蛇苔过氧化物酶基因的序列分析与功能预测

过氧化物酶在生物界分布极广,在细胞代谢的氧化还原过程中起重要的作用.为深入研究蛇苔过氧化物酶的理化性质及功能,对其同源性,亚细胞定位,结构和功能等进行了生物信息学预测和分析.结果表明;该序列有1 050 bp长的开放阅读框,编码349个氨基酸,与拟南芥等的POD相似性较高(相似性为71％).预测R68、F71、H72、...     

蛇苔细胞色素C（Cyt C）的序列分析及功能预测

通过核酸序列比对,在蛇苔cDNA文库中获得细胞色素C（Cyt C）基因序列,并对其编码的蛋白质产物从同源性、氨基酸组成、理化性质、亚细胞位点、结构和功能等进行生物信息学分析和预测。结果表明,该cDNA序列具有完整的开放阅读框（ORF,104—442bp）,推测编码蛋白为112个氨基酸,与向日葵、荞麦、玉米等Cyt C存在着较高的保守性（相似性分别为84％、85％和84％）,同属于细胞色素C超家族;蛇苔CytC蛋白的分子量为11998．7Da,不含信号肽,成熟的CytC起始于Ala2,其活性位点为Cys23、Cys26、His27和Met89;对蛋白质的翻译后修饰预测表明Ala2存在乙酰化修饰,Lys81和Lys95分别进行三甲基赖氨酸修饰。这些特点与其他植物CytC保持一致,表明该基因为蛇苔cyt c基因,在进化上相对保守。     

广义柏科主要分类群起源时间的推测

通过构建分子钟对广义柏科主要分类群的起源时间进行探讨。采用相对速率检验法分析广义柏科mat K、rbc L进化速率的稳定性,结果显示rbc L的非同义替代速率只在Pinaceae与Taxodiaceae的分类群及柏科北半球的支系(Cupressoideae)之间通过相对速率检验,而Pinaceae与柏科南半球分支(Callitrodeae)之间没有通过相对速率检验。mar K基因的非同义替代速率在Pinaceae与广义柏科的所有分类群之间通过相对速率检验。根据通过相对速率检验的分类群之间的遗传距离和基因进化速率,计算它们发生分歧的时间。据此推测,杉科的主要分类群Taiwanioideae、Athrotaxidoideae、Sequoioideae与其他分支发生分歧的时间均在侏罗纪,支持现存杉科在侏罗纪就已经建立起来的观点;Cupressaceae(s．s．)的两个支系(亚科)发生分歧的时间在124Ma之前,相当于早白垩世早期,可能由于南北古大陆的完全分离,其祖先居群被分隔成两个亚群,随后各自演化为不同的支系。Callitirodeae、Cupressoideae各属发生分歧的时间也均在白垩纪,表明Cupressaceae(s．s．)在白垩纪就已经建立起来。     

不同理化因子对雪莲毛状根生长和总黄酮生物合成的影响

在 1 2MS液体培养基上研究了不同理化因子对水母雪莲毛状根生长和总黄酮生物合成的影响。实验结果表明 :氮源总浓度 (包括NH+4和NO-3)为 30mmol L ;NH⁺₄ NO^-₃比例为 5∶2 5 ;2 %蔗糖和 3%葡萄糖组合 ;0.5mg LGA₃和 0.5mg LIBA ;pH5.8;18h d的光照 (光强为 35.0.0lx) ;2.4℃ ;摇床转速为 100rmin有利于毛状根生长及总黄酮的生物合成。在此培养条件下 ,经过21d的培养毛状根生长量达到 12.8g L(DW) ,总黄酮合成量为 192.2mg L ,即总黄酮含量占毛状根干重的 15 % ,约为干重野生水母雪莲植株总黄酮含量的 2.5倍。     

水母雪莲愈伤组织cDNA文库的构建

用TRIZOL Reagent提取水母雪莲红色系1~15 d愈伤组织总RNA,用SMART cDNA Library Construction Kit构建cDNA文库。经测定原始文库滴度达到1.5~4×106,扩增总文库滴度达到1011,重组率达到98%,插入片段在0.5 kb到3 kb之间,多在1 kb左右。通过PCR检测,从总文库中检测到了雪莲CHS、DFR及SmP基因的特异片段。SmP基因是转录调控因子,其表达丰度很低。各项指标都表明,已获得高质量的cDNA文库,为雪莲基因资源保存,雪莲类黄酮次生代谢分子调控奠定了坚实的基础。     

