• Title/Summary/Keyword: Neurospora

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RNAi-Mediated Gene Silencing of Trcot1 Induces a Hyperbranching Phenotype in Trichoderma reesei

  • Gao, Fei;Li, Mengzhu;Liu, Weiquan;Bai, Yingguo;Tao, Tu;Wang, Yuan;Zhang, Jie;Luo, Huiying;Yao, Bin;Huang, Huoqing;Su, Xiaoyun
    • Journal of Microbiology and Biotechnology
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    • v.30 no.2
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    • pp.206-215
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    • 2020
  • Trichoderma reesei is the major filamentous fungus used to produce cellulase and there is huge interest in promoting its ability to produce higher titers of cellulase. Among the many factors affecting cellulase production in T. reesei, the mycelial phenotype is important but seldom studied. Herein, a close homolog of the Neurospora crassa COT1 kinase was discovered in T. reesei and designated TrCOT1, which is of 83.3% amino acid sequence identity. Functional disruption of Trcot1 in T. reesei by RNAi-mediated gene silencing resulted in retarded sporulation on potato dextrose agar and dwarfed colonies on minimal medium agar plates containing glucose, xylan, lactose, xylose, or glycerol as the sole carbon source. The representative mutant strain, SUS2/Trcot1i, also displayed reduced mycelia accumulation but hyperbranching in the MM glucose liquid medium, with hyphal growth unit length values decreased to 73.0 ㎛/tip compared to 239.8 ㎛/tip for the parent strain SUS2. The hyperbranching phenotype led to slightly but significantly increased cellulase secretion from 24 to 72 h in a batch culture. However, the cellulase production per unit of mycelial biomass was much more profoundly improved from 24 to 96 h.

Characterization of Mutations in AlHK1 Gene from Alternaria longipes: Implication of Limited Function of Two-Component Histidine Kinase on Conferring Dicarboximide Resistance

  • Luo, Yiyong;Yang, Jinkui;Zhu, Mingliang;Yan, Jinping;Mo, Minghe;Zhang, Keqin
    • Journal of Microbiology and Biotechnology
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    • v.18 no.1
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    • pp.15-22
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    • 2008
  • Four series (S, M, R, and W) of Alternaria longipes isolates were obtained based on consecutive selection with Dimethachlon (Dim) and ultraviolet irradiation. These isolates were then characterized according to their tolerance to Dim, sensitivity to osmotic stress, and phenotypic properties. All the selected Dim-resistant isolates showed a higher osmosensitivity than the parental strains, and the last generation was more resistant than the first generation in the M, R, and W series. In addition, the changes in the Dim resistance and osmotic sensitivity were not found to be directly correlated, and no distinct morphologic characteristics were found among the resistant and sensitive isolates, with the exception of the resistant isolate K-11. Thus, to investigate the molecular basis of the fungicide resistance, a group III two-component histidine kinase (HK) gene, AlHK1, was cloned from nineteen A. longipes isolates. AlHK1p was found to be comprised of a six 92-amino-acid repeat domain (AARD), HK domain, and response regulator domain, similar to the Os-1p from Neurospora crassa. A comparison of the nucleotide sequences of the AlHK1 gene from the Dim-sensitive and -resistant isolates revealed that all the resistant isolates contained a single-point mutation in the AARD of AlHK1p, with the exception of isolate K-11, where the AlHK1p contained a deletion of 107 amino acids. Moreover, the AlHK1p mutations in the isolates of each respective series involved the same amino acid substitution at the same site, although the resistance levels differed significantly in each series. Therefore, these findings suggested that a mutation in the AARD of AlHK1p was not the sole factor responsible for A. longipes resistance to dicarboximide fungicides.

Occurrence of an invertase producing strain of Aspergillus niger LP5 isolated from longan pollen and its application in longan syrup production to feed honey bees (Apis mellifera L.)

