爬行卫矛贮藏蛋白的初步研究

以不同季节的爬行卫矛枝条为研究对象,利用光学显微技术和SDS-PAGE技术来研究其休眠期和生长期枝条中贮藏蛋白的分布情况.经汞-溴酚蓝染色后的石蜡切片显微观察表明:休眠期枝条细胞的液泡中有大量蓝色小球状蛋白颗粒,有的甚至充满整个液泡,而在生长期枝条细胞中则未发现这种蛋白颗粒的存在.SDS-PAGE结果表明:与生长期枝条样品相比,有一种分子量约为45 kD蛋白质在休眠期枝条大量特异表达,这可能就是贮藏蛋白.     

浅谈园林植物的压条繁殖法

<正>压条繁殖法是利用生长在与母树上的枝条埋入土中或用其他湿润的材料包裹,促使枝条的被压部分生根,以后再与母株割离,成为独立的新植株。压条法多用于一些花灌木及一些果树的繁殖上。1压条的时期压条的时期依压条方法不同而异,可分为休眠期压条和生长期压条两类。1休眠期压条。休眠期压条一般在秋季落叶后或早春发芽前进行,常利用1~2年生的成熟枝在休眠期进行,这种压条属于真正压条法。多用于普通压条方法进行压条。2生长期压条。生长期压条在     

黄瓜幼叶细胞中钙调素的免疫胶体金定位

 采用免疫胶体金电镜技术对黄瓜幼叶叶肉细胞中的钙调素(CaM)进行定位。结果表明：适温下生长的黄瓜幼苗叶肉细胞中的CaM主要分布于细胞核和叶绿体内，细胞核中CaM主要存在于染色质和核仁上，叶绿体中CaM主要存在于类囊体膜上；线粒体中也有一定量的CaM分布；细胞质、液泡、液泡膜及质膜上只有少量CaM存在；而细胞壁和细胞间隙却很难发现显示CaM存在的金颗粒。     

核桃树修剪要适期

<正>核桃树冬季修剪极易引起伤流,使树体水分和贮藏养分大量流失,导致树势衰弱,甚至枝条枯死,严重影啊树体正常生长和结果.核桃树伤流主要发生在休眠期;果实生长期有一定伤流;芽体萌动至盛花期,果实采收至落叶期内基本没有伤流.因此,整形修剪宜选在秋季果实采收后至落叶期和春季     

生长期喷施氨基酸硒叶面肥对苹果树腐烂病的控制作用研究

在苹果树生长期连续多年喷施氨基酸硒叶面肥的基础上,通过田间调查与室内离体枝条接种,调查氨基酸硒叶面肥对苹果树腐烂病的控制作用。结果显示,生长期喷施氨基酸硒叶面肥可明显提高苹果树枝条对苹果树腐烂病的抗扩展能力,休眠期和生长期离体枝条测定病斑长度分别为15.7、9.7 mm,显著低于对照;喷施氨基酸硒叶面肥新病增长率分别为92.79%、100.98%,显著低于对照;田间平均防效分别为11.52%、27.23%,对控制苹果树腐烂病的发生有一定的作用。     

核桃枝条中总酚、黄酮类化合物含量研究

对祁县6个核桃品种1年生枝条韧皮部的总酚、黄酮类化合物含量进行了测定,并比较不同树龄间和不同品种间总酚、黄酮类化合物含量的差异。结果表明,核桃1年生枝条韧皮部总酚、黄酮类化合物含量在不同生长发育期存在显著差异,休眠期总酚、黄酮类化合物含量显著高于生长期;不同品种、不同树龄核桃1年生枝条韧皮部的总酚、黄酮类化合物含量差异显著;还对不同品种、不同树龄核桃1年生枝条韧皮部中总酚、黄酮类化合物含量与抗寒性的关系进行了分析。     

套袋对红富士苹果果皮细胞超微结构的影响

红富士苹果果皮细胞超微结构的观察结果表明,未套袋颗粒果皮表层细胞叶绿体片层结构发育较套袋果实发达,至果实采收前,未套袋果液泡染色较深县均匀,而套袋果液泡染色深,呈絮状凝结,这可能是颗粒去袋后花青苷迅速合成,在液泡内大量积累所致。     

