Gummy stem blight resistance of cucumbers (Cucumis sativus L.)

Summary A start was made with breeding for resistance to gummy stem blight in cucumber. A method has been development for screening plants in a young stage. Using this method a distinct level of resistance was found in plants of Leningradsky, Wjarnikovsky,a P.I. entry from Birma, Rheinische Vorgebirge and a P.I.entry from Turkey. Lines developed from this material show a higher level of resistance than Dutch slicing and pickling cucumber varieties.     

Micropropagation of mother plants of lucerne (Medicago sativa L.) for somatic embryogenesis

Summary A reliable and standard method was established for micropropagation of the A70-34 selected genotype of lecerne (Medicago sativa L., genotype A70-34), aimed at reducing contamination problems, seasonal and phenological influences on regeneration and phytosanitary problems of the mother plants, while maintaining the regenerative potential for somatic embryogenesis of the plant tissues. Mother plants were routinely maintained for several subcultures and somatic embryogenesis was regularly obtained from the subcultured explants. Proliferation, rooting and embryogenetic ability of plants cultured for 30 days was greater than those cultured for 20 days. The regenerative potential of tissues from different organs and of triturated and intact whole plants was also tested. Petioles were confirmed as the best source for embryogenesis as far as efficiency and repeatability were concerned, even though regeneration from other explant types was also achieved. Production of somatic embryos through mechanical trituration of the in vitro cultured plants was obtained; the regeneration ability of the triturated plants was greater than that observed in the intact plants.     

A new method for selecting cytochimeras by the maximum number of nucleoli per cell in Allium wakegi Araki and A. fistulosum L.

Summary A new method for determining ploidy levels in cytochimeral plants was developed by examining the maximum number of nucleoli per cell. Colchicine-treated plants of Allium wakegi Araki and A. fistulosum L., which have different ploidy levels in the first (LI), the second (LII), and the third (LIII) germ layer, were used together with untreated 2-2-2 plants of the same species. Nucleoli in guard cells for LI and in mesophyll cells for LII were stained in a 50% AgNO₃ aqueous solution at 55° C for three hours under dark, humid conditions. In both species the ploidy levels of LI determined by the maximum number of nucleoli per guard cell accorded well with those determined by guard cell length. In A. fistulosum the ploidy levels of LII determined by the maximum number of nucleoli per mesophyll cell clearly agreed with those determined by pollen size. This method provided more precise and efficient identification for LI and LII ploidy than the conventional methods of using guard cell length for LI and pollen size for LII.     

Field performance of embryogenic cell suspension-derived banana plants (Musa AAA, cv. Grande naine)

Growth and yield characteristics of two different clones of banana plants (Musa AAA cv. Grande naine) originating from four months old embryogenic cell suspensions were studied. These characteristics were compared with those plants produced by the conventional in vitro budding multiplication method. Two types of variants were observed during the acclimatization phase among 500 embryogenic cell suspension derived plants. The first type related to banana plants with `variegated or deformed leaves' were also observed in in vitro budding derived plants. The second type concerned `fasciated-leafed' plants. During the field growth, these two variant types produced plants morphologically similar to the other plants. Thus, none of the cell suspension derived plants exhibited off-type traits in the field. A Fisher block model was used to compare the field performances of the two clones produced through the two in vitro propagation techniques. The analysis of variance showed that there were no significant differences between the plants produced by either micropropagation techniques for the plant height and circumference, the length of the reference leaf, the number of nodal clusters of the inflorescence and of fruits, the bunch weight, the period of time between planting and flowering, and between planting and harvesting. This study showed that banana plants with an agronomical behaviour similar to those produced by the conventional in vitro budding method could be regenerated from embryogenic cell suspension. This revised version was published online in July 2006 with corrections to the Cover Date.     

