Study On The Molecular Mechanism Of Prickly Development Of Zanthoxylum Armatum

Zanthoxylum armatum is a plant of Zanthoxylum in Rutaceae.It has the advantages of rapid growth,strong ability of soil and water conservation,early bearing and high yield.Its fruit can be used as food seasoning and traditional Chinese medicine,which has good economic value and ecological benefits.However,the surface of Zanthoxylum armatum is densely covered with prickles,which causes some troubles to the planting,management,harvesting,processing and other processes of Zanthoxylum armatum,and there is no good solution at present.This study attempts to explain part of the molecμLar mechanism of prickly development of Zanthoxylum armatum from the perspective of morphological observation and genetics of molecμLar biology,which provides a theoretical basis for the future development of Zanthoxylum armatum with little or no prickles by genetic engineering.Firstly,the branch diameter,number of prickles,height of prickles,length and width of prickles of 120 branches of 30 plants of 5 grafted strains and 1 seedling strain were measured and analyzed statistically.Secondly,we analyzed the transcriptome data of the leaves and stems on the young shoots of the grafted and seedling trees of Zanthoxylum armatum,screened out the CCR and CHS genes that may be related to the prickly development of Zanthoxylum armatum,and then designed specific primers according to the selected gene sequence to clone the genes.Then,NCBI,DNAMAN and other software were used for bioinformatics analysis of the cloned CCR and CHS genes.Finally,the specific expression of CCR and CHS genes in the organs of grafted and seedling trees was detected by fluorescence quantitative PCR.The resμLts are as follows:1.The statistical data of 5 characters showed that the variation coefficient of prickle height is the largest,followed by prickle number,prickle length and prickle width,and the variation coefficient of branch diameter is the smallest.2.Analysis of variance and mμLtiple comparisons showed that there are significant differences in the length,width,height and number of prickles after grafting,except for branch diameter.No matter in the length,width,height or number of prickles,there are significant differences between the control group(seedling trees)and the experimental groups(grafted trees),but most of the differences between the experimental groups are not significant.3.Correlation analysis showed that prickle length,prickle width,prickle height and prickle number are significantly positively correlated with each other,and they are not correlated with branch diameter.4.Through the analysis of transcriptome data,a CCR gene and a CHS Gene which may be related to the prickly development of Zanthoxylum armatum were screened out.The full-length c DNA sequences of CCR and CHS genes of Zanthoxylum armatum were successfully cloned,named Za CCR and Za CHS,with NCBI ID of MK953726 and MK953733,size of 990 bp and 1173 bp respectively,encoding 329 and 390 amino acids.5.The relative molecular weights of Za CCR and Za CHS are 35680.65 Da and42710.46 Da,respectively,and the theoretical isoelectric points are 5.93 and 6.35,respectively.The results of protein sequence analysis showed that Za CCR contains four CCR functional domains,Za CHS contains two highly conserved CHS sequences and two CHS functional domains.6.The CCR similarities between Za CCR and Citrus sinensis,Citrus unshiu,Citrus clementina and PopμLus tomentosa are 86.32%,86.02%,85.71%,71.00%;the CHS similarities between Za CHS and Citrus sinensis,Citrus clementina,Citrus unshiu and Citrus suavissima are 80.51%,80.25%,80.00%,79.49%.Phylogenetic tree analysis showed that Za CCR is closely related to the CCR of Citrus sinensis,Citrus clementina and Citrus unshiu,and Za CHS is closely related to the CHS of Citrus sinensis,Citrus clementina and Citrus unshiu.7.The results of fluorescence quantitative PCR showed that the expression of Za CCR from high to low are: grafted stem,seedling stem,grafted leaf,seedling leaf;the expression of Za CHS from high to low are: grafted leaf,grafted stem,seedling stem,seedling leaf.