Plant somatic mutations in nature conferring insect and herbicide resistance

Because of the role of the meristem in plant growth and reproduction, somatic mutations in plants have long been suspected of conferring herbivore and pathogen resistance on individual plants and, in the case of trees, individual branches within single plants. A few instances of resistance to phytophagous insects owing to somatic mutations have been reported in the literature. More recently, a striking example has demonstrated how somatic mutations confer resistance to an herbicide on an invasive plant, Hydrilla verticillata. The array of new methods for manipulating genomes (e.g., gene‐editing) plus existing examples of somatic mutation‐associated resistance suggest that such mutations might be useful in silviculture, agriculture, and horticulture. Answering several general questions about somatic mutations in plants would facilitate such applications: Why are so few examples reported? Do other cases exist but go undetected for want of adequate attention or methods? Under what circumstances do somatic mutations enter gametophytes? © 2018 Society of Chemical Industry     

Weed control and sensitivity of oats (Avena sativa) with various doses of saflufenacil

Saflufenacil is a new herbicide being developed by BASF for broadleaved weed control in maize, soybean and other crops prior to crop emergence. Six field studies were conducted in Ontario, Canada over a three year period (2008-2010) to evaluate the potential of saflufenacil applied pre-emergence (PRE) at various doses for broadleaved weed control in oats. Saflufenacil applied PRE caused minimal visible injury at 1, 2 and 4 weeks after emergence (WAE) in oats. At 4 WAE, the dose of saflufenacil required to provide 95% control of Ambrosia artemisiifolia (common ragweed), Chenopodium album (common lambsquarters), Polygonum convolvulus (wild buckwheat), Polygonum scabrum (green smartweed) and Sinapsis arvensis (wild mustard) was 72 to >100, >100, 74, 58 and >100 g ai ha⁻¹, respectively. Generally, similar saflufenacil dose-response trends were seen at 8 WAE. The doses of saflufenacil required to provide 95% reduction in density and dry weight ranged from 95 to >100 and 42 to >100 g ai ha⁻¹ respectively for A. artemisiifolia, C. album, P. convolvulus, P. scabrum and S. arvensis. Oat yield showed no sensitivity to saflufenacil at the doses evaluated. Based on this study, saflufenacil applied PRE can be safely used in spring planted oats for the control of some troublesome annual broadleaved weeds.     

Tolerance of spring barley (Hordeum vulgare L.), oats (Avena sativa L.) and wheat (Triticum aestivum L.) to saflufenacil

Saflufenacil is a new herbicide being developed by BASF for pre-emergence application for broadleaved weed control in maize and other crops. Three field studies were conducted in Ontario, Canada over a 2-year period (2006 and 2007) to evaluate the tolerance of spring cereals (barley, oats, and wheat) to pre-emergence and post-emergence applications of saflufenacil at 50 and 100 g ai ha⁻¹. Saflufenacil pre-emergence caused minimal visible injury (1% or less) at 3, 7, 14 and 28 days after emergence and had no adverse effect on plant height or yield of barley, oats, and wheat. Saflufenacil plus the surfactant Merge (1% v/v) applied post-emergence caused as much as 76, 60, 52 and 35% visible injury in spring cereals at 3, 7, 14 and 28 DAT, respectively. Injury with saflufenacil plus Merge applied post-emergence decreased over time and was generally greater as dose increased. Saflufenacil plus Merge applied post-emergence reduced plant height by as much as 16% and reduced yield of spring barley and wheat by 24 and 13%, respectively, but had no effect on the yield of spring oats. Based on these results, saflufenacil applied pre-emergence at the proposed dose can be safely used in spring planted barley, oats and wheat; however, the post-emergence application of saflufenacil results in unacceptable injury and yield loss. These results are consistent with the proposed pre-emergence use pattern for saflufenacil.     

Pathogenicity studies with the fungi Aphanomyces invadans, Achlya bisexualis, and Phialemonium dimorphosporum: induction of skin ulcers in striped mullet

Based on isolations from naturally infected fish in Florida, we investigated the role of the fungi Aphanomyces invadans, Achlya bisexualis, and Phialemonium dimorphosporum in the etiology of ulcerative mycosis (UM) in striped mullet Mugil cephalus. We injected healthy striped mullet subcutaneously with secondary zoospores of four oomycete isolates: two concentrations (50 and 115 zoospores/mL) of SJR (an endemic isolate of Aphanomyces invadans in American shad Alosa sapidissima from the St. Johns River); two concentrations each of CAL (25 and 65 zoospores/mL) and ACH (1,400 and 2,000 zoospores/mL; endemic isolates of Aphanomyces invadans and Achlyva bisexualis, respectively, in striped mullet from the Caloosahatchee River); and two concentrations of the ascomycete culture MTZ (2,500 and 3,500 zoospores/mL; endemic isolate of P. dimorphosporum from whirligig mullet M. gyrans in the Matanzas Inlet). All fish injected with either concentration of SJR developed granulomatous ulcers after 8 d and died within 21 d. Eighty percent (8/10) of fish injected with the high dose of CAL developed ulcers after 13 d and died within 28 d, but only 30% (3/10) of fish injected with the low dose of CAL developed ulcers. Four of the ulcerated fish died within 28 d, and the remaining fish were terminated after 32 d. Fish injected with zoospores of Aphanomyces invadans developed ulcers that were grossly and histologically similar to those observed in naturally infected striped mullet with UM from several estuaries or rivers in Florida. These hemorrhagic skin ulcers were characterized by myonecrosis and the presence of mycotic granulomas. None of the fish injected with ACH, MTZ, or sterile water developed ulcers. This study fulfilled Koch's postulates and demonstrated that ulcers could be experimentally induced in striped mullet after exposure via injection to secondary zoospores of an endemic Florida strain of Aphanomyces invadans.     