Indicators of resistance of sunflower plant to basal stalk rot (Sclerotinia sclerotiorum): Symptomatological,biochemical, anatomical,and morphological characters of the host

Summary The object of this work was a study of the relationship between the field reactions of different sunflower genotypes to basal stalk rot (in terms of severe (dead plants) and incipient wilting, and lesion length) and some biochemical (phenol concentration), morphological (plant height, and stem and flower-bud diameters) and anatomical (xylem and cortical indexes) characters of the host. Plants from 8 inbred lines at closed flower-bud stage were artificially inoculated with mycelium at the base of the stem. The percentage of dead plants for each inbred line and the lesion length and wilting range for individual plants after 7 days were recorded.A positive and highly significant correlation coefficient between the percentage of dead plants and lesion length was found for the three years of the study (r=0.83; P<0.01). A highly significant association between lesion length and wilting range for individual plants was always found (P=0.00). Postinfectional phenol content exhibited a strong negative correlation with lesion length and the percentage of dead plants in all the experiments (P=0.05). Association between postinfectional phenol content and wilting range for individual plants was significant for all the years studied (P<0.05). No correlation between phenol levels in healthy plants of the different sunflower genotypes and their susceptibility was found. Morphological characters positively correlated with lesion length but only plant height exhibited significant values for the three years. Associations between wilting range and morphological characters for individual plants were significant for one of the two years analyzed (P<0.05). Xylem index showed a negative correlation with lesion length which was significant one of the two years studied.The lesion length measure seems to be a simple and direct method for resistance screening before the flowering period. Although strong relations with postinfectional phenol levels were found, their determinations would be too much time consuming and not completely reliable. The relationships between other characters measured and disease resistance would indicate that physiological mechanisms could be related to resistance.     

Gene dispersal from genetically modified oil seed rape in the field

Summary A field trial was carried out with oil seed rape plants that had been genetically modified to contain genes coding for marker characters (-glucuronidase, kanamycin resistance and asulam resistance). The aim of the experiment was to examine a method of studying gene dispersal in an agricultural environment. The central area of the field plot comprised 150 genetically modified plants and 450 plants not expressing the marker genes. These were surrounded by additional non-expressing plants. The plants were allowed to freely cross pollinate and a sample of the resultant progeny from non-expressing plants were screened for expression of the marker characters and for the presence of the introduced genes (by PCR). Limited gene dispersal was detected and the frequency of modified seedlings amongst the progeny of a plant appeared to have been strongly influenced by the genotype of the immediately adjacent plants.     

Characterization of CMS and maintainer lines in indica rice (Oryza sativa L.) based on RAPD marker analysis

Total DNA from WA type CMS lines: Zhenshan 97 A, Longtepu A and theirmaintainers Zhenshan 97 B, Longtepu B wasextracted by CTAB method. One hundredprimers were used for screening RAPDmarkers to distinguish CMS line (A) andmaintainer (B) plants at seedling stage.Results showed that under the conditions of37 ^°C annealing temperature and1.5 mM MgCl₂ concentration, in Zhenshan97 A, Longtepu A there was a 1600 bp DNAfragment in product amplified by primerOPA12, while in Zhenshan 97 B, and LongtepuB no 1600 bp fragment was found. The 1600 bpfragment was also found in DA type CMS lineXieqingzao A, but was absent in XieqingzaoB. Also in the restorer line, Minghui 63the 1600 bp fragment was absent. In F₁and F₂ generation of Zhenshan97A/Minghui 63, all plants investigated hadthe 1600 bp fragment. When mitochondrial DNA(mtDNA) was isolated from the three CMS (A)and their B lines and amplified by OPA12,results showed that the 1600 fragment wasfound in all the three A lines and wasabsent in the three B lines. In DA typeXieqingzao A, two other fragments (700 bp,1000 bp) were also found except the 1600 bp.These results indicate that the 1600 bpfragment derived from CMS mitochondrial DNAcan be used as a RAPD marker to distinguishA and B plants at seedling stage, and thefragments (700 bp, 1000 bp) can be used todistinguish WA and DA cytoplasms.     

