Sativa Cv Research Articles

Genome-wide identification and expression analysis of DREB genes in alfalfa (Medicago sativa) in response to cold stress

ABSTRACT Dehydration-responsive element-binding proteins (DREBs) belong to members of the AP2/ERF transcription factor superfamily, which has been reported to involve various abiotic-stress responses and tolerance in plants. However, research on the DREB-family is still limited in alfalfa (Medicago sativa L.), a forage legume cultivated worldwide. The recent genome-sequence release of the alfalfa cultivar “XinJiangDaYe” allowed us to identify 172 DREBs by a multi-step homolog search. The phylogenetic analysis indicated that such MsDREBs could be classified into 5 groups, namely A-1 (56 members), A-2 (39), A-3 (3), A-4 (61) and 13 (A-5 (13), thus adding substantial new members to the DREB-family in alfalfa. Furthermore, a comprehensive survey in silico of conserved motif, gene structure, molecular weight, and isoelectric point (pI) as well as gene expression was conducted. The resulting data showed that, for cold-stress response, 33 differentially expressed MsDREBs were identified with a threshold of Log2-fold > 1, and most of which were transcriptionally upregulated within 48 h during a cold treatment(s). Moreover, the expression profiling of MsDREBs from two ecotypes of alfalfa subspecies i.e. M. sativa ssp. falcata (F56, from a colder region of Central Asia) and M. sativa ssp. sativa (B47, from Near East) revealed that most of the cold-stress responsive MsDREBs exhibited a significantly lower expression in F56, leading to a proposal of the existence of a distinct mechanism(s) for cold tolerance regulated by DREB-related action, which would have been evolved in alfalfa with a genotypic specificity. Additionally, by examining the transcriptome of a freezing-tolerance species (M. sativa cv. Zhaodong), eight DREBs were found to be implicated in a long-term freezing-stress adaptation with a great potential. Taken together, the current genome-wide identification in alfalfa points to the importance of some MsDREBs in the cold-stress response, providing some promising molecular targets to be functionally characterized for the improvement of cold tolerance in crops including alfalfa.

Reference nodule transcriptomes for Melilotus officinalis and Medicago sativa cv. Algonquin.

Host/symbiont compatibility is a hallmark of the symbiotic nitrogen‐fixing interaction between rhizobia and legumes, mediated in part by plant‐produced nodule‐specific cysteine‐rich (NCR) peptides and the bacterial BacA membrane protein that can act as a NCR peptide transporter. In addition, the genetic and metabolic properties supporting symbiotic nitrogen fixation often differ between compatible partners, including those sharing a common partner, highlighting the need for multiple study systems. Here, we report high‐quality nodule transcriptome assemblies for Medicago sativa cv. Algonquin and Melilotus officinalis , two legumes able to form compatible symbioses with Sinorhizobium meliloti . The compressed M. sativa and M. officinalis assemblies consisted of 79,978 and 64,593 contigs, respectively, of which 33,341 and 28,278 were assigned putative annotations, respectively. As expected, the two transcriptomes showed broad similarity at a global level. We were particularly interested in the NCR peptide profiles of these plants, as these peptides drive bacterial differentiation during the symbiosis. A total of 412 and 308 NCR peptides were predicted from the M. sativa and M. officinalis transcriptomes, respectively, with approximately 9% of the transcriptome of both species consisting of NCR transcripts. Notably, transcripts encoding highly cationic NCR peptides (isoelectric point > 9.5), which are known to have antimicrobial properties, were ∼2‐fold more abundant in M. sativa than in M. officinalis , and ∼27‐fold more abundant when considering only NCR peptides in the six‐cysteine class. We hypothesize that the difference in abundance of highly cationic NCR peptides explains our previous observation that some rhizobial bacA alleles which can support symbiosis with M. officinalis are unable to support symbiosis with M. sativa .

Facilitation of auxin biosynthesis and metabolism by salt stress in rice plants

To investigate the effects of salt stress on the auxin in rice plants, we analyzed indole-3-acetic acid (IAA) and its metabolites in hydroponically grown rice plant seedlings (Oryza sativa cv. Nipponbare) and in the culture medium using LC-ESI/MS/MS. We found that the IAA level in the culture medium was increased 39-fold due to salt stress treatment, while those in the shoots and roots were increased by less than 2-fold. The culture medium also contained high amounts of oxidized metabolites such as OxIAA, DiOxIAA, DiOxIAA-Glu, and DiOxIAA-Asp. The total amount of IAA and its metabolites in the shoots, roots, and culture medium was significantly increased with salt treatment. A salt stress-responsive increase in the IAA metabolites was observed in 9 tested cultivars of rice. The results indicate that salt stress facilitates the metabolic turnover of IAA in rice plants and may open new insight into the role of auxin.

