Construction Of Rhizobia Strains Which Were Labeled By Red,Yellow And Green Fluorescent Protein

In order to research the competition and growth-decline relations between the target and in digenous rhizobia,ascertaining the nodule occupancy and nodulationrate oftarge trhizobia,tracer technique is needed.Based on classic altri-parentalmating taking filter membrane,marked many rhizobia that contain the all of genetically stable RFP(Red fluorescentprotein)genes were obtained rhizobia.The research must be a differentcolor fluorescent protein RFP,YFP and GFP plasmid,first need to be successful import auxiliary bacterium escherichia coli,method based on the stability of the building and validation.The experiments based on nine kinds of fluorescence labeling bacteria were inoculated alfalfa gannong no.5(Medicagosativa CV.Gannong No.5)and alfalfa s.343(Medicagosativa CV.S.343),measured plant biomass,root length,plant height,leaf number,leaf area,individual nodulationnumber,nitrogenaseactivity,nodule occupancy and leafchlorophy content of inoculatedalfalfaplants.There sults were as follows:1.Many research results show that heat shock protein transduction plasmid can be used for transduction,and with the combination of e.coli.In this experiment,in the traditional heat shock transduction process,three kinds of color can stable and e.coli,shows that heat shock transduction for red,yellow,green fluorescent protein and the transfer of e.coli.2.tri-parental mating parent has played a very important role in carrier of genetic transfer material[11],different receptor bacteria may also affect the joint efficiency 2 of tri-parental mating.In this experiment,GFP fluorescence joint strain transfer rate are the highest,and the other colors differ significantly(P < 0.05),fluoresce-nce plasmid presented.according to also lower,presumed to green fluorescent tracer of plasmid in alfalfa rhizobia with the choice of the optimal conditions.3.Two varieties of alfalfa seedlings' biomass,nitrogenaseactivity,rootlength,planth eight and leaf chlorophy llconten to alfalfa plantsino culated with rhizobium inoc-u lation 3 kinds of color fluorescent tags and their fluorescence marked strainshad si gnificant difference with the treatment of noinoculation(P < 0.05),comparing the fl uorescence marked rhizobia with their corresponding original rhizobia,they did no harm to alfalfa plantand could expressstably inplant,and had the similar growth pr omotion ability with origin alrhizobia.