Abbreviations: ACC, acetyl-CoA carboxylase; AMP, adenosine monophosphate; AMPK, adenosine monophosphate activated protein kinase; CCCP, carbonyl cyanide m-chlorophenyl hydrazine; CISD1, CDGSH iron sulfur domain 1; DRP1, dynamin-related protein 1; GFP, green fluorescence protein; MFF, mitochondrial fission factor; MFN-1/2, mitofusin-1/2; mtFIS1 101-152 , mitochondrial targeting sequence of FIS1 (amino acids 101-152); NDP52, nuclear dot protein 52; OPA1, dynamin-like 120 kDa protein; OPTN, optineurin; OXPHOS, oxidative phosphorylation; PINK1, PTEN-induced kinase 1; SQSTM1/p62, sequestosome-1; TBK1, TANK-binding kinase 1; Ub, ubiquitin; UBA UBQLN1 , his-halo-ubiquilin1 UBA domain tetramer; ULK1, unc-51 like autophagy activating kinase 1. AbstractMitophagy is a key process regulating mitochondrial quality control. Several mechanisms have been proposed to regulate mitophagy, but these have mostly been studied using stably expressed non-native proteins in immortalized cell lines. In skeletal muscle, mitophagy and its molecular mechanisms require more thorough investigation. To measure mitophagy directly, we generated a stable skeletal muscle C2C12 cell line, expressing a mitophagy reporter construct (mCherry-green fluorescence protein-mtFIS1 101-152 ). Here, we report that both carbonyl cyanide m-chlorophenyl hydrazone (CCCP) treatment and adenosine monophosphate activated protein kinase (AMPK) activation by 991 promote mitochondrial fission via phosphorylation of MFF and induce mitophagy by ~20%. Upon CCCP treatment, but not 991, ubiquitin phosphorylation, a read-out of PTEN-induced kinase 1 (PINK1) activity, and Parkin E3 ligase activity toward CDGSH iron sulfur domain 1 (CISD1) were increased. Although the PINK1-Parkin signaling pathway is active in response to CCCP treatment, we observed no change in markers of mitochondrial protein content. Interestingly, our data shows that TANK-binding kinase 1 (TBK1) phosphorylation is increased after both CCCP and 991 treatments, suggesting TBK1 activation to be independent of both PINK1 and Parkin. Finally, we confirmed in non-muscle cell lines that TBK1 phosphorylation occurs in the absence of PINK1 and is regulated by AMPKdependent signaling. Thus, AMPK activation promotes mitophagy by enhancing mitochondrial fission (via MFF phosphorylation) and autophagosomal engulfment (via TBK1 activation) in a PINK1-Parkin independent manner.
Currently, the global agricultural system is focused on a limited number of crop species, thereby presenting a threat to food security and supply, especially with predicted global climate change conditions. The importance of ‘underutilized’ crop species in meeting the world’s demand for food has been duly recognized by research communities, governments and policy makers worldwide. The development of underutilized crops, with their vast genetic resources and beneficial traits, may be a useful step towards solving food security challenges by offering a multifaceted agricultural system that includes additional important food resources. Bambara groundnut is among the beneficial underutilized crop species that may have a positive impact on global food security through organized and well-coordinated multidimensional breeding programs. The excessive degrees of allelic difference in Bambara groundnut germplasm could be exploited in breeding activities to develop new varieties. It is important to match recognized breeding objectives with documented diversity in order to significantly improve breeding. This review assesses the genetic diversity of Bambara groundnut, as well as important factors involved in realizing and harnessing the potential of this crop.
