Characterization of threerice CCoAOMT genes

Caffeoyl-Coenzyme A 3-O-Methyltransferase (CCoAOMT) is a key enzyme in lignin biosynthesis pathway. Three rice CCoAOMT genes were identified and designated as OsCOAI, OsCOA20 and OsCOA26. OsCOAI contains four exons and three introns, while the other two have three exons and two introns. The deduced amino acid sequences of these rice genes share a high identity (75.43%) with other plant CCoAOMT proteins and contain the CCoAOMT specific motifs. Phylogenetic analysis indicates that OsCOAI has the closest evolutionary relationship to maize CCoAOMT. In contrast, OsCOA20 and OsCOA26 belong to another clade.Northern blot analyses and in situ hybridization studies indicate that the three rice CCoAOMT genes are highly expressed in developing sclerenchyma cells and vessel bundles of young leaves, suggesting that they are probably involved in constitutive lignification.     

Obtaining the transgenic poplars with low lignin content through down-regulation of 4CL

The antisense 4CL (4-coumarate: CoA ligase)gene was transformed into triploid Chinese white poplar(Populus tomentosa) mediated by Agrobacterium tumefaciens.PCR and Southern blot analysis indicated that antisense 4CLgene had been integrated into the genome of the transgenicChinese white poplars. The antisense gene had also beenexpressed, which was indicated by RT-PCR and Westernanalysis. Klason lignin content assay showed that repressionof 4CL expression could result in remarkable reduction oflignin content in transgenic poplars, with most reduction of 41.73% compared with that of wild type in this paper. Butthere is no significant difference in holocellulose content be-tween trans- genic and wild poplars. We considered that 4CLmight not be the metabolism control point between ligninand carbohy- drate biosynthesis. The stems of transgenicpoplars displayed red-brown color with different levels afterthe bark was peeled, while those of untransformed plantswere white. No visible differences in growth and developmentwere observed between transgenic and wild plants. Wiesnerreaction analysis of the transgenic plant stems with reducedlignin content exhibited red color, while that of untrans-formed plant was typically purple-red.     

转基因抗虫棉花木质素含量及其生物合成关键酶活性研究

通过盆栽试验,以不同转基因抗虫棉花及其亲本对照为供试对象,研究木质素含量及其生物合成关键酶活性.结果表明:与非转基因棉花比较,在蕾期及铃期,转基因抗虫棉花Z30,CCRI41,SGK321叶片、茎秆组织中的木质素含量及叶片内的苯丙氨酸解氨酶和过氧化物酶活性发生了一些变化,但其显著性改变仅特异性地发生在某些品种、某些组织或某些时期内.在蕾期,转Bt基因棉花Z30茎秆组织中的木质素含量比其常规棉对照Z16显著下降,幅度达34.96%;转CpTI-Bt棉花CCRI41茎秆组织和SGK321叶片组织中的木质素含量比其常规棉对照显著下降,幅度分别为27.15%,13.22%.所有供试转基因抗虫棉花的木质...     

两性木质素絮凝剂的制备及脱色性能研究

木素磺酸钙与丙烯酰胺接枝共聚物通过Mannich反应,得到两性木素絮凝剂LSM.由正交实验得到Mannich反应较佳条件为:醛胺比1∶1,反应温度50℃,反应时间2.0 h,在此条件下LSM具有较佳的脱色性能.并将LSM用于活性艳蓝X-BR、活性黄X-R、活性紫K-3R、活性黑K-BR、直接橙S等多种模拟染料废水的脱色处理,讨论了LSM投加量和溶液pH值对不同染料脱色效果的影响.实验结果说明,LSM对各染料有较好的脱色效果,脱色率均在82%以上.同时投加量增加可能引起废水CODcr的提高,需控制适当的投加量范围.     

白腐菌对秸秆中木质素生物降解的研究

研究了白腐菌及其产生的木质素降解酶系对秸秆的木质素生物降解方法，探讨了黄孢原毛平革菌和杂色云芝单一菌株生物降解及双菌株联合降解木质纤维素的规律，白腐菌双菌联合固态培养可使木质素降解率达到47．64％，脱木素选择性为4．74．采用模拟大规模白腐菌堆积培养（厚度30cm）的方法降解秸秆木质素，培养30d，木素降解率为32．54％，表明此方法是一种可行的大规模生物顸处理方法．     

假单孢菌PKE117木素过氧化物酶的酶学性质及基因克隆

对纯化得到的假单孢菌PKE117的胞外木素过氧化物酶的酶学性质进行了初步研究,发现其最适pH为5.0,最适温度为30℃,以藜芦醇为底物,在25℃,pH3.0的琥珀酸钠缓冲液中与底物反应时的Km值为(0.277±0.0008)mmol/L,vmax为(0.049±0.001)mmol.mg-1protein.min-1。根据LiP已知保守序列设计引物,提取质粒,扩增得到一条199bp的DNA片段lip1和一条563bp的DNA片段lip2,序列比对结果显示与已报道的白腐真菌过氧化物酶基因的同源性不是很高。核苷酸翻译后的氨基酸序列比对结果发现,LiP1与Pycnoporus sanguineus的漆酶的同源性达到100%,与Mycobacteriumbovis的木素过氧化物酶的同源性达到80%;LiP2与Arabidopsis thaliana的过氧化物酶同源性达到100%,与Hordeum vulgare的过氧化物酶1同源性为75%。     

落叶松和水曲柳木质素热降解机理对比

用TGA,FTIR和XPS等手段对落叶松木质素和水曲柳木质素凝聚相的热降解过程及成炭行为进行对比研究。TGA分析表明:在高温纯氮条件下,落叶松木质素比水曲柳木质素剩余更多的炭残渣,这是由于落叶松木质素中的C-C键较多,使得落叶松木质素的碳骨架比水曲柳木质素的碳骨架更稳定。在空气气氛中,木质素在高温时完全分解为挥发性物质,这两种树的木质素均没有炭残余物,主要是由于在氧的催化作用下,C-C键和C-H键发生了氧化反应,这已经被FTIR实验证明。FTIR和XPS分析表明,在纯氮气气氛中,脂肪族酯键发生断裂,在凝聚态生成更多的苯环。根据C1s总强度和C1s(C-C)的相对强度以及碳/氧比例的显著提高得出,水曲柳木质素交联速度较大。     

金针菇降解木质素的能力及相关酶活研究

通过对10个金针菇菌株研究发现,在液体培养下不同菌株降解木质素的能力有较大差异,其中金针菇J89-6具有一定的降解木质素的能力,降解率达46%.本文还测定了不同菌株的漆酶、木质素过氧化物酶和锰过氧化物酶三种木质素降解酶的酶活,结果显示漆酶的产量与木质素降解率呈正相关关系.     

丙烯酰胺与木质素磺酸盐共聚接枝的研究

在常规实验条件下木质素磺酸盐与丙烯酰胺能发生接枝共聚。研究了聚合影响因素,得到木质素与水的质量比,木质素与单体的质量比,反应温度,引发剂浓度、反应时间等因素对改性反应影响的最佳配比条件。-CONH2接枝链的产生削弱了木质素原有的网状结构,因此接枝后的产物吸附能力较改性前明显增强。     

木质素的胺化改性

采用Ｍａｎｎｉｃｈ反应使木质素接枝胺基,生成木质胺阳离子表面活性剂,应用元素分析仪测定改性样品的含氮量。并通过三因子一次回归正交设计安排试验,得到影响含氮量的主要因素和最佳佳工艺条件。