Study On Plant Tissue Culture Technology In Cultivating Rapidly And Directive New Strains Of Transgenic Wheat

Wheat is a major food crop for residents in China. Conventional breeding methods of wheat had long cycle and low efficiency. Therefore,it is becoming an objective requirement of new wheat species to short the breeding cycle and increase its speed.The research has used materials that wheat germplasm with TaEBP/DREB were donor parent and 10 main winter wheat strains in Huanghuai zone were acceptor parent,We used generation-adding under the condition of greenhouse,to cultivate rapidly and directive new strains and germplasm of transgenic wheat by technology that immature embryo directly culture,anther and microspore culture ,tracking and detection of molecular ;At the same time, the factors affcting transgenic wheat immature embryo culture,anther and microspore culture were investigated such as mediums,coagulants,time of pretreatment with lower temperature,kinds and concentrations of anxins and genotypes,The system of tissue cultureand regeneration were optimized. the main results are as follows:1. The research has established preliminary a breeding system that combine immature embryo directly culture,Green plant rapidly vernalization,greenhouse generation-adding with tracking and detection of molecular to cultivate rapidly and directive new wheat strains,and the rate of immature embryo germination was influenced by components of medium,kinds and concentrations of anxins,The result showed that MO medium with 0.5mg/L NAA and 0.1mg/L 6-BA is the best one.2. The system of wheat anther callus and plantlet regeneration were optimized. Callus induction rate of wheat anthers was influenced by components of medium,coagulant,time of pretreatment with lower temperature,kinds and concentrations of anxins.During the process of callus induction of anthers,callus induction rate of wheat anthers could be improved by following factors:IMI medium;CHB medium with agarose coagulant;the inflorescence treated 4d with 4℃lower temperature before culturing,and emerged a phenomenon that anthers regenerated directiy plantlet;CHB medium plus anxins 1.0mg/L(2,4-D),0.5 mg/L (KT) and 0.1mg/L(MET);Genotype is one of the key factors that had effect on anther culture, the rate of callus induction of 18 genotypes between 0.45% and 56.41%;Redifferentiation rate of wheat anther callus on IMR medium achieved at 63.64%. 3. The research of microspore culture indicated that: Microspore after isolation and purification brought about globular embryoids when they were cultured 18 days,Pear embryoids formed after culturing 24 days,and 1mm embryoids were produced after culturing 35 days; The average number of embryos of inflorescences which were pretreated in low temperature for 4d produced respectively more than which were pretreated in low temperature for 7d;At the same time,the same matetial produced more embryoids on IMI liquid medium than semi-solid one...