Thaumatin-like Protein (TLP) Genes in Garlic (Allium sativum L.): Genome-Wide Identification, Characterization, and Expression in Response to Fusarium proliferatum Infection.

Plant antifungal proteins include the pathogenesis-related (PR)-5 family of fungi- and other stress-responsive thaumatin-like proteins (TLPs). However, the information on the TLPs of garlic (Allium sativum L.), which is often infected with soil Fusarium fungi, is very limited. In the present study, we identified 32 TLP homologs in the A. sativum cv. Ershuizao genome, which may function in the defense against Fusarium attack. The promoters of A. sativum TLP (AsTLP) genes contained cis-acting elements associated with hormone signaling and response to various types of stress, including those caused by fungal pathogens and their elicitors. The expression of AsTLP genes in Fusarium-resistant and -susceptible garlic cultivars was differently regulated by F. proliferatum infection. Thus, in the roots the mRNA levels of AsTLP7–9 and 21 genes were increased in resistant and decreased in susceptible A. sativum cultivars, suggesting the involvement of these genes in the garlic response to F. proliferatum attack. Our results provide insights into the role of TLPs in garlic and may be useful for breeding programs to increase the resistance of Allium crops to Fusarium infections.

An improved Raphanus sativus cv. WK10039 genome localizes centromeres, uncovers variation of DNA methylation and resolves arrangement of the ancestral Brassica genome blocks in radish chromosomes.

This study presents an improved genome of Raphanus sativus cv. WK10039 uncovering centromeres and differentially methylated regions of radish chromosomes. Comprehensive genome comparison of radish and diploid Brassica species of U's triangle reveals that R. sativus arose from the Brassica B genome lineage and is a sibling species of B. nigra. Radish (Raphanus sativus L.) is a key root vegetable crop closely related to the Brassica crop species of the family Brassicaceae. We reported a draft genome of R. sativus cv. WK10039 (Rs1.0), which had 54.6Mb gaps. To study the radish genome and explore previously unknown regions, we generated an improved genome assembly (Rs2.0) by long-read sequencing and high-resolution genome-wide mapping of chromatin interactions. Rs2.0 was 434.9Mb in size with 0.27Mb gaps, and the N50 scaffold length was 37.3Mb (40-fold larger assembly compared to Rs1.0). Approximately 38% of Rs2.0 was comprised of repetitive sequences, and 52,768 protein-coding genes and 4845 non-protein-coding genes were predicted and annotated. The improved contiguity and coverage of Rs2.0, along with the detection of highly methylated regions, enabled localization of centromeres where R. sativus-specific centromere-associated repeats, full-length OTA and CRM LTR-Gypsy retrotransposons, hAT-Ac, CMC-EnSpm and Helitron DNA transposons, and sequences highly homologous to B. nigra centromere-specific CENH3-associated CL sequences were enriched. Whole-genome bisulfite sequencing combined with mRNA sequencing identified differential epigenetic marks in the radish genome related to tissue development. Synteny comparison and genomic distance analysis of radish and three diploid Brassica species of U's triangle suggested that the radish genome arose from the Brassica B genome lineage through unique rearrangement of the triplicated ancestral Brassica genome after splitting of the Brassica A/C and B genomes.

Characterization and biological activities of melanin pigment from root endophytic fungus, Phoma sp. RDSE17.

Melanins are high molecular weight hydrophobic pigments which have gained popularity for their role in virulence against different pathogens. In the present study, we isolated and characterized the melanin pigment produced by a dark septate endophyte fungus Phoma sp. RDSE17, which was associated with the roots of an indigenous Oryza sativa cv. 'Chakhao amubi' in Manipur, Northeast India. The biological properties of purified melanin from the fungus were evaluated for their antioxidant, antimicrobial and anticancerous activities. The pigment was extracted from Phoma sp. by alkaline-acid hydrolysis method and confirmed as melanin through physico-chemical tests and spectral (UV, FTIR, and EPR) analysis. The analyses of the elemental composition indicated that the pigment possessed a low percentage of nitrogen (N) contents, and therefore, would not fall under DOPA class of melanin. Exposure of the fungus to melanin pathway inhibitors revealed a positive melanin inhibition by tricyclazole, but not by kojic acid. Thus, the melanin from Phoma sp. may be a member of the DHN family. Moreover, the purified melanin showed high DPPH (1, 1-Diphenyl-2-picrylhydrazyl) free radical-scavenging activity with an EC50 of 69µg/mL and inhibited human lung cancer cell (A549 cells) proliferation at 80µg/mL. The present study demonstrates that melanin from Phoma sp. RDSE17 could be employed as a potential biological (antioxidant) and antimicrobial agent for inhibiting the growth of humans and phytopathogens.

