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.     

Effect of floral morphology on pollination in Brassica rapa L

Summary Flower structure, especially the anther–stigma separation (ASS), is well known to affect pollination efficiency, and thus to potentially increase or decrease seed production in crops. Therefore, investigating the relationship between flower characteristics and pollination ability is crucial to a full understanding of mechanisms to improve F₁ seed production in Brassica rapa. We used image analysis to measure three flower characteristics: short stamen height (SSH); long stamen height (LSH); and pistil height (PH) in seven cultivars. We calculated the ratio of PH to LSH as an index of anther–stigma separation (ASS). We investigated the number of pollen grains (NPG) deposited on the stigma and the seed-set percentage (SSP) under open-pollination and self-pollination conditions (with and without insects, respectively). Nested ANOVA indicated significant differences between the seven cultivars in the floral characteristics except for PH. Moreover, much larger variation was observed in NPG and SSP than in floral characteristics. Although stepwise multiple regression analysis indicated that plants with relatively high PHs produced more seed under self-pollination, the number of seeds that resulted from self-pollination did not affect seed production because of incompatibility. Therefore, the effect of the spatial position of pistils on the F₁ seed production was low. Possibly other factors such as the total pollen production and visiting times of pollinators were important factors in the low yields observed in some cultivars     

Study of morpho-physiological, phenological and reproductive behaviour in protogynous lines of Indian mustard [Brassica juncea (L.)]

Understanding the relationship between floral and reproductive traits are critical to understand the evolution of plant species in brassica and need for selecting lines as parental lines in hybrid seed prodcution. The protogyny nature of Brassica species are characterized based on the variations in floral morphology, protogyny interval, stigma receptivity, pollen biology, compatibility and seeds set. The variation in stigma morphology features (stigma exsertion prior to anthesis) and protogyny interval (ranged from 8 to 10 days) formed phenotypically distinct. Studies on stigma receptivity and pollen viability revealed significant variability and found to be at its peak up to 3 days after anthesis and reduced drastically thereafter. Compatibility crosses and post pollination events such as pollen deposition on the surface of the stigma and culminating with the entry of the pollen tube into the embryo sac, was rapid and completed within 2 days after pollination, whereas in in-compatible cross not even single pollen was germinated on stigmatic surface and consequently no pollen tube growth even after sixth day of pollination. In compatible crosses average seed set ranged from 3 to 8 seeds per pod from cross pollination and no seed set in self. The investigation revealed that variation exists for protogyny interval, stigma receptivity and compatibility in the protogynous lines of Indian mustard and this can be exploited for the production of hybrids without emasculation.     

Results of multiple pollination in blueberry (Vaccinium elliottii Chapm.)

Blueberry (Vaccinium section Cyanococcus, family Ericaceae) is a shrub that produces multiple-seeded berries in which only a fraction of the ovules develop into viable seeds. A recessive yellowleaf marker gene was used to evaluate the efectiveness of a single pollination versus multiple pollinations in producing seeds in Vaccinium elliottii Chapm. A multiple-pollination experiment was conducted to see if the first pollen applied or the second produced more progeny, and to see if multiple pollinations increased the number of seedlings produced. Flowers of diploid yellowleaf Vaccinium elliottii were pollinated from one to four times at daily intervals. Pollen from redleaf and yellowleaf plants was used to produce two types of seedlings that could be distinguished visually. Generally, percent berry set, seeds per berry, and seed germination were not increased by multiple pollination. After multiple pollination, the pollen applied first always produced the most seedlings. Prior pollination greatly reduced seed set from subsequent pollination, but subsequent pollination caused little or no decrease in the number of seeds set by the first pollination. This revised version was published online in July 2006 with corrections to the Cover Date.     

The inheritance of partial self-compatibility in Brassica oleracea L.: Results from a half diallel homozygous for a moderately recessive S-allele

Summary In a study of partial self-compatibility in Brassica oleracea, flower number, seeded siliqua and seed production were recorded on self-and cross-pollinated inflorescences of the progenies of a half diallel between six in bred Brussels sprout plants homozygous for the same moderately recessive incompatibility allele S₄₅.On both self-and cross-pollinated inflorescences significant amounts of additively controlled genetic variation were found for seed set per flower. For cross-pollinated inflorescences this was also the case for the two components of seed set, seeded siliquae per flower and seeds per seeded siliquae, but for self-pollinated ones only seeded siliquae production showed significant additive variation. Considerable heterosis and gene interaction were always present and a simple additive dominance model did not explain the variation.Two of the parents transmitted lower levels of partial self-compatibility to their progenies and, in one of these, dominant genes appeared to be responsible. The most important feature determining the production of self seeds was found to be the number of flowering sites at which the incompatibility mechanism failed rather than the number of seeds produced at each site.     

