羊草没地理种群的同工酶研究

对我国从内蒙古到吉林的荒漠草原、典型草原、草甸草原、盐碱地草原的５个样点羊草的６种同工酶进行了研究,通过聚类分析发现羊草种群随着地理生态环境的不同,在同工酶水平上发生着不同程度的种内分化,羊草在同工酶水平上的分化受多个环境因子的综合影响,而且与间草耐寒性能存在的一定的内在联系。     

不同地理种群羊草的遗传分化研究

以不同种群羊草为实验材料（包括内蒙古高原的稆草原类型：荒漠草原,典型草原,草甸草原及吉林盐碱地羊草）,分别从个体（叶长,叶宽,株高,分蘖数,根茎芽数等特征及根茎叶的扫描电镜结构）,生理（在ＰＥＧ胁迫下叶片的电阻电导,ＳＯＤ和ＰＯＤ的活性,脯氨酸和丙二醛的含量等）,细胞（染色体核型）,同工酶及ＤＮＡ（ＲＡＰＤ）等几个水平较为系统地研究其遗传分化以及对水因子的适应机理,并进行了大量的聚类分析,发现：不     

松嫩平原中部地区羊草种群的遗传结构研究

松嫩平原中间地区的羊草,依其生长的微环境不同可分成灰绿色和黄绿色两种叶色类型的种群。移栽实验表明,这两类种群实际上是灰色和灰绿色两种类型的混生群体。采用淀粉凝胶电泳技术及等位酶分析方法测定了不同生境、不同分类种群的遗传结构,发现羊草种群遗传变异度很高;但是无论如何归属,这一草原区域上的羊草种群遗传分化程度很低（ＦＳＴ≤０．０３４）。这可能与羊草与异花授粉、不同类型混生及种群间基因流强度大有关。     

羊草种群克隆多样性的初步研究

 采用水平淀粉凝胶电泳技术检测了松嫩草原上两种叶色类型(灰绿型和黄绿型)羊草(Leymus chinensis)种群的克隆多样性。羊草种群中克隆多样性较高,Simpson指数平均为0.983,97.7% 的基因型为局部分布;在克隆特性上,灰绿型和黄绿型羊草也明显不同,黄绿型种群的克隆多样性低于灰绿型。由基株数据计算的种群遗传多样性与由分株计算的结果一致,进一步确认了黄绿型和灰绿型羊草间存在着明显的遗传分化。种群中不时有实生苗补充及环境异质性引起的分化选择作用可能是维持种群内基因型多样性的重要因素。     

羊草种群遗传分化的RAPD分析I：扩增片段频率的变化

松嫩草原上分布着灰绿型和黄绿型两种叶色类型的羊草,为进一步明确这两种叶色类型种群分化的分子基础 ,对其9个种群105个个体进行了RAPD分析。15个随机引物（10nt)共扩增出149个多态性片段,总的多态位点为96．1％,黄绿型与灰绿型种群间在10．1％的多态性片段上明显不同,相似生境或同一地域的种群在一些位点上表现出相似或相同的变化;9个种群都检测到了特异性扩增。     

松嫩平原中部地区羊草种群的遗传结构研究

松嫩平原中部地区的羊草,依其生长的微环境不同可分成灰绿色和黄绿色两种叶色类型的种群。移栽实验表明,这两类种群实际上是灰色和灰绿色两种类型的混生群体。采用淀粉凝胶电泳技术及等位酶分析方法测定了不同生境、不同分类种群的遗传结构,发现羊草种群遗传变异度很高;但是无论如何归属,这一草原区域上的羊草种群遗传分化程度很低(FST ≤ 0.034)。这可能与羊草为异花授粉、不同类型混生及种群间基因流强度大有关。     

