克鲁兹王莲(睡莲科)的开花生物学研究

分别于2003和2004年8月在北京地区观察了户外种植的克鲁兹王莲(Victoria cruziana)花的生物学.结果显示:(1)克鲁兹王莲的花在连续2 d内开放和关闭,第1天花开放的时间为19:20;第2天于12:00~13:00完成关闭过程,第2天15:00~16:00花第2次开放,第3天早晨花关闭后沉入水下.(2)开花第1天晚上花内温度高出环境温度最高达11°C,开花的第2天以后,花内温度与水表面温度相近.(3)克鲁兹王莲的花第1次和第2次开放以及关闭过程中花器官呈现有规律的运动,第1天晚上花的开放过程可划分为4个时期,花的第1次关闭与第1次开放过程中花器官的运动不是简单的相反过程,第2次开放与第1次开放的过程也不相同.(4)首次描述了拟雄蕊、雄蕊以及伴生心皮(paracarpels)在开花和关闭过程中的活动规律;开花第1天晚上的柱头是湿润的,但并不像睡莲属(Nymphaea)植物那样积累1池粘液.(5)在北京地区,克鲁兹王莲自花受精能够产生种子.研究结果认为王莲属植物的花具有很多高度特化的特征以适应昆虫传粉.     

濒危植物矮沙冬青开花物候研究

对矮沙冬青(Ammopiptanthus nanus Cheng f.)的形态学特征、开花物候及花空间分布进行了研究,结果表明:在单花、花序、单株、居群水平上的开花物候都表现出很强的开花不同步性,在单花、单花序水平主要表现在单花寿命4~9 d不等,平均(6.74±1.23)d,以7 d出现的频率最高;单花序开花持续时间范围在4~13 d,平均(8.33±2.63)d,以10 d出现的频率最高;单株及居群水平上表现更为明显.矮沙冬青的花具有虫媒传粉的特征,花主要分布在花枝的上部,下部花所占比例很小,有很多花枝花全部为上部花.相关性分析表明上部花和总花数相关性极显著(r=0.990,P<0.01),而中部花、下部花和总花数的相关性不显著,甚至呈负相关.单株不同方位开花数量差异不显著(F=0.0579,P>0.05).     

不同芸豆品种种子发育过程中贮藏蛋白积累研究

以半蔓生型奶花芸豆(Y09)和直立型奶花芸豆(Y06)为材料,通过SDS-PAGE分析研究了开花后种子形成过程中贮藏蛋白(SP)的积累规律.结果表明:2个芸豆品种开花后种子形成过程中,子叶、胚蛋白积累变化趋势基本一致,但其变化幅度存在显著差异,其中,在开花后20d内,半蔓生型芸豆Y09的籽粒蛋白积累量大于直立型芸豆Y06,但在开花后25~40d,Y06籽粒蛋白积累量显著高于Y09;两芸豆品种的子叶和胚均含有丰富的贮藏蛋白(SP)(14.4~97.4kD),其中子叶的41.0、43.9、39.4kD3种蛋白含量较高,约占子叶总蛋白含量(光密度)50%以上,而胚蛋白亚基分布相对均匀;两品种的子叶、胚中蛋白亚基差异较小,只有个别亚基存在差异.     

大豆花荚败育期间的植物激素变化

大豆开花结荚期,不同发育阶段的幼蕾与花荚的脱落率不同,其中以花后5d内的幼荚脱落最严重。与败育花荚相比,正常花荚中的干物质积累量均较高。细胞分裂素(DHZRs,ZRs,iPA)含量也较高,花后3～5d的幼荚中表现更明显。脱落酸(ABA)则是以败育幼蕾及花后3～5d的幼荚中含量较高。不同发育阶段的大豆生殖器官中,正常开放花中的玉米赤霉烯酮(ZEN)含量最高。     

科尔沁沙地两种蒿属固沙半灌木的开花物候比较研究

对科尔沁沙地的乌丹蒿和差不嘎蒿的开花物候进行了调查研究,从头状花序、花序枝、个体以及群体水平比较两种植物的开花物候特性。结果表明:(1)乌丹蒿的群体始花期为6月18日,较差不嘎蒿的早9 d,持续时间为19 d,差不嘎蒿开放持续16 d,两种植物群体花期重叠约10 d。在个体水平上,乌丹蒿始花日期比差不嘎蒿早7 d,单株花期长2 d;(2)乌丹蒿二级花序枝上头状花序数比差不嘎蒿的少,开放持续时间更长。两种植物开花频率均为单峰型,二级花序枝开放持续时间频率均以8 d为最高,其次为9 d,头状花序水平上两者开放持续时间频率较高的均为5 d、4 d、6 d,5 d频率最大;(3)差不嘎蒿二级花序枝上近基部的头状花序先开放,自基部向上的第2和第3个位置最先开放的频率相对为高,分别为12.7%和11.9%。而乌丹蒿则是顶端的头状花序先开放,最先开放的频率高达56.3%;(4)两种植物的开花振幅曲线均呈单峰曲线,其中乌丹蒿开花后第9 d开花量达到高峰,差不嘎蒿开花后第6 d开花量达到高峰。两种植物开花物候的相似性反映出它们的亲缘关系及对共同环境的适应性,不同点尤其是最先开放顺序的差异性,表明了两者长期进化的遗传差异。     

