小麦杂交坏死蛋白质组学研究

小麦杂交坏死是某些小麦杂交种表现出的叶片提前逐渐死亡的现象。它是由两个坏死基因Ne1和Ne2在杂交种中相遇后发生显性互补引起的。坏死从叶片尖端逐渐过渡到叶片基部，从成熟叶片发展到幼嫩叶片。一些严重坏死的F1完成它的生活周期前就在不同的生长阶段死去，无法获得F1种子，这就限制了携带优良性状的亲本的选择和优良基因的交流。另外，小麦杂交坏死是一个独特的研究植物程序性死亡的遗传系统。虽然小麦杂交坏死这种现象已经发现很多年，但其详细的分子机理却仍然未知。对小麦杂交坏死的分子机理进行深入研究将有助于克服小麦杂交利用中杂交坏死的遗传障碍，此外，也为深入研究植物的PCD机理提供可操作靶分子。 本论文采用高通量蛋白质组研究技术对小麦杂交坏死进行了研究。携带坏死基因Ne2的小麦品种Pan555（P）和携带坏死基因Ne2的小麦品种Zheng891（Z）生长发育完全正常，将两个亲本杂交，所得杂交F1代PZF1表现杂交坏死。在小麦生长阶段8，旗叶（Flag leaf）刚刚出现，PZF1的旗叶下第一片叶子（FL-1）还是完全绿色，FL-2叶尖开始有坏死斑出现。在这个阶段，分别将PZF1，P，Z的FL-2叶剪成相等的尖，中，基三段。我们选择的PZF1的FL-2叶，其叶尖段已经有成片的坏死斑出现;中间段零星出现少量坏死斑点;基部段和亲本一样还是完全的绿色，代表坏死进程中的不同阶段。又选PZF1的FL-1和FL-2分别代表杂交坏死启动前和杂交坏死启动后。两个亲本P和Z的FL-2叶的三段及FL-1叶正常，都是完全绿色。 首先分别分析了PZF1，P和Z的FL-2叶的尖、中、基三段的蛋白表达情况。在PZF1的尖、中、基三段共检测到23个差异表达蛋白点。这23个点在两个亲本的尖、中、基三段中的表达丰度没有显著差异（p<0.05），说明这23个蛋白的差异表达不是由于叶段的不同引起，确与杂交坏死相关。对这23个蛋白进行了MALDI-TOF质谱鉴定，其中18个得到成功鉴定。然后对PZF1，P和Z的FL-1叶和FL-2叶的蛋白表达情况进行了分析。与PZF1的FL-1叶比较，在FL-2叶中检测到19个蛋白上调，20个蛋白下调。这39个蛋白的丰度在两个亲本的FL-1和FL-2叶之间没有显著差异，说明这39个蛋白的差异表达不是由于叶位的不同引起，确与杂交坏死相关。对这39个蛋白进行质谱鉴定其中26个得到成功鉴定。 根据被鉴定蛋白的功能及其表达丰度的变化，对这些蛋白在小麦杂交坏死中可能的作用进行了讨论。与PZF1的FL-2叶基部相比，S-腺苷同型半胱氨酸水解酶（S-adenosyl homocysteine hydrolase）在中部极显著（p<0.01）下调，而在中部和尖段之间没有显著差异，保持低丰度不变。腺苷甲硫氨酸3（AdoMet synthase 3）和甲硫氨酸合成酶1（Methionine synthase 1）都在PZF1的FL-2叶尖段上调。甲基化循环中的这3个酶比例的不协调可能会以不同的方式加速细胞老化。 与PZF1的FL-1叶比较，尿卟啉环脱羧酶（Uroporphyrinogen decarboxylase）在FL-2叶中下调，这将引起尿卟啉环III的积累。脂加氧酶（Lipoxygenases）在FL-2叶中上调。尿卟啉环III的积累和脂加氧酶的上调都会引起细胞内活性氧的增加。另外活性氧和脂加氧酶都会使脂发生过氧化作用，进而导致细胞膜完整性受到破坏，最终可能导致细胞死亡。 与基部段比较，在PZF1的FL-2叶的尖段和/或中间段;以及与PZF1的FL-1叶比较，在FL-2叶中，都有很多防御性蛋白的上调，这暗示应对活性氧、脂过氧化、甲基化循环中三个酶比例的不协调等引起的对细胞的破坏作用，细胞可能启动了抗细胞死亡系统来应对这种细胞内部的胁迫。 然而，与基部段比较，一些能量相关蛋白在PZF1的FL-2叶的尖段和/或中间段;以及与PZF1的FL-1叶比较，在FL-2叶中的异常表达可能会以干扰能量循环的方式加速细胞死亡。另外，与FL-2基部段比较，在尖段和/或中间段，以及与PZF1的FL-1比较，在FL-2中，都有一些防御性蛋白、蛋白合成相关的蛋白以及单链DNA结合蛋白下调，它们的变化可能会降低细胞的抵抗力，蛋白合成能力以及DNA修复能力。细胞正常代谢的很多方面都受到干扰从而使PZF1叶细胞最终走向死亡。 本研究中发现了三个甲基化循环中的酶变化，而且S-腺苷同型半胱氨酸水解酶是在坏死进程的较早阶段发生下调，它的变化可能是小麦杂交坏死的一个诱因，这暗示小麦杂交坏死可能是一个表观遗传学事件。另外本研究还发现一些和活性氧，脂氧化等相关的蛋白的变化，而活性氧增加和脂氧化都是细胞凋亡的典型特征。所以本研究为表观遗传细胞凋亡和氧化胁迫细胞凋亡的研究提供了很有价值的信息。

