Analysis of the key exogenous IAA-induced gene expression levels and metabolic pathways involved in long-distance translocation of Cd in Poa pratensis

doi:10.11686/cyxb2022276

Abstract

Abstract:

This research aimed to investigate the effects and molecular mechanisms of indole-3-acetic acid （IAA）-induced decrease of the long-distance translocation of cadmium （Cd） from the roots to the shoots in Kentucky bluegrass by analysis the differentially expressed IAA-induced genes （DEGs） and metabolic pathways involved in response to Cd stress. The species Poa pratensis was used as the study material and 400 nmol·L^-1 IAA was sprayed on the leaves of plants subjected to 600 μmol·L^-1 of Cd stress. Plant heights， root lengths， and Cd concentrations in the roots and leaves were measured， and transcriptome sequencing was performed in both the Cd+ IAA and Cd treatments. Real-time quantitative polymerase chain reaction （RT-qPCR） was used to verify the accuracy of the RNA-seq. It was found that exogenous IAA significantly increased the plant heights and root lengths， and inhibited the long-distance translocation of Cd from roots to leaves. There were 1294 genes with significantly IAA-modified expression under Cd stress in Kentucky bluegrass. The expression levels of RT-qPCR had a high correlation with RNA-seq data. Gene ontology （GO） enrichment analysis showed that the genes involved in response to heat， cold， Cd， and virus exposure were induced by IAA. The molecular chaperone HSP70， protein kinase mitogen-activated protein kinase （MAPK）， and transcription factor myeloblastosis46 （MYB46） were very obviously up-regulated by IAA application. In addition， genes involved in the glutathione metabolism and lignin biosynthesis pathways were also up-regulated by exogenous IAA. This may be indicative of increased glutathione synthesis and lignification of the cell wall and further increase the vacuolar compartmentalization and decreased the Cd entry into cells， and further decrease the Cd concentration in leaves. This study provides reference data on mechanisms of IAA-induced translocation and distribution of Cd.

Key words: Cd stress, Kentucky bluegrass （Poa pratensis）, auxin, transcriptome, translocation of Cd

Ting CUI, Yong WANG, Hui-ling MA. Analysis of the key exogenous IAA-induced gene expression levels and metabolic pathways involved in long-distance translocation of Cd in Poa pratensis[J]. Acta Prataculturae Sinica, 2023, 32(6): 146-156.

Figures/Tables 9

References 36

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基因ID Gene ID	上游引物Forward primers	下游引物Reverse primers
TRINITY_DN115423_c2_g1	GCACATGTTCGTCGTCAAGG	AATCTCCTTCTTGTCCGGCG
TRINITY_DN145731_c0_g2	AAGAGCCATTGCCGTACTCC	TGGTCGTGCTTGGTCTCAAA
TRINITY_DN163800_c1_g2	TCTCGAGATCTGACCTGGCA	GTTCGTCGCCACTTGGAGTA
TRINITY_DN145701_c0_g1	CATCACGTCTGTGAACCCCA	GTCCGTGTTTGGGCTGTTTC
TRINITY_DN141021_c0_g2	CAACACATTCCCACCCTCCA	CAGCTGGATGTTCACCGGAT
TRINITY_DN142220_c1_g2	GACTGGCCAACTCAGACCTC	CGACCGTGACCTTCATAGGG
TRINITY_DN135644_c0_g2	TTCTACCGTTGCAGCTTCGT	TGGTGCCGTAGATGTCACAC
TRINITY_DN155113_c1_g1	CTTCACTCACTGCCAAACGC	GAGGCAAAAGACTGCTGCAC
TRINITY_DN72438_c0_g1	GACCATTGTCACCGGTCGTA	ATCTCCCTGGTTGCGGAATG
TRINITY_DN168047_c0_g1	ATCCTAGCGCCTTCATGCTC	GTACTTGCCGACACCAGTGA
TRINITY_DN139972_c1_g1	TGGCAGGCCCTACCTAGATT	TCCCAAAGCTCCTGCTTGTC
TRINITY_DN167060_c4_g1	AGGGCCGCTACTAAATACGC	CAGCACCTGAGCTTTCCTGA
TRINITY_DN163988_c1_g1	TGCAATATCCCAAGCAGCCA	CAGTCTTTAGCCCCTCACCG
TRINITY_DN165172_c2_g1	CAGCAGGCCCTGTCATTGTA	CGGTACCGGAACACGAAGAA
TRINITY_DN165077_c2_g1	CGGCGATTCTCTACCTCGTC	GCGTGGAATCGTTCTTGAGC

