Development of a hybrid variety system in marjoram (Origanum majorana L.)

20 accessions of marjoram (Origanum majorana L.)were characterized in two field experiments in Quedlinburg, Germany in 1997. The variation was assessed both on single plants as a base for the development of pollinator inbred lines of a prospective hybrid variety system and also on plots on a larger scale. The characterization comprised yield of leaf-flower fraction, content of essential oil and its proportion of cis-sabinene hydrate. The anther status of the regrowth after the herb harvest was used as an indicator of male fertility. The investigations revealed a high variability between and within accessions. The correlation of some traits is discussed as a clue to indirect selection. The yield rises with increasing bush diameter of the single plants and with longer vegetation periods caused by late flowering. Relations between the essential oil content and the duration of the vegetation period were not consistent and could be caused by the environment. There were no relations between the anther status and agronomical or physiological traits. This revised version was published online in July 2006 with corrections to the Cover Date.     

Time course of cannabinoid accumulation and chemotype development during the growth of <Emphasis Type="Italic">Cannabis sativa</Emphasis> L

The time course of cannabinoid accumulation in the leaves of individual plants of three Cannabis accessions was determined by gas-chromatographic analysis in greenhouse-grown plants. The total amounts and the concentration ratios of CBD, THC and CBG were determined; two accessions (an experimental hybrid, (21R × 15R) × NL, and plants from a seized seed lot) were found chemotypically uniform, with all plants belonging to chemotpe II (mixed) and I (high THC) respectively. The Carmagnola accession showed chemotypic heterogeneity, with a majority of plants belonging to chemotype III. The CBD/THC and CBG/CBD ratios were shown to be largely constant in the leaves, since 28 and until 103 days after sowing, and consistent with the ratios determined on mature inflorescences. CBD and THC maximum amounts in the leaves showed a peak in the leaves around 80 days from sowing, and were shown to be simultaneous during the growth period, irrespective of the chemotypes. Callus cultures were obtained from all the five different chemotypes (I, II, III, IV, V), and GC analyses were performed. Independently of the type and amount of cannabinoids in the mother plants, it was confirmed that callus cultures of Cannabis were not able to produce detectable amounts of any cannabinoids.     

Screening and identification of resistance to bacterial soft rot in Brassica rapa

Mist-chamber, field, and detached leaf inoculation procedures identified plants resistant to bacterial soft rot [Erwinia carotovora subsp. carotovora, (Ecc)] in Brassica rapa and related species. The mist-chamber seedling inoculation provided the best correlation of mean disease severity ratings with the field plant inoculation (r = 0.67^**) and was used to identify resistant materials. The optimum mist-chamber incubation conditions to distinguish the resistance of accessions were 23 ^°C and 100% RH and were used for primary screening. A total of 752 accessions of B. rapa and related Cruciferae were screened. In general, accessions of B. oleracea were more resistant than accessions of B. rapa. Within B. rapa, subspecies pekinensis and chinensis were more susceptible than other subspecies. No completely resistant material was found. In species of B. rapa, only 7% of accessions showed some degree of resistance with plant-to-plant variation within the accessions. G30444, G30449, and AVRDC2837 were identified as the most resistant materials in B. rapa by both mist-chamber and field inoculations. The resistance was correlated between an USA isolate (Geneva-1) and two Chinese isolates (RL4-1 and RL-19) of Ecc. This revised version was published online in July 2006 with corrections to the Cover Date.     

Population genetic structure of the clonal plant Zingiber zerumbet (L.) Smith (Zingiberaceae), a wild relative of cultivated ginger, and its response to Pythium aphanidermatum

