Transcriptomic Analysis Of Response To Alternaria Sp.infection In Chrysanthemum Morifolium Ramat.Mainly Produced In Ancient Huaiqing Area

Chrysanthemum morifolium Ramat.is one of the famous "four huaiqing Chinese medicines" in our country and has a huge commercial value.A major production and quality constraint on chrysanthemum is black spot which is caused by the necrotrophic fungus Alternaria sp.Planting resistant varieties can effectively control the occurrence and popular of the disease.At present,breeding technology of cultivating chrysanthemum disease-resistant varieties from the molecular level is safe and reliable,But the molecular basis of the chrysanthemum resistance to A.sp.remains to be further studied.This experiment extract total RNA of chrysanthemum leaf which were vaccinated sterile water in 0 d and A.sp in the 3 d and 5 d,construct the cDNA library of the sample,and using the Illumina HiSeq 2000 sequencing platform have on transcriptome sequencing analysis,the results are as follows:1.Won the basic information of the transcriptomic response of the chrysanthemum leaf to A.sp.inoculation.The sequencing received a total of 544411736 Raw Reads.After filtering,Cm_CK0₁,Cm_CK0₂,Cm_CK0₃,Cm_A3₁,Cm_A3₂,Cm_A3₃,Cm_A5₁,Cm_A5₂,Cm_A5₃ respectively won the 56583406,62933114,53182862,62933114,47401094,62287656,66060304,58149022,57499778 high quality clean Reads,a total of 58.6 G.Recognition accuracy up to 99% of the bases in each sample sequence were over 94%.After assembly,take the longest transcription as Unigene,got 350833 transcript and 162252 unigene,their average length is 822 bp and 701 bp respectively,the shortest 201 bp,the longest 106811 bp,medium length is 537 bp and 397 bp respectively,the length of transcript and unigene above 500 bp have 185500 and 64500 respectively,accounted for 52.9% and 39.8%,and above 1000 bp have 92614 and 30545 respectively,accounted for 26.4% and 18.8%.The proportion of the short sequences is more than a long sequence,which accords with the general law of distribution of the length of the transcriptome.2.Get the unigene functional annotation information in multiple databases.By Blast matching Unigene sequences and protein database NR,GO,KOG,KEGG,SwissProt,PFAM and nucleic acid database NT,E-value of NR,NT,SwissProt and KOG is 1e-5,GO for 1e-6,KEGG for 1e-10.A total of 57890 unigene got functional annotation in at least one database,accounted for 35.67%,5124 unigene got functional annotation in 7 database at the same time,48795 unigene got functional annotation in NR protein database,17796 unigene got functional annotation in NT nucleic acid database.3.Got GO,KOG and KEGG function classification information of unigene.Based on annotation information of Nr and Pfam two databases,three categories are classified to GO function classification of 37239?23%?of Unigene involved to 47 terms.Three categories of biological process,molecular function and cellular components respectively 89121?54.9%?,55661?34.3%?and 45263?27.9%?unigene;binding and cellular process in 47 terms contains the most number of unigene,there were respectively 21179?56.9%?and 20485?55.0%?.16034 unigene in KOG database is commented on the 25 functional category,including the largest number general function prediction only contains 3037 unigene,accounting for 19.0%.In the Unigene three-level classification,KEGG pathway is divided into five major branches in the level of classification,and five major branches is divided into 32 pathway,which the Unigene number of signal transduction is the most,including 1673,accounting for 11.7%.32 pathway in tertiary classification is divided into 276 pathway,which the Unigene number of carbon metabolism is the most,including 573,accounting for 3.47%.In addition,such as pathways associated with disease resistance of plant hormone signal transduction and plant-pathogen interaction,etc have 381 and 363 unigene respectively.4.A preliminary understanding of the unigene structure information.Analysis CDS and SSR of unigene,won the 85218 CDS sequence of unigene and distribution of 14705 SSR loci in 14705 unigene.CDS length distribution of Unigene in 201-300 bp is the most numerous and mono-nucleotide repeat in SSR types is the most numerous.5.Has been clear about the basic information of differentially expressed genes.?1?In Cm_A3 vs Cm_CK0 and Cm_A5 vs Cm_CK0,DEGs of |log2foldchange|>l have respectively16550 and 13559,including the same 8422 DEGs;?2?Protein phosphorylation and protein kinase activity is the most significant in GO function classification,KEGG significant enrichment in the plant-pathogen interaction,plant hormone signal transduction,phenylpropanoid biosynthesis,flavonoid biosynthesis and so on pathways,with the most significant of plant-pathogen interactions and plant hormone signal transduction.these functiona category and pathways are related to plant resistance and up-regulation differentially expressed genes is more than down.In addition,the pathways of those involved in photosynthesis and other related some of the differentially expressed genes were significantly lower.?3?Chrysanthemum under the stress of black rot,launched multiple signaling pathways,including by SA,JA/Eth,MAPK and Ca²⁺ mediated,probably by regulating the balance of active oxygen to resist the invasion of pathogenic bacteria,activate the WRKY,ERF,MYB,NAC,bHLH and so on lots of high expression transcription factors related to disease resistance,MPK6 / MPK3 way and WRKY33 transcription factors may play a key role.Above transcript to reply chrysanthemum black rot pathogen and its potential function analysis,enriches the gene expression patterns,a large number of candidate differentially expressed genes are discovery,and for the cultivation and research of chrysanthemum disease-resistant varieties and the mechanism of resistance to black rot laid a foundation.