Transcriptome Profiling and Gene Mining of Millet Response to Potassium Deficiency Stress

doi:10.13304/j.nykjdb.2023.0900

Abstract

Abstract:

Potassium is an essential element for crop growth. It is of great significance to explore genes that improve crop potassium absorption capacity and enhance crop tolerance to potassium deficiency. In this study， ‘Jigu 45’ was used as material and its seedlings were subjected to potassium deficiency stress for 7 d at the 6-leaf stage. By analyzing agronomic indicators and transcriptome， the relevant genes and expression regulation of potassium deficiency stress were analyzed and explored. The results showed that potassium deficiency stress affected the growth and development of foxtail millet plants： the plant height， leaf width， leaf length， stem width， aboveground dry weight and chlorophyll content were lower then those of CK， while the root length and root surface area significantly increased. Transcriptome analysis showed that 217 genes were upregulated and 38 genes were downregulated after potassium deficiency stress. GO functional enrichment revealed that differentially expressed genes were mainly enriched in the structural components of ribosomes， organic nitrogen compound metabolism processes， protein metabolism processes， cellular protein metabolism processes， and membraneless organelles etc. The KEGG functional enrichment results determined that differentially expressed genes were mainly enriched in 19 pathways， including ribosomes， plant hormone signal transduction and glutathione metabolism etc. Subsequently， 35 genes significantly correlated with hormone expression， 19 differentially expressed genes related to oxidative stress， 10 transcription factor families， and 11 genes related to transduction signals were screened， as well as 1 gene related to sugar metabolism of amino acids and nucleotides. The candidate gene Seita.9G193900 belonged to the CYP45084A subfamily and was a key enzyme regulating S-type lignin synthesis； Seita.5G36500 belonged to WRKY family， which regulated the expression of disease response genes such as PAD3； Seita.3G216900 was an RNA polymeraseⅡtranscription subunit 37E related mediator， HSPA1s helper protein； Seita.4G286000 was CHB gene which coded chitin endonuclease that regulated sugar synthesis and fermentation. Above results revealed the transcriptional regulation of genes related to potassium deficiency stress in foxtail millet， and laid a foundation for the cloning and functional verification of genes related to potassium tolerance in foxtail millet.

Key words: millet, potassium deficiency, differentially expressed genes, transcriptome

摘要：

钾是作物生长不可缺少的元素，挖掘提高作物钾吸收能力的基因，增强作物对缺钾的耐受性具有重要的意义。以‘冀谷45’为供试材料，幼苗长至6叶期时进行缺钾胁迫7 d，通过农艺指标及转录组分析挖掘缺钾胁迫的相关基因。结果表明，缺钾胁迫影响谷子的生长发育，株高、叶宽、叶长、茎粗、苗期地上部干重及叶绿素含量低于CK，而根长、根表面积显著增加。转录组分析显示，缺钾胁迫后217个基因表达上调，38个基因表达下调。GO功能富集发现，差异表达基因主要富集于核糖体的结构成分、有机氮化合物代谢过程、蛋白质代谢过程、细胞蛋白质代谢过程、无膜细胞器等类别。KEGG功能富集结果表明，差异表达基因主要富集于核糖体、植物激素信号转导、谷胱甘肽代谢等19个通路。进而筛选出35个与激素表达显著相关的基因，19个氧化应激相关差异表达基因，10个转录因子家族及11个与转导信号相关的基因，1个与氨基酸及核苷酸的糖代谢相关的基因。候选基因Seita.9G193900属于CYP45084A亚家族，调控S型木质素合成的关键酶；Seita.5G365500属于WRKY家族，调控抗病响应基因PAD3等基因的表达；Seita.3G216900是RNA聚合酶Ⅱ转录亚基37E相关介质——HSPA1s辅助蛋白；Seita.4G286000为几丁内质内切酶基因CHIB，调控糖的合成与酵解。以上结果初步揭示了谷子缺钾胁迫相关基因的转录调控，为谷子耐缺钾相关基因的克隆与功能验证奠定基础。

关键词: 谷子, 缺钾, 差异表达基因, 转录组

CLC Number:

S515

Yiwei LU, Xueyan XIA, Yu ZHAO, Jihan CUI, Meng LIU, Meihong HUANG, Cheng CHU, Jianjun LIU, Shunguo LI. Transcriptome Profiling and Gene Mining of Millet Response to Potassium Deficiency Stress[J]. Journal of Agricultural Science and Technology, 2024, 26(6): 30-44.

鲁一薇, 夏雪岩, 赵宇, 崔纪菡, 刘猛, 黄玫红, 褚程, 刘建军, 李顺国. 缺钾胁迫下谷子转录组分析及相关基因挖掘[J]. 中国农业科技导报, 2024, 26(6): 30-44.

