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Tissue Culture and Quality-related Gene Cloning in Carthmus Tinctorius L.

Author: ZhuZhenXian
Tutor: PanHuiXin
School: Nanjing Forestry University
Course: Tree Genetics and Breeding
Keywords: Safflower HSYA A (HSYA) Tissue culture Clone Bioinformatic analysis Real-time quantitative
CLC: TQ461
Type: Master's thesis
Year: 2011
Downloads: 48
Quote: 1
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Abstract


Dried tubular flowers of safflower (Carthami Flos) for the Compositae safflower (Carthmus tinctorius L.) , has anti-inflammatory analgesic , expansion of coronary artery , lowering blood pressure and a variety of pharmacological effects , and a wide range of clinical applications . Which the flavonoids HSYA A (HSYA) is the main active ingredient of safflower , whereby the HSYA A content directly affects the the safflower quality the pros and cons of . Therefore , from synthesis related genes the gene level HSYA biological the directional regulation safflower quality , and ultimately from the molecular level to provide a great convenience for safflower breeding . In addition , the use of tissue culture techniques to cultivate the safflower renewable plant regeneration , and to lay the foundation for the rapid propagation of safflower and verify gene function . The main results obtained in this study are as follows : screened safflower seed 0.1% HgCl2 disinfection time : 30 minutes ; induction of the different parts of the explants ( cotyledon , root , stem , leaf ) informed regeneration system to establish the optimal induction material : cotyledons ; trial comparing cell auxin NAA and cytokinin 6- BA , KT , of TDZ different ratio of medium screened most conducive to callus induction and induced the formation of regeneration bud medium : , MS 0.1mg / L NAA 2.0mg/L6-BA 15mg / LKT's and MS 0.1mg / L NAA 2.0mg/L6-BA 20mg/LKT . By high performance liquid technology to complete the determination of the content of safflower in HSYA 2009 , ranging from 0% -2.6 % .3 content is divided into rapid amplification of cDNA ends (Rapid amplification of cDNA ends, RACE) technology flavanones, 3 - hydroxylase (F3H) gene was first cloned from the safflower plant . And the genes of the prediction of the protein structure and preliminary bioinformatic analysis , F3H gene cDNA was 1415bp ( GenBank Accession No. : JF737995 ) contains the a 1086bp open reading frame ( ORF ) encoding 361 amino acid residues . Using real - time quantitative PCR to detect the F3H gene safflower sample HSYA content expression differences . Found that in the Baihua (excluding HSYA ) in the F3H expression far lower than the expression in the yellow (containing HSYA) .

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