蜈蚣草磷酸盐转运蛋白的生物信息学分析与表达模式鉴定及PvPT3功能初步研究
毕业论文关键词：蜈蚣草 磷酸盐转运蛋白 As V PvPht1;3
Bioinformatics analysis and expression pattern identification of phosphate transporters in Peteris vittata and functional preliminary identification of PvPT3
Abstract：Peteris vittata is an arsenic hyperaccumulator which arsenic concentration in fronds is 2 percent of the dry fern。 Arsenate (As V) is the main arsenic species in soils and the main fern-available form of arsenic in the nature environment。 Arsenate was absorbed via the phosphate (Pi) transporters in soils for the similar chemical structure between Pi and As V。 Here, PvPht1; 1-6 are identified from the transcriptome database of Peteris vittata。 Employing qRT-PCR and bioinformatics analysis, expression pattern and bioinformatics characteristics of PvPht1; 1-6 have been determined。 The results showed that PvPht1;3 and PvPht1;6 transcripts were induced by AsV, which results in a significant up-regulation of the expression of PvPht1;3 by 80-fold and PvPht1; 6 by 5-fold under 20μM arsenic treatment。 Using PvPht1; 3 -overexpressing tobaccos, we investigated the impacts of overexpressing PvPht1; 3 on As V uptake and translocation。。 In this study, overexpression of PvPht1; 3 increased the total arsenic concentration of the shoots 2- to 4-fold compared with wild-type plants。 Arsenic transfer coefficient from root to shoot was 2- to 5-fold higher than wild-type plants。 It suggested that In a word, PvPht1; 3 overexpression enhanced arsenic uptake and root to shoot translation in transgenic tobacco。 PvPht1; 3 was involved in the arsenic efficient uptake and root to shoot translation in Peteris vittata。
Key words: Peteris vittata; phosphate transporters; As V; PvPht1;3
目 录
摘要1
关键词1
Abstract1
Key words1
引言2
1材料与方法 3
1。1 生物信息学分析 3
1。1。1 蜈蚣草、水稻、拟南芥磷酸盐转运蛋白家族成员核苷酸序列的获得 3
1。1。2 PvPht1家族基因多序列比对、进化树构建及蛋白结构域分析3
1。2 表达模式分析3
1。2。1试验材料3
1。2。2植物生长条件及采样3
1。2。3 植物总RNA提取、cDNA合成及RT-PCR反应4
1。3 过量表达PvPht1；3对烟草砷积累的影响 4
1。3。1 试验材料 4
1。3。2 转基因苗的鉴定 4
1。3。3 植物生长条件及采样 4
1。3。4 植物总砷含量测定 4
2结果与分析 4
2。1蜈蚣草中高亲和磷转运蛋白基因PvPht1的生物信息学分析 4
2。1。1 序列一致性分析及进化树构建 4
2。1。2 氨基酸队列分析及蛋白功能结构域分析 6
2。2 PvPht1 家族成员表达水平对砷胁迫的响应7
2。3 PvPht1；3过量表达对转基因烟草材料砷吸收转运的影响8
2。3。1 PvPht1；3 过量表达转基因烟草材料的筛选及鉴定 8
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