Identifying the functional groups and the influence of synthetic chelators on Cd availability and microbial biomass carbon in Cd-contaminated soil

Synthetic chelators play an important role in boosting the microbial biomass carbon (MBC), dissolved organic carbon (DOC), and heavy metal solubility in a contaminated soil toward a sustainability of environment for agricultural crops. Castor plant was grown under different levels of Cd contaminated soil (?Cd and +Cd) following adding three chelating agents, ethylenediaminetetraacetic acid (H₄EDTA), nitriloacetic acid (H₃ NTA), and NH₄ citrate (ammonium citrate) to the soil at rates of 10, 15, and 25 mmol in 5 kg of soil per pot. The highest bioavailable Cd concentrations in soil and castor plant were obtained from NH₄ citrate and H₄EDTA treatments in the contaminated soil. Fourier transform infrared (FTIR) analysis showed that NH₄ citrate was the most effective chelator in Cd-contaminated soil. MBC and DOC contents were significantly increased and reached at 81.98–80.37 and 1.96–1.90 mg kg^?1 respectively, in the (H₃ NTA) and NH₄ citrate treatments in Cd-contaminated soil. Further research is needed to investigate the use of chelators in the phytoextraction of Cd-contaminated soils under field conditions and whether it may be beneficial in accelerating the phytoextraction of Cd through hyperaccumulating plants.     

Identification of small molecule inhibitors of ALK2: a virtual screening,density functional theory,and molecular dynamics simulations study

Bone morphogenetic proteins (BMPs) are a family of more than 30 ligands and several receptors, such as activin like kinases (ALKs) and bone morphogenetic protein receptor (BMPR). Physiological significance of these proteins lies in their prominent role during homeostasis, apoptosis, tissue remodeling, embryonic patterning, and normal development. Fibrodysplasia ossificans progressive (FOP) is one among several other diseases caused by impaired BMP signaling. FOP is caused by the pathogenicity of activating mutation of ALK2. In order to treat FOP, a search for good inhibitors of ALK2 based on dorsomorphin and LDN substitution, which in essence is a ligand based search of inhibitors, is in progress. Contributing to this area of research we identified several lead molecules based on protein structure using virtual screening. After virtual screening of a huge library of small molecules and ab initio calculation of selected molecules for drug efficacy, we did molecular dynamic simulation of lead molecules and protein complexes. We identified five potential drug molecules that show very stable binding on the same binding site as LDN-213844. We also ranked these lead molecules based on MM/PBSA binding energy. This study provides a basis to think beyond the pyrimidine nucleus of dorsomorphin/LDN and design new chemical derivatives for effective treatment of FOP.

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Graphical abstract Small molecule inhibitors of ALK2

     

Precise editing of CLAVATA genes in Brassica napus L. regulates multilocular silique development

Multilocular silique is a desirable agricultural trait with great potential for the development of high‐yield varieties of Brassica. To date, no spontaneous or induced multilocular mutants have been reported in Brassica napus, which likely reflects its allotetraploid nature and the extremely low probability of the simultaneous random mutagenesis of multiple gene copies with functional redundancy. Here, we present evidence for the efficient knockout of rapeseed homologues of CLAVATA3 (CLV3) for a secreted peptide and its related receptors CLV1 and CLV2 in the CLV signalling pathway using the CRISPR/Cas9 system and achieved stable transmission of the mutations across three generations. Each BnCLV gene has two copies located in two subgenomes. The multilocular phenotype can be recovered only in knockout mutations of both copies of each BnCLV gene, illustrating that the simultaneous alteration of multiple gene copies by CRISPR/Cas9 mutagenesis has great potential in generating agronomically important mutations in rapeseed. The mutagenesis efficiency varied widely from 0% to 48.65% in T₀ with different single‐guide RNAs (sgRNAs), indicating that the appropriate selection of the sgRNA is important for effectively generating indels in rapeseed. The double mutation of BnCLV3 produced more leaves and multilocular siliques with a significantly higher number of seeds per silique and a higher seed weight than the wild‐type and single mutant plants, potentially contributing to increased seed production. We also assessed the efficiency of the horizontal transfer of Cas9/gRNA cassettes by pollination. Our findings reveal the potential for plant breeding strategies to improve yield traits in currently cultivated rapeseed varieties.     

