Transcriptome Analysis of Highly Efficient Rooting in Mulberry Softwood Cuttage and Validation on the Related Genes

Mulberry is the material basis of sericulture. Cuttage is one of the importantseeding methods, as well as a economical-effective method for other woody plant’s.On the basis of the efficient artificially induced rooting, we transform the study anglefrom traditional cellular level, for example morphological, anatomical, physiologicaland biochemical indexes et al., to go deep into molecular level. We can comprehendthe mechanism of mulberry softwood cuttings rooting more accurate by using thetranscriptome sequencing technology, and of course it could conductive to the furtherinnovation of cutting technology.Solexa sequencing technology has been developed rapidly in recent years that is ahigh-throughput sequencing technology. Comparing to the traditional sequencing,high throughput sequencing has with a wide degree of information coverage and adeep sequencing feature, low data redundancy as well as analysis accurate of newspecies transcript expression profile with the unknown genomics background.In this paper, using Solxea sequencing and bioinformatics analysis technology tostudy the transcriptome of mulberryvarieties Yu71-1cutting rooting, and the results asfollow:1. A total of more than230M raw reads was obtained from the three samples thatthe different development course. We obtained88,216,727、88,616,727and52,756,096high-quality reads respectively after the format conversion and removal ofimpurities sequence, and61687、61651and59457contigs after assembling.2. The contigs protein annotation in uniprot database and functional annotation:Gene Ontology (GO) function annotation; Kyoto Encyclopedia of Genes andGenomes (KEGG) function annotation and Simple Sequence Repeat (SSR) annotation.The protein annotation results of the three libraries was that28042、32755and30587contigs were annotation with the annotation rate45.46%,53.13%and51.44%.3. A wide range of gene regulation about adventitious formation were found bypairwise comparison of the data among the different samples.231different expressedgenes between day0and day4, with148up-regulated and83down-regulated;4183different expressed genes between day4and day8, with1831up-regulated and2352 down-regulated;4557different expressed genes between day0and day8, with1930up-regulated and2627down-regulated.4. GO and KEGG analysis of the differentially expressed genes during thecuttings adventitious formation course, some significant difference pathways werefound, such as Glycolysis/Gluconeogenesis, RNA transport, Plant hormone signaltransduction and Starch and sucrose metabolism et al.. Also some mulberry cuttingsrooting related genes were found (such as lpi, HSP70, cTBP, CHS, ECPP44, M35andso on).5. We detected relative expression level of the two ethylene-related-genes acoand sams during the development of mulberry softwood cutting rooting by real-timequantitative PCR. The results illustrate that the two genes aco and sams may play anegative correlation effect on the mulberry softwood efficient rooting.