Unilateral cross compatibility in Vaccinium elliottii × V. arboreum, an intersectional blueberry hybrid

Vaccinium elliottii Chapmn., a diploid blueberry in Vacciniumsection Cyanococcus, was crossed with V. arboreum Marsh, a diploid blueberry insection Batodendron. The goal was toproduce hybrids that could be used toincorporate traits from these species intotetraploid southern highbush cultivars. The crosses were made reciprocally usingtwo clones of each species. A recessiveseedling marker gene for anthocyanin-freecotyledons in the V. elliottii clonesused as female parents permitted thetesting of mentor pollination. For thementor pollinations, V. elliottiiplants that were homozygous for therecessive marker allele were pollinatedwith a small amount of viable V.elliottii pollen carrying the recessivemarker mixed with a large amount of V.arboreum pollen carrying the dominantmarker. Thousands of intersectional hybridseedlings were obtained when V.elliottii was the seed parent, both withand without mentor pollination, but nohybrids were obtained from 2735 flowerspollinated in the reciprocal crosses. Theseeds that gave rise to hybrid embryos weresmaller than normal V. elliottiiseeds, but germinated well. Hybridseedlings grew slowly at first, buteventually produced some vigorous plants inthe field, although many plants remainedmuch smaller than normal seedlings of thetwo parental species. Some of the hybridsproduced numerous flowers during the 6years they were observed in the field, buteven with many diploid, tetraploid, andhexaploid section Cyanococcus blueberryplants growing nearby that could haveprovided pollen for their flowers, thehybrids produced no viable seed.     

The origin of polyploids via 2n gametes in Vaccinium section Cyanococcus

Summary The production of 2n pollen (pollen with the sporophytic chromosome number) was evaluated in 4x and 6x taxa of Vaccinium section Cyanococcus. Mean frequencies of 2n pollen producers were 17.1% and 8.3% in natural 4x and 6x populations, respectively. The frequency of 2n pollen producers in the 4x species ranged from 8.6% (V. angustifolium) to 23.8% (V. pallidum). Level of 2n pollen production was genotypically variable (1% to 37.4%). The widespread occurrence of 2n pollen in 2x, 4x and 6x taxa suggests that sexual polyploidization was widespread and responsible for the origin of the polyploid species found in this genus. The frequency of 2n pollen producers was not significantly different between the 4x species and their putative 2x ancestors. These results support the origin of 4x and 6x taxa as a consequence of sexual polyploidization. Polyploids derived from sexual polyploidization would be expected to have increased fitness and flexibility due to the mode of 2n pollen formation. In blueberry species the predominant mode of 2n pollen formation is genetically equivalent to a first division restitution mechanism (FDR). FDR 2n pollen transmits a high percentage of the heterozygosity and a large fraction of the epistasis from the 2x parent to the 4x offspring.     

Procedure for artificial cross pollination in clove (Syzygium aromaticum)

Summary A technique for artificial cross pollination in clove is described. This involves emasculating flowers two days before anthesis, and pollinating four days later. Following this procedure a seed set of approximately 25% can be expected.     

Use of Vaccinium octoploids to facilitate 4x-6x gene transfers

The use of an 8x Vaccinium produced through doubling a tetraploid hybrid with colchicine was studied as a means of facilitating gene exchange between V. corymbosum L. and V. ashei Reade. Analysis included meiotic observations and crossability studies of the 8x plant, as well as attempting to develop 6x breeding lines. Meiotic analysis revealed the presence of micronuclei in Telphase II products and polyspory in sporads. In the crossability studies five 4x highbush plants were pollinated with 8x o2 4x pollen. Data gathered included: % fruit set, average weight/fruit, average number of seeds/fruit, average number of seedlings/pollination, and number of 6x seedlings. Octoploid pollinations were significantly lower than 4x pollinations in all parameters. One 6x plant was produced from the 4x-8x crosses but was found to be mitotically unstable, having somatic cells with chromosome numbers ranging from 48 to 168.     

Pollen and pollination experiments. II. The influence of the first pollination on the effectiveness of the second one in apple

Summary Pollinating apple cultivars twice with compatible pollen at an interval of one or two days produced about twice as many seeds per pollinated flower as a single pollination. With the aid of scab- or mildew resistant marker pollen, it could be shown that the second pollen formed on average twice as many seeds as the first. The first pollen appears to pave the way-partly at its own cost-for the second and was therefore called pioneer pollen.     