RAPD variability in rice (Oryza sativa L.) plants derived from desiccation-tolerant calli

Genetic modification from selfed progenies of 18 rice (Oryza sativa L.) plants regenerated from callus tissues which survived desiccation, were investigated at the DNA level using the random amplified polymorphic DNA (RAPD) method. Twelve 10-mer random primers were used to amplify DNA of progenies from the regenerated plants, and a total of 228 PCR products and 1780 DNA fragments were obtained by primers, generating between four to thirteen major bands. The size of the amplified fragments ranged from 0.2 to 2.55 kb. The results showed that 10 out of 12 primers produced polymorphic bands, two primers (RA31 and RA185) showed no polymorphism among plants tested. A dendrogram of the genetic distance was constructed based on their polymorphism, demonstrating that somaclonal variation exists in rice plants regenerated from callus which survived the desiccation treatment. Part of this variation can be useful in rice breeding. This revised version was published online in August 2006 with corrections to the Cover Date.     

Production and performance of potato mini-tubers

Summary Production of mini-tubers as a source for seed potato was investigated by growing in soil micropropagated plants and micro-tubers produced from micropropagated plants. Cultures of several cultivars were initiated from indexed tubers and multiplied on modified MS medium. Cultures were micropropagated by using a modular system which allowed batch handling. Micropropagated plants produced mini-tubers in glasshouse after 70–115 days of growth in soil. A large proportion of the mini-tubers produced were between 9 and 15 mm diameter. Several factors, e.g., explant number, duration of in vitro culture and genotype influenced mini-tubers production. Micropropagated plants after culture of 86 days or longer produced micro-tubers ca. 2 to 10 mm diameter. Plants, which formed micro-tubers in vitro, produced less number of mini-tubers in soil. Micro-tubers produced 1 to 3 mini-tubers when grown in soil in chain-type paper pots, but produced conventional sized tubers when grown in soil under plastic polytunnel. Mini-tuber number varied widely between potato cultivars; cvs. Bintje and British Queen produced more mini-tubers than the other cultivars. Mini-tubers developed green hard skins when kept in light for 3 weeks, and could be stored in dark at 4° C upto 6 months. In a field trial, small mini-tubers ca. 5–10 mm diameter produced more but smaller tubers than mini-tubers ca. 15–20 mm diameter. The micropropagated plants and the plants grown from mini-tubers were genetically stable, and did not show any morphological aberrations except for one variegated plant among the several thousand produced. It is concluded that the production of mini-tubers by soil planting of micropropagated plants is a rapid and efficient method for producing seed potato tubers.The use of any trade mark and other commercial products mentioned in this paper does not constitute any recommendation or advertisement for such products on behalf of the Agriculture and Food Development Authority, Dublin, Ireland     

Nondestructive assessment of protein content in single seeds of rapeseed (Brassica napus L.) by near-infrared reflectance spectroscopy

The potential of near-infrared reflectance spectroscopy (NIRS) to detectwithin-plant differences for seed protein content was investigated. Fourhundred and fifty-one single seeds were scanned by NIRS using a specialadapter. After non-destructive NIRS scanning, the seeds were analysed forprotein content by the Dumas combustion method and a calibrationequation was developed. A validation set of 117 additional seeds fromthree individual plants from the cultivars Bristol, Lirajet and Maplus wasanalysed for protein content both by NIRS and combustion. The coefficientof determination between NIRS and combustion values in the validation setwas 0.94, with a standard error of performance (SEP) of 0.77% and aratio of the SEP to the standard deviation (SD) of the validation set of0.28. The coefficient of variation (CV) for seed protein content inindividual plants, as determined by the combustion method, was 11.7%for Bristol, 8.9% for Lirajet, and 9.5% for Maplus. The comparison ofsuch variation with the standard error (SE) of NIRS analysis, defined as thecombination of the SE of the combustion method and the SEP of NIRScalibration equation, revealed that the maximum explainable variance withinindividual plants that can be detected using NIRS analysis of proteincontent in single seeds was 0.86 for Bristol, 0.83 for Lirajet, and 0.85 forMaplus. These results demonstrated that NIRS is a powerful tool fornon-destructive assessment of within-plant variation for seed protein contentin rapeseed.     

