The effect of pre- and post-inoculation temperature on resistance in certain cultivars of poplar to races of Melampsora larici-populina kleb.

Summary Four cultivars of Populus spp., compatible to varying degrees with four races of M. larici-populina Kleb., were raised in a controlled environment on a high (28°/20°C, day/night) and low (20°/10°C) temperature regime. Leaf discs cut from the plants were inoculated separately with four individual races of M. laricipopulina and subsequently incubated at either low (20°C) or high (25°C) temperature for 14 days when disease development on the discs was assessed using three parameters (Incubation period to flecking, uredia per leaf disc and uredospores per mm²). The degree of resistance in all cultivar/race combinations was high on cultivars cultured at a high temperature regime compared to those cultured on a low temperature regime. Analysis of variance demonstrated that the major components: pre-inoculation temperature regime, post-inoculation temperature regime, race and cultivar, and most second and third order interactions between these were highly significant (P<0.001) for most disease parameters. The variance of the temperature components and all interactions involving these were usually higher than those for the cultivar and race components and those interactions lacking temperature components.These results emphasize the importance of the temperature regime at which plants are raised and the temperature of incubation, following the inoculation in determining the relative degree of resistance of these cultivars of poplar to races of M. larici-populina. The implications of these results in the epidemiology of leaf rust and the stability of the host-parasite relationship are discussed.     

Pathogenic variation in Melampsora medusae leaf rust of poplars

Summary Nineteen isolates of Melampsora medusae (Thüm.), collected from natural stands of Populus deltoides (Bartr.) along the lower Mississipi River Valley, were tested for the occurrence of physiologic races by inoculation of nine poplar clones. Eight distinct races were identified based upon differential responses on these clones. However, most of the isolates also differed in their aggressiveness (latent period and uredial number per leaf disk), and a significant isolate × cultivar interaction was observed for both traits. Within a specific geographic location, more than one race was present. Isolates from the northernmost sampling location (37°N latitude) appeared more aggressive than those from southern latitudes (34–36°N latitude). Thus wild pathosystems may be composed of variable pathogen populations differing in virulence and the apparent stability usually observed in such systems may be due to the genetic diversity of the host population in interaction with many epidemiological and ecological factors.The investigation reported in this paper (No. 86-8-96) is in connection with a project of the Kentucky Agricultural Experiment Station and is published with approval of the Director.     

Monogenic control of resistance in the tomato to the tobacco flea beetle: Probable repellance by foliage volatiles

Summary In comparisons made under typical summer conditions in the field at Davis, California, the foliage of the anthocyanin-deficient mutant af proved to be much more susceptible to attack by Epitrix hirtipennis (Melsheimer) than that of the isogenic normal line, normal cultivars, or any other tested mutant. Since a relatively uniform level of damage was observed in leaves of various ages that contacted the soil, the interaction is probably of a preference rather than antibiotic nature. A comparison with nine other anthocyaninless mutants proved that anthocyanin deficiency is not responsible for susceptibility. The severe reduction of glandular hairs and consequent lack of foliage aroma—hitherto unsuspected pleiotropic effects of af—probably account for the high susceptibility of this mutant. The evolutionary significance of the distinct foliage aromas characteristic of each tomato species is discussed.     

Reactions of poplar clones to physiologic races of Melampsora larici-populina Kleb.

Summary Four mono-uredospore isolates of Melampsora larici-populina Kleb. produced qualitatively distinct reactions on four half-sib clones of Populus deltoides Marsh. and thus were recognisable as races. Two of the races could be distinguished equally readily by the distinctness of their reactions (three quantitative parameters) on four compatible clones of Populus spp. The pronounced race specific reactions of the clones of P. deltoides, allied to the less distinct, but nevertheless race specific reactions, of the compatible clones are consistent with an integrated rather than a disjunctive concept of resistance in Populus spp. to M. larici-populina.     

Selection for resistance to Plasmodiophora brassicae Wor. in oriental subspecies of Brassica rapa L.