Pollen tube growth and fertilization after different in vitro pollination techniques of Aechmea fasciata

The effect of cut style and placental pollination on fertilization efficiency was studied for compatible Aechmea fasciata plants. These alternative in vitro pollination techniques resulted in lower penetration rates of the ovules by pollen tubes in comparison to pollination on the stigma. An explanation was found in the intervention of the normal pollination process,through which less pollen-pistil interactions were built up. After cut style pollination the percentage of ovule penetration by pollen tubes increased when a longer style part was left at the ovary. Probably fewer factors that control pollen tube growth are present in the lower style part. Pollen germinated on the ovules after placental pollination but only rarely penetration of the micropyle by a pollen tube occurred. Activation of the ovary, induced by in vivo prepollination for 6 hours, and pollination two days after anthesis did not increase the fertilization percentage. Grafting a style with active growing pollen tubes to ovules on the placenta (placental grafted style pollination) resulted in a higher fertilization percentage. Pollen tube growth through the style was essential for pollen tube guidance to the ovules and penetration of the micropyle. This revised version was published online in August 2006 with corrections to the Cover Date.     

Controls over pollination and ovule fertilization in Vicia faba

Summary Variation in the frequencies of pollination and ovule fertilization was generated among faba bean (Vicia faba L.) genotypes in experiments which compared self-and open-pollination, natural and extended photoperiods, presence and absence of honey bees and a range of plant populations.There was a near-1:1 relationship between the frequencies of pollination of stigmas and the fertilization of at least one ovule within the pistil. Pollination was the major barrier to fertilization of the pistil, and ovules, since no mechanism was evident in the style to prevent pistil fertilization. The frequency with which single ovules were fertilized in pollinated pistils fell as their distance from the stigma increased, and averaged 83.5% overall.The frequency of pollination was most influenced by the level of autofertility possessed by the genotype, and the activity of pollinators. Provision of honey bees to plants in caged plots showed that high levels of pollination could be achieved under open-pollinated conditions, but this was also influenced by autofertility. Increasing plant population in the field had only a small effect on pollination, but extending photoperiod caused the levels and timing of self-pollination in three genotypes to vary.Large differences were found among genotypes in their early stigma receptivity, that is, the frequency of stigmas found to be pollinated at the erect standard stage, prior to the flower opening and being visited by bees. The implications of early stigma receptivity for producing new cultivars were discussed.     

The breeding system of rubber (Hevea brasiliensis): an evaluation of controlled hand pollination methods

Summary Controlled hand pollinated pistils of rubber were observed using fluorescence microscopy to assess the efficiency of the universally-employed method for the production of progeny for plant breeding. The controlled hand pollination method conducted in the morning resulted in the deposition of a mean of 15.6 pollen grains on the stigma, with no stray pollination observed. Over 36% of the pistils had the potential to set fruit.Pollinations conducted in the afternoon at the normal time of anthesis, had double the fruit set potential of morning pollinations as measured by penetration of ovules by pollen tubes. Pollinator efficiency also varied, with excessive damage to stigmas resulting in reduced pollen germination and tube growth. There were differences between clones in both female and male fertility, in the proportion of pistils with more than three carpels and in the production of small abnormal stigmas. There was no difference in pollen tube growth following self- or cross-pollination, indicating that the self-sterility mechanism of rubber operates in the ovary. Pollen could be stored for 5 days at 5°C and 75% RH with a 22% loss of fertility.     

Screening of melon (Cucumis melo L.) germplasm for multiple disease resistance

Summary Melon germplasm was screened for cucumber green mottle mosaic virus (CGMMV), powdery mildew (Sphaerotheca fuliginea), downy mildew (Pseudoperonospora cubensis) and Fusarium wilt (Fusarium oxysporum f. sp. melonis) resistance under artificial conditions except downy mildew for which screening was done under natural epiphytotic conditions. High level resistance to all the four diseases was not recorded in any of the collections tested. Nevertheless, ertheless, resistance to three diseases was located in three germplasm. Wild Cucumis species C. figarei exhibited absolute resistance to CGMMV and Fusarium wilt and high level resistance to downy mildew. Phoot or snapmelon (Cucumis melo var. momordica) — a non-dessert from of Indian origin—was highly resistant to downy mildew and resistant to CGMMV and medium resistant to Fusarium wilt. Iroquois was resistant to powdery mildew and medium resistant to downy mildew and CGMMV.     

