Application of biotechnology in breeding lentil for resistance to biotic and abiotic stress

Summary Lentil is a self-pollinating diploid (2n = 14 chromosomes) annual cool season legume crop that is produced throughout the world and is highly valued as a high protein food. Several abiotic stresses are important to lentil yields world wide and include drought, heat, salt susceptibility and iron deficiency. The biotic stresses are numerous and include: susceptibility to Ascochyta blight, caused by Ascochyta lentis; Anthracnose, caused by Colletotrichum truncatum; Fusarium wilt, caused by Fusarium oxysporum; Sclerotinia white mold, caused by Sclerotinia sclerotiorum; rust, caused by Uromyces fabae; and numerous aphid transmitted viruses. Lentil is also highly susceptible to several species of Orabanche prevalent in the Mediterranean region, for which there does not appear to be much resistance in the germplasm. Plant breeders and geneticists have addressed these stresses by identifying resistant/tolerant germplasm, determining the genetics involved and the genetic map positions of the resistant genes. To this end progress has been made in mapping the lentil genome and several genetic maps are available that eventually will lead to the development of a consensus map for lentil. Marker density has been limited in the published genetic maps and there is a distinct lack of co-dominant markers that would facilitate comparisons of the available genetic maps and efficient identification of markers closely linked to genes of interest. Molecular breeding of lentil for disease resistance genes using marker assisted selection, particularly for resistance to Ascochyta blight and Anthracnose, is underway in Australia and Canada and promising results have been obtained. Comparative genomics and synteny analyses with closely related legumes promises to further advance the knowledge of the lentil genome and provide lentil breeders with additional genes and selectable markers for use in marker assisted selection. Genomic tools such as macro and micro arrays, reverse genetics and genetic transformation are emerging technologies that may eventually be available for use in lentil crop improvement.     

Biomass production and related characters in the core collection of Pisum germplasm

Retention of residue on the soil surface following harvest is an effective method of reducing soil erosion from both wind and water. The pea crop produces small amounts of residue to effectively reduce soil erosion. Severe erosion occurs in pea production areas such as the Palouse Region of the US Pacific Northwest (PNW) when low residue crops such as spring pea or lentil are followed by fall-sown wheat. The current study was conducted to determine the range of total aboveground biomass (TAB), seed yield, and straw (residue) production from the plant identification (PI) accessions that comprise the core collection of Pisum germplasm. In addition, the potential for increasing seed yield and straw production simultaneously was evaluated. Three hundred and ninety PI accessions were screened in the field in 1996 and 1997. The variation for TAB, seed yield and straw production among the PI accessions exceeded that of the controls both years. Seed yield was positively correlated with straw production (r = 0.81, p< 0.01) indicating that seed and straw production can be increased simultaneously through positive selection for both traits. Significant favorable variation is present among accessions in the USDA core collection of Pisum germplasm which could be used to increase both seed yield and total biomass production of adapted breeding lines.     

The Effects of Mx1 Locus on Immunity Tracts Components in Large White×Meishan F2 Offspring

The mouse Myxovirus resistance protein 1 (Mx1) is known to be sufficient to confer resistance to influenza viruses, and genes encoding Myxovirus resistance protein 1 (Mx1) is, therefore, an interesting candidate gene for disease resistance in farm animals. The porcine Mxl gene has already been identified and characterized based on its homology with mouse Mxl; the full-length coding region of the pig Mxl gene spans 2 545 bp (M65087) and is organized into 17 exons compared with the human ortholog mRNA. In this study, the exons 9, 10, 11 and introns 6, 9 of the porcine Mxl gene were cloned and sequenced; two SNPs were identified in exons 9, 10, 11 but none of the SNPs led to an amino acid exchange, and the other eleven variants were detected in introns 6 and 9, respectively. Differences in allele frequency among Meishan, Large White, Tibetan, Tongcheng, Huainan, and Duroc pigs were observed within intron 9, of which an A → G substitution at position 186 was detected as an Msp Ⅰ PCR-restriction fragment length polymorphism (PCR-RFLP). The association analysis using the Large White×Meishan F2 offspring suggested that the Mxl genotype was associated with variation in several immunity traits that are of interest in pig breeding. However, further investigations in more populations are needed to confirm the above concept.     

