Mutational Analysis of a Wheat O-methyltransferase Involved in Flavonoid Metabolism

Methylated flavones, and tricin in particular, have been implicated in protecting wheat plants against a variety of biotic and abiotic stresses. Methylated flavones are produced via O-methylation of the hydroxyl groups in flavones, which is catalyzed by O-methyltransferases (OMTs). To examine the role of wheat OMT2 in methylated flavone biosynthesis and facilitate interrogation of tricin functions in wheat-environment interactions, loss-of-function mutants of OMT2 homoeologs, omt-A2 and omt-B2, were identified from a tetraploid wheat Targeting Induced Local Lesions in Genomes (TILLING) mutant population and crossed to generate the omt-A2omt-B2 double mutant. Although tricin and most other soluble phenolics did not differ in leaves and glumes of TILLING control and the omt-A2, omt-B2, and omt-A2 omt-B2 mutants, chlorogenic acid was increased in glumes of omt-A2 omt-B2 relative to TILLING control, suggesting that it might serve as a substrate for OMT2. The omt2 mutant lines showed similar growth phenotypes as well as comparable lignin deposition in cell walls of stems compared to TILLING control. These results collectively suggest that OMT2 and its close homolog OMT1 may possess overlapping activities in tricin production, with OMT1 compensating for the missing OMT2 activities in the omt2 mutant lines.

Growth, Antigenicity, and Immunogenicity of SARS-CoV-2 Spike Variants Revealed by a Live rVSV-SARS-CoV-2 Virus

SARS-CoV-2 is an emerging coronavirus threatening human health and the economy worldwide. As an RNA virus, variants emerge during the pandemic and potentially influence the efficacy of the anti-viral drugs and vaccines. Eight spike variants harboring highly recurrent mutations were selected and introduced into a replication-competent recombinant VSV in place of the original G protein (rVSV-SARS-CoV-2). The resulting mutant viruses displayed similar growth curves in vitro as the wild-type virus and could be neutralized by sera from convalescent COVID-19 patients. Several variants, especially Beta strain, showed resistance to human neutralizing monoclonal antibodies targeting the receptor-binding domain (RBD). A single dose of rVSV-SARS-CoV-2 Beta variant could elicit enhanced and broad-spectrum neutralizing antibody responses in human ACE2 knock-in mice and golden Syrian hamsters, while other mutants generated antibody levels comparable to the wild-type. Therefore, our results will be of value to the development of next-generation vaccines and therapeutic antibodies.

Arabidopsis Growth-Promotion and Root Architecture Responses to the Beneficial Rhizobacterium Phyllobacterium brassicacearum Strain STM196 Are Independent of the Nitrate Assimilatory Pathway

Phyllobacterium brassicacearum STM196, a plant growth-promoting rhizobacterium isolated from roots of oilseed rape, stimulates Arabidopsis growth. We have previously shown that the NRT2.5 and NRT2.6 genes are required for this growth promotion response. Since these genes are members of the NRT2 family of nitrate transporters, the nitrogen assimilatory pathway could be involved in growth promotion by STM196. We address this hypothesis using two nitrate reductase mutants, G5 deleted in the major nitrate reductase gene NIA2 and G′4-3 altered in both NIA1 and NIA2 genes. Both mutants had a reduced growth rate and STM196 failed to increase their biomass production on a medium containing NO3− as the sole nitrogen source. However, they both displayed similar growth promotion by STM196 when grown on an NH4+ medium. STM196 was able to stimulate lateral roots development of the mutants under both nutrition conditions. Altogether, our results indicate that the nitrate assimilatory metabolism is not a primary target of STM196 interaction and is not involved in the root developmental response. The NIA1 transcript level was reduced in the shoots of nrt2.5 and nrt2.6 mutants suggesting a role for this nitrate reductase isoform independently from its role in nitrate assimilation.

