Chromosome location of genes controlling the content of dietary fibre and arabinoxylans in rye

Differences in fruit-set success reputed to exist between Heveabrasiliensis genetic clones were verified by an analysis of breeding recordsfor the clones PB 5/51, RRIM 600 and PR 107. Although the bestfruit-set success was obtained with PB 5/51 following artificial (hand)pollination, this clonal trait was not reflected in terms of greater numbersof pollen tubes developing it the styles of hand-pollinated PB 5/51 femaleflowers. It was observed, however, that more pollen tubes reached theovules in PB 5/51 than in RRIM 600 or PR 107. Significantly, PB 5/51female flowers required fewer pollen tubes to effect penetration of all threeof its ovules. As fruit formation in Hevea is dependent on all threeovules of the flower being successfully fertilised, PB 5/51 female flowershave hence a greater propensity for successful fruit-set. In all the threeclones studied, the frequency distribution of female flowers with 0, 1, 2 or3 of the ovules penetrated by pollen tubes did not conform to binomialexpectations. Flowers with no ovule penetrated and flowers with all threeovules penetrated were greatly over-represented. One explanation for thisnon-random distribution is the existence of `receptive' female flowers thatfavour successful fertilisation whereas `non-receptive' flowers tend to remainunfertilised even when hand-pollinated.     

The pattern of megasporogenesis and megagametogenesis in diploid Solanum species hybrids; its relevance to the origin of 2n-eggs and the induction of apomixis

Summary Megasporogenesis and megagametogenesis in diploid tuber-bearing Solanum species hybrids were studied and illustrated in detail. Megagametogenesis followed the normal or Polygonum type of embryo sac development. In megasporogenesis a few regularly occurring deviations from what is usually considered to be the normal course of megasporogenesis were detected.Firstly, the development of a surplus of adjacent or non-adjacent megaspore mother cells in a single ovule was frequently observed. As they eventually may give rise to normally reduced, sexual embryo sacs, it is concluded that the archesporium in potato cannot be delimited to a single cell or even a group of adjacent cells in the ovule. Secondly, the micropylar daughter cell which is formed after completion of the first division was often found to degenerate before the onset or completion of the second division giving rise to a triad instead of a tetrad of megaspores. The spatial arrangement of megaspores within the ovule was found to vary according to the variation in the relative orientation of second division spindles.It is concluded that these deviations should be considered random legitimate variations of megasporogenesis rather than systematic abnormal events.The implications of the overall pattern of megasporogenesis for the formation of 2n-eggs and the attempts to induce diplosporic or aposporic apomixis in potato are discussed.     

Ovule dichotomy per locule of the trilocular pistil in cassava,Manihot esculenta: a useful abnormality for increased seed production?

Histologically, the cassava pistil is trilocular with a single ovule in each locule producing a maximum of three seeds in nature. Heat stress (35°/45°C, night/day temperatures), as a physical treatment was attempted to induce useful reproductive and vegetative traits in 10 F₁ hybrids derived from ‘OMR 36‐41’and ‘OMR 36‐42’crosses, and two Asian cassava cultivars,‘Rayong 60’and ‘Rayong 1′, under greenhouse and field conditions. Cytoembryological examination of the repro‐ ductive organs revealed morpho‐structural features of normal pistil in all clones except ‘Rayong 1’which exhibited reproductive variations under greenhouse culture. Genetically, an important variation was ovule dichotomy per locule of the trilocular pistil so that each pistil contained six instead of the usual three ovules and lacked any outer integument. These ovules possessed functional embryo sacs with varying nuclear number. A pair of the dichotomous ovules possessed two embryo sacs of apparent different megaspore mother cells. Vegetatively, F₁ hybrids were dwarf (0.50m) in their phenotype and flowered 1 month earlier than the 1.5‐2.0m tall cultivars under greenhouse conditions.     

The effect of ovule age on ovary-slice culture and ovule culture in intraspecific and interspecific crosses with Tulipa gesneriana L.