Characterization of core collection of lentil germplasm for phenology, morphology, seed and straw yields

Increasing and maintaining crop residues in predominantly cereal-based rotations of the US Pacific Northwest is critical to controlling soil erosion. The core collection of lentil (Lens culinaris Medik.) germplasm comprising 287 accessions was evaluated for variation in phenological, morphological and growth parameters including seed yields and residue amounts over a two-year period under conventional tillage and no-till conditions. The objectives of this study were (i) assess lentil genetic variation in germplasm for variation in biomass production and seed yield, (ii) assess the relationship of phenological and morphological traits with biomass and seed yield and (iii) identify high biomass producing germplasm for use as parents in the breeding program. Days to flowering and days to maturity ranged from 31 to 78 and from 71 to 106 days, respectively. Time to flowering in terms of cumulative heat units was a more efficient measurement than days to flowering. Plant height and plant canopy width had a significant association with total biomass, seed yield and residue amounts. Total biomass ranged from 788–6389 kg ha^–1 under conventional tillage, while the range under no-till conditions was 1045–6195 kg ha^–1. Most of the lines with higher biomass also produced the highest seed yields and residue amounts. Overall, only one accession produced more residue than `Laird'. In the more favorable environment of 1997, six accessions exceeded the control cultivars, `Laird' and `Indianhead', for residue amounts, and seven and twenty-four accessions exceeded control cultivars, `Pardina' and `Brewer', for seed yield. Results indicated that plant height, canopy width at maturity and seed yield explained most of the variation in biomass and residue production. Large seeded germplasm consistently had a longer reproductive growth period than small seeded accessions and had 17%, 7% and 21% more biomass, seed yield and residue, respectively. Our data indicated significant variation in lentil germplasm for biomass, seed yield and residue amounts to warrant their use in the breeding program.     

CONGENITAL DISLOCATION OF THE DEEP DIGITAL FLEXOR TENDON ASSOCIATED WITH HYPOPLASIA OF THE SUSTENTACULUM TALI IN A THOROUGHBRED COLT

An 11-month-old Thoroughbred colt was presented with a hard swelling at the proximal third of the right 4th metatarsal bone. A medial dislocation of the deep digital flexor tendon ( flexor digitorum profundus ) was also observed on the same leg. On the plantaroproximal-plantarodistal projection of the calcaneus, there was flattening and shortening of the sustentaculum tali. The smooth bony proliferation at the proximal third of the right 4th metatarus was compatible with a chronic splint bone fracture. This report describes a medial deep digital flexor dislocation associated with hypoplasia of the sustentaculum tali .     

Synovitis Subsequent to Total-Hip Arthroplasty with and Without Hydroxyapatite Coatings: A Study in Dogs

OBJECTIVE: To establish whether hydroxyapatite (HA) coatings applied to metal total-hip-replacement components increased synovitis in the coxofemoral joint of dogs. Study Design-An in vivo research trial and histologic study. ANIMALS: Twenty-three large, mixed-breed dogs. METHODS: Arthroplasty components were implanted with HA on neither stem nor cup, or with HA applied to either the femoral component or with both stem and cup having HA applied in 23 dogs. Synovial inflammation was scored 3, 6, and 12 months after surgery. Synovial membrane thickness; polyethylene particulate load; fibrin exudate; membrane edema; mitotic index; giant-cell, mononuclear histiocyte, lymphocyte, and polymorph number; and collagen degeneration were scored and statistical analyses performed. RESULTS: Operated-side synovial thickness was greatly increased relative to control. Fibrin deposition, intramembranous edema, and collagen degeneration were similar within each time interval among the various component groups. The majority of inflammatory scores among the groups were unchanged. In this 12-month study, the presence of HA coatings appear not to have a marked impact on the development of synovial inflammation. CONCLUSIONS: HA-coated components do appear to contribute somewhat to the inflammatory load within the periprosthetic environment, but this effect is not marked. CLINICAL RELEVANCE: The major advantage of HA addition to arthroplasty components is enhanced osteoconductivity around metallic surfaces. The slight increase in particulate load seen in the presence of HA coatings may contribute to the development of aseptic loosening of arthroplasty components, but is unlikely to be the major causative factor.     

Grafting of plant embryos and the use of ultrasonics

Summary 1) Some aspects of transplantation of embryo pieces of one species, into the embryo of an other one were discussed, as well as the grafting of an embryo on the endosperm of other varieties, species and even genera (vegetative hyberidization).2) Deep hereditary changes were observed, as well as an incompatibility (negative tropism) to assimilate and to fuse between the embryonic tissues of different species.3) But a positive tropism with a complete assimilation of the tissue can also be observed in all those cases, when the grafted endosperm, belonged to the same species as the one of the transplanted embryo piece.4) This method should be extended in order to produce oil seed crops and particularily soja beans, ripening in northern long day regions (change from short day to long day photoperiodism).5) Ultrasonic vibrations could be usefully applied to improve this grafting method.

