硼胁迫下烟草叶片转录组分析

doi:10.13304/j.nykjdb.2021.0954

摘要/Abstract

摘要：

为揭示硼胁迫抑制烟叶生长的分子机理，以8叶期栽培烟草‘K326’为实验材料，分别用含硼量为0、0.05、1.00、2.00、3.00 mmol·L^-1的Hoagland培养液浇灌处理4周，而后检测烟叶中的总糖、蛋白质、叶绿素、H₂O₂、硼含量及抗氧化酶活性等，并通过转录组测序技术分析硼胁迫下烟叶差异表达基因及显著富集的代谢通路。结果显示，硼含量越高，对烟株的生长抑制越显著。2.00 mmol·L^-1的硼处理即可显著抑制烟草根、茎和叶生长；并显著抑制叶片中糖类物质代谢，增加H₂O₂含量及增强氧化物酶（POD）活性。与常规Hoagland培养液（含硼量0.05 mmol·L^-1）浇灌的烟草相比，仅响应2.00 mmol·L^-1硼胁迫的389个差异表达基因富集于糖代谢、脂类代谢、激素信号及植物抗病信号途径。说明高浓度硼胁迫可能影响烟叶中的碳代谢和脂类代谢，并可能通过改变激素信号途径抑制烟草生长。从而为深入理解烟草对硼胁迫的响应机制提供更多依据。

关键词: 烟草, 硼胁迫, 转录组, 代谢途径, 信号转导

Abstract:

The molecular mechanism of tobacco growth inhibition induced by boron stress was not clear yet. The tobacco ‘K326’ at 8 leaves stage as experimental materials were cultured in Hoagland solutions with 0， 0.05， 1.00， 2.00， 3.00 mmol·L^-1 boron， respectively. After 4 weeks of treatment， the contents of total sugar， protein， chlorophyll， H₂O₂， boron and the antioxidant enzyme activities in tobacco leaves were detected. The transcriptome sequencing technology was used to analyze the differentially expressed genes （DEGs） and the significantly enriched metabolic pathways responding to boron stress in tobacco leaves. The results showed that， the higher the boron content， the more significant inhibition were found on the growth of tobacco plants. 2.00 mmol·L^-1 boron treatment significantly inhibited the growth of roots， stems， and leaves of tobacco； and significantly inhibit carbohydrate metabolism， increase H₂O₂ contents， and enhance peroxidase （POD） activity in leaves. Compared with tobacco treated with normal Hoagland solution （0.05 mmol·L^-1 boron）， 389 DEGs that responded to 2.00 mmol·L^-1 boron stress specifically were enriched in sugar metabolism， lipid metabolism， hormone signaling， and plant disease resistance signaling pathways. It was found that the boron stress affected carbon and lipid metabolism in tobacco leaves， and inhibited tobacco growth by altering hormone signaling pathways. These results provided some basis for further understanding the mechanism of response to boron stress in tobacco.

Key words: tobacco, boron stress, transcriptome, metabolism pathway, signal transduction

中图分类号:

S572

王潇然, 李笑语, 孙慧, 于海东, 石永春. 硼胁迫下烟草叶片转录组分析[J]. 中国农业科技导报, 2023, 25(8): 53-64.

Xiaoran WANG, Xiaoyu LI, Hui SUN, Haidong YU, Yongchun SHI. Transcriptome Analysis of Tobacco Leaves Under Boron Stress[J]. Journal of Agricultural Science and Technology, 2023, 25(8): 53-64.

图/表 12

参考文献 41

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基因编号 Gene ID	基因描述 Gene description	正向引物 Forward primer （5’-3’）	反向引物 Reverse primer （5’-3’）	产物长度Product size/bp
EU938079	肌动蛋白Actin	GGAAACATCGTCCTTAGTGGTG	AATCCAGACACTGAACTTGCGT	141
LOC104239595	蛋白TIFY8 Protein TIFY 8	TAATGTCCATCCCAACAAGGC	TTTAGGCGAACTATTTTCTCCAG	127
LOC107823411	碱性 β-1，3-葡聚糖酶 Basic β-1，3-glucanase	ACTGGCACATCTTACCTAACCTC	CCAACAATGGGATCAACAAAC	194
LOC107826794	内切几丁质酶B Endochitinase B	CTGCTGGCTTAGAGAACAAGGTAG	GCTCTTCCACAAGGTCCGTAG	141
LOC104119660	乙酰辅酶A氧化酶2 Acyl-coenzyme A oxidase 2	CTTGGAGGATTTGGTGCTTG	GTTTTAGGGCAGCACGAGAG	142
LOC104094164	乌头酸水合酶 Aconitate hydratase	GCAAAGTATCTTATGGAGCGTG	CCCAGTCGGTATGTGAATCG	168
LOC104087505	FK506结合蛋白类似物异构体X2 FK506-binding protein 5-like isoform X2	CAAAGGAAGGCAGACGGTG	GCGTCAAGAATCCCTGCTC	125
LOC104242999	细胞色素P450 CYP72A219类似物 Cytochrome P450 CYP72A219 like	CATTGCCAGCAGGGATACAG	GCCATTTTTGCCTCTAACATTG	211
LOC104236587	α，α-海藻糖-磷酸合成酶11异构体 X2 α，α-trehalose-phosphate synthase 11 isoform X2	GTCTTGGCTTCAGAGTTGTCG	CACTCGGGGTTTTGTCCAC	156

