Jujube Germplasm Resource Collection,Preservation And Genetic Transformation Of ZjTCP6 And ZjTCP16 Genes

Ziziphus jujuba,a plant of Rhamnaceae,has a high nutritional value and is the first fruit tree that can be used as both dried and fresh fruits.The study of Chinese jujube has a certain history,mainly in the survey,description,collection,evaluation and other aspects of the germplasm resources of jujube.The establishment of germplasm resource management platform can lay a foundation for the rapid development of jujube planting industry.This article through to the henan region jujube germplasm resources collection,adopt the method of grafting jujube germplasm nursery is established,using the germplasm resource management system platform,according to the jujube germplasm resource description specification and data standard "nursery of germplasm resources of resources investigation,survey data entry system to record,provide favorable platform for jujube resource sharing.Tissue culture technique was used to preserve'honey pot' and 'jujube jujube' in vitro.Jujube disease is a disease often occurred in the cultivation of jujube trees.It is also a devastating disease caused by phytoplasma infection.Phytoplasma is a plant pathogen lacking cell wall,which is parasitic in the phloem sieve of infected plants.TCP(TEOSINTE BRANCHED 1/CYCLOIDEA/PCF)transcription factors are specific regulators of plant development and morphogenesis,and studies have shown that TCP transcription factors are protective against phytoplasma infection.By means of genetic transformation,gene function was verified by over-expression in jujube disease seedlings,which provided theoretical basis for the research on occurrence mechanism and prevention of jujube disease.The main results are as follows:(1)through the field investigation of puyang city,yongcheng shuangqiao town,lingbao city,xinzheng jujube research institute,more than 100 germplasm resources of 40 varieties were collected,and the germplasm resource nursery of jujube was established by grafting.(2)the in vitro preservation system of the collected'honey pot' and 'jujube' was constructed,and the effects of different plant growth regulator combinations on germination rate,proliferation coefficient and average seedling height were compared.The results showed that the germination rate of 'honey pot' jujube with embryo as explant was 91.2%when GA3 was 1.0 mg/L,and the germination rate was 92.5%when stem segments of 'jujube puree' were used as explants.The growth regulator combination was MS+KT 2.0 mg/L+NAA 0.3 mg/L.The maximum proliferation coefficient was obtained by MS+6-ba2.0mg/L+iaa0.1mg/L treatment.In the average seedling high school,the average seedling height was the highest in the combination of 6-ba1.0mg/L+ iaa0.5mg/L.(3)a date germplasm resource information management platform was established.The database was composed of 7 modules,including 96 fields.Nursery for germplasm resources preservation of fresh food,dried,view and admire,combination USES four types of jujube varieties of different traits investigation,investigation including neak traits,infarction depression depth,cross the fruit,fruit size,leaf shape,leaf 23 properties such as gloss,utilization of germplasm resources and crop management system platform phenotypic analysis system of survey data entry system,survey data received 700 groups of characters,122 groups of character images recorded,to gradually establish a perfect jujube germplasm resources information platform to lay the foundation.(4)the regeneration system of jujube disease seedlings was established.The effects of different explant sources,different plant growth regulator combinations and concentrations on germination rate,proliferation coefficient,average seedling height and rooting of jujube disease seedlings were compared.The results showed that the survival rate of hydroponic bud of dormant branches was higher than that of diseased stem in field.The growth regulator with the highest germination rate was MS+kt2.0mg/L+ naa0.1mg/L,the optimal combination of proliferation medium was MS+6-ba2.0mg/L+ naa0.3mg/L,the medium of strong seedling was MS+kt2.0mg/L+naa0.3mg/L,and the medium of rooting was 1/2MS+naa0.5mg/L.PCR detection of phytoplasma in tissue cultured seedlings after 3 to 5 generations showed that phytoplasma was stable and parasitic in the regenerated seedlings.The established tissue culture system of jujube blight seedlings can preserve the phytoplasm of jujube blight in vitro.(5)the genetic transformation of ZjTCP6 and ZjTCP16 were carried out.TCP transcription factor family according to the early in the disease process in the expression of process analysis,screening ZjTCP6 associated with disease occurrence and ZjTCP16 two transcription factors,and the date of the plant of chlamydial infection somaclone of agrobacterium mediated genetic transformation,through infection incidence of plant stem section,trained to deal with different time,found that different co-culture time has a certain influence on genetic transformation efficiency of jujube,among them,at the time of co-culture 3 d conversion efficiency is highest,based on the screening of cultured from the PCR identification of seedlings,PCR positive plants of ZjTCP6 and ZjTCP16 were obtained.The results of this paper provide reference for the protection and utilization of jujube germplasm resources,the occurrence mechanism and prevention of jujube disease.