28 research outputs found
The yeast-one-hybrid assay identifies LHCA2 and HSPRO2 as double-SORLIP1 element binding proteins in Arabidopsis thaliana
No full textEarly light induced proteins (ELIPs) are widely distributed in the plant kingdom. Members of the extended light harvesting complex (LHC) superfamily, ELIPs are expressed in the nucleus and the ELIP protein is transiently localized to the thylakoid membranes. Significant increase in expression of ELIPs has been reported in response to stresses such as high light, high and low temperature, exposure to UV and salinity. ELIP expression also increases at transitional stages of chloroplast development such as deetiolation, conversion to chromoplast, and senescence. In search of cis-regulatory regions, the A. thaliana ELIP1 gene promoter has been investigated in our lab. A double-SORLIP1 element was identified as a critical cis-regulatory region common in the promoters of A. thaliana ELIP1 (At3g22840) and ELIP2 (At4g14690). Point mutations in the double-SORLIP1 element led to significant decline in expression. Due to the importance of the double-SORLIP1 element, a yeast-one-hybrid assay was set up to find the specific DNA-binding proteins that bind to this region. Light harvesting complex II (LHCA2) and the ortholog of sugar beet HS1 PRO-1 2, heat-shock-like protein 2 (HSPRO2), showed a high specificity in binding to the double-SORLIP1 element. Investigation of lhca2 and hspro2 Arabidopsis mutants did not show any significant difference in high light induced expression of ELIP1 or ELIP2 as compared to wild type. However, the high frequency of LHCA2 clones selected by yeast-one-hybrid assay and the high specificity in binding to the double-SORLIP1 element cannot be ignored. After reviewing the literature, I hypothesized that LHCA2 may be a new retrograde signal that regulates expression of ELIP genes. However, more experimental evidence is needed to support this proposed function. The potential regulatory role of HSPRO2 is also discussed
Evaluation of Structural Features of Membrane Acting Antifungal Peptides by Artificial Neural Network
No full textA double SORLIP1 element is required for high light induction of ELIP genes in Arabidopsis thaliana
No full textA double SORLIP1 element is required for high light induction of ELIP genes in Arabidopsis thaliana
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Unraveling the regulation of sugar beet pulp utilization in the industrially relevant fungus Aspergillus niger
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Multilevel analysis between Physcomitrium patens and Mortierellaceae endophytes explores potential long‐standing interaction among land plants and fungi
No full textThe model moss species Physcomitrium patens has long been used for studying divergence of land plants spanning from bryophytes to angiosperms. In addition to its phylogenetic relationships, the limited number of differential tissues, and comparable morphology to the earliest embryophytes provide a system to represent basic plant architecture. Based on plant-fungal interactions today, it is hypothesized these kingdoms have a long-standing relationship, predating plant terrestrialization. Mortierellaceae have origins diverging from other land fungi paralleling bryophyte divergence, are related to arbuscular mycorrhizal fungi but are free-living, observed to interact with plants, and can be found in moss microbiomes globally. Due to their parallel origins, we assess here how two Mortierellaceae species, Linnemannia elongata and Benniella erionia, interact with P. patens in coculture. We also assess how Mollicute-related or Burkholderia-related endobacterial symbionts (MRE or BRE) of these fungi impact plant response. Coculture interactions are investigated through high-throughput phenomics, microscopy, RNA-sequencing, differential expression profiling, gene ontology enrichment, and comparisons among 99 other P. patens transcriptomic studies. Here we present new high-throughput approaches for measuring P. patens growth, identify novel expression of over 800 genes that are not expressed on traditional agar media, identify subtle interactions between P. patens and Mortierellaceae, and observe changes to plant-fungal interactions dependent on whether MRE or BRE are present. Our study provides insights into how plants and fungal partners may have interacted based on their communications observed today as well as identifying L. elongata and B. erionia as modern fungal endophytes with P. patens