Figures/Tables 16

References 36

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成分Component	对照 Control	缺钾 Potassium deficiency
1.0 mol·L^-1 Ca（NO₃）₂·4H₂O	5	5
1.0 mol·L^-1 KNO₃	5	—
1 mol·L^-1 NaH₂PO₄	—	1
1.0 mol·L^-1 NaNO₃	—	5
1.0 mol·L^-1 MgSO₄·7H₂O	2	2
1.0 mol·L^-1 KH₂PO₄	1	—
0.5% Fe-EDTA	1	1
微量元素母液 Trace element mother liquor	1	1
ddH₂O	985	985
总体积 Total volume	1 000	1 000

成分Component	对照 Control	缺钾 Potassium deficiency
1.0 mol·L^-1 Ca（NO₃）₂·4H₂O	5	5
1.0 mol·L^-1 KNO₃	5	—
1 mol·L^-1 NaH₂PO₄	—	1
1.0 mol·L^-1 NaNO₃	—	5
1.0 mol·L^-1 MgSO₄·7H₂O	2	2
1.0 mol·L^-1 KH₂PO₄	1	—
0.5% Fe-EDTA	1	1
微量元素母液 Trace element mother liquor	1	1
ddH₂O	985	985
总体积 Total volume	1 000	1 000

引物名称 Primer name	引物序列 Primer sequence （5'-3 '）
Seita.9G193900-F2	ATGTTTGGCGGGACGGAGAC
Seita.9G193900-R2	CCGATGGCCCAGACGTTGA
Seita.5G299700-F	GCGGGAGGACTTCGTGATGC
Seita.5G299700-R	CGGTGACGGGAATGCTGGA
Seita.5G365500-F1	CGGCCCTCCTCACCTCCAGTAT
Seita.5G365500-R1	CGCGTCCATGCCTTGTTGC
Seita.3G216900-F	ACCCGTCCGTGCAGAGTGA
Seita.3G216900-R	GGCAGTGGGCTCGTTGATT
Seita.7G222200-F1	GGTGGTGGAGGTCTTCGTC
Seita.7G222200-R1	TGTTGCCACTGTGCTTGATG
Seita.4G286000-F2	GACGCCACCATCGCCTTCA
Seita.4G286000-R2	GTTGGACCCGTAGCTGACCC
Seita.2G285600-F	TGCACGTCCACTGCCACATC
Seita.2G285600-R	CACGAACGCCTTCAGAACCAC
SiActin-F	GGCAAACAGGGAGAAGATGA
SiActin-R	GAGGTTGTCGGTAAGGTCACG

引物名称 Primer name	引物序列 Primer sequence （5'-3 '）
Seita.9G193900-F2	ATGTTTGGCGGGACGGAGAC
Seita.9G193900-R2	CCGATGGCCCAGACGTTGA
Seita.5G299700-F	GCGGGAGGACTTCGTGATGC
Seita.5G299700-R	CGGTGACGGGAATGCTGGA
Seita.5G365500-F1	CGGCCCTCCTCACCTCCAGTAT
Seita.5G365500-R1	CGCGTCCATGCCTTGTTGC
Seita.3G216900-F	ACCCGTCCGTGCAGAGTGA
Seita.3G216900-R	GGCAGTGGGCTCGTTGATT
Seita.7G222200-F1	GGTGGTGGAGGTCTTCGTC
Seita.7G222200-R1	TGTTGCCACTGTGCTTGATG
Seita.4G286000-F2	GACGCCACCATCGCCTTCA
Seita.4G286000-R2	GTTGGACCCGTAGCTGACCC
Seita.2G285600-F	TGCACGTCCACTGCCACATC
Seita.2G285600-R	CACGAACGCCTTCAGAACCAC
SiActin-F	GGCAAACAGGGAGAAGATGA
SiActin-R	GAGGTTGTCGGTAAGGTCACG

性状 Trait	对照 Control	缺钾 Potassium deficiency
株高 Plant height /cm	12.43	9.36^*
倒一叶叶宽 Top 1 leaf width /cm	0.80	0.67
倒一叶叶长 Top 1 leaf length /cm	20.75	17.75^*
倒二叶叶宽 Top 2 leaf width /cm	0.80	0.67
倒二叶叶长 Top 2 leaf length/ cm	19.63	19.50
倒三叶叶宽 Top 3 leaf width/cm	0.80	0.60
倒三叶叶长 Top 3 leaf length /cm	18.05	16.55^*
茎宽 Stem width /cm	3.31	2.51
苗期地上部干重 Seedling stage aboveground dry weight/g	0.07	0.06
苗期地下部干重 Seedling stage underground dry weight /g	0.01	0.02