Five Black Grain Wheat Lines Cytogenetic Analysis And Relevant Pigment Gene Cloning

Black grain wheat is packed with protein, vitamins,minerals, biological brass, amino acids, octacosanol, dietary fiber, anthocyanins and varieties of nutrients. Caryopses in black grain wheat are rich in anthocyanin, which leads to this precious germplasm resource, black grain wheat has significant hygienical function in such diseases as anti-tumor, anti-inflammatory, anti-allergic, hypoglycemic, scavenging free radicals, anti-aging, anti-radiation, immunity enhancement and prevention of cardiovascular.In this study, morphological and cytological observation, Molecular markers and GISH were used for the cytogenetic analysis of the five black grain wheat lines; and carrying on research on the cloning, bioinformatics analysis and gene expression analysis of F3 H which is the related genes of anthocyanin biosynthetic pathway, so as to study the source of black grain wheat anthocyanin and the mode of synthesis metabolism regulation, which provides fundamental basis for the health care functionality of wheat.The result of morphology and cytology shows that: ‘Hashi3-37-1’, ‘Hashi3-39-1’, ‘Hashi3-13-1’, ‘Hashi10-38-1’, ‘Hashi6-20-1’ the five black grain wheat lines morphological features are common wheat, black grain, and black endosperm. Chromosome number of root tip was 42, and excellent genetic merit.The results of molecular markers shows that: the specific primers of E-genome is used to amplify the specific fragment of E-genome with size 982 bp from black grain wheat of ‘Hashi3-37-1’, ‘Hashi3-39-1’, ‘Hashi3-13-1’, ‘Hashi10-38-1’, ‘Hashi6-20-1’. Amplifying the five lines by specific SSR primer of E-genome, in these five lines, Thinopyrum elongatum 5ES chromosome ingredient approximately with the size 244 bp were amplified by primers Xedm 205, and the primer Xedm 335 amplifys fragments Thinopyrum elongatum 5EL chromosome ingredient of approximately 152 bp. It is preliminarily concluded that these five lines contain the genetic constitution of 5E chromosome genetic component. GISH shows that: there are E-genome chromosomes with the hybirdization signals in this five lines. In addition, cloning of F3 H which participates anthocyanin biosynthetic pathway, bioinformatics and gene expression analysis results show that: F3 H genes are highly conserved in black grain wheat and bread wheat, there is a small difference between nucleotide sequence and amino acid sequence, while that is obvious in expression patterns.