利用转录组测序分析茄子萼片刺相关差异基因

Analysis of Prickle Development Genes in Eggplant Calyx via Transcriptome Sequencing

为了发掘茄子萼片刺形成的相关基因,为茄子萼片刺形成的机理和基因定位奠定基础,以萼片有刺和萼片无刺的2个自交系1607和1608为材料,构建了F2遗传分离群体,利用新一代高通量测序手段对双亲和2个混池的转录组进行了测序分析。通过转录组分析,共获得409363358个干净的高通量序列,其中比对到参考基因组的序列共有311756616个,比对效率均大于75.00%。发掘24310个新基因,其中获得注释信息的新基因总数为1540个。对转录组测序获得的全部基因表达量进行了分析,在双亲中共得到2438个差异表达基因,其中97个差异基因在2个混池中同时存在。通过对差异基因的注释分析,富集到了可能与茄子萼片刺形成的相关的候选基因3个:SmCKX(Eggplant_newGene_1802)、SmSTS(Sme2.5_09750.1_g00002)和SmFAR(Eggplant_newGene_4241)。SmCKX是一个茄子细胞分裂素氧化酶/脱氢酶相关基因,与番茄的Solyc04g080820.2和Solyc04g080810.3,拟南芥的AtCKX6具有同源性。SmSTS是茄子水苏糖合成酶相关的基因,与番茄的Solyc01g079300.3基因和拟南芥的AT4G01970.1基因同源性较高。SmFAR是一个脂肪酰辅酶A还原酶相关基因,与番茄Solyc11g067180.2基因(FARx相关基因)同源。以上同源基因参与了植物角质,小檗碱和蜡生物合成途径、细胞分裂素合成途径和水苏糖代谢途径,因此,茄子萼片刺的形成可能具有相同的代谢调控途径。

To explore genes involved in prickle development in eggplant calyx and provide the basic knowledge for the mechanism and gene mapping of prickle development in eggplant calyx,two inbred lines,of which 1607 is the line with prickles while 1608 is not,were used to construct the F 2 population.The two parental lines and two separate pools were then used for RNA sequencing.A total of 409 363 358 clean reads were obtained,of which 311 756 616 were aligned to the eggplant reference genome,with the alignment efficiency over 75.00%.A total of 24 310 new transcripts were found and 1 540 of them were annotated.2 438 genes were differentially expressed between the two parental lines and the two pools,of which 97 were also shared between the two pools.Based on gene annotation,three genes might be involved in the formation of prickle:SmCKX(Eggplant_newGene_1802),SmSTS(Sme2.5_09750.1_g00002)and SmFAR(Eggplant_newGene_4241).SmCKX is acytokinin oxidase/dehydrogenase,SmSTS is a stachyose synthase,and SmFAR is a fatty acyl-CoA reductase.Their homologues function in cytokinin synthesis,stachyose metabolism and wax biosynthesis pathways,individually,indicated that the development of prickle in eggplant calyx might be also affected by the aforementioned signaling.