盐胁迫下不同抗性小麦幼苗生理生化特性及转录组分析

doi:10.13304/j.nykjdb.2022.0679

摘要/Abstract

摘要：

为探究不同抗盐性小麦对盐胁迫的响应差异，选用‘百农889’和‘中国春’小麦品种，利用200 mmol·L^-1 NaCl溶液处理离体叶片和小麦幼苗，对盐胁迫后小麦幼苗叶片的生理、生化特征进行分析，并基于转录组数据分析差异基因表达及可变剪切事件。结果表明，盐胁迫下，‘百农889’离体叶片的失绿程度较‘中国春’轻微；伊文思蓝染色后，‘中国春’离体叶片的蓝色着色范围更大，且细胞相对活性显著降低；‘中国春’幼苗表现出明显的萎蔫、倒伏，‘百农889’幼苗的形态与对照组无明显差异；2个品种的相对电解质渗漏率都有所增加，但‘中国春’幼苗的相对电解质渗漏率增幅更大；‘中国春’的净光合速率（net photosynthetic rate，P_n）和气孔导度（stomatal conductance，G_s）受到显著抑制，‘百农889’的净光合速率和气孔导度与对照比无明显变化。转录组测序结果分析表明，与‘中国春’相比，‘百农889’中上调差异表达基因2 299个，下调差异表达基因2 527个；对差异表达基因的GO（gene ontology）富集分析发现，与叶绿体、光合作用和质膜相关的功能基因被显著富集；KEGG（Kyoto encyclopedia of genes and genomes）富集分析发现，与植物激素信号转导和植物次生代谢物相关的通路被显著富集。盐胁迫下，‘中国春’表现出更多的可变剪切事件，且主要富集在RNA转运和内质网蛋白加工等通路中。因此，小麦可以通过调控与叶绿素、质膜、次生代谢物以及光合作用相关基因的表达来抵抗盐胁迫，可变剪切事件的发生也说明小麦可以通过调控基因互作网络来应对盐胁迫。

关键词: 小麦, 盐胁迫, 形态指标, 光合作用, 质膜完整性, 叶绿素, 转录组测序

Abstract:

To explore the response difference of different types of wheat to salt stress， the leaves of ‘Bainong 889’ and ‘Chinese spring’ seedlings were subjected in 200 mmol·L^-1 NaCl solution. The physiological and biochemical characteristics of wheat seedling under salt stress were analyzed， and the differences of gene expression and the occurrence of variable shearing events were investigated based on transcriptomics data. The results showed that， under salt stress， the leaves of ‘Bainong 889’ had less chlorosis than ‘Chinese spring’； the leaves of ‘Chinese spring’ stained by Evans blue were greater and the relative activity of cells was significantly reduced. In addition， ‘Chinese spring’ wheat seedlings significantly wilted under salt stress， while ‘Bainong 889’ was no significant difference compared with control in the morphology. The relative electrolyte leakage rates of both cultivars under salt stress were higher than the control， and that of ‘Chinese spring’ seedlings was the highest. Under salt stress， the concents of chlorophyll a， chlorophyll b and total chlorophyll in leaves of both ‘Bainong 889’ and ‘Chinese spring’ seedlings decreased， and the net photosynthetic rate （P_n） and stomatal conductance （G_s） of ‘Chinese spring’ seedlings were significantly inhibited， while those of ‘Bainong 889’ were similar to the control. Transcriptome sequencing results showed that 2 299 differentially expressed genes were up-regulated and 2 527 differentially expressed genes were down-regulated in ‘Bainong 889’ than ‘Chinese spring’. GO （gene ontology） enrichment analysis of differentially expressed genes suggested that functions related to chloroplast， photosynthesis and plasma membrane were significantly enriched. KEGG （Kyoto encyclopedia of genes and genomes） enrichment analysis showed that pathways related to plant hormone signal transduction and plant secondary metabolites were significantly enriched. The analysis of variable shear events in transcriptome data showed that there were more variable shear events in ‘Chinese spring’ under salt stress， which were mainly concentrated in RNA transport and protein processing in endoplasmic reticulum. Therefore， wheat could responded to salt stress by regulating the expression of genes related to chlorophyll， plasma membrane， secondary metabolites and photosynthesis. The occurrence of variable shear events also indicated that two types of varieties regulated the gene interaction network distinctively under salt stress.