  • Danmek, Khanchai;Ruenwai, Rawisara;Sorachakula, Choke;Jung, Chuleui;Chuttong, Bajaree
    • Journal of Ecology and Environment
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    • v.46 no.2
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    • pp.136-143
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    • 2022
  • Background: In northern Thailand, the longan flower is the principal nectar source for honey production. Microorganisms play a critical function in the agricultural ecology. The morphological characteristics of fungal species found in longan pollen were studied. Aspergillus spp. were found to be invertase-producing strains and were employed in the longan syrup production process. The purpose of this study was to evaluate the effects of invertase-added longan syrup on the adult honey bee population numbers that were fed by this syrup for 16 weeks. Results: Different fungal species were found in longan pollen samples. Aspergillus was the main genus, with three predominant sections: Nigri, Flavi, and Terrei. Other isolated species were Trichoderma spp., Rhizopus spp., Neurospora spp., Chaetomium spp., Fusarium spp. and Penicillium spp. However, Aspergillus spp. is the only fungal species that produces the enzyme invertase. The invertase-producing strains belonging to the Aspergillus section Nigri were found to be A. niger LP5 with an optimum activity at pH 6.0 and 60℃. When A. niger LP5 invertase was used for longan syrup processing, the highest levels of glucose (3.45%) and fructose (2.08%) were found in invertase added longan syrup (C), while fresh (A) and boiled longan syrup (B) had lower contents of both sugars. The sucrose content was detected in (A) at 4.25%, while (B) and (C) were at 4.02% and 3.08%, respectively. An appropriate amount of sugar to feed and maintain the honey bee population was considered. The data showed no statistically significant differences between the two selected forms of longan syrup compared to the sugar syrup examined by the adult honey bee population. Conclusions: The main species of isolated fungi from longan pollen were Aspergillus spp. The discovery of an invertase-producing strain of A. niger LP5 has enabled its application for enzyme utilization in the invert sugar preparation process. The adult worker bee populations fed by longan syrup from both boiled and invertase-added sources showed an increasing trend. Artificial syrup made from longan fruit to feed honey bees when natural food sources are limited can be applied.

Studies on the Utilization of Sweet Potatoes for Takju Brewing (탁주양조원료(濁酒釀造原料)로서 고구마의 이용(利用)에 관(關)한 연구(硏究))

  • Kim, Chan-Jo;Choi, Woo-Young;Oh, Man-Jin
    • Applied Biological Chemistry
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    • v.15 no.3
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    • pp.213-219
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    • 1972
  • In order to utilize sweet potatoes for the material of Takju, brewing experiments with raw sweet potatoes, sweet potato chips powder and its koji were conducted; and various tests were carried out on effect of the treatments of acid, alkali, polyphenol oxidase inhibitor, oxidizing and reducing agents upon the prevention against coloring of sweet potato chips by steaming, and on peeling effect of sweet potatoes by the alkali and heat treatments. The results obtained were as follows. 1) In the case of brewing with raw sweet potatose, each plot showed low acid and ethanol content, and its finished Takju had an undersirable color and odor. The plots which were mashed after peeling showed higher ethanol contents than the plots mashed without peeling. 2) In the case of brewing with sweet potato chips powder, each plot contained considerably more amount of ethanol than the plots brewed with raw sweet potatoes, white it contained less amount of acid. The ethanol contents of the plots using wheat bran koji were $10.5{\sim}11.4$ per cent 4 days after mashing, and were higher than those of the plots using malts powder. Their finished Takju was inferior in quality because of the lack of acid and being darkened gradually in process of time. 3) The kojies which were made of sweet potato chips powder with Neurospora sitophila or Aspergillus oryzae had good appearance, but the Takju mashes brewed with these contained remarkably less amount of ethanol. 4) Effect of the treatments of acid, alkali, polyphenol oxidase inhibitor and organic solvents such as ether and ethanol upon the prevention against coloring of sweet potato chips was not recognized. Alum and burnt alum were effective a little on the decolorization, and among the oxidizing and reducing agents tested, potassium permanganate was most effective. 5) Darkening of sweet potato chips powder in course of heating after mixing with water was not affected by pectin and amino acids, but by tannin. 6) Sweet potatoes were not peeled easily by friction after soaking in the boiling solution of 3 per cent alkali for 6 minutes and peeled in boiling water for 12 minutes. From the viewpoint of the results above mentioned, it seems to be necessary to study further on the isolation of microorganisms which are able to decompose the coloring substances and yeasts which are adequate for the fermentation of sweet potatoes in order to utilize sweet potatoes for Takju brewing, because brewing with raw sweet potatoes, sweet potato chips powder and its koji was unsuccessful, and effect of the various treatments on the decolorization of sweet potatoes was not recognized.

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