Summary Selection for resistance to Plasmodiophora brassicae Wor. in oriental groups of Brassica rapa L.Two hundred and sixty-five cultivars of leafy, oriental bassicas were tested for resistance to 18 collections of Plasmodiophora brassicae, the causal agent of clubroot. The tests were conducted in the greenhouse at low and high level inoculum concentrations. Eleven cultivars of B. rapa pe-tsai, five cultivars of B. rapa pak-choy and three cultivars of B. rapa choy-sum consistently segregated for resistance at the lower concentration of inoculum (1000 spores/ml). All 265 cultivars were susceptible at the higher concentration (1 000 000 spores/ml). Three cultivars were used in pedigree and recurrent selection schemes for increased resistance. After three cycles of selfing resistant individuals, significantly more resistant S₃ lines were derived from each cultivar. Lines derived from two cultivars. Chinese White and PI 257236, continued to improve with each cycle of selection and demonstrated increased resistance to higher levels of inoculum (up to 1 000 000 spores/ml) New cultivars based on intercrosses of S₂ resistant individuals also had significantly better resistance than the original cultivar. After two cycles of selection in the third cultivar, PI 419007, resistance did not increase and its S₂ mass did not differ significantly from the original cultivar. Evidence that indicates resistance is pathotype-non-differential and offers an alternative to major gene, pathotype-differential types of resistance currently being introduced to the leafy oriental brassicas from other Brassica rapa groups.     

Evaluation of the virulence of races 1, 2 and 4 of Fusarium oxysporum f.sp. dianthi in carnation

Summary A simultaneous analysis of the virulence of races 1, 2 and 4 of Fusarium oxysporum f.sp. dianthi to a series of nine carnation cultivars revealed the presence of different interactions between races and cultivars, as well as differences in pathogenesis between race 1 on the one hand and race 2 and 4 on the other.The most common race 2 induced typical symptoms of Fusarium wilt in all susceptible cultivars. The cultivars showed considerable variation in resistance to race 2. Only Novada remained free of external symptoms throughout the experiment. In diseased plants of all cultivars studied, infected vascular tissue was white with dark brown margins, and heavy degradation of the cell walls and xylem parenchyma cells had occurred. All Dutch isolates corresponded with race 2.Race 4 induced wilt symptoms similar to those induced by race 2, and there was a similar variation in resistance to race 2 and 4 in the cultivars. On average, the race 4 isolates were less aggressive than those of race 2. Compared with race 2, there was evidence of some genotype × race interactions: Pallas proved to be considerably more susceptible, and Lena more resistant to race 4 than to race 2. The isolates of race 4 induced a nistopathology similar to that induced by race 2, but with less vascular browning.Race 1 induced atypical but severe wilt symptoms and unusual vascular discoloration in Elsy, Niky and Sam's Pride only. The vascular tissue in these cultivars turned pale brown; in spite of heavy colonization of these tissues virtually no degradation of cell walls was observed. All other cultivars tested proved virtually resistant to race 1, providing further evidence for genotype × race interactions.Within races, limited but statistically significant genotype × isolate interactions were found as well, in particular within race 4. These are tentatively attributed to independent variation of two (or more) resistance components.     

Isolation of wheat mutants with increased resistance to powdery mildew from small induced variant populations

Small variant wheat populations created by induced mutagenesis (n = 69) or adventitious regeneration (n = 66) were intensively screened for an altered response (compared to the parent variety ‘Guardian’) to the causal pathogen of powdery mildew in wheat, Blumeria graminis f. sp. tritici. Intensive field screening following natural infection of replicated plots of wheat lines over two years revealed a total of 13 mutants exhibiting significantly greater resistance than ‘Guardian’: eight from induced mutagenesis (11.6%) of the M2 population and five from adventitious regeneration (7.6%). Complete resistance was identified in two lines, (one (M66) developed following induced mutagenesis, and the other (SC240) by adventitious regeneration). The complete resistance in the induced mutant was stable over two generations and was associated with a high frequency of leaf flecking, and consequently a low grain yield. Resistance in SC240 proved to be unstable; SC240 exhibited complete resistance to powdery mildew in the SC₂ and SC₃ generations, but only 20% of the SC₄ plants were completely resistant, while the remainder were indistinguishable in mildew response to ‘Guardian’. The mildew response of all the SC₅ generation of SC240 was not significantly different from ‘Guardian’. Yield analysis of the thirteen mutants with increased resistance in the presence of powdery mildew indicated that eleven exhibitedgrain yields at least as high as that of ‘Guardian’, while the mutant M19 exhibited a yield significantly higher than that of ‘Guardian’. This revised version was published online in July 2006 with corrections to the Cover Date.     