Zingiber zerumbet (L) Smith, a wild clonal species related to the cultivated ginger (Zingiber officinale Roscoe), is a potential resistance donor for soft rot disease in ginger caused by Pythium aphanidermatum (Edson) Fitzp. In this study we evaluated the genetic diversity and P. aphanidermatum resistance of 74 Z. zerumbet accessions belonging to 15 populations from eight districts in Kerala state, India. The disease index (DI) of the accessions varied from 0% to 72.24% and the accessions could be separated into six frequency classes according to their DI values. More than 65% of the accessions had a DI < 20%. Eight accessions were found to be immune to the infection. The relative frequency of resistant accessions was higher in the central and northern regions of Kerala. Amplified fragment length polymorphism (AFLP) analysis of Z. zerumbet accessions using five primer combinations yielded 215 bands in total, of which 175 (81.4%) were polymorphic. Nei’s genetic diversity (h) of 0.2738 and Shannon information index (I) of 0.4012 revealed a high genetic diversity in Z. zerumbet unexpected for a clonal species. In the UPGMA dendrogram, accessions were clustered mostly according to their geographical origin and no clear correspondence was observed between the clustering pattern of accessions and their responses to Pythium aphanidermatum. The study revealed high genetic diversity and variability for pathogen resistance among Z. zerumbet accessions and confirmed the value of Z. zerumbet as a potential donor for soft rot resistance for the genetic improvement of ginger.     

Resistance in Lycopersicon species to black leaf mold caused by Pseudocercospora Fuligena

Over 540 accessions of wild Lycopersicon species or their crosses with L. esculentum were screened for resistance in a series of trials. Forty-six accessions were selected for the final screening trial based on lower disease ratings in previous trials. Of these, L. hirsutum had the greatest number of resistant accessions, followed by L. esculentum and L. peruvianum. Twenty accessions were quantified for their levels of resistance based on leaf area infected, area under disease progress curve (AUDPC), and the degree of sporulation. There was a significant positive correlation between the AUDPC calculated from 20 accessions evaluated under growth room and field conditions. Five L. hirsutum accessions had no sporulation associated with leaf lesions, whereas L. esculentum accessions had an average of 1.6×10⁴ conidia/cm² of leaf tissue. There was significant positive correlation between the AUDPC values and the number of conidia per cm² of leaf tissue.     

Sources of resistance to diseases of sugar beet in related Beta germplasm: II. Soil-borne diseases

Between 580 and 700 accessions of related cultivated and wild species of the genus Beta were assessed for resistance to four soil-borne diseases of sugar beet: two seedling damping-off diseases caused by the fungi Aphanomyces cochlioides and Pythium ultimum and two diseases of more mature plants, Rhizoctonia root and crown rot, caused by the fungus R. solani, and Rhizomania, caused by Beet necrotic yellow vein virus (BNYVV), a furovirus transmitted by the plasmodiophorid Polymyxa betae. Analysis of resistance data (assessed on an international standardised 1–9 scale of Resistance Scores) indicated that the highest levels of resistance ({RS} 2) to A. cochlioides and P. ultimum were to be found amongst accessions of the more distantly related sections Corollinae (93% of accessions tested) and Procumbentes (10%), respectively; although useful levels could also be found in the more closely related, and sexually compatible, section Beta (1–6%). Resistance to Rhizoctonia was also found in section Beta (5–7%), depending on whether field or glasshouse tests were used, but there was little evidence of generally high levels of resistance to Rhizomania among accessions of this section. None of the accessions of sections Corollinae and Procumbentes exhibited any notable resistance to Rhizoctonia. However, all sections Procumbentes and some sections Corollinae (4%) accessions were highly resistant to Rhizomania. Individuals with high levels of resistance to Rhizomania were identified from within some section Beta and Corollinae accessions, in which there was evidence of segregation.     

Avoidance of leaf rust fungi in wild relatives of cultivated cereals

Summary Avoidance of rust fungi that was based on poor appressorium induction was previously found in Hordeum chilense. In the present study 95 accessions of Triticeae were screened for avoidance of Puccinia hordei. The percentage of appressorium formation per germinated spore ranged from 6 to 90%. On none of the 41 accessions of Aegilops, Agropyron, Elymus, Secale, Thinopyrum or Triticum studied was the rate of appressorium formation lower than 25%. Lower rates of appressorium formation were, however, found on accessions of wild barley species Hordeum brachyantherum, H. marinum, H. parodii and H. secalinum. Its implications in cereal breeding are discussed.     

