Pollen and pollination experiments. III. The viability of apple and pear pollen as affected by irradiation and storage

Summary Apple and pear pollen was irradiated with doses of 0, 50, 100, 250 and 500 krad (gamma rays) and stored at 4°C and 0–10% r.h. From the in-vitro germination percentages an average LD 50 dose of about 220 krad was estimated. For both irradiated and untreated pollen a close and corresponding lineair relationship existed between germination percentage and pollen tube growth.Irradiated pollen was much more sensitive to dry storage conditions than untreated pollen, resulting in less germination and more bursting. Apparently, irradiation caused the pollen cell membrane to lose its flexibility faster than normal. Rehydration of dry-stored, irradiated pollen in water-saturated air restored germination percentages up to their initial levels. The importance of this procedure in germination trials is stressed.     

Pollen and pollination experiments. I. the contribution of stray pollen to the seed set of depetalled,hand-pollinated flowers of apple

Summary Using the finger nails to remove most of the petals is a fast method of preparing apple flowers for hand pollination in the balloon stage. Such flowers still appeared quite attractive to insects; left to open pollination, the seed set per flower averaged two thirds of that of similarly treated, but hand-pollinated flowers. With the aid of marker pollen of scab or mildew resistant donors and the subsequent screening of the seedling progenies for resistance, it was shown that the stray pollen was responsible on average for one of every three seeds formed by unbagged depetalled and hand-pollinated flowers.     

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 and pollination experiments. V. An empirical basis for a mentor pollen effect observed on the growth of incompatible pollen tubes in pear

Summary The self-incompatible pear cultivar Doyenne du Comice was selfed with the aid of the mentor pollen technique (self pollen mixed 1:1 with compatible pollen) and the pioneer pollen method (compatible pollen applied 14 h in advance of the self pollen). Observations on tube growth in the style showed that inviable methylated pollen was ineffective either as mentor or pioncer pollen, having no effect on the performance of the self pollen which stopped growing at about one quarter of the style from the stigma. Calculations made on the basis of the obtained data indicated that the viable untreated or irradiated pioneer and mentor pollen, the former somewhat better than the latter, aided the self pollen tubes to reach the base of the style.     

Pollen and pollination experiments. VII. The effect of pollen treatment and application method on incompatibility and incongruity in Lilium

Summary Four methods were investigated for their capacity to overcome incompatibility and incongruity in Lilium longiflorum and the Lilium hybrid Enchantment. Temperature treatments of pollen at 40° to 60°C for one hour had no effect on germination capacity. None of the temperature treatments influenced incompatibility or incongruity. Application of cut-style pollination had a positive effect on self-pollination of Enchantment, but did not overcome self-incompatibility in L. longiflorum. With the aid of genetically inactivated (by irradiation) compatible mentor or pioneer pollen self-incompatibility of L. longiflorum could be largely overcome. Both methods also gave promising results in realizing interspecific crosses.     

Hybrid Tea-rose pollen. II. Inheritance of pollen viability

Summary Pollen viability was evaluated in about 500 seedlings originating from 31 crosses between nine commercial Hybrid Tea-rose varieties. The data indicated that pollen viability was inherited additively.     

Hybrid Tea-rose pollen. I. Germination and storage

Summary Germination and storage trials were carried out with pollen of several rose varieties. The pollen grains germinated well in a 15% sucrose solution with 40 ppm boric acid. Staining the pollen with a 0.1% tetrazolium solution and standardizing the degree of colour at which the pollen grains are counted as viable, provided a good viability estimate, simpler to carry out than in vitro germination. Germination capacity and staining ability of the pollen were greatly impeded-about halved-by dehydration during storage in desiccators at low humidity. This effect could be corrected by humidifying the pollen beforehand for about one hour, though this pre-treatment increased the percentage of germinated pollen grains more than the percentage stained. There was no difference between the two percentages in fresh or in deep-frozen pollen.Pollen stored at 1°C and high relative humidity soon lost its germination capacity: between 0 and 20% humidity a considerable proportion of the pollen remained viable for 9 months and longer. Storage for the same period in vacuum-sealed glass tubes at –24°C maintained viability as well or better and would probably prolong it further. Some of the cold-stored pollen induced a reasonable seed set after one year, a low seed set was obtained even after two years of storage at 1°C and low humidity.     

The interaction between compatible and self-incompatible pollen of apple and pear as influcenced by their ratio in the pollen cloud

Summary The production of self-seed was investigated in apple and pear with the aid of viable and compatible mentor/ pioneer pollen in relation to the proportion of self-imcompatible pollen present in the pollen cloud.Treatments consisted of mixtures of compatible and self-incompatible pollen at ratios of 1:1, 1:2 and 1:9 applied once or twice or followed by self-pollination. Selfing only, whether once or twice, produced virtually no fruits or seeds, while mixed pollinations did. Generally, fruit set tended to decrease and the self-seed to increase with increasing amounts of self pollen in the pollinations.The pioncer-pollen method (compatible in advance of self pollen) appeared more effective than the mentor pollen method (compatible and self pollen mixed). The use of viable instead of dead mentor/pioneer pollen causes competition for the ovules with the self pollen, but has the advantage that, in addition to some self-seed, other seeds are formed which, e.g. in apple, are necessary to keep the fruits on the tree until harvest.Fruit set was moderately reduced at ratios of compatible pollen to self pollen not exceeding 1:9 with pear and 1:5 with apple which constitute maximal ratios in practice as regards pollinator trees: trees of the main cultivar. In fruit orchards and probably in nature with other species, the interaction between compatible and self pollen may result in more self-seed, and so suggests that inbreeding played a greater part in the evolution of self-incompatible species, than the meager results of articial self-pollination imply.     

