The Landscape of Autophagy-Related (ATG) Genes and Functional Characterization of TaVAMP727 to Autophagy in Wheat

Autophagy is an indispensable biological process and plays crucial roles in plant growth and plant responses to both biotic and abiotic stresses. This study systematically identified autophagy-related proteins (ATGs) in wheat and its diploid and tetraploid progenitors and investigated their genomic organization, structure characteristics, expression patterns, genetic variation, and regulation network. We identified a total of 77, 51, 29, and 30 ATGs in wheat, wild emmer, T. urartu and A. tauschii, respectively, and grouped them into 19 subfamilies. We found that these autophagy-related genes (ATGs) suffered various degrees of selection during the wheat’s domestication and breeding processes. The genetic variations in the promoter region of Ta2A_ATG8a were associated with differences in seed size, which might be artificially selected for during the domestication process of tetraploid wheat. Overexpression of TaVAMP727 improved the cold, drought, and salt stresses resistance of the transgenic Arabidopsis and wheat. It also promoted wheat heading by regulating the expression of most ATGs. Our findings demonstrate how ATGs regulate wheat plant development and improve abiotic stress resistance. The results presented here provide the basis for wheat breeding programs for selecting varieties of higher yield which are capable of growing in colder, drier, and saltier areas.

Genome-Wide Characterization of Sedum plumbizincicola HMA Gene Family Provides Functional Implications in Cadmium Response

Heavy-metal ATPase (HMA), an ancient family of transition metal pumps, plays important roles in the transmembrane transport of transition metals such as Cu, Zn, Cd, and Co. Although characterization of HMAs has been conducted in several plants, scarcely knowledge was revealed in Sedum plumbizincicola, a type of cadmium (Cd) hyperaccumulator found in Zhejiang, China. In this study, we first carried out research on genome-wide analysis of the HMA gene family in S. plumbizincicola and finally identified 8 SpHMA genes and divided them into two subfamilies according to sequence alignment and phylogenetic analysis. In addition, a structural analysis showed that SpHMAs were relatively conserved during evolution. All of the SpHMAs contained the HMA domain and the highly conserved motifs, such as DKTGT, GDGxNDxP, PxxK S/TGE, HP, and CPx/SPC. A promoter analysis showed that the majority of the SpHMA genes had cis-acting elements related to the abiotic stress response. The expression profiles showed that most SpHMAs exhibited tissue expression specificity and their expression can be regulated by different heavy metal stress. The members of Zn/Co/Cd/Pb subgroup (SpHMA1-3) were verified to be upregulated in various tissues when exposed to CdCl2. Here we also found that the expression of SpHMA7, which belonged to the Cu/Ag subgroup, had an upregulated trend in Cd stress. Overexpression of SpHMA7 in transgenic yeast indicated an improved sensitivity to Cd. These results provide insights into the evolutionary processes and potential functions of the HMA gene family in S. plumbizincicola, laying a theoretical basis for further studies on figuring out their roles in regulating plant responses to biotic/abiotic stresses.

Prospects for Knockout of MYB60, a Transcriptional Repressor of Anthocyanin Biosynthesis, in Brassicaceae Plants by Genome Editing

The Brassicaceae plant family contains many economically important crops such as Brassica napus L., Brassica rapa L., Brassica oleracea L., Brassica juncea L., Eruca sativa Mill., Camelina sativa L. and Raphanus sativus L. Insufficient data on the genetic regulation of agronomic traits in these species complicates the editing of their genomes. In recent years, the attention of the academic community has been drawn to anthocyanin hyperaccumulation. This trait is not only beneficial for human health, but can also increase plant resistance to stress. MYB transcription factors are the main regulators of flavonoid biosynthesis in plants. Some of them are well studied in Arabidopsis thaliana. The AtMYB60 gene is a transcriptional repressor of anthocyanin biosynthesis, and it also negatively impacts plant responses to drought stress. Myb60 is one of the least studied transcription factors with similar functions in Brassicaceae. There is a high degree of homology between predicted MYB60 genes of A. thaliana and related plant species. However, functions of these homologous genes have never been studied. Gene knockout by CRISPR/Cas technology remains the easiest way to perform genome editing in order to discover the role of individual plant genes. Disruption of genes acting as negative regulators of anthocyanin biosynthesis could result in color staining of plant tissues and an increase in stress tolerance. In the present study, we investigated the AtMYB60 gene and its homologs in Brassicaceae plants and suggested universal gRNAs to knockout these genes. Keywords: CRISPR, Brassicaceae, MYB60, knockout, anthocyanin