The efficiency of two embryo rescue techniques, direct ovule culture and ovary-slice culture followed by ovule culture, is studied in crosses with Tulipa gesneriana as maternal genotype. The germination percentages increased, in most cases, significantly with increasing ages of te ovary-slices and ovules at the start of the cultures. The low number of embryos recovered at early culture dates is caused by a higher rate of embryo abortion and by retarded embryo development. The germination percentages for ovary-slice culture followed by ovule culture was mostly comparable to direct ovule culture. Unique hybrids have been obtained from the crosses T. gesneriana × T. agenensis and T. gesneriana × T. praestans. This revised version was published online in July 2006 with corrections to the Cover Date.     

Genotypic variation and polymorphism of 2S albumins of sunflower

Summary An early postfertilization ovule culture technique in Alstroemeria was described in which young ovules were cultured in vitro two days after pollination. This resulted in the direct production of seedlings in a normally incongruous cross combination. A histological study of the in vitro cultured ovules revealed the development of the embryo inside the ovule in the absence of normal endosperm. The in vivo grown ovules showed a total breakdown of the developing seed within four weeks after pollination due to a postfertilization barrier. This ovule culture technique was applied in a diallel cross of seven Alstroemeria species, five from Chile and two from Brazil. The selfings and the reciprocal crosses between the Brazilian species A. inodora and A. brasiliensis were congruous and gave seed set. The cross A. inodora x A. pelegrina gave viable seeds. In all other 39 combinations no viable seeds were harvested. In those cross combinations both pre-and postfertilization barriers were found. After early ovule culture hybrid plants were obtained in 27 of the 39 incongruous species combinations of the diallel.Abbreviations DAP days after pollination - MS medium medium according to Murashige & Skoog (1962)     

Seasonal changes in the incidence of apomixis of diploid,triploid, and tetraploid plants of Paspalum cromyorrhizon

Summary Changes in incidence of apomixis were determined at different intervals of the flowering period in a highly sexual diploid cytotype of Paspalum cromyorrhizon Trin., a natural tetraploid cytotype of the same species, and in the triploid and tetraploid intraspecific hybrids that were produced by pollinating the 2x cytotype with pollen of the tetraploid. Reproductive behaviour was determined from observations of embryo sacs in mature ovules. Gametophytic apomixis in this species in characterized by aposporous embryo sacs which differ from normal sacs by their number, orientation inside the ovule, their dimensions and shape, and the absence of antipodal cells.The potential for apomictic reproduction increased in relation to the ploidy level, suggesting a gene dosage effect in the incidence of apomixis. In the three ploidy levels, the highest incidence of apomixis was observed when the plants reached the maximum number of flowering heads. These findings suggest that the same environmental conditions that favour flowering should also be responsible for increasing the incidence of apomixis. An additional experiment under controlled conditions indicated that the light regime should be one of the factors that govern the amount of apomictic reproduction. Thus, polyploidy and apomixis should be of special significance in the dispersion and evolution of this grass species. The 4x facultative apomictic cytotype is far more frequent in nature than the highly sexual 2x. Moreover, most of the seeds produced in a flowering season should arise asexually. So, apomixis rapidly increases the number of plants bearing the successful maternal genotype, and sexual reproduction becomes important in adverse environmental conditions.     

Cultivar variation for seed development in white clover (Trifolium repens L.)

Summary A controlled environment study was undertaken to clarify the factors responsible for poor seed set and to study seed development, ovule degeneration and seed abortion, both morphologically and cytologically, in three Japanese cultivars of white clover. Although the mean number of ovules per floret was 4.2–5.1, the average number of seeds per floret was found to be only 2.3–2.7. Microscopic examination of carpels from 0 to 28 days following floret maturity and pollination showed that 26–33% and 8–17% of the total seeds lost occurred within the first three days and the third through fifth day following pollination, respectively. Beyond this period occasional seed abortion was observed at all stages of seed development, but this represented a very small proportion (2–7%) of the total seeds lost.A stain clearing technique was used to examine the cytoplasmic state of the embryo sac in intact, unfertilized, mature ovules and embryos of the ovules at 3 and 5-day periods following pollination. It was found that 20–22% of unfertilized and matured ovules were sterile, suggesting that ovule degeneration before fertilization was the major cause for the high percentage of seeds lost within a 0 to 3-day period following pollination. Cytological observations revealed that abortion of developing seed was due to a sudden arrest in embryo growth and that the early development of the embryo of such aborting seed was normal. Either nutrient shortage or meiotic irregularities may be the cause for high ovule sterility or post-fertilization abortion of developing seeds.     

