Co-ordinated gene expression during phases of dormancy release in raspberry (Rubus idaeus L.) buds

Bud break in raspberry (Rubus idaeus L.) is often poor and uneven, with many of the subapical buds remaining in a dormant state. In order to determine the dormancy status of raspberry buds, an empirical measure of bud burst in a growth-permissive environment following exposure to chilling (4 degrees C cold storage) was developed. For cv. Glen Ample, percentage bud burst in intact canes and isolated nodes was recorded after 14 d. Isolated nodes (a measure of endodormancy) achieved 100% bud burst after approximately 1500 h chilling whereas buds on intact plants (combined endo- and paradormancy) required an additional 1000 h chilling. A microarray approach was used to follow changes in gene expression that occurred during dormancy transition. The probes for the microarrays were obtained from endodormant and paradormant raspberry bud cDNA libraries. The expression profiles of 5300 clones from these libraries were subjected to principal component analysis to determine the most significant expression patterns. Sequence analysis of these clones, in many cases, enabled their functional categorization and the development of hypotheses concerning the mechanisms of bud dormancy release. Thus a set of novel candidates for key dormancy-related genes from raspberry buds have been identified. Bud dormancy is fundamental to the study of plant developmental processes and, in addition, its regulation is of significant economic importance to fruit and horticultural industries.     

In nature dormant buds and in vitro dormant-like buds ofFraxinus excelsior L.

Summary Microcuttings ofFraxinus excelsior, sampled from adult trees during the period of cell cycle blockage of bud in G_0–1, developed long rejuvenated sprouts on the Woody Plant Medium (WPM) with benzylaminopurine (BAP) (4.0 mg/l) and indolebutyric acid (IBA) (0.03 mg/l). These sprouts had the ability to enter a resting period, building dormant-like buds when maintained on the original WPM. Sprouts developed from subcultures also entered a resting period without any transfer. Comparison of in nature buds in active growth and dormancy with buds of growing sprouts and in vitro dormant-like buds revealed similarity in behaviour at the shoot apical level. In particular, in dormant-like buds in a constant environment, shoot apical functioning was suppressed while the cell cycle of the shoot apex was blocked at the G_0–1 phase, like in nature dormant buds.Abbreviations a.u. arbitrary unit - BAP benzylaminopurine - CF cumulative frequency - d₁, d₂ diameters of the shoot apex - IBA indolebutyric acid - P_n last opposite primordia of range n - WPM woody plant medium     

Chromosome numbers and reproduction inRubus subgen.Malachobatus

A review of current knowledge of chromosome numbers and modes of reproduction in the genusRubus L. is presented. Chromosome numbers from some species of subg.Malachobatus Focke together with results of crossing experiments are reported for the first time.     

Accessory Buds in the Genus Rubus with Particular Reference to R. idaeus L.

Accessory (secondary) axillary buds occurred in 21 out of 32Rubus species, including wild raspberry, R. idaeus L. The adaptivevalue of the secondary-bud character is discussed. Disbuddingexperiments on three raspberry cultivars and their inbred derivativesrevealed considerable variability in proportion of secondarylaterals developing and in numbers of flowers per secondarylateral, suggesting the possibility of selection for high secondarycrop potential.     

The inheritance of chemical phenotype in Cannabis sativa L

Four crosses were made between inbred Cannabis sativa plants with pure cannabidiol (CBD) and pure Delta-9-tetrahydrocannabinol (THC) chemotypes. All the plants belonging to the F(1)'s were analyzed by gas chromatography for cannabinoid composition and constantly found to have a mixed CBD-THC chemotype. Ten individual F(1) plants were self-fertilized, and 10 inbred F(2) offspring were collected and analyzed. In all cases, a segregation of the three chemotypes (pure CBD, mixed CBD-THC, and pure THC) fitting a 1:2:1 proportion was observed. The CBD/THC ratio was found to be significantly progeny specific and transmitted from each F(1) to the F(2)'s derived from it. A model involving one locus, B, with two alleles, B(D) and B(T), is proposed, with the two alleles being codominant. The mixed chemotypes are interpreted as due to the genotype B(D)/B(T) at the B locus, while the pure-chemotype plants are due to homozygosity at the B locus (either B(D)/B(D) or B(T)/B(T)). It is suggested that such codominance is due to the codification by the two alleles for different isoforms of the same synthase, having different specificity for the conversion of the common precursor cannabigerol into CBD or THC, respectively. The F(2) segregating groups were used in a bulk segregant analysis of the pooled DNAs for screening RAPD primers; three chemotype-associated markers are described, one of which has been transformed in a sequence-characterized amplified region (SCAR) marker and shows tight linkage to the chemotype and codominance.     

The inheritance of genetic markers in microspore-derived plants of barley Hordeum vulgare L.