Antioxidation, Anti-Inflammation, and Regulation of SRD5A Gene Expression of Oryza sativa cv. Bue Bang 3 CMU Husk and Bran Extracts as Androgenetic Alopecia Molecular Treatment Substances.

Androgenetic alopecia (AGA), a hair loss disorder, is a genetic predisposition to sensitive androgens, inflammation, and oxidative stress. Unfortunately, current treatments with synthetic medicines contain a restricted mechanism along with side effects, whereas the bioactive constituents of plant extracts are multifunctional, with fewer side effects. The massive amounts of rice husk and bran are agricultural wastes that may cause pollution and environmental problems. Owing to these rationales, the local rice variety, Bue Bang 3 CMU (BB3CMU), which is grown in northern Thailand, was evaluated for the valuable utilization of rice by-products, husk (BB3CMU-H) and bran (BB3CMU-RB) extracts, for AGA treatment regarding antioxidant, anti-inflammatory, anti-androgenic activities, and the characterization of bioactive compounds. Our study verified that BB3CMU-H had the highest level of polyphenols, contributing to its greater antioxidant activity. Conversely, BB3CMU-RB was the predominant source of tocopherols, resulting in better anti-androgenic activities regarding the downregulation of steroid 5α-reductase genes (SRD5A). Notably, anti-inflammation via the attenuation of nitric oxide productions was observed in BB3CMU-H (0.06 ± 0.13 μM) and BB3CMU-RB (0.13 ± 0.01 μM), which were significantly comparable to diclofenac sodium salt (0.13 ± 0.19 μM). Therefore, the combination of BB3CMU-H and BB3CMU-RB could be utilized in cosmeceutical and pharmaceutical applications for AGA patients.

Nutrition by Design: Boosting Selenium Content and Fresh Matter Yields of Salad Greens With Preharvest Light Intensity and Selenium Applications.

Selenium (Se) is an essential mineral in multiple human metabolic pathways with immune modulatory effects on viral diseases including the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and HIV. Plant-based foods contain Se metabolites with unique functionalities for the human metabolism. In order to assess the value of common salad greens as Se source, we conducted a survey of lettuce commercially grown in 15 locations across the USA and Canada and found a tendency for Se to accumulate higher (up to 10 times) in lettuce grown along the Colorado river basin region, where the highest amount of annual solar radiation of the country is recorded. In the same area, we evaluated the effect of sunlight reduction on the Se content of two species of arugula [Eruca sativa (E. sativa) cv. “Astro” and Diplotaxis tenuifolia (D. tenuifolia) cv. “Sylvetta”]. A 90% light reduction during the 7 days before harvest resulted in over one-third Se decline in D. tenuifolia. The effect of light intensity on yield and Se uptake of arugula microgreens was also examined under indoor controlled conditions. This included high intensity (HI) (160 μ mol−2 s−1 for 12 h/12 h light/dark); low intensity (LI) (70 μ mol m−2 s−1 for 12 h/12 h light/dark); and HI-UVA (12 h light of 160 μ mol m−2 s−1, 2 h UVA of 40 μ mol m−2 s−1, and 10 h dark) treatments in a factorial design with 0, 1, 5, and 10 ppm Se in the growing medium. HI and HI-UVA produced D. tenuifolia plants with 25–100% higher Se content than LI, particularly with the two higher Se doses. The addition of Se produced a marked increase in fresh matter (>35% in E. sativa and >45% in D. tenuifolia). This study (i) identifies evidence to suggest the revision of food composition databases to account for large Se variability, (ii) demonstrates the potential of introducing preharvest Se to optimize microgreen yields, and (iii) provides the controlled environment industry with key information to deliver salad greens with targeted Se contents.

Variation in flowering time and flowering date stability within a cultivar of French serradella

Context Opportunity exists to expand serradella (Ornithopus spp.) use onto heavier, duplex soil types in permanent pasture environments of south-eastern Australia. This requires cultivars with appropriate flowering times and flowering-date stability (i.e. flowering at the same date in spring regardless of timing of the autumn break). Aim This work examined evidence of variation in flowering date and flowering-date stability in the NSW southern tablelands for a widely-used French serradella (O. sativus) cv. Margurita. Methods Seed (sourced from a commercial supplier) was sown 21 March 2019 (Canberra, ACT) and 231 individual plants were monitored for time to first flower. A subset of plants exhibiting ‘early’ or ‘late’ flowering were identified and their seeds collected. In 2020, seed from ~15 plants from each selection was sown in Canberra in late March and early May to represent an ‘early’ and a ‘later’ break of season (n = 3). Key results In the early-sown treatment, ‘early-flowering’ selections typically reached the median date of first flower (50%-flowering) from mid- to late August, while ‘late-flowering’ selections reached 50%-flowering early- to mid-September. When sown later, the ‘early-flowering’ selections began flowering from mid- to late September, while the ‘late-flowering’ selections flowered mid-September to early October. The ‘early’ selections exhibited greater flowering-date instability than ‘late’ selections and flowered particularly early when sown early. This indicated diversity within cv. Margurita for flowering-time control (e.g. vernalisation and/or photoperiod requirements). Implications Evaluating cultivars of serradellas for flowering date and flowering date stability in their target environment(s) is essential to ensure cultivars are suitably adapted to these environments.

