| Bacillus amyloliquefaciens is a kind of rhizosphere-promoting bacteria,which has the functions of promoting plant growth,enhancing plant stress and resistancing disease.Long non-coding RNA is a kind of RNA transcript with many biological functions and is a research hotspot in the field of plant biology.In this study,we used the Colombian ecotype Arabidopsis thaliana and Bacillus amyloliquefaciens 13-24 strain as experimental materials.The physiological and biochemical indexes of Arabidopsis thaliana roots under control,Bacillus amyloliquefaciens 13-24 strain treatment,high calcium treatment,high calcium + strain conditions were determined,which showed that the Bacillus amyloliquefaciens 13-24 whether enhance Arabidopsis thaliana resistance to high calcium stress.Whole transcriptome sequencing analysis was conducted to screen Arabidopsis thaliana root lnc RNA whose expression quantity were affected by high calcium stress and strain treatment,analysis of the biological functions of these lnc RNA;The mechanism of action of Bacillus amyloliquefaciens 13-24 to enhance the resistance of Arabidopsis thaliana to high calcium stress and the possible function of lnc RNA in this process were preliminarily revealed.The preliminary results of this study are as follows:1.This study measured Arabidopsis thaliana root length and shoot dry weight,root potassium and sodium ion content,oxidative stress related indicators and antioxidant enzyme activities under four culture conditions,namely control(Control),strain treatment(Bacillus amyloliquefaciens 13-24),high calcium treatment(40m M Ca Cl2),high calcium + strain treatment(40m M Ca Cl2+13-24).The results showed that 40 m M Ca Cl2 treatment significantly inhibited the growth of Arabidopsis thaliana roots,while Bacillus amyloliquefaciens 13-24 treatment alleviated the inhibition of Arabidopsis thaliana growth by high calcium stress;40m M Ca Cl2 treatment severely destroyed the potassium and sodium ion balance of Arabidopsis thaliana roots,with sodium ion content increased,potassium ion content decreased.Bacillus amyloliquefaciens 13-24 treatment can reduce sodium ion content,increase potassium ion content,increase root potassium to sodium ratio;40m M Ca Cl2 treatment caused very strong oxidative stress,Bacillus amyloliquefaciens 13-24 treatment can reduce the content of MDA and H2O2,enhance the activity of some antioxidant enzymes,and greatly alleviate the oxidative stress caused by high calcium stress.Based on the results of comprehensive growth and physiological and biochemical indicators,it was concluded that Bacillus amyloliquefaciens 13-24 can improve the resistance of Arabidopsis thaliana to high calcium stress.2.RNA transcriptome sequencing was performed on 12 root samples of Arabidopsis thaliana under four treatments.A total of 0.94 Gb of Clean reads and 141.2 Gb of Clean base were obtained.And an average of about 78 Mb Clean reads and 11.15 Gb Clean base were obtained for each sample.The average GC content is about 47.38%.The Clean reads of each sample was compared with the reference genome of Arabidopsis thaliana.The mapping rate was between 98.4% and 99%,and the average mapping rate was 98.68%.These results indicated that transcriptome sequencing was of good quality and the data can be used for subsequent analysis.3.With the Log2 FC threshold of 1 and FDR>0.05 as the screening standard for differential expression of lnc RNA,the differentially expressed lnc RNA of four treatments was analyzed.The total number of lnc RNAs differentially expressed by Ca Cl2 VS Control,13-24 VS Control and Ca Cl2+13-24 VS Ca Cl2 were respectively 362,40,66.The number of up-regulated lnc RNAs was 145,13 and 20,respectively,the number of down-regulated lnc RNAs was 217,27 and 46,respectively.4.The expression trends of differentially expressed lnc RNA transcripts under Control,Ca Cl2 and Ca Cl2+13-24 treatments were classified and analyzed,and 8 expression patterns were obtained.Among them,the fifth expression pattern has 115 lnc RNA,this expression pattern indicated that lnc RNA was up-regulated by high calcium stress,but with the high calcium + Bacillus amyloliquefaciens 13-24 treatment,its expression returned to the control level.This study speculates that these 115 lnc RNA may play an important role in the response of Arabidopsis thaliana to high calcium stress and Bacillus amyloliquefaciens to alleviate the damage of Arabidopsis thaliana by high calcium stress.5.Genomic position analysis,weighted gene co-expression network analysis and GO enrichment analysis were used to predict the function of Bacillus amyloliquefaciens regulation and high calcium response lnc RNA.The predicted results revealed some genes that may be regulated by lnc RNA,such as Gibberellin-regulated family protein,Jasmonate-zimdomain protein 3,and protein kinase superfamily protein,etc.These genes were involved in biological processes such as oxidation-reduction,hydrogen peroxide metabolism,signal transduction,plant hormone regulation,and protein phosphorylation.This study speculates that the regulation of Bacillus amyloliquefaciens and high calcium response lnc RNA may play a role in these pathways and play a regulatory role,and improving the resistance of Arabidopsis thaliana to high calcium stress,and then influencing the physiological phenotype changes of Arabidopsis thaliana under high calcium stress. |
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