芒果APl同源基因的克隆及其生物信息学分析

我们采用RT-PCR方法克隆了2个APl同源基因全长cDNA,分别命名为MAPl-1(GenBank accession No.FJ529206)和MAPl-2(GenBank accession No.FJ529207).MAPl-1编码247个氨基酸,开放阅读框长度为741 bp,蛋白质分子量为28.54kD,等电点为8.31;MAPl-2编码248个氨基酸,开放阅读框长度为744 bp,蛋白质分子量为28.78 kD,等电点为8.70.同源性分析表明,它们的核苷酸序列与其它木本植物APl同源基因的一致性为72%～81%.实验分析表明,MAPl-1和MAPl-2第1至第61个氨基酸含有一个MADS盒结构域,第88至第178个为K盒结构域;两个基因均定位于细胞核,且功能位点分布存在着不同,推测这两个基因在花器官发育过程中的功能存在差异.蛋白二级结构预测显示,MAPl-1蛋白有12个a-螺旋,4个β折叠区,14个β-转角;而MAPl-2蛋白有11个a-螺旋,5个β折叠区,15个β-转角:其大多数氨基酸具有亲水性.本研究有助于进一步了解芒果的开花分子机理及成花的生物学发育阶段.     

迁地保护条件下两种沙冬青的开花物候比较研究

沙冬青属(Ammopiptanthus)植物是我国西北荒漠区唯一的常绿阔叶灌木。作者对吐鲁番沙漠植物园迁地保护的两种沙冬青的开花物候进行了详细的比较观察,旨在探讨它们在同一生境条件下开花特性的异同点及其影响因素。主要结果如下:(1)两种植物在开花频率、花序开放顺序、开花振幅曲线及单花寿命等开花参数上相似,但在始花时间、单株花期、花序的开花数及开放持续时间与频率分布、开花振幅等参数上明显不同;(2)在个体和群体水平上,蒙古沙冬青(A.mongolicus)始花时间均比新疆沙冬青(A.nanus)早,蒙古沙冬青开花全过程为20–21d,新疆沙冬青为13–14d;(3)蒙古沙冬青花序的开花数比新疆沙冬青多、开放持续时间长,两者在开花数(F=17.51,P<0.01)和持续时间(F=14.08,P<0.01)上均存在显著差异;(4)花序上的花大多从近基部向两端开放,开花振幅呈单峰曲线,但新疆沙冬青的开花振幅较高;(5)花序开放持续期的频率分布明显不同,新疆沙冬青较蒙古沙冬青更为集中,但两者的单花寿命稳定,均在7d左右;(6)花序上每天的开花数与其座果数呈正相关(蒙古沙冬青,r=0.885,P<0.05;新疆沙冬青,r=0.827,P<0.01),但其开花数和座果数与始花时间存在不同程度的相关关系,这些特点可能与开花对传粉者的吸引以及物种本身的遗传特性有关。对上述观察结果及其影响因素的分析表明,两种植物在开花参数上所表现出的一致性可能是受系统发育限制的,而彼此间的差异可能与其进化历史及所处的环境异质性有关,是在与环境的长期适应过程中分别形成的一些可遗传的变异;而不同年份间两种沙冬青在花序的开花数及开放持续时间上表现出的差异可能与环境温度的变化有关。这些结果对于探讨该属植物的繁殖生物学特性及其保护对策具有重要意义。     

广州地区3种睡莲花开放的日变化规律分析

为了解睡莲属(Nymphaea)植物开花的日变化规律,以广州华南农业大学湿地公园的睡莲属植物为对象进行观察和分析。结果显示,晚上开花类的热带睡莲,其日变化是花朵于晚上开放并持续至第二天上午,白天开花类的热带睡莲和耐寒睡莲,其开放时间为白昼,开放时间因种类不同而有所差异。白天开花的热带睡莲‘独立’(Nymphaea‘Independence’)和耐寒睡莲‘科罗拉多’(Nymphaea‘Colorado’)单朵花期3~4 d,耐寒睡莲墨西哥黄睡莲(N.mexicana)单朵花期2 d,雌雄蕊先后成熟以达到异花授粉的目的,单朵睡莲开放期间雄蕊群呈现不同形态。‘独立’睡莲呈现日出而开,日落则合的开花生物钟,10月至12月,光照时间缩短导致‘独立’睡莲开闭节律缩短。观察结果可为园林水景中睡莲属植物配置提供准确的数据支持。     