Trueness-To-Type and Yield Components of the Banana Hybrid Cultivar FHIA-18 Plants Regenerated Via Somatic Embryogenesis in a Bioreactor

Summary A population of 1,500 plants of the banana hybrid ‘FHIA-18’ (AAAB), regenerated from somatic embryos, which were multiplied in bioreactors, showed similar characteristics to plants propagated from shoot tip cultures both in the acclimatization stage and in field experiments carried out in Cuba. The plants originating from somatic embryos were similar to the plants obtained from shoot tips with respect to plant height, diameter of the pseudostem and number of suckers. Both groups of plants obtained from in vitro cultures were significantly different to the plants obtained from suckers during the flowering period of the mother plants, which was shortened by two months. The greater plant height and diameter of the pseudostem in the plants coming from somatic embryos and shoot tip is due to the effect of in vitro culture, and this was observed in different banana and plantain cultivars. During the second cycle of evaluation, the plants coming from the three propagation methods studied in this work had similar growth habits without significant differences in the majority of the morphological parameters evaluated. These results confirm that the difference obtained during the first cycle between the distinct populations is attributed to temporary changes. The original characteristics of the cultivar were evident from the second cycle of culture. Only 0.13% somaclonal variant was observed in the plants coming from somatic embryogenesis. These percentages are low taking into consideration that other propagated methods accept up to 5% variants in field conditions.     

A sampling technique for assessing digestibility of Italian ryegrass populations

Summary A sampling technique is proposed for rapid digestibility screening of herbage grass ecotypes. Its application in the characterisation of genetic resources is discussed. The technique aims to standardize maturity and circumvent the problems of variation in stem:leaf ratio and the use of single plants versus swards.A experiment comparing the method with a standard technique using eleven ecotypes plus two controls of Lolium multiflorum is described.     

New interspecific hybrids in the genus Agropyron

Summary Fifteen species from the genus Agropyron were crossed together. Fourteen of the crosses did not produce fruits. Twenty crosses produced varying numbers of caryopses. Of the fruits produced, 22% contained no embryos. For a variety of reasons it was possible to obtain only 2 plants from the remaining 140 embryos. The crosses which yielded viable plants were between A. trachycaulum cv. Primar (2n=28) and A. intermedium cv. Chief (2n=42) as well as between A. trachycaulum from Lethbridge (2n=28) and A. desertorum cv. Nordan (2n=28). The somatic chromosome numbers for the hybrids are 37 and 28. respectively. Studies of vegetative plant character are presented.     

Comparison of inoculation methods with Didymella bryoniae on Cucumis sativus

Summary A good correlation between foliage attack of young and mature plants was observed. No evidence could be found that plants showing some resistance to plant infection produce fruits less susceptible to internal fruit rot. The extent of external fruit rot depended on the extent of wounding and on the thickness of the peel.     

Selection of hybrid tea-rose seedlings for cut flower yield

Summary In a three year's trial in the greenhouse, flower yields of seedling Hybrid Tea-roses both on their own roots and on a rootstock were recorded. Seedlings on their own roots yield similarly to plants on a rootstock. Highly significant correlations were found annually between the two categories. A selection method for potential yield on the basis of the first year's yield of seedlings on their own roots, is described. Particularly in combination with selection for flower quality this method saves time, labour and space.     

