adenylyl cyclase/protein kinase A pathways in porcine theca interna cells to opioid agonist FK 33-824

Opioids were found as factors affecting porcine ovarian steroidogenesis. The mechanism of opioid action, however, on porcine theca interna cells is completely unknown. Therefore, the present study was designed to investigate the possible involvement of two intracellular pathways, phospholipase C/protein kinase C and adenylyl cyclase/protein kinase A, in opioid signal transduction in porcine theca cells treated with mu opioid receptor agonist, FK 33-824. Incubation of the cells for 4 h with FK 33-824 at the dose 1 nM resulted in decreases in inositol phosphate accumulation as well as androstenedione (A(4)), testosterone (T), and estradiol (E(2)) secretions. Protein kinase C (PKC) inhibitors, staurosporine (1-100 nM), D-sphingosine (10-500 nM), and PKCi (100-2000 nM), both added alone and together with the opioid agonist, depressed release of the steroid hormones. PKC activator, phorbol ester (PMA, 1-100 nM), used alone was without effect on theca cell steroidogenesis, but added in combination with FK 33-824 abolished inhibitory influence of the opioid on A(4), T, and E(2) output. The steroid hormone secretion by PKC-deficient theca cells was inhibited by the opioid agonist. FK 33-824 also suppressed PKC activity reducing [(3)H]PDBu specific binding to theca cells, whereas ionomycin (a positive control) increased labeled phorbol ester binding to the cells. In the next experiment, cAMP release from theca cells during 2 and 4 h incubations with FK 33-824 (1-100 nM), naloxone (10 microM; opioid receptor antagonist), and LH (100 ng/mL; a positive control) was examined. FK 33-824 at the dose 1 nM inhibited cAMP secretion during 2 h incubation, but had no effect during longer incubation. LH in a manner independent on incubation time multiplied cAMP release. Protein kinase A inhibitor, PKAi (100-2000 nM), alone and in combination with FK 33-824 (1 nM), inhibited A(4), T, and E(2) secretions by theca cells. PKA activator, 8BrcAMP (10-1000 microM), stimulated the steroid hormone release, but this stimulatory effect was diminished in the presence of FK 33-824. The results allow to suggest that opioid peptides affect porcine theca cell steroidogenesis and their acute action on the cells is connected with the inhibition of phospholipase C/protein kinase C and adenylyl cyclase/protein kinase A signal transduction systems.     

Analysis of blood clotting factor activities in canine Legg-Calvé-Perthes' disease

Legg-Calve-Perthes' (LCP) disease is a noninflammatory aseptic necrosis of the femoral head and neck in small-breed dogs. The etiology of the disease is not known, but ischemia resulting from vascular compression or occlusion has been proposed. A latent ischemic phase during development of the femoral epiphysis seems to be responsible for the onset of the typical clinical features of LCP disease. Ischemia might result from insufficient oxygen supply either caused by a reduced number of afferent arterial vessels or an occlusion of the efferent venous vessels by thrombosis. In humans, LCP disease has been linked to hypercoagulability and hypofibrinolysis caused by deficiencies of protein C, protein S, or resistance to activated protein C. To determine whether canine LCP disease is caused by similar deficiencies, we determined protein C, protein S, activated protein C, factor II, factor V, factor VIII:C, and AT III activities in plasma samples of 18 dogs with clinically and histopathologically verified LCP disease. All dogs had normal plasma activities of these factors, indicating that in these dogs LCP disease was not caused by deficiencies of the analyzed blood clotting factors.     

饲粮不同蛋白质、能量水平对大白鼠生长发育及体成分的影响

本试验所用动物为53—57日龄雄性大白鼠(sprague——Dawley品系),初始体重为121.73±3.95g,试验期为大白鼠生长发育旺期的21天。试验拟研究不同粗蛋白水平(10％、15％、20％、25％和30％)及两个能量水平(消化能:3.6和3.9kcal/g)的饲粮对大白鼠生长发育及各种体成分的影响。结果表明,当饲粮蛋白质供应充分时,大白鼠生长随饲粮能量浓度的提高而改善;而当饲粮蛋白质供应感觉不足时,提高能量浓度对生长无益。机体各项成分指杯中,以蛋白质、脂肪、水分沉积受饲粮蛋白质,能量浓度影响显著;而蛋白质、脂肪沉积在很大程度上受饲粮能朊比的影响,饲粮中添加脂肪可使大白鼠机体肥胖。