高温胁迫对黄瓜授粉后雌花中Ca2+分布、ABA及蛋白质合成的影响

用焦锑酸钾沉淀法对45℃和28℃下黄瓜(新泰密刺)授粉后柱头及子房中C a2 分布进行了电镜观察,并采用酶联免疫吸附测定法(EL ISA)测定了叶片中的ABA含量及用等电聚焦聚丙烯酰胺双向电泳(IEF-SDS PAGE)方法对其蛋白合成的变化进行了研究.结果表明,28℃时柱头中的C a2 主要分布在细胞间隙中,经45℃处理后细胞间隙和胞内C a2 水平均显著升高,胞内外C a2 浓度梯度逐步丧失;45℃处理1 h后柱头和子房中内源ABA含量显著提高;高温处理后柱头和子房中产生了一些新的蛋白质,如柱头中的(MW26.0 kD,P I 5.4)、(MW90.0 kD,P I5.1)、(MW54.0 kD,P I 4.6)、(MW41.0 kD,P I 6.2),子房中的(MW90.0 kD,P I5.2)、(MW61.0 kD,P I 5.4)、(MW48.0 kD,P I 6.4)、(MW40.5 kD,P I 4.9);另有一些蛋白质与常温相比表达量大幅上调,如柱头中的(MW67.0kD,P I 5.8)、(MW56.5 kD,P I 5.8),子房中的(MW70.0 kD,P I 5.7)、(MW57.0 kD,P I 5.7).上述结果表明,C a2 和ABA信号系统均参与了授粉后黄瓜雌性器官对高温胁迫的反应调节并最终导致基因表达发生变化.     

乌龙岭’龙眼胚胎发育时期特异性蛋白质的变化

应用IEF-SDS-PAGE技术分析龙眼胚胎分化发育过程中蛋白质组分的变化。结果表明,在各发育阶段大多数蛋白质组分的电泳图谱基本一致,但也有变化。其中花后38d存在TE1(27．1kD、p,7．3),TE2(17．5kD、pI8．2)2个特异蛋白,45d存在TE3(11．4kD、pI7．6),TE4(13．2kD、pI9．9)2个特异蛋白,52d存在TE5(22．6kD、pI7．2),TE6(18．6kD、pI8．3),TE,(23．5kD、pI3．6)3个特异蛋白。31d胚胎电泳图谱中的蛋白质点数相对较多,表明此时蛋白质旺盛合成与积累,这与蛋白含量的变化基本一致。龙眼胚胎发育过程中特异蛋白的出现或消失．对胚胎的分化发育具有重要作用。     

Mechanism of the antimutagenic effect of interferon in human fibroblasts based on induced protein analysis]

Proteins induced in human fibroblasts after treatment of some antimutagens (interferon, p-aminobenzoic acid, heating and vaccinia virus infection) were identified by means of polyacrylamide gel electrophoresis (10-15%) followed by fluorography of the gel. Influenza virus proteins (A/WSN/33) were used as markers to determine the molecular weights of the new proteins. The results obtained suggest that interferon, p-aminobenzoic acid, heating and vaccinia virus infection induced proteins with mol. weights of 24 and 18 kD except the protein with 76 kD observed only after heating insult.     

Toxin-binding proteins from midgut epithelium membranes of Anopheles stephensi larvae

Proteins of 65 and 57 kD were isolated from the apical membranes of midgut epithelium of Anopheles stephensi larvae by affinity chromatography. These proteins can specifically bind endotoxin Cry11A and activate toxin Cry4B (Cry4B-tox) under conditions of ligand blotting, and both Cry proteins compete for this binding. At least in the case of Cry4B-tox, the binding with 65 and 57 kD proteins is reversible. The ability of the products of limited proteolysis of Cry11A and Cry4B to bind the 65 and 57 kD proteins correlates with their toxicity to A. stephensi larva. The N-terminal amino acid sequence of the 57 kD protein is unique and absent in the NCBI GenBank. The proteins of 65 and 57 kD share most of the properties studied with Aedes aegypti toxin-binding proteins. It is possible that they altogether represent a novel class (or classes) of delta-endotoxin receptors.     