典型草原建群种羊草对氮磷添加的生理生态响应

由于人类活动和气候变化的共同作用, 大气氮(N)沉降日益加剧, 使得陆地生态系统中的可利用性N显著增加, 生态系统更易受其他元素如磷(P)的限制。然而, 目前关于N、P养分添加对草原生态系统不同组织水平的影响研究较少, 相关机制尚不清楚。该文以内蒙古典型羊草(Leymus chinensis)草原为研究对象, 通过连续两年(2011-2012年)的N和P养分添加实验, 研究建群种羊草的生理生态性状、种群生物量和群落初级生产力对N、P添加的响应及其适应机制。结果表明: 羊草草原不同组织水平对N、P添加的响应不同。群落水平上, 地上净初级生产力在不同降水年份均受N和P元素的共同限制, N、P共同添加显著提高了地上净初级生产力; 物种水平上, N、P添加对羊草种群生物量和密度, 以及相对生物量均没有显著影响, 表明羊草能够维持种群的相对稳定; 个体水平上, 在正常降水年份(2011年), 羊草生长主要受N素限制, 而在湿润年份(2012年), 降水增加使得羊草生长没有受到明显的养分限制。羊草通过增加比叶面积、叶片大小和叶片N含量, 提高整体光合能力, 以促进个体生长。总之, 内蒙古典型草原群落净初级生产力受N、P元素共同限制, 作为建群种的羊草, 其对N、P添加的响应因组织水平而异, 也受年际间降水变化的影响。     

草原区不同植物群落蒸发蒸腾的研究

 在中国科学院内蒙古草原生态系统定位研究站,采用“土柱称重法”对典型草原群落蒸发蒸腾进行实验观测,主要研究结果如下：1．草原沙地、羊草草原和河漫滩草甸是本地区差异明显的三种群落。草甸蒸腾最大(9.2 mm·d-1),比另两种群落高2—3倍;草甸蒸发最小(0.4mm·d-1),约为沙地的1／4,草原 的l／8。2．羊草草原在本区分布广,大针茅草原是本区半干旱地带性代表。由于这两种群落主要有关背景值的不同,羊草群落蒸散值(4.4—5.Omm·d-1)明显高于大针茅群落(3.5—3.8 mm·d-1),而且羊草群落的T／E1)值(～2)也明显大于大针茅(～1)。在生长季节中,羊草草原蒸散随气温升高而升高,在盛夏达最高值,然后随气温降低而降低。然而由于降雨变化的影响,羊草草原蒸发蒸腾的季节变化时而出现较大幅度的波动。3．根据1989年实测的降水和羊草群落蒸发蒸腾的季节变化,分析羊草群落的水分收支,可以看到实验区羊草草原的水分收支是大体平衡的。     

火烧对羊草草原植物群落组成的影响

在羊草草原正常能着火季节内,通过人为点烧的方法,对比研究了不同时间火烧对植物群落组成的影响．结果表明,羊草－杂类草草原早春火烧后,群落密度、种类丰富度和多样性提高,均匀性降低;羊草典型草原秋季火烧后,群落密度、多样性和均匀性降低,种类丰富度提高．连续２次火烧除羊草和几种１—２年生植物密度增加外,其它各种群密度降低,群落多样性也降低,一些种类退出群落．     

羊草种群遗传分化的RAPD分析Ⅱ.RAPD数据的统计分析

对松嫩草原上分布的灰绿型和黄绿型羊草9个种群进行了15个引物的RAPD分析,统计结果表明,两类种群的扩增片段数和多态位点比率明显不同,黄绿型种群低于灰绿型,其值分别＜90与＞100,＜50％与＞70％,比较了7种不同统计方法据RAPD表型或基因型频率估算的种群遗传多样性,几种统计结果都揭示,黄绿型种群低于灰绿型种群,用F1s值矫正种群对Hardy-Weinberg平衡的偏离后,估算等位基因频率,通过Shannon指数和Nei指数估计羊草种群间分化分别为37．6％和35．7％,高于等位酶的分析,讨论比较了等位酶和RAPD分析结果的异同。     

中国石龙子不同组织三种同工酶的研究

采用聚丙烯酰胺凝胶电泳方法,研究了分布在广东韶关地区的中国石龙子（Eumeces chinensis）的肝脏、肌肉、心脏、脑、生殖腺5种组织的酯酶（EST）、淀粉酶（AMY）和过氧化氢酶（CAT）3种同工酶,旨在为华南地区中国石龙子的系统分类提供理论依据,结果表明不同的同工酶存在组织特异性,并且分布特征与其生理功能有一定关系.     