磷饥饿番茄液泡膜蛋白组分的适应性变化研究

通过溶液培养试验,研究了磷饥饿下番茄幼苗可溶性蛋白及根部液泡膜蛋白适应性的变化.结果表明:磷饥饿番茄幼苗根中可溶性蛋白质含量显著高于对照,而茎叶中可溶性蛋白质含量变化不显著;另外,茎叶中可溶性蛋白变化比根中迟;磷胁迫番茄幼苗根中的磷可能被转移到茎叶中重新利用.磷饥饿番茄幼苗根中液泡膜蛋白含量与各自的对照大体相当;通过SDS-PAGE检测到磷饥饿番茄液泡膜蛋白中新增49.9、52.6 kD多肽谱带,由于磷饥饿处理时间相对较短,尚未对番茄幼苗的生长造成严重影响,所以,该多肽(49.9、52.6kD)可能是磷饥饿特异诱导多肽.     

查耳酮异构酶在葡萄果实发育过程中的表达及亚细胞定位研究

以‘赤霞珠’葡萄果实为材料,利用制备好的葡萄查耳酮异构酶(CHI;EC 5.5.1.6)CHI多克隆抗体,采用RT-PCR、蛋白质印记杂交和胶体金免疫电镜技术对葡萄果实发育过程中CHI基因、蛋白表达及亚细胞定位进行了研究。结果表明:CHI基因和蛋白的表达随着果实发育有着明显的变化,在果实发育早期和转色期表达量较高,同果实中总类黄酮含量的积累变化一致;胶体金免疫定位显示在果实发育早期CHI主要分布于葡萄果皮细胞的细胞质中,在液泡及质体中也有少量分布;在转色期主要定位于细胞质、质体和细胞核中,其数量明显增多。在成熟果实中,主要存在于细胞质,少量存在于质体和液泡中。     

甜瓜雄花的几个胚胎学特征

为阐明甜瓜雄花的胚胎学特征,用光学显微技术观察雄花花粉和花药壁的发育过程。结果表明,花粉母细胞减数分裂伴随胼胝质壁发育,减数分裂产生的4个小孢子在共同的胼胝质壁中呈四面体形排列。成熟期花粉粒直径约45μm,积累大量脂滴,故属于脂质花粉。花药壁由外向内依次为表皮、药室内壁、中层和绒毡层,各含1层细胞;绒毡层在原位解体,为腺质绒毡层。表皮产生单细胞表皮毛,细胞膨大、伸长,细胞质浓,不含大液泡,仅含小液泡,生存至花粉成熟时期。     

Botulinum neurotoxin type A heavy chain intervenes in expression of general proteins in spinal cord after unilateral spinal cord injury in rats

AIM: To observe the effect of botulinum neurotoxin type A heavy chain (BoNT/A HC) on the pattern of spinal protein expression by intrathecal injection after spinal cord injury in rats, and to explore the role of BoNT/A HC intervention in spinal protein expression and some of its mechanisms in nerve regeneration after injury. METHODS: The model of unilateral lumbar spinal cord injury was established. The effects of BoNT/A HC intervention at different doses (2 μg, 4 μg, 6 μg and 8 μg) on the general pattern of protein expression in the spinal cord tissues at the injury site and the cranial part adjacent to the injury site was measured and evaluated by SDS-PAGE and Coomassie brilliant blue staining first, and then by two-dimensional SDS-PAGE. RESULTS: The histological structure of the ipsilateral side of lumbar spinal cord showed obvious destruction and degradation, mainly affecting both gray and white matter of the left side of the cord. The result of SDS-PAGE with Coomassie brilliant blue staining from injured spinal cord tissue displayed that the expression of some proteins after one-time BoNT/A HC treatment appeared obviously different from that without BoNT/A HC treatment. Moreover, the pattern of the protein expression affected by BoNT/A HC was similar to that of the normal spinal cord. The more detail information from two-dimensional SDS-PAGE indicated that more than 10 proteins with different molecular weight and isoelectronic points were differentially expressed at day 2 and day 20 after local injection of 6 μg BoNT/A HC. This altered expression actually appeared a tendency toward the pattern shown in normal group. CONCLUSION: The immediate application of BoNT/A HC at the injury site after unilateral lumbar spinal cord injury is able to affect the pattern of local protein expression. The altered protein expression by injury could be reversed back to normal or approximately normal by local BoNT/A HC administration.     

