The influence of cytokinins and ionic strength of Anderson's medium on shoot establishment and proliferation of evergreen azalea

Shoot-tip explants of evergreen azalea cv. Fuchsia grown on Anderson's medium and containing different cytokinins produced the highest proliferation rate on a medium containing thidiazuron (TDZ). TDZ concentrations ranging from 0.23 to 2.3 M resulted in both good bud-break rate (4 to 5) and shoot quality (> 0.5 cm in length). Adding 2.3 M zeatin to Anderson's medium containing 0.23 or 2.3 M TDZ increased the number of axillary shoots/explant. However, increasing the zeatin concentration to 4.6 M resulted in a reduced shoot proliferation rate. A medium containing 1.15 M TDZ and 2.3 M zeatin resulted in an 18-fold increase for 'Fuchsia' and a 9-fold increase for 'Hino Crimson' after 6 weeks of culture. It was found that explants grown on a half-strength Anderson's medium with 87.6 mM sucrose generally had better shoot proliferation rate and shoot quality than at higher ionic strength.     

Aiuminium tolerance of durum wheat germplasm

Summary Aluminium tolerance of Macedonian durum wheat (Triticum turgidum L. conv. durum (Desf.) MacKey) germplasm was evaluated in nutrient solutions containing 0, 74 or 148 M of total Al. Relative root length (148 M Al/0 Al) of various genotypes ranged from 41 to 72% (from moderately sensitive to moderately tolerant to Al). No genotype with Al tolerance close to that of very tolerant T. aestivum cultivar Atlas-66 was found. Seed Ca concentration was positively (r=0.64, P0.05) and seed Fe concentration negatively (r=–0.71, P0.05) related to the relative root growth. Such a significant correlation was not obtained for seed concentrations of other nutrients or seed protein content.Abbreviations HSD Tukey's Honestly Significant Difference - RRL-2 relative root length, in % (74 M total Al/0 M Al) - RRL-4 relative root length, in % (148 M total Al0 M Al)     

Effects of salicylic acid on cold preservation and cryopreservation of encapsulated embryonic axes of Persian lilac (Melia azedarach L.)

Embryonic axes of Persian lilac (Melia azedarach L.)encapsulated into calcium alginate beads with sucrose (0.75 M) and salicylicacid (0, 50 M, 200 M) were subjected to cryopreservationtechnique with dehydration and freezing in liquid nitrogen or to coldpreservation by stocking alginate beads in empty petri dishes for 4 monthsat 4 °C. In these two cases 200 M salicylic acid enhancedsignificantly the percentage of viability of encapsulated embryonic axes andthe role of salicylic acid in increasing tolerance to dehydration is discussed.     

Pollen grain size in four ploidy levels of genusAvena

Summary Pollen grain size in many genera positively correlated with chromosome number. In this study, oat (Avena) pollen grain size was examined in diploids (2×=14, one species, for accessions), tetraploids (4×=28, five species, 20 accessions), hexaploids (6×=42, one species, eight cultivars), and in 10 octoploid (8×=56) accessions. Mature anthers ready to dehisce pollen were sampled from one to six plants per accession, and pollen grains were squeezed out of the anther with tweezers. Oat pollen grains are slightly elliptical, and the length of the major axis was found to be highly correlated with the minor axis width (r=0.94^**). Pollen grain length, 39.3 m for diploids, 41.3 m for tetraploids, 47.0 m for hexaploids, and 48.8 m for octoploids, was positively correlated (r=0.86^**) with ploidy level. No genomic or species effects appeared to influence this trait. Only tetraploidAvena vaviloviana accession PI 412767 produced two distinct class sizes of pollen grains, 99% normal (43.0 m) and 1% large (52.7 m).     

High frequency somatic embryogenesis with a pampeana-derived genotype of alfalfa (Medicago sativa L.)

Summary Somatic embryos of genotype R11 of the alfalfa variety Pampeana were produced from embryogenic calli derived from leaf sections. They were induced by an auxin shock and its development was attempted on six different media. The best condition for somatic embryo production was inducing callus on MS medium plus 10 M 2,4-D and 4,6 M KIN and transferring them, after the auxin shock, to MS with 10–20 mM NH₄ ⁺ and 30 mM proline. More than 500 somatic embryos per plate were produced. Embryos were grown to plants on MS or half strength MS media and all regenerated plants resembled the original R11 genotype. This technique could be useful in alfalfa Pampeana improvement using genetic modification.     