Purification and characterization of rhizopuspepsin isozymes from a liquid culture of Rhizopus chinensis

1. Rhizopuspepsin has been purified from liquid cultures of Rhizopus chinensis. 2. Purification by ammonium sulfate precipitation, affinity chromatography on pepstatin Sepharose and low/high resolution isoelectric focusing produced five isoelectric forms. 3. The two major isozymes pI 5.1 and 5.8 did not differ significantly in amino acid composition, molecular weight and enzyme activity. 4. Three minor isozymes were partially purified as pI 7.35, 7.41 and 7.9.     

Determination of the amino acid sequences of the two major isozymes of rhizopuspepsin

The amino acid sequences of the two major isozymes of rhizopuspepsin, an aspartic proteinase from Rhizopus chinensis, were determined by analyzing the tryptic peptides derived from the reduced and carboxymethylated (RCm-) derivative of each isozyme. Amino acid substitutions were shown to occur at eight positions. Rhizopuspepsin I, with an isoelectric point of 5.1, had Ile-15, Asn-61, Ser-116, Lys-162, Ile-230, Tyr-241, Asp-293, and Glu-325, whereas rhizopuspepsin II, with an isoelectric point of 5.8, had Val-15, Lys-61, Asn-116, Ser-162, Val-230, Ser-241, Asn-293, and Gln-325, the other parts of the two isozymes being identical with each other. Thus, rhizopuspepsin I had two more net negative charges than rhizopuspepsin II. This is consistent with the difference in isoelectric point of these two isozymes.     

中国石龙子的同工酶

通过聚丙烯酰胺凝胶电泳方法,对浙江温州和福建宁德中国石龙子（Eumeces chinensis)两个地理种群的皮肤、肌肉、胃、小肠、肝脏、肺、肾、心脏、性腺等9种组织中的乳酸脱氢酶、过氧化物酶和醇脱氢酶同工酶进行了研究。结果表明,中国石龙子两种群的同工酶电泳结果具有明显的地理种群特异性,且9种不同组织中的同工酶谱亦表现出组织特异性。     

鸢尾体细胞无性系的建立与变异

本文以德鸢尾,马蔺、拟鸢尾和鸢等几种宿根鸢尾为试验材料,通过花器培养建立了体细胞无性系,在多次继代培养过程中,研究了离体培养对鸢尾体细胞无性系变异的影响,并运用聚丙烯酰胺凝胶垂直板电泳技术分析了试管苗叶片的过氧化物酶同工酶。结果表明离体培养已经改变了鸢尾的遗传基础,但在形态特征、生态习性及观赏性状等方面未发生明显的表型变异。     

Human red cell acid phosphatase: purification and properties of the A, B and C isozymes

Human red cell acid phosphatase isozymes encoded by three alleles (ACP1*A, ACPI*B and ACP1*C), each of which generates two isozymes, (f) and (s), were purified to homogeneity. The molecular mass of the six isozymes (Af, As, Bf, Bs, Cf and Cs) was estimated to be 17-18 kDa, the mass of the f isozymes probably being slightly higher than that of the s isozymes. It was indicated that the isozymes react with p-nitrophenyl phosphate in the mono anionic state, and that a group with a pKa value of about 6, which may be histidine, is of importance for the catalytic function of the s isozymes. Significant differences between the f and s isozymes were observed with respect to specific activity. Km (p-nitrophenyl phosphate), Ki (p-aminobenzylphosphonic acid), amino acid composition, stability in the presence of urea, thermal stability, retention time in size-exclusion chromatography of the native isozymes and migration in sodium dodecyl sulphate polyacrylamide gel electrophoresis, In contrast, identical or similar properties were observed for the three genetically different f isozymes, and the same was the case for the three s isozymes. It is suggested that the f and s isozymes serve different functions in the cell.     