Production of interspecific hybrids between Alstroemeria pelegrina L. var. rosea and A. magenta Bayer by ovule culture

Interspecific hybrids were efficiently produced in the cross-incompatible combination between Alstroemeria pelegrina L. var. rosea and A. magenta Bayer by culturing immature ovules with placenta 7–14 days after pollination on 2 g/l Gelrite-solidified MS medium containing 3% (w/v)sucrose. The plants showed intermediate characteristics between the parents and their hybridity was confirmed by karyotype and DNA analyses. The mean number of chromosome association per PMC at metaphase I was 2.60I+6.70II, pollen stainability was20.8%, and they produced viable seeds after self-pollination. Furthermore, mature plants were obtained when the hybrids were backcrossed as male parents with both the parents. The backcross-progeny from A. pelegrina var. rosea × hybrids exhibited 3.8 to 79.7% pollen stainability and that from A. magenta × hybrids 78.8 to 98.3%. Almost all of these plants produced viable seeds after self-pollination, which implies that they can beutilized for breeding of novel cultivars of Alstroemeria. This revised version was published online in July 2006 with corrections to the Cover Date.     

Genotype environment interactions and sib content in F1 hybrid —brussels sprouts

Summary Sib content of F₁ hybrid seed lots obtained from crossing four highly self incompatible brussels sprout lines was examined at two locations for four years. Parental combinations accounted for by far the greatest proportion of the total variation observed. Significant second and third order interactions terms suggest complex interactions between parental genotype, years, and locations. Attempts to interpret these data in terms of the regression model of Finlay & Wilkenson (1963), were unsuccessful.     

Investigations into the breeding system of cauliflower Brassica oleracea var. botrytis (L.)

Incompatibility reactions of 21 varieties of summer, autumn and winter type cauli-flowers were assessed by

1. (a)
   comparing the seed sets per siliqua from separate inflorescences of the same plant which were self-pollinated and mixed-pollinated with self and Brussels sprout pollen,     
   
   

Increased atmospheric humidity post pollination: A possible aid to the production of inbred line seed from mature flowers in the Brussels sprout (Brassica oleracea var. Gemmifera)

Summary In two highly self incompatible inbred lines of Brussels sprouts the effect of increased atmospheric humidity post pollination was examined immediately following 1) hand pollination of green buds and open flowers, and 2) blowfly pollination of open flowers. Data were obtained for mean number of seeds set per pollination, mean number of fruits setting seed, and mean number of seeds produced per fruit which set for both varieties. Measured as number of seeds produced per minute spent pollinating, it was clear that open flower pollination followed by high humidity conditions was a much more efficient method of producing inbred line seed (46 seeds/minute) than green bud pollination (27 seeds/minute).     

The effects of bud self-pollination and open flower self-pollination on the field characteristics of broccoli (Brassica oleracea var. Italica)

Summary Broccoli (Brassica oleracea L. var. italica) progenies grown from seed obtained by greenhouse budpollination (BP), open-flower self-pollination (OS), open-flower self-pollination with ether treatment of the stigma (OSE), and cross.pollination (CP) were compared in the field. For plant height, plant weight, head diameter, and head weight, BP resulted in the lowest values, OS and OSE were intermediate, and CP gave highest values. BP was highest, OS and OSE intermediate, and CP lowest for days to head emergence and days to harvest. Differences between OS and OSE were slight and all were non-significant for the 7 lines with complete data, but were significant for plant weight and head diameter when 20 lines were analyzed by the unweighted means method. The comparison BP versus OS-OSE was non-significant for plant height in both analyses and for head weight in the 7-line analysis only. In a second experiment using only BP and OS, and omitting days to head emergence, trends were similar but only the effects on head diameter were statistically significant. In the greenhouse, BP produced much more seed than either OS or OSE, and CP produced more than BP. Treatment of the stigma of open flowers with ether to overcome self-incompatibility did not result in greater seed production. Weight/100 seeds was higher in CP but did not differ among the three self-pollination methods.Technical Paper 5089, Oregon Agricultural Expriment Station.     

Pollen germination and pollen tube growth following self-pollination and intra- and interspecific pollination of Medicago species

Summary Pollen germination and pollen tube growth after self- and intra- and interspecific pollinations of annual [M. scutellata (L.) Mill., M. disciformis DC., and M. rigidula (L.) All.] and perennial (M. sativa L.) Medicago species were studied using a Carl Zeiss microscope with a fluorescence filter attachment.Self-pollination of self-fertile annual and intraspecific pollination of cross-fertile perennials were characterized by high pollen germination and rapid normal pollen tube growth. In contrast pollen germination percentages were very low and many pollen tube growth abnormalities occurred following interspecific pollination and self-pollination of self-sterile plants. The time period from pollination to fertilization for interspecific crosses was about double that following self-pollination of the annual species. However, fertilization occurred frequently following interspecific pollinations. Much of the sterility was due to factor(s) operating after fertilization.Cooperative investigations of the Kansas Agricultural Experiment Station, Dept. of Agronomy and the U.S. Department of Agriculture, Agricultural Research Service. Contribution No. 82-582-J- Mention of a trademark name or proprietary product does not constitute a guarantee or warranty by the U.S. Department of Agriculture, and does not imply its approval over other products that also may be suitable.     

