| In this experiment some high-quality parents have good HMW-GS such as Glu-A12*, Glu-B1x14+y15, Glu-D1x5+y10 and parents with high yield and disease resistance were crossed with Taigu-male-sterile wheat to establish a base population. The composition of HMW-GS in recurrent selection population after mutually-crossing three times was analyzed using SDS-PAGE and PCR method. Some agronomic characters of C3 and C4 recurrent population were also analyzed. The major results were presented below:1 New types of HMW-GS combination different from the parents were created such as "2*,7+8,5+10"and"1,14+15,5+10"etc. Many kinds of HMW-GS polymers such as "14+15,17+18", "7+8,14+15", "2+12,5+10"etc. were found in the population. "2+10"and "5+12"were the HMW-GS recombination types because the linkage of "5+10", "2+12"had been broken. But in the population the "null,14+15,2+12" HMW-GS combination had the highest frequency(13.2%), and in Glu-D1 loci the subunit type mostly were "2+12". This showed using Taigu-male-sterile wheat recurrent selection is an effective way to create new subunits types, olao higher selection pressure should be given to the population because the subunit combination mode were still not reasonable. 2 The HMW-GS of the fertile and the sterile in recurrent population showed different rules. The fertile is 1.9 times purer than the sterile in HMW-GS. The seeds of the fertile were mainly the pure type, while the sterile were mainly the mixed type. As for as the seed of an individual plant was concerned, the HMW-GS of the fertile seed was still not pure and showed high otherness, although the seeds were come from self-crossing. This is in accordance with the genetic inherited rules of other agronomic traits. 3 Preliminarily formed a fertile population cherish high-quality HMW-GS. By half-seed-method 80 seeds have good HMW-GS compositions were selected from 297 seeds of the fertile plant. We can select good cultivates and idioplasms from it. 195 seeds were approved have good subunits, the remainder of these seeds were planted and formed the C4 recurrent population. 4 The proper amplification system of primer1 and primer2 was found out. Amplification system: 2.5μL 10×Buffer, 1.5μL MgCl2 (2.5mM), 2μL dNTP(2.5mM),2×4μL primer(2μM, about100ng),4μL DNA(about 50ng),02μL TaqE(5U),6.8μL ddH2O, total volume:25μL As a specific molecular marker of 1Dx5 gene, the primer showed strong specificity. The exceptionally different belts of 450bp could be amplified steady. 264 individual plant were proved have high-quality subunits 1Dx5 from 433 samples, with frequency of 60.97%. These two primers can be used in molecular marker assisted selection. The proper amplification system and cycle parameters of the specific primer of Glu-D1y10(12) and Glu-B1 14+15 gene has not found. So primer3 and primer4, primer5 and primer6 can't be used in wheat breeding practice for the moment.5 The average plant height of recurrent population was below 75cm, and the fertile' height was higher than the sterile'. The plant height difference had high correlation with inter-cross seed set rates of the sterile. The difference of C3 and C4 recurrent population was 3.51cm, 1.61cm respectively, their inter-cross seed set rates was 55.66% and 17.85% correspondingly. |