Summary Biochemical, molecular and morphological markers have been used to monitor the segregation of alleles at major gene loci in microspore-derived lines of four spring barley crosses and their parents. Significant deviations from the expected Mendelian ratios were observed for four of the ten markers studied in the cross. Distorted ratios were associated with loci located on chromosomes 4H and 6H. The differential transmission of alleles was in favour of the responsive parent (Blenheim) used in the anther culture studies. For the -Amy-1 locus on chromosome 6H, the preferential transmission of Blenheim alleles was most pronounced in the haploid regenerants that were colchicine treated. These results are discussed in relation to the genetic control of androgenetic response in barley and with respect to the exploitation of another culture in barley improvement.     

The Histology of Seed Development in Diploid and Tetraploid Raspberries (Rubus idaeus L.)

Ovules of the raspberry, Rubus idaeus L., resembled those ofother Rubus species in being unitegmic, crassinucellate, andanatropous. In one cultivar, ‘M. 69/139’, both eggcell and embryo were inserted slightly laterally to the micropyle.Of the tetraploid forms, 4n M. 69/139 showed a proportion ofembryo sacs with delayed differentiation, and 4n ‘M. Jewel’some with four antipodal cells. Embryo development showed littleuniformity in timing, but followed the Asterad type. Occasional delayed endosperm development seemed to be a varietalpeculiarity of M. 69/139. At maturity, the ergastic materialin healthy endosperms consisted of oil globules and aleurongrains. The hypostase formed a projecting pedestal below theendosperm at 8 days. The inner epidermal layer of the testaappeared specialized, both in the early stages of seed growth,when it is suggested that it functioned as an endodermis, andat maturity, when it appeared to form an osmotic barrier.     

Polymorphism and inheritance of gliadin polypeptides in T. monococcum L.

Summary More than 80 different gliadin electrophoretic patterns (spectra) have been found in 109 accessions of the diploid wheat Triticum monococcum. Each pattern consists of 15–20 gliadin bands. Some patterns are clearly related and might arise from one another through single mutations in the gliadin-coding loci. From the analysis of 15 grains of each, only 61 accessions were found to be uniform; others consisted of two or more grain variants differing in their gliadin spectrum. An analysis of F₂ grains from three crosses between different accessions showed that groups (blocks) of components are jointly and codominantly inherited. Two independent major Gli loci were established. The close resemblance of the composition of some blocks of T. monococcum to some of those in polyploid wheats indicates that one locus in each T. monococcum genotype is located on chromosome 1A (Gli-A1) and the other on 6A (Gli-A2). However, the blocks of T. monococcum include more bands than corresponding (equivalent) blocks of polyploid wheats. Two out of 275 F₂ grains of the cross k-14244 x k-20409 were found to have gliadin spectra which can be explained as a result of intralocus recombination. Also, a second gliadin-coding locus on chromosome 1A was found in the cross k-46140 x k-46753. This locus recombines with the main Gli-A1 locus with a frequency of about 22% and was clearly analogous to the additional Gli locus found earlier on chromosome 1A of certain polyploid wheats.     

Induction of adventitious buds in cultured petal explants ofChelidonium majus L.

Petal explants ofChelidonium majus L. (Papaveraceae) formed noteworthy adventitious buds without any intermediate callus when cultured under appropriate conditions. Bud formation was favored by combinations of 1–2 mg/l indoleacetic acid (IAA) and/or 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.1–0.5 mg/l kinetin (K). In the present study, neither bud formation nor callus formation occurred in cultures of excised leaves. A histological study revealed that adventitious bud formation occurred only in single epidermal layers of petals, while several subepidermal parenchyma layers did not join in its formation. Activation zones arising from the epidermis underwent intense cell divisions to initiate buds on the epidermal surface. These buds later turned green in color, developing into shoots which eventually grew into plantlets after root formation.     

Interaction of endodormancy and paradormancy in raspberry (Rubus idaeus L.)

Bud break in protected Northern European raspberry crops is often poor and uneven with many of the sub-apical buds remaining in a dormant state. In order to improve bud break and therefore yields, the mechanism controlling bud dormancy must be determined. Canes of the biennial cultivar ‘Glen Moy’ were forced as isolated single nodes, trisections, or as intact canes after different lengths of cold storage chill unit (CU) accumulation in order to determine whether the buds were in an endodormant or paradormant state. The results showed that buds on the lower parts of the intact canes remained in a dormant state long after buds from higher up the intact cane and also the single nodes from all parts of the cane had emerged from the deepest phase of endodormancy. This would imply that these buds were being held in a paradormant state until large amounts of chilling units (>1000 CU) had been accumulated. The trisected cane portions revealed almost no significant differences in bud break levels throughout the experiment when compared with the single nodes. This suggests that removal of the apical part of the cane would be effective in improving bud break by reducing the paradormant condition. A period of secondary dormancy was also observed in the intact canes which may also exacerbate the poor bud break observed in protected crops. This was not seen in the single nodes or the trisected canes which indicates that treatments which reduce paradormancy may also minimise the risk of secondary dormancy. By identifying the phase of bud dormancy which causes poor bud break, attention can now be focused on methods which overcome paradormancy in protected crops. Such methods might include tipping (removal of the cane apex), horizontal training methods, more efficient chilling methods, and chemical treatments.     

