Transcriptome Analysis Of Rooting In Mulberry Cutting And Expression Analysis The Genes MaGH3.6and MaHO-1in Mulberry

In this paper, a genome-wide gene expression profiling was conducted bySolexa sequencing to investigate of the transcriptome dynamics of two typesmulberry cutting rooting. And MaSAUR2and MaHO-1genes, which significantlydifferential expressed in the cutting rooting, were analyzed by cloning, sequencefeature analysis and expression pattern analysis.The main results and conclusion were as follows:1.According to the development of cuttings,3transcriptome database wereestablished by using the summer and autumn greenwood cutting rooting material:W1, W2， W3. And4transcriptome database were established by using the lateautumn and early winter greenwood cutting rooting material: Q1, Q2, Q3, Q4. Weobtained52,756,096to88,216,727high quality contigs after preprocess in eachdatabase. The protein annotation results of the seven libraries were that23837to39504contigs were annotation with the annotation rate35.81%to57.20%.According to the corresponding relationship between contigs and protein annotation,the opposite development stages transcriptome databases of mulberry two typesgreenwood cutting rooting were cross-over analyzed. The W1with Q1，W2with Q2,W2with Q3, W3with Q4databases were get together to analyzed respectively, andwere named A, B, C, D databases.14191uniprot annotation were obtained in A,14142in B,13760in C,12355in D.77different expressed genes were obtainedbetween A and B.1752different expressed genes were obtained between B and C.1991different expressed genes were obtained between C and D. And the differentexpressed genes were clustered by GO and KEGG analysis. Several functionalgroups of genes involved in important pathways were differently expressed,including the energy metabolism, carbonhydrate metabolism, hormone signaltransduction pathway, cell division and differentiate.2.In this study, a SAUR (Small anxin-up RNA) cDNA sequence was clonedfrom stem cortex of Morus alba L. and named MaSAUR2.(GenBank accessionnumber: KC852881). The bioinformatics analysis showed that the gene encodes182amino acids and has a molecular mass of about20.7kDa with an isoelectric point of9.39. The protein has a SAUR-specific domain (SSD). MaSAUR2expressed activelyin root and relatively high in stems and leaves. The expression quantity of this gene is relatively less in the bud, but not expressed in ripe fruit. Expression levels ofMaSAUR2were signigicantly increased within5minutes in the stem cortex ofmulberry cutting after treatment with IAA. In mulberry cutting rooting, MaSAUR2has a lower expression level in the enlargement stage of cutting base, but increasedsignificantly during adventitious tootsie grow out of cortex. Preliminary view is thatthe gene palys an important role in the root elongation of mulberry cutting.3. In this study, a HO (heme oxygenase) cDNA sequence was cloned from stemcortex of Morus alba L. and named MaHO-1.(GenBank accession number:KJ544669). The bioinformatics analysis showed that the gene encodes291aminoacids and has a molecular mass of about26.2kDa with an isoelectric point of8.79.The protein has a Hame oxygenase-like domain. MaHO-1expressed active in leaf,followed by bud, stem, mature fruit, root. Expression levels of MaSAUR2wereincreased gradually in the stem cortex of mulberry cutting after treatment with IAA.In mulberry cutting rooting, MaHO-1maintains a higher expression level in themulberry cutting rooting. Expression quantity increased5times from control to theenlargement stage of cutting base, then increased slow. Change of heme oxygenaseactivity was basically the same with the gene expression. And the expression of geneincreased before the formation of adventitious root. Preliminary view is that thegene palys an important role in the root formation of mulberry cutting.