Weedy red rice (Oryza sativa) is a problematic weed in cultivated rice. About 50% of US rice is produced in Arkansas and 60% of these fields have some red rice infestation. Red rice populations are morphologically and phenologically diverse. We hypothesise that red rice in Arkansas has high genetic diversity, which underlies its wide phenotypic diversity, and that some alleles from cultivated rice have been introgressed into red rice during more than a century of coexistence. We tested 137 red rice accessions from four ecological zones in Arkansas and 36 cultivars that have been grown in Arkansas in the past century. Twenty-seven rice microsatellite primers, distributed across 12 chromosomes, were used to generate molecular markers. The overall NeiÕs genetic distance (GD) of red rice accessions was 0.70. Rice grown in the last century had an overall GD of 0.26. The awnless strawhull red rice was genetically distant from blackhull (GD = 0.55) and brownhull (GD = 0.60) red rice types. NeiÕs GD between blackhull and brownhull red rice was 0.42. Brownhull and blackhull formed one genotypic cluster, whereas the majority of strawhull red rice formed another cluster. Within blackhull red rice, the GD was 0.76, whereas for awnless strawhull it was 0.68, 0.75 for awned strawhull and 0.80 for brownhull types. The gene diversity of blackhull and strawhull correlated with zone of origin. A quarter of the red rice accessions share common alleles with cultivated rice. A diverse complex of weedy populations has evolved in a region devoid of other weedy and wild Oryza species.
The commercialization of imazethapyr-resistant (Clearfield™, CL) rice in the southern United States has raised serious concerns about gene flow to red rice, producing imazethapyr-resistant red rice populations. Our objectives were to determine the impact of planting date, CL cultivars, and red rice biotypes on outcrossing rate; and to investigate the relative contribution of flowering time of CL rice and red rice biotypes, together with air temperature and relative humidity (RH), on outcrossing rate. Field experiments were conducted at Stuttgart, Rohwer, and Kibler, AR, from 2005 to 2007, at three or four planting times from mid-April to late May. ‘CL161’ (inbred cultivar) and ‘CLXL8’ (hybrid) rice were planted in nine-row plots, with red rice planted in the middle row. Twelve red rice biotypes were used. The flowering of red rice and CL rice, air temperature, and RH were recorded. Red rice seeds were collected at maturity. To estimate outcrossing rate, resistance to imazethapyr was evaluated in subsequent years and confirmed using rice microsatellite markers. CLXL8 rice flowered 2 to 4 d earlier than CL161 rice, and flowering was completed within 1 wk in all plantings. The flowering duration of most red rice biotypes ranged from 4 to 17 d. Flowering synchrony of red rice biotypes and CL rice ranged from 0 to 100% at different plantings. In general, CLXL8 had greater flowering overlap and higher outcrossing rate with red rice than did CL161 rice. The outcrossing rate of red rice biotypes ranged from 0 to 0.21% and 0 to 1.26% with CL161 and CLXL8 rice, respectively. The outcrossing rate differed within each planting date (P < 0.05). Outcrossing was generally lower in mid-May and late May than in mid-April and late April planting times. Flowering synchrony and outcrossing rate were not correlated (r2 < 0.01). Outcrossing with CL161 was primarily influenced by red rice biotype. A minimum air temperature of > 24 C in the evening also favors outcrossing with CL161. With CLXL8 rice, outcrossing was most affected by RH. When RH was < 54%, outcrossing was less (0.12%) than when RH was ≥ 54% (0.38%). With CLXL8 rice, a minimum RH of ≥ 54%, from mid-morning to noon, increased outcrossing with red rice. To fully understand the interaction effects of these factors on outcrossing with red rice, controlled experiments are needed.
Red rice plants are vectors of gene flow back to cultivated rice and other weedy populations. The progeny of red rice hybrids from cultivated rice mother plants have higher chances of persistence than those from red rice mother plants. Gene flow mitigation strategies should consider this scenario.
Two red rice accessions from Arkansas have been found to be resistant to the labeled rate of imazethapyr, which is used to control red rice in ClearfieldTMrice. Full-length amplification of the acetolactate synthase (ALS) gene in imazethapyr-resistant red rice revealed a coding sequence of 1,935 base pairs, which is the same as that of the cultivated rice. Coding sequences were generated from four red rice accessions collected from different geographical regions in Arkansas, consisting of accessions that were either resistant or susceptible to imazethapyr. Nucleotide sequence alignments identified six base polymorphisms, three of which resulted in amino acid substitutions in theALSgene. One amino acid substitution, Gly654Glu, involves a residue required for imazethapyr binding to the ALS. The other substitution, Val669Met, implies conformational changes in the ALS structure that enhances binding of thiamine diphosphate, an ALS cofactor. These novel amino acid substitutions first reported for ALS-resistant red rice accessions support the hypothesis that ALS-resistant red rice can evolve with sustained herbicide selection pressure. Thus, it behooves growers to integrate the Cleafield rice technology with other tools to achieve a successful, long-term weed management program.
The inflammatory response involves a complex interplay of local tissue activities designed to recruit leukocytes and proteins from the blood to the infected tissue. For egg-type chickens, we established the growing feather ( GF ) as an accessible tissue test site to monitor tissue responses to injected test-material. For commercial broilers, whose health depends to a large extent on innate immune system functions, the GF test system offers an important novel window to directly assess their natural defenses. This study was conducted to adapt the GF test system for use in broilers, and use it to simultaneously examine local (GF) and systemic (blood) inflammatory responses initiated by GF pulp injection of lipopolysaccharide ( LPS ). Specifically, GF of 12 male and 12 female, 5-week-old broilers were injected with LPS (16 GF/chicken; 1 μg LPS/GF). Blood and GF were collected at 0 (before), 6, and 24 h after GF injection. GF pulp was used to determine leukocyte-infiltration and gene-expression profiles, reactive-oxygen-species generation, and superoxide dismutase ( SOD ) activity. Blood was used to determine blood cell profiles and SOD activity. A time effect ( P ≤ 0.05) was observed for most aspects examined. In GF, LPS injection resulted in heterophil and monocyte infiltration reaching maximal levels at 6 and 24 h, respectively. Reactive-oxygen-species generation, SOD activity, and mRNA levels of IL-1β, IL-8, IL-6, IL-10, and cathelicidin B1 were elevated, whereas those of TNF-α, LITAF, SOD1, and SOD2 decreased after LPS injection. In blood, levels of heterophils and monocytes were elevated at 6 h, lymphocytes and RBC decreased at 6 h, and thrombocytes and SOD activity increased at 24 h. Assessment of LPS-induced activities at the site of inflammation (GF) provided novel and more relevant insights into temporal, qualitative, and quantitative aspects of inflammatory responses than blood. Knowledge generated from this dual-window approach may find direct application in identification of individuals with robust, balanced innate defenses and provide a platform for studying the effects of exogenous treatments (e.g., nutrients, probiotics, immunomodulators, etc.) on inflammatory responses taking place in a complex tissue.
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