Analysis of the transcriptome of the needles and bark of Pinus radiata induced by bark stripping and methyl jasmonate

Background Plants are attacked by diverse insect and mammalian herbivores and respond with different physical and chemical defences. Transcriptional changes underlie these phenotypic changes. Simulated herbivory has been used to study the transcriptional and other early regulation events of these plant responses. In this study, constitutive and induced transcriptional responses to artificial bark stripping are compared in the needles and the bark of Pinus radiata to the responses from application of the plant stressor, methyl jasmonate. The time progression of the responses was assessed over a 4-week period. Results Of the 6312 unique transcripts studied, 86.6% were differentially expressed between the needles and the bark prior to treatment. The most abundant constitutive transcripts were related to defence and photosynthesis and their expression did not differ between the needles and the bark. While no differential expression of transcripts were detected in the needles following bark stripping, in the bark this treatment caused an up-regulation and down-regulation of genes associated with primary and secondary metabolism. Methyl jasmonate treatment caused differential expression of transcripts in both the bark and the needles, with individual genes related to primary metabolism more responsive than those associated with secondary metabolism. The up-regulation of genes related to sugar break-down and the repression of genes related with photosynthesis, following both treatments was consistent with the strong down-regulation of sugars that has been observed in the same population. Relative to the control, the treatments caused a differential expression of genes involved in signalling, photosynthesis, carbohydrate and lipid metabolism as well as defence and water stress. However, non-overlapping transcripts were detected between the needles and the bark, between treatments and at different times of assessment. Methyl jasmonate induced more transcriptional responses in the bark than bark stripping, although the peak of expression following both treatments was detected 7 days post treatment application. The effects of bark stripping were localised, and no systemic changes were detected in the needles. Conclusion There are constitutive and induced differences in the needle and bark transcriptome of Pinus radiata. Some expression responses to bark stripping may differ from other biotic and abiotic stresses, which contributes to the understanding of plant molecular responses to diverse stresses. Whether the gene expression changes are heritable and how they differ between resistant and susceptible families identified in earlier studies needs further investigation.

Find New RoadsTM? A Systematic Review on the Impacts of Off-Road Vehicle Activity on Soil, Vegetation, and Wildlife

The global use of off-road vehicles (ORVs) in natural environments has accelerated rapidly over the last few decades, resulting in significant social and environmental consequences. As the demand, use, and promotion of light-duty ORVs like all-terrain vehicles (ATVs), motorcycles, four-wheel drive trucks and sport utility vehicles (SUVs) increases in remote wilderness, the landscape is becoming fragmented into disorganized and destructive networks of trails and roads. Substantial ecological impacts to a wide range of ecosystem structures and functions will likely result from ORV activity. Applying a global systematic review, we examine 105 publications about plant, soil, and wildlife responses to ORV traffic in different habitats to help guide the direction of future research, monitoring programs, and mitigation efforts. Most studies investigated impacts to animals, followed by soils, then vegetative responses. Soil studies primarily focused on physical impacts to the soil (i.e., compaction, erosion, rut depth), but some studies suggest that soil chemical and biological properties may also be impacted by ORV traffic. The literature on plant responses to ORV activities primarily explored vegetation loss, although impacts on the plant community were also investigated. Animal studies investigated impacts of ORV use on invertebrates, mammals, birds, and to a lesser extent reptiles/amphibians, including population-level, community-level, and behavioral responses. Overall, research on environmental impacts of ORV traffic is biased to coastal and desert ecosystems in the northern hemisphere (primarily in the US), often does not address mechanisms that may produce ecological impacts (e.g., intensity of vehicular disturbance and ecosystem- or species-specific sensitivity to ORV activities), and frequently focused on short-term responses. More research is needed to understand the mechanisms that cause the different responses of soil, plant, and animals to ORVs over the long-term in a broad range of ecosystems to support real-time management and conservation efforts.

