High-throughput Analysis Of Endogenous Ethylene Dynamics And Related Genes In The Floral Organs Of Different Lonicera Japonica Varieties

Lonicera macranthoides Hand.-Mazz.which is from Caprifoliaceae and as one of the main sources of Flos Lonicerae Confusae.In last century,compared with the conventional Lonicera Macranthoides,the natural mutation clone“Xianglei Lonicera Japonic",which is for introduction and cultivation,had been found that it had many good characters,such as a long bud period,unexpanded Corolla,clustered and neat buds,uniform bud color and disposable picking and so on.To explore the reason why Xianglei Lonicera Japonic has the obvious flowering differences,the research obtained the corresponding results from the several following aspects,and laid a solid foundation for the final fundamental reason to flowering differences from the molecular level.The main research topics and results include the following:1?The dynamic changing of the endogenous ethylene producing rates from these two varieties of Lonicera Macranthoides was measured by gas chromatographic.The results showed that during the whole flowering process,the conventional variety endogenous ethylene content was always higher than Xianglei Lonicera Japonic variety's,especially the later flowering period formation rate of the latter had significantly increased.2.The total RNA in buds which are about to open from Lonicera macranthoides Hand.-Mazz.were extracted using five different methods(SDS method,CTAB method,ordinary kit method,polysaccharides polyphenols kit method and modified polysaccharides polyphenol kit method).The UV and agarose gel electrophoresis were used to detect the purity and integrity of RNA.From the results analysis of comprehensive purity and integrity detection,the RNA which were extracted by ordinary kit method,polysaccharides polyphenols kit method and modified polysaccharides polyphenol kit method,can basically meet the follow-up experiments such as cloning,sequencing and so on.Modified polysaccharide polyphenols kit method was the most suitable method of extracting total RNA in high quality from Lonicera macranthoides Hand.-Mazz.buds.3.The high quality extracting total RNA were determined using high flux sequence and sequenced on the platform of Illumina Hiseq 2000.9597839040nt bases are generated totally.In the results of assembly,67222 unigenes are detected,total length for unigenes is 53645869 nt,average length is 798 nt.The unigenes in two varieties' which have obviously different expression quantity are 20709 pieces,in which compared with the conventional Lonicera Macranthoides,the upregulated expression quantity in the natural mutation " Xianglei Lonicera Japonic"are 11779 pieces,and the downregulated expression quantity are 8930 pieces.In these obviously different genes,ACS regulates 6 pieces of family genes and ACO regulates 8 pieces of family genes.Besides,the MADS-box family genes are 30 pieces.