Identification and polymorphism of cultivars and wild ecotypes of safflower based on isozyme patterns

Summary Acid phosphatase and cathodal peroxidase isozyme patterns were used to identify nine introduced and five local cultivars of common safflower (Carthamus tinctorius L.) and seven ecotypes of wild safflower (C. oxyacantha Bieb.). Nineteen and nine bands, respectively, were readily observed for the phosphatase and peroxidase systems in the extracts from seedling shoots. Results indicated that both systems could be used jointly for complete identification of all the 21 seed sources used.Band frequencies and polymorphic indices (PI) were determined from acid phosphatase patterns using at least 92 individual seedlings of five introduced and five local cultivars and three wild ecotypes. Band frequencies ranged from complete absence in some to total presence in other populations. The mean PI for the introduced, local and wild safflowers were found to be 0.069, 0.052 and 0.058, respectively.The banding patterns of either enzyme system were similar in some instances for the cultivated and wild safflowers and there was as much variation between two cultivars as between a cultivar and a wild ecotype, indicating close genetic relationships between C. oxyacantha and C. tinctorius.     

A test of supposed mutants for stem rust resistance in wheat

Summary Eight stem rust (Puccinia graminis tritici Eriks. and Henn.) resistant lines (designated TICENA lines) that had been selected by Veiga et al. (1981) following gamma radiation of BH-1146 wheat (Triticum aestivum L.) were studied. Six of the lines were resistant to race 15B-1 of stem rust and susceptible to race 56, and proved to carry the gene Sr7a. TICENA 4 carries two unidentified genes, each giving resistance to one of the two races. TICENA 10 carries Sr6, Sr7a and an unidentified gene giving resistance to race 56 but not 15B-1. The results raise doubts about the supposed origin of the lines as mutants.     

Investigation of possible associations between partial fertility restoration and agronomic traits in Triticum aestivum L.

Summary Many conventional hard red spring wheat (Triticum aestivum L. em Thell) lines, including several North Dakota cultivars, carry a gene (or genes) which restore partial male fertility to male sterile plants with Triticum timopheevi Zhuk. cytoplasm. Since this gene has no fertility restoration function in T. aestivum cytoplasm, the postulation can be made that it is being retained in conventional lines because of pleiotropic effects, favorable linkages or chance. The research reported in this paper examined these possibilities. Forty F₆ lines, derived from a single F₂ plant which was heterozygous for a gene (or genes) for partial fertility restoration, were evaluated for two years in a yield trial planted at Fargo, North Dakota. The 40 lines were testcrossed to a male sterile line having T. timopheevi cytoplasm, and the mean seed set of testcrosses was used as a measure of a line's fertility restoration potential. Twenty-seven lines had the gene for partial fertility, and 13 lines apparently lacked this gene. The 40 lines differed for heading date, anther extrusion, plant height, grain yield, 200-kernel weight, test weight, and grain protein percentage. However, comparisons of lines having the restorer gene with those lacking the gene did not provide any obvious explanation for the retention of the partial fertility restorer gene in the breeding stocks of the North Dakota conventional hard red spring wheat breeding program. The possibility that the restorer gene was linked with genes for resistance to stem rust or leaf rust also was evaluated by testing lines for their reaction to several races of rust. No conclusive association was found.Contribution from the Agric. Exp. Sta., North Dakota State University, Fargo, ND 58105, Journal Article no.     

Use of molecular markers to accelerate the breeding of common bean lines resistant to rust and anthracnose

The main goal of this work was to introduce resistance genes for rust, caused by Uromyces appendiculatus, and anthracnose, caused by Colletotrichum lindemuthianum, in an adapted common bean cultivar through marker-assisted backcrossing. DNA fingerprinting was used to select plants genetically closer to the recurrent parent which were also resistant to rust and to race 89 of C. lindemuthianum. DNA samples extracted from the resistant parent (cv. Ouro Negro), the recurrent parent (cv. Rudá), and from BC₁, BC₂ and BC₃ resistant plants were amplified by the RAPD technique. The relative genetic distances in relation to the recurrent parent varied between 9 and 59% for BC₁, 7 and 33% for BC₂, and 0 and 7% for BC₃ resistant plants. After only three backcrosses, five lines resistant to rust and anthracnose with, approximately, 0% genetic distance in relation to the recurrent parent were obtained. These lines underwent field yield tests in two consecutive growing seasons and three of them presented a good yield performance, surpassing in that sense their parents and most of the reference cultivars tested.     

