Protoplast Of Triticum Aestivum L And Aegilops And The Effect Of Exogenous Material On Triticum Aestivum L

Wheat (Triticum aestivu) is one of the world's most important food crops. New varieties are being developed to adapt this grain to various abiotic and biotic stresses. Genetic resistance is the most effective, economical, and environment-friendly method of managing stresses. One way of increasing the genetic variability of wheat is by somatic hybridization, which can help to introduce new characters from other species or genera.In this context, the Aegilops is a wild relative of wheat that possesses several interesting agronomic traits of disease resistance. As these species are cross-incompatible, the only way to obtain such intergeneric hybrids is protoplast fusion and somatic hybridization. In this study, embryo of Triticum aestivu and leaf of Aegilops were taken as experimental material. The protoplast was stained with neutral red(NR), FDA and Rhodamine B and tested fusion rate during different density(1×106,5×106,1×107,5×107per mL). The best protoplast fusogen and washing solution condition were selected via an orthogonal experimental design. The result showed that:Embryo of wheat was stained with FDA, and leaf of Aegilops was stained with Rhodamine B. The best time of enzymatic hydrolysis was 4 hours and the best of density was 1×107 per mL. PEG6000 content(w/v) 25%,0.50 M sucrose, pH 5.8 and 5.5 mM CaCl2 were the greatest of protoplast fusogen and washing solution condition.5-azacytidine was DNA demethylating agent and has been used in the studies of plant epigenetics that based on DNA demethylation. Some usefully epigenetic traits that we related to undermethylation were taking Potential breeding value. CPPU is a very important kind of synthetic cytokinin belonging to plant growth regulators. It can promote cell division and cell enlargement, delays plant senescence, etc. At present CPPU was widely used on fruit and melon vegetable, but seldom on wheat. In order to investigate the effects of 5-azaC and CPPU on wheat, DNA methylation was tested by HPLC. This study can provide the basis for realizing the effect of 5-azaC on the seedling morphology and growth and development of wheat and laid a foundation for exploring the effects of foliar application of 5-azaC and CPPU on gene expression in different periods, and it had an important significance theoretical and practical for improving the wheat yield. The result was showed that:After treatment with low concentration (0,5,10,50,100μM) of 5-azaC, the methylation of the wheat(paksaylabgg, Taichung29 and Taichung29*6/CarstensV)DNA was reduced.5-azaC was sufficient to induce flowering.1000μM of 5-azaC caused demethylation and made seedling death in 20 d.Foliar application of 5-azaC can delay leaf senescence, reduced the methylation of DNA in flag leaf. Application of 5-azaC soon after anthesis can improve the remobilization of stem reserves, and slightly increase the content of protein and gluten of the seed.Foliar application of CPPU can delay leaf senescence, reduced the methylation of DNA in flag leaf. Application of 5-azaC soon after anthesis can increase per steam grain weight, promote water soluble carbohydrates transport to sink, and slightly increase the content of protein and gluten of the seed.