Expression analysis of polyphenol oxidase isozymes by active staining method and tissue browning of head lettuce (Lactuca sativa L.)

Browning of plant tissue is generally considered attributable to enzymatic oxidation by polyphenol oxidase (PPO). Electrophoresis followed by activity staining has been used as an effective procedure to visually detect and isolate isozymes; however, it has not been applied for examination of various PPO isozymes in lettuce. Our study demonstrated that different lettuce PPO isozymes could be detected at different pH in active staining, and multiple isozymes were detected only under alkaline conditions. As a result, we concluded that activity staining with approximately pH 8 enabled to detect various PPO isozymes in lettuce. By expression analysis of the PPO isozymes after wounding, PPO isozymes that correlated with time-course of tissue browning were detected. The wound-induced PPO may play a key role in enzymatic browning.     

Molecular basis for the isozymes of bovine glucose-6-phosphate isomerase

Glucose-6-phosphate isomerase exists as multiple, catalytically active isozymes which can be resolved by polyacrylamide gel electrophoresis, isoelectric focusing, and ion-exchange chromatography. GPI from bovine heart was purified to homogeneity and each of the isozymes resolved. Four of the five isozymes were characterized with regard to their physical, chemical, and catalytic properties in order to establish their possible physiological significance and to ascertain their molecular basis. The isozymes exhibited identical native (118,000) and subunit (59,000) molecular weights but had different apparent pI values of 7.2, 7.0, 6.8, and 6.6. Kinetic constants, such as turnover number, Km and Ki values, were identical for all isozymes in either reaction direction. Structural analyses showed that the amino termini were blocked and the carboxyl terminal sequences were -Glu-Ala-Ser-Gly for all four isozymes. The most basic isozyme was more stable than the more acidic isozymes at pH extremes, at high ionic strength, in the presence of denaturants, or upon exposure to proteases. When the most basic isozyme was incubated in vitro under mild alkaline conditions, there was a spontaneous generation of the more acidic isozymes with electrophoretic properties identical to those found in vivo. The simultaneous release of ammonia along with the spontaneous shift to more acidic isozymes indicates deamidation as the molecular basis for the formation of the acidic isozymes both in vivo and in vitro. The change in the peptide fragmentation patterns following cleavage by hydroxylamine further suggests that deamidation of specific Asn-Gly bonds accounts for the structural basis of the isozymes.     

社鼠组织器官同工酶的研究

用聚丙烯酰胺凝胶等电聚焦电泳方法,分析了社鼠的肝、肾、心肌、骨骼肌、肺、脾和脑等多种组织器官的ＬＤＨ、ＡＤＨ、ＥＳＴ、和ＳＯＤ４种同工酶,对各组织器官的酶带数目和分布,以及酶活性进行了比较研究。结果表明,社鼠的ＬＤＨ同工酶、ＡＤＨ同工酶和ＥＳＴ同工酶具有比较明显的组织特异性,而ＳＯＤ同工酶的组织特异性较低。肺和脾除ＥＳＴ同工酶活性较高外,脑除ＬＤＨ同工酶活性较高外,其它３种同工酶的活性均较低;而肝和肾中４种同工酶的活性普遍很高。心肌和骨骼肌因氧张力不同而使ＬＤＨ同工酶酶谱存在明显差异,但其它３种同工酶酶谱却非常相似。同工酶的组织特异性与各组织器官所执行的生理功能是相一致的