Overcoming the self-incompatibility of Lilium longiflorum Thunb. by application of flower-organ extract or temperature treatment of pollen

Summary The application of flower organ extracts to stigmas and the temperature treatment of pollen were tried to overcome self-incompatibility of Lilium longiflorum cv. Georgia.Substances in stigma, style, ovary and anther were extracted with ethanol and fractionated with ethylacetate into the acidic, basic and aqueous fractions. The extracs melted in a small volume of distilled water were applied to stigmas prior to self-pollination. Hinomoto stigma extract, self-pollinated and cross-pollinated Georgia stigma extracts of high concentrations and Georgia anther extract of high concentration were effective in overcoming the self-incompatibility and resulted in a high percentage of fruit set and many normal seeds. Extracts from Hinomoto ovary, style and anther were ineffective, except a basic fraction, which was very slightly effective.Pollen was treated with 40°C for 60 or 90 minutes and 50°C for 30 or 60 minutes, and a half of each lot was followed by –20°C for 24 h, prior to self-pollination. All treatments were effective, especially at 40°C for 60 minutes or 50°C for 30 minutes, and 40°C for 90 minutes or 50°C for 60 minutes followed by –20°C for 24 h.     

Inter-accession variation in floral morphology in radish (Raphanus sativus L.)

We investigated variation in floral morphology among 36 accessions of radish (Raphanus sativus L.). We found wide, continuous inter-accession variation in 15 traits related to the size of the floral organs and the stigma position relative to the anthers of long stamens. A principal-component analysis of the 15 floral traits also showed wide variation among accessions. A large proportion of this variation was accounted for by lengths of stamens and the petal tube. In addition, the phenotypic stabilities with respect to the macro-environment (as affected by different sowing dates) and the micro-environment (among the flowers within a plant) differed between traits. Correlation analyses using all traits suggest the existence of genetic linkages or pleiotropies among floral organs. Our results indicate that floral morphologies varied greatly among radish accessions and they could be affected by the macro- and micro-environment. To explore the relationship between floral morphology and pollination efficiency, we compared the number of pollen grains deposited on the stigmas in the presence and absence of insect pollinators using three accessions with very different floral morphology. Because an accession with a higher stigma position than anthers might avoid self-pollination and an accession with a large stigma size received more pollen grains, floral morphology may affect pollination efficiency. Because the effect of pollination efficiency on seed production is well known, our results suggest that radish floral morphology affects seed production through its effect on pollination efficiency.     

A procedure suitable for in vitro pollen selection inBrassica oleracea

Summary Experiments to determine whetherBrassica oleracea pollen could effect fertilisation following incubation in liquid culture medium are reported. Pollen was incubated in vitro, collected by centrifugation and used to pollinate compatible pistils. While retaining viability and the capacity to germinate such pollen was unable to penetrate the stigma papillae. However, incubated pollen produced functional tubes following pollination of styles from which the stigmas had been removed. These tubes grew through the style to the ovary and viable seed was obtained. The potential application of this procedure in pollen selection is discussed.     

Cytological and genetical studies of a male sterile celery

Summary The inheritance and nature fo male sterility of a cerley (Apium graveolens L.) strain (MS1) is reported. Male sterility in MS1 is determined by a a recessive genotype for a single locus. Tests for linkage with the isozyme chromosome markers SDH-1 and PGM-1 were negative. MS1 male sterility was associated with a defective tapetum characterized by prominent vacuoles and premature degeneration. The stamens in the male sterile strain persisted in the flowers up to stigma receptivity, while in the male fertile plants they dropped before style expansion. The male sterile flowers produced normal amounts of nectar, resulting in cross pollination by various species of pollinators. It was estimated that the sterile strain produced 30% less seed than normal male fertile. Its possible use for hybrid celery seed production is discussed.Research supported by grants from the California Celery Research Board and BARD I-483-82.     

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.     

Molecular markers for yield components in Brassica juncea – do these assist in breeding for high seed yield?