Key words: wheat, salt stress, morphological index, photosynthesis, plasma membrane integrity, chlorophyll, transcriptome sequencing

中图分类号:

S512.1

李双, 王爱英, 焦浈, 池青, 孙昊, 焦涛. 盐胁迫下不同抗性小麦幼苗生理生化特性及转录组分析[J]. 中国农业科技导报, 2024, 26(2): 20-32.

Shuang LI, Aiying WANG, Zhen JIAO, Qing CHI, Hao SUN, Tao JIAO. Physiological and Chemical Characteristics and Transcriptome Analysis of Different Type of Wheat Seedlings Under Salt Stress[J]. Journal of Agricultural Science and Technology, 2024, 26(2): 20-32.

图/表 11

图1 盐胁迫下小麦离体叶片的形态和绿色强度A：叶片形态；B：叶片的绿色强度。不同小写字母表示不同处理之间在P<0.05水平差异显著

Fig. 1 Leaf morphology and geen intensity of isolated wheat leaf under salt stressA： Leaf morphology； B： Green intensity of leaf. Different lowercase letters indicate significant differences between different treatments at P<0.05 level

图2 盐胁迫下小麦离体叶片的伊文思蓝染色及细胞相对活性A：伊文思蓝染色；B：细胞相对活性。不同小写字母表示不同处理之间在P<0.05水平差异显著

Fig. 2 Evans blue staining and relative cell activity in isolated leaves of under salt stressA： Evans blue staining； B： Relative cell activity. Different lowercase letters indicate significant differences between different treatments at P<0.05 level

图3 盐胁迫下‘百农889’‘中国春’的植株形态和叶绿素含量

Fig. 3 Plant morphology and chlorophyll content of ‘Bainong 889’ and ‘Chinese spring’ under salt stress

图4 盐胁迫下‘百农889’‘中国春’叶片的光合特性注：不同小写字母表示不同处理之间在P<0.05水平差异显著。

Fig. 4 Photosynthetic characteristics of leaf of ‘Bainong 889’ and ‘Chinese spring’ under salt stressNote：Different lowercase letters indicate significant differences between different treatments at P<0.05 level.

图5 盐胁迫下‘百农889’‘中国春’叶片相对电解质渗透率Fig. 5　Relative electrolyte leakage rate of leaves in ‘Bainong 889’ and ‘Chinese spring’ under salt stress注：不同小写字母表示不同处理之间在P<0.05水平差异显著。Note：Different lowercase letters indicate significant differences between different treatments at P<0.05 level.

图6 盐胁迫下小麦叶片中基因的分布和差异表达A：基因分布情况的维恩图；B：盐胁迫下两个品种小麦差异表达基因表达量的散点图

Fig. 6 Gene distribution and differential expression in wheat leaves under salt stressA： Venn diagram of gene distribution； B： Scatter plot of differentially expressed genes in two wheat cultivars under salt stress

图7 盐胁迫下小麦叶片差异表达基因的GO富集（前10条）A：上调基因；B：下调基因

Fig. 7 GO enrichment map of DEGs in wheat leaves under salt stress （top 10）A： Up-regulated gene； B： Down-regulated gene

图8 盐胁迫下小麦叶片差异表达基因的KEGG通路富集（前10条）A：上调基因；B：下调基因

Fig. 8 KEGG pathway enrichment of differentially expressed genes in wheat leaves under salt stress （top 10）A： Up-regulated gene； B： Down-regulated gene

表1 转录组中可变剪切基因数/事件数统计

Table 1 Variable splicing gene number/event number statistics in transcriptome

可变剪切事件AS event	基因数/可变剪切数 Genes number/ASs number
可变剪切事件AS event	中国春 Chinese spring	百农889 Bainong 889
总计Total	8 966/14 307	8 880/14 173
SE	5 685/7 954	5 668/7 931
A5SS	1 312/1 476	1 286/1 445
A3SS	3 113/3 785	3 066/3 716
MXE	329/142	333/421
RI	603/680	583/660

图9 盐胁迫下‘中国春’和‘百农889’中特异和差异可变剪切A：盐胁迫下2个小麦品种中发生可变剪切基因的比较；B：百农889的差异可变剪切事件数量

Fig. 9 Specific and differential variable shear of ‘Chinese spring’ and ‘Bainong 889’ under salt stressA： Comparison of variable splicing genes in 2 wheat varieties under salt stress； B： Number of differential variable shearing events

图10 盐胁迫下中国春和百农889中差异可变剪切的富集分析A：GO富集分析；B：KEGG富集分析

Fig. 10 Enrichment analysis of differential variable shear in Chinese spring and Bainong 889 under salt stressA： GO enrichment analysis； B： KEGG enrichment analysis

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