Down-Regulation of Cytokinin Receptor Gene SlHK2 Improves Plant Tolerance to Drought, Heat, and Combined Stresses in Tomato

Environmental stresses negatively affect the growth and development of plants. Several previous studies have elucidated the response mechanisms of plants to drought and heat applied separately; however, these two abiotic stresses often coincide in environmental conditions. The global climate change pattern has projected that combined drought and heat stresses will tend to increase in the near future. In this study, we down-regulated the expression of a cytokinin receptor gene SlHK2 using RNAi and investigated the role of this gene in regulating plant responses to individual drought, heat, and combined stresses (drought + heat) in tomato. Compared to the wild-type (WT), SlHK2 RNAi plants exhibited fewer stress symptoms in response to individual and combined stress treatments. The enhanced abiotic stress tolerance of SlHK2 RNAi plants can be associated with increased membrane stability, osmoprotectant accumulation, and antioxidant enzyme activities. Furthermore, photosynthesis machinery was also protected in SlHK2 RNAi plants. Collectively, our results show that down-regulation of the cytokinin receptor gene SlHK2, and consequently cytokinin signaling, can improve plant tolerance to drought, heat, and combined stress.

Soil and Plant Responses to Phosphorus Inputs from Different Phytase-Associated Animal Diets

The over-supplementation of animal feeds with phosphorus (P) within livestock-production systems leads to high rates of P excretion and thus to high P loads and losses, which negatively impact the natural environment. The addition of phytase to pig and poultry diets can contribute to reducing P excretion; however, cascading effects of phytase on plant–soil systems remain poorly understood. Here, we addressed how three different diets containing various levels of exogenous phytase, i.e., (1) no-phytase, (2) phytase (250 FTU), and (3) superdose phytase (500 FTU) for pigs (Sus scrofa domesticus) and broilers (Gallus gallus domesticus) might affect P dynamics in two different plant–soil systems including comfrey (Symphytum ×uplandicum) and ryegrass (Lolium perenne). We found that differences in phytase supplementation significantly influenced total P content (%) of broiler litter and also pig slurry (although not significantly) as a result of dietary P content. P Use Efficiency (PUE) of comfrey and ryegrass plants was significantly higher under the intermediate ‘phytase’ dose (i.e., commercial dose of 250 FTU) when compared to ‘no-phytase’ and ‘superdose phytase’ associated with pig slurry additions. Soil P availability (i.e., water soluble P, WSP) in both comfrey and ryegrass mesocosms significantly decreased under the intermediate ‘phytase’ treatment following pig slurry additions. Dietary P content effects on P losses from soils (i.e., P leaching) were variable and driven by the type of organic amendment. Our study shows how commercial phytase levels together with higher dietary P contents in pig diets contributed to increase PUE and decrease WSP thus making the plant–soil system more P conservative (i.e., lower risks of P losses). Our evidence is that dietary effects on plant–soil P dynamics are driven by the availability of P forms (for plant uptake) in animal excretes and the type of organic amendment (pig vs. broiler) rather than plant species identity (comfrey vs. ryegrass).

The Evolution and Functional Roles of miR408 and Its Targets in Plants

MicroRNA408 (miR408) is an ancient and highly conserved miRNA, which is involved in the regulation of plant growth, development and stress response. However, previous research results on the evolution and functional roles of miR408 and its targets are relatively scattered, and there is a lack of a systematic comparison and comprehensive summary of the detailed evolutionary pathways and regulatory mechanisms of miR408 and its targets in plants. Here, we analyzed the evolutionary pathway of miR408 in plants, and summarized the functions of miR408 and its targets in regulating plant growth and development and plant responses to various abiotic and biotic stresses. The evolutionary analysis shows that miR408 is an ancient and highly conserved microRNA, which is widely distributed in different plants. miR408 regulates the growth and development of different plants by down-regulating its targets, encoding blue copper (Cu) proteins, and by transporting Cu to plastocyanin (PC), which affects photosynthesis and ultimately promotes grain yield. In addition, miR408 improves tolerance to stress by down-regulating target genes and enhancing cellular antioxidants, thereby increasing the antioxidant capacity of plants. This review expands and promotes an in-depth understanding of the evolutionary and regulatory roles of miR408 and its targets in plants.