Clonal propagation of Triticum durum Desf. from immature embryos and shoot base explants

Summary Immature embryos from five durum wheat cultivars were grown on Murashige and Skoog medium supplemented with two concentrations of kinetin or 6-benzylaminopurine (BAP). The embryos cultured on the medium containing 5 mg/l of BAP proliferated several axillary shoots. Shoot base segments subcultured on the same medium gave more shoot proliferation. The shoots developed into ear-bearing plants. This technique could be used for clonal propagation of wheat.     

Clonal propagation of rice through proliferation of axillary shoots

Summary Shoot base segments have been explanted from seedlings of rice (Oryza sativa L. subsp. Japonica, cv. Arborio) and grown on agar-solidified MS medium supplemented with different concentrations of four cytokinins: kinetin, BAP, 2iP and zeatin. After one month, segments were explanted from proliferated shoots and subcultured on their respective media. BAP was by far the most effective in inducing shoot proliferation. Highest rates were achieved at the higher concentration used: 5 mg 1^–1. Shoot base segments were subcultured fifteen times consecutively on seven different concentrations of BAP. Shoots grown in the presence of 5 mg 1^–1 of BAP proliferated an average of 12 normal shoots for each base segment throughout the fifteen subcultures. The shoots rooted easily on hormone-free medium. The technique does not require any particular skill, it is very effective and, therefore, can be suggested as suitable for clonal propagation of rice.     

The response of a range of genotypes of Vitis vinifera to sequential shoot tip cultures at high temperatures

Summary The evaluation of genetic variation and the release of virus-free cultivars are important in Vitis breeding. Sequential shoot tip cultures and high temperatures (32°, 35°, 38°C) were used to study their effects on the rate of shoot elongation during the proliferation phase (initial stage) of the in vitro culture in some Vitis vinifera cultivars. In the first culture the shoot elongation was more evident at 32°C than at 25° or 35°C; in addition at 35°C it was observed the shoot tip death in some cvs. while at 38°C in all cvs. In the sequential shoot tip cultures we noted a decline on shoot elongation and also an inhibition of shoot morphogenesis in the 4th and 5th recultures.     

Isolation of a pure thornless loganberry by meristem tip culture

Summary Thornless Loganberry (TL) is a periclinal chimeral blackberry in which a layer of mutant (thornless) epidermis surrounds a core of wild-type (thorny) tiusse. Due to its chimeral arrangement, TL produces thorny adventitious root cuttings and thorny offspring. To separate the chimera into its components parts, meristems of TL were grow in vitro on modified Murashige and Skoog medium to yield callus and adventitious shoots. One of these shoots has survived, flowered, and produced thornless offspring from seed. The importance of this non-chimeral TL is discussed.List of terms BA 6-benzylaminopurine - GA₃ gibberellic acid - NAA naphthalene acetic acid - hybridberry polyploid bramble interspecific hybrids - MS Murashige & Skoog (1962) high mineral salt medium - TL Thornless Loganberry - TL_tc tissue culture-derived (non-chimeral) Thornless Loganberry     

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.     

Cryopreservation of shoot tips of Chrysanthemum morifolium and related species native to Japan

Summary The shoot tips of Chrysanthemum morifolium (syn. Dendranthema grandiflorum) and related species native to Japan were cryopreserved using preculture for 2 days, slow cooling (0.2°C/min) until –40°C with 10% dimethyl sulphoxide and 3% glucose prior to immersion into LN₂ and rapid thawing. High survival rates were observed in 3 cultivars of chrysanthemum, 12 species and 2 interspecific hybrids, and slightly low survival rates in 3 species. The shoot regeneration rates of the frozen shoot tips varied from 9.4 to 100% depending on species. Shoot tips of chrysanthemum showed high viability even after a storage of 8 months in LN₂. The thawed chrysanthemum shoot tips grew and flowered normally in a greenhouse under natural conditions.Abbreviations BA 6-benzylaminopurine - DMSO dimethyl sulphoxide - NAA 1-naphtalenacetic acid     

