Universal primers for amplifying the complete coding sequence of cytoplasmic heat shock protein 90 (HSP90) in Lepidoptera

Xu, Peng; Li, Tong; Xiao, Jin; Murphy, Robert W.; Wei, Huang

doi:10.14411/eje.2011.020

Cited by 4 publications

(2 citation statements)

References 29 publications

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“…Five loci were used as indicators of stress, being associated with G-protein coupled receptor activity and stress response (contig 20595), haemocyte migration and cell proliferation (contigs 704 and 233), anti-apoptosis activity (6GM contig 5976), and receptor activity for inflammatory mediators (2GM contig 20004). These genes were selected following pre-screening, and primers were designed from published G. mellonella sequences (NCBI), from a G. mellonella EST library [26] and from lepidopteran Hsp-90 primers described by Xu et al [28]. Full details are in the electronic supplementary material, § §S1.11-S1.12 and table S1.…”

Section: (I) Qrt-pcr Analysis Of Putative Stress Management Genesmentioning

confidence: 99%

More than a colour change: insect melanism, disease resistance and fecundity

et al. 2013

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A 'dark morph' melanic strain of the greater wax moth, Galleria mellonella, was studied for its atypical, heightened resistance to infection with the entomopathogenic fungus, Beauveria bassiana. We show that these insects exhibit multiple intraspecific immunity and physiological traits that distinguish them from a non-melanic, fungus-susceptible morph. The melanic and non-melanic morphs were geographical variants that had evolved different, independent defence strategies. Melanic morphs exhibit a thickened cuticle, higher basal expression of immunity-and stress-management-related genes, higher numbers of circulating haemocytes, upregulated cuticle phenoloxidase (PO) activity concomitant with conidial invasion, and an enhanced capacity to encapsulate fungal particles. These insects prioritize specific augmentations to those frontline defences that are most likely to encounter invading pathogens or to sustain damage. Other immune responses that target late-stage infection, such as haemolymph lysozyme and PO activities, do not contribute to fungal tolerance. The net effect is increased larval survival times, retarded cuticular fungal penetration and a lower propensity to develop haemolymph infections when challenged naturally (topically) and by injection. In the absence of fungal infection, however, the heavy defence investments made by melanic insects result in a lower biomass, decreased longevity and lower fecundity in comparison with their non-melanic counterparts. Although melanism is clearly correlated with increased fungal resistance, the costly mechanisms enabling this protective trait constitute more than just a colour change.

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Section: (I) Qrt-pcr Analysis Of Putative Stress Management Genesmentioning

confidence: 99%

More than a colour change: insect melanism, disease resistance and fecundity

et al. 2013

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“…The lack of positive result in the detection of the encoding genes comes from the unknown mechanism replaces the encoding genes (SÖderholm and Jaakkola 2013) or the absence of the encoding genes (Nakazawa and Honda 2015) or the unspecific attachment of the primers (Kumar et al 2017). In this case, designing the specific primer is needed to obtain the appropriate result (Xu et al 2011).…”

Section: Discussionmentioning

confidence: 99%

First report of seaweed-associated yeast from Indonesia: Species composition and screening of their polysaccharides-degrading enzymes

et al. 2022

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Abstract. Sibero MT, Frederick EH, Sabdono A, Wijayanti DP, Pringgenies D, Radjasa OK, Zilda DS, Murwani R. 2022. First report of seaweed-associated yeast from Indonesia: Species composition and screening of their polysaccharides-degrading enzymes. Biodiversitas 23: 1408-1419. Yeast has been widely utilized in various industries due to its enzyme properties. Therefore, plenty of studies focus on exploring yeast biodiversity from diverse sources. However, there are limited reports regarding marine yeast biodiversity and its potential from Indonesia. This study aimed to isolate and identify the seaweed-associated yeast from Jepara in Central Java and Sepanjang Beach in Gunung Kidul, Yogyakarta, then examined their potential to produce extracellular polysaccharides-degrading enzymes (EPEs). Marine yeasts were isolated using standard marine agar (STD) and potato dextrose agar (PDA). All isolates were characterized by their salinity tolerance, morphology, and species confirmation using DNA barcoding. Three EPEs consisting of agarase, alginate-lyase, and carrageenase were screened using polysaccharides-enriched agar media. A specific agarase encoding gene was detected with specific primers. In total, 21 seaweed-associated yeast were successfully isolated from 6 seaweeds and noted as facultative marine yeast. The DNA barcoding study discovered that these yeasts belonged to 5 genera, consisted of Aureobasidium, Candida, Debaryomyces, Hortaea, and Rhodotorula. Moreover, Candida was noted as the most abundant genus (71.42%). It was noted that Hortaea werneckii MTJ.11 and MTJ.13 were positive for all enzymes; Aureobasidium melanogenum MTGK.31 gave a positive result for agarase and carrageenase; while [Candida] zeylanoides MTGK.23 only exhibited alginate-lyase activity. Unfortunately, none of the primers used to detect the presence of genes encoding polysaccharide-degrading enzymes were successfully amplified.

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Molecular characterization of three Hsp90 from Pieris and expression patterns in response to cold and thermal stress in summer and winter diapause of Pieris melete

Zou

et al. 2017

Insect Science

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Heat shock proteins (Hsps) have been linked to stresses and winter diapause in insects, but whether they are components of summer diapause is still unknown. In this study, complementary DNAs of Hsp90 from Pieris melete, Pieris rapae and Pieris canidia named PmHsp90, PrHsp90 and PcHsp90, respectively, were cloned and sequenced. The deduced amino acid sequence consisted of 718 amino acid residues with a putative molecular mass of 82.6, 82.6 and 82.7 kDa, respectively. The amino acid sequences contained all of the five conserved signature motifs in the Hsp90 family and a bHLH protein folding activity region. The differential expression pattern of PmHsp90 in response to summer diapause and winter diapause, which are related to heat/cold stress, was investigated. Cold stress induced Hsp90 up-regulation in summer and winter diapause pupae, but not in non-diapause individuals. Heat shock up-regulated PmHsp90 gradually with an increase in temperature in summer diapause, and PmHsp90 was rapidly up-regulated in winter diapause. After 30 min heat shock at 39°C, substantial up-regulation of PmHsp90 transcript levels were observed both in summer and winter diapause. However, in non-diapause a relatively stable expression was found under different durations of 39°C heat shock. Compared to the optimal treatment of 18°C for diapause development, a high temperature acclimation of 31°C induced PmHsp90 up-regulation in summer diapause, whereas a low temperature acclimation of 4°C induced up-regulation in winter diapause. The current results indicate that Hsp90 may play an important role in response to heat/cold stress both in summer and winter diapause.

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Universal primers for amplifying the complete coding sequence of cytoplasmic heat shock protein 90 (HSP90) in Lepidoptera

Cited by 4 publications

References 29 publications

More than a colour change: insect melanism, disease resistance and fecundity

More than a colour change: insect melanism, disease resistance and fecundity

First report of seaweed-associated yeast from Indonesia: Species composition and screening of their polysaccharides-degrading enzymes

Molecular characterization of three Hsp90 from Pieris and expression patterns in response to cold and thermal stress in summer and winter diapause of Pieris melete

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