Production of mutants with high cold tolerance in spring canola (<Emphasis Type="Italic">Brassica napus</Emphasis>)

A major factor affecting spring canola (Brassica napus) production in Canada is killing frosts during seedling development in the spring and seed maturation in the fall. The objective of this study was to explore the possibility of producing spring canola lines with mutations that have altered biochemical pathways that increase cold tolerance. The approach was to generate UV point mutations in cultured microspores followed by chemical in vitro selection of individual mutant microspores or embryos resulting in measurable alterations to various biochemical pathways with elevated levels of key defense signaling molecules such as, salicylic acid (SA), p-Fluoro-d,l-Phenyl Alanine (FPA), and jasmonic acid (JA). In addition, since proline (Pro) is known to protect plant tissues in the cold-induced osmotic stress pathway, mutants that overproduce Pro were selected in vitro by using three Pro analogues: hydroxyproline (HP), azetidine-2-carboxylate (A2C); and, 3,4-dehydro-d,l-proline (DP). Of the 329 in vitro selected mutant embryos produced, 74 were identified with significant cold tolerance compared to their donor parents through indoor freezer tests at −6°C, and 19 had better winter field survival than winter canola checks. All chemically selected mutant doubled haploids with increased cold tolerance compared well with parent lines for all seed quality and agronomic parameters. Development of increased frost tolerant cultivars should allow for spring canola to be produced in western Canada without compromising seed quality.     

The effect of T-DNA on the agronomic performance of transgenic flax plants

Summary Registration and commercial release of crop cultivars improved using recombinant DNA technologies will require extensive field testing of the transgenic lines. In addition to testing the efficacy of the transferred gene(s), regulatory bodies and farmers need data from large scale agronomic studies to provide assurance that the foreign DNA does not adversely affect normal agronomic productivity or quality of the crop. Here, five transgenic sulfonylurea resistant flax (Linum usitatissimum) lines were included in a multiple location cultivar registration trial, evaluated and compared with four standard commercial cultivars and 24 other (conventionally produced) breeding lines. The transgenic lines did not appear to be affected by T-DNA for any quality or agronomic parameter tested, including yield.     

Controlled pollination transfer of a nuclear male-sterile gene from a diploid to a tetraploid watermelon line

Summary A recessive male-sterility (ms) gene was transferred from a diploid to a tetraploid watermelon (Citrullus lanatus (Thunb.) Matsum and Nakai) line through a series of controlled pollinations. Progeny testing and morphological characteristics confirmed the development of tetraploid lines containing the ms gene. The possible use of this gene transfer system in the improvement of tetraploid watermelon cultivars and the use of male-sterile tetraploids in the production of seedless triploid watermelons is discussed.     

Mutation genetics of salt tolerance in barley: An assessment of Golden Promise and other semi-dwarf mutants

A review of research at the Scottish Crop Research Institute (SCRI) on the effects of semi-dwarfing genes on salt tolerance in barley is given. Work began in1993 with the fortuitous and unexpected result that the cultivar ‘Golden Promise’ showed considerable tolerance to salt. Golden Promise is a gamma-ray induced semi-dwarf mutant of the cultivar ‘Maythorpe’. The parent and mutant cultivars are presumed to be isogenic, but show significant differences in their responses to salt stress. The positive and pleiotropic effects of the mutant gene, commonly known as GPert were found to be effective in a number of genetic backgrounds. Earlier, in 1991 Frackowiak showed that the GPert mutation was allelic to the ari-e mutants in barley. The ari-emutants were salt tested and found to show the same positive responses to salt stress as Golden Promise. This supported the allelism tests, and consequently the GPert symbol was changed to ari-e.GP. The semi-dwarf mutant sdw1 (also known as denso) and the erectoides semi-dwarf mutant,ert-k ³² were also tested for their effects on tolerance to salt, but did not show any positive effects. Salt tolerance was therefore not a general phenomenon of semi-dwarf stature but specific to mutations at the Ari-e locus in these lines. Genetic markers (RAPDs, AFLPs and SSRs) have been used for fingerprinting, genetic mapping, and QTL analysis. The markers have helped 1) confirm the isogenic relationship between Maythorpe and Golden Promise, 2)clarify the confusion over the pedigree of Golden Promise, and 3) genetically map the ari-e.GPlocus and examine its pleiotropic effects. This revised version was published online in August 2006 with corrections to the Cover Date.     

A mutant of a mutant of a mutant of a...: Irradiation of progressive radiation-induced mutants in a mutation-breeding programme with Chrysanthemum morifolium Ram.

Summary Radiation-induced sports in Chrysanthemum morifolium Ram. have been reported for several years. It has become an everyday practice to produce flower-colour mutants from outstanding cross-breeding products, even before they are distributed for the commercial production of cut flowers.One of the most successful and recent examples is that of cv. Horim, of which hundreds of mutants were produced by successive use of radiation-induced mutants in the mutation-breeding programme. Over about 4 years a variety of flower-colour mutants was obtained, not only largely including the outstanding characteristics of the original cultivar but sometimes even with an appreciable improvement in quality and yield. It is expected that the latter types, the Miros group, will soon completely supersede the spontaneous or radiation-induced Horim sports and mutants and take over the leading position of the Horim group in the production of all-year-round (AYR) cut flowers.     

