Cloning of SnRK2.4 Gene and It's Response to Abiotic Stress in Kentucky Bluegrass

doi:10.7668/hbnxb.20193082

Abstract

Abstract:

Protein kinases are important factors in plant defense system,protein kinase SnRK2 is a serine/threonine protein kinase,can play an important role in the plant stress signal transduction pathway through phosphorylation.We analyzed the expression pattern of SnRK2.4 under abiotic stresses,aiming to reveal its role in the regulation of adversity.The SnRK2.4 gene of high-quality cold-season turfgrass Poa pratensis L. was cloned using RT-PCR,the SnRK2.4 gene contained an ORF of 1 092 bp encoding a 363-amino acid protein,and its molecular characteristics were analyzed by bioinformatics.In addition,the expression pattern of this gene in different tissue parts under different abiotic stresses was observed using qPCR.The results showed that the Poa pratensis L.SnRK2.4 gene belonged to the SRK/SAPK superfamily,contained typical STKc_SnRK2 domain,tyrosine kinase phosphorylation site,casein kinase Ⅱ phosphorylation site and serine/threonine-protein kinase activity site,which had the highest homology with Brachypodium distachyon.The qPCR results demonstrated that the Poa pratensis L.SnRK2.4 gene was tissue-specific,highly expressed in panicles,and had no significant differential expression in roots,stems and leaves.Moreover,the Poa pratensis L.SnRK2.4 gene could respond positively to abiotic stresses such as drought,salt,low nitrogen,low phosphorus,ABA and BR.

Key words: Kentucky bluegrass, Protein kinase, SnRK2.4, Gene cloning, Abiotic stress

JIN Yifeng, GAO Yansong, WANG Qi, WANG Mengmeng, ZHAO Di, XIONG Yi, CHEN Yang. Cloning of SnRK2.4 Gene and It's Response to Abiotic Stress in Kentucky Bluegrass[J]. Acta Agriculturae Boreali-Sinica, 2022, 37(5): 9-15. doi: 10.7668/hbnxb.20193082.

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Figures/Tables 8

Tab.1 The primers used for RT-PCR and qRT-PCR

引物名称 Primer name	序列(5'—3') Sequences(5'—3')	长度/bp Length	功能 Function
SnRK2.4-F	GACATCGGGTCGGGCAACTT	1 242	基因克隆
SnRK2.4-R	CCACTGAACTGAAATCCCCG
Q-SnRK2.4-F	TTGAAGACCCAGATGACCCC	155	qRT-PCR
Q-SnRK2.4-R	CATTGTGATTCTCCTCGCAG
Q-UBQ-F	AGAAGAAGACCTACACCAAG	217	内参基因
Q-UBQ-R	GGTTGTAAGGCGTAGGTGAG

Fig.1 Electropherogram of PCR-amplified products of SnRK2.4

Fig.2 Conserved domain analysis of SnRK2.4 in kentucky bluegrass

Fig.3 Phylogenetic tree of SnRK2.4 in kentucky bluegrass and other plants

Fig.4 Multiple sequence alignment of SnRK2.4 homologous proteins of kentucky bluegrass, Brachypodium,Hordeum vulgare and Thinopyrum elongatum Ⅰ.Protein kinase ATP binding signal domain;Ⅱ.Serine/threonine protein kinase active site;Ⅲ.Glutamate-rich region;-.cAMP and cGMP-dependent protein kinase phosphorylation site;=.N-Myristoylation site;#.Tyrosine kinase phosphorylation site;^.Casein kinase Ⅱphosphorylation site.

Fig.5 Secondary and tertiary structure of SnRK2.4 protein in kentucky bluegrass

Fig.6 Expression level analysis of SnRK2.4 in kentucky bluegrass under tissue Different lowercase letters indicate significant differences among tissue(P<0.05).

Fig.7 Expression pattern of SnRK2.4 gene in kentucky bluegrass under abiotic stress Different lowercase letters indicate significant differences during drought,salt,nitrogen,phosphorus,ABA and BR stress(P<0.05).

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