火鹤花的组织培养

对火鹤花进行组培实验。1）用幼嫩的叶片和1／2MS培养基进行初代培养,继代培养培养基又恢复正常的MS,此点与以往的组培初代培养培养基的配方有区别。2）初代启动培养时间（6—8周）和继代培养（45d）周期很长与其他植物组培也有所区别。3）火鹤花因为极易生根且叶片革质不易失水,所以炼苗极其容易。实验证明,火鹤花的组培苗驯化成苗率达95％以上．使得火鹤花苗完全可以工厂化生产。     

利用腊叶标本初探椭圆叶花锚（龙胆科）花特征的地理变异

对一种植物花特征的地理变异及其选择压力研究首先需要了解该植物花特征在不同地区的变异式样,而利用腊叶标本是获取植物花特征地理变异的理想方法。我们利用椭圆叶花锚的腊叶标本,通过测量花的高度和距的长度,并与地理因子（海拔、纬度和经度）做回归分析,探讨椭圆叶花锚两个变种的花特征及其地理变异式样。结果表明：1）已鉴定为两个变种的距长度并没有明显的差异,而且变种大花花锚的距长度均在原变种卵萼花锚的变化范围之内;如果不考虑两个变种,椭圆叶花锚的距长度变化不存在明显的间断,因此研究结果不支持将椭圆叶花锚分为两个变种;2）椭圆叶花锚的花高度和距长度呈显著的正相关关系,而且99％的数据都在预测回归99％的置信区间内,表明花高度和距长度可能受到了相同选择压力的驱动;但仍有1％的点在预测区间之外,表明这些地点的花经历的选择压力可能与其它多数分布地点的选择压力不尽相同;3）椭圆叶花锚花的几何大小与纬度和海拔呈显著的负相关关系,而与经度呈显著的正相关关系;依据三个因子的回归系数,纬度（-2．735）对椭圆叶花锚花大小的影响最大,而海拔（-0．516）和经度（0．669）相对较小,因此如果要开展椭圆叶花锚花特征的地理变异研究,在纬度不同的地点开展相关研究将获得更为理想的数据。我们的研究结果表明椭圆叶花锚花特征地理变异可能存在不同的选择压力,而开展相关的野外观察和实验将极大的丰富我们对椭圆叶花锚的花特征地理变异及其选择压力的理解,并为如何利用腊叶标本开展相关研究提供一个思路。     

家蚕催青后期胚胎蛋白质双向电泳图谱分析

采用蛋白质双向电泳技术分析了家蚕Bombyx mori催青后期胚胎蛋白质图谱的变化。研究发现: 在头胸分化期（戊₃）、反转期（己₁）、毛瘤发生期（己₂）、点青期（己₃）、转青期（己₄）和孵化期（己₅）胚胎蛋白质的双向电泳图谱中共检测到209个特异蛋白斑点,其中己₃和己₄两个胚胎出现的特异蛋白斑点数在整个催青期胚胎中为最多,分别达55和77个。与催青前期胚胎出现的特异蛋白斑点变化规律相似,这些特异蛋白斑点大多也是在随后邻近的胚胎发育中消失。推测这些特异蛋白可能与相应胚胎的形体特征发育有关。     

不同生态区烟草叶片蛋白质组学的比较

为探讨不同生态区烟叶香气风格形成的机理,应用蛋白质双向电泳联用质谱技术,对河南平顶山（浓香型烟叶的典型生态区）和福建龙岩（清香型烟叶的典型生态区）的烟草（Nicotina tobaccum L．cv．K326）叶片蛋白质组成进行了比较研究。结果发现,51个蛋白质在两个生态区发生了差异表达,其中在河南表达量上升的有15个,在福建表达量上升的有25个。另外,还分别有2个和9个蛋白点为在河南和在福建样品中特异表达。采用MALDI-TOF/MS进行肽质量指纹图谱分析,经MSDB、NCBInr和SwissPort数据库查询,共鉴定出25种蛋白质,其中参与叶绿体发育、色素代谢、光合作用相关的蛋白在福建烟区高表达,与糖酵解途径相关的蛋白质在河南烟区中高表达。另外,在两生态区烟叶中都发现有相当数量的特异表达的抗逆和防御蛋白。首次在蛋白质组学水平对不同香气风格烟叶的形成机理进行了探讨。     

