A load spectra design method for multi-stress accelerated testing

Accelerated testing (AT) is an effective testing method for evaluating the reliability of highly reliable products. In many applications, such products are often subject to multiple stresses in actual working environments. Then, how to design the load spectra for multi-stress AT becomes a critical issue. In this paper, a new load spectra design method is proposed to overcome this challenge. Considering the properties of multiple stresses, a novel multi-stress loading matrix (MSLM) is modified to fully describe the load spectra in actual working environments and to generate the corresponding load spectra for AT. For illustration purposes, an inverse Gaussian (IG) process incorporating life-stress relationships is adopted to model product degradation under multiple stresses. A case study on a type of hydraulic piston pump is provided to demonstrate the use of the proposed method in guiding the load spectra design for multi-stress AT.

Histone Acetyltransferases and Deacetylases Are Required for Virulence, Conidiation, DNA Damage Repair, and Multiple Stresses Resistance of Alternaria alternata

Histone acetylation, which is critical for transcriptional regulation and various biological processes in eukaryotes, is a reversible dynamic process regulated by HATs and HDACs. This study determined the function of 6 histone acetyltransferases (HATs) (Gcn5, RTT109, Elp3, Sas3, Sas2, Nat3) and 6 histone deacetylases (HDACs) (Hos2, Rpd3, Hda1, Hos3, Hst2, Sir2) in the phytopathogenic fungus Alternaria alternata by analyzing targeted gene deletion mutants. Our data provide evidence that HATs and HDACs are both required for mycelium growth, cell development and pathogenicity as many gene deletion mutants (ΔGcn5, ΔRTT109, ΔElp3, ΔSas3, ΔNat3, ΔHos2, and ΔRpd3) displayed reduced growth, conidiation or virulence at varying degrees. In addition, HATs and HDACs are involved in the resistance to multiple stresses such as oxidative stress (Sas3, Gcn5, Elp3, RTT109, Hos2), osmotic stress (Sas3, Gcn5, RTT109, Hos2), cell wall-targeting agents (Sas3, Gcn5, Hos2), and fungicide (Gcn5, Hos2). ΔGcn5, ΔSas3, and ΔHos2 displayed severe growth defects on sole carbon source medium suggesting a vital role of HATs and HDACs in carbon source utilization. More SNPs were generated in ΔGcn5 in comparison to wild-type when they were exposed to ultraviolet ray. Moreover, ΔRTT109, ΔGcn5, and ΔHos2 showed severe defects in resistance to DNA-damaging agents, indicating the critical role of HATs and HDACs in DNA damage repair. These phenotypes correlated well with the differentially expressed genes in ΔGcn5 and ΔHos2 that are essential for carbon sources metabolism, DNA damage repair, ROS detoxification, and asexual development. Furthermore, Gcn5 is required for the acetylation of H3K4. Overall, our study provides genetic evidence to define the central role of HATs and HDACs in the pathological and biological functions of A. alternata.

Physiological and molecular mechanism of tolerance of two maize genotypes under multiple abiotic stresses.

Abiotic stresses are the major threat to crops regardless of their nature, duration, and frequency, their occurrence either singly, and or combination is deleterious for the plant growth and development. Maize is the most important crop largely grown in the tropical regions in the summer rainy season, often facing a stress combination of drought and waterlogging. We previously showed under multiple stresses up-regulated leaf proteins of maize plants were involved to enhance the tolerance mechanism of tolerant genotype. Whereas, in susceptible genotypes up-regulated proteins ameliorate to survive the stressful condition. Further to understand the response of roots proteome under multiple stresses was determined using the 2DE technique. The results of the root proteome show the up-regulated proteins of CML49 genotype (tolerant) are involved in enhancing the N content, cell wall remodeling, and acclimatization during the stresses. Up-regulated proteins of CML100 genotype (sensitive) are stressed markers of roots' primary and secondary metabolism. However, the root proteome of both genotypes correlates with the leaf proteome (previous). Therefore, the present study and our previous results provide comprehensive insight into the molecular mechanisms of tolerance in multiple abiotic stresses of maize plants.

Phenotypic and transcriptomic responses of cultivated sunflower seedlings (Helianthus annuus L.) to four abiotic stresses

Plants encounter and respond to numerous abiotic stresses during their lifetimes. These stresses are often related and could therefore elicit related responses. There are, however, relatively few detailed comparisons between multiple different stresses at the molecular level. Here, we investigated the phenotypic and transcriptomic response of cultivated sunflower (Helianthus annuus L.) seedlings to three water-related stresses (i.e., dry-down, an osmotic challenge with polyethylene glycol 6000 [PEG], and salt stress), as well as a generalized low-nutrient stress. Our goal was to identify commonalities in the response to the three water-related stresses and compare them to a distinct low-nutrient stress. All four stresses negatively impacted seedling growth, with the low-nutrient stress having a more divergent response from control as compared to the water-related stresses. Observed phenotypic responses were consistent with expectation for growth in low-resource environments, including increased (i.e., less negative) carbon fractionation values and leaf C:N ratios, as well as increased belowground biomass allocation. Analysis of the leaf and root transcriptome under each stress scenario revealed that most genes were differentially expressed in response to multiple stresses. The number of differentially expressed genes (DEGs) under stress was greater in leaf tissue, but roots exhibited a higher proportion of DEGs unique to individual stresses. Overall, the three water-related stresses had a more similar transcriptomic response to each other vs. low-nutrient stress, though this pattern was more pronounced in root tissue than in leaf tissue. In contrast with the results of our differential expression analysis, co-expression network analysis revealed that the response to each of the four stresses in our study were generally non-overlapping and there was little indication of a shared co-expression response despite the majority of DEGs being shared between multiple stresses. Importantly, PEG stress, which is often used to simulate drought stress in experimental settings, had little transcriptomic resemblance to true water limitation (i.e., dry-down) in our study calling into question its utility as a means for simulating drought.

Selection and Optimization of Reference Genes for MicroRNA Expression Normalization by qRT-PCR in Chinese Cedar (Cryptomeria fortunei) under Multiple Stresses

MicroRNA (miRNA) expression analysis is very important for investigating its functions. To date, no research on reference genes (RGs) for miRNAs in gymnosperms, including Cryptomeria fortunei, has been reported. Here, ten miRNAs (i.e., pab-miR159a, cln-miR162, cas-miR166d, pab-miR395b, ppt-miR894, cln-miR6725, novel1, novel6, novel14 and novel16) and three common RGs (U6, 5S and 18S) were selected as candidate RGs. qRT-PCR was used to analyse their expressions in C. fortunei under various experimental conditions, including multiple stresses (cold, heat, drought, salt, abscisic acid and gibberellin) and in various tissues (roots, stems, tender needles, cones and seeds). Four algorithms (delta Ct, geNorm, NormFinder and BestKeeper) were employed to assess the stability of candidate RG expression; the geometric mean and RefFinder program were used to comprehensively evaluate RG stability. According to the results, novel16, cln-miR6725, novel1 and U6 were the most stable RGs for studying C. fortunei miRNA expression. In addition, the expression of three target miRNAs (aly-miR164c-5p, aly-miR168a-5p and smo-miR396) was examined to verify that the selected RGs are suitable for miRNA expression normalisation. This study may aid further investigations of miRNA expression/function in the response of C. fortunei to abiotic stress and provides an important basis for the standardisation of miRNA expression in other gymnosperm species.