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Resume 1) Quelques aspects de transplantation d'un fragment d'embryon d'une espèce, dans celui d'une autre, ont été discutés, ainsique le gréffage d'un embryon, sur les réserves nutritives (endosperme) d'autres variétés, espèces et mêmes genres (hybridation végétative).2) Des changements héréditaires profonds ont pu être observés, ainsiqu'une incompatibilité (tropisme négative) de s'assimiler et de fusionner entre les tissus embryonaires de différentes espèces.3) Quand-même un tropisme positif, accompagné d'une assimilation complète des tissues, put être observé dans tous ces cas, ou la réserve nutritive (endosperme) appartenait à la même espèce que le fragment transplanté.4) Cette méthode devrait être approfondie pour pouvoir produire des plantes oléagineuses, en particulier du soja, qui puisse mûrir dans des régions nordiques aux jours longs, pendant la saison de croissance (changement d'un phototropisme de jours courts à celui de jours longs).5) Un traitement aux Ultra-Sons pourrait être avantageusement appliqué afin d'améliorer cette méthode.

     

The Effect of Occlusive and Semi-Occlusive Dressings on the Healing of Acute Full-Thickness Skin Wounds on the Forelimbs of Dogs

This project compared the effects of hydrocolloid (HC) and hydrogel (HG) occlusive dressings and a polyethylene (PE) semi-occlusive dressing on the healing of acute full-thickness skin wounds on the forelimbs of 10 dogs. All treatments resulted in a similar degree of healing at postoperative days 4 and 7. No significant differences existed in the number of wounds that were more than 90% healed at postoperative day 28 between the group treated with the HG dressing and the group treated with the PE dressing. There were significantly fewer wounds more than 90% healed at postoperative day 28 in the group treated with the HC dressing. Wounds under the HG dressing had the largest mean percentage of contraction at postoperative days 21 and 28. Wounds under the HG dressing also had the largest contraction/re-epithelialization ratio (postoperative days 21 and 28) compared with wounds under the PE and HC dressings. Wounds under the PE dressing had a significantly higher mean percentage of re-epithelialization than wounds under both occlusive dressings on postoperative days 14, 21, and 28. Wounds under the two occlusive dressings had exuberant granulation tissue present more often than wounds under the PE dressing. The two occlusive dressings had significantly higher bacterial counts on wounds compared with wounds under the PE dressing; analysis of variance (ANOVA), P = .0008. Wounds under the HC dressing showed the poorest healing in all parameters.     

Evaluation of auditory evoked potentials to predict depth of anaesthesia during fentanyl/fluanisone−midazolam anaesthesia in rats

Objectives To assess a method for monitoring depth of anaesthesia using components of middle latency auditory evoked potential (AEP) waveforms during anaesthesia with fentanyl/fluanisone and midazolam. Study design Prospective observational study. Animals Five female Wistar rats weighing between 210 and 250 g. Methods Implanted electrodes were used to record AEPs in animals receiving five doses of anaesthetic. Recordings were made at 5 minutes post‐injection (deep anaesthesia; no pedal withdrawal response, PWR) and then at 25 minutes (light anaesthesia; strong PWR). Responses showed five characteristic peaks occurring at 11, 14, 23, 42 and 68 ms that were measured for latency of occurrence and peak amplitude. Results Auditory evoked potential peaks P14, N23 and P42 were increased significantly in latency with successive anaesthetic injections [avg. F(1,4) = 12.53, p < 0.001; avg. F(1,4) = 10.6, p < 0.001; avg. F(1,4) = 3.9, p = 0.02, respectively]. Peak N23 showed a significant reduction in latency during the 20 minute recovery period following both the first and second anaesthetic injections (t(3) = 7.52, p = 0.005; t(4) = 5.17, p = 0.007, respectively). Peak P42 occurred significantly earlier 20 minutes following the second anaesthetic injection (t(4) = 4.75, p = 0.009). The mean overall depth of anaesthesia assessed using PWR scores was significantly correlated with the mean latency of peak N23, such that as the strength of PWR increased, N23 occurred significantly earlier (r = ?0.99, p = 0.01). The amplitude difference between peaks N23 and P42 increased after the second and third drug administrations [avg. F(1,4) = 10.65, p = 0.031 and avg. F(1,4) = 11.24, p = 0.028, respectively]. Conclusion The characteristics of these peaks, and in particular latency of peak N23, may provide a useful tool for assessing depth of anaesthesia produced by this, and possibly other anaesthetic agents.