Cloning And Function Of ERF106a In Response To Stress In Cauliflower

Brassica oleracea L.var.Botrytis belongs to the Brassica oleracea L.variety,an important vegetable and horticultural crop.It has a fresh taste and is rich in vitamins,minerals and sulforaphane and other anti-cancer and anti-cancer active ingredients.Loved by consumers.As an exotic breed in China,cauliflower has a shortage of germplasm resources,,a narrow range of suitable temperature for reproduction,a poor adaptability to adverse environment and a vulnerability to adversity.Therefore,revealing the molecular mechanism of maladaptive cauliflower is of great value for breeding new varieties of cauliflower with high stress tolerance.Based on this,the purpose of this study is to separate,clone,verify the function of ERF transcription factors in cauliflower,and preliminarily explore the mechanism of adversity response,so as to provide theoretical guidance for further revealing the function of ERF transcription factors in cauliflower and for anti-broccoli molecular breeding.The main research results are as follows:1.The full-length c DNA of ERF106 a,ERF056,ERF003 a and ERF036 genes of cauliflower were isolated and cloned,and amino acid analysis showed that they all belonged to AP2/ERF superfamily.2.Analysis of transcriptional expression characteristics confirmed that: cauliflower ERF106 a had the highest expression level in cauliflower,and the expression level in different tissues increased with development time;the expression level of ERF106 a in 4 week-old cauliflower seedlings was up-regulated under different stress treatments,and its promoter region contained A variety of abiotic and biological stress related cis-elements suggest that it may respond to a variety of abiotic stresses.3.Successfully constructed ERF106 a overexpression vector for caulifloweri.Nine ERF106 a overexpressing cauliflower transformant lines and homozygous lines overexpressing ERF106 a transgenic Arabidopsis were obtained.4.Analysis of transgenic and wild-type overexpressed ERF106 a showed that the transgenic Arabidopsis thaliana had early germination and strong germination;the angle of the rosette leaf position was small,and the leaf type changed;bolting time was about 7 days late,and there were many branches,small inflorescences,many The flowers are large;the root length and the number of lateral roots are large,showing a stronger ethylene response.Transgenic cauliflower grows fast,with large leaves and well-developed roots.5.Adversity stress analysis showed that ERF106 a overexpressed transgenic Arabidopsis sprouts early,with long roots and many lateral roots.Tissue staining showed that transfected Arabidopsis thaliana was lightly stained after staining with NBT,DAB,and Evans Blue.Natural drought and salt stress indicate that the survival rate of transgenic Arabidopsis plants is high.All of the above reveals that ERF106 a overexpression transgenic Arabidopsis has strong resistance.The stress of cauliflower tissue culture seedlings showed that ERF106 a overexpressed cauliflower grew well and had strong resistance to adversity.In vitro leaf experiments showed that the overexpression of ERF106 a cauliflower leaves had slow water loss,low stomatal density and stomatal conductance;under Na Cl and MV treatment,the leaves showed less chlorosis and less disease after black rot inoculation,indicating that the transgenic cauliflower had high resistance.6.Sequencing results of cauliflower transcriptome showed that a total of 9460 differential genes were screened,of which 4433 were up-regulated and 5027 were down-regulated.The results of GO and KEGG enrichment are mostly stress-related genes or pathways,suggesting that cauliflower ERF106 a is involved in regulating the expression of stress-related genes.