Interspecific hybrids of Cucumis metuliferus × C. anguria obtained through embryo culture and somatic embryogenesis

Summary Interspecific crosses between Cucumis metuliferus Naud. and C. anguria L. were obtained through embryo culture. Embryos in the rabbit-ear to advanced fluke-shaped stages were rescued 34–99 days after pollination. Plants were obtained through direct embryo culture, and through somatic embryogenesis from immature embryos. For direct embryo culture, fluke-shaped embryos were stored in sterile water in darkness for three days at 25C prior to transfer on Murashige and Skoog (MS) culture medium plus 1.0 M 6-benzylamino-purine. Multiple plants were obtained from single embryos through somatic embryogenesis of rabbit-ear stages on MS plus 10 M indole-3-acetic acid and 5 M 6-benzylamino-purine. Evidence of hybridization included leaf shape intermediate between the two parents, penduncle shape prior to fertilization which resembled the male parent, low pollen viability and isoelectric focussing of protein bands for acid phosphatase of leaf extracts.     

Studies on glucosinolates in clubroot resistant selections and susceptible commercial cultivars of cabbages

Summary Heads of 59 commercial cabbage cultivars, all susceptible to clubroot disease, and of 86 individual clubroot resistant cabbage plants of various breeding selections were analysed for their composition in glucosinolates, determined by quantifying their hydrolytic breakdown products, thiocyanate, goitrin, and volatile isothiocyanates. The mean thiocynate ion content was significantly lower in the breeding selections (120 g/g dry weight) than in the commercial cultivars (204 g/g). In contrast, the mean goitrin content was significantly higher in the breeding selections (193 g/g) than in the commercial cultivars (35 g/g). Similar to goitrin, the range of volatile isothiocyanates and total glucosinolate were higher in the breeding selections, but the mean contents of each were not statistically different between selections and cultivars. Fourteen cultivars and four selections were found to be free of goitrin; three cultivars, but no breeding selection were free of volatile isothiocyanates. The breeding selections will provide germplasm for breeding new clubroot resistant and low glucosinolate cultivars.Contribution No. J-965.     

Somatic embryogenesis and plant regeneration of Tripsacum dactyloides L.

Summary This article reports the culture and plant regeneration of Tripsacum dactyloides. Mature embryos of Tripsacum dactyloides dactyloides were used to obtain embryogenic callus cultures. Currently, 180 normal plants have been regenerated from these cultures. Callus was initiated on MS medium supplemented with dicamba (10 mol or 20 mol) and sucrose (3% or 6%), and plants were regenerated on hormone free MS medium containing 2% sucrose. No significant differences were found in callus initiation frequency or in embryogenic response of cultures on the four combinations of sucrose and dicamba tested. The embryogenic cultures have been maintained for 9 months (12 subcultures) and have retained regeneration capacity. Plants regenerated from tissue culture of maize-by-Tripsacum hybrids could be useful in maize improvement.     

Colchicine, trifluralin, and oryzalin promoted development of somatic embryos in Ilex paraguariensis (Aquifoliaceae)

A protocol for somatic embryogenesis and plant regeneration of Ilexparaguariensis St. Hil. from embryos cultures was developed. Heart stage zygotic embryos were removed from seeds of immature, light green fruit and treated with antimicrotubule agents (0.1; 0.2, and 0.5% colchicine for 24 and 48 h; 1; 10, and 20 M of either trifluralin, - trifluoro- 2,6-dinitro-N,N- dipropyl-p-toluidine, or oryzalin, 3,5-dinitro-N₄, N-dipropylsulphate during 48 h). The embryos were cultured aseptically on quarter-strength Murashige and Skoog medium containing 3% sucrose, 0.65% agar (1/4MS), and 0.46 M zeatin. Cultures were incubated in darkness at 27 ± 2 °C. All thetreatments provoked a diminution of the number of germinated embryos and in some of the treated embryos somatic embryogenesis was induced. Somatic embryo maturation and conversion into whole plants could be achieved by culturing the embryos on 1/4MS lacking hormones and incubated at 27 ± 2 °C, 14 h photoperiod (116 mol m^-2s^-1). Mostof the plants regenerated from somatic embryos appeared morphologically normaland grew under greenhouse conditions. Only 2 plants out of 152 studied contained the tetraploid number of the chromosomes (2n = 4x = 80), meanwhile the rest of the plants had the normal diploid number of chromosomes (2n =2x = 40). Somatic embryos with abnormal morphology were also observed.     