胃癌及癌旁组织定量比较蛋白质组学研究

为寻找胃癌特异的肿瘤标记物,用于胃癌临床诊断及药物治疗靶点的选择,本研究采用荧光差异显示凝胶电泳（DIGE）技术分离并筛选 Cy3、Cy5 及 Cy2 荧光素标记的胃癌及对应癌旁组织差异表达蛋白质,用基质辅助激光解吸电离飞行时间质谱（MALDI-TOF MS）或串联质谱技术进行鉴定并分析。结果共筛选出 33 个差异表达蛋白质点,其中 9 个蛋白质点在胃癌组织中上调,24 个蛋白质点下调。对 22 个蛋白质点采用质谱技术成功鉴定,突变结蛋白、锰超氧化物歧化酶、热休克蛋白 60等在胃癌中高表达,热休克蛋白 27、前列腺素 F 合酶、硒结合蛋白 1、锌指蛋白 160、微管蛋白 α6、真核生物翻译延伸因子 1 α1 等在胃癌组织中低表达,并筛选出 5 个未知蛋白。这些差异表达蛋白可望成为胃癌诊断的特异标记物,并与胃癌的发生、发展及预后等有关,为胃癌的诊断、发生机制的研究提供了新的思路。     

APETALA1启动子驱动AtIPT4在转基因拟南芥中表达导致花和花器官发育异常

细胞分裂素对拟南芥（Arabidopsis thaliana）花分生组织细胞的分裂和分化具有重要作用。本研究利用APETALA1（AP1）特异启动子在花分生组织和第1、2轮花器官中表达细胞分裂素合成酶（isopentyl transferase,IPT）基因IPT4,研究细胞分裂素对花和花器官发育的影响。在pAP1∷IPT4转基因植株中出现了花密集和花器官数目增多等现象。原位杂交和GUS组织染色结果发现,在pAP1∷IPT4转基因植株中,花分生组织特征决定基因LEAFY（LFY）与花器官特征决定基因AP1、PISTILLATA（PI）和AGAMOUS（AG）的表达量均有不同程度的提高。研究结果表明在拟南芥中表达pAP1∷IPT4影响其花和花器官的正常发育。     

APETALA1启动子驱动AtlPT4在转基因拟南芥中表达导致花和花器官发育异常

细胞分裂素对拟南芥（Arabidopsis thaliana）花分生组织细胞的分裂和分化具有重要作用。本研究利用APETALA1（AP1）特异启动子在花分生组织和第1、2轮花器官中表达细胞分裂素合成酶（isopentyl transferase,IPT）基因IPT4,研究细胞分裂素对花和花器官发育的影响。在pAP1∷IPT4转基因植株中出现了花密集和花器官数目增多等现象。原位杂交和GUS组织染色结果发现,在pAP1∷IPT4转基因植株中,花分生组织特征决定基因LEAFY（LFY）与花器官特征决定基因AP1、PISTILLATA（PI）和AGAMOUS（AG）的表达量均有不同程度的提高。研究结果表明在拟南芥中表达pAP1∷IPT4影响其花和花器官的正常发育。     

