The Identification Of The Acid Invertase Gene Family And Analysis Of Biological Function In Phyllostachys Edulis

In order to study the role of acid invertase gene family in flowering and bamboo shoot developing in Phyllostachys edulis,we obtained the acid invertase gene family from the genomic database of Phyllostachys edulis based on the sequence of rice invertase gene family and analyzed expression pattern by RT-PCR.Then we transferred the PhCWINV1,PhCWINV4 and PhCWINV7 genes into wild type Populus to verify the function by phenotype,and analyzed the intracellular localization of PhCWINV1,PhCWINV4,PhCWINV7 gene.Meanwhile,we also measured enzyme activity,sucrose and glucose content,cellulose content and other indicators in flowering time and bamboo shoot during development in Phyllostachys edulis.The results of the study are as follows:1.Fourteen invertase homolog genes were obtained from genome database of Phyllostachys edulis,named PhCWINV1-10 and PhVINV11-14 respectively.The full length ranges from1962-12577 bp and exon number between 3-10.Nine invertase homolog genes were obtained from the CDS database,and the cDNA length of acid invertase gene ranges from 1029bp-2352 bp,encoding 342 to 783 amino acid respectively.2.In order to study the role of acid invertase gene family in flowering and bamboo shoot developing in Phyllostachys edulis,we detected the expression of gene family.In the semi-quantitative analysis showed that all genes are expressed in at least one organization.PhCWINV1-5,PhCWINV6 and PhCWINV12 are ubiquitous expression,and expression almost in all examined tissues.While the spatiotemporal expression of PhCWINV6 and PhVINV11 are more obvious.The expression of PhCWINV1 and PhCWINV2 are similar at leaf,flower buds and the beginning development of bamboo shoots,and the expression is higher than other tissues,but the expression of PhCWINV2 weaker than the expression PhCWINV1;The expression of PhCWINV3 in the upper part of 300 cm is higher,there is almost no change in remaining parts;The expression level of PhCWINV4 is higher in leaf and the base of development shoots;PhCWINV5 not expressed at the base of 150 cm,and there is little change of expression level in other organizations;PhCWINV6 in flower bud is highly expressed,but only weak or no expression in other tissues;The expression of PhCWINV7 are higher in flowering and shoots during the rapid development;PhVINV11 mainly overexpressed in rapid growth phase(150cm);PhVINV12 is expression highly in the upper part of 40 cm and the middle of 150 cm,the expression of the remaining parts are moreconsistent.3.In order to study the role of the enzyme activity in different developmental processes,we measured the enzyme activity,sucrose and glucose content during developmental stage and flowering time.The results indicate that the enzyme activity is highest in middle which is fastest growing at 40 cm and 150 cm,while the activity is highest in the top at 300 cm.The enzyme activity is higher in flower buds than leaf.Glucose content has a substantially similar trend with the enzymatic activity and the sucrose content has a roughly opposite trend with the enzyme activity.4.In order to study the differences in cellulose and lignin content in different parts of bamboo shoots,we measured the cellulose and lignin content from three parts of bamboo shoots which growth stably.The results indicate that the celluose content was the highest at the base,and the top was the lowest,but the difference was not significant.There is a same trend between lignin content with cellulose content,but the the difference is significant.5.For study the differences of inter-fiber length and internode length in different parts of the annual bamboo,the fiber length and the internode length was measured in three parts.The results indicate that there is a similar trends between the fiber length and internode length,the middle is the longest,followed by the base,the top is the minimum,and the difference of three parts is significant.6.By transgenic technology,PhCWINV1,PhCWINV4 and PhCWINV7 were transferred into wild Populus to verify the function by phenotype.7.We constructed PhCWINV1,PhCWINV4,PhCWINV7-YFP fusion plasmid,then imported them in tobacco,Observing the fluorescence signal under a fluorescence microscope.The intracellular localization of PhCWINV1,PhCWINV4 and PhCWINV7 were analyzed.The result showed that PhCWINV1,PhCWINV4 and PhCWINV7 localized in the cell wall.