Mode of pollination and seed setting in Egyptian clover

Summary The average cross-pollination in Egyptian clover under caged and uncaged conditions was 0.51% and 1.44% respectively. The low percentage of cross-pollination under caged conditions was due to the absence of honey bees, calm atmosphere and reduced photosynthesis whereas higher outcrossing under uncaged conditions was both by wind and honey bees activities. Per cent seed set was higher under uncaged conditions (53.0%) compared with that of seeds set under caged conditions (26.3%) and uncaged + hand tripping (52.3%). The honey bees have been found to increase seed set in Egyptian clover by tripping mechanism.     

Control pollination and pollen management in Sesbania sesban (L.) Merr.

Sesbania sesban was amiable to controlled cross pollination when the emasculation and pollination operations were performed in the morning. A medium of 10% sucrose solution was found to be optimal for in vitro germination of the pollen grains. The pollen is quite tolerant of orthodox storage conditions enabling controlled crosses to be performed among accessions that flower in differing seasons of the year.     

Floral morphology affects seed productivity through pollination efficiency in radish (<Emphasis Type="Italic">Raphanus sativus</Emphasis> L.)

To examine the effect of stigma position and size on seed productivity through pollination efficiency in radish, the numbers of self and cross pollen grains on the stigmas and the seed productivity under insect-pollination were compared among four bred lines. Lines with a small stigma or a high stigma relative to the anthers on long stamens tended to receive fewer self and total (self + cross) pollen grains but showed a higher cross-pollination percentage (ratio of cross pollen grains to total pollen grains on stigmas) than those with a large stigma or a lower stigma. Additionally, a higher cross-pollination percentage was associated with a higher outcrossing percentage. This result suggests that variations in the two stigma traits affect the outcrossing percentage via the cross-pollination percentage. Therefore, it should be possible to prevent loss of F₁ purity by selecting new parental lines with floral morphology that favors cross-pollination percentage.     

Polyploidy and sexual polyploidization in the genus Vaccinium

Genus Vaccinium,consisting of blueberries, cranberries, lingonberries and many related wild species, includes diploid, tetraploid, and hexaploid species. Most evidence indicates that the tetraploid species are autotetraploids, with non-preferential bivalent chromosome pairing. Although homoploid interspecific crosses usually produce numerous fertile hybrids if the parents are from the same section of the genus, inter sectional crosses at the diploid level normally produce no seedlings, weak seedlings, or seedlings that are very low in fertility. There is a strong but not complete triploid block within Vaccinium. Even with insections, tetraploid × diploid (and the reciprocal) crosses normally give only a few tetraploid and a few triploid hybrids. Hexaploid × diploid crosses within sections are very hard to make, and the few hybrids that have been obtained are pentaploid. The frequency of 2n gametes varies,both among genotypes within species and among species. Vaccinium pollen is normally shed in tetrads, and the frequency of large pollen grains shed in dyads can be used to estimate 2n gamete frequency. Cultivated blueberries occur at both the tetraploid and the hexaploid levels, and there are important genetic resources in the diploids. Unreduced gamete production has permitted transfer of genetic material from the diploid to the tetraploid level and from the diploids and tetraploids to the hexaploid level via triploid hybrids. Intersectional crosses can occasionally produce tetraploid Vaccinium hybrids that appear to behave as amphidiploids and have medium to high fertility. CommercialVaccinium cultivars are normally propagated by cuttings. Intersectional hybridization, chromosome doubling, and asexual propagation could permit the production of novel hybrid combinations with value as ornamentals or in fruit production. This revised version was published online in July 2006 with corrections to the Cover Date.     

