The Marker Screening Of Temperature Sensitive Gene Of The Male Sterile Gene In Thermo-Sensitive Male Sterile Line Of B.Napus Of 104S

Male sterility (CMS) is one of the main way in rape heterosis utilization. ecological male sterile male sterility (CMS) is a new important heterosis utilization way in recent years.It has been more and more applied in rape hybrids use. ecological male sterility has a wide range of restorer, easy screening strong advantage combination and can be achieved in the breeding work "a series of dual-use" simplify the process of breeding and reduce the cost of production.Currently, rape male sterility in breeding ecology already has initial success. Two-line hybrid Brassica napus varieties such as Xiangzayou-5 and Ganliang you-2 et al has application in rapeseed production.Ecological male sterility has good prospects for development in rapeseed production.But ecological male sterile Brassica started late and research has lagged behind and sterile mechanism on ecological male sterile Brassica unclear, especially targeting eco-sterile genes and gene cloning and functional studies still in its infancy.Therefore, not only restricts the use of the value of ecological male sterile CMS line rapeseed but also limits the development of two-line hybrid rapeseed.Thermo-sensitive genie male sterile (TGMS) line104S was originally discovered in the paternal 99104 cut stump regeneration in a fertility variable plant, most of the flowers had been completely sterile flowers when found and only a small number of flowers were fertile flowers included semi-sterile flowers, these self-fertile flowers artificial pollination and obtained a small amount of seeds.A year after selected plant bagging selfing and fertility transformation period to 2004 and agronomic traits were stable and consistent group named 104S.A completely sterile plants had been found in fertility variable group.And hybridize with the temperature-sensitive plant.the seeds were planted into lines.A year after used the temperature-sensitive plant and sterile plant hybridization in pairs.Obtained temperature-sensitive plant and infertile plant basic ratio of 1:1 group named 104SA.104SA is separated from the 104S.So 104S and 104SA can be seen as a pair of near-isogenic lines.To 2015 the near-isogenic line had 12 generations, near-isogenic line had been purified.In this study, to make further analysis of themechanismof inheritance and fertility transition of 104S, artificial climate chamber test and screening of SSR molecular markers were conducted.Meanwhile, it could evaluate the value of male sterile line by investigated the temperature reaction of 104S.and make foundation of the fine location even cloning of the male sterility gene by means of screening molecular markers to find the linkage group of male sterility gene of Huiyou50S.The main results are as follows:1.Through investigating 104SA female parent group sterility.Parental groups has a total of 180 plant,of which there are 76 sterile plants and104 temperature-sensitive plants.Sterile strain:Temperature-sensitive strain ratio was 0.73:1.The normal infertility strain and temperature-sensitive strain ratio should be 1:1.So, subsequent near-isogenic lines of breeding work 104SA need to continue.2.In artificial climate chambers test,five different temperature conditions were set.The results showed that,In the 12? and14? treatment,the flowers of 104S were fertile.In the 22? treatment,the flowers were sterile.In the 16? treatment,most of the flowers were fertile,stamens were missing of only one petal.In the 18?treatment,the Flowers were not only sterile but also fertile.Also there were some stamens with trace pollen in one flower.In the 20? treatment,most of the flowers were sterile,and a small number of fertile flowers which stamens smaller and only a trace of pollen.It showed that the critical temperature of the sterile line was not a fixed value,but a range.It detected that the temperature-sensitive of themale sterile line period was 12 days before flowering.3.In the screening of 799 paris of SSR molecular markers,the male 104S,the female 104SA and F1 were used for research materials.There were two pairs of polymorphic primers,CNU398 and BRAS095,and two pairs of SSR primers were above in 5 chain group.Then according to the 5 chain group next to two pairs of polymorphic primer and intermediate above 13 primers for the male and female parents and F1 for validation,and selected three pairs of polymorphic SSR primers,and the three SSR markers were between CNU398 and BRAS095 which had short distance withBRAS095.