南果梨茎尖离体快繁技术研究

为了探索一种南果梨离体培养快速繁殖的方法,以南国梨茎尖为材料,以MS基本培养基添加6-BA、NAA、IBA进行离体培养及再生体系研究。结果表明最适取材时间为4月上旬;最佳预处理为GA3 0.1 mg/L预处理一周;最佳灭菌时间为0.1% HgCl2溶液灭菌8~10 min;不同品种的南果梨茎尖增殖和生长能力不同;‘大红南果梨’的最适继代培养基为：MS+BA 4.0 mg/L+NAA 0.4 mg/L;IBA促进生根,最适生根培养基为1/2MS+IBA 1.4 mg/L,GA3抑制生根;试管苗移栽的最适基质为珍珠岩:草炭土:河沙=1:1:1,成活率可达93.3%。     

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.     

Effect of in vitro propagation methods on field performance of two strawberry cultivars

Summary Strawberry cultivars, Redcoat and Veestar, propagated by meristem culture (MC), callus culture (CC) and direct shoot regeneration (DR) from leaf disks were compared for their vegetative and reproductive characters with standard runner (SR) propagated plants under field conditions. In the planting year, in vitro propagated plants of both cultivars had the same number of leaves as SR plants, but in vitro propagated Redcoat produced fewer stolons per plant than SR plants. However, in the following year, in vitro propagated mother plants of both cultivars had more leaves and higher runner production than SR mother plants. Flowering and fruiting behaviour of Veestar was not appreciably influenced by in vitro propagation methods. However, in vitro propagated plants of Redcoat flowered earlier and produced more flowers and fruits than SR plants, but still maintained normal berry weight. Among in vitro propagated plants, DR plants of Redcoat were the earliest to flower, whereas MC plants produced more flowers and fruits. The field performance of the first daughter plants derived from the in vitro propagated plants was consistent with their respective mother plants. Leaf shape of both cultivars was not altered by in vitro propagation. Phenotypic abnormalities were mainly confined to occurrence of yellow leaf variants in MC and CC plants and occasional appearance of plants with irregular flowering and growth habit among CC plants.NRCC No. 38004     

Vegetative propagation of Beta vulgaris by leaf cuttings

Summary Leaf cuttings without axillary buds were made from young vegetative beet plants. Roots were formed at the petiole end. Adventitious bud formation occurred at a very low frequency and could not be stimulated by application of growth regulators. Leaf cuttings with axillary buds formed plants at a higher frequency but development of buds into shoots was slow and irregular. As no vascular connections were found between petiole and axillary bud, the slow growth of the buds was attributed to insufficient supply of assimilates from the leaves. Axillary buds rapidly developed into shoots when petiole explants with buds were placed on culture media. Nearly all cultures, however, were contaminated with bacteria that originated from the inner tissue of the explants.     

Mechanisms of resistance to shoot fly, <Emphasis Type="Italic">Atherigona soccata</Emphasis> in sorghum

Summary Sorghum shoot fly, Atherigona soccata (Rondani) is an important pest of sorghum in Asia, Africa, and Mediterranean Europe, and host plant resistance is an important component for the management of this pest. The levels of resistance in the cultivated germplasm are low to moderate, and therefore, it is important to identify genotypes with different mechanisms of resistance to pyramid the resistance genes. We studied the antixenosis for oviposition, antibiosis, and tolerance components of resistance in a diverse array of shoot fly-resistant and -susceptible genotypes. The main plants and tillers of SFCR 151, ICSV 705, SFCR 125, and, IS 18551 experienced lower shoot fly deadhearts at 28 days after seedling emergence, produced more number of productive tillers. The insects fed on these genotypes also exhibited longer larval period (10.1–11.0 days compared to 9.3 days on Swarna), lower larval survival and adult emergence (54.7–67.8 and 46.7–52.2% compared to 73.3 and 60.6% on Swarna, respectively), and lower growth and adult emergence indices as compared to the susceptible check, Swarna. Physico-chemical traits such as leaf glossiness, trichome density, and plumule and leaf sheath pigmentation were found to be associated with resistance, and chlorophyll content, leaf surface wetness, seedling vigor, and waxy bloom with susceptibility to shoot fly and explained 88.5% of the total variation in deadhearts. Step-wise regression indicated that 90.4% of the total variation in deadhearts was due to leaf glossiness and trichome density. The direct and indirect effects, correlation coefficients, multiple and step-wise regression analysis suggested that deadhearts, plants with eggs, leaf glossiness, trichomes on the abaxial surface of the leaf, and leaf sheath pigmentation can be used as marker traits to select for resistance to shoot fly, A. soccata in sorghum.     