The performance of double or mixed pollinations with compatible and self-incompatible or incongruous pollen of pear and apple

Summary Six pear and five apple trials were carried out to ascertain the outcome of combinations of compatible pollen (C) with self (S) or incongruous pollen (I) as to the pollination index (PI=seeds/pollinated flower). The PI of the mixture C+I (1:5) was consistently depressed as compared to that of the control C. The results of the double pollinations S/C and I/C were affected by the temperature at pollination; their PI's at <15°C were twice as high as those at >15°C, being well above and below the PI of C in the former and latter case respectively. The opposite was true for the C/S combination, the PI of which increased with the pollination temperature; the PI of C/I did not differ much from the PI of C, irrespective of temperature. The conclusion was reached that the interaction previously and presently found between compatible and self-incompatible pollen also exists to a fair extent between compatible and incongruous pollen. However, in pear neither the mentor nor the pioneer pollen technique proved to aid its hybridization with apple, the formation of self seed was not observed either. In apple the production of apple × pear hybrids was likewise doubtful, but the double pollinations S/C and C/S formed 4–10% self seed.     

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.     

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.     

Chromosome association and pollen fertility in some interspecific hybrids of Lilium

Summary Meiotic observations in PMCs were made in eight kinds of interspecific hybrids in Lilium. Three hybrids, 6134-S × L. cernuum. L. regale × L. leichtlinii maximowiczii and L. auratum platyphyllum × L. henryi showed respectively the mean chromosome association frequencies of 1.4_II, 0.8_II and 1.5_II per cell at MI which were the lowest values hitherto reported in Lilium hybrids. In L. longiflorum × L. henryi, 33.3° of the cells had 1 or 2 multivalents of three to five chromosomes in addition to uni- and bivalents. 12° bivalents and 12 univalents were invariably observed in the triploid hybrid L. longiflorum × L. cernuum. The hybrids between the species belonging to the different sections of the genus generally showed high pollen sterility, with some exceptions. A more or less remote genomic homology was found between the different sections in Lilium.     

Embryo development and fruit-set in pear induced by untreated and irradiated pollen

Summary Flowers of the pear cultivars Conference, Doyenné du Comice and Gieser Wildeman were pollinated with untreated and 50-krad-irradiated pollen. Fruitlets were sectioned at regular intervals up to 6 weeks after pollination. Tubes of the irradiated pollen grew slower, but within two weeks nearly as many embryos were initiated as in the case of untreated pollen. Thereafter, the proportion of degenerated embryos increased, although the endosperm sometimes persisted. Six weeks after pollination only about 21% of the seeds induced by the irradiated pollen still contained normal embryos. At or before that time in-vitro culture of the immature seeds will be necessary because few embryos survive until harvest. The irradiated pollen stimulated a rather large parthenocarpic fruit-set, presumably associated with endosperm rather than embryo development during the first 4–6 weeks after pollination. The possibilities of haploid production are discussed.     

Pollination with gamma-irradiated pollen and development of fruits,seeds and parthenogenetic plants in apple

Summary Four apple (Malus X domestica) genotypes, Erovan, Golden Delicious, R1-49 and X6677, were pollinated with a marked pollen irradiated by -rays at doses ranging from 125 to 1000 Gy. Pollination with such irradiated pollen affected fruit set, seed number and seed contents, and induced the formation of parthenocarpic fruits and the development of parthenogenetic embryos. The immature embryos extracted from seeds. 2 and 3 months after pollination, were cultured in vitro and germinated after 2 months of cold treatment (3°C). Haploid plants were obtained in all 4 genotypes, after pollination with pollen irradiated at doses from 200 to 500 Gy. The optimum conditions for induction of apple haploids, by irradiated pollen approach, have been established.     

An assessment of thein vitro germination capacity of pollen grains of five tea hybrid rose cultivars

Summary Germination trials of pollen grains of the tea hybrid rose cultivars: Ferry Porche, Bronze Masterpiece, Queen Elizabeth, John F. Kennedy and Lady X, were carried outin vitro, to determine whether these cultivars are suitable as pollen donors in a breeding programme.Pollen grains of all five cultivars germinated poorly in a medium containing sucrose only. Addition of boric acid in the medium improved the germination percentages. The highest germinability was observed in Ferry Porche and Lady X on 15% sucrose+50 ppm boric acid, and 20% sucrose+100 ppm boric acid, respectively. Addition of calcium nitrate in the medium reduced germination percentages in all cultivars.The effect of pH of the medium on pollen grain germinability depended on the cultivar and the composition of the medium. On a medium free of boric acid, the pH values between 5.5 and 7.0 caused a significant increase in the germination percentages of pollen grains of Lady X and Queen Elizabeth. The addition of boric acid decreased the responsiveness of the pollen of all cultivars to the pH changes of the medium.The germination capacity of pollen grains of Lady X and Ferry Porche was not affected by incubation temperatures ranging between 20°C and 30°C.On the basis of the above results Lady X and Ferry Porche are considered the most suitable for pollen donors.     

The site of action of the crossability genes (Kr 1, Kr 2) between Triticum and Secale. I. Pollen germination,pollen tube growth,and number of pollen tubes

Summary Seven genotypes of common wheat (Triticum aestivum L.) were crossed with rye (Secale cereale L.) in order to find the site or sites of action of the crossability genes, