用AFLP技术进行水稻光合作用光保护有关基因候选分子标记的研究初探

本项目用扩增片段长度多型性(AFLP)技术,研究了水稻光合作用和光保护机制有关的8个性状F2代极端样本,在32对引物的研究中,全部引物可以在亲本中检出多型性,有28对引物可以在F2群体,8组极端样本之间找到差异条带标记,每个性状可以找到7～22个差异条带.在叶绿素ab含量、玉米黄素+花黄素总量、超氧化物歧化酶活性上,Lemont具有明显的增效片段优势.玉米黄素+花黄素+紫黄质数总量和活性蛋白总量可能是提高光合作用的最大限制因子.同时研究表明,对预选PCR 产物进行纯化处理,可大大提高AFLP分析技术的有效性.     

Bioactivity and characterization of exudates from Cryptomeria japonica bark

Tree exudates play an important role in the defense system of trees. In this study, the composition of exudates secreted from the wound site of Cryptomeria japonica was examined. Seven diterpenoids, including ferruginol (1), cryptojaponol (2), isopimaric acid (3), 7β-hydroxydeoxocryptojaponol (4), isopimarol (5), sandaracopimarinol (6) and phyllocladan-16α-ol (7), were identified through chromatographic separation and spectroscopic analyses. Among them, ferruginol was the most abundant compound, accounting for 76.6 % of the total content in exudates. Exudates, ferruginol and isopimaric acid possessed the strongest antibacterial activity and had MIC values of 25–250, 12.5–100 and 6.25–100 μg/mL, respectively, against one strain of Gram-negative bacteria (Escherichia coli) and four strains of Gram-positive bacteria (Staphylococcus aureus, methicillin-resistant S. aureus, S. epidermidis and Enterococcus faecalis). In addition, exudates and ferruginol presented 79 and 53 % at 100 μg/mL, respectively, of antioxidant activities against 1,1-dipheny1-2-picrylhydrazyl radical. C. japonica bark exudates and their diterpenes exhibit the strongest antioxidant activity and can inhibit bacterial growth and thus have the potential to be developed into natural antioxidants and bactericides.     

Effects of dietary supplementation of lipid‐encapsulated zinc oxide on colibacillosis,growth and intestinal morphology in weaned piglets challenged with enterotoxigenic Escherichia coli

This study was aimed to evaluate the effect of dietary supplementation of lipid‐encapsulated (coated) zinc oxide ZnO on post‐weaning diarrhea (colibacillosis) in weaned piglets challenged with enterotoxigenic Escherichia coli (ETEC). Thirty‐two 35‐day‐old weaned piglets were orally challenged with 3 × 10¹⁰ colony forming units of ETEC K88 while eight piglets received no challenge (control). Each eight challenged piglets received a diet containing 100 ppm ZnO (low ZnO), 2500 ppm ZnO (high ZnO) or 100 ppm of lipid (10%)‐coated ZnO (coated ZnO) for 7 days; control pigs received the low ZnO diet. Daily gain, goblet cell density in the villi of the duodenum, jejunum and ileum, and villus height in the jejunum and ileum, which decreased due to the challenge, were equally greater in the coated ZnO and high ZnO groups versus low ZnO group. Fecal consistency score, serum interleukin‐8 concentration, subjective score of fecal E. coli shedding, and digesta pH in the stomach, jejunum and ileum, which increased due to the challenge, were equally low in the coated ZnO and high ZnO groups versus low ZnO. Results suggest that a low level of coated ZnO might well substitute for a pharmacological level of native ZnO in dietary supplementation to alleviate colibacillosis of weaned piglets.     

Growth of uniformly oriented silica MFI and BEA zeolite films on substrates

Applications of zeolite films benefit from alignment of the integrated channels, but methods for film growth have nearly always introduced orientational randomization in the direction normal to the substrate. We now report facile methods to grow silicalite-1 films and pure silica beta zeolite films on substrates with straight or sinusoidal channels positioned uniformly upright at a thickness of up to 8 micrometers. Precise gel compositions and processing temperatures are critical to promote secondary growth on pre-formed oriented crystal monolayers while suppressing self-crystallization in the bulk medium. Preliminary results highlight the potential of these uniformly oriented films in the nonlinear optical response and separation of xylene isomers.     