Vegetative propagation of Beta vulgaris by leaf cuttings with axillary buds

Summary In leaf cuttings with a short piece of hypocotyl tissue axillary bud development was poor. Most cuttings with a long piece of hypocotyl tissue developed into plants. The latter type enabled vegetative propagation of various genotypes.     

Effect of incubation temperature regimes and culture medium on broccoli microspore culture embryogenesis

Conditions for reliable induction of embryogenesis from isolated microspores were studied in ten genotypes of broccoli. Embryo yields were significantly increased in almost all of the broccoli genotypes by the incubation at 32.5 ^°C for 1 day, than when the standard incubation at 30 ^°C for 2 days was used. Treatments of 48 hours at 32.5 ^°C produced less than optimal results suggesting that broccoli microspores are more sensitive to high temperatures than those of B. napus. The use of the ¹/₂ NLN-13 medium yielded greater number of embryos than the standard NLN-13. The magnitude of the response to the redution of the concentration of major salts by half in the NLN medium varied with the different genotypes. High embryogenic broccoli cultivars, such as ‘Shogun’, ‘SDB9’, and ‘Green Valiant’, presented a better response to the reduction of the concentration of major salts by half in NLN-13. Reduction never produced a detrimental effect on embryo yield and seems not to have any effect in the subsequent development of embryos in plants. This revised version was published online in July 2006 with corrections to the Cover Date.     

The production of homozygous diploid plants of Solanum verrucosum by tissue culture techniques

Summary Monohaploid plants of S. verrucosum (2n=x=12) were induced in anther culture. Axillary buds from these plants were treated with colchicine in shoot tip culture for 48 hours and then transferred to a colchicine free medium. The resulting plantlets were scored for diploidy by stomatal chloroplast counts and root tip cytology and it was found that doubling of the chromosome number had occurred.     

In vitro propagation of Japanese garden iris,Iris ensata Thunb.

Summary Explants of young scapes of Iris ensata were cultured on MS medium with 1 mg/l NAA, 1 mg/l 6-BA, 30 g/l sucrose and 10 g/l agar, and this species was characterized by high variety specificity for callus, shoot and root induction. Among 23 varieties and one wild form tested, Okichidori, Miyukisudare and Meiji-l exhibited a considerable rate of shoot induction, although these induced poorly rooted shoots. In addition, two types of callus induction such as green and white calli were observed, and the induction of green-type calli was significantly correlated with that of shoots. Surprisingly, the only modification, half-strength MS inorganic salts, for the above medium proved to be very effective for shoot induction in the scape culture. For shoots obtained from the scape culture, effects of sucrose concentrations and activated charcoal on root induction were examined by using 1/2 MS with 1 mg/l NAA, 1 mg/l 6-BA, 30 g/l sucrose and 10 g/l agar as the basic medium. The addition of 1% activated charcoal to the media had a marked effect for root induction independent of sucrose concentrations and varieties tested. The in vitro propagation technique of I. ensata is discussed.     

Production of an interspecific hybrid in Gypsophila by ovule-embryo culture

Summary We have succeeded in producing useful interspecific hybrid using ovule-embryo culture between Gypsophila paniculata L. Red Sea and G. manginii, an incompatible combination by ordinary cross breeding methods. The hybrid plant had double flowers with a color of pale purplish pink. Hybrid characteristics of the plant were firmed by observation of plant form, flower type, chromosome number and peroxidase isozyme patterns.     