An embryological study of a Pisum sativum x Vicia faba cross

Summary Flowers of Pisum sativum plants grown in the field and greenhouse were self-pollinated and pollinated with Vicia faba. Germination of pollen and pollen tube growth were slower in the Pisum x Vicia cross. About 2% fertile ovules in the field and 8% in the greenhouse were obtained from the Pisum x Vicia cross, compared to 26% and 48% for Pisum x Pisum. Development of the Pisum x Vicia embryo was normal in form, though much slower than the Pisum x Pisum embryo, and about the sixth day it began to collapse. The greatest difference between the two types of crosses was observed in the endosperm. While the Pisum x Pisum endosperm nuclei divided normally and endosperm developed throughout the ovule cavity and around the embryo, the Pisum x Vicia endosperm nuclei formed only scattered masses of densely stained nuclei which were not always in close proximity to the hybrid embryo. It was concluded that faillure of some ovules to develop following Pisum sativum x Vicia faba cross pollination was the result of slow germination of pollen and slow growth of the Vicia pollen tubes, with subsequent lack of fertilization. Collapse of fertilized ovules was associated with abnormal and limited development of the hybrid endosperm, possibly leading to lack of nourishment and eventual collapse of the hybrid embryo.     

谷子(Setaria italica Beauv)无融合生殖系SMA-1的发现

谷子无融合生殖的胚胎发生研究结果表明：谷子SMA-1的无融合生殖为无孢子生殖和二倍体孢子生殖两种类型。无孢子生殖类型与专性无融合生殖狼尾草相同，单核胚囊经二次有丝分裂形成四核胚囊，无反足细胞。二倍体孢子生殖类型与磨擦禾相似。胚珠发育不同时期的观察统计表明，该系前期的无融合生殖频率较高，达13.3%，在有性、无     

Interspecific hybridization between Vicia faba (L.) and Vicia narbonensis (L.): Early pod growth and embryo-sac development

Summary Fertilized embryo-sac development and pod growth was studied in one Vicia faba cultivar, one Vicia narbonensis population and their reciprocal crosses. The initial development of endosperm and embryo was at least four days faster in V. narbonensis than in V. faba. Pods and ovules developed also faster in V. narbonensis than in V. faba. The growth rate of the hybrid pods followed the growth rate of the mother species, but was slower than that of the pods from selfed flowers. In the cross V. narbonensis × V. faba the ovules stopped growing 9 days after pollination, while in the reciprocal cross they stopped growing 15 days after pollination. Hybrid embryo-sacs from V. faba × V. narbonensis were aborted before they reached the stage of 256 endosperm nuclei or 200 embryo cells. Selfed V. faba embryo-sacs reached this stage in less than 9 days after pollination. In the reciprocal cross the embryo-sacs were aborted before they reached the stage of 128 endosperm nuclei or 80 embryo cells. Selfed V. narbonensis embryo-sacs reached this stage at the 4th day after pollination. Given that at these stages the embryo has less than 200 cells it was concluded that an in-ovule embryo culture technique should be developed to obtain hybrid plants.     

Fertilisation assessment and postzygotic development in several intra- and interspecific Prunus hybrids

Summary the clearing technique was applied to assess fertilisation and to follow the postzygotic development in ten intra- and interspecific crosses among Prunus species used as rootstocks. The technique allowed to distinguish fertilised ovules from unfertilised ones, as well as to calculate the fertilisation percentage in intra-and interspecific compatible crosses. All the stages from unfertilised to fertilised ovules were analysed in order to determine the best time for embryo rescue. Differences in postzygotic development among the crosses are discussed in terms of the genotype of the seed parent in the pollen-pistil interactions.Abbreviations DAP Days After Pollination     

Hybridization between diploid (Gossypium arboreum) and tetraploid (Gossypium hirsutum) cotton through ovule culture

Summary With in vitro culture of ovules, interspecific hybrids have been obtained in an otherwise incompatible cross between a diploid (Gossypium arboreum) and a tetraploid (G. hirsutum) cultivated cotton. The early abortion of the embryo was prevented by repeated treatment of the flowers, immediately after pollination with a solution of gibberellic acid and naphthalene acetic acid. The ovules excised three days after pollination and cultured in a liquid medium underwent profuse proliferation, whereas on an agar-solidified medium supplemented with casein hydrolysate, indoleacetic acid and kinetin they germinated to form hybrid plants.     