Growth rate and behaviour in separated, partially separated or non-separated kids and the corresponding milk production of their mothers

We address the hypothesis that keeping kids and mothers together would have positive effects on the milk composition of the mother and the behaviour of the kids. Kids were either permanently separated (SEP), daily separated between 7.30 and 15 h (DAY-SEP) or kept with mothers 24 h/d (NON-SEP). The NON-SEP kids were only allowed to suckle one teat. All kids had similar growth rate throughout the study (lactation days 5–70). DAY-SEP kids spent 24% of their time with their mother at both ages. NON-SEP spent only 15% of the time with their mothers at 2 weeks of age and this increased to 28% at 2 months of age. NON-SEP kids showed more hiding behaviour at 2 weeks and SEP were more active alone, at both 2 weeks and 2 months, compared to the other treatments. The mean available milk yield and fat concentration were higher in DAY-SEP goats (2420 g ± 119 g and 4.9 ± 0.1%) compared with NON-SEP goats (2149 ± 79 g and 4.4 ± 0.1%). There were no differences between DAY-SEP and NON-SEP goats in total protein, lactose, or casein concentrations. Based on these data it was estimated that 7.1 kg milk was needed to produce 1 kg semi-hard cheese in DAY-SEP goats and 7.5 kg in NON-SEP goats, respectively. When comparing milk yield and composition between udder halves, the milk yield was, as expected, higher from the machine milked teat than from the suckled one in the NON-SEP goats but there was no difference between right and left udder halves in DAY-SEP goats. Milk fat concentration varied between teats at morning and afternoon milkings in NON-SEP goats, but there was no difference in milk fat between udder-halves in DAY-SEP goats. In conclusion, the kid growth rate was similar in all treatments, however, an altered behaviour was seen in permanently separated kids (SEP). The results show that it is possible to have a high milk yield and fat concentration with one kid together with the dam.

Application of Foliar Fertilizer and Planting Media On Cattleya sp. Orchid Growth

Cattleya is one of the orchid genera, it has many fans. Labellum is large and strike colors. Intergeneric crosses have been made to produce soft colored flower, such as white, pink, red, yellow and brown. Cattleya orchids are known to have large flowers, and they are commonly referred to as The Queen of Orchid. These species are cultivated by several botanists as cut flower commodities. The price of cattleya is costly, so it becomes a business opportunity that needs to be developed. This study aims to examine the types of foliar fertilizers and planting media used on the growth of cattleya orchids. It was conducted between May and August 2021 in the experimental garden of the Faculty of Agriculture, UPN Veteran Yogyakarta using a Split Plot Design with factorial. The first factor, which is foliar fertilizer, consists of quick grow, AB mix, and focus grow. The second factor of planting media includes fern roots, black moss, and kadaka roots media. The result showed that AB Mix on fern and kadaka media produced similar plant height compared to other treatments. While on kadaka media, it produced the best leaf length and width, as well as shoot length. Furthermore, the shoot emergence and the number of roots did not show a significant difference. The plants that received quick grow treatment on kadaka and black moss media had similar growth. The number of roots provided the best growth of kadaka and ferns. Further study should be conducted to test AB Mix with various concentrations and planting media.

Replacement of Astaxanthin With Lutein in Diets of Juvenile Litopenaeus vannamei: Effects on Growth Performance, Antioxidant Capacity, and Immune Response

An 8-week feeding experiment was conducted to investigate whether diet supplementation of lutein could result in similar growth performance, antioxidant capacity, and immunity of Litopenaeus vannamei when compared to dietary astaxanthin. Juvenile L. vannamei (initial body weight: 0.64 ± 0.04 g) were fed with one of five isonitrogen and isolipids diets with/without lutein or astaxanthin [control group (C); the lutein (L) groups contained 0, 62.5, 75, 87.5 ppm lutein, respectively, the astaxanthin (A) group contained 50 ppm astaxanthin]. Results showed that dietary supplementation of lutein ranging from 62.5 to 75 ppm resulted in similar growth performance (WGR, SGR, FCR, and SR) of L. vannamei compared with the A group (P > 0.05). Apart from that, no statistical difference was observed in antioxidant parameters (hemolymph T-AOC, hemolymph MDA, and RNA expression level of GSH-PX, CAT), anti-inflammatory ability (Relish, Rho, and HSP70) and apoptosis-related gene expression (Caspase3) among lutein treatments ranging from 62.5 to 87.5 ppm and the A group (P > 0.05). These results indicate that a dose of 62.5–75 ppm of lutein was suitable in the diet of L. vannamei for substituting dietary astaxanthin.