菠萝花发育相关基因AcMADS1的克隆与组织表达特性分析

MADS-box转录因子在植物的发育过程特别是控制花器官的诱导与发育中起关键作用。利用同源克隆结合RACE技术, 从菠萝(Ananas comosus)花中分离出1个新的菠萝MADS-box基因, 命名为AcMADS1(GenBank登录号为KC257408)。AcMADS1基因的编码区为726 bp, 编码241个氨基酸, 蛋白质分子量为27.50 kDa, 等电点为9.26。序列比对和系统进化树分析表明, AcMADS1具有保守的MADS-box及半保守的K区, 属于AGL6亚家族MADS-box蛋白。生物信息学分析表明, AcMADS1是亲水碱性蛋白, 二级结构主要以α-螺旋、无规则卷曲和折叠延伸链为蛋白质骨架, 三级结构中蛋白核心结构符合转录因子与DNA结合的常见功能域MADS-box, 而且作为转录因子定位于细胞核中。组织特异性表达分析表明, AcMADS1基因在菠萝果肉以及花器官的雌蕊、花瓣和萼片中均有表达, 但在雄蕊以及营养器官的根、茎和叶中几乎不表达; 且在花器官早期发育过程中大量表达, 后期呈下降趋势。因此推测这个基因可能在菠萝花器官发育和开花诱导过程中起重要作用。     

荒漠珍稀灌木半日花种群数量动态的谱分析

依据不同坡位的半日花（Helianthemum ordosicum）种群的野外样方调查数据,运用谱分析数学方法,对其种群数量动态变化进行了分析研究。结果表明：在以地径级差划分龄级的基础上,半日花种群天然更新过程的动态是通过半日花不同龄级的株数分布波动而表现的。不同坡位半日花种群的数量动态过程受基波影响最明显,并且波动还表现出明显的中、小周期。不同坡位种群均存在基本周期1／2的12～15mm地径增量的中等长度的周期波动,以及2．00～6．25mm地径增量的小周期波动。这种中、小周期波动与环境压力、种间竞争及自身生理特性等有关。由于不同坡位的生境条件不同,表现为从坡下部到坡上部,半日花种群数量变动的波动小周期（地径增量表型）逐渐缩短。人为干扰对不同坡位半日花种群动态的波动产生影响。半日花种群数量动态变化的这种周期性的波动可使半日花种群的自我稳定性得以维持与延续。     

家蚕催青前期胚胎蛋白质双向电泳图谱分析

为了探讨家蚕Bombyx mori胚胎蛋白质整体变化,以多化性品种P50为材料,采用蛋白质双向电泳技术及图像分析技术分析了催青前期胚胎（戊₃以前）各个时期蛋白质图谱及其变化情况。研究发现:从临界Ⅱ期（丙_２）胚胎到缩短期(戊₂)胚胎蛋白质双向电泳图谱基本稳定,存在于临界Ⅱ期胚胎的蛋白斑点在催青前期的4个胚胎中消失的个数较少,仅占22.80％,而在催青的最后2个胚胎中消失的蛋白质斑点却占48.18％;在神经沟出现（丁₁）、腹肢突起（丁₂）、上唇突起（戊₁）和缩短期（戊₂）胚胎的双向电泳图谱中能够检测到100个特异蛋白质斑点,这些特异蛋白质斑点大多在随后邻近的胚胎发育中消失,暗示了这些特异蛋白可能与相应胚胎的形体特征发育有关。     

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.     

新疆郁金香一居群个体性别7年的动态变化

在雌雄同株植物中, 花性别被认为是两性资源分配对环境条件的响应, 了解个体性别随时间的变化对探讨植物性系统的变异有重要意义。早春短命植物新疆郁金香(Tulipa sinkiangensis)多为两性花居群, 前期调查显示一些居群由雄花和两性花构成。为了探讨新疆郁金香雄花的发生与变化, 我们连续7年对一个居群的花性别及其变化动态进行了跟踪。结果显示: (1)该居群主要是由一朵花植株和两朵花植株构成, 分别占居群植株总数的74.5%和23.0%。两种性别的花随机分布在不同类型的单株上, 形成了以两性花植株为主, 含有雄株、雄花与两性花同株(andromonoecy)的居群结构。(2)雄花相对较小, 子房退化, 无胚珠发育, 在植株或居群中花期较晚出现。与相同大小的两性花相比, 雄花在单花花粉量、花粉败育率、花粉粒大小及形态、雄性功能等方面没有差异。(3) 7年间, 雄花在居群中的比例经历了2011-2014年间的显著下降(23.4-3.1%)和2015-2017年间相对稳定的零星分布(1.5-1.0%)两个阶段。居群中一朵花植株数量和两朵花植株数量在年份间呈波动性变化。(4)雄花的出现可能是植株受自身资源状况及环境条件等因素的影响在花性别选择上作出的可塑性反应。     