基因ID Gene ID	上游引物Forward primers	下游引物Reverse primers
TRINITY_DN115423_c2_g1	GCACATGTTCGTCGTCAAGG	AATCTCCTTCTTGTCCGGCG
TRINITY_DN145731_c0_g2	AAGAGCCATTGCCGTACTCC	TGGTCGTGCTTGGTCTCAAA
TRINITY_DN163800_c1_g2	TCTCGAGATCTGACCTGGCA	GTTCGTCGCCACTTGGAGTA
TRINITY_DN145701_c0_g1	CATCACGTCTGTGAACCCCA	GTCCGTGTTTGGGCTGTTTC
TRINITY_DN141021_c0_g2	CAACACATTCCCACCCTCCA	CAGCTGGATGTTCACCGGAT
TRINITY_DN142220_c1_g2	GACTGGCCAACTCAGACCTC	CGACCGTGACCTTCATAGGG
TRINITY_DN135644_c0_g2	TTCTACCGTTGCAGCTTCGT	TGGTGCCGTAGATGTCACAC
TRINITY_DN155113_c1_g1	CTTCACTCACTGCCAAACGC	GAGGCAAAAGACTGCTGCAC
TRINITY_DN72438_c0_g1	GACCATTGTCACCGGTCGTA	ATCTCCCTGGTTGCGGAATG
TRINITY_DN168047_c0_g1	ATCCTAGCGCCTTCATGCTC	GTACTTGCCGACACCAGTGA
TRINITY_DN139972_c1_g1	TGGCAGGCCCTACCTAGATT	TCCCAAAGCTCCTGCTTGTC
TRINITY_DN167060_c4_g1	AGGGCCGCTACTAAATACGC	CAGCACCTGAGCTTTCCTGA
TRINITY_DN163988_c1_g1	TGCAATATCCCAAGCAGCCA	CAGTCTTTAGCCCCTCACCG
TRINITY_DN165172_c2_g1	CAGCAGGCCCTGTCATTGTA	CGGTACCGGAACACGAAGAA
TRINITY_DN165077_c2_g1	CGGCGATTCTCTACCTCGTC	GCGTGGAATCGTTCTTGAGC

样本Simples	原始序列数量Raw reads	清洁序列数量Valid reads	Q₂₀ （%）	Q₃₀ （%）	GC （%）
Cd_1	47397864	46440412	98.46	94.79	52.51
Cd_2	50450066	49185876	98.54	95.08	52.75
Cd_3	51288708	50076930	98.49	94.94	52.78
Cd+IAA_1	47775358	46702260	98.51	94.97	52.66
Cd+IAA_2	52741792	51346028	98.46	94.89	52.86
Cd+IAA_3	54986698	53830218	98.53	95.05	52.72
CK_1	52312058	51101542	98.48	94.87	52.20
CK_2	44140396	43228210	98.52	94.97	52.03
CK_3	39566018	38686352	98.43	94.68	51.55
IAA_1	42275074	41366510	98.51	95.00	52.46
IAA_2	47238292	46138970	98.51	94.96	52.58
IAA_3	54196830	52731944	98.51	94.96	52.27

样本Simples	原始序列数量Raw reads	清洁序列数量Valid reads	Q₂₀ （%）	Q₃₀ （%）	GC （%）
Cd_1	47397864	46440412	98.46	94.79	52.51
Cd_2	50450066	49185876	98.54	95.08	52.75
Cd_3	51288708	50076930	98.49	94.94	52.78
Cd+IAA_1	47775358	46702260	98.51	94.97	52.66
Cd+IAA_2	52741792	51346028	98.46	94.89	52.86
Cd+IAA_3	54986698	53830218	98.53	95.05	52.72
CK_1	52312058	51101542	98.48	94.87	52.20
CK_2	44140396	43228210	98.52	94.97	52.03
CK_3	39566018	38686352	98.43	94.68	51.55
IAA_1	42275074	41366510	98.51	95.00	52.46
IAA_2	47238292	46138970	98.51	94.96	52.58
IAA_3	54196830	52731944	98.51	94.96	52.27

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