| 朱晨璐,武欣怡,喻君保,黄偲祺,曹树一,翟林,韩雪洁,侯晓龙.基于转录组测序金丝草叶片响应铅胁迫的抗氧化酶相关基因[J].农业环境科学学报,2022,41(10):2158-2169. |
| 基于转录组测序金丝草叶片响应铅胁迫的抗氧化酶相关基因 |
| Antioxidant enzyme-related genes of Pogonatherum crinitum leaves in response to lead stress based on RNA-Seq |
| 投稿时间：2022-03-05 修订日期：2022-04-15 |
| DOI：10.11654/jaes.2022-0218 |
| 中文关键词: 超富集植物 金丝草 铅胁迫 抗氧化酶 实时荧光定量聚合酶链反应 |
| 英文关键词: hyperaccumulator Pogonatherum crinitum Pb stress antioxidase qRT-PCR |
| 基金项目:福建省科技厅对外合作项目(2019I0007);国家自然科学基金项目(31901298);福建农林大学科技创新专项基金(CXZX2020143C,71202103D);福建龙净环保股份有限公司项目(201906) |
| 作者 | 单位 | E-mail | | 朱晨璐 | 福建农林大学林学院, 福州 350002 | | | 武欣怡 | 福建农林大学林学院, 福州 350002 | | | 喻君保 | 福建农林大学林学院, 福州 350002 | | | 黄偲祺 | 福建农林大学林学院, 福州 350002 | | | 曹树一 | 福建农林大学林学院, 福州 350002 | | | 翟林 | 福建农林大学林学院, 福州 350002 | | | 韩雪洁 | 福建农林大学林学院, 福州 350002 | | | 侯晓龙 | 福建农林大学林学院, 福州 350002
南方红壤区水土保持国家林业和草原局重点实验室, 福州 350002
海峡两岸红壤 区水土保持协同创新中心, 福州 350002 | xl.hou@fafu.edu.cn |
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| 中文摘要: |
| 为了深入揭示植物抗氧化酶的相关基因及其响应重金属胁迫的作用机制，以重金属铅（Pb）超富集植物金丝草[Pogonatherum crinitum（Thunb.）Kunth.]为研究对象，设置Pb浓度0、300、500、1 000 mg·L-1和2 000 mg·L-1的水培模拟胁迫试验，测定不同Pb浓度下金丝草叶片的抗氧化酶活性，并对Pb胁迫处理的金丝草叶片进行RNA-Seq测序，将转录组测序得到的Unigenes通过GO与KEGG富集注释，筛选出叶片抗氧化酶相关的DEGs，并利用实时荧光定量聚合酶链反应（qRT-PCR）进行验证。结果表明：随Pb胁迫浓度的增加，金丝草叶片过氧化物酶（POD）、超氧化物歧化酶（SOD）和过氧化氢酶（CAT）活性呈增加趋势，丙二醛（MDA）含量则呈先增加后降低的趋势；转录组测序得到总碱基数为56.34 Gb，各样品碱基质量达到Q20、Q30水平的均大于90%，测序结果可靠；筛选出的DEGs通过GO与KEGG数据库进行对比注释发现，金丝草叶片DEGs在“过氧化物酶体”“氧化还原酶活性”“氧化还原酶活性，作用于CH-OH供体组”“活性氧代谢过程”和“过氧化氢代谢过程”等与抗氧化相关的GO功能条目上显著富集，在“谷胱甘肽代谢”“抗坏血酸和醛糖代谢”“植物激素信号转导”“黄酮类生物合成”和“MAPK信号通路-植物”等与抗氧化相关的KEGG通路上显著富集； Pb胁迫下金丝草叶片抗氧化酶相关基因PcSOD、PcPOD和PcCAT上调表达，qRT-PCR验证结果与转录组测序结果一致，Pb胁迫下这些基因的相对表达量与抗氧化酶活性的变化趋势一致。研究表明，金丝草可通过提高抗氧化酶活性来适应Pb胁迫，抗氧化酶相关基因PcSOD、PcPOD和PcCAT参与了金丝草应对Pb胁迫的调控过程。 |
| 英文摘要: |
| To further reveal the related genes of plant antioxidant enzymes and their mechanisms of response to heavy metal stress,Pogonatherum crinitum was used as the research object, and a hydroponic simulated stress experiment with lead concentrations of 0, 300, 500, 1 000 mg·L-1, and 2 000 mg·L-1 was carried out. In addition, the antioxidant enzyme activity in the leaves of P. crinitum at different concentrations was determined, and the sequencing of the RNA-Seq of P. crinitum leaves by Pb treatments was performed. The unigenes obtained by transcriptome sequencing were enriched and annotated with GO and KEGG, and the DEGs related to leaf antioxidant enzymes were screened out and verified using qRT-PCR. The results revealed that, with the increase in lead stress concentration, the activities of POD, SOD, and CAT in the leaves of P. crinitum showed an increasing trend; however, MDA content showed an increasing trend firstly and then decreased. The total number of bases obtained by sequencing was 56.34 Gb. The base quality of each sample reached Q20 and Q30 exceeded 90%, indicating that the database was reliable. The selected DEGs were compared and annotated through GO and KEGG databases, and the result showed that antioxidant-related GO functional terms such as“peroxisome” “oxidoreductase activity”and “oxidoreductase activity, acting on CH-OH group of donors” “reactive oxygen species metabolic process”and“hydrogen peroxide metabolic process”were significantly enriched in the DEGs of P. crinitum. Moreover, the DEGs were significantly enriched in antioxidantrelated KEGG pathways such as“glutathione metabolism” “ascorbate and aldarate metabolism” “plant hormone signal transduction” "flavonoid biosynthesis”and“MAPK signaling pathway-plant” . The expression of antioxidant enzyme-related genes PcSOD, PcPOD, and PcCAT in the leaves of P. crinitum were upregulated under Pb stress. The qRT-PCR results were consistent with the transcriptome sequencing results, and the relative expression levels of these genes were consistent with the changing trend of the antioxidant enzyme activities under Pb stress. This study shows that P. crinitum adapts to heavy metal Pb stress by increasing the activity of antioxidant enzymes, and the antioxidant enzyme-related genes PcSOD, PcPOD, and PcCAT are involved in the regulation of P. crinitum in response to heavy metal Pb stress. |
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