Study Of The Seeds Primary Substances Growth And Clone Of AccD Gene From Aleurites Moluccana

Aleurites moluccana belongs to woody-oil plants in Euphorbiaceae and is rich in oil in its seeds. It is also a sort of extremely important biomass energy plant. Firstly, comparative analysis on substances of seeds of the different plants in Aleurites moluccana were conducted, and content changes in the amount of soluble sugar, soluble protein and oil fat were tested in Aleurites moluccana in different fruit growth stages. Secondly, to clone full length cDNA sequence of accD gene of acetyl coenzyme A carboxylase (ACCase)β-CT subunit from tender leaves of Aleurites moluccana and analyze its sequence is to provide referential data for the research of using this kind of gene and increasing the oil content of oil-bearing crops seeds. Last, Tender leaves of Aleurites moluccana was used as the material, and RT-PCR technology was used to clone the actin gene, which can be used as internal standard when studying other genes’ expression and regulation from leaves of Aleurites moluccana. The main results in the paper are as follows:1、Study of the seeds substances changes in Aleurites moluccanaResults show that there are varying degrees of differences in the fruit seeds rate、water content、oil fat and protein of seeds of the different plants in Aleurites moluccana, and there exists correlation between the level of water content and fruit of seeds rate, oil fat and protein. The content percentage of oil fat of different Aleurites moluccana plants is about62%to68%. The content percentage of protein of different Aleurites moluccana plants is about15%to23%.With the seeds development during the fruit growth, in the peak of the soluble protein is210.32mg/g after flowering180days, in the low of the soluble protein is66.34mg/g after flowering75days; The soluble sugar content becomes the lowest at the lateral stage of fruit development, which is32.05mg/g after flowering180days, in the peak of the soluble sugar is99.52mg/g after flowering105days; In the peak of the oil fat is68.33%after flowering180days, in the low of the oil fat is8.15%after flowering75days.2、Cloning and analysis of full length cDNA sequence of accD gene from Aleurites moluccanaDegenerate primers were designed based on the conserved region of known accD gene of plants, such as Jatropha curcas and Ricinus communis. Tender leaves of Aleurites moluccana were used as the material for extracting total DNA and RNA, PCR and RACE technology were used to clone and amplify the full length cDNA sequence of accD gene. We obtained an1789bp sequence of full length cDNA of accD gene, which contained1509bp open reading frame and could encode a503amino acids protein. The accession number of GenBank is KC255250, which was named as amaccD.3、Cloning and sequnence analysis of actin gene fragment from Aleurites moluccanaTender leaves of Aleurites moluccana were used as the material, and RT-PCR technology was used to clone the actin gene. We obtained a396bp nucleotide sequence of actin gene, which could encode an132amino acids protein. The results of GeneBank showed that the degree of identity of the nucleotide acid sequence between actin gene of Aleurites moluccana and that of known homologous plants is over90%, the accession number of GenBank is KC767896, which was named as amactin.