基于白掌株型突变转录组差异的SSR位点分析

Analysis of SSR Loci Based on Plant-type Mutant Transcriptome Differences in Spathiphyllum kochii

为了解白掌株型突变的转录组序列功能和SSR位点分布特征,并探索与株型关联的SSR位点信息。以对照‘美酒’及其小株型突变体为材料,选取叶片、叶柄、佛焰苞片、苞梗组织进行高通量PE150的转录组测序和序列筛选;开展unigene的GO、KOG、KEGG功能注释及分类;开展SSR位点筛选及批量设计引物;发掘差异表达含SSR位点unigene的GO、KEGG富集功能信息。结果显示,白掌转录组共筛选127153个unigene,其中有41370个获注释,包含GO注释25925个、KO注释12727个、KOG注释21487个;从18636个unigene中发掘出28046个SSR位点,出现频率22.06%,包含有193种重复基序,其中以二核苷酸AG/CT(14305,51.01%)、AC/GT(3907,13.93%)、AT/AT(2322,8.28%)和三核苷酸AAG/CTT(2087,7.44%)居多,基序重复次数以5~10次重复占优势;从18636个unigene中筛选17070个批量设计出51210对SSR引物,有效扩增和多态性好。差异转录组结合SSR位点信息显示,在与株型相关的4个组织中共获得1578个差异表达的unigene,其中有552个具SSR位点,占总SSR 2.96%、差异基因中的SSR 34.98%。KEGG富集分析筛选了26个含SSR位点的显著性差异unigene参与植物激素信号传导,脂肪酸延长,角质、琥珀和蜡的生物合成,甘露糖型O-聚糖的生物合成,苯丙素的生物合成,二萜生物合成等6个代谢途径。本研究为白掌株型变化的分子机制研究提供理论依据。

In order to understand the transcriptome sequence function and SSR locus distribution characteristics of Spathiphyllum kochi plant type mutation,and to explore the information of SSR locus associated with plant type.The control‘Meijiu’and its plant-type mutants were used as materials,tissue samples of leaves,petioles,spathes,and bracts were selected to transcriptome sequencing by Illumina PE150.Moreover,the unigene squencess were screened.The unigenes of transcriptome were annotated by aligning with several public databases including GO,KOG,and KEGG used BLAST program.The SSRs loci were examined,and the PCR amplification SSR primers were designed.The enrichment function information of GO and KEGG differentially expressed unigene containing SSR loci were explored.The results showed that a total of 127153 unigenes were screened from Spathiphyllum kochi transcriptome,of that 41370 were annotated,including 25925 GO annotations,12727 KO annotations and 21487 KOG annotations.Furthermore,28046 SSR loci were identified from 18636 unigenes that had 193 repeat motifs.The frequency of SSR was 22.06%.Among all the SSR motifs,the dinucleotide AG/CT(14305,51.01%),AG/CT(3907,13.93%),AC/GT(2322,8.28%)and trinucleotide AAG/CTT(2087,7.44%)were the most frequent motifs,and the repeat numbers from 5 to 10 of SSR were the most repeat numbers of motifs.Moreover,17070 unigenes were screened from 18636 unigenes,and 51210 pairs of SSR primers were designed.The SSR primer amplification and polymorphism were better.The transcriptome combined with SSR loci results showed that 1578 differentially expressed unigenes were obtained in four tissues related to the plant type of Spathiphyllum kochi,among them 552 contained SSR loci.The proportion of differential SSRs in the total SSRs was 2.96%,and the proportion of SSRs in the differential genes was 34.98%.KEGG enrichment analysis showed that there were 26 different unigenes containing SSR loci involved in six metabolic pathways,including phytohormone signaling,fatty acid elongation,biosynthesis of cutin,amber and wax,biosynthesis of mannose type O-glycan,biosynthesis of phenylpropanoid,and biosynthesis of diterpene.This study provides a theoretical basis for the study of molecular mechanism of the type change of Spathiphyllum kochi.