Identifying lettuce species (Lactuca subsect. Lactuca, Asteraceae): A practical application of flow cytometry

The wild lettuce species L. serriola, L. saligna, and L. virosa are important genitors in lettuce (L. sativa) breeding. Identifying these wild species can be problematic because in some cases they look very similar. Flow cytometry was tested for its reliability and general applicability as a tool to distinguish them. Three series of tests were conducted: (1) Tests with three accessions of L. sativa and one accession of each of the wild species, repeated three times throughout the year. In each repeat, the mean relative DNA amount of L. serriola was significantly higher than that of L. saligna, but significantly lower than that of L. virosa. The mean relative DNA amount of L. sativa did not differ from that of L. serriola.(2) Tests with each wild species represented by 10 accessions. Significant differences between the accessions within each species demonstrated the presence of intraspecific variation. Notwithstanding this intraspecific variation, the relative DNA amounts of all accessions of L. serriola were significantly higher than that of all L. saligna accessions, and significantly lower than that of all L. virosa accessions. Therefore, all accessions could be assigned to the appropriate species on the basis of their DNA amounts. (3) Tests with single plants from 10 accessions of each of the wild species. These test revealed that individual plants of L. serriola, L. saligna, and L. virosa can be reliably identified with flow cytometry, when aL. serriola sample of established identity is used as internal reference. This revised version was published online in July 2006 with corrections to the Cover Date.     

Germplasm diversity and estimate of genetic advance of four morpho-physiological traits in a world collection of jute

Summary To improve jute productivity, locating of sources of suitable quantitative-trait genes in the recently acquired world jute collection was needed. Two hundred sixteen accessions of Corchorus capsularis and 192 accessions of C. olitorius were evaluated at a jute seed research station (87°60\s'E, 23°80\s'N) for four yield and quality linked morpho-physiological traits. The objectives were to estimate the germplasm diversity, phenotypic (PCV) and genetic (GCV) coefficients of variation, broad sense heritability (h²) and genetic advance (GA) with respect to origins and traits. Irrespective of origins all the traits showed more than 60–80% diversity in both species. Trait means of diversity were significant but not the origin means. Generally, only slightly higher PCV values than GCV were obtained for all the traits in both the species. High h² values ranging from 50% to more than 98% were also found. All the traits can be simultaneously utilized for genetic gain. Most of the C. olitorius accessions with two times higher GA estimates than those of the C. capsularis are likely to be more successful for that matter. Maximum genetic gain is expected from Mutants > China > Brazil > India > Thailand > Nepal > Burma in C. capsularis while from Thailand > Tanzania > Mutants > Kenya > Other countries in C. olitorius. C. capsularis mutants with high heterogenity for most of the traits should be useful for finding many rare alleles.     

Reactions of Hordeum spontaneum to infection with two cultures of Puccinia hordei from Israel and United States

Summary The reactions to infection with two cultures of Puccinia hordei were determined for 292 Hordeum spontaneum (syn. H. vulgare ssp. spontaneum) accessions, collected at 16 sites which encompassed the ecological range of H. spontaneum in Israel. Culture Tel-Aviv was from Israel and culture 57.19 was from the United States. Fifty-two percent of the accessions were resistant to culture Tel-Aviv and 67% were resistant to culture 57.19. Forty-three percent of the accessions were resistant to both cultures. The average infection type (IT) of accessions within sites ranged from 2.7 to 7.5 on a 0–9 rating scale. The results showed that the presence of Ornithogalum species, the alternate hosts of P. hordei, may increase the percentage of H. spontaneoum accessions resistant to P. hordei. More accessions were resistant at sites where humidity at 1400 was higher, the annual evaporation was lower, and where the glumes were shorter. Kernel weight and annual rainfall was not correlated with resistance. A lower percentage of H. spontaneum accessions were resistant to P. hordei culture Tel-Aviv from Israel than to culture 57.19 from the United States. In a previous study a lower percentage of H. spontaneum accessions also was found to be resistant to a culture of Erysiphe graminis hordei from Israel than to cultures from other countries. Previous studies also have shown that cultures of P. hordei and E. graminis hordei from Israel have many genes for virulence on barley, and that H. spontaneum accessions from Israel have many genes for resistance to these two pathogens. Previous results and the results reported in this paper support the hypothesis of coevolution of resistant host genes and virulent pathogen genes where hosts and pathogens have coexisted for many thousand years.     