Development of black rot resistant interspecific hybrids between Brassica oleracea L. cultivars and Brassica accession A 19182, using embryo rescue

Black rot is a bacterial disease of Brassica oleracea caused by Xanthomonas campestris pv. campestris. Resistance to the major black rot races 1 or 4 has been identified in related Brassica species including B. carinata and B. napus. In this study, two B. juncea accessions (A 19182 and A 19183) that are resistant to races 1 and 4 of Xcc were used as maternal and paternal parents to generate interspecific hybrids with B. oleracea cultivars. Interspecific hybrids were recovered using the embryo rescue technique and confirmed through inheritance of paternal molecular markers. Twenty-six interspecific hybrid plants were obtained between A 19182 and B. oleracea cultivars, but no interspecific hybrids were obtained using A 19183. Although interspecific hybrid plants were male sterile, they were used successfully as maternal parents to generate backcross plants using embryo rescue. All hybrid and BC₁ plants were resistant to black rot races 1 and 4.     

Production and characterization of intergeneric diploid cybrids derived from symmetric fusion between Microcitrus papuana Swingle and sour orange (Citrus aurantium)

Plants were regenerated from intergeneric somatic hybridization between embryogenic protoplasts of Microcitrus papuana Swingle and leaf-derived protoplasts of sour orange (Citrus aurantium L.) via electrofusion. The regenerated plants were morphologically similar to the leaf parent in growth vigor, leaf and branch structure. FCM analysis showed that they were diploids. Simple-sequence-repeat (SSR) and cleaved-amplified-polymorphic-sequence(CAPS) were employed for hybridity characterization. SSR banding patterns of the regenerated plants were identical to the leaf parent, sour orange, indicating that they possessed nuclear component derived from sour orange. DNA amplification with chloroplast and mitochondrial universal primers, followed by restriction endonuclease digestion, revealed polymorphism between the fusion parents. Therefore, this method was used to determine the cytoplasmic compositions of the regenerated plants. Banding patterns for all the polymorphic primer/enzyme combinations of the regenerated plants were similar to those of the embryogenic parent, M. papuana, suggesting that only the cytoplasmic components derived from the embryogenic parent were present in the regenerated plants. FCM, SSR and CAPS demonstrated that intergeneric diploid cybrids have been successfully obtained by symmetric fusion. Related results concerning nuclear and cytoplasmic composition of previous diploid somatic hybrids and potential mechanism for regeneration of such kind of plants are discussed herein. This revised version was published online in July 2006 with corrections to the Cover Date.     

Clonal propagation of F1 hybrids as a tool in genetic studies of the soya bean [Glycine max (L.) Merrill]

Summary A simple technique for the vegetative propagation of F1 hybrid soya bean plants is described. By growing 1- and 2-wk old cuttings, all treated with 1-naphtylacetic acid and the fungicide metalaxyl, average success rates of 18 and 34% respectively were obtained. Cuttings grown under a long-day regime for two months produced large numbers of F2 seeds to be used in genetic studies. This method reduced the time and effort necessary to obtain hybrid progeny.     

Height,competition and yield potential in winter wheat

Summary A monogenic dominant male sterility is used for hybrid production in autumn and winter cauliflower. The ratio of male sterile plants in the backcross progenies of autumn cauliflower was 1:1 over five years (1987–1991). However, a significant deficit of male sterile plants was observed in the winter type over the same period.The influence of the temperature on the male sterile phenotype was studied within backcross progenies planted inside polythene tunnels. Six classes of phenotype were defined during the flowering period (from May to November). At low temperature, some male sterile plants developed partial to complete male fertility, whereas at high temperature, male fertile plants became male sterile.Segregation among the progenies of self-pollinated unstable male sterile plants did not deviate from the expected 3:1 ratio. Plants homozygous for the male sterility allele have been revealed by test crosses with a male fertile plant.For use in seed production, stable male sterile plants are vegetatively maintained; however, crossing lines isogenic except at the MS locus would allow male sterile plants to be raised from seed.     

An instant bioassay for resistance of lettuce to the leaf aphid Myzus persicae

Summary A reliable impression of the resistance of lettuce plants to the leaf aphid Myzus persicae can be obtained via aphid honeydew production. Under controlled temperatures, the number of honeydew droplets produced by these aphids per plant in 180 minutes with five plants per genotype offers a good criterion of this resistance.