Biomass yield and accumulations of bioactive compounds in red amaranth (Amaranthus tricolor L.) grown under different colored shade polyethylene in spring season

Biomass yield and accumulations of betacyanin, chlorophyll, total polyphenol, and antioxidant activity were evaluated growing red amaranth (Amaranthus tricolor L.) in spring season under five different shades made of white, blue, green, yellow, and black polyethylene, and non-shaded frame. Temperature and light intensity varied significantly (P < 0.05) under the different colored polyethylene shades, and these differences in microclimate are implicated for the variability in growth and accumulations of bioactive compounds. The highest temperature was obtained under the blue polyethylene shade and the plants achieved highest plant height, stem length and leaf number, fresh and dry matter biomass, betacyanins, total polyphenol, and antioxidant activity. The achieved biomass yield and accumulation of bioactive compounds were almost similar to field grown red amaranth in summer season. Blue polyethylene shade has probably ensured the optimum microclimate for growth and development of red amaranth in the studied growing period. Exceptionally the plants grown under green polyethylene shade accumulated highest chlorophyll. Although non-shaded plants received highest sunlight intensity but gave the poor biomass yield as well as accumulated less bioactive compounds than plants grown under blue polyethylene shade due to low air temperature. Thus, the results indicated that blue polyethylene has potentials to increase the yield with health beneficiary bioactive compounds betacyanins, polyphenol and antioxidant activity during the low temperature regime in spring season.     

苹果属小金海棠转录因子MxMYB1基因的克隆及其原核表达

 MxMYB1是苹果属小金海棠MYB类转录因子。MxMYB1含1个重复序列及MYB蛋白特有的氨基酸组成。酶切、PCR扩增及测序分析表明构建的原核表达载体结构正确, 未出现碱基突变及移码现象。1 mmol/L IPTG诱导2 h后, 在预期的蛋白分子量38 kD处出现1条表达加强的蛋白条带, 而未经诱导的转化子没有此蛋白条带。为进一步目的蛋白的纯化和鉴定提供试验基础。     

Effect of crystallin βB2 on the aging of lens

AIM: To study the effect of crystallin βB₂ on the aging of lens. METHODS: SD rats were maintained routinely and killed at 6 different ages (1 d, 8 d, 2 weeks, 8 weeks, 8 months and 1.5 years). Water-soluble crystallins were extracted and separated by two-dimensional polyacrylamide gel electrophoresis (IEF/SDS-PAGE). After Comassize blue staining, the crystallin patterns were screened and analyzed. βB₂ crystallin and the main chaperone proteins (αA₂, αB₂) were identified and the relative quantity was measured. RESULTS: (1) The quantity of water-soluble crystalline βB₂ increased in close relation to the aging of the rat. (2) αA₂, αB₂ chaperone proteins increased with the aging of the rat too. (3) The change of the quantity of water-soluble crystalline βB₂ was closely related to the changes of αA₂ , αB₂ chaperone proteins. (4) Degraded and modified crystallins began to appear clearly in the lens after 8 months old. CONCLUSION: Based on our results, we infer that water-soluble crystalline βB₂ increases with the aging dof the rat, which is helpful to maintain the structure and transparency of the lens.     

甘蓝柱头蛋白质双向电泳体系的建立

蛋白质提取方法和电泳条件是蛋白质组学研究中双向电泳的关键。为建立适合甘蓝柱头总蛋白质的双向电泳体系,本试验以甘蓝柱头为材料,采用改良TCA/丙酮沉淀法提取柱头总蛋白质,通过对IPG胶条、上样量、聚焦程序、SDS-PAGE胶浓度等的优化,建立了甘蓝柱头总蛋白质双向电泳体系,即选用长17 cm、pH 3～10 NL的IPG胶条,上样量150 μg,按照聚焦程序Ⅰ进行等电聚焦,第二向SDS-PAGE胶浓度选用12%,得到了分辨率高、重复性好的双向电泳图片。     

