CRISPR-Cas9-mediated seb1 disruption in Talaromyces pinophilus EMU for its enhanced cellulase production

Manglekar, Rupali Rahul; Geng, Anli

doi:10.1016/j.enzmictec.2020.109646

Cited by 22 publications

(5 citation statements)

References 33 publications

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“…The disruption of the seb1 gene in strain EMU resulted in morphology defects and more hyphal branching in seb1 mutant strains. The transcription levels of rhoA genes were reduced [ 51 ]. The down-regulated expression of RhoA was found in both PeGNAI -OE and PeGNAI -RNAi strains ( Figure 7 A), which could be responsible for the more luxuriant aerial mycelium of the transformed strains ( Figure 1 ).…”

Section: Discussionmentioning

confidence: 99%

Transcriptome Analysis Reveals the Function of a G-Protein α Subunit Gene in the Growth and Development of Pleurotus eryngii

Cao

Sun

et al. 2023

JoF

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Pleurotus eryngii is a commercially important edible fungus with high nutritional and economic value. However, few functional studies have examined key genes affecting the growth and development of P. eryngii. In this study, transformed strains, including over-expression (PeGNAI-OE) and RNA interference (PeGNAI-RNAi) lines, were constructed to elucidate the role of GNAI in P. eryngii growth. GNAI expression was found to affect the mycelial growth and the number of clamp connections. Moreover, the transformed strains were shown to have higher endogenous cAMP levels, thus affecting amylase and laccase activity. Fruiting experiments showed that GNAI expression revealed the formation of P. eryngii primordia and the number of buttons, while transcription analysis identified GNAI gene involvement in the growth and development of P. eryngii. Seven downstream genes regulated by GNAI were differentially expressed in PeGNAI-OE and PeGNAI-RNAi compared to wild type (WT). These genes may be related to mycelial growth and enzyme activity. They were involved in the MAPK signaling pathway, inositol phosphate metabolism, ascorbate, aldarate metabolism, and starch and sucrose metabolism. In summary, GNAI performs different physiological functions in regulating the growth and development of P. eryngii. Importantly, the molecular mechanisms of GNAI regulatory function are relatively complex and need further study.

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Section: Discussionmentioning

confidence: 99%

Transcriptome Analysis Reveals the Function of a G-Protein α Subunit Gene in the Growth and Development of Pleurotus eryngii

Cao

Sun

et al. 2023

JoF

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“…Therefore, the effects of CSDI mode on soil health and crop quality need to be evaluated further. In addition, Mortierella and Fusarium have a high cellulase secretion capacity to effectively degrade plant residues (Ellegaard‐Jensen et al, 2013; Manglekar & Geng, 2020), and thus performing an important role in SOC accumulation (Beccari et al, 2011; Clemmensen et al, 2015).…”

Section: Discussionmentioning

confidence: 99%

Deep incorporation of corn straw benefits soil organic carbon and microbial community composition in a black soil of Northeast China

Zhu

Lin

Guan

et al. 2022

Soil Use and Management

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‘Deep incorporation of corn straw’ (CSDI) is to concentrate the burial of corn straw into the subsurface soil layer (20–40 cm) and to break the plough pan, thereby creating a loosened plough layer (0–20 cm) and a fertile subsurface soil layer. However, its impacts on soil organic carbon (SOC) and the microbial community remain poorly understood. A field experiment was conducted to investigate the effects of 1‐year CSDI (CD1), 3‐year CSDI (CD3) and 5‐year CSDI (CD5) on soil aggregates and aggregate‐associated SOC, as well as bacterial and fungal community characteristics (examined by the high‐throughput gene sequencing method). The results demonstrated that SOC and soil fungal diversity were decreased by CD1, but increased by CD3 and CD5. Compared with the control, CD5 promoted 2–0.25 mm soil macroaggregation, significantly increased SOC by 8.94% and aggregate‐associated SOC by 5.96%–8.84%, consequently improving the physical protection of SOC by soil aggregates. CD3 and CD5 enhanced the richness and diversity of soil bacteria and fungi, and altered community composition. For soil bacteria, the relative abundance of Acidobacteria and Chloroflexi was increased, while that of Firmicutes, Gemmatimonadetes, Sphingomonas and Bacillus was decreased. For soil fungi, the relative abundance of Ascomycota, Zygomycota, Mortierella and Fusarium was greatly improved, but that of Basidiomycota was reduced. These obvious variations in microbial community structure were beneficial to straw degradation and SOC accumulation. Overall, the optimization of microbial community with CSDI plays a positive role in promoting soil organic matter, nutrient cycling and carbon sequestration, and thus improving soil fertility.

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“…In the past few years, CRISPR/Cas has emerged as the preferred technique for genome editing due to their innate ability to generate indels in the target genes mediated by RNA‐guided endonuclease (Cong et al, 2013; Doudna & Charpentier, 2014; Jinek et al, 2012). Since the advent of CRISPR/Cas9 technology, it has been used successfully for generating the mutant for enhanced production of cellulase in Myceliophthora thermophila (Li et al, 2020; Liu et al, 2019) and Talaromyces pinophilus (Manglekar & Geng, 2020) as well as amylase in Myceliopthora thermophila (Li et al, 2020). However, CRISPR/Cas9 system has several inherent limitations including protospacer adjacent motif (PAM) dependency, generation of blunt ends and high propensity of the off‐target mutations (Zetsche et al, 2015).…”

Section: New Precision Approaches: Strain Improvement and Enhanced En...mentioning

confidence: 99%

Plant-associated endophytic fungi as potential bio-factories for extracellular enzymes: Progress, Challenges and Strain improvement with precision approaches

Raghav

Jyoti

Siddiqui

et al. 2022

Journal of Applied Microbiology

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There is an intricate network of relations between endophytic fungi and their hosts that affects the production of various bioactive compounds. Plant-associated endophytic fungi contain industrially important enzymes and have the potential to fulfil their rapid demand in the international market to boost business in technology.Being safe and metabolically active, they have replaced the usage of toxic and harmful chemicals and hold a credible application in biotransformation, bioremediation and industrial processes. Despite these, there are limited reports on fungal endophytes that can directly cater to the demand and supply of industrially stable enzymes. The underlying reasons include low endogenous production and secretion of enzymes from fungal endophytes which have raised concern for widely accepted applications. Hence, it is imperative to augment the biosynthetic and secretory potential of fungal endophytes. Modern state-of-the-art biotechnological technologies aiming at strain improvement using cell factory engineering as well as precise gene editing like Clustered Regularly Interspaced Palindromic Repeats (CRISPR) and its Associated proteins (Cas) systems which can provide a boost in fungal endophyte enzyme production. Additionally, it is vital to characterize optimum conditions to grow one strain with multiple enzymes (OSME). The present review encompasses various plants-derived endophytic fungal enzymes and their applications in various sectors. Furthermore, we postulate the feasibility of new precision approaches with an aim for strain improvement and enhanced enzyme production.

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CRISPR-Cas9-mediated seb1 disruption in Talaromyces pinophilus EMU for its enhanced cellulase production

Cited by 22 publications

References 33 publications

Transcriptome Analysis Reveals the Function of a G-Protein α Subunit Gene in the Growth and Development of Pleurotus eryngii

Transcriptome Analysis Reveals the Function of a G-Protein α Subunit Gene in the Growth and Development of Pleurotus eryngii

Deep incorporation of corn straw benefits soil organic carbon and microbial community composition in a black soil of Northeast China

Plant-associated endophytic fungi as potential bio-factories for extracellular enzymes: Progress, Challenges and Strain improvement with precision approaches

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