杉科、柏科的系统发生分析——来自28S rDNA序列分析的证据

对杉科（Taxodiaceae）与柏科（Cupressaceae s.s.）的28S rRNA基因的部分序列（约630 bp）进行PCR扩增、序列测定和系统发生关系分析,用简约法和邻接法构建的系统发生树基本一致。结果表明,杉科与柏科构成一个单系群,支持将杉科、柏科（Sciadopitys除外）合并为一个科——广义柏科（Cupressaceae sensu lato）的观点。在广义柏科中,Taiwania、Athrotaxis分别形成一支系;Metasequoia、Sequoia、Sequoiadendron关系较近,聚成一支系; Taxodium、Glyptostrobus、Cryptomeria聚成一支系;柏科聚成一支系。这一分析结果与叶绿体基因序列的分析结果相吻合,但是由于28S rRNA基因的进化速率较慢,尚不能分辨上述各个支系之间的系统演化关系。 Abstract:DNA sequences from 28S rDNA were used to assess relationships between and within traditional Taxodiaceae and Cupressaceae s.s.The MP tree and NJ tree generally are similar to one another.The results show that Taxodiaceae and Cupressaceae s.s.form a monophyletic conifer lineage excluding Sciadopitys.In the Taxodiaceae-Cupressaceae s.s.monophyletic group,the Taxodiaceae is paraphyletic.Taxodium,Glyptostrobus and Cryptomeria forming a clade(Taxodioideae),in which Glyptostrobus and Taxodium are closely related and sister to Cryptomeria;Sequoia,Sequoiadendron and Metasequoia are closely related to each other,forming another clade (Sequoioideae),in which Sequoia and Sequoiadendron are closely related and sister to Metasequoia;the seven genera of Cupressaceae s.s.are found to be closely related to form a monophyletic lineage (Cupressoideae).These results are basically similar to analyses from chloroplast gene data.But the relationships among Taiwania,Sequoioideae,Taxodioideae,and Cupressoideae remain unclear because of the slow evolution rate of 28S rDNA,which might best be answered by sequencing more rapidly evolving nuclear genes.     

The low density lipoprotein receptor-related protein (LRP) is a novel beta-secretase (BACE1) substrate

BACE is a transmembrane protease with beta-secretase activity that cleaves the amyloid precursor protein (APP). After BACE cleavage, APP becomes a substrate for gamma-secretase, leading to release of amyloid-beta peptide (Abeta), which accumulates in senile plaques in Alzheimer disease. APP and BACE are co-internalized from the cell surface to early endosomes. APP is also known to interact at the cell surface and be internalized by the low density lipoprotein receptor-related protein (LRP), a multifunctional endocytic and signaling receptor. Using a new fluorescence resonance energy transfer (FRET)-based assay of protein proximity, fluorescence lifetime imaging (FLIM), and co-immunoprecipitation we demonstrate that the light chain of LRP interacts with BACE on the cell surface in association with lipid rafts. Surprisingly, the BACE-LRP interaction leads to an increase in LRP C-terminal fragment, release of secreted LRP in the media and subsequent release of the LRP intracellular domain from the membrane. Taken together, these data suggest that there is a close interaction between BACE and LRP on the cell surface, and that LRP is a novel BACE substrate.     

Nitrate-inducible MdBT2 acts as a restriction factor to limit apple necrotic mosaic virus genome replication in Malus domestica

Apple necrotic mosaic virus (ApNMV) is highly associated with the occurrence of apple mosaic disease in China. However, ApNMV–host interactions and defence mechanisms of host plants against this virus are poorly studied. Here, we report that nitrate treatment restrains ApNMV genomic RNA accumulation by destabilizing viral replication protein 1a through the MdBT2-mediated ubiquitin-proteasome pathway. MdBT2, a nitrate-responsive BTB/TAZ domain-containing protein, was identified in a yeast two-hybrid screen of an apple cDNA library using viral protein 1a as bait, and 1a was further confirmed to interact with MdBT2 both in vivo and in vitro. It was further verified that MdBT2 promoted the ubiquitination and degradation of viral protein 1a through the ubiquitin-proteasome pathway in an MdCUL3A-independent manner. Viral genomic RNA accumulation was reduced in MdBT2-overexpressing transgenic apple leaves but enhanced in MdBT2-antisense leaves compared to the wild type. Moreover, MdBT2 was found to interfere with the interaction between viral replication proteins 1a and 2a^pol by competitively interacting with 1a. Taken together, our results demonstrate that nitrate-inducible MdBT2 functions as a limiting factor in ApNMV viral RNA accumulation by promoting the ubiquitination and degradation of viral protein 1a and interfering with interactions between viral replication proteins.     

Adaptive plasticity in response to light and nutrient availability in the clonal plant Duchesnea indica

克隆植物蛇莓对光照强度和养分条件的适应性可塑性 表型可塑性可帮助植物缓冲环境压力并使其表型与当地环境相匹配,但目前仅少数性状的可塑性被广泛认为是适应性的。为充分理解可塑性的适应性意义,仍需进一步研究更多的植物功能性状及其环境因子。本研究将匍匐茎克隆植物蛇莓(Duchesnea indica)的21个基因型种植于不同的光照和养分条件下,并利用选择梯度分析检测了形态和生理可塑性对光照强度和养分有效性变化的适应性值。在遮荫条件下,蛇莓适合度(果实数、分株数和生物量)降低,节间缩短变细,成熟叶叶绿素含量降低,但叶柄长度、比叶面积、老叶叶绿素含量均增加。在低养分条件下,植株叶柄缩短,叶面积缩小变厚,叶绿素含量降低,但果实数量和根冠比增加。选择梯度分析表明,叶柄长度和老叶叶绿素含量对光照变化的可塑性是适应性的,老叶和成熟叶叶绿素含量对养分变化的可塑性也是适应性的。因此,不同性状的可塑性适应值取决于特定的生态背景。该研究的发现有助于理解克隆植物表型可塑性响应环境变化的适应性意义。