Allelism of genes controlling resistance to TMV in Capsicum L.

Summary The genes for resistance to pepper strains of TMV in five C. chinense Jacq. accessions, the PI numbers 152225, 159236, 315008, 315023 and 315024, were investigated for allelism. The resistance of these five C. chinense accessions was found to be inherited monogenically and to be partially dominant. The resistance genes in these accessions appeared to be allelic and were also found to be allelic with the known alleles L¹ and L. It is proposed to use the symbol L³ for these resistance genes.     

New sources of major gene resistance in Lactuca to Bremia lactucae

Summary A total of 1789 accessions of several lettuce collections was screened to find new major gene resistance to the downy mildew fungus Bremia lactucae Regel. The accessions belonged to the species Lactuca sativa (N=1288), L. serriola (N=399), L. saligna (N=52) and L. virosa (N=50). A total of 20 races of B. lactucae were used, 14 of which were NL-races, isolated from cultivated lettuce in the Netherlands. The other six races were isolated from wild L. serriola in Czechoslovakia. The accessions were initially screened with two races: NL1 and NL3. Accessions with resistance to one or both of these races were tested with the other races. Phenotypes with new resistance were found in accessions of all four Lactuca species. Of L. sativa, four accessions were found with resistance phenotypes that could not be explained by combinations of known major genes. Many accessions of L. serriola had resistance phenotypes that indicated the presence of unknown resistance genes. All interactions between accessions of L. saligna and races of B. lactucae were incompatible in leaf disc tests, except for four accessions, which showed some sporulation with race NL6. Several accessions of L. virosa were resistant to all races used. Other accessions of L. virosa gave a race-specific interaction with B. lactucae.     

Genetic variation and phylogenetic relationships in East and South Asian melons, Cucumis melo L., based on the analysis of five isozymes

Genetic structure and phylogenetic relationships in East and South Asian melons were analyzed, based on the geographical variation of five isozymes. The analysis of Indian melon accessions showed a continuous variation in seed length, ranging from 4 to 13 mm. Most of the East Asian melons, vars. makuwa and conomon, were classified as the small seed type with seed length shorter than 9 mm. The frequency of the small seed type increased from the west to the east in India. Allelic variation was detected at a total of nine loci of five isozymes among 114 melon accessions. Gene diversity calculated for the nine loci indicated that Indian melon was rich in genetic variation, which decreased from India towards the east. Clear geographical variation was detected in two enzymes, APS and6-PGDH. Pgd-1 ¹ and Ap-3 ¹ were frequent in India and Myanmar, while most of the melons in Laos, China, Korea and Japan carried Pgd-1 ³ and Ap-3 ³, except var. inodorusin China. Among the latter two alleles, the frequency of Ap-3 ³ was more than 50% in the small seed type in north and east India, indicating that vars. makuwa and conomon were related to the small seed type in these areas. It was also suggested that the small seed type with wet tolerance originated in central India and was selected under wet condition in the east. This revised version was published online in August 2006 with corrections to the Cover Date.     

Field screening ofPisum accessions to evaluate their susceptibility to the pea weevil (Coleoptera: Bruchidae)

Summary Seventeen unreplicated field trials over nine sites and four years were used to classifyPisum germplasm (P. sativum L. &P. fulvum Sibth. & Sm) as potential sources of resistance to the pea weevil,Bruchus pisorum (L.). The emergence of adult weevils from <10% of harvested seed was used as the selection criterion to indicate possible resistance. A total of 1900Pisum accessions were assessed using the field trials and 1754 of theP. sativum accessions were eliminated. However in the 18P. fulvum accessions screened, the level of infestation by pea weevil was always below the arbitrary resistance threshold selected. This suggests thatP. fulvum accessions could be a valuable source of resistance to the pea weevil.     