New chiral E- and Z-isomers of the 1R,4R-2-arylidene-p-menthane-3-ones in induced cholesteric and ferroelectric liquid crystals

A novel pair of the E- and Z-isomeric 1R,4R-2-(4-heptyloxyphenyl)-benzylidene-p-menthan-3-ones has been prepared and the influence of distinctions in their molecular geometry on macroscopic properties of liquid crystal systems with the induced supra-molecular helical structure has been studied. The significantly lower helical twisting power of the chiral Z-isomer in comparison with that of E- one has been confirmed in the case of induced cholesteric systems based on 4-pentyl-4-cyanobiphenyl. The phase behavior and ferroelectric characteristics have been investigated for smectic-C* compositions based on the eutectic mixture of the homological 4-hexyloxyphenyl-4'-hexyloxy- and 4-hexyloxyphenyl-4'-octyloxybenzoates containing the novel isomeric chiral dopants. The spontaneous polarisation of the opposite signs induced by the isomeric chiral components has been revealed for the compositions studied. Distinctions in phase states, absolute values of the spontaneous polarization, smectic tilt angle and rotation viscosity of the systems obtained are discussed.

Ferroelectric liquid crystal mixtures containing chiral ether and ester compounds with the 2-arylidene-p-menthan-3-one skeleton

Some 1R,4R-2-(4-phenylbenzylidene)-p-menthane-3-one derivatives containing the ether or ester linking group between benzene rings of the arylidene fragment have been studied as chiral dopants in ferroelectric liquid crystal systems based on the eutectic mixture (1:1) of two phenylbenzoate derivatives CmH2m+1OC6H4COOC6 H4OCnH2n+1 (n = 6; m = 8, 10). The ferroelectric properties of these compositions (spontaneous polarization, rotation viscosity, smectic tilt angle as well as quantitative characteristics of their concentration dependences) were compared with those for systems including chiral dopants containing no linking group. Ferroelectric parameters of the induced ferroelectric compositions studied have been shown to depend essentially on the presence of the linking group between benzene rings and its nature as well as on the number of the benzene rings in the rigid molecular core of the chiral dopants used. For all ferroelectric liquid crystal systems studied, the influence of the chiral dopants on the thermal stability of N*, SmA and SmC* mesophases has been quantified. The influence of the linking group nature in the dopant molecules on the characteristics of the systems studied is discussed taking into account results of the conformational analysis carried out by the semi-empirical AM1 and PM3 methods.

Growth of Skipjack Katsuwonus pelamis from Mozambique

Growth of skipjack, Katsuwonus pelamis in Mozambican waters was studied, using fork length data collected in 1983-1985, during an experimental pole-and-line/live-bait fishing program. Tentative estimates of growth parameters are FL���=80 cm, K=0.6 yearˉ¹. Some evidence for seasonal growth is presented, along with a discussion of the limitations of the data set used in the investigation.

Economics of operation of 38' G.R.P. fishing vessels issued by the A.D.B. Fisheries Project: a case study

The author gives a financial and economic valuation of the operation of vessels in the 38 foot GRP class. He discusses particularly the materials and methods of fishing they use, species composition of their catches, their rate of return, break-even analysis, financial and social analysis.

New (1R,4R)-2-arylidene-p-menthan-3-ones with a bridging ester group in the arylidene fragment. Synthesis and behavior in liquid-crystalline systems

(1R,4R)-2-(4-Hydroxybenzylidene)- and (1R,4R)-2-(4′-hydroxybiphenyl- 4-yl)methylene-p-menthan-3-ones were synthesized by condensation of (-)-menthone with O-tetrahydropyran-2-yl derivatives of 4-hydroxybenzaldehyde and 4′-hydroxy-4-formylbiphenyl, respectively, in a DMSO - base medium followed by the removal of the protective group. The reactions of these hydroxy derivatives with 4-alkylbenzoic, 4-alkyloxybenzoic, trans-4-alkylcyclohexane-4- carboxylic, and 4′-alkylbiphenyl-4-carboxylic acids afforded three series of new chiral esters. Compounds containing the arylidene moiety with three benzene rings were found to exhibit liquid-crystalline properties. The characteristic features of these compounds are discussed based on the results of studies by polarizing microscopy, differential scanning calorimetry, and small-angle X-ray scattering. It was found that the mesomorphic compounds under study can form a smectic A mesophase, twist grain boundary mesophases (TGBA), and blue phases in a wide temperature range. Upon dissolution of certain of chiral compounds in 4′-cyano-4-pentylbiphenyl, a rather high twisting power and the thermal stabilizing effect on mesophases were observed.