Membrane proteins of Aedes aegypti larvae bind toxins Cry4B and Cry11A of Bacillus thuringiensis ssp. israelensis

Proteins of molecular weight 65 and 62 kD and having affinity for toxins Cry4B and Cry11A produced by Bacillus thuringiensis ssp. israelensis have been isolated from brush border membranes of Aedes aegypti larvae using affinity chromatography. Using a ligand blotting technique, we show that the binding of these proteins to the biotinylated toxins is reversible and that the two toxins compete for binding to the two proteins. These proteins are likely to be Cry4B and Cry11A toxin receptors in gut epithelial cells of Aedes aegypti larvae.     

Maturation proteins associated with desiccation tolerance in soybean

A set of proteins that accumulates late in embryogenesis (Lea proteins) has been hypothesized to have a role in protecting the mature seed against desiccation damage. A possible correlation between their presence and the desiccation tolerant state in soybean seeds (Glycine max L. Chippewa) was tested. Proteins that showed the same temporal pattern of expression as that reported for Lea proteins were identified in the axes of soybean. They were distinct from the known storage proteins and were resistant to heat coagulation. The level of these “maturation” proteins was closely correlated with desiccation tolerance both in the naturally developing and in the germinating seed: increasing at 44 days after flowering, when desiccation tolerance was achieved, and decreasing after 18 hours of imbibition, when desiccation tolerance was lost. During imbibition, 100 micromolar abscisic acid or Polyethylene glycol-6000 (−0.6 megapascals) delayed disappearance of the maturation proteins, loss of desiccation tolerance, and germination. During maturation, desiccation tolerance was prematurely induced when excised seeds were dried slowly but not when seeds were held for an equivalent time at high relative humidity. In contrast, maturation proteins were induced under both conditions. We conclude that maturation proteins may contribute to desiccation tolerance of soybean seeds, though they may not be sufficient to induce tolerance by themselves.     

小麦幼芽水分胁迫诱导蛋白的特征

水分胁迫（－1．2MPaPEG-6000）处理萌动的小麦种子,24h后诱导小麦幼芽产生41．5kD蛋白,其含量随着胁迫时间延长明显增加,48h时含量最高,到72h后不再变化。复水后,该蛋白消失;再胁迫48h时则又出现,其含量与处理24h时相当。41．5kD诱导蛋白主要位于细胞器膜上,细胞质中几乎不存在。41．5kD蛋白主要溶于10％NaCl提取液中,其等电点为pl5．65。该蛋白的氨基酸组成中,脯氨酸含量最高,其次为丙氨酸、天冬氨酸、谷氨酸、甘氨酸,没有发现半胱氨酸和组氨酸。     

果梅完全花与不完全花的差异蛋白分析

应用双向电泳技术对果梅完全花与不完全花的蛋白质组分进行了比较分析。经专业分析软件（PDQuest）对电泳图谱分析表明两者的蛋白分布相似,在完全花中发现了1个特异蛋白、1个上调蛋白、21个下调蛋白,在不完全花中发现2个特异蛋白,这些蛋白差异点可能与雌蕊的败育有关。应用质谱技术对3个特异点及5个差异大的蛋白点进行分析,得到的肽段数据与蛋白质数据库比对发现其中一个蛋白（28．2kD,pI4．53）与光敏色素B有关。     

Identification of leukocyte cationic proteins that interact with ceruloplasmin

Proteins from leukocytes were investigated for their ability to interact with ceruloplasmin (Cp), a copper-containing glycoprotein of human plasma. Extract from leukocytes was subjected to affinity chromatography on Cp-Sepharose, after which proteins were eluted from the resin with 0.5 M NaCl in Tris-HCl, pH 7.4. SDS-PAGE of the eluate revealed protein bands with molecular weights 78, 57, 40, 30, 16, and 12 kD. Among these, Western blotting detected myeloperoxidase (57, 40, and 12 kD) and lactoferrin (78 kD). Also, the 30-kD component had a sequence (1)I-(2)I/V-(3)G-(4)G-(5)R/H at the N-terminus that is likely to indicate the presence of neutrophilic elastase, cathepsin G, proteinase 3, and azurocidin (CAP 37) - all from the family of serprocidins. Mass spectrometry of tryptic fragments indicated the presence of the 16-kD eosinophilic cationic protein (seven peptides), 27-kD cathepsin G (eleven peptides), 27-kD azurocidin (eight peptides), 29-kD neutrophilic elastase (seven peptides), and 27-kD proteinase 3 (six peptides). Myeloperoxidase was represented by 57-, 40-, and 12-kD fragments (thirteen, ten, and four peptides, respectively). Thus, interaction with Cp of five cationic proteins, i.e. of eosinophilic cationic protein, cathepsin G, neutrophilic elastase, proteinase 3, and azurocidin is reported for the first time.     