Differences in biochemical, physiological and molecular response mechanisms of rice, weedy rice and barnyardgrass subjected to drought

ABSTRACT The drought stress tolerance may differ among species and cultivars. The drought effect on plants depends on the impact on the plant’s physiological, biochemical and molecular processes, as well as on its ability to adapt under these conditions. This study aimed to evaluate the physiological and biochemical responses, and determine the expression of the genes OsAPX2, OsHSP24.15, OsHSP71.10 and OsHSP85.88 under drought conditions in rice, weedy rice and barnyardgrass. A greenhouse experiment was performed in a complete randomized design, with four replications and two factors: water conditions (well-watered and water deficit) and plant species [rice (Oryza sativa cv. Puitá), weedy rice (Oryza spp.) and barnyardgrass (Echinochloa spp.)]. Under drought conditions, the rice and weedy rice plants showed more cellular damage than the barnyardgrass, and the three species showed a reduced photosynthetic rate. C3 plants (rice and weedy rice) increased the damage to lipids and proteins at 5 days of drought. However, for C4 plants (barnyardgrass), the drought conditions did not affect the biochemical parameters. The expression of the OsHSP85.88 gene increased in the three plants exposed to water deficit.

Tropical alfalfa (Medicago sativa cv. Kacang Ratu BW) supplementation for reducing cholesterol and improving quality of carcass and meat of hybrid duck

Context Duck meat is a good source of protein (21.4%) but it is high in cholesterol, with a standard range of 125–200 mg/dL. Alfalfa is a legume that contains saponins, which have been shown to reduce the cholesterol content of meat. Aim We aimed to determine the effect of alfalfa on the weight of the internal organs and carcass, the physicochemical quality of meat, and the cholesterol of hybrid ducks in Indonesia. Methods Ninety 1-day-old Indonesian hybrid ducks were fed basal rations supplemented with 0% (control), 3%, or 6% fresh alfalfa (n = 5 per group), calculated on a dry matter basis. Five replicates were made for each dietary treatment. Dietary treatments were administered to the ducks from 8 to 35 days of age. The parameters measured were the internal organ weight, carcass quality (live weight, carcass weight, carcass percentage, and abdominal fat), physicochemical quality of meat (water content, crude protein, crude fat, pH, water holding capacity [WHC], cooking loss, and tenderness), and the cholesterol content of the liver, breast meat, thigh meat, and blood (including the high-density lipoprotein/HDL and low-density lipoprotein/LDL). Analysis of variance and Duncan’s multiple range tests determined the significance of results. Key results Supplementation of rations with up to 6% of alfalfa did not significantly affect the weight of the internal organs, but reduced the cholesterol content in the liver, meat, and blood. In addition, the physicochemical parameter of alfalfa supplementation in the rations, up to a level of 6%, had no effect on carcass quality, moisture content, crude protein or pH of hybrid duck meat. Moreover, supplementation with 6% of alfalfa significantly reduced the LDL levels while increasing the HDL and affecting crude fat, WHC, cooking loss, and tenderness of hybrid ducks. Conclusions Supplementation with up to 6% of alfalfa did not affect the weight of key internal digestive organs, but reduced the cholesterol content, and increased the HDL levels of blood. Alfalfa supplementation affected the physicochemical quality of hybrid duck meat. However, the optimum level (production parameters observation) of alfalfa supplementation in the diet was at 3%. Implications Hybrid duck meat exhibits lower cholesterol, higher HDL, and lower LDL when the diet is supplemented with alfalfa, whereas other parameters were largely unchanged.

First Report of Bacterial Leaf Streak Disease of Rice Caused by Xanthomonas oryzae pv. oryzicola in Ivory Coast.

