| Anthurium andraeanum is one of the world's rare flowers with high ornamental value and is the international popular upscale cut flower material.The flower is composed of spathe and spa dix.The spathe is main ornamental parts,it has red,white,orange,pink and green color,but t he lack of yellow,blue color.Spadix with yellow,red,pink,and green color.In general,Anth urium andraeanum flower just has pink and white color,there are little diversities in the comp ose of flower color.Cinnamate-4-hydroxylase(C4H)is the second important pacemaker enzyme i n the benzene propane pathway,it also involves in the catalytic synthesis of a variety of secon dary metabolites.'Alabama' as material in this study,by RT-PCR in combination with RACE te chnology get a cinnamate-4-hydroxylase gene.To study the genetic biological information,to an alyze its expression changes.Select flower pigment function and regulation of genes in the met abolic pathway,research expression patterns under different flower development stages and pH.Proposed for flower color of Anthurium andraeanum metabolic research,as well as the subsequ ent flower colormolecule is improved,and increase the diversity of flower color research provid es the basis.The main results were as follows:1.A C4H gene was isolated from the 'Alabama' spathe in Anthurium andraeanum by RT-PC R and the RACE,it is named as AaC4H.The full-length cDNA of AaC4H was 1813 bp with an open reading frame of 1512 bp.BLASTN showed that AaC4H amino acid sequence with si milarity reaches 76%-87%in Morus alba,Tulipa fosteriana,Ginkgo biloba,etc.2.C4H protein structure prediction analysis showed that:its molecular formula is C2610H4133N721O719S16.C4H protein is composed of twenty kinds of amino acids,protein molecular weight 57629.8 Da.The system evolution analysis shows:AaC4H with monocotyledon plant C4H gene clustering together,it reflects the close genetic relationship between monocotyledon plant C4H gene,3.Expression of RT-PCR semi-quantitative analysis the seven functional genes in anthocyanin biosynthetic pathway at different organizations in 'Alabama',`White sun' and `Madici'.It tune d out that(1)in 'White sun',C4H?CHS?F3H?DFR?ANS and R2/R3MYB six different genes express nearly equal in vegetative organs and former spathe,the others express lower or no ex pression.In 'Madici',the six genes has the lower expression quantity,the former spathe has 1h e highest expression level.In 'Alabama',CHS' F3H' ANS' R2R3MYB express nearly equal in ve getative organs and former spathe,DFR only express in spathe,C4H has the highest expression 1evel in root and stem,unexpress in later spathe and spadix.(2)in 'White sun' and 'Madici',F3'H has no expression,in 'Alabama',the former spathe has the less expression level.So,differ ent cultivars'genes has its expression pattern.The study provide a basis for clarify the color m echanism of different breed of design and color.4.Expression of real-time PCR analysis the functional genes in anthocyanin biosynthetic pathway at different flower developing stages of Alabama.The result shows that C4H?DFR?R2R3MYB appeared a decreasing trend,in stage 1 has the highest expression level,in stage 4,5 has the lowest expression level.CHI?CHS appeared a rising trend,in stage 1 has the lowest expression level,in stage 4,5 has the highest expression level.F3H appeared N type express mode,in stage 3-3 has the highest expression level,stage 5 next.C4H?ANS and MYB1 appeared fluctuation trend.The research provide the basis for the molecular breeding of flamingos colors.5.Expression of real-time PCR analysis the genes in anthocyanin biosynthetic pathway of An thurium andraeanum at different pH.The result shows that in 'Madici,other genes except C4H appeared at 7.0 has the highest expression level.In 'White sun' and 'Alabama',almost genes a t 5.5 or 7.0 has the highest expression level.The study provide a basis for the optimum pH v aluecontrol with different varieties,cultivation flamingos matrix or nutrient solution. |
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