A population of 112 F₁-derived doubled haploid lines was produced from a reciprocal cross of Brassica juncea. The parents differed for seed quality, seed color and many agronomic traits. A detailed RFLP linkage map of this population, comprising 316 loci, had been constructed, and was used to map quantitative trait loci (QTL) for seed yield and yield components, viz. siliqua length, number of seeds per siliqua, number of siliques per main raceme and 1000-seed weight. Stable and significant QTLs were identified for all these yield components except seed yield. For yield components, a selection index based on combined phenotypic and molecular data (QTL effects) could double up the efficiency of selection compared to the expected genetic advance by phenotypic selection. Selection indices for high seed yield, based on the phenotypic data of yield and yield components, could only improve the efficiency of selection by 4% of the genetic advance that can be expected from direct phenotypic selection for yield alone. Inclusion of molecular data together with the phenotypic data of yield components in the selection indices did not improve the efficiency of selection for higher seed yield. This is probably due to often negative relationships among the yield components. Most of the QTLs for yield components were compensating each other, probably due to linkage, pleiotropy or developmentally induced relationships among them. The breeding strategy for B. juncea and challenges to marker assisted selection are discussed.     

Introgression of unique characteristics of floral initiation under 24 hour day-length of Fragaria chiloensis ‘CHI-24-1’ into F. × ananassa

A unique flower initiation of the wild Junebearing strawberry strain ‘CHI-24-1’ in Fragaria chiloensis occurs under 24 h DL and high temperature conditions. To introduce the floral initiation characteristics of ‘CHI-24-1’ into cultivated strawberries of F. × ananassa, a cross pollination was conducted between ‘CHI-24-1’ and the Japanese short-day type strawberry cultivar ‘Nyoho’. The floral initiation of ‘CHI-24-1’ was induced in both parent and daughter plants linked with runners under a 24 h DL and 23/20 ^∘C, but not 8 and 16 h DLs at the same temperatures. Of the 21 F1 hybrids grown under the 24 h DL, 12 showed flower truss production in the parent and/or daughter plants linked with runners. Among 64 F1 hybrids, 26 exposed to 8, 16 and 24 h DLs for 30 days produced no flower trusses. However, 32, one and five F1 hybrids produced flower trusses under the 24 h DL alone, 8 h DL alone and both the 8 and 24 h DLs, respectively. The results of the experiments indicated that none of the F1 hybrids were day-neutral plants, but approximately 60% had the characteristics of floral initiation under 24 h DL, which was inherited from the pollen parent of ‘CHI-24-1’. The importance of the unique floral initiation characteristics under 24 h DL with high temperature for strawberry breeding was discussed.     

The effect of temperature on seed set and embryo development in reciprocal crosses of wheat and barley

Summary The effect of temperature on seed set and embryo development in reciprocal crosses of barley and wheat was assessed in crosses involving two spring barley varieties (Betzes, Martonvásári 50) and one wheat variety (Chinese Spring). Detached tillers placed in nutrient solution were pollinated in controlled environments at constant day-night temperature regimes (12, 15, 18 and 21° C) with a light intensity of 30,000 lux and a relative humidity of 80%. When barley was used as the female, lower temperatures (12 and 15° C) produced the maximum seed set, whereas for the reciprocal cross, the highest temperature (21° C) produced the best seed set in the Chinese Spring × Betzes combination.Low temperature retarded the embryo development. The highest numbers of hybrid plants were produced at 18° C and 21° C in the barley × wheat cross and in the wheat × barley cross, respectively. Embryos of about 1.5 mm length in the barley × wheat cross, and of about 1.0 mm length in the wheat × barley cross germinated successfully. The smallest embryo giving rise to hybrid plants was 0.57 mm in the barley × wheat cross and 0.51 mm in the wheat × barley cross.     

Interspecific hybrids of Cyclamen persicum and C. graecum

Summary Interspecific crosses were made to introduce the disease resistance of Cyclamen graecum into C. persicum cultivars and the abortive hybrid embryos were rescued by ovule culture. Diploid and tetraploid cultivars of C. persicum (CPD, 2n=2x=48; CPT, 2n=4x=96) were the pistillate parents and wild form of C. graecum (CG, 2n=84) were the staminate parents. After pollination, crossed ovaries were periodically collected and examined using paraffin sections. Histological observations suggested that both hybrid ovules of CPD × CG and CPT × CG should be transferred to the culture medium 35 days after pollination. Based upon this observation, crossed ovaries were collected 35 days after pollination, then ovules with placenta were explanted on culture medium and cultured in the dark at 25°C. Plantlets were induced from ovules cultured in MS medium containing 3% sucrose and 10% coconut milk. The hybrids (2n=66) derived from CPD × CG failed to yield viable seeds by self-pollination, although they showed some pollen fertility. In contrast, the hybrids (2n=90) derived from CPT × CG showed high pollen fertility and yielded viable seeds by self-pollination. Furthermore, they were resistant to disease caused by Fusarium oxysporum f. sp. cyclaminis, Erwinia herbicola pv. cyclamenae and Pseudomonas marginalis pv. marginalis.Abbreviations CPD C. persicum diploid - CPT C. persicum tetraploid - CG C. graecum