Accumulation of copper in Trichoderma reesei transformants, constructed with the modified Agrobacterium tumefaciens-mediated transformation technique

Fu, Kehe; Liu, Lixing; Fan, Lili; Tong, Le; Chen, Jie

doi:10.1007/s10529-010-0365-y

Cited by 10 publications

(2 citation statements)

References 11 publications

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“…High frequencies of multicopy insertion and ectopic integration have been observed in the application of these filamentous fungal transformation systems [ 25 , 26 ]. Compared with PEG/CaCl 2 protoplast, electroporation and biolistic transformation systems, A. tumefaciens -mediated transformation alleviates variable multicopy insertion and ectopic integration with its high efficiency (>90%) of single-copy integration in T. reesei [ 27 ]. In this study, we introduced the Cas9-encoding gene into the genome of T. reesei using random A. tumefaciens -mediated transformation.…”

Section: Discussionmentioning

confidence: 99%

Efficient genome editing in filamentous fungus Trichoderma reesei using the CRISPR/Cas9 system

et al. 2015

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Filamentous fungi have wide applications in biotechnology. The CRISPR/Cas9 system is a powerful genome-editing method that facilitates genetic alterations of genomes in a variety of organisms. However, a genome-editing approach has not been reported in filamentous fungi. Here, we demonstrated the establishment of a CRISPR/Cas9 system in the filamentous fungus Trichoderma reesei by specific codon optimization and in vitro RNA transcription. It was shown that the CRISPR/Cas9 system was controllable and conditional through inducible Cas9 expression. This system generated site-specific mutations in target genes through efficient homologous recombination, even using short homology arms. This system also provided an applicable and promising approach to targeting multiple genes simultaneously. Our results illustrate that the CRISPR/Cas9 system is a powerful genome-manipulating tool for T. reesei and most likely for other filamentous fungal species, which may accelerate studies on functional genomics and strain improvement in these filamentous fungi.

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Section: Discussionmentioning

confidence: 99%

Efficient genome editing in filamentous fungus Trichoderma reesei using the CRISPR/Cas9 system

et al. 2015

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“…Copper stress causes the accumulation of large amounts of reactive oxygen species in cells [ 11 , 12 ]. Generally, it is believed that cells activate and increase the levels of antioxidant enzymes such as CAT, POD, and SOD and reduce that of glutathione (GSH) to alleviate oxidative stress [ 13 ].…”

Section: Discussionmentioning

confidence: 99%

Functions of the C2H2 Transcription Factor Gene thmea1 inTrichoderma harzianumunder Copper Stress Based on Transcriptome Analysis

Mei

Wang

Jiang

et al. 2018

BioMed Research International

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Trichoderma spp. are important biocontrol filamentous fungi and have tremendous potential in soil bioremediation. In our previous studies, a C2H2 type transcription factor coding gene (thmea1) was cloned from a biocontrol agent T. harzianum Th-33; the encoded sequence of thmea1 contained 3 conserved C2H2 domains with Swi5 and Ace2 in Saccharomyces cerevisiae. The thmea1 knockout mutant Δthmea1 showed 12.9% higher copper tolerance than the wild-type Th33. To elucidate the function of thmea1 and its relationship with copper stress response, we conducted transcriptome sequencing and analysis of wild-type Th33 and Δthmea1 under 0.8 mM copper stress. A total of 1061 differentially expressed genes (DEGs) were identified between the two strains, all DEGs were assigned to KEGG pathway database, 383 DEGs were annotated in 191 individual pathways, and the categories of ribosomal protein synthesis and amino acid metabolism were the most highly enriched ones. Analysis of related DEGs showed that the expression levels of intracellular glutathione detoxification enzyme, heat shock proteins, and ribosomal proteins in Δthmea1 were higher than that of the wild-type Th33, and the expression of metallothionein (MT) gene did not change. In addition, the expression levels of genes coding for proteins associated with the Ccc2p-mediated copper chaperone Atx1p transport of copper ions into the Golgi secretory pathway increased, as well as the copper amine oxidase (CuAO). These findings suggest that Thmea1 is a negative regulated factor of copper tolerance ability in T. harzianum. It does not show metallothionein expression activator activities as that of Ace2 in S. cerevisiae. We hypothesize that after T. harzianum has lost its thmea1 gene, the ability of cells to scavenge reactive oxygen species, mainly through the glutathione antioxidant system, is enhanced, whereas protein synthesis and repair and copper secretion increase under copper stress, which increases the ability of the mutant strain to tolerate copper stress.

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Bioremoval of lead and iron from sewage water by mangrove-derived Hypocrea lixii

Saravanakumar

Kathiresan

2014

Int. J. Environ. Sci. Technol.

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This work focused on the bioremoval of heavy metals by mangrove-derived Trichoderma biomass in the artificial sewage water. Of the 12 isolates tested, Hypocrea lixii TSK8 (JQ809340) was found to be a potent strain in removal of lead and iron. The bioremoval was enhanced through adsorption kinetics and process optimization using statistical model of the response surface methodology. The magnitude of Langmuir constant was 0.64 L mg -1 and 0.42 L mg -1 for lead and iron, respectively. Adsorption capacity was determined as 49.2 mg g -1 and 51.3 mg g -1for lead and iron, respectively. The optimized conditions for maximum removal of lead and iron were standardized. The biosorption of metals was also confirmed by scanning electron microscopic and X-ray energy-dispersive spectrometer analyses. The results revealed that dried biomass of the H. lixii TSK8 was a potent biosorbent for efficient bioremoval of lead and iron.

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Accumulation of copper in Trichoderma reesei transformants, constructed with the modified Agrobacterium tumefaciens-mediated transformation technique

Cited by 10 publications

References 11 publications

Efficient genome editing in filamentous fungus Trichoderma reesei using the CRISPR/Cas9 system

Efficient genome editing in filamentous fungus Trichoderma reesei using the CRISPR/Cas9 system

Functions of the C2H2 Transcription Factor Gene thmea1 inTrichoderma harzianumunder Copper Stress Based on Transcriptome Analysis

Bioremoval of lead and iron from sewage water by mangrove-derived Hypocrea lixii

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