| Phospholipase is a class of enzymes that catalyze the hydrolysis of glycerophospholipids.According to the different substrates,it can be divided into four categories,namely Phospholipase A（Phospholipase A,PLA）,Phospholipase B（Phospholipase B,PLB）,Phospholipase C（Phospholipase C,PLC）,Phospholipase D（Phospholipase D,PLD）four categories.In plants,phospholipase C can be further subdivided into phosphoinositide-specific PLC（PI-PLC）and phosphatidylcholine phospholipase C（phosphatidylcholine-PLC,PC-PLC）.In this study,the maize inbred line He 344 was used as the test material,Neomycin Sulphate（NS）was selected as the inhibitor,and 100 m M NS was used to inhibit the maize seedlings with two leaves and one heart.Using high-throughput transcriptome sequencing and lipidome detection technology,analyze the regulation of lipid metabolism and the differential expression of phospholipid metabolism genes in maize seedling leaves under the inhibition of phospholipase C,and explore phospholipids through molecular,physiological and biochemical aspects How does enzyme C participate in photosynthetic response and lipid metabolism regulation.Provide a theoretical basis for discovering the function of phospholipase C.The main results of the research are as follows:1.Under the inhibition of phospholipase C,the leaves of maize seedlings gradually become green and red,and the leaves gradually wilt.Compared with the control,the plants are shorter.The plant height,leaf fresh weight,leaf dry weight,root length,root fresh weight and root dry weight all increased with the increase of treatment time,but they were all less than the control.At the same time,the number of branches,the number of root tips,the diameter of the root system and the total length of the root system increased with the increase of treatment time,but the treatment was less than that of the control.The above results indicate that phospholipase C is involved in the growth and development of the lateral roots of maize seedlings.The leaves were stained with E-vans blue,and it was found that after the treatment,the degree of leaf coloration gradually increased with the increase of the treatment time.The color was the darkest in treatment 7D,indicating that the maize leaf cell damage was the most serious when 7D was inhibited by phospholipase C.The results of MAD determination are consistent.This shows that the inhibition of phospholipase C affects the growth and development of maize seedlings and destroys the stability of cell membranes.2.Reduce photosynthesis,photosynthesis and the factors that affect photosynthesis to supplement.With the increase of time,the maximum photosynthetic speed with synchronization gradually decreases,and the dark suppression effect decreases.With the increase of time,the increase of sugar,the increase time increases the amplitude control of 3d,with the increase of time,the increase of sugar is almost the same as the processing time.Decrease trend.Studies on photosynthetic growth and stomata have found that from biological effects to biochemical effects on the inhibition of C,the expression related to the synthesis of chlorophyll heads leads to the reduction of chlorophyll head synthesis precursors,which ultimately affects the reduction of chlorophyll,and ultimately affects the synthesis of chlorophyll:On the hole,the hole conductance is mainly synthesized,which reduces the entry.As a result,air element affects CO2stomatal enzyme C to participate in green synthesis and pore opening and closing,thereby achieving the photosynthesis of maize seedlings..3.In this study,the BMK sequencing platform was used to sequence the transcriptome of maize leaves that were inhibited with 0d,1d,3d,and 5d.The results show that the 5d NS vs0d CK comparison group has the largest number of differential genes,indicating that 5d under phospholipase C inhibition will induce a large number of phospholipid-responsive gene expression.Compare and annotate the differentially expressed genes whose expression level changes by more than 1（|Log2FC|≥1）in the comparison group with the transcription factor database,and it is found that the NAC and b HLH transcription factor families have the largest number of differentially expressed genes.Through the correlation analysis of the expression of the phospholipase C gene,it was found that the PLCs of maize showed a certain correlation in the expression.It is found that Zm PI-PLC3 and Zm NPC4 show a significant positive correlation,with a correlation coefficient of 0.99;Zm PI-PLC2 and Zm NPC2 show a significant negative correlation,with a correlation coefficient close to-1.4.The lipid regulatory genes selected from the transcriptome data are significantly enriched in differential genes involved in the metabolic pathways of"phosphatidylinositol signal system"and"phytohormone signal transduction".At the same time,MGDG,DGDG and PC are significantly different.At the same time,the content of DAG has a certain correlation with MGDG and PI.Under inhibiting conditions,the PIP2 degradation pathway and the PC/NPC pathway are inhibited,which leads to a decrease in the amount of DAG and IP3 produced on the cell membrane and a decrease in the amount of DAG produced through the PC/NPC pathway.We found that with the increase of phospholipase C inhibition time,the PAH and PLD/PAP/LPP pathways are activated,which leads to an increase in DAG content in the late stage of inhibition.These findings indicate that under the inhibition of phospholipase C,the turnover rate of DAG is increased,which explains the increase in DGA content and the accumulation of PC in the later stage of inhibition revealed by lipidomics analysis.Based on the results of transcriptome,physiological and lipidome data,this study initially established a gene-metabolite interaction network diagram of membrane lipid metabolism in maize leaves,which provides a theoretical basis for in-depth analysis of the role of phospholipase C in the phospholipid signaling pathway. |
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