1.Identification and verification of key salt-tolerant amino acid sites of banana MaNHX5.
Ya XU ; Mengxin TENG ; Fei QIAO ; Xingyu JIANG ; Yang ZHOU ; Xinguo LI
Chinese Journal of Biotechnology 2022;38(5):1946-1952
In order to improve the salt tolerance of banana NHX genes, we cloned a MaNHX5 gene from Musa acuminata L. AAA group and predicted the key salt-tolerant amino acid sites and mutant protein structure changes of MaNHX5 by using bioinformatics tools. The 276-position serine (S) of MaNHX5 protein was successfully mutated to aspartic acid (D) by site-directed mutagenesis, and the AXT3 salt-sensitive mutant yeast was used for a functional complementation test. The results showed that after the mutated MaNHX5 gene was transferred to AXT3 salt-sensitive mutant yeast, the salt tolerance of the mutant yeast was significantly improved under 200 mmol/L NaCl treatment. It is hypothesized that Ser276 of MaNHX5 protein plays an important role in the transport of Na+ across the tonoplast.
Amino Acids/metabolism*
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Gene Expression Regulation, Plant
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Musa/metabolism*
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Plant Proteins/metabolism*
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Plants, Genetically Modified
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Saccharomyces cerevisiae/metabolism*
2.Genome-wide identification of the banana GLR gene family and its expression analysis in response to low temperature and abscisic acid/methyl jasmonate.
Binbin LUO ; Hui ZHANG ; Dan LI ; Qiuzhen WU ; Wanjun GE ; Tianyuan ZHU ; Yukun CHEN ; Yuji HUANG ; Yuling LIN ; Zhongxiong LAI
Chinese Journal of Biotechnology 2023;39(7):2874-2896
Glutamate receptor-like (GLR) is an important class of Ca2+ channel proteins, playing important roles in plant growth and development as well as in response to biotic and abiotic stresses. In this paper, we performed genome-wide identification of banana GLR gene family based on banana genomic data. Moreover, we analyzed the basic physicochemical properties, gene structure, conserved motifs, promoter cis-acting elements, evolutionary relationships, and used real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) to verify the expression patterns of some GLR family members under low temperature of 4 ℃ and different hormone treatments. The results showed that there were 19 MaGLR family members in Musa acuminata, 16 MbGLR family members in Musa balbisiana and 14 MiGLR family members in Musa itinerans. Most of the members were stable proteins and had signal peptides, all of them had 3-6 transmembrane structures. Prediction of subcellular localization indicated that all of them were localized on the plasma membrane and irregularly distributed on the chromosome. Phylogenetic analysis revealed that banana GLRs could be divided into 3 subclades. The results of promoter cis-acting elements and transcription factor binding site prediction showed that there were multiple hormone- and stress-related response elements and 18 TFBS in banana GLR. RT-qPCR analysis showed that MaGLR1.1 and MaGLR3.5 responded positively to low temperature stress and were significantly expressed in abscisic acid/methyl jasmonate treatments. In conclusion, the results of this study suggest that GLR, a highly conserved family of ion channels, may play an important role in the growth and development process and stress resistance of banana.
Musa/metabolism*
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Phylogeny
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Abscisic Acid/metabolism*
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Temperature
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Stress, Physiological/genetics*
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Hormones/metabolism*
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Gene Expression Regulation, Plant
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Plant Proteins/metabolism*
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Gene Expression Profiling