A tissue culture technique for vegetative propagation and low temperature preservation of Beta vulgaris

Summary A tissue culture technique for clonal propagation of Beta vulgaris is described. Flower buds or flower bud clusters formed adventitious shoots on half-strength Murashige & Skoog's medium supplemented with 10 mol/l benzyladenine (BA). The shoots were multiplied by axillary bud proliferation on a medium with 1 mol/l BA and rooted on a medium with 10 mol/l indolebutyric acid. The plants were vegetative. No mutations were observed. Genotypic variation was found in shoot formation, shoot multiplication and rooting. Some genotypes showed leaf malformations which were attributed to BA. Rooted plants in culture tubes could be stored for one year at 2 or 5°C and a low light intensity.     

Genotypic differences in wheat Al tolerance

Summary Aluminium tolerance of 83 genotypes from Croatian and Yugoslav Triticum aestivum germplasm was evaluated in nutrient solutions having Al³⁺ activities of 0, 12.5 and 25 M. Relative root length (25 M Al³⁺/0 Al) of various genotypes ranged from 2 to 97% (from very sensitive to tolerant to Al). No genotype with Al tolerance close to that of very tolerant cultivar Atlas-66 was found. Soil, climatic, fertilization, and liming effects that wheat plants giving seeds for the nutrient solution Al-tolerance screening had been subjected to during their growth cycle did not influence the Al-tolerance ranking. Significant correlation was found between screening wheat for Al tolerance in nutrient solutions and in acid Pseudogley soil amended with five rates of limestone in a greenhouse experiment. Seed protein concentration was significantly related to the Al-tolerance ranking (r² = 0.962). Such a significant correlation was not obtained in a case of rheological and other quality characteristics of seeds. Al-tolerant wheat genotypes identified in this study will be used in breeding for improved Al tolerance.Abbreviations HSD Tukey's Honestly Significant Difference - RRL-2 relative root length, in % (12.5 M Al³⁺/0 Al) - RRL-4 relative root length, in % (25 M Al³/0 Al)     

In vitro adventitious shoot regeneration for effective maintenance of male sterile niger Guizotia abyssinica (L.f.) Cass.)

A rapid and efficient method of in vitro plant regeneration for large scale propagation of male sterile plants of niger (Guizotia abyssinica) was developed. Leaf segments from mature plants were cultured on Murashige & Skoog's basal medium (MS) supplemented with N⁶-benzyladenine (BA) and kinetin individually and in combination with low concentrations of indole-3-butyric acid (IBA); and -Naphthaleneacetic acid (NAA). Prolific direct adventitious shoot regeneration occurred on most of the media tested. The best response in terms of frequency of shoot regeneration and the number of shoots per leaf explant was observed on medium supplemented with 2.22 M BA. Transfer of shoot bud clusters to fresh medium with same composition promoted further multiplication of the shoot buds, while medium with reduced BA concentration (0.89 M) facilitated shoot elongation. Shoots that were rooted on half-strength MS medium gelled with 0.2 or 0.4% agar and supplemented with 4.9 M IBA survived with a frequency of 61.36% on transfer to ex vitrum conditions. Field evaluation of the regenerants revealed the genetic stability of the plantlets and are being used in breeding of experimental hybrids.     

Evaluation of Triticum aestivum germplasm from Croatia and Yugoslavia for aluminium tolerance

Summary Aluminium tolerance of 90 genotypes of Triticum aestivum L. germplasm from the breeding programmes of eight Croatian and Yugoslav institutions was evaluated in nutrient solutions having Al³⁺ activities of 0, 12.5 and 25M. Overall distribution of Al tolerance of wheat genotypes was skewed toward lower tolerance rankings. Average Al tolerance differed among gene pools created at different breeding institutions. Genotypes tolerant enough to be useful in the breeding programmes aimed at selecting cultivars with improved Al tolerance are identified in germplasm from four institutions. No correlation was found between chemical characteristics of soils used over the years by breeding institutions for their field trials and the Al-tolerance ranking of the corresponding germplasm material.Abbreviations HSD Tukey's Honestly Significant Difference - RRL-4 relative root length, in % (25M Al³⁺/0 Al)     