The transcriptional activator ClrB is crucial for the degradation of soybean hulls and guar gum in Aspergillus niger
No full textLow-cost plant substrates, such as soybean hulls, are used for various industrial applications. Filamentous fungi are important producers of Carbohydrate Active enZymes (CAZymes) required for the degradation of these plant biomass substrates. CAZyme production is tightly regulated by several transcriptional activators and repressors. One such transcriptional activator is CLR-2/ClrB/ManR, which has been identified as a regulator of cellulase and mannanase production in several fungi. However, the regulatory network governing the expression of cellulase and mannanase encoding genes has been reported to differ between fungal species. Previous studies showed that Aspergillus niger ClrB is involved in the regulation of (hemi-)cellulose degradation, although its regulon has not yet been identified. To reveal its regulon, we cultivated an A. niger ΔclrB mutant and control strain on guar gum (a galactomannan-rich substrate) and soybean hulls (containing galactomannan, xylan, xyloglucan, pectin and cellulose) to identify the genes that are regulated by ClrB. Gene expression data and growth profiling showed that ClrB is indispensable for growth on cellulose and galactomannan and highly contributes to growth on xyloglucan in this fungus. Therefore, we show that A. niger ClrB is crucial for the utilization of guar gum and the agricultural substrate, soybean hulls. Moreover, we show that mannobiose is most likely the physiological inducer of ClrB in A. niger and not cellobiose, which is considered to be the inducer of N. crassa CLR-2 and A. nidulans ClrB
Recommended from our members
The transcriptional activator ClrB is crucial for the degradation of soybean hulls and guar gum in Aspergillus niger
No full textLow-cost plant substrates, such as soybean hulls, are used for various industrial applications. Filamentous fungi are important producers of Carbohydrate Active enZymes (CAZymes) required for the degradation of these plant biomass substrates. CAZyme production is tightly regulated by several transcriptional activators and repressors. One such transcriptional activator is CLR-2/ClrB/ManR, which has been identified as a regulator of cellulase and mannanase production in several fungi. However, the regulatory network governing the expression of cellulase and mannanase encoding genes has been reported to differ between fungal species. Previous studies showed that Aspergillus niger ClrB is involved in the regulation of (hemi-)cellulose degradation, although its regulon has not yet been identified. To reveal its regulon, we cultivated an A. niger ΔclrB mutant and control strain on guar gum (a galactomannan-rich substrate) and soybean hulls (containing galactomannan, xylan, xyloglucan, pectin and cellulose) to identify the genes that are regulated by ClrB. Gene expression data and growth profiling showed that ClrB is indispensable for growth on cellulose and galactomannan and highly contributes to growth on xyloglucan in this fungus. Therefore, we show that A. niger ClrB is crucial for the utilization of guar gum and the agricultural substrate, soybean hulls. Moreover, we show that mannobiose is most likely the physiological inducer of ClrB in A. niger and not cellobiose, which is considered to be the inducer of N. crassa CLR-2 and A. nidulans ClrB
Unraveling the regulation of sugar beet pulp utilization in the industrially relevant fungus Aspergillus niger
No full textEfficient utilization of agro-industrial waste, such as sugar beet pulp, is crucial for the bio-based economy. The fungus Aspergillus niger possesses a wide array of enzymes that degrade complex plant biomass substrates, and several regulators have been reported to play a role in their production. The role of the regulators GaaR, AraR, and RhaR in sugar beet pectin degradation has previously been reported. However, genetic regulation of the degradation of sugar beet pulp has not been assessed in detail. In this study, we generated a set of single and combinatorial deletion mutants targeting the pectinolytic regulators GaaR, AraR, RhaR, and GalX as well as the (hemi-)cellulolytic regulators XlnR and ClrB to address their relative contribution to the utilization of sugar beet pulp. We show that A. niger has a flexible regulatory network, adapting to the utilization of (hemi-)cellulose at early timepoints when pectin degradation is impaired