Bacterial Leaf Streak (BLS) of rice caused by Xanthomonas oryzae pv. oryzicola (Xoc) is considered as the third emerging infectious disease of rice in Africa. First reported in Africa in the 1980s, the disease is now present in at least eight African countries including Burundi, Burkina Faso, Kenya, Madagascar, Mali, Nigeria, Senegal and Uganda. Yield loss caused by BLS is estimated at 20 to 30% (Sileshi and Gebeyehu, 2021). To our knowledge BLS has so far not been reported in Ivory Coast. While BLS has not been described in the adjacent rice-growing countries Ghana and Liberia, Xoc strains isolated from samples collected between 2003 and 2011 in Burkina Faso and Mali have been characterized (Wonni et al., 2014). Xoc is transmitted through rice seeds which favors its spread through rice trading (Sileshi and Gebeyehu, 2021). Given the extensive rice trade between Burkina Faso, Mali and Ivory Coast, we hypothesized that BLS might also be present in this country. Field surveys were carried out across Ivory Coast in October 2018. Typical symptoms of the disease, e.g. translucent lesions in the form of yellow-brown to black streaks with sometimes visible droplets of exudates on the leaf surface, were observed in the area of Korhogo. 5cm-long leaf pieces were successively disinfected, rinsed in sterile water, and then ground using the Qiagen Tissue Lyser System (QIAGEN, Courtaboeuf, France). Leaf powder was resuspended in 1.5 ml of sterile water and incubated at room temperature for 30 minutes. Then, 10 μl of the suspension was streaked on semi-selective PSA medium and incubated at 28 ° C for 3 to 7 days. Colonies characteristic of Xoc, i.e. round, convex, mucous and straw yellow in color were purified from 6 individual samples from 2 distinct sites in Korhogo. To confirm their identity, isolated strains underwent a pathogenicity and molecular characterization test. The multiplex PCR developed for the identification of X. oryzae pathovars (Lang et al., 2010) revealed for all the isolates the characteristic PCR profile of Xoc (two amplicons of 324 and 691 base pairs). Strains of Xoc BLS256 and Xoo PXO99 were used as controls. The pathogenicity test was performed on 5 weeks-old plants of O. sativa cv. Azucena leaves by infiltration with a needleless syringe of a bacterial suspension at an optical density of 0.5. After 7 days of greenhouse incubation (27 ± 1°C with a 12-hour photoperiod), all infiltration points (2 infiltrations x 3 plants per isolate) developed water-soaked lesions identical to the one challenged with BLS256 while water-infiltrated leaves remained asymptomatic. These lesions were collected and subjected to the isolation and multiplex PCR processes described above, thus fulfilling Koch's postulate. Finally, three of the isolates were subjected to sequencing of the housekeeping gene gyrB by PCR amplification using the primers XgyrB1F and XgyrB1R (Young et al., 2008). Analysis of 780bp of the gyrB sequence of strains CI_k1-1, CI_k2-2 and CI_k3-2 revealed 100% identity with the gyrB sequence of Xoc reference strain BLS256 (Acc. No. CP003057) and 10 polymorphic nucleotides compared to the Xoo reference strain PXO99A (Acc. No. CP000967). To our knowledge, this is the first report of BLS in Ivory Coast supported by molecular characterization methods. New surveys in Ivory Coast and neighboring countries where the disease has not been reported will allow to implement collections and assess disease incidence as part of future control strategies.

Impact of elevated atmospheric CO2 on aleurone cells and starch granule morphology in domesticated and wild rices

Wild rice species represent valuable germplasm for breeding of elite genotypes with enhanced resistance to biotic and abiotic stresses. However, differences in endosperm morphology between wild and cultivated rice, and how these are affected by enhanced levels of atmospheric CO2 (eCO2) predicted for later this century, are largely unknown. Accessions of native rice species from northern Australia, Oryza australiensis and O. meridionalis, and a local cultivar, O. sativa cv. Doongara, were grown to maturity in glasshouses at ambient (aCO2, 400 ppm) and eCO2 (700 ppm). Endosperm morphology was studied from transverse sections of mature grains using scanning electron microscopy. Aleurone cell length and width were higher in eCO2 than in aCO2 in the O. meridionalis accession Howard Springs, but lower in Doongara. The average starch granule size in Doongara was greater than in the wild genotypes in both CO2 treatments. The area and length of the starch granule images were larger in eCO2 than in aCO2 in the O. meridionalis accession Cape York but smaller in Doongara. Responses of rice endosperm morphology to eCO2 were dependent on genotype. We conclude that endosperm morphology responds differentially to eCO2, both between and within rice species. Contrasts in these responses can be exploited by breeders.

Persistence of Escherichia coli in the microbiomes of red Romaine lettuce (Lactuca sativa cv. \u2018Outredgeous\u2019) and mizuna mustard (Brassica rapa var. japonica) - does seed sanitization matter?