Proteomic Approach to Reveal the Proteins Associated with Encystment of the Ciliate Euplotes encysticus

In order to identify and reveal the proteins related to encystment of the ciliate Euplotes encysticus, we analyzed variation in the abundance of the proteins isolated from the resting cyst comparing with proteins in the vegetative cell. 2-D electrophoresis, MALDI-TOF MS techniques and Bioinformatics were used for proteome separation, quantification and identification. The comparative proteomics studies revealed 26 proteins with changes on the expression in the resting cysts, including 12 specific proteins and 14 differential proteins. 12 specific proteins and 10 out of the 14 differential proteins were selected and identified by MALDI-TOF MS. The identified specific proteins with known functions included type II cytoskeletal 1, keratin, Nop16 domain containing protein, protein arginine n-methyltransferase, epsilon-trimethyllysine hydroxylase and calpain-like protein. The identified differential proteins with known functions included Lysozyme C, keratinocyte growth factor, lysozyme homolog AT-2, formate acetyltransferase, alpha S1 casein and cold-shock protein. We discussed the functions of these proteins as well as their contribution in the process of encystment. These identified proteins covered a wide range of molecular functions, including gene regulation, RNA regulation, proteins degradation and oxidation resistance, stress response, material transport and cytoskeleton organization. Therefore, differential expression of these proteins was essential for cell morphological and physiological changes during encystment. This suggested that the peculiar proteins and differential proteins might play important roles in the process of the vegetative cells transforming into the resting cysts. These observations may be novel findings that bring new insights into the detailed mechanisms of dormancy.     

Fossil palm flowers in Dominican and Baltic amber

Five palm flowers in Dominican amber and one in Baltic amber are described or characterized. Palaeoraphe dominicana gen. et sp. nov. in the subtribe Livistoninae, is described from one perfect flower in Dominican amber. Roystonea palaea sp. nov. is described from one staminate and one pistillate flower in Dominican amber. Three other palm flowers, two perfect flowers from Dominican amber and one staminate flower from Baltic amber, are briefly characterized and figured.  © 2002 The Linnean Society of London, Botanical Journal of the Linnean Society , 2002, 139 , 361–367.     

Construction of a proteome profile and functional analysis of the proteins involved in the initiation of mouse spermatogenesis

Spermatogenesis is a complex process of terminal differentiation wherein mature sperm are produced. In the first wave of mouse spermatogenesis, different spermatogenic cells appear at specific time points, and their appearance is expected to be accompanied by changes in specific protein expression patterns. In this study, we used 2D-PAGE and MALDI-TOF/TOF technology to construct a comparative proteome profile for mouse testis at specific time points (days 0, 7, 14, 21, 28, and 60 postpartum). We identified 362 differential protein spots corresponding to 257 different proteins. Further cluster analysis revealed 6 expression patterns, and bioinformatics analysis revealed that each pattern was related to many specific cell processes. Among them, 28 novel proteins with unknown functions neither in somatic cells nor germ cells were identified, 8 of which were found to be uniquely or highly expressed in mouse testes via comparison with the GNF SymAtlas database. Further, we randomly selected 7 protein spots and the above 8 novel proteins to verify the expression pattern via Western blotting and RT-PCR, and 6 proteins with little information in testis were further investigated to explore their cellular localization during spermatogenesis by performing immunohistochemistry for the mouse testis tissue. Taken together, the above results reveal an important proteome profile that is functional during the first wave of mouse spermatogenesis, and they provide a strong basis for further research.     