Solvated electrons in high-temperature melts and glasses of the room-temperature stable electride [Ca₂₄Al₂₈O₆₄]⁴⁺·4e⁻

Solvated electrons in alkali metal-ammonia solutions have attracted attention as a prototype electronic conductor and chemical reducing agent for over a century. However, solvated electrons have not been realized in a high-temperature melt or glass of an oxide system to date. We demonstrated the formation of persistent solvated electrons in both a high-temperature melt and its glass by using the thermally stable electride [Ca(24)Al(28)O(64)](4+)·4e(-) (C12A7:e(-)) and controlling the partial pressure of oxygen. The electrical and structural properties of the resulting melt and glass differ from those of the conventional C12A7:O(2-) oxide, exhibiting metallic and hopping conduction, respectively, and a glass transition temperature that is ~160 kelvin lower than that of C12A7:O(2-) glass. Solvated electrons reside in cage structures in C12A7:e(-) and form a diamagnetic paired state.     

C-O bond formation by polyketide synthases

Polyketide synthases (PKSs) assemble the polyketide carbon backbone by sequential decarboxylative condensation of acyl coenzyme A (CoA) precursors, and the C-C bond-forming step in this process is catalyzed by the beta-ketoacyl synthase (KS) domain or subunit. Genetic and biochemical characterization of the nonactin biosynthesis gene cluster from Streptomyces griseus revealed two KSs, NonJ and NonK, that are highly homologous to known KSs but catalyze sequential condensation of the acyl CoA substrates by forming C-O rather than C-C bonds. This chemistry can be used in PKS engineering to increase the scope and diversity of polyketide biosynthesis.     

EMF, an Arabidopsis Gene Required for Vegetative Shoot Development

In higher plants, the transition from the vegetative to the; reproductive state in the shoot meristem initiates flowering. To study this floral transition, Constitutively flowering mutant of Arabidopsis thaliana ecotype columbia, embryonic flower (emf), was characterized. No vegetative shoots were produced from emf embryos or calli; the shoot apical meristems (SAMs) in the emf embryos were altered compared to wild-type SAMs. The mutant SAMs enlarged precociously and produced inflorescence meritems upon germination. These results suggest that the dominant, wild-type allele EMF is required for the vegetative state of the SAM. In the absence of EMF function, the mutant embryo assumes the reproductive state.     

菠菜胆碱单氧化酶(CMO)基因的克隆及在大肠杆菌中的诱导表达

甘氨酸甜菜碱是一种非毒性的渗透调节物质 ,在植物体内是以胆碱为底物 ,经两步氧化而合成的 .菠菜中 ,催化第一步反应的酶为胆碱单氧化酶 (CholineMonooxygenase ,CMO) .为了研究胆碱单氧化酶基因的功能以及转基因植物的抗逆能力 ,在 30 0mmol L高盐浓度 (n(NaCl)∶n(CaCl2 ) =5 7∶1)的条件下 ,作者分离纯化了菠菜mRNA ,经RT PCR得到全长 (1.3kb)胆碱单氧化酶 (CMO)cDNA ,与已经报道的基因序列相比较 ,同源性为 99% .根据其核苷酸序列推导得到了氨基酸序列 .将PCR纯化产物与pET 30a+ 连接 ,构建了重组表达载体pETCMO ,并转化到大肠杆菌BL2 1(DE3) ,经IPTG诱导获得高效表达 .     

Expression and characterization of sweet potato invertase in Pichia pastoris

An invertase cDNA (Ibbetafruct1) was cloned from sweet potato leaves and characterized. The deduced amino acid sequence of the Ibbetafruct1-encoded protein was closely related to vacuolar invertases and included the WECVD catalytic domain characteristic of them. An expression plasmid containing the coding region of Ibbetafruct1 under the control of the alcohol oxidase promoter was used to transform the methylotrophic yeast Pichia pastoris. The biochemical properties for the expressed recombinant enzyme, which was determined to be the acid beta-fructofuranosidase with an acidic pI value (5.1), were similar to those of vacuolar invertases purified from sweet potato. Periodic acid/Schiff staining and Con A-Sepharose gel-binding experiments revealed the recombinant invertase to be a glycoprotein containing glucose and/or mannose residues. Furthermore, the carbohydrate moiety appears to be a key determinant of the enzyme's sucrose hydrolysis activity, substrate affinity, and thermal stability.