基因编号 Gene ID	基因描述 Gene description	正向引物 Forward primer （5’-3’）	反向引物 Reverse primer （5’-3’）	产物长度Product size/bp
EU938079	肌动蛋白Actin	GGAAACATCGTCCTTAGTGGTG	AATCCAGACACTGAACTTGCGT	141
LOC104239595	蛋白TIFY8 Protein TIFY 8	TAATGTCCATCCCAACAAGGC	TTTAGGCGAACTATTTTCTCCAG	127
LOC107823411	碱性 β-1，3-葡聚糖酶 Basic β-1，3-glucanase	ACTGGCACATCTTACCTAACCTC	CCAACAATGGGATCAACAAAC	194
LOC107826794	内切几丁质酶B Endochitinase B	CTGCTGGCTTAGAGAACAAGGTAG	GCTCTTCCACAAGGTCCGTAG	141
LOC104119660	乙酰辅酶A氧化酶2 Acyl-coenzyme A oxidase 2	CTTGGAGGATTTGGTGCTTG	GTTTTAGGGCAGCACGAGAG	142
LOC104094164	乌头酸水合酶 Aconitate hydratase	GCAAAGTATCTTATGGAGCGTG	CCCAGTCGGTATGTGAATCG	168
LOC104087505	FK506结合蛋白类似物异构体X2 FK506-binding protein 5-like isoform X2	CAAAGGAAGGCAGACGGTG	GCGTCAAGAATCCCTGCTC	125
LOC104242999	细胞色素P450 CYP72A219类似物 Cytochrome P450 CYP72A219 like	CATTGCCAGCAGGGATACAG	GCCATTTTTGCCTCTAACATTG	211
LOC104236587	α，α-海藻糖-磷酸合成酶11异构体 X2 α，α-trehalose-phosphate synthase 11 isoform X2	GTCTTGGCTTCAGAGTTGTCG	CACTCGGGGTTTTGTCCAC	156

器官Organ	T₀	T₂
叶Leaf	0.899±0.065 Aa	6.523±0.135 Aa
茎Stem	0.869±0.026 Aa	1.176±0.107 Ab
根Root	0.570±0.059 Bb	0.886±0.011 Ab

器官Organ	T₀	T₂
叶Leaf	0.899±0.065 Aa	6.523±0.135 Aa
茎Stem	0.869±0.026 Aa	1.176±0.107 Ab
根Root	0.570±0.059 Bb	0.886±0.011 Ab

性状Trait	T₀	T₂
可溶性糖含量Soluble sugar content	8.165±0.167 Aa	4.711±0.109 Bb
葡萄糖含量Glucose content	0.864±0.032 Aa	0.871±0.051 Aa
果糖含量Fructose content	1.526±0.043 Aa	1.755±0.028 Bb
蔗糖含量Sucrose content	4.726±0.031 Aa	1.815±0.019 Bb
淀粉含量Starch content	46.551±0.143 Aa	22.653±0.127 Bb
叶绿素a含量Chlorophyll a content	1.746±0.028 Aa	1.632±0.019 Aa
叶绿素b含量Chlorophyll b content	0.800±0.009 Aa	0.741±0.013 Aa
类胡萝卜素含量Carotenoids content	0.295±0.025 Aa	0.265±0.008 Aa
纤维素含量Cellulose content	20.685±0.236 Aa	13.24±0.433 Bb
果胶含量Pectin content	46.313±0.563 Aa	30.500±1.250 Bb
脯氨酸含量Proline content	15.477±0.349 Aa	24.488±0.988 Bb
蛋白质含量Protein content	47.836±2.506 Aa	67.540±1.481 Bb
过氧化氢含量H₂O₂ content	0.450±0.120 Aa	2.339±0.096 Bb