BackgroundSeed sanitization via chemical processes removes/reduces microbes from the external surfaces of the seed and thereby could have an impact on the plants’ health or productivity. To determine the impact of seed sanitization on the plants’ microbiome and pathogen persistence, sanitized and unsanitized seeds from two leafy green crops, red Romaine lettuce (Lactuca sativa cv. ‘Outredgeous’) and mizuna mustard (Brassica rapa var. japonica) were exposed to Escherichia coli and grown in controlled environment growth chambers simulating environmental conditions aboard the International Space Station. Plants were harvested at four intervals from 7 days post-germination to maturity. The bacterial communities of leaf and root were investigated using the 16S rRNA sequencing while quantitative polymerase chain reaction (qPCR) and heterotrophic plate counts were used to reveal the persistence of E. coli.ResultE. coli was detectable for longer periods of time in plants from sanitized versus unsanitized seeds and was identified in root tissue more frequently than in leaf tissue. 16S rRNA sequencing showed dynamic changes in the abundance of members of the phylum Proteobacteria, Firmicutes, and Bacteroidetes in leaf and root samples of both leafy crops. We observed minimal changes in the microbial diversity of lettuce or mizuna leaf tissue with time or between sanitized and unsanitized seeds. Beta-diversity showed that time had more of an influence on all samples versus the E. coli treatment.ConclusionOur results indicated that the seed surface sanitization, a current requirement for sending seeds to space, could influence the microbiome. Insight into the changes in the crop microbiomes could lead to healthier plants and safer food supplementation.

The Combined Effect of Pseudomonas stutzeri and Biochar on the Growth Dynamics and Tolerance of Lettuce Plants (Lactuca sativa) to Cadmium Stress

Agricultural activities lead to the accumulation of cadmium (Cd) in the soil. It is necessary to identify effective and economical ways to reduce the soil Cd bioavailability. To achieve this, three bacterial strains, Pseudomonas stutzeri, P. koreensis, and P. fluorescens, were tested for tolerance and biosorption of different concentrations of Cd (0, 5, 10, 15, 20, and 25 mg L−1). During the 2020 and 2021 seasons, a pot experiment was conducted using four different soil amendments (control, biochar, P. stutzeri, and a combination) under four levels of Cd (0, 40, 80, and 120 mg kg−1) and assessing the effect on growth parameters, physiological modifications, antioxidant enzymes, and Cd accumulation in lettuce plants (Lactuca sativa cv. Balady). In vitro, the results showed that P. stutzeri was the most tolerant of Cd. Our findings in pot trials showed that T4 (biochar + P. stutzeri) was a more efficient treatment in terms of the growth parameters, with 452.00 g plant−1 was recorded for fresh weight, 40.10 g plant−1 for dry weight, 18.89 cm plant−1 for plant height, 6.03 cm2 for leaf area, and 20.48 for the number of leaves plant−1, while in terms of physiological characteristics, we recorded 1.29 mg g−1 FW, 0.35 μg g−1 FW, and 3.69 μg g−1 FW for total chlorophyll, carotenoids, and total soluble sugar, respectively; this was also reflected in the number of antioxidant enzymes and intensity of soil biological activities in soil treated with 120 mg kg−1 Cd compared with the control and other treatments in the first season. A similar trend was observed in the second season. Additionally, significantly lower Cd was observed in both the root (67%) and shoots (78%). Therefore, a combined application of biochar and P. stutzeri could be used as an alternative to mitigate Cd toxicity.

INTERACTION OF FOLIAR NUTRIENTS AND HERBICIDES INTO PRODUCTION

A field trial was carried out at the fields of Baghdad University - Jadriya for the first location during the summer season of 2018, while the second location was in Mishkab Rice Research Station, Al-Najaf al-ashraf governorate during the summer season of 2019. This study was aimed investigate rice (Oryza sativa cv. Anber) response to foliar application of nutrition with the use of different herbicides. The design was RCBD wihin using split plot arrangement four replicates. The nutrition treatments; (Humic acide, Seaweed, Nano fertilizer, Normal fertilization and Without fertilizer) were in main plots. Herbicides treatments (Ronstar, Oscar, Rainbow, Super flag, weed free and weedy) occupied the sub plots.The results indicated that Ronstar herbicide was superior in giving the lowest average weeds density value 6.55 and 9.60 plant m-2 respectively, and highest average of grain yield was 2.53 and 3.42 t.h-1, respectively. The normal fertilization treatment was superior gave the highest average grain yield value 3.07 and 4.13 t h-1. As for the effects of interaction, the Ronstar combination with normal fertilization gave highest grain yield value 3.55 and 4.59 t h-1, respectively. Concluded from this trial that the Ronstar herbicide is the best among the herbicides used with different fertilization treatments by achieving the highest averages with most of the studied charactristics.

