Mulberry Heavy Metal Zinc Stress Transcriptome?Cloning Of Related Genes And Expression Analysis Of Differential Genes

Zinc(Zn²⁺)is an essential nutrient for mulberry,It plays a key role in biological growth,development and pest and diseases prevention.It is based on zinc participation of enzyme catalysis,protein synthesis,carbohydrate and auxin metabolism,energy dissipation,environmental stress tolerance,etc.In addition,it plays a variety of roles in the biochemical process.It is the catalytic and structural protein cofactor of hundreds of enzymes.It has key structural functions in protein domains of interacting with other molecules.Excess or inadequate zinc will cause different degrees of damage to mulberry.This study uses annual Fengchi mulberry seedlings as materials to explore the cloning and analyze the heavy metal zinc stress-related genes from the field of molecular biology.Moreover,the study employs molecular biological techniques,such as RT-PCR and genome sequencing,to obtain two full-length flavonoid c DNAs Sequence and analyze the credit information.After the treatment with different concentrations of Zn²⁺stress conditions,the detection of physiological and biochemical related indicators and second-generation gene transcriptome sequencing are conducted to understand the zinc metabolism of mulberry from the molecular mechanism.Therefore,the thesis is expected to further elucidate the molecular mechanism of high-efficiency absorption and utilization of zinc ions by plants.Moreover,the cultivation of zinc efficient use of new varieties provides an important basis in the field of molecular biology,aiming to improve the utilization efficiency of mulberry zinc fertilizer and maintain the sustainable development of the ecological environment.The main research contents and results are as follows:1.Determination of physiological and biochemical indicators of zinc stress:Under the stress of different concentrations of zinc,the measured physiological indicators of mulberry trees are:superoxide dismutase?SOD?,malondialdehyde?MDA?,proline?Pro?,chlorophyll?Chlorophyll?.The results show the trend of rapid decline after reaching a certain degree.Under zinc stress,plant protein content,antioxidant capacity,anti-toxicity,and growth conditions all decline to varying degrees.This finding provides a certain reference basis for more physiological and biochemical studies of other plants under abiotic stress.2.Preliminary analysis of mulberry transcriptome sequencing and identification of differential genes:A total of 58,397 differential genes are obtained from the transcriptome sequencing experiments,of which 35,911 genes are annotated in the NR database,22,359 genes are annotated in the Swissprot database,20092 genes are annotated in the KOG database,and 13,689 genes are annotated in KEGG Database.Based on the results,the study performs alternative splicing prediction analysis,gene structure optimization analysis and the discovery of new genes,as well as gene expression analysis.Moreover,the study selects up and down-regulated genes for detection and analysis of relative expression levels.The results are consistent with the prediction,and the gene expression level also corresponds to the previous experimental results.The result is expected to lay a foundation for the follow-up gene research of mulberry under zinc stress.3.Cloning,sequence analysis and expression analysis of mulberry MalFLS gene:The complete ORF frame MalFLS gene is cloned in Fengchi Sang.The sequence length of the gene is 1077 bp,which contains the open reading frame of 1008 bp?start codon ATG,stop codon TAA?,the gene contains 335 amino acid encoding.It is predicted that the theoretical molecular weight Mw of the gene protein is 38.18 k D and the isoelectric point theory PI is 5.55,belonging to the 20G-FeII?0xy superfamily.4.Cloning,sequence analysis and expression analysis of mulberry MalCHS gene:The complete ORF frame MalCHS gene is cloned in Fengchi Sang.The sequence length of the gene is 1258 bp,which contained an open reading frame of1176 bp?start codon ATG,stop codon TAA?.the gene contains 389 amino acid encoding.It is predicted that the theoretical molecular weight Mw of the gene protein is 42.83 k D and the isoelectric point theory PI is 6.48,which belongs to the fabH superfamily.