Isolation and Functional Analysis of Garlic Calmodulin Genes AsCML15 and AsCML42 under Osmotic Stress

doi:10.7668/hbnxb.20194748

Abstract

Abstract:

Calmodulin-like proteins(CMLs),one of the Ca²⁺ receptors in plants,are involved in the process of plant growth and development,as well as adaptation to environmental changes.To understand the sequence characteristics of garlic CMLs and their responses to osmotic stress,AsCML15 and AsCML42 genes from garlic variety Cangshan siliuban were cloned,and their expression patterns under drought and salt stress conditions were determined.The results showed that the open reading frame of AsCML15 and AsCML42 genes were 498 and 543 bp in length,respectively,encoding 165 and 180 amino acid residues.AsCML15 and AsCML42 harbored four and three EF-hand domains,respectively.AsCML42 was closer to Arabidopsis AtCML42 and AtCML43 in evolutionary relationship,whereas AsCML15 was more closely related to Arabidopsis AtCML15 and AtCML16.Real-time Quantitative PCR technology showed that AsCML15 and AsCML42 were expressed in bulbs,leaves,and roots,and these two genes can be induced by 200 mmol/L NaCl and 15% PEG6000.The AsCML15 and AsCML42 genes may be involved in the process of garlic resisting salt and drought stress,and their biological functions can be further identified.

Key words: Garlic, Calmodulin-like protein, Gene cloning, Expression analysis, Osmotic stress

WANG Guanglong, XU Wujun, CHEN Yangyang, HU Zhenzhu, SUN Min, XIONG Aisheng. Isolation and Functional Analysis of Garlic Calmodulin Genes AsCML15 and AsCML42 under Osmotic Stress[J]. Acta Agriculturae Boreali-Sinica, 2024, 39(4): 80-86. doi: 10.7668/hbnxb.20194748.

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Figures/Tables 8

Tab.1 Primer sequences applied for AsCML15 and AsCML42 cloning and Quantitative Real-time PCR

用途 Function	基因名称 Gene name	上游引物(5'—3') Forward primer(5'—3')	下游引物(5'—3') Reverse primer(5'—3')
克隆	AsCML42	ATGTCTCCAGCTCCTCCATCAAAGA	CTAAGATTGAGCAACAACACTCCTC
Gene cloning	AsCML15	ATGGTTGGAAAATGCAAGCTACGGA	TCAATTAGCGACGGCATGAACGCCC
荧光定量PCR	AsCML42	GAGCACATCCTCCACCTCATTCC	CATCCACACTTCCGATCATCACCAT
Quantitative	AsCML15	GATCACCTGAACCAACTCCGAGAG	AGCTCCGCCGCCGATATGTA
Real-time PCR	AsSAND	GCGTCAACGAATGTTCCAATTACCA	TCTCTTCAGTCTCAACTTCATCAGCAT

Fig.1 RT-PCR amplification pattern of AsCML15 and AsCML42 genes from garlic M.DNA Marker;1.AsCML42 gene;2.AsCML15 gene.

Fig.2 Nucleotide acid and predicted amino acid sequences of AsCML15 and AsCML42 genes from garlic

Fig.3 Prediction of hydrophilicity of AsCML15 and AsCML42 from garlic

Fig.4 Alignment of amino acids of CML15 and CML42 from different species ^*.Ca²⁺ binding sites.

Fig.5 Phylogenetic tree of AsCML15 and AsCML42 from garlic and CML from Arabidopsis

Fig.6 Expression patterns of AsCML15 and AsCML42 genes in garlic under simulated drought stress Different lowercase letters within the same tissues indicate significant differences at the 0.05 level.The same as Fig.7.

Fig.7 Expression patterns of AsCML15 and AsCML42 genes in garlic under salt stress

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