Somatic in vitro culture response of Lolium perenne L.: genetic effects and correlations with anther culture

Summary Genotypic effects on callus induction and plant regeneration in callus, suspension and protoplast culture, and their correlations with both phenotypic and GCA-values for anther culture response, were studied using 21 genotypes of perennial ryegrass. Differences between genotypes accounted for approximately 40% of the total variation for callus induction and initial callus growth, and 59 and 83% of the variation in callus culture for regeneration percentage and percentage of green plants. Effects of genotypes were less pronounced in suspension culture, where suspensions from the same genotype often behaved differently. Some suspension cultures retained their capacity for green plant regeneration for almost two years, repeatedly producing 80–100% green regenerants during this period. Genotypes with high regeneration percentage and a large proportion of green plants from callus culture were also superior in suspension culture for both regeneration performance and longevity. Regeneration percentage and percentage of green plants were uncorrelated, and probably under different genetic control. While capacity for green plant formation from the different genotypes showed no correlation between anther culture and somatic in vitro culture, a positive correlation was observed between the regeneration percentages in somatic in vitro culture and anther culture (r=0.44^*–0.85^***), suggesting some common genetic control of the two systems.     

High frequency plant-regeneration through direct shoot development and somatic embryogenesis from immature inflorescence cultures of finger millet (Eleusine coracana Gaertn)

Summary Plant regeneration from cultured immature inflorescence segments of Eleusine coracana was obtained by direct shoot development and somatic embryogenesis. Direct development of shoots from cultured inflorescence segments occurred on MS medium supplemented with 2,4-D in combination with zeatin. Inflorescences with well developed spikelets differentiated at a low frequency (<5%) from callus cultures initiated on media supplemented with 2,4-D in combination with zeatin or coconut water or picloram + kinetin. Somatic embryogenesis was also induced in callus cultures growing on MS + picloram + kinetin at the end of four passages. Supplementation of the media with different concentrations of sucrose showed 3% sucrose as the best concentration for plant differentiation from somatic embryos. The majority of the regenerated plants showed the diploid chromosome constitution in their root tips. The regenerants were in general shorter with an increased number of tillers compared to the control.Abbreviations CW Coconut water - 2,4-D 2,4-dichloro phenoxyacetic acid - Kn Kinetin - Z Zeatin     

Interspecific hybrids of Cyclamen persicum Mill. and C. purpurascens Mill. produced by ovule culture

Summary Interspecific crosses were made to introduce the scent of flowers of C. purpurascens into C. persicum cultivars and ovule culture was used to rescue the abortive hybrid embryos. Cultivars of C. persicum diploid (CPD, 2n=2×=48) and C. persicum tetraploid (CPT, 2n=4×=96) were the pistillate parents and wild species of C. purpurascens (CP, 2n=34) were staminate parents. After pollination, crossed ovaries were collected periodically and examined using paraffin sections. Histological observations suggested that both hybrid ovules of CPD x CP and CPT x CP should be transferred to culture medium 35 days after pollination. Based up on this observation, crossed ovaries were collected 28 days after pollination and ovules with placenta were transferred to MS (1962) medium containing 3% sucrose. These ovules were cultured in the dark at 25° C. The hybrids (2n=41) derived from CPD x CP had the scent of C. purpurascens, whereas the hybrids (2n=65) derived from CPT x CP had the scent of C. persicum. Although both hybrids had complete genomes from the parents and produced a few viable pollen grains, they failed to yield viable seeds by self- and cross-pollination with fertile pollen grains of C. persicum cultivars.Abbreviations CPD C. persicum diploid - CPT C. persicum tetraploid - CP C. purpurascens     

Experiments on anther culture in barley. Influence of culture methods on cell proliferation and organ differentiation

Summary Several experiments have been performed in order to induce cell proliferation and plant differentiation from pollen grains by anther culture in barley. Some modifications of the culture media, pretreatments and transfer of the anthers increased notably the frequency of cell division, but the low proportion of normal green plantlets differentiating from microspores and calluses remains the major obstacle preventing the practical use of anther culture in barley breeding.