镉胁迫下美洲黑杨无性系‘中菏1号’转录组分析

宋润先; 李翔; 毛秀红; 王丽; 陈香丽; 姚俊修; 赵曦阳; 李善文

doi:10.12171/j.1000-1522.20210037

镉胁迫下美洲黑杨无性系‘中菏1号’转录组分析

Transcriptome analysis of clone Populus deltoides ‘Zhonghe 1’ under cadmium stress

摘要

摘要:
目的通过转录组测序和分析，发掘美洲黑杨无性系‘中菏1号’耐镉关键功能基因，揭示美洲黑杨在镉胁迫下的响应机制。
方法以‘中菏1号’为试验材料，通过沙培试验法对处在不同质量浓度（0 mg/L（ZH1C0），10 mg/L（ZH1C1），20 mg/L（ZH1C2））Cd²⁺ 环境下的无性系进行胁迫处理，而后采集叶片，提取RNA，并使用Illumina HiSeq^TM2500系统对转录组进行高通量测序。
结果主成分分析确定PC1作为区分不同Cd²⁺ 浓度样本的标准；经过差异表达分析，在ZH1C0和ZH1C2之间共检测到4 109条差异基因，其中2 741条基因显著上调表达，1 368条基因显著下调表达；在ZH1C1和ZH1C2之间检测到3 710条差异基因，其中1 969条基因显著上调表达，1 741条基因显著下调表达；不同数据库之间的注释表明Cd²⁺ 不仅会影响叶绿体、线粒体等细胞器的微结构，而且会干扰光合作用和呼吸作用中的电子传递。转录因子分析发现，WRKY、MYB、ZIP、ERF、bHLH在镉胁迫响应和耐受中具有重要作用。
结论在‘中菏1号’中，异戊二烯化植物蛋白在阻止Cd²⁺ 进入细胞内部发挥了重要作用；谷胱甘肽不仅可以促进细胞内Cd²⁺ 的螯合，还可以缓解细胞内的氧化胁迫；ABC家族蛋白和MATE家族蛋白是‘中菏1号’体内转移Cd²⁺ 螯合蛋白的关键载体。

Abstract:
Objective Our objective was to explore the molecular mechanism of cadmium stress in Populus deltoides ‘Zhonghe 1’, search for anti-cadmium genes by transcriptome analysis and provide candidate plants for phytoremediation.
Method The experiment used sand to cultivate the P. deltoides ‘Zhonghe 1’. Different mass concentrations (0 mg/L (ZH1C0), 10 mg/L (ZH1C1), 20 mg/L (ZH1C2)) of cadmium ionic liquor were used to stress the clones. Then RNA extracted from leaves was sequenced by the system Illumina HiSeq^TM2500.
Result PC1 is the principal component which can distinguish samples at different concentrations. The comparison between ZH1C0 and ZH1C2 in differential expression analysis revealed that 4 109 differential genes were detected, among which 2 741 were up-regulated and 1 368 were down-regulated. Similarly, in comparison with the ZH1C1 and ZH1C2 in differential expression analysis, 3 710 differential genes were detected with 1 969 up-regulated genes and 1 741 down-regulated ones. It was found by the annotation, which were in different databases that cadmium stress could affect the electron transport in chloroplasts and mitochondria. Transcription factor analysis showed that WRKY, MYB, ZIP, bHLH and ERF played an important role in cadmium stress.
Conclusion The results indicate that HIPPs play a crucial part in preventing the Cd²⁺ from entering the cell. GSH could not only promote the Cd²⁺ binding but also alleviate intracellular oxidative stress. The ABC family proteins and the MATE family proteins are the key carriers for the transfer of the Cd-PCs in the P. deltoides ‘Zhonghe 1’.

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