Screening And Analysis Of Drought Resistance Genes In Wild Cerasus Humilis Based On Stress Genomics

Wild cerasus humilis is a wild characteristic fruit tree growing in Inner Mongolia.It not only has rich germplasm resources,high nutritional value,but also has strong stress resistance,especially in drought resistance.The wild cerasus humilis originated from the arid region of Inner Mongolia Plateau.The long-term drought and harsh geographical environment made it have unique plant morphology and drought-resistant ability.At present,a large part of the research on drought resistance of cerasus humilis remains at the level of physiological and biochemical studies,and there are few molecular biological studies on the mechanism of drought resistance of cerasus humilis.Wild cerasus humilis cDNA library was constructed.High-throughput screening of drought tolerance genes in yeast;The cloning and bioinformatics analysis of drought stress related genes of cerasus humilis were studied at the molecular level of drought resistance.This study provided important guidance for the screening of drought-resistant varieties of cerasus humilis,and also provided an important basis for further exploration of its molecular mechanism.The final important results were as follows:(1)Through drought stress,a uniform cDNA library of Prunus sativa was constructed for the study of wild Prunus sativa leaf tissue.The obtained storage capacity is 4.52×10~7cfu/m L,with a total clone count of 9.04×10~7 cfu.We conducted PCR identification of the colonies using random sampling of 24 clones on a plate.The average size of the library fragment was 1000 bp,and the recombination rate was 96%.(2)Using yeast high-throughput screening to screen drought resistant genes,96 positive yeast clones were selected,all of which were amplified by PCR.After Seqman and BLAST alignment,empty and duplicate sequences were removed,and 44 different sequences were obtained.Through mannitol simulation of drought resistance,the 44 cloned genes were subjected to rotational validation.All 44 clones were able to grow on SG-U,SG-U+100 m M mannitol(1:10000)plates.(3)After comparative analysis,we found that 44 genes and proteins may be responsive to drought stress in Cerasus humilis leaves,and these responses may be significant.This article amplified the sequences of ChU40rps27 a and ChTp258 genomes with drought resistance.The open reading frames of ChU40rps27 a and ChTp258 were 471 bp and 231 bp,respectively,encoding 157 and 77 amino acids.(4)Nucleotide sequence consistency and evolutionary tree analysis showed that the highest consistency between ChU40rps27 a and Apple 40S(XM＿050288755)was 88.5%.The evolutionary tree results show that ChU40rps27 a has the closest evolutionary relationship with citrus(XM＿006487968).The highest nucleotide sequence consistency between ChTp258 and peach(XM＿020567676)was 94.4%.The evolutionary tree results show that the evolutionary relationship between ChTp258 and peach trees(XM＿020567676),apricot trees(XM＿034370534),plum trees(XM＿008244122),and sweet cherries(XM＿021951344)has recently clustered in the same branch.