Knowledge of agro-morphological genetic variation and cropping conditions on vegetative and yield-related traits plays a significant role in varietal improvement and production of eggplant (Solanum melongena L.). Following this premise, the current study was conducted to critically asses the genetic variation of 29 eggplant accessions by using agro-morphological characterization evaluated under two cropping conditions, namely, glasshouse and open field. The experiments were laid out in randomized complete block design (RCBD) with three replications. Data on vegetative and yield characteristics were collected and subjected to analysis of variance (ANOVA) using SAS 9.4, while variance components were estimated manually. The results obtained from the analysis of variance indicated a highly significant difference (p ≤ 0.01) for all characteristics studied in both cropping conditions. The evaluated accessions were grouped into six major clusters based on agro-morphological traits using Unweighted Pair Group Method with Arithmetic mean (UPGMA) dendrogram. Hence, crosses between group I with VI or V could be used to attain higher heterosis and vigor among the accessions. Also, this evaluation could be used as a selection criterion for important yield agronomic traits in eggplant. The methodology and the approaches used may provide a model for the enhancement of other vegetable crop diversity towards adaptability to the cropping condition decision. This result displayed importance for preserving eggplant germplasm for future varietal development and revealed that open field cropping condition is more suitable under Malaysia’s agroecology.
Bacterial leaf blight caused by Xanthomonas oryzae pv oryzae (Xoo) and blast caused by Magnaporthe oryzae are major diseases responsible for significant yield loss in rice production across all rice growing regions. Host plant resistance has been advocated as a sustainable means of guarding against the diseases. This experiment was conducted with the aim to introgress multiple resistance genes against bacterial leaf blight and blast diseases through marker-assisted backcross breeding. Two dominant (Xa4 and Xa21) and two recessive (xa5 and xa13) Xoo resistance genes were introgressed into a high yielding Malaysian rice variety Putra-1 with genetic background of three blast resistance (Piz, Pi2 and Pi9) genes. Eight polymorphic tightly linked functional and SSR markers were used for foreground selection of target genes. Seventy nine polymorphic SSR markers were used in background selection. The plants were challenged at initial stage of breeding and challenged again at BC2F2 with the most virulent Malaysian pathotypes of Xoo (P7.7) and Magnaporthe oryzae (P7.2) to test their resistance. Results obtained from foreground marker segregation analysis at BC1F1 and BC2F1 showed that the marker polymorphism both fitted into the Mendel’s single gene segregation ratio of 1:1 for both Xoo and blast resistance. At BC2F2, results indicated that foreground marker polymorphism fitted into the expected Mendelian ratio of 1:2:1 for blast resistance only. Marker-assisted background selection revealed high percentage of recurrent parent genome recovery (95.9%). It was concluded that the inheritance of blast resistance in the introgressed lines was mainly due to single gene action while the inheritance of Xoo resistance was substantially due to single nuclear gene action. The incorporation of four bacterial leaf blight and three blast resistance genes (Xa4 + xa5 + xa13 + Xa21; Pi9 + Pi2 + Piz) in the newly developed lines would provide for broad spectrum and durable resistance against the two major diseases studied.
Tocotrienol-rich fraction (TRF) is an extract of palm oil that consists of 25% α-tocopherol and 75% tocotrienols. TRF was shown to possess antioxidant, anti-inflammatory, anticancer, neuroprotective and cholesterol-lowering activities. Glutamate is the major mediator of excitatory signals in the mammalian central nervous system. Extreme amounts of glutamate in the extracellular spaces can lead to numerous neurodegenerative diseases. Hence, the efficacy of oil palm TRF and α-tocopherol in protecting astrocytes against glutamate-induced cell death was studied. Specifically, the effectiveness of pre-or post-treatment of TRF and αtocopherol upon glutamate excitotoxicity was determined by evaluating cell viability and morphology of astrocytes. Cell viability was measured using MTT assay while cell morphology was monitored under fluorescent microscope using the acridine orange/propidium iodide (AO/PI) assay. Exposure to 230 mM glutamate significantly reduced cell viability to 50% in both the pre-and post-treatment studies; however, pre-and post-treatment with TRF and α-tocopherol attenuated the cytotoxic effect of glutamate. Compared to glutamate-injured astrocytes, pre-treatment with 100, 200 and 300 ng/ml TRF significantly improved cell viability following glutamate injury to 86.6%, 86.7% and 93.9%, respectively (p < 0.05). On the contrary, high concentrations of α-tocopherol promote cell death. This study shows that TRF not only provide a better protection against glutamate toxicity (pre-treatment), but was also able to reverse the lipid peroxidation resulting from glutamate insults (post-treatment). The present results demonstrate that TRF, but not αtocopherol, protected the astrocytes against glutamate-induced cell death, indicating its neuro-protective potential.
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