鹤望兰八氢番茄红素脱氢酶基因SrPDS 的克隆及表达分析

 采用RT-PCR和RACE方法从鹤望兰黄色花萼中克隆到类胡萝卜素合成途径关键基因SrPDS, 该 cDNA 全长2 069 bp,具有完整的开放阅读框（ORF）,共1 746 个碱基,编码581 个氨基酸。氨基酸 同源性分析表明,SrPDS 推导的氨基酸序列与已报道的其他植物的PDS 蛋白具有很高的同源性。系统进 化树分析显示,鹤望兰SrPDS 与番红花首先聚为一类,其次与百合、水仙PDS 蛋白亲缘关系较近。应用 半定量PCR 分析表明,SrPDS 在黄色花萼和叶片中高表达,在蓝色花瓣和根中低表达;且在花发育的始 花期表达量最高,表明该基因在黄色花萼发育过程中起着重要作用。     

基于蛋白质组学研究光质对番茄果实挥发性物质的影响机理

以‘Micro-Tom’番茄(Solanum lycopersicum L.)为试材,种子发芽后30d移入人工气候室,分别进行红/蓝光组合(1:1、3:1、5:1、7:1)处理,以白光处理为对照。在番茄果实绿熟期、转色期、成熟期取样,测定果实挥发性物质含量及蛋白组数据。结果表明,在转色期和成熟期,对番茄风味有积极作用的己醛、反式–2–己烯醛、β–紫罗兰酮、牻牛儿丙酮、6–甲基–5–庚烯–2–酮、2–苯乙醇、愈伤木酚的含量在红/蓝光3︰1组合处理中最高,而对风味有消极作用的水杨酸甲酯的含量较低,并且在成熟期为0。进一步分析了红/蓝光3︰1组合处理与对照相比的差异蛋白,在成熟过程中番茄果实共鉴定到12个与挥发性物质产生有关的差异蛋白,其中8个上调表达,4个下调表达。通过对12个差异蛋白进行mRNA水平的验证发现,只有转色期的苯丙酮酸互变异构酶(MIF)在蛋白质和mRNA水平表达不一致,其他11个蛋白在两个水平表达均一致。     

Expression of GST fusion proteins of human cytochrome P2B6 and preparation of anti-cytochrome P2B6 polyclonal antibody

AIM: To get large amounts of pure antigens to raise specific antibodies and to perform quantifications.METHODS: CYP2B6 (cytochrome P) cDNA fragments was ligated into BamHI restricted PGEX-3b to generate recombinants PGEX/2B6. We identified recombinants PGEX/2B6 by EcoRI digestion. The expression of fusion proteins were induced by adding isopropyl-thiogalactoside(IPTG). Several clones showed high-level expression of fusion proteins. Insoluble proteins was isolated from the bacteria and the fusion proteins was recovered and purified from a preparative (2mm) SDS-PAGE. The polyarcrylamide gel containing the fusion proteins glutathione S-transferase(GST-2B6) were used to immunize BALB/C mice from which polyclonal ascites fluid was prepared. The purified fusion proteins GST-1A1(GST fusion protein of CYP1A1 cDNA246~386aa expressed in this library, purified by preparative SDS-PAGE), GST-2B6 were used to test the specificity of 2B6pAb. RESULTS:Fusion proteins constructed between GST and CYP2B6 was expressed in Escherichia coli DH5α. Mouse antibodies are raised against the fusion proteins GST-2B6. 2B6pAb was fond to be specific antibody.CONCLUSION:Recombinant PGEX/2B6 were constructed and purified fusion proteins GST-2B6, and specific 2B6pAb were obtained.     

银杏叶片可溶性蛋白质测定非直线性原因分析

曾报告用考马斯亮蓝法测定银杏叶片可溶性蛋白质含量缺乏直线性,猜想其可能具有较高的蛋白水解酶活性。研究表明,银杏叶片蛋白水解酶用ｐＨ７．０缓冲液提取时活性最高,酶活最适ｐＨ５．５~６．０,因而不能解释ｐＨ８．３缓冲液提取出的蛋白质含量下降的原因。比较不同的蛋白质测定方法表明,双缩脲法测得的ＯＤ５４０值相当稳定,而考马斯亮蓝法测定的ＯＤ５９５值随时间延长而升高;同样比色５ｍｉｎ,双缩脲法测得的蛋白质浓度比考马斯亮蓝法高２０~３０倍,说明银杏叶片蛋白质不能迅速溶解于酸性考马斯亮蓝显色液是其测定非直线性的主要原因。