Plant recovery of cryopreserved apical meristem-tips of Melia azedarach L. using encapsulation/dehydration and assessment of their genetic stability

A cryopreservation process usingencapsulation/dehydration technique was setup for apical meristem-tips of invitro plantlets of `paradise tree' (Melia azedarach L. var. gigantea,clone `El dorado'). Apical meristem-tipswere cultured for one day on MS basalmedium with 2 M BA and 0.5 M IBAand encapsulated with 3% sodium alginate.The highest shoot proliferation rate aftercryopreservation was obtained whenencapsulated apical meristem-tips werepregrown for 3 days in liquid medium with0.5, 0.75 and 1 M of sucrose for 24 hoursprogressively, desiccated for 5 hours withsilicagel followed by rapid or slowcooling. Survival after freezing in liquidnitrogen ranged between 67–83% andshoot proliferation ranged between 43–60%. This cryopreservation treatmentpreserved genetic stability, when it wasevaluated using the electrophoreticpatterns of nine isozyme systems and RAPDprofiles.     

Interspecific hybridization between Abelia × grandiflora ‘Francis Mason’ and A. schumannii via ovule culture

Ethiopian mustard (Brassica carinata Braun) is a potential oil crop for the Mediterranean area. The objective of this study was to develop an efficient system of mutagenesis using ultraviolet (UV) light irradiation of isolated microspores from Brassica carinata. From the survival curve based on embryo yield after irradiation of the microspores with UV light, the LD50 was estimated to be an exposure of 8 min. Total content of glucosinolates and fatty acid composition were analysed in the seeds of the doubled haploid homozygous plants with the purpose of selecting lines with modified glucosinolate and erucic acid contents. Three groups of doubled haploid lines exhibiting low and high glucosinolate contents, and high erucic acid content have been identified from a population of 270 doubled haploid lines. In eight lines, the content of glucosinolates was reduced from an average of 80.6 mol g^-1 seed to 37.5 mol g^-1 seed, whereas in four lines, the content of glucosinolates was increased up to 99.2 mol g^-1 seed. In six additional lines, the content of erucic acid was increased from 42.8% in the nontreated lines to 49.5% of the totalfatty acid composition in some of the mutant lines. All lines showed stablelevels of erucic acid in two generations, the M₂ and M₃.     

Bolting resistant breeding of Chinese cabbage. 1. Flower induction of late bolting variety without chilling treatment

Summary When a local slow bolting variety Osaka Shirona Bansei (Brassica rapa L. ssp. pekinensis, syn. B. campestris L. ssp. pekinensis) was grown in a phytotron (25°C, 16 hours day length without chilling treatment), one third of the plants bolted and flowered. In order to clarify the different flowering responses in the variety, a progeny line (FNC) of the flowering plants was chilled for 4 different periods (0, 22, 36 and 53 days) in a chamber of 2 7°C, then transplanted to three different conditions, i.e. PHY: 25–20°C day and night temperatures, 16 hours day length, GHL: 10 25°C, 16 hours day length with supplementary light and GHN: 10 25°C, natural day length (10 15 hours). In PHY, FNC bolted and flowered with almost the same leaf numbers in all 4 different chilling treatments. This means that FNC has very low sensitivity and no requirement to low temperature for its reproductive growth. In GHN (short day length), FNC bolted very slowly. Then the bolting and flowering of FNC were promoted by both long day length and high temperature. The newly found bolting characteristics of Osaka Shirona Bansei could be applied to breed unique slow bolting Chinese cabbage (B. rapa L. ssp. pekinensis) which might be non-sensitive to low temperature and its bolting and flowering would be induced with the combination of long day length and high temperature. Using the unique variety, it might be also possible to establish a new cropping type of Chinese cabbage (late autumn sowing, spring harvest).Abbreviations PHY Phytotron, long day and high temperature condition - GHL Greenhouse, long day and medium temperature condition - GHN Greenhouse, natural (short) day and medium temperature condition - FNC A progeny line of Osaka Shirona Bansei which flowered with no chilling treatment - FC A progeny line of Osaka Shirona Bansei which flowered with chilling treatment     

Genetic analysis of hexaploid wheat,Triticum aestivum using intervarietal chromosome substitution lines—protein content and grain weight