The Histology of Seed Development following Crosses between Diploid and Autotetraploid Raspberries (Rubus idaeus L.)

Seed development was studied in crosses made between the diploidforms and autotetraploid mutants of two raspberry (Rubus idaeusL.) cultivars. Drupelet abortion did not appear to be causedby failure of fertilization; it occurred mainly between thefourth and eighth day after pollination, and manifested itselfas simultaneous cessation of growth in all tissues. In healthydrupelets at eight days after pollination, growth appeared tobe proceeding at similar rates in all the tissues except thatthe embryo seemed to be growing faster than the endosperm inthe most advanced crosses. Both diploid and tetraploid ovulesappeared to be more stimulated by pollen from tetraploid plants.Embryo differentiation differed according to the direction ofcross, especially in stunted embryos. Radicles tended to beabnormally large when diploid pollen was used and smaller thanusual with haploid pollen.     

Plastid inheritance in Job's tears,Coix lacryma-jobi L.

Summary The yellow striping in Job's tears showed nonchromosomal maternal inheritance of the trait producing green, yellow lethal and striped seedlings in the offspring in widely different ratios whenever the striped plant was used as female parent.Plastids in yellow regions of the striped plant were of various sizes and colours ranging from normal to minute size and light yellow or pale green to transparent ones. They were even absent in some cells. The plastids in the leaves of the yellow lethals were usually pale green.While the histological studies indicate the probability that the genetic determinants affecting plastids were in the cytoplasm, the possibility that all the abnormal plastid types could be different expressions of the same original mutation that occurred in the plastids, together with the extensive data on the breeding behaviour of the striped plant, is strongly in favour of the concepts of plastid mutation and autonomy.     

The role of female accessory glands in maternal inheritance of phase in the desert locust Schistocerca gregaria

Accessory glands of crowd-reared females of Schistocerca gregaria were ligatured from the lateral oviducts. Hatchlings resulting from egg pods laid after the treatment showed a significant shift towards solitarious behaviour as compared to hatchlings from control-treated females. Morphometric measurement of hatchlings revealed no consistent difference between ligatured and control females, however, one ratio (hind femur length/vertex width) was approaching significance. Hatchlings from eggs of crowd-reared females behaved solitariously when freshly laid eggs were washed with a saline solution. Gregarious behaviour could be restored when washed eggs were treated with a saline extract of the accessory glands. The colouration of hatchlings was not affected by any treatment. Our findings implicate the accessory glands in the production, release or activation of the recently reported gregarizing factor found in the egg pod foam [McCaffery, A.R., Simpson, S.J., Islam, M.S., Roessingh, P., 1998. A gregarizing factor present in the egg pod foam of the desert locust Schistocerca gregaria. Journal of Experimental Biology 201, 347-363].     

Propagation of Morus indica L. (Mulberry) by encapsulated shoot buds

Axillary buds of mulberry (Morus indica L) were encapsulated in alginate and agar to produce individual beads. The beads could be stored at 4°C for 45 days without loss of viability. Amongst the encapsulating agents tested, sodium alginate was found to be a better matrix. Encapsulated buds regenerated complete plantlets on an appropriate medium. This technique would provide an easy and novel propagation system for the elite as well as difficult-to-root species of mulberry.Abbreviations BAP benzylaminopurine - MS Murashige and Skoog (1962).     

Changes of morphogenic competence in mature Pinus sylvestris L. buds in vitro

The effects of season and cold storage on morphogenic competence in mature Pinus sylvestris buds were investigated. Peroxidase and polyphenol oxidase activity were measured as markers of oxidative metabolism. No growth in vitro was observed on explants detached from the end of January until the beginning of March. Brachioblasts, each with a couple of needles, formed on 11% of the buds without macrostrobili that were detached in early April and introduced immediately into culture. Of the explants detached in late July, 15% formed shoots with brachioblasts and needles. The lowest activity of peroxidase and polyphenol oxidase in pine buds was observed from the end of April until the beginning of June when morphogenic competence of tissues started to increase. Development of bud explants detached in January was achieved by cold storage for 5 months. Low polyphenol oxidase and peroxidase activity coincided with increased morphogenic potential. Results suggest that reduced or stable activity of peroxidase and polyphenol oxidase is associated with an increased ability of tissues to start growth in vitro.