陆地棉PIP亚家族基因的克隆及表达特征分析  

Cloning and Expression Analysis of PIP Subfamily Genes in Gossypium hirsutum

作　　者：孙雨薇[1] 李艳军[1] 刘永昌[1] 薛飞[1] 朱守鸿 王翔飞[2] 孙杰[1] 

机构地区：[1]石河子大学农学院新疆生产建设兵团绿洲生态农业重点实验室,新疆石河子832003 [2]石河子大学药学院,新疆石河子832003

出　　处：《西北植物学报》2016年第10期1948-1954,共7页Acta Botanica Boreali-Occidentalia Sinica

基　　金：国家自然科学基金(31460360)

摘　　要：该研究以陆地棉苯基香豆满苄基醚还原酶(phenylcoumaran benzylic ether reductase,PCBER)氨基酸序列为探针,利用Blastp从陆地棉基因组数据库中发现了6个同源性较高的基因。根据6个基因序列设计引物,利用RT-PCR技术从陆地棉纤维细胞中克隆出了这6个基因的全长cDNA序列,分别命名为GhPCBER1、GhPCBER2、GhPCBER3、GhIFR、GhPLR1和GhPLR2。多重序列比对和进化树分析发现,6个蛋白均含有PIP类型蛋白的所有保守性基序和活性残基,属于PIP亚家族。实时荧光定量PCR结果显示,除GhPLR1之外其他5个PIP亚家族基因均在纤维细胞中优势或特异表达;在纤维发育过程中,GhPCBER1、GhPCBER2、GhPCBER3和GhIFR的表达均表现为先上升后下降,GhPCBER1和GhPCBER2在花后21d表达量达到最高,GhPCBER3和GhIFR在花后18d达到最高,GhPLR1和GhPLR2在纤维中的表达量呈持续上升趋势。根据基因的表达特征,推测PIP亚家族可能在棉纤维的发育过程中发挥着重要作用。Using the protein sequence of GhPCBER as a probe, six of genes showing high sequence homolo gy were obtained from a genome database of Gossypium hirsutum L. tetraploid cotton with Blastp search. These sequences were used to design primers, and then six genes were isolated from cotton fiber of G. hir- suture L. by using RT-PCR technique and designated as GhPCBER1, GhPCBER2, GhPCBER3, GhIFR, GhPLR1 and GhPLR2, respectively. Multiple sequence alignment and phylogenetic tree analysis revealed that all of the 6 proteins belong to PIP subfamily since the conserved motifs and active residues in their se- quences. Real time fluorescent quantitative PCR results showed that in addition to GhPLR1, all the other 5 PIP subfamily genes were specifically or preferentially expressed in fiber cells. Of 6 genes, the expres- sion pattern of GhPCBER1, GhPCBER2, GhPCBER3 and GhlFR showed increasing firstly and then de- creasing trend during cotton fiber developing stage. The expression level of GhPCBER1 and GhPCBER2reached a maximum at 21 day post anthesis, and GhIFR and GhPCBER3 reached a maximum at 18 day post anthesis. The expression pattern of GhPLR1 and GhPLR2 showed an ascendant trend with cotton fi- ber developing. According to the characteristics of gene expression, it is speculated that the PIP subfamily may play an important role in cotton fiber development.

关 键 词：棉花 纤维 PIP 表达分析 

分 类 号：Q785[生物学—分子生物学] Q789