鼻咽癌5-8F细胞膜蛋白质组初步分析

细胞膜蛋白是细胞的重要组成部分,作为细胞的"门铃"与"门户",参与细胞内外物质交换、信息转换、细胞生长发育、细胞迁移以及免疫应答等重要生理活动.为鉴定鼻咽癌转移相关膜蛋白,运用差速离心联合双水相方法分离纯化鼻咽癌高转移细胞5-8F的细胞膜,SDS-PAGE分离膜蛋白,液相色谱/电喷雾串联质谱分析(LC-MS/MS)结合生物信息学分析鉴定出316种非冗余蛋白质,其中152种(48.5%)被注释为膜蛋白或膜相关蛋白.通过肿瘤差异蛋白质组数据库(dbDEPD)搜索,发现在114个膜蛋白中有49种膜蛋白与其它肿瘤的发生发展密切相关,其中21个膜蛋白与肿瘤转移相关.进一步分析发现膜蛋白CD104、VDAC2、CD298和SLC25A3与同属头颈部肿瘤的口腔癌转移相关,提示这4个膜蛋白也可能是鼻咽癌潜在的转移相关蛋白.研究结果提供了一个鼻咽癌细胞5-8F包含中高丰度膜蛋白的数据库,为进一步研究头颈部肿瘤鼻咽癌癌变分子机理积累了有价值的资料.     

应用iTRAQ技术分析滩羊二毛期不同花穗型裘皮差异蛋白

为了解二毛期时滩羊串子花型裘皮与其它类型裘皮中蛋白质的差异,本试验采用iTRAQ技术及LC-MS/MS蛋白质组学研究方法,对串子花型、软大花型、绿豆丝型及其它不规则型花穗裘皮蛋白质进行鉴定和筛选,并运用Proteome Discoverer l.4软件进行定量分析,结合数据库搜索,鉴定出具有显著表达差异的蛋白,同时应用生物学技术对其进行GO和Pathway分析。结果显示4类花穗型裘皮共检测出2 886个蛋白,其中有135个、142个、113个差异蛋白分别存在于软大花型与串子花型、绿豆丝型与串子花型、其它不规则型与串子花型3个对比组中。对有表达差异的蛋白进行分析,发现膜联蛋白与血管内皮生长因子可能与软大花型毛股形成相关,KAP3和KAP6可能与滩羊串子花型毛股结构相关。研究发现滩羊不同二毛裘皮蛋白水平上的差异,可为选育优良的滩羊串子花型二毛裘皮提供理论基础。     

Proteomic Analysis of Osmotic Stress-Responsive Proteins in Sugarcane Leaves

Osmotic stress-related proteins in sugarcane were identified using proteomics approach based on two-dimensional polyacrylamide gel electrophoresis (2-DE) and matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF-MS). Sugarcane settlings were subjected to osmotic stress in the nutrient solution containing 10% (w/v) PEG 6000 for 14 h. Total proteins were extracted from leaves, and separated by 2-DE. Four typical spots exhibited significant changes in PEG treatment compared to control, which were identified using MALDI-TOF-MS successfully. The drought inducible 22 kDa protein and Rubisco small subunit were up-regulated while isoflavone reductase-like (IRLs, related to antioxidant defense system) protein and delta chain of ATP synthase were down-regulated by the osmotic stress. Analysis of the results showed that the most differential proteins under osmotic stress were acidic, unstable and transmembrane proteins, enriched with hydrophobic amino acids such as leucine and alanine which are extremely important for structural stabilization of proteins by hydrophobic interaction. However, the drought inducible 22 kDa protein was a hydrophile and non-transmembrane protein enriched with glutamic acid. These results provide new insight into the part of regulatory mechanism of adaptations to osmotic stress through differential expression of specific proteins and implicate several previously unrecognized proteins to osmotic stress.