The ovular incompatibility system in Gasteria verrucosa (Mill.) H. Duval

Summary The nature and process of ovular incompatibility in Gasteria verrucosa was studied. Using different crossing programs and following the pollen germination and ovule penetration histochemically, an ovular gametophitic incompatibility system based on two or more loci is affirmed. Pollen tubes after self-and cross-pollination penetrate the micropyle. Although the percentage of seed setting reached about 60% after cross pollination no clear differences could be seen between the germination and ovule penetration of cross and self pollen.In the ovules, multiple penetration of micropyles and a preference for the first position ovules was observed. Results of semi-compatible crossings and mixed crossings indicated a gametophytic two or more loci incompatibility system. Ovular incompatibility was manifested by the degeneration of the sporophytic ovular tissues. In Gasteria verrucosa the system of ovular incompatibility does not seem to be restricted to the ovule.     

High embryo sac fertility and diversity of abnormal embryo sacs detected in autotetraploid indica/japonica hybrids in rice by whole‐mount eosin B‐staining confocal laser scanning microscopy

An eosin B staining procedure for use with confocal microscopy (WE‐CLSM) was used to examine the mature embryo sacs in 24 typical autotetraploid (2n = 4x = 48) indica/japonica hybrids and to compare with their original diploid (2n = 2x = 24) indica/japonica hybrids in rice. Four of the 24 hybrids showed >80% embryo sac fertility, and 13 hybrids over 70%. The average embryo sac fertility of the 24 autotetraploid indica/japonica hybrids was 68.08%, which was 17.33% higher than that in diploid hybrids. When compared with the diploid hybrids, 79.17% of the autotetraploid hybrids had higher embryo sac fertility. A diversity of abnormal embryo sacs occurred in autotetraploid indica/japonica hybrids. Embryo sac abnormalities were similar for both autotetraploid and diploid hybrid, but their frequencies were different. Some novel abnormal types were found in autotetraploid hybrids, such as enlarged egg apparatus. The embryo sac fertilities and frequencies of various abnormalities in autotetraploid hybrids varied with parental genotype and environment. The average seed set of the autotetraploid hybrids were higher than their diploid hybrids in different growing seasons.     

The progamic phase and seed formation in Armoracia rusticana

The progamic phase is a period of specific interaction between the male gametophyte and the pistil tissues and is very important for proper seed development in plants. The aims of this study were to investigate the events during the progamic phase in Armoracia rusticana, to examine a plant’s capability of sexual reproduction and to obtain vital seeds under natural conditions. There were very few pollen grains on the stigmas at anthesis stage. The incompatibility reaction between the stigma cells and pollen grains was observed by using fluorescence microscopy. A series of ovary cross‐sections revealed a high accumulation of Periodic Acid Schiff (PAS)‐positive globules in all the embryo sacs. About 33% of ovules in the ovary were fertilized, but only 3% of them were able to germinate into seedlings.     

Embryo-rescue in the genus Tulipa L.; successful direct transfer of T. kaufmanniana Regel germplasm into T. gesneriana L.

Summary Embryo-rescue was studied as a means to overcome post-fertilization barriers in interspecific crosses in the genus Tulipa. With compatible T. gesneriana L. cultivar crosses, ovule culture was found to be superior to isolated embryo culture. Complete plantlet formation was possible from an embryo size of about 0.5 mm onwards.In the interspecific cross T. gesneriana x T. kaufmanniana, which is hampered by embryo breakdown, successful rescue of abortive embryos was demonstrated. Optimal embryo-rescue was achieved in cultures started seven to nine weeks after pollination. With cultures initiated at a later time, the rate of success decreased. A low number of germinative seeds were obtained after normal ripening of the seed pods, but by using ovule culture the efficiency of seedling formation could be increased dramatically.The ovule culture procedure will allow novel crosses and will offer new possibilities for the introduction of desirable genes into tulip cultivars.     

