Investigation Of Cultivars Of Bougainvillea And Expression Analysis Of Floral Related Genes

Bougainvillea(Bougainvillea spectabilis Willd)has a high ornamental value,often used in landscape gardening and afforestation plant,is the city flower of xiamen,this paper adopts literature index,visiting survey,typical methods of interview and evaluation research,of Bougainvillea different colors make a preliminary study of the ornamental;And from the transcription and biochemical level,clarify the bougainvillea bracts color formation,the molecular mechanisms of flowering,color and for bougainvillea bracts flowering gene regulation and the development and utilization of color,have important theoretical and practical significance. The main results were as follows:1.Using the analytic hierarchy process(ahp)to establish bougainvillea varieties of ornamental characters and comprehensive evaluation system:the comprehensive evaluation score of the bougainvillea cultivars is divided into four grades(Ⅰ, Ⅱ,Ⅲ,Ⅳ),its ornamental value sequence in the order:Bougainvillea peruviana ’Imperial Delight’>Bougainvillea × buttiana ’Tetra Mrs.Mcclean’> Bougainvillea peruviana ’Mrs Butt’> Bougainvillea ’Bottle Brush Orange’> Bougainvillea ’Double Delight’> Bougainvillea ’Scarlet Queen Variegated’> Bougainvillea ’Fatima’> Bougainvillea × buttiana ’Golden Glow’> Bougainvillea ’Drange Stripe’> Bougainvillea ’Cindrella’> Bougainvillea glabra ’Pink Pixie’> Bougainvillea ’Badgen’s Beauty’>Bougainvillea ’Elizabethh Angus’> Bougainvillea glabra ’Dr.H.B.Singh’> Bougainvillea × buttiana ’Red Dania’> Bougainvillea ’glabra Formosa’> Bougainvillea ’Sweet Dreams’> Bougainvillea sp .’Begum Sikander’> Bougainvillea ’Coconutlce’> Bougainvillea × buttiana ’suerkha’> Bougainvillea glabra ’White Stripe’.Foloral colors,leaf color,flower quantity is still the main factors influencing the bougainvillea ornamental value. ’Imperial Delight’,’Tetra Mrs.Mcclean’with Varieties for rare degree is higher.with a strong growth potential,’Mrs Butt’Due to the average value is higher rank the first three.2.Build the bougainvillea bracts full-length homogenization cDNA Iibraries:This research took’Imperial Delight’at Bracts as material,Reverse transcribed into cDNA by two-step amended SMART cDNA synthesized method which has higher proportion of full-length cDNA.The titer of primary library was about 9.0×106pfu/ml and 97%clones were recombinant,and the length of insert cDNAs were about 1.5 kb.and database is 5.0×107.Meet the requirements of sequencing and the analysis of EST.By EST analysis,and find out the formation and bougainvillea bracts color gene flavanone-3-hydroxylase(F3H),containing two by MADS box-flowering gene(AP1 and AG6),photoperiod induce flowering with zinc finger protein gene(COL).3.Bioinformatics analysis and expression analysis of Bougainvillea bracts F3H,AP1,AG6 and COL:Bioinformatics analysis of the sequence of amino acids deduced by this gene showed that bougainvillea bracts F3H cDNA which has a full sequence of 1330 bps containing 1146 bps open read frame,a 6 bps 5’UTR and a 164 bps 3’UTR,respectively,encoding 381 amino acids with contains DIOX_N and two 20 g-FeⅡ_Oxy Structure of the domain,the BLAST analysis results show that The gene on the amino acid sequence homology with sesame and citrus were 59% and 59% respectively,can be presumed this gene for bougainvillea F3H homologous genes;Fluorescence quantitative PCR results showed the gene in the different stages of bougainvillea bracts,its expression level is different,in the early days of bracts development.F3H in expression,expression quantity at a higher level,after the first decline after the rising trend,illustrate F3H expression is associated with the colour and lustre of bougainvillea bracts formation.After sequencing,the full length of Bougainvillea bracts API cDNA which has a full sequence of 1247 bps containing 743 bps open read frame,a 226 bps 5’UTR and a 253 bps 3’UTR,respectively,encoding 247 amino acids,having MADS-box and two K-box conserved domains.BLAST analysis showed that the gene in amino acid sequence has high homology with spinach,tea and grapes,respectively 82%,75% and 74%.It can be presumed that the gene is the homologous gene of Bougainvillea AP1;and the fluorescent quantitative PCR results showed that the gene is expressed at different times of Bougainvillea bracts,in the early development of the bracts,it has strongly expression quantity,but the expression quantity is gradually decreased along with the growth of bracts.Sequencing analysis showed that the full length of Bougainvillea bracts AG6 cDNA which has a full sequence of 1124 bps containing 801 bps open read frame,a 29 bps 5’UTR and a 228 bps 3’UTR,respectively,encoding 267 amino acids,having MADS-box and the two K-box conserved domains.BLAST analysis showed that the gene in amino acid sequence has 72% homology with beet.It can be initially speculated that the gene is the homologous gene of Bougainvillea AG6;and the fluorescent quantitative PCR results showed that the gene expresses differences at the different developmental stages of the Bougainvillea bracts formed.In the early development of bracts,it has the highest expression quantity,while the amount of expression appears to reduce along with the growth and turn color of the bracts.But its expression quantity appears to rise after bracts stop growing.Since the Bougainvillea API and AG6 which isolate by this study have conserved domains of flowering genes(MADS-box and the K-box),and in the early development of bracts,both have the high expression quantity,then appears to decline,therefore,these two genes are likely closely related Bougainvillea flower.Sequencing results showed that the full length of Bougainvillea bracts COL cDNA which has a full sequence of 1364 bps containing 1090 bps open read frame.a 95 bps 5’UTR and a 259 bps 3’UTR,respectively,encoding 364 amino acids,there are respectively a CCT domain and two K-box conserved domains in its N and C-terminal.It is BBX protein.BLAST results showed that the gene in amino acid sequence has 74% homology with beet.It can be initially speculated that the gene is the homologous gene of Bougainvillea COL,and being related with photoperiodic induction of flowering;fluorescence quantitative PCR results showed that the gene was expressed in different developmental stages of Bougainvillea bracts.In the early development of the bracts,it has strongly expression quantity,but the expression level difference is not very big.