Cloning And Expression Analysis Of Genes Related To The 2-phenethylalcohol Biosynthesis In Rosa Rugosa

Rosa rugosa Thunb. is a kind of troditinol famous ornament flower in China and various in varity,full of ornament. Rosa rugosa is not only a good landscaping material,but also an old anatural flavor plant which is rare material.2-phenethylalcohol is a natural spice substance,and it is also the indispensable ingredient in the flavor of rose petal and the essential oil.Taking’Tang hong’which is the leading cultivar of oil-bearing rose as experimental material,we cloned two genes RrAADC and RrPAR which related to 2-phenethylalcohol biosynthetic pathway,and then analyzed the expression of the two genes RrAADC and RrPAR in different developmental stages and different organs of R. rugosa.The relationship between the accumulation of 2-phenethylalcohol and the genes expression in R. rugosa were analyed too.The main results are as follows:1、The full opening petal of ’Tang hong’was used to extract total RNA by the CTAB method,then the full cDNA sequence of RrAADC and RrPAR gene which related to 2-phenethylalcohol biosynthetic pathway were cloned by RACE method.The results have shown that the full length of RrAADC is 1719bp with the start codon, the complete open reading frame（1527bp）,termination codon, the 5’non-conding region（114bp）, the 3’non-coding region （67bp） and poly（A） tail（12bp）,which encodes 509 ammo acid,and the Genbank No. is KJ027532; the full length of RrPAR is 1258bp with the start codon,the complete open reading frame（966bp）,termination codon,the 5’non-conding region（54bp）,the 3’non-coding region （266bp） and poly（A） tail（12bp）,which encodes 322 amino acid,and the Genbank No. is KF700273.By comparing the cDNA sequences with publicly availably reference sequences in Genbank, RrAADC shares 99% homology with Rosa x damascena（BAF6443.1） and 98% homology with Rosa hybrid cultivar Hoh-Jun（AB305011.1）.The homology similarity of RrAADC deduced amino acid sequences and Rosa x damascena （BAF64843.1）, Rosa hybrid cultivar Hoh-Jun（BAF 64844.1） decuced amino acid sequences are as high as 98%,97%; RrPAR shares 98% homology with Rosa x damascena（AB426519.2）,78% homology with Solanum lycopersicuml （NM₀01247894.1）,74% homology with Solanum lycopersicum2 （NM001247901. 1）.The homology similarity of RrPAR deduced amino acid sequences and Rosa x damascena （BAG13450.2）, Solanum lycopersicuml （NP₀01234823.1）, Solanum lycopersicum2 （NP₀01234830.1）,decuced amino acid sequences are as high as 99%,77%,75%.2、We used’Tang hong’as plant material to study the expression patterns of RrAADC and RrPAR in defferent developmental stages and different organs of R. rugosa by RT-PCR and Realtime-PCR,the relationship of 2-phenethylalcohol accumulation and the genes expression were analyed also.The results have shown that the level of 2-phenethylalcohol was increasing with the flower blooming, stongest at senescence stage.The expression level of RrAADC was decreasing all the stage of flower blooming, and the expression level was significantly low at bloom and senescence stage,almost nothing at the two stages; the expression level of RrPAR was increasing from bud to half opening stage, strongest at half opening stage, after that the expression level rapid decresed at bloom stage and then it increased slightly at senescence stage.The level of 2-phenethylalcohol was highest in bloom petals, following was stamens and pistil,and nothing in other parts. The expression level of RrAADC was highest in pistil following is stamens. To compare with the two parts, the expression level of RrAADC was lower in petal, pedicel, calyx, receptacle. The expression level of RrPAR was highest in stamens,next is calyx,while the level was about half in pistil and receptacle than in stamens, and the level was lower in petal and pedicel. So RrAADC and RrPAR play an important role in the biosynthesis of 2-phenethylalcohol in R. rugosa.