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     

Impact of heat stress on pod-based yield components in field pea (Pisum sativum L.)

Elevated temperatures associated with climate change result in crops being exposed to frequent spells of heat stress. Heat stress results in reduced yield in field pea (Pisum sativum L.); it is therefore important to identify cultivars with improved pod and seed retention under heat to mitigate this loss. Objectives were to investigate the effect of heat stress on phenology, yield and pod-based yield components. Sixteen pea cultivars were evaluated at normal and late (hot) seeding dates in the field in Arizona 2012 and in growth chambers with two temperature regimes (24/18°C and 35/18°C day/night temperature for 7 days) during reproductive development. We measured variation in the pattern of pod retention at four-node positions on plants, seed retention by ovule position (stylar, medial and basal) within pods and screened cultivars for pod retention, seed retention and yield. Heat stress reduced seed yield by accelerating the crop lifecycle and reducing pod number and seed size. Heat stress had the most damaging effect on younger reproductive growth (flowers and pods developed later), resulting in ovary abortion from developing flowers. Heat also accelerated seed abortion in all ovule positions within pods. Two high-yielding cultivars under control temperature, “Naparnyk” and “CDC Meadow”, maintained high yield in heat, and “MFR043” had the lowest yield. Cultivars “40-10” and “Naparnyk” retained the most ovules and seeds per pod, and “MFR043” aborted seeds when exposed to heat. In half of the cultivars, ovules at the basal peduncle end of pods were likely to abort while ovules at the medial and stylar end positions developed into seeds. For seven of the field cultivars, ovules at the medial pod position also produced mature seeds. Cultivars “40-10”, “Naparnyk” and “CDC Meadow” had greater pod and ovule retention or maintained high yield under heat stress, and were identified as heat-tolerant cultivars. Our results allow for a better understanding of pod-based yield components in field pea under heat stress and developing heat-tolerant cultivars.     

Doubled Haploids of Cocoa (Theobroma cacao L.) 1. Observations of Fertility

Ovule and pollen fertility have been studied in doubled haploids of cocoa derived from spontaneous haploids, doubled by colchicine treatment. The two measures of fertility were very variable; occasionally, the levels were as high as those of the parent plant, but most were lower. The low fertility was not improved either by changing pollinators or after grafting. Pollen fertility also varied seasonally more in doubled haploids than in their parents. The lack of differentiation of embryo sacs seemed to be the cause of low values of ovule fertility. Both of these unfavourable characteristics were not observed in the progeny of doubled haploids, The low fertility is described in terms of homozygote depression, forced, on normally allogamous plants, its consequences for the use of doubled haploids in the production of hybrid seeds are discussed.     

Regeneration and somaclonal variation in apomictic Paspalum dilatatum Poir

Summary In an attempt to incorporate variation into a uniform obligate apomict, plants of apomictic common dallisgrass, Paspalum dilatatum Poir., were regenerated from callus derived from immature inflorescences. Plants developed through both organogenesis and embryogenesis. A total of 682 regenerants were produced and more than 400 were transplanted into a field nursery and screened for somaclonal variation. Eventually 20 regenerants were selected, increased, and planted into a replicated nursery along with normal common dallisgrass. The characteristics examined were maturity date, plant height, number of racemes per inflorescence, number of spikelets per raceme, pubescence, stigma and anther color, ergot resistance, seed germination, seed set, pollen stainability, method of reproduction, and chromosome number. There were differences among the regenerants and between them and common dallisgrass for all traits except chromosome number, stigma and anther color, and ergot resistance. One of the more important regenerants had significantly higher seed set than common dallisgrass. All regenerants reproduced by aposporous apomixis but some exhibited a high degree of abortion while others had more aposporous embryo sacs per ovule than common dallisgrass. These findings demonstrate that common dallisgrass can be regenerated through tissue culture and that somaclonal variation is expressed in some of the regenerants, even though some of the altered traits are deleterious.