Cloning, Heterologous Expression, and Functional Characterization of a Chitinase Gene, Lbchi32, from Limonium bicolor

Liu, Zhi Hua; Yang, Chuan Ping; Qi, Xiao Tian; Xiu, Li; Wang, Yu Cheng

doi:10.1007/s10528-010-9348-x

Cited by 14 publications

(6 citation statements)

References 25 publications

(25 reference statements)

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“…Five out of the 15 enriched GO terms in Triticeae can be related to resistance to fungal pathogens via three different mechanisms. Chitinases are enzymes, which are known to be involved in plants' fungal resistance and have been extensively studied in wheat species [28,29]. The ability of these enzymes to degrade chitin, a primary component of fungal cell walls, can lead to the lysis of fungal cells and therefore provide resistance against them.…”

Section: Resistance To Fungi In Wheatmentioning

confidence: 99%

The modular nature of protein evolution: domain rearrangement rates across eukaryotic life

et al. 2020

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Background: Modularity is important for evolutionary innovation. The recombination of existing units to form larger complexes with new functionalities spares the need to create novel elements from scratch. In proteins, this principle can be observed at the level of protein domains, functional subunits which are regularly rearranged to acquire new functions. Results: In this study we analyse the mechanisms leading to new domain arrangements in five major eukaryotic clades (vertebrates, insects, fungi, monocots and eudicots) at unprecedented depth and breadth. This allows, for the first time, to directly compare rates of rearrangements between different clades and identify both lineage specific and general patterns of evolution in the context of domain rearrangements. We analyse arrangement changes along phylogenetic trees by reconstructing ancestral domain content in combination with feasible single step events, such as fusion or fission. Using this approach we explain up to 70% of all rearrangements by tracing them back to their precursors. We find that rates in general and the ratio between these rates for a given clade in particular, are highly consistent across all clades. In agreement with previous studies, fusions are the most frequent event leading to new domain arrangements. A lineage specific pattern in fungi reveals exceptionally high loss rates compared to other clades, supporting recent studies highlighting the importance of loss for evolutionary innovation. Furthermore, our methodology allows us to link domain emergences at specific nodes in the phylogenetic tree to important functional developments, such as the origin of hair in mammals. Conclusions: Our results demonstrate that domain rearrangements are based on a canonical set of mutational events with rates which lie within a relatively narrow and consistent range. In addition, gained knowledge about these rates provides a basis for advanced domain-based methodologies for phylogenetics and homology analysis which complement current sequence-based methods.

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Section: Resistance To Fungi In Wheatmentioning

confidence: 99%

The modular nature of protein evolution: domain rearrangement rates across eukaryotic life

et al. 2020

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“…Though many mutants in salt secretion have been obtained using gamma ray irradiation ( Yuan et al, 2013 ), no recretohalophyte where specific genes have been silenced has been developed. Until now, the experiments suggesting candidate genes have only been reported in heterologous expression, such as Lbchi32 of L. bicolor verified in pathogenic fungi ( Liu et al, 2010 ). So the mutants in specific and candidate genes should be investigated in the future in order to accurately study molecular mechanism of salt secretion.…”

Section: A Possible Pathway For Secreting Salt From the Salt Glandmentioning

confidence: 99%

Progress in Studying Salt Secretion from the Salt Glands in Recretohalophytes: How Do Plants Secrete Salt?

2016

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To survive in a saline environment, halophytes have evolved many strategies to resist salt stress. The salt glands of recretohalophytes are exceptional features for directly secreting salt out of a plant. Knowledge of the pathway(s) of salt secretion in relation to the function of salt glands may help us to change the salt-tolerance of crops and to cultivate the extensive saline lands that are available. Recently, ultrastructural studies of salt glands and the mechanism of salt secretion, particularly the candidate genes involved in salt secretion, have been illustrated in detail. In this review, we summarize current researches on salt gland structure, salt secretion mechanism and candidate genes involved, and provide an overview of the salt secretion pathway and the asymmetric ion transport of the salt gland. A new model recretohalophyte is also proposed.

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“…The activities of four different recombinant chitinases, inrCHI31, exrCHI31, inrCHI32, and exrCHI32, towards different substrates were measured using Schales method [15]. While four recombinant chitinases demonstrated activity as a response to all test substrates, analysis of variance showed that their decomposing activities towards different substrates were significantly different ( P < 0.05) (Figure 5).…”

Section: Resultsmentioning

confidence: 99%

“…The optimal temperature and pH for chitinase activity, thermal stability, and the effects of ions on enzyme activity, and the rate of decomposition of fungal cell walls of recombinant chitinases inrCHI31, exrCHI31, inrCHI32, and exrCHI32 were investigated according to the technique described by Liu et al [14, 15]. …”

Section: Methodsmentioning

confidence: 99%

Chitinase Genes LbCHI31 and LbCHI32 from Limonium bicolor Were Successfully Expressed in Escherichia coli and Exhibit Recombinant Chitinase Activities

Liu

Huang

Zhang

et al. 2013

The Scientific World Journal

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The two chitinase genes, LbCHI31 and LbCHI32 from Limonium bicolor, were, respectively, expressed in Escherichia coli BL21 strain. The intracellular recombinant chitinases, inrCHI31 and inrCHI32, and the extracellular exrCHI31 and exrCHI32 could be produced into E. coli. The exrCHI31 and exrCHI32 can be secreted into extracellular medium. The optimal reaction condition for inrCHI31 was 5 mmol/L of Mn2+ at 40°C and pH 5.0 with an activity of 0.772 U using Alternaria alternata cell wall as substrate. The optimal condition of inrCHI32 was 5 mmol/L of Ba2+ at 45°C and pH 5.0 with an activity of 0.792 U using Valsa sordida cell wall as substrate. The optimal reaction condition of exrCHI31 was 5 mmol/L of Zn2+ at 40°C and pH 5.0, and the activity was 0.921 U using the A. alternata cell wall as substrate. Simultaneously, the optimal condition of exrCHI32 was 5 mmol/L of K+ at 45°C and pH 5.0, with V. sordida cell wall as the substrate, and the activity was 0.897 U. Furthermore, the activities of extracellular recombinant enzymes on fungal cell walls and compounds were generally higher than those of the intracellular recombinant enzymes. Recombinant exrCHI31 and exrCHI32 have better hydrolytic ability on cell walls of different fungi than synthetic chitins and obviously showed activity against A. alternata.

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Cloning, Heterologous Expression, and Functional Characterization of a Chitinase Gene, Lbchi32, from Limonium bicolor

Cited by 14 publications

References 25 publications

The modular nature of protein evolution: domain rearrangement rates across eukaryotic life

The modular nature of protein evolution: domain rearrangement rates across eukaryotic life

Progress in Studying Salt Secretion from the Salt Glands in Recretohalophytes: How Do Plants Secrete Salt?

Chitinase Genes LbCHI31 and LbCHI32 from Limonium bicolor Were Successfully Expressed in Escherichia coli and Exhibit Recombinant Chitinase Activities

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