Optimal pollination conditions for seed set after a self-pollination,an intraspecific cross and an interspecific cross of marrow-stem kale (Brassica oleracea var. acephala)

Summary The effects of different pollination techniques, with and without emasculation and delayed pollination, were investigated to find the conditions for maximum seed set after self-pollination and intraspecific and interspecific crosses of Brassica oleracea var. acephala. The results indicated that the pollination conditions achieving maximum seed set vary with the type of pollination. After controlled self-pollination, the best seed set occurs in bud 3 to bud 10. For the intraspecific cross, the youngest flower and the oldest bud produced the largest number of developed ovules but bud pollination was productive to bud 8. The yields from these two pollination types were best when the female parent was not emasculated. In the interspecific cross with B. campestris cv. Marco the best results came from the youngest flowers and the oldest buds subjected to the standard practice of pollinating directly after emasculation. Possible reasons for these effects are discussed.     

‘Thermally aided pollination’: A new method of breaking self-incompatibility in Brassica oleracea L.

Summary Application of heat during self-pollination of open flowers by means of an electric mini soldering iron resulted in seed set in self-incompatible Brassica oleracea varieties. Temperatures of 60, 70 and 80°C were tested. Compared with bud-pollination, in some cases the so-called thermally aided pollination (TAP) method, gave a considerably higher seed yield at 70 and 80°C. In view of the ease and rapidity of TAP it should be ascertained how far this method can replace bud-pollination in maintaining inbred lines. The possible mechanism behind the TAP response is briefly discussed.     

Inheritance of seedling colour in faba bean (Vicia faba L.)

Summary The inheritance of purple seedling colour was studied, in relation to the genetic control of flower colour. It was found that purple seedling colour is likely to be controlled by a single gene and that the trait is dominant over green seedling colour. White flowering prohibited the expression of the purple seedling colour, and is therefore thought to be epistatic.This character can be used to estimate rate of outcrossing in breeding programmes, as well as contribute to our knowledge of the biosynthesis of plant pigments and secondary metabolites such as tannins.     

Factors affecting haploid production in wheat using the Hordeum bulbosum system. 2. The effect of the timing of pollination

Summary The effect of manipulating the timing of pollination on the fertilization frequency in wheat × tetraploid Hordeum bulbosum hybridizations was investigated as a possible means of overcoming varietal incompatibility in this cross. Pre- and post-anthesis pollinations were made on previously identified highly crossable and poorly crossable wheat genotypes and the stage of development of the individual florets at pollination was expressed in days from anthesis units.In both crossable and non-crossable wheat genotypes, a significant negative linear relationship was observed between floret age and fertilization frequency. The magnitude of the response of early pollination was dependent on the number of alleles for non-crossability present at the Kr loci. The non-crossable cultivar Highbury, possessing more than one allele for incompatibility, showed a slight response. The greatest response was shown by the highly crossable cultivar Chinese Spring, possessing alleles for crossability at all Kr loci. The single chromosome substitution line, Chinese Spring (Hope 5B), possessing the most potent allele for non-crossability, Kr ₁, showed an intermediate response. The H. bulbosum genotype had no effect on the magnitude of this response, influencing only the mean fertilization frequency.     

Intra species variation in Atylosia scarabaeoides (L.) Benth., a wild relative of pigeonpea (Cajanus cajan (L.) Millsp.)

Summary Atylosia scarabaeoides (L.) Benth., a wild relative of pigeonpea, possesses several useful genes which can be utilized for pigeonpea improvement. In the present study, 33 accessions of A. scarabaeoides were evaluated at ICRISAT Center during the 1987 rainy season for variation in some useful traits to identify parents for inter-generic hybridization. A large variation was observed for leaf components, seed size, pod length, seeds/pod, days to flowering, seed protein, sulphur amino acids, resistance to cyst nematode, phytophthora blight, sterility mosaic, fusarium wilt, pod borer, pod fly, and pod wasp. Only four accessions were found to have more than 28% protein content. Methionine and cystine contents were marginally higher than in pigeonpea but the variation was not large enough to utilize them in the breeding program. In A. scarabaeoides. accessions resistant to fusarium wilt, phytophthora blight, sterility mosaic, and cyst nematode were detected. Compared to pigeonpea, the A. scarabaeoides accessions were less susceptible to lepidopteran borer and were immune to pod fly damage. Accessions ICPW 89 and ICPW 111 in short- (100–120 days), and ICPW 94 and ICPW 118 in medium-duration (140–180 days) were identified as potential parents for use in inter-generic hybridization.ICRISAT Journal Article No. 967