Trends in Antiretroviral Regimen Complexity Among Medicare Beneficiaries With HIV, 2014-2018

Little is known about antiretroviral therapy (ART) patterns among Medicare beneficiaries with Human Immunodeficiency Virus (HIV). ART has significant implications for spending in Medicare Part D as use of single-tablet regimens (STR) grows, generic availability remains low, and price increases for branded therapies consistently exceed inflation. The objective of this study is to detail patterns of STR utilization among Medicare beneficiaries with HIV. We conducted a retrospective trend analysis using a 5% sample of Medicare Chronic Conditions Data Warehouse, 2014-2018. We included each person-month that fee-for-service beneficiaries with HIV had Parts A, B, and D coverage. Trends in annual prevalence of STR overall, by ART class, and by age, sex, and race subgroups were estimated. The study included 9,509 beneficiaries who contributed 345,708 person-months to the analysis. The prevalence of STR increased from 21.8% (95%CI, 21.5-22.1) in 2014 to 44.6% (95%CI, 44.3-45.0) in 2018 (p <0.0001), an increase of 104.6%. Integrase strand transfer inhibitors (INSTI) saw the largest increase in utilization between 2014 (4.4% [95%CI 4.2-4.5]) and 2018 (35.1% [95%CI 34.8-35.4]) (p<0.0001), a 701.8% increase. All sociodemographic subgroups experienced similar growth in STR use between 2014 and 2018. STR and INSTI utilization increased significantly over the study period, suggesting increased ART spending under Part D. Although increasing availability of generic multi-tablet ART regimens (MTR) may offer cost-savings, further research is needed comparing generic MTR to branded STR with regards to patient preferences, adherence, healthcare resource utilization, and total costs in the growing population of Medicare beneficiaries with HIV.

Evaluation of Microalgae Culture in PA and Commercial Urea Media

This work aimed to evaluate the growth and cost cultivation of Chlorella sp microalgae in PA and commercial urea medium solutions compared to BBM standard medium. We cultivated the microalgae Chlorella sp in BBM, PA urea, and commercial urea media, evaluating their growth for 8 days. In addition, we appraised the cost of the culture media considering the quotation of the reagents and the mass used for pilot-scale cultivation (100l). It was possible to observe the similar growth of microalgae with urea PA and BBM. The use of urea PA as a culture media for microalgae has the potential to reduce the cost of the medium by 68%. Thus, the cultivation of Chlorella in urea medium represents an alternative to reduce the production costs of biomass from this microorganism.

Effect of Biochar Application Depth on a Former Mine Technosol: Impact on Metal(Loid)s and Alnus Growth

The contamination of soil by potentially toxic elements (PTEs) is a problem resulting from various anthropic activities including the exploitation of mines, which determines an accumulation of metal(loid)s in the surrounding area. It is therefore necessary to use remediation techniques to prevent the potential damage to human health and the ecosystem. One of these techniques is phytoremediation, which involves the revegetation of contaminated areas in such a way as to reduce the spread of contaminants and entry into the groundwater by stabilizing the metal(loid)s in the soil, decreasing their mobility. To increase the ability of plants to grow under the extreme conditions of contaminated soils, it is necessary to use amendments, which can also intervene directly in reducing the mobility of contaminants. In this study, an open-field mesocosm was set up using a former mining technosol contaminated mainly by As. A biochar produced from hardwood was added at two different depths to evaluate the effectiveness of these application modalities for an overall observation duration of 17 months. Iron sulphate was also applied in both non-biochar and biochar amended conditions. In addition, trees of Alnus sp. were planted to examine the effectiveness of these plants for their use in soil remediation and the effect of the treatments used. The results showed an increase in soil pH induced by the biochar, which decreased over time. During the period examined, the application of biochar in the deepest layer was able to retain As more effectively. The Alnus sp. showed similar growth rates among the various treatments, resulting from its tolerance towards arsenic.

The Naturally Evolved EPSPS From Goosegrass Confers High Glyphosate Resistance to Rice

Glyphosate-resistant crops developed by the CP4-EPSPS gene from Agrobacterium have been planted on a massive scale globally, which benefits from the high efficiency and broad spectrum of glyphosate in weed control. Some glyphosate-resistant (GR) genes from microbes have been reported, which might raise biosafety concerns. Most of them were obtained through a hygromycin-HPT transformation system. Here we reported the plant source with 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) gene from goosegrass endowed rice with high resistance to glyphosate. The integrations and inheritability of the transgenes in the rice genome were investigated within two generations. The EiEPSPS transgenic plants displayed similar growth and development to wild type under no glyphosate selection pressure but better reproductive performance under lower glyphosate selection pressure. Furthermore, we reconstructed a binary vector pCEiEPSPS and established the whole stage glyphosate selection using the vector. The Glyphosate-pCEiEPSPS selection system showed a significantly higher transformation efficiency compared with the hygromycin-HPT transformation system. Our results provided a promising alternative gene resource to the development of GR plants and also extended the plant transformation toolbox.