基于RNA-Seq技术分析在金属矿上部生长的芒萁的差异表达基因

摘要

以生长于广西大厂锡多金属矿上部(重金属胁迫区)和未受矿化或污染影响的矿区外围(对照区)的芒萁〔Dicranopteris pedata(Houtt.)Nakaike〕为实验材料,对芒萁叶片进行转录组高通量测序,并对组装得到的unigenes经NCBI官方非冗余蛋白质序列数据库(Nr)、NCBI官方非冗余核苷酸序列数据库(Nt)、KEGG直系同源数据库(KO)、Swiss-Prot数据库(Swiss-Prot)、蛋白质家族数据库(Pfam)、基因功能分类体系数据库(GO)和真核生物直系同源序列数据库(KOG)进行注释,同时分析重金属胁迫区和对照区芒萁叶片间的差异表达unigenes.结果显示:测序获得19.56 Gb clean data,其中,重金属胁迫区和对照区芒萁叶片分别含10.14和9.42 Gb clean data.组装得到的250582个unigenes中有120097个unigenes得到注释,占unigenes总数的47.93%.与对照区相比较,重金属胁迫区芒萁叶片中上调和下调差异表达unigenes分别有208和620个,其中120个上调差异表达unigenes注释为代谢过程,占所有上调差异表达unigenes的57.69%;285个下调差异表达unigenes注释为催化活性,占所有下调差异表达unigenes的45.97%.重金属胁迫区芒萁叶片中15个unigenes与重金属转运和耐受相关,其中c44988 g1和c84121 g1的相对表达量分别极显著和显著高于对照区.研究结果显示:芒萁响应自然金属矿化或矿山重金属污染的基因可以用于生物地球化学找矿和土壤重金属污染检测.%Taking Dicranopteris pedata (Houtt.) Nakaike grown on tin polymetallic deposits (heavy metal stress area) in Dachang of Guangxi and on the periphery of mining area without mineralization or pollution ( the control area ) as experimental materials, the transcriptome high-throughput sequencing was conducted on the leaves of D. pedata, and assembled unigenes were descripted by NCBI non-redundant protein sequence database ( Nr ) , NCBI non-redundant nucleotide sequence database ( Nt ) , KEGG Orthology database (KO), Swiss-Prot database (Swiss-Prot), Protein family database (Pfam), Gene Ontology database ( GO) , and euKaryotic Orthology Groups database ( KOG) . Meanwhile, differentially expressed unigenes between leaves of D. pedata in heavy metal stress area and the control area were analyzed. The results show that 19. 56 Gb clean data are acquired by sequencing, in which, leaves of D.pedata in heavy metal stress area and the control area include 10. 14 and 9. 42 Gb clean data, respectively. 120097 unigenes from 250582 assembled unigenes are descripted, with 47. 93% of total number of unigenes. Compared with the control area, there are 208 and 620 up- and down-regulated differentially expressed unigenes in leaves of D. pedata in heavy metal stress area, respectively. In which, 120 up-regulated differentially expressed unigenes are descripted in metabolism process, accounting for 57. 69% of all up-regulated differentially expressed unigenes, and 285 down-regulated differentially expressed unigenes are descripted in catalytic activity, accounting for 45. 97% of all down-regulated differentially expressed unigenes. Fifteen unigenes in leaves of D. pedata in heavy metal stress area are associated with heavy metal transport and tolerance, in which relative expression of c44988 g1 and c84121 g1 are extremely significantly and significantly higher than those in the control area, respectively. It is suggested that genes in D. pedata in response to natural metal mineralization or heavy metal pollution of mine could be used for biogeochemical prospecting and soil heavy metal pollution detection.