Structural analysis of regulatory stress elements in OsCIPK14 the rice homologue and functional validation in wheat

A major mechanism of stress tolerance in plants is to increase the cytosolic concentration of Ca2+ ion, a secondary messenger that mediates cellular responsiveness to the extracellular environment. The involvement of different types of Ca2+-sensing protein kinases such as CIPK in stress responses has been elucidated in different plant models. In this study, the upstream regulatory sequence of a CIPK homologue from an Egyptian rice cultivar (Oryza sativa cv Giza182) was structurally and functionally characterized and tested for its ability to drive the expression of β- glucuronidase reporter gene (uidA) in wheat. To that end, a 1275-bp DNA fragment, 49 nucleotides upstream of the putative transcription start site (TSS) of the OsCIPK14 coding sequence, referred in this study as URS-CIPK14, was cloned, sequenced and analyzed for the presence of stress-responsiveness cis-acting elements. Subsequently, the URS-CIPK14 was fused to the uidA gene with and without the Act-1 intron, cloned in pAB6 and transformed into wheat mature embryo for transient expression. Our results showed that URS-OsCIPK14 fused with the Act-1 intron was induced by NaCl and ABA, whereas URS-OsCIPK14 drove expression of uidA constitutively in the absence of Act-1 intron. Furthermore, rice wide-genome screening using URS-CIPK14 sequence obtained in this study identified another homologue on chromosome 11. Unlike URS-CIPK14, which is associated with chromosome 12, this homologue located at about 1015 nucleotides upstream from the putative TSS of OsCIPK15 coding sequence. Interestingly, the 5’-UTS in CIPK14 is longer than CIPK15 and can code for a putative short ORF that contains two Casein kinase II phosphorylation motifs (SXXD). The significance of this ORF is not known at this time and needs to be elucidated in future studies. We conclude that data from this work suggest that URS-CIPK14 can be used as a promoter candidate for high expression level under both constitutive and inducible conditions.

Spatial Characterization of Microbial Communities on Multi-Species Leafy Greens Grown Simultaneously in the Vegetable Production Systems on the International Space Station.

The establishment of steady-state continuous crop production during long-term deep space missions is critical for providing consistent nutritional and psychological benefits for the crew, potentially improving their health and performance. Three technology demonstrations were completed achieving simultaneous multi-species plant growth and the concurrent use of two Veggie units on the International Space Station (ISS). Microbiological characterization using molecular and culture-based methods was performed on leaves and roots from two harvests of three leafy greens, red romaine lettuce (Lactuca sativa cv. ‘Outredgeous’); mizuna mustard, (Brassica rapa var japonica); and green leaf lettuce, (Lactuca sativa cv. Waldmann’s) and associated rooting pillow components and Veggie chamber surfaces. Culture based enumeration and pathogen screening indicated the leafy greens were safe for consumption. Surface samples of the Veggie facility and plant pillows revealed low counts of bacteria and fungi and are commonly isolated on ISS. Community analysis was completed with 16S rRNA amplicon sequencing. Comparisons between pillow components, and plant tissue types from VEG-03D, E, and F revealed higher diversity in roots and rooting substrate than the leaves and wick. This work provides valuable information for food production-related research on the ISS and the impact of the plant microbiome on this unique closed environment.

First Report of Meloidogyne graminicola on Rice in Henan Province, China.

