Black Chokeberry And Blueberry Isolation And The Protoplast Fusion Research

Black chokeberry(Aronia melanocarpa) is a new small berry fruit.As Black chokeberry to be of value in dominant gene restrictions,making improvements through conventional breeding methods cultivars be greatly restricted.Black chokeberry rich of fruit anthocyanins,cold hardiness,pest resistance capability.Blueberry fruits of a growing emphasis on health function,in addition to prevent brain aging,enhance heart function, anti-inflammatory anti-cancer,anti-oxidation also can be effective and have good eyesight protection.Use of technical means,blueberry and black fruit gland rib Sorbus the protoplast fusion,it is possible to overcome the natural world in the form of reproductive isolation between species,thereby creating a new fine of resources.This test use black chokeberry and midbush Blueberry 'Northland' leaf as material,the study of the enzyme isolated protoplasts of composition,hydrolysis time and method of solution pH,osmotic Pressure regulator concentration,centrifugal conditions,leaf pretreatment methods,CPW solution components and in vitro enzymatic hydrolysis former medium of change on protoplast Purification of the best conditions in these two plants and materials for a preliminary protoplast the integration of research,and adeno-ribbed fruit and blueberry Sorbus somatic cell hybridization preliminary exploration.Test main findings are as follows:1 The black chokeberry protoplasts isolated conditions:25℃±1℃,cellulase,pectinase,mannitol 0.7 mol / L +5 mmol / LMES,hydrolysis time 8-10 hours a pH value of 6.5-7.2 Cut the black chokeberry top 1-4 leaf into 1-2 mm long,enzymatic hydrolysis better than under the skin torn off leaves good.Hydrolysis the same time,the use of static and the role of dark hydrolysis 30 r / min dark double enzyme oscillation there was no significant difference,you can use a simple static Enzymatic. There will be no vaccine hydrolysis before dark handle it reduces the output of protoplasts and vitality.3 Osmotic strength of the output restrictions protoplast role was clear,0.7 mol / L mannitol under black chokeberry protoplast yield very different and adjacent processing.4 The black chokeberry hydrolysate of nearly neutral pH value,the enzyme effective.In nearly neutral pH range,after enzyme hydrolysis of pH values similar rate of decline.5 CPW formula added Mg²⁺ can effectively improve the living Blueberry protoplast yield.6 With the increase in centrifugal time,Blueberry protoplast yield higher,but the protoplasts increased activity decreased after first.7 Blueberry protoplasts isolated in the operation of adding pectinase,protoplast yield improved to 0.5 g / L pectinase by protoplast live up to 3.34×10⁵ / g FW8 Blueberry vitro enzymatic hydrolysis of sucrose content in the training before 20 g / L medium,the largest live protoplast yield to 4.61×10⁵ / g FW.9 Observed that a single protoplast adhesion process,protoplast fusion linked to the membrane,has not formed a complete nuclear differences of,but not conducive to identify low-temperature protoplast fusion.