New chiral ether derivatives of 2-arylidene-p-menthane-3-ones as components of induced ferroelectric systems

New 2-arylidene-p-menthane-3-ones containing the ether bridging group in the arylidene fragment have been synthesized and studied as chiral dopants in ferroelectric liquid crystal mixtures. The ferroelectric properties of these compositions were compared with those for compositions including chiral dopants that do not contain any bridging group. The influence of bridging group and terminal alkyl substituent length in the dopant molecule on the ferroelectric parameters of systems studied is discussed. © 2001 OPA (Overseas Publishers Association) N.V. Published by license under the Gordon and Breach Science Publishers imprint, a member of the Taylor & Francis Group,.

Effect of copper, cadmium and Cu.Cd mixture on amino acid content in the poslarvae of penaeid shrimp, Penaeus monodon and P. penicillatus

This study document effects of short-term (96h) sublethal levels of copper, cadmium and their mixture on the amino acid composition of postlarvae of the penaeid shrimp, P.monodon and P.penicillatus . All experimental conditions were kept constant, temperature between 25-27•C and salinity 21-22 ppt. The estimated LD50 for Cu was 200 ug/L, for Cd 177.5 ug/L and for Cu.Cd mixture 250ug/L. In P. penicillatus at the same concentration of each metal, there was significant reduction in amino acid content, which was 8.01% higher than the control. Almost similar reduction in some amino acids was observed in P.monodon. At the maximum concentration of 400 ug/L, cadmium caused higher reduction in amino acid composition than did copper. Thus, amino acid composition may be regarded as a sensitive biochemical indicator of Cu and Cd toxicity because of the effect of these metals on protein synthesis, a signal of physiological stress in marine organisms subjected to heavy metal pollution.

Cloning of two novel P-III class metalloproteinases from Trimeresurus stejnegeri venom gland

Hemorrhagic toxins are widely distributed in viperid and crotalid snake venoms. Envenomation of Trimeresurus stejnegeri, a member of Crotalidae family, caused potent systemic and local hemorrhage. Up to now, there is no report on hemorrhage toxins from th

A new protein structure of P-II class snake venom metalloproteinases: it comprises metalloproteinase and disintegrin domains

A new metalloproteinase-disintegrin, named Jerdonitin, was purified from Trimeresurus jerdonii venom with a molecular weight of 36 kDa on SDS-PAGE. It dose-dependently inhibited ADP-induced human platelet aggregation with IC50 of 120 nM. cDNA cloning and sequencing revealed that Jerdonitin belonged to the class II of snake venom metalloproteinases (SVMPs) (P-II class). Different from other P-II class SVMPs, metalloproteinase and disintegrin domains of its natural protein were not separated, confirmed by internal peptide sequencing. Compared to other P-II class SVMPs, Jerdonitin has two additional cysteines (Cys219 and Cys238) located in the spacer domain and disintegrin domain, respectively. They probably form a disulfide bond and therefore the metalloproteinase and disintegrin domains cannot be separated by posttranslationally processing. In summary, comparison of the amino acid sequences of Jerdonitin with those of other P-II class SVMPs by sequence alignment and phylogenetic analysis, in conjunction with natural protein structure data, suggested that it was a new type of P-II class SVMPs. (C) 2003 Elsevier Inc. All rights reserved.

High mobility N-channel and P-channel nanocrystalline silicon thin-film transistors

We report high hole and electron mobilities in nanocrystalline silicon (nc-Si:H) top-gate staggered thin-film transistors (TFTs) fabricated by direct plasma-enhanced chemical vapor deposition (PECVD) at 260°C. The n-channel nc-Si:H TFT with n+ nc-Si:H ohmic contacts shows a field-effect electron mobility (μnFE) of 130 cm2/Vs, which increases to 150 cm2/Vs with Cr-silicide contacts, along with a field-effect hole mobility (μhFE) of 25 cm2/Vs. To the best of our knowledge, the hole and electron mobilities reported here are the highest achieved to date using direct PECVD. © 2005 IEEE.

Studies on long wing trawl for shrimps off Kakinada (A.P.) coast - (i) Results of comparative fishing experiments with the conventional two seam trawl

Comparative fishing operations with the conventional two seams net and a 29.26 m. long wing shrimp trawl of four seam type were undertaken. The result showed that the four seams net gave nearly twice prawn catch than that of the conventional type. It was also found that the four seams net can be developed into a combination trawl for the effective exploitation of both prawns and fish along the coasts off Kakinada.