Comparative proteomics of Cannabis sativa plant tissues.

Comparative proteomics of leaves, flowers, and glands of Cannabis sativa have been used to identify specific tissue-expressed proteins. These tissues have significantly different levels of cannabinoids. Cannabinoids accumulate primarily in the glands but can also be found in flowers and leaves. Proteins extracted from glands, flowers, and leaves were separated using two-dimensional gel electrophoresis. Over 800 protein spots were reproducibly resolved in the two-dimensional gels from leaves and flowers. The patterns of the gels were different and little correlation among the proteins could be observed. Some proteins that were only expressed in flowers were chosen for identification by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and peptide mass fingerprint database searching. Flower and gland proteomes were also compared, with the finding that less then half of the proteins expressed in flowers were also expressed in glands. Some selected gland protein spots were identified: F1D9.26-unknown prot. (Arabidopsis thaliana), phospholipase D beta 1 isoform 1a (Gossypium hirsutum), and PG1 (Hordeum vulgare). Western blotting was employed to identify a polyketide synthase, an enzyme believed to be involved in cannabinoid biosynthesis, resulting in detection of a single protein.     

Identification,purification, and characterization of pathogenesis-related proteins from virus-infected Samsun NN tobacco leaves

Ten pH-3 soluble, low-molecular-weight pathogenesis-related proteins (PRs) were found to accumulate in leaves of tobacco cv. Samsun NN reacting hypersensitively to tobacco mosaic virus. Besides the previously characterized PRs 1a, 1b, 1c and 2, these proteins were provisionally designated N, O, P, Q, R, and S in order of decreasing electrophoretic mobility in native polyacrylamide gels. Two-dimensional gel electrophoresis indicated that the PRs consist of single polypeptides, except for R, which is composed of two components with slightly different molecular weights. Estimated molecular weights in SDS-containing gels were: PRs 1a and 1b 17 kD, 1c 16.5 kD, 2 31 kD, N 33 kD, O 35 kD, P 27 kD, Q 28 kD, R 13 and 15 kD, and S 25 kD. However, based on their elution from gel filtration columns and relative moblities in native gels of different acrylamide concentrations, P and Q appeared to have molecular weights similar to those of the PR 1 group. Upon chromatofocusing no additional components were resolved. The PRs were eluted between pH 7 and 4; except for R, their pIs, as judged from isoelectric focusing, appeared to lie in the range from pH 4 to 5.2. In the presence of 6 M urea PR 1a was split into two components, one of which was strongly retarded on gels, as were P and Q. None of the PRs was detected when gels were stained for glycoproteins.By combinations of gel filtration, DEAE-cellulose chromatography, and chromatofocusing, PRs 1a, 1b, 1c, 2 and N were purified, their amino acid compositions determined, and antisera raised against each of these components. By Western blotting, antisera against either PR 1a, 1b, or 1c reacted with each of the components of the PR 1 group, as well as with PR S. Similarly, the antisera against either PR 2 or N reacted with both 2 and N, as well as with O and R. On the basis of major similarities in molecular weight characteristics, amino acid compositions, and serological relationships, it is proposed to classify tobacco PRs into five groups: 1: PRs 1a, 1b, and 1c; 2: 2a (formerly 2), 2b (N), and 2c (O); 3: 3a (P), and 3b (Q); 4: 4a and 4b (the two components of R); and 5: PR 5 (S).     

Some epidemiological aspects of post-bloom fruit drop disease (Colletotrichum gloeosporioides) in citrus

Fluctuations in the incidence and amount of post-bloom fruit drop disease of citrus caused by Colletotrichum gloeosporioides in Belize prevent economic disease control. During the cooler drier months of the year when blossom infection is common there are variations in the incidence of rainfall and associated climatic parameters, and in the pattern of flowering. Large amounts of disease develop when periods of rain followed by prolonged wetness occur during peak blossoming periods. Blossoms are most susceptible during the open flower stage and infection of terminal flowers invariably results in infection of all other flowers on the spike. Disease incidence is greater in the lower parts of the trees, but flowering is greater in the upper regions. Large numbers of Colletotrichum spores are produced during wet conditions from apparently healthy leaves and from diseased flowers, but these rapidly lose viability when dried. Few spores are produced from old persistent calices (buttons). Although spores from leaves were a less potent inoculum source than those from flowers, they could provide the initial inoculum to commence flower infection when blossoming starts.