Evaluation and verification of resistance to Meloidogyne hapla Chitwood in a Cannabis germplasm collection

A collection of Cannabis accessions was evaluated for resistance to the root-knot nematode Meloidogyne hapla Chitwood in a seedling test. After inoculation with a larval suspension, significant variation was found for the number of galls and egg masses on the seedling roots. These parameters were considered as estimates for nematode infection and larval multiplication, respectively. A subset of the tested accessions was grown on a naturally infested arable field to study the relation between the test results and host characteristics in the field. The ranking order of accessions for the number of galls in the seedling test agreed well with that for the number of root galls on juvenile plants in the field. Therefore the test provides a useful indication for nematode infection. The number of egg masses in the seedling test and the population density of Meloidogyne measured in post-treatment soil samples of the field trial, showed a comparable ranking of accessions. Differences between accessions in the field were however smaller and not statistically significant.     

The inheritance of chemical phenotype in <Emphasis Type="Italic">Cannabis</Emphasis><Emphasis Type="Italic">sativa</Emphasis> L. (IV): cannabinoid-free plants

A genetic factor that blocks the cannabinoid biosynthesis in Cannabis sativa has been investigated. Crosses between cannabinoid-free material and high content, pharmaceutical clones were performed. F₁s were uniform and had cannabinoid contents much lower than the mean parental value. Inbred F₂ progenies segregated into discrete groups: a cannabinoid-free chemotype, a chemotype with relatively low cannabinoid content and one with relatively high content, in a monogenic 1:2:1 ratio. In our model the cannabinoid knockout factor is indicated as a recessive allele o, situated at locus O, which segregates independently from previously presented chemotype loci. The genotype o/o underlies the cannabinoid-free chemotype, O/o is expressed as an intermediate, low content chemotype, and O/O is the genotype of the high content chemotype. The data suggests that locus O governs a reaction in the pathway towards the phenolic cannabinoid precursors. The composition of terpenoids and various other compound classes of cannabinoid-free segregants remains unaffected. Backcrossing produced cannabinoid-free homologues of pharmaceutical production clones with potential applications in pharmacological research. A new variant of the previously presented allele ‘B ₀’, that almost completely obstructs the conversion of CBG into CBD, was also selected from the source population of the cannabinoid knockout factor.     

The exploration of genetic resources of Portuguese cabbage and kale for resistance to several Brassica diseases

Summary Twenty three accessions of nine Portuguese cabbage and kale land races from different geographic origins were tested at the seedling stage for resistance to several important brassica diseases. Resistance to downy mildew (Peronospora parasitica), expressed as necrosis of the cotyledon mesophyll, was found in all the accessions. Type A resistance to cabbage yellows (Fusarium oxysporum f. sp. conglutinans race 1) was present in most of the landraces. Resistance to clubroot (Plasmodiophora brassicae race 6) was found in one accession of the Portuguese tree kale. High resistance to blackleg (Leptosphaeria maculans) and white rust (Albuco candida) was not detected, although several accessions showed 20 to 30% of plants with intermediate expression of resistance. All Portuguese cole accessions were susceptible to blackrot (Xanthomonas campestris pv. campestris).     

The myrobalan (<Emphasis Type="Italic">Prunus cerasifera</Emphasis> L.): a useful diploid model for studying the molecular genetics of self-incompatibility in plums

A series of PCR methods were used to detect S-RNase alleles and SFB alleles and to determine S-genotypes in 25 accessions of myrobalan (Prunus cerasifera L.). Firstly, primers flanking the polymorphic second intron were used to identify S-RNases in agarose gels. These primers amplified one or two bands per accession in 25 accessions. Then consensus primers were designed for amplifying the polymorphic first intron, unique to Prunus S-RNases, for automated fluorescent detection. Each accession produced one or two peaks. New primers were then developed to amplify the intron in the SFB gene, for detection by fluorescence. Cross-referencing PCR bands and peaks indicated 15 S-alleles were present in the 25 accessions. Cloning, sequencing and comparison with published data indicated that the amplified products were S-RNase alleles. Sequence information was used to design primers specific for each S-RNase. Full and consistent S-genotypes were obtained by cross-comparing PCR data for 23 of the 25 accessions, and two accessions appeared to have a single allele. Pollen-tube microscopy indicated function of some but not all of the S-alleles sequenced.