Serum organochlorine pesticides residues and risk of cancer: A case-control study

Organochlorine pesticides (OCPs) are frequently used worldwide as insecticides, fungicides, herbicides and termiticides and have been associated with a variety of cancers in animal and human studies. In the present study, we examined residues of fourteen OCPs in the serum samples of diagnosed cancer patients and healthy residents of Karachi, Pakistan. A random collection of fasting blood samples was carried out from the donors with informed consent. Serum was separated within 2?h of blood collection and was then subjected to extraction with organic solvents followed by purification with florisil column. The final organic extract of each serum sample was processed with Gas Chromatograph coupled with Electron Capture Detector (GC-ECD). OCPs were detected in 97.59% of the cancer cases and 93.75% of the healthy subjects. Mean concentrations of total OCPs (ΣOCPs) was found elevated in the cancer group (0.606?mg/kg) compared with the control group (0.322?mg/kg). Endosulfan was the highest prevalent OCP with a mean concentration of 0.214?mg/kg in the cancer group and 0.166?mg/kg in the control group. The second most prevalent OCP was 4,4-DDE with a mean concentration of 0.131?mg/kg in the cancer group and 0.019?mg/kg in the control group. Highest level of ΣOCPs was detected in the breast cancer cases (20.411?mg/kg) with a mean level of (2.041?mg/kg). In light of the obtained results and available literature on the subject, it has been concluded that OCPs are positively associated with the risk of various cancers in humans.     

Arabidopsis HIPP27 is a host susceptibility gene for the beet cyst nematode Heterodera schachtii

Sedentary plant‐parasitic cyst nematodes are obligate biotrophs that infect the roots of their host plant. Their parasitism is based on the modification of root cells to form a hypermetabolic syncytium from which the nematodes draw their nutrients. The aim of this study was to identify nematode susceptibility genes in Arabidopsis thaliana and to characterize their roles in supporting the parasitism of Heterodera schachtii. By selecting genes that were most strongly upregulated in response to cyst nematode infection, we identified HIPP27 (HEAVY METAL‐ASSOCIATED ISOPRENYLATED PLANT PROTEIN 27) as a host susceptibility factor required for beet cyst nematode infection and development. Detailed expression analysis revealed that HIPP27 is a cytoplasmic protein and that HIPP27 is strongly expressed in leaves, young roots and nematode‐induced syncytia. Loss‐of‐function Arabidopsis hipp27 mutants exhibited severely reduced susceptibility to H. schachtii and abnormal starch accumulation in syncytial and peridermal plastids. Our results suggest that HIPP27 is a susceptibility gene in Arabidopsis whose loss of function reduces plant susceptibility to cyst nematode infection without increasing the susceptibility to other pathogens or negatively affecting the plant phenotype.     

RNAi-mediated silencing of endogenous <Emphasis Type="Italic">Vlnv</Emphasis> gene confers stable reduction of cold-induced sweetening in potato (<Emphasis Type="Italic">Solanum tuberosum</Emphasis> L. cv. Désirée)

Potato tubers must be cold-stored to extend their shelf life and maintain an uninterrupted supply chain for food processors. However, a side-effect of low-temperature storage is manifested in terms of cold-induced sweetening (CIS) of potato tubers, which reduces the processing quality and the commercial value of the end-products. RNA interference (RNAi) technology, whereby transgene-derived small interfering RNAs can trigger the homology-based knockdown of cognate host genes and can initiate gene silencing, has been successfully applied in crop improvement through targeted gene knockout in host plants. In the current study, transgenic potato plants (Solanum tuberosum cv. Désirée) were generated, expressing a 300 bp hairpin loop nucleotide sequence targeting the potato vacuolar invertase gene (VInv), under the constitutive Cauliflower mosaic virus 35S promoter. Tubers collected from transgenic lines showed a significant reduction in reducing sugar content after 180 days of cold storage, without showing any measurable off-target effects on plant morphology and tuberization compared to non-transformed control plants. The cold-stored tubers were further assayed for chip color, which showed a fairly light colored quality in the samples originating from RNAi lines. Together with similar effects seen in previously published experiments involving other potato varieties, the Désirée results described here establish the efficacy of using RNAi for the successful reduction of CIS in potato tubers.     

