Study On The Editing Sites In The Transcript Of Mitochondrial Atp6 Gene Of Wheat With Aegilops Kotschyi Cytoplasm

Male sterility of wheat with Aegilops kotschyi cytoplasm refers to those sterile lines which contain the cytoplasm of Aegilops kotschyi. Prominent traits of these lines are that they can be easily kept sterile completely, restored and the restore resources are widely distributed in many wheats.This CMS is a good material of research male sterility and heterosis utilization. It also has a good potential and prospects of practical application. Using cytoplasmic male-sterile line ms(Kots)-90-110 (A) and the Near-isogenic Line BC5F2 which are fertile as the plant materials,we identified the genetic consistency between NIL and CMS,according to the difference of gliadin.ensuring the genetic consistency are highly,we analyzed the editing sites and the friquences in the conservative area of mitochondrial atp6 gene transcripts to provide the theoretical basis of male sterility and heterosis utility.The main results obtained are as follows:1. With polyacrylamide gel electrophoresis (A-PAGE) methods, we analyzed the wheat gliadin of 17 samples, which containing cytoplasmic male-sterile lines, near-isogenic lines, parents and the control of Chinese Spring. There were 24 the relative electrophoretic mobility of different bands. Each material coud separate 16 to 18 bands,of which nine bands (50%) were common and nine were polymorphic to all tested materials.The similarity coefficient were 0.967 between the cytoplasmic male-sterile line and NIL.Can be seen through the 5 generations of backcrossing and selfing generation, the near-isogenic lines with high genetic similarity of male sterile lines for follow-up study of RNA editing in addition to fertility, the nuclear transfer are quite consistent with the background research material.2. RNA editing of atp6 gene in different stage of pollen development was analyzed by direct and clone sequencing using cytoplasmic male-sterile line ms(Kots)-90-110 (A) and the Near-isogenic Line BC5F2 as the experimental materials.The results indicated that DNA sequences of cytoplasmic male-sterile line ms(Kots)-90-110 (A) and the Near-isogenic Line were completely unanimous,and the homology are 100% comparing with that of Triticum aestivum and Triticum timopheevi. 3. Compared DNA sequences to cDNA sequences, the conservative area of atp6 gene transcripts were found that there were 15 editing sites between male sterile line and fertile line,respectively,and all the sites were completely unanimous by clone and direct sequencing. Fifteen editing sites occurred at the 1st or the 2nd position of codons ,which may alter the amino acid type, two occurred at the 3th position of codons which can't change the amino acid type and the 6th and 7th sites were co-transcript. The frequency of editing sites were gradually increasing with the development of anther stage.4. The restorer gene in BC5F2 obviously increased the editing frequency of each editing site by comparing the sequence of atp6 gene between male sterile line and fertile line.Transcript inadequately edited may affect the normal mitochondria function.It is concluded that RNA editing of atp6 might have a certain relationship with wheat CMS with Aegilops kotschyi Cytoplasm.