不同浓度K^(+)处理下枸杞K^(+)通道相关基因表达水平分析

Differential expression levels of K^(+)channel related genes in Lycium Barbarum under different concentrations of K^(+)treatment

【目的】利用不同浓度的KCl溶液对‘青杞1号’的组培幼苗进行处理,对处理后的根部组织进行实验研究,研究并分析与钾离子通道相关的基因在宁夏枸杞中的表达模式。【方法】对不同浓度KCl处理后的宁夏枸杞幼苗根部进行RNA提取,采用实时荧光定量PCR等方法,分析在不同浓度钾离子处理条件下,枸杞中钾离子通道相关基因表达量的变化情况。【结果】随着KCl处理浓度的增加,‘青杞1号’中KCO1B、TPK1、KT1、Cluster-17443.10870、Cluster-17443.69422和Cluster-17443.15033基因总体都为先上升再下降最后上升的趋势,其中,Cluster-17443.10870基因在K^(+)处理浓度变化至100 mmol/L时有一定程度的下降。KCO1B基因在150 mmol/L时表达量达到最大;KT1基因和Cluster-17443.69422基因在200 mmol/L时表达量达到最大;TPK1基因、Cluster-17443.10870基因和Cluster-17443.15033基因在350 mmol/L时表达量才达到最大。350 mmol/L K^(+)处理条件下各组基因的表达量都出现突然上升状态,预测此时钾通道活性已达到过饱和状态;在100 mmol/L K^(+)处理时,有多数基因的表达量低于对照组,预测此浓度下钾通道功能受抑制。其他浓度下各组基因的表达量都在中间处理浓度(150、200、250 mmol/L)时达到最大值,在最低(100 mmol/L)或最高(300 mmol/L)浓度处理时基因的表达量达到最小值。【结论】盐胁迫浓度的增加使宁夏枸杞钾离子通道相关基因的表达量总体呈现出先升后降再升和先升后降的变化趋势。本研究为今后研究枸杞钾盐胁迫调节机制提供了部分理论基础,为柴达木地区枸杞产业提质增效和盐碱地造林等工作提供理论支持,对促进绿色化、生态化枸杞种植产业的发展具有潜在意义。

【Objective】The tissue culture seedlings of Qingqi No.1 were treated with different concentrations of KCl solutions.The root tissues after the treatment were experimentally studied,and the expression patterns of genes related to potassium channels in Lycium barbarum were studied and analyzed.【Method】RNA from the roots of Lycium barbarum seedlings treated with different concentrations of KCl was extracted.Using qRT-PCR and other methods,the changes in the expression of potassium channelrelated genes in Lycium barbarum under different potassium ion treatments were analyzed.【Result】With the increase of KCl concentration,the gene expression of KCO1B,TPK1,KT1,Cluster-17443.10870,Cluster-17443.69422 and Cluster-17443.15033 in Qingqi No.1 all increased first,then decreased and finally increased.Among them,Cluster-17443.10870 decreased to a certain extent when the K^(+)concentration was changed to 100 mmol/L.The expression level of KCO1B reached the maximum level at 150 mmol/L;the expression level of KT1 and Cluster-17443.69422 reached the maximum level at 200 mmol/L;TPK1,Cluster-17443.10870 and Cluster-17443.15033 showed the highest expression levels when the concentration of K^(+)was 350 mmol/L.Under the condition of 350mmol/L K^(+)treatment,the expression of all detected genes exhibited a sudden increase.It was predicted that the activity of the potassium channel had reached an over-saturated state at this time.With a K^(+)concentration of 100 mmol/L,the expression of most genes was lower than those of the control group,suggesting the functional inhibition of potassium channel.At other concentrations(150,200,250 mmol/L),the expression of all detected genes reached the maximum values at the intermediate concentration,while among all treatments,the genes always showed the lowest expression levels at the lowest(100 mmol/L)or highest(300 mmol/L)concentration.【Conclusion】With the increase of salt concentration,the expression of potassium channel-related genes of Lycium barbarum shows a trend of first increase,then decrease and increase again,or an increase followed by a decrease.This study provides a theoretical basis for the research on the regulation mechanism of wolfberry potassium responding to salt stress,and theoretical support for the improvement of the wolfberry industry and the afforestation of saline-alkali land in the Qaidam area.It is of great significance to promote green and ecological wolfberry planting.