Transcriptome Sequencing Of Fagopyrum Hailuogouense Rhizome From Dormancy And Growth Stages

Fagopyrum hailuogouense J.R.Shao&Y.Tang,a new wild buckwheat species discovered by our group recently,has the characteristics of perennial,self-pollination and strong stress resistance.It has two reproductive modes,sexual reproduction and vegetative reproduction.Based on the above characteristics,Fagopyrum hailuogouense could be treated as an ideal material for buckwheat breeding research in the future.In order to explore the reproductive regulation network of this species,the Illumina HiSeq^TM 2000 was used for the high-throughput sequencing to producing the transcriptome of its rhizomes from dormancy and growth stages.After that,the key genes related to dormancy breaking and rhizome growth were screened by bioinformatics.Finally,our study revealed the reproductive regulation mechanism of F.hailuogouense,and it also provided a theoretical reference for the breeding of perennial cultivars in other crops.The main results are as follows:1.Morphological observation showed that the dormancy stage of F.hailuogouense began at the end of October under natural conditions.During this period,the aboveground part of F.hailuogouense began to wither,and it could last for half year.On the other hand,the germination and growth of its rhizome began at the middle of April,which happened after the ice and snow melting.At the same time,it was found that F.hailuogouense could reproduce through both sexually and asexually after domestication and cultivation experiments of wild plants.2.The transcriptome sequencing of F.hailuogouense at dormancy and growth stages was carried out for the first time.A total of 48.39 Gb of clean bases were obtained.Through assembly and splicing of Trinity software,289,634 Unigenes were processed.Compared with public databases,98,103,80,081,155,729 and 156,787 Unigenes were annotated in NT,KO,SwissProt and PFAM databases,respectively.A total of 156,787 Unigenes were obtained after GO analysis.It was found that the Unigenes were mainly concentrated in cells and cell components,which might be related to rhizome enlargement.63,505 Unigenes were obtained from NR database annotations.Homology analysis of Unigenes showed that the highest homology was found in Quercus suber Bl.with 557 Unigenes.3.A total of 125,379 DEGs were obtained through screening for genes differentially expressed between dormancy and growth,of which 26,786 Unigenes were down-regulated and 98,593 Unigenes were up-regulated.GO enrichment results showed that GO was mainly concentrated in metabolic components and glycometabolic components.KEGG pathway enrichment results showed that there were 483 Unigenes differential involved in glycometabolism,432 of which were up-regulated and 331 hormone-related Unigenes were up-regulated,121 of which were up-regulated.Among the selected regulatory factors related to rhizome enlargement,there were 4 differentially expressed DELLA-like and 3 up-regulated expression factors;78 differentially expressed MADs-box and 61 up-regulated expression factors;the differentially expressed MYB and WRKY were significantly enriched and showed a complex trend of change.4.11 Unigenes DEGs related to the enlarged rhizome were selected,and the transcriptome data were verified by qRT-PCR.The results showed that the transcriptome sequencing results were basically the same as the transcriptome sequencing results,indicating that the transcriptome sequencing results could clearly reflect the changes of genes in two periods of F.hailuogouense.