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国家自然科学基金(31070576)

作品数:6 被引量:19H指数:2
相关作者:宋福强冯乐金涛周东坡刘盛春更多>>
相关机构:黑龙江大学东北林业大学教育部更多>>
发文基金:国家自然科学基金哈尔滨市科技创新人才研究专项资金项目中国博士后科学基金更多>>
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紫穗槐菌根蛋白双向电泳分离体系的构建被引量:1
2012年
丛枝菌根(arbuscular mycorrhizal,AM)真菌与宿主植物紫穗槐形成菌根过程中,双方都会发生一系列复杂的形态学和生理学变化,是一个多方面参与并精细调控的信号事件。通过温室盆栽试验,运用蛋白质组技术研究了AM真菌侵染紫穗槐过程中产生的共生相关蛋白,采用改良酚提取法提取紫穗槐菌根蛋白,并对蛋白上样量、染色方法等进行优化。结果表明,采用改良酚提取法,选用24 cm pH值4~7的胶条、1 000μg上样量、考马斯亮蓝R-350染色可获得分辨率高、重复性好的双向电泳图谱;应用凝胶分析软件Image Master TM 2D Platinum(Version 7.0)分析紫穗槐AM真菌侵染率达到90%时的菌根蛋白,接种AM真菌处理的GM组共获得434个蛋白点,而作为对照的CK组共获得419个蛋白点。其中GM组蛋白点与CK组相比较,上调表达蛋白点14个,下调表达蛋白点3个,特异表达蛋白点15个。
宋福强王健孔祥仕常伟王倡宪
关键词:紫穗槐丛枝菌根真菌双向电泳
Transcriptome analysis of the symbiosis-related genes between Funneliformis mosseae and Amorpha fruticosa
2019年
Arbuscular mycorrhizal fungi(AMF) can colonize and form associations with the roots of Amorpha fruticosa L.(desert false indigo). Various genes are induced during the symbiotic process. In this study, de novo transcriptome sequencing using RNA-seq was conducted for the first time for a comprehensive analysis of AMF-A. fruticosa symbionts at the transcript level. We obtained 12 G of raw data from illumina sequencing and recovered 115,786 unigenes with an average length of547 bp, among them 41,848 of significance. A total of2460 diffexpression genes were identified, including 1579 down-regulated and 881 up-regulated genes. A threshold for false discovery rate of \ 0.001 and fold change of [ 1 determined significant differences in gene expression.Using these criteria, we screened 285 significant differentially expressed genes, of which 82 were up-regulated and203 down-regulated. The 82 up-regulated genes were classified according to their functions and assigned into seven categories: stress and defense, metabolism, signaling transduction, protein folding and degradation, energy,protein synthesis, and transcription. The 203 down-regulated genes were screened according to fold change [ 2,and 50 highly significant down-regulated genes were obtained related to stress and defense. The results of this study will provide a useful foundation for further investigation on the metabolic characteristics and molecular mechanisms of AMF associations with leguminous woody shrubs.
Xuan LiuMingguo JiangFuqiang Song
关键词:ARBUSCULARMYCORRHIZALFUNGIRNA-SEQQUANTITATIVE
Characterization of expressed genes in the establishment of arbuscular mycorrhiza between Amorpha fruticosa and Glomus mosseae被引量:1
2014年
Arbuscular mycorrhiza (AM) formed between plant roots and fungi is one of the most widespread symbiotic associations in nature. To understand the molecular mechanisms of AM formation, we profiled 30 symbiosis-related genes expressed in Amorpha fruticosa roots colonized by Glomus mosseae and in non-mycorrhizal roots at different stages using differential-display RT-PCR (DDRT-PCR). The expressed genes were confirmed by reverse Northern blotting. Eleven fragments were sequenced and putatively identified by homologous alignment. Of the eleven AM-related genes, five were obtained at the early-stage of plant-fungus interaction and six at the later stage. Three expressed se-quence tag (ESTs) sequences were found to originate from the fungi and eight from the host plant by use of PCR evaluation of gDNA of both plant and fungi. The target genes included an ATP-binding cassette sub-family transporter gene, a transposon-insertion display band, and a photosynthesis-related gene. The results provided information on the molecular mechanisms underlying the development of mycorrhizal sym-biosis between woody plants and AM fungi.
