Establishing Of Highly Efficient Regeneration System And Cloning Cellulose Synthesis Related Gene Of Triarrhena Lutarioriparia L.Liu,Sp.Nov.

Triarrhena lutarioriparia L.Liu,sp.nov. is a specific new species in Triarrhena of China. Being useful in soil and water conservation, dyke strengthening and flood control, water and air purification as well as natural eco-system protection.Triarrhena lutarioriparia is C4 plant, which is a cellulose-ethanol raw stuff as well as sustainable high biomass resource. As a result, cultivating and exploiting Triarrhena lutarioriparia in large area are very important. Regulating gene expression of cellulose synthesis in Triarrhena lutarioriparia by genetic engineering can hopefully increase the content of cellulose, improve its quality and produce new germplasm resources.In this paper, young ears of Triarrhena lutarioriparia were used as materials. The high frequency regeneration system and genetic transformation of Triarrhena lutarioriparia were studied. Mean while, cellulose synthesis related genes CesA and UGAD were cloned. And then analyzed by bioinformatical methods. The main research results are as follows:1. Establishment of high frequency regeneration system of Triarrhena lutarioriparia. In the dark conditions, the optimal induction medium for callus in this paper was MS medium with 2.0 mg·L-12,4-D,1.0 mg·L-16-BA,0.5 g·L-1 Casein acids hydrolysate and 0.5 g·L-1 proline. The optimal differentiation medium for bud was MS medium with 0.5mg·L-1 NAA,0.5 mg·L-1 KT,0.5 mg·L-1IAA,2.0 mg·L-16-BA.0.5 g·L-1 Casein acids hydrolysate and 0.5 g·L-1 proline. The optimal rooting culture medium was MS medium with 1.0 mg·L-1 NAA and 0.25 mg·L-1Met. The induction rate of young ears of Triarrhena lutarioriparia was 100%, the differentiation rate was 92.72%, the rooting rate was 92.42%.2. The transplant survival rate was improved. After acclimatization, the culture-bottle seedlings were transplanted into a growth mixture which contains nutritious soil and perlite(1:1). The survival rate was up to 100%.3. Exploration of genetic transformation system from Triarrhena lutarioriparia. Transgenic plants with GFP gene have been obtained. The integration of these genes in the transformed plants were confirmed by PCR.4. Through the molecular biology and bioinformatics technology, a part of CesA and UGAD gene sequences were cloned in Triarrhena lutarioriparia, the sequences were bioinformatically analyzed.