Crop–weed hybrids are more frequent for the grain amaranth ‘Plainsman’ than for ‘D136-1’

White-seeded cultivated amaranth [Amaranthus caudatus L., A. cruentus L., and A. hypochondriacus L.] grain is less valuable if contaminated with brown off-type seeds from the hybrid weedy progeny of spontaneous crop–weed crossing. Crop–weed crossing frequency was estimated by using two grain amaranth cultivars. Both cultivars were planted at the same nine locations in the Midwest United States and exposed to pollen from local weedy Amaranthus species. Harvested seeds were grown in a greenhouse, and the frequency of crop–weed hybrids was determined by observing dominant weed traits in the progeny. The cultivar ‘Plainsman’ was approximately tenfold more likely to hybridize with weedy amaranths than was ‘D136-1’. These are the first amaranth cultivars to be evaluated in this way. The reduced hybridization potential of ‘D136-1’ or similar material can be exploited to reduce the occurrence of off-type seed contamination in grain amaranths.     

Genetic characterization of the first chiropteran papillomavirus, isolated from a basosquamous carcinoma in an Egyptian fruit bat: the Rousettus aegyptiacus papillomavirus type 1

The complete genomic DNA of a novel papillomavirus (PV) was isolated from a basosquamous carcinoma on the wing of an Egyptian fruit bat (Rousettus aegyptiacus). Initial short sequences of the E1 and L1 genes of this virus were retrieved by PCR with degenerate papillomavirus-specific primers, and the entire R. aegyptiacus papillomavirus type 1 (RaPV-1) DNA was then amplified by long template PCR, cloned and sequenced with a transposon insertion method. The RaPV-1 genome counts 7970 basepairs and contains the typical papillomavirus open reading frames (ORF) (E1, E2, E4, E6, E7, L1 and L2). Based on a concatenated alignment of the E1, E2, L1 and L2 open reading frames of RaPV-1 and 46 other human and animal papillomavirus type species, a neighbor-joining phylogenetic tree was constructed. This phylogenetic analysis shows that RaPV-1 has a close-to-root position in the papillomavirus evolutionary tree. Since RaPV-1 is only distantly related to other papillomaviruses (with maximally 50% nucleotide sequence identity across the L1 open reading frame), it cannot be assigned to one of the existing papillomavirus genera and therefore represents the first member of a novel, as yet unnamed, close-to-root papillomavirus genus. This is the first time a papillomavirus has been isolated and characterized from a member of the Chiroptera order.     

Diarrhea and hyperammonemia in a horse with progressive neurologic signs

A 2-year-old, Quarter Horse filly was referred to Michigan State University, Veterinary Teaching Hospital with a 2-3 day history of depression and partial anorexia progressing to severe, watery diarrhea with severe neurologic abnormalities, including repetitive muscle fasciculations, muscle stiffening, and collapse. Laboratory findings included severe polycythemia, neutropenia, metabolic acidosis, and electrolyte and fluid loss, consistent with watery diarrhea and endotoxic shock. Increased creatine kinase and aspartate transaminase activities were consistent with recent transport and the muscle abnormalities. Severe hyperammonemia (1369.0 micromol/L; control value, 15.3 micromol/L) was found, without other substantial laboratory evidence of hepatic dysfunction. The horse was euthanized because of poor prognosis and rapid clinical deterioration. Necropsy findings were unremarkable with the exception of severe diffuse colitis. Culture of colonic contents recovered >1000 colony-forming units of Clostridium perfringens. Based on these findings, marked hyperammonemia in this filly was attributed to changes in colonic flora leading to increased bacterial production of ammonia that was readily absorbed through the inflamed bowel wall, exceeding the hepatic capacity for deamination. Intestinal bacteria as a source of hyperammonemia in the absence of hepatic disease has been linked rarely to positive culture results for clostridial organisms.     

Clearing and staining small vertebrates for demonstrating ossification of the skeletal system using the KOH and glycerine clearing method

Preparing articulated skeletons of small vertebrates is extremely difficult due to the presence of vast amounts of cartilage and/or the extremely small size of skeletal components. Clearing the tissues surrounding small skeletons and staining the bones in toto reveals the skeletal system without the risk of displacing the bony components. The specimens are first fixed and bleached which renders the tissues preserved and transparent. These specimens may then be exposed to alizarin red S which stains bone a pink to red colour making them visible through the transparent soft tissues. Toluidine blue may also be used to stain cartilage a blue colour. The Spalteholz staining method is cited, though the focus will be on the KOH and glycerine clearing method for macroscopic observation.