Cloning Of Genes For Partial Member Enzymes Of Carotenoid Biosynthetic Pathway In Strawberry (Fragaria Ananassa Duch)

Strawberry (Fragaria ananassa Duch) is a health-care fruit of high economic value, and is a top grade fruit. Carotenoids is not only an important indicator to decide intrinsic nutritional quality of fruits and vegetables, but also an important factor which affected the appearance quality of fruits and omamental value of flowers. Comprehending the accumulation and metabolism physiological mechanism of plant carotenoid, it is of important to improve the eating, viewing and admiring quality of horticultural plants, and to control and regulate the further biosynthetic pathway of plant carotenoids. Strawberries are rich in carotenoids, but current study of Strawberry is less concerned about carotenoids. The separation and identification of strawberry carotenoid biosynthetic pathway-related genes, the structure and function of various metabolic enzymes, as well as the accumulation and metabolic mechanism of carotenoids had not been reported at home and abroad.'Fengxiang'Strawberry fruit was selected as test materials in the project, key gene PSY, ZDS and PDS which determined the pathway carotenoid synthesis of strawberry were cloned, the foundation for mechanism of Strawberry carotenoid synthesis was lay, a wealth sources of the strawberry breeding gene was provided for further use of genetic engineering to improve the quality of strawberries and other crops.The project had wide economic value and social value. The main test results were as follows:1.The extraction method of strawberry fruit RNA samples without the pollution of protein and polysaccharide, and with more complete, no degradating RNA, was established.2.Full-length cDNA sequences of Strawberry PSY gene was cloned, the complete cDNA sequences cotains 1458 bp, with a complete open reading frame of 1194 base pairs, encoding 398 amino acids. The consistency between deduced amino acid sequence of the complete sequence of the open reading frame and the amino acid sequences of maize (NP001108117.1), carrot (Q9SSU8.1), marigold (AAM45379.1), papaya (ABG72805.1), pomelo (ABY86652.1) were 80%, 75%, 72%, 74%, respectively, and the similarity were 87%, 85%, 81%, 84%, respectively. The plant PSY protein system evolution was analysis by DNAMAN software, discovering that the PSY protein relationship between strawberry, carrot and corn were close.3.The full-length cDNA of Strawberry PDS gene was cloned, the complete cDNA sequences cotains 2043 bp, with a complete open reading frame of 1704 base pairs, encoding 568 amino acids. The consistency between the complete sequence of the open reading frame deduced amino acid sequence and apricot (AAX33347.1), grapefruit (AAK51545.1), pomelo (ACE79168.1), papaya (ABG72807.2) were 87 %, 86%, 86%, 81%, respectively, and the similarity were 92%, 93%, 93%, 88%, respectively. The plant PDS protein system evolution was analysis by DNAMAN software, discovering that With the PDS protein relationship between strawberry and apricot protein were relatively closer.4.The conservative region of strawberry ZDS gene was cloned and with 3'end of 1709 bp of sequence in total. The 3'end fragment encode 488 amino acid residues, contain a non-coding region of 242 bp. Blasting by the NCBI Web site search, the consistency of the amino acid sequences between Apple (AAQ04224.1), citrus (BAB68552.1), pomelo (ACE79169.1), sweet orange (CAC85667.1) were 92%, 89%, 88%, 88%, respectively, and the similarity were 96%, 94%, 93%, 94%, respectively.