Rice (Oryza sativa) is an important food crop in China and root-knot nematode Meloidogyne graminicola has been one of the most important diseases on rice in recently five years (Ju et al. 2020). In August 2020, rice plants were found to be maldeveloped, yellow leaves and hooked root tips in an irrigated paddy field of Yuanyang County, Xinxiang City, Henan Province. Fifty rice plants were randomly collected and 84.0 percent plants were infected with root-knot nematodes, with root-gall index of 56.0. Then nematodes from rice roots were isolated with 100-μm and 25-μm sieves. A large number of females, some third-stage juveniles (J3s), and a small number of males of Meloidogyne spp. were found in root galls of all samples after dissected, and then were identified and measured under the microscope. In females (n = 20), the perineal pattern was dorsoventrally oval with low and round dorsal arch, and the lateral field was not obvious or absent, striae are usually smooth, with occasional short and irregular striatal fragmentation. The morphological data of females are as follows: body length (BL) = 516.9 ± 72.5 μm (424.2 to 611.6 μm), body width (BW)= 328.4 ± 80.7 μm (232.1 to 437.4 μm), stylet length = 11.2 ± 1.3 μm (7.7 to 13.9 μm), dorsal pharyngeal gland orifice to stylet base (DGO) = 3.9 ± 0.5 μm (3.2 to 4.5 μm), vulval slit length = 24.3 ± 4.6 μm (15.2 to 31.4 μm), vulval slit to anus distance = 16.2 ± 2.5 μm (10.1 to 20.2 μm). Males are long cylindrical, wormlike, with a short round tail. Morphological measurements of males (n = 20) were BL = 1,218.0 ± 150.7μm (1,085.7 to 1,692.2 μm), BW = 34.2 ± 4.6 μm (28.5 to 39.7 μm), stylet = 17.4 ± 0.7 μm (15.9 to 19.3 μm), DGO = 3.6 ± 0.7 μm (2.5 to 4.5 μm), tail = 10.8 ± 2.1 μm (8.0 to 14.8 μm), spicule = 30.3 ± 2.6 μm (24.7 to 36.3 μm). The egg masses from the females were incubated at 28℃ for 48 hours. Measurements of J2s (n = 20) were BL = 444.2 ± 37.8 μm (315.7 to 547.5 μm), BW = 21.2 ± 2.7 μm (16.7 to 26.4 μm), stylet = 14.2 ± 0.3 μm (13.6 to 14.8 μm), DGO = 3.5 ± 0.5 μm (2.7 to 4.5 μm), tail = 70.8 ± 5.1 μm (61.3 to 80.8 μm), hyaline tail length = 21.0 ± 2.5 μm (16.3 to 26.1 μm). These morphological features are consistent with the original description by Golden and Birchfield (1965). DNA of a single female from each sample was extracted for molecular identification. Primer pairs D2A/D3B (5´-ACAAGTACCGTGAGGGAAAGTTG-3´/ 5´-TCGGAAGGAACCAGCTACTA-3´) (De Ley et al. 1999) and the species-specific primers Mg-F3/Mg-R2 (5'-TTATCGCATCATTTTATTTG-3'/ 5'-CGCTTTGTTAGAAAATGACCCT-3') (Htay et al. 2016) were used to amplify D2/D3 region of 28S RNA and the internal transcribed spacer (ITS) region, respectively. The amplified sequences of D2/D3 region (GenBank MW490724, 766 bp) shared 99.9% and 99.7% homology with the sequences of M. graminicola (MN647592, MT576694) isolated from Guangxi and Anhui Province (Ju et al. 2020), respectively, while ITS region sequences (MW487239, 369 bp) shared 100% and 99.7% homology to M. graminicola isolate GXL3 (MN636702) and FQJJ01 (MT159690), respectively. In order to verify the pathogenicity of nematodes, about 300 J2s were inoculated on ten 14-week-old rice (Oryza sativa cv. Nipponbare) planted in pots with sterilized sandy soil, respcectively, and maintained in a greenhouse at 28°C/26°C with a 16h/8h light/dark photoperiod and 75% relative humidity. At 14 days post inoculation, obvious symptoms of hook galls were observed on roots in all inoculated rice plants, and females and males in the same shape as the collected samples were found in the root galls under the stereoscopic microscope. No symptoms were observed on non-inoculated rice plants. After 28 days, the growth of the inoculated rice plants was significantly worse than that of uninoculated ones, with yellow leaves and short plants. These results confirmed the pathogenicity of M. graminicola on rice and it indicated that M. graminicola was already spread from the main rice-producing areas to the wheat and rice rotation areas. To our knowledge, this is the first report of M. graminicola in the Henan Province of China.

Oat (Avena sativa) Extract against Oxidative Stress-Induced Apoptosis in Human Keratinocytes.

Oat (Avena sativa) is well known for its various health benefits. The protective effect of oat extract against oxidative stress-induced apoptosis in human keratinocytes HaCaT was determined. First, extracts of two varieties of oat, Daeyang and Choyang, were analyzed for fat-soluble antioxidants such as α-tocotrienol, γ-oryzanols, lutein and zeaxanthin using an UPLC system and for antioxidant activity using a DPPH assay. Specifically, an 80% ethanol extract of Daeyang oat (Avena sativa cv. Daeyang), which had high amounts of antioxidants and potent radical scavenging activity, was further evaluated for protective effect against oxidative stress-induced cell death, intracellular reactive oxygen species levels, the phosphorylation of DNA damage mediating genes such as H2AX, checkpoint kinase 1 and 2, and p53 and the activation of apoptotic genes such as cleaved caspase-3 and 7 and poly (ADP-ribose) polymerase in HaCaT cells. The Daeyang and Choyang oat 80% ethanol extracts had 26.9 and 24.1 mg/100 g γ-oryzanols, 7.69 and 8.38 mg/100 g α-tocotrienol, 1.25 and 0.34 mg/100 g of lutein and 1.20 and 0.17 mg/100 g of zeaxanthin, respectively. The oat 80% ethanol extract treatment (Avena sativa cv. Daeyang) had a protective effect on oxidative stress-induced cell death in HaCaT cells. In addition, the oat 80% ethanol extracts led to a significant decrease in the intracellular ROS level at a concentration of 50–200 μg/mL, the attenuation of DNA damage mediating genes and the inhibition of apoptotic caspase activities in a dose dependent manner (50–200 μg/mL). Thus, the current study indicates that an oat (Avena sativa cv. Daeyang) extract rich in antioxidants, such as polyphenols, avenanthramides, γ-oryzanols, tocotrienols and carotenoids, has a protective role against oxidative stress-induced keratinocyte injuries and that oat may a useful source for oxidative stress-associated skin damage.