Summary The 21 intervarietal chromosome substitution lines of the cultivar Hope in Chinese Spring were used to analyse the genetic differences between the two cultivars Hope and Chinese Spring in grain protein content and grain weight.Only one chromosome of Hope, 5D, significantly influenced grain protein content of Chinese Spring. Its influence was of only minor effect and was to decrease protein content expression of Chinese Spring. It has been postulated that the genetic control of protein content, in this instance, is most likely due to many genes each of small effect.Five chromosomes of Hope influenced the 1000 grain weight value of normal Chinese Spring, all increasing its expression. Chromosomes 1A, 4A and 5B were of major effect and 3A and 6A of comparatively minor effect. A minimal estimate of five genes determines the difference in grain size between these cultivars. The possible evolutionary significance of the contribution of the A genome of bread wheat to grain size determination is discussed. On the basis of certain findings of this study, proposals are made for breeding for increased grain size in hexaploid wheat.     

Male-sterility in nigerian bulrush millets (Pennisetum americanum (L.) K. Schum)

Summary The occurrence of three male-sterile plants is reported. One in a population of ex-Bormu, an improved recommended variety of day-length neutral gero bulrush millet and the two others in the selfed progeny of a population of maiwa bulrush millet, a short-day photoperiod-sensitive type. Tests confirmed the cytoplasmic-genic nature of the male-sterility in the gero population. No tests could be conducted for maiwa.The transfer of male-sterility from the male-sterile Tift 23A bulrush millet, obtained from the United States, into a maiwa population is also reported. After six backcrosses the maiwa genotype appeared to have been reconstituted against the background of a male-sterility inducing cytoplasm. Maiwa male-sterility maintainer plants have also been identified.     

Resistance to the glasshouse whitefly (Trialeurodes vaporariorum Westw.) in sweet pepper (Capsicum annuum L.)

Summary Fourteen cultivars, one F₁-hybrid and one half-wild type (Cind) of sweet pepper were tested on resistance to glasshouse whitefly. Several mutually significant levels of resistance were found. Most resistant appeared to be California Wonder, Severka M, Korál and Yolo Wonder. During the test sweet pepper plants were infested both with whiteflies and their parasite — Encarsia formosa, which caused the blackening of whitefly puparia. The blackened puparia are easily seen, enabling an easier and more rapid evaluation of the number of puparia present.     

Chemically induced sterility in wheat for hybrid seed production

Summary The gametocidal effects of RH-531, RH-532, and RH-2956 were studied with three rates and times of foliar application on two spring wheat (Triticum aestivum L.) varieties, Anza and Yecora 70. RH-531 and RH-532 applied at or before booting stage caused rather large reduction in plant height while RH-2956 had a small effect on height. Application of any of the three chemicals on Anza at meiotic stages reduced self-pollination (SP) fertility by 97–99%. If 90% SP fertility reduction is acceptable, the application time could vary from premeiosis to postmeiosis for Anza. Yecora 70 was less sensitive than Anza to gametocide treatments. The lowest SP fertility obtained in Yecora 70 with RH-531, RH-532, and RH-2956 was by treatment at booting (4 kg/ha), at premeiosis (2 kg/ha), and heading (8 kg/ha), and the corresponding SP fertility reductions were 74, 89, and 71%. Pollen stainability-determined by aceto-carmine, IKI, and Alexander's stains-was not affected by gametocide treatment and was not useful in evaluating sterility induced by these chemicals. For the RH-531-treated Anza and Yecora 70 and the RH-532-treated Anza, more than 94% of the seeds produced by open-pollination on treated plants were from self-or sib-pollination as determined by progeny testing. However, RH-532-treated Yecora 70 and RH-2956-treated Anza and Yecora 70 did result in more crossed seeds, with a maximum of 20% outcrossing. It was believed that spike compactness induced by the chemicals and the related poor flower opening were the major limitations for outcrossing. Artificial pollinations made to the test varieties showed that RH-531 induced both male and female sterility and was undesirable for practical use. RH-532, however, could be applied at the booting stage to avoid severe female sterility, and RH-2956 clearly was the best of the three chemicals, with the female fertility very close to those of the untreated checks of both Anza and Yecora 70. Of the three chemicals, RH-2956 gave the highest percentage of hybrid plants in the progeny of hand-or open-pollinated spikes. Its practical use for hybrid seed production, however, will depend largely on improvement of methods to increase cross pollination in wheat.