Earliness index deters <Emphasis Type="Italic">Pectinophora gossypiella</Emphasis> incidence on advanced cultivars of <Emphasis Type="Italic">Bt</Emphasis> cotton

Pink bollworm (Pectinophora gossypiella) is recognized as an important pest of cotton and can damage flowers and bolls of both Bt and non-Bt cultivars. Cry-1Ac in Bt cultivars is considered very effective in controlling lepidopterous larvae; therefore, the present study was carried out to investigate the impact of Cry1-Ac and the earliness index on the natural incidence of P. gossypiella at the Cotton Research Institute, Faisalabad. During 2015–2016, ten cultivars were used to determine the incidence of pink bollworm infestation. The experiment was repeated for 2 years. During the next year, Cry1-Ac and earliness traits of selected cultivars were also observed to determine their impact on pink bollworm. Correlation coefficient results regarding days to first flower (r value = 0.66) as well as the earliness index (r value = ? 0.62) exhibited a strong association with pink bollworm, but Cry1-Ac had a weak association (r value = ? 0.058) with pink bollworm. The coefficient of determination (R ²) explained that variability of pink bollworm due to Cry1-Ac, the earliness index, and days to first flower was 18.0, 38.5, and 43.5%, respectively. Principal component analysis results showed that the first two PCs expressed 87% of the total variability. Clusters made on the basis of the studied parameters revealed that clusters 2 and 3 comprised the cotton cultivars possessing earliness traits compared with cluster 1. Therefore, it can be concluded that the earliness index in cotton is an important component for the sustainable management of pink bollworm infestation, the need for which is endless to evade the pink bollworm problem in the era of climate change.     

Secondary metabolism of pharmaceuticals in the plant in vitro cultures: strategies,approaches, and limitations to achieving higher yield

Biotechnology is playing a vital alternative role in the production of pharmaceutical plant secondary metabolites to support industrial production and mitigate over-exploitation of natural sources. High-value pharmaceuticals that include alkaloids, flavonoids, terpenes, steroids, among others, are biosynthesized as a defensive strategy by plants in response to perturbations under natural environmental conditions. However, they can also be produced using plant cell, tissue, and organ culture techniques through the application of various in vitro approaches and strategies. In the past decades, efforts were on the clonal propagation, biomass and secondary metabolites production in the in vitro cultures of medicinally important plants that produce these molecules. In recent years, the effort has shifted towards optimizing culture conditions for their production through the application of cell line selection, elicitation, precursor feeding, two-phase co-culture among cell, tissue, and organ culture approaches. The efforts are made with the possibility to scale-up the production, meet pharmaceutical industry demand and conserve natural sources of the molecules. Applications of metabolic engineering and production from endophytes are also getting increasing attention but, the approaches are far from practical application in their industrial production.     

Sugar beet extract acts as a natural bio-stimulant for physio-biochemical attributes in water stressed wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.)

The present study investigated the role of sugar beet extract (SBE) as a bio-stimulant to ameliorate the adverse effects of drought on seed germination and growth of wheat (Triticum aestivum L.). Different concentrations of SBE (0, 10, 20, 30, 40 and 50%) were used for priming the wheat seeds. The experiment was conducted in laboratory (PEG-8000 was used to create water stress) as well as under natural environmental conditions (using soil with 100 and 60% field capacity). Significant ameliorating effects of seed priming with SBE were recorded on different germination attributes, i.e., time to 50% emergence (E₅₀), germination index (GI), mean emergence time (MET), germination percentage (G%), coefficient of uniformity of emergence (CUE) and germination energy (GE) under water stress. Without priming, the plants exhibited symptoms of water stress like decreased biomass, reduction in photosynthetic pigments, e.g., chlorophyll, carotenoids. Seed pre-conditioning with SBE improved the plant growth, photosynthetic pigments, antioxidants’ activities and nutrient homeostasis of plants facing water deficit and grown under well-watered conditions. The maximum increase in biomass, content of chlorophyll, carotenoids and activities of superoxide dismutase (SOD) and peroxidase (POD) was 13.4, 8.5, 11.9, 7.6, 13.6, 42.0, 19.8%, respectively, with SBE seed priming under water stress. In conclusion, SBE seed priming effectively reduced the negativities of water stress on seed germination which resulted in better plant growth in terms of enhanced biomass, photosynthetic pigments, antioxidant defense mechanism and better nutrient homeostasis. Overall, the findings suggest that seed pre-conditioning with SBE as a bio-stimulant will be helpful for better crop stand establishment under low field capacity, especially in semi-arid and arid agricultural fields.