In vitro selection of wheat callus tolerant to high levels of salt and plant regeneration

Summary Embryogenic calli isolated from immature embryos of four wheat cultivars were subjected to three in vitro selection methods for salt tolerance. The effect of NaCl on the selected and unselected cell lines has been investigated. The results indicated that the relative growth rate of callus decreased as the concentration of NaCl increased in both callus lines. The selected callus line gave a higher growth weight in the presence of NaCl in the medium and was highly significant as compared with unselected callus line across medium protocols in all wheat cultivars. The dry weight of both kinds of callus lines of all wheat cultivars increased markedly with increasing NaCl concentration in most cases. The Na⁺ and Cl^- contents of both callus lines were increased with increasing salinity levels while K⁺ content was decreased. The selected callus line of each cultivar at the same salinity level produced significant amounts of Na⁺, K⁺ and Cl^- higher than the unselected callus line in most salinity levels. However, the unselected callus lines of the cultivars Giza-157 and Sakha-90 at the same salinity level produced significant amounts of K⁺ higher than the selected callus line in most salinity levels. The proline content of both kinds of callus lines for all wheat cultivars was increased with increasing salinity level. However, the selected callus line gave a significantly higher proline content than the unselected callus line in all wheat cultivars at the same Salinity level. Results from the in vitro selection for NaCl tolerance showed that the stepwise method of increasing NaCl in the medium was more effective for plant regeneration than other methods.     

AFLP markers distinguishing an early mutant of Flame Seedless grape

Molecular markers have been frequently used to differentiate grape species and cultivars. There are fewer cases where molecular markers have been used to differentiate grape clones within a cultivar, or for the demarcation of somatic mutants from parental clones. This study reports the first successful utility of AFLPs for the differentiation of somatic mutants from their parental grapevine line, and discusses the potential for similar AFLP applications. The somatic mutant analysed demonstrates earlier budburst characteristics than the Flame Seedless line from which is arose. Analysis of 64 AFLP primer combinations in silver stained polyacrylamide produced in excess of 3000 markers in Vitis vinifera, and provided two markers which differentiated the somatic mutant, from its parental line. One marker was 440 bpin length and was produced with primer combinationEcoR1-AT and Mse1-CTT. The second marker was 340 bp in length and generated with primer combination EcoR1-TC and Mse1-CAC. This revised version was published online in July 2006 with corrections to the Cover Date.     

Use of Vaccinium octoploids to facilitate 4x-6x gene transfers

The use of an 8x Vaccinium produced through doubling a tetraploid hybrid with colchicine was studied as a means of facilitating gene exchange between V. corymbosum L. and V. ashei Reade. Analysis included meiotic observations and crossability studies of the 8x plant, as well as attempting to develop 6x breeding lines. Meiotic analysis revealed the presence of micronuclei in Telphase II products and polyspory in sporads. In the crossability studies five 4x highbush plants were pollinated with 8x o2 4x pollen. Data gathered included: % fruit set, average weight/fruit, average number of seeds/fruit, average number of seedlings/pollination, and number of 6x seedlings. Octoploid pollinations were significantly lower than 4x pollinations in all parameters. One 6x plant was produced from the 4x-8x crosses but was found to be mitotically unstable, having somatic cells with chromosome numbers ranging from 48 to 168.     

Mutation breeding of Chrysanthemum morifolium Ram. using in vivo and in vitro adventitious bud techniques

Summary During experiments, which are being carried out to study the factors which control the process of adventitious bud formation in vivo on detached leaves of Chrysanthemum morifolium Ram, adventitious shoots were produced from leaves, irradiated with 500 rad of X-rays. The most important but disadvantageous result was that the majority of the adventitious shoots proved to be of a chimeral nature and obviously developed from more than one cell.An in vitro adventitious bud technique was developed using different types of explants. Pedicel segments regenerated the highest number of adventitious shoots and, moreover, they developed faster as compared to explants of young flower heads or leaves. The mutants produced by irradiating the various explants were almost exclusively of a solid (non-chimeral) nature. In addition, histological observations suggest that single epidermal cells are involved in the initiation of the adventitious shoot apices.The optimum dose for mutant production is approximately 800 rad X-rays. Rather often, more than one phenotypically identical mutant was found, which was always derived from the same explant. They could for instance originate from a multi-apical meristem formed by a single mutated cell.