Fuqiang SongJize LiXingxing Zhang
紫穗槐AfUGPase基因的生物信息学分析被引量:2
2015年
旨在了解UGPase基因的结构和功能,探明UGPase基因在AM真菌胁迫下,接种AMF诱导的紫穗槐植株与未接种AMF紫穗槐植株之间的差异。利用生物信息学方法如核酸序列比对分析、ORF分析、多序列比对和进化树分析等方法对该基因的核苷酸组成、蛋白质结构、系统进化发育等方面进行预测和分析。本研究得到Af UGPase基因全长为1831 bp,包含1个1416 bp的开放阅读框,编码471个氨基酸,蛋白分子量为51584.2 Da,等电点为6.07。是一种稳定蛋白,不含跨膜区,蛋白质的二级结构以α螺旋和无则卷曲为主,亚细胞定位预测显示该蛋白主要位于细胞质中,系统进化分析显示与大豆的UGPase在一个次级分化群。通过预测发现Af UGPase基因含有保守的Lys残基,参与酶的催化作用,是典型的UGPase蛋白,并在植物糖代谢、脂类调控合成中发挥重要作用。
刘璇宋福强齐丹丹高扬于美迪赵天琦孙玉芳
关键词:紫穗槐生物信息学
UPLC-ESI-MS/MS法测定发酵液中紫杉醇含量被引量:3
2012年
产紫杉醇菌发酵液经乙酸乙酯萃取后,应用Waters ACQUITY UPLC仪和Waters Quattro Premier三级四极杆质谱仪检测建立了产紫杉醇菌发酵液中紫杉醇含量的快速测定方法。采用Waters zXTerra MS C18色谱柱(2.1 mm×150 mm,5μm),梯度洗脱,流速0.3 mL/min,柱温为30℃;进样量3.0μL。以超高效液相色谱分离、电喷雾离子化串联质谱分析在多反应监测(MRM)模式下进行检测。在10~100μg/mL,紫杉醇对照品的峰面积与浓度线性关系良好,相关系数0.998;方法具有良好的精密度,RSD为0.51%;稳定性试验表明室温下样品在8 h内稳定;具有良好的重现性,RSD为1.6%;平均加样回收率为99.58%,RSD为0.62%。采用MRM模式测定紫杉醇含量不仅可以增加定量准确性,而且专一性强,干扰因素少,还为产紫杉醇微生物的快速筛选提供科学的方法。
金涛刘盛春周东坡平文祥宋福强阎秀峰
关键词:MS发酵紫杉醇
外生菌根真菌与丝状真菌混合对红松凋落物降解效能的影响被引量:12
2011年
实验以原始红松林中高频出现的丝状真菌(链格孢Alternarria sp.)和两种外生菌根真菌(细粉绒牛肝菌.XerocomusPulverulentus、大杯伞Clitocybe maxima)为供试菌株,以粉碎的新鲜红松凋落物为分解底物,发酵培养后测定底物的质量损失率,酶活性以及营养元素的变化情况。结果表明,AX处理(Alternaria sp.和X.PulveRrulentus混合菌株组合)在60天内质量损失率最高,为18.33%,而混合菌AC处理(Alternaria sp.和C.maxima混合菌株组合)由于产生拮抗作用质量损失较低,为13.27%;20天时,X处理(X.Pulverulentus)产生漆酶活性较高为0.093 U·mL^(-1),C(C.maxima)和AX处理的酶活性的最高值分别为0.063 U·mL^(-1)和0.047 U.mL^(-1),而A(Alternaria sp.)处理的纤维素酶活性为0.59 U·mL^(-1),AC处理为0.57 U·mL^(-1),AX为0.53 U.mL^(-1),但是AX处理在50天时再次出现高峰,为0.48 U·mL^(-1):AX处理底物的营养元素N含量减少幅度最为明显,减少百分率为12.21%,同时,P元素的减少幅度也最大,减少百分率25.82%。研究结果进一步揭示了森林生态系统中不同功能真菌类群对林木有机凋落物降解过程中的作用机制,及真菌类群间的相互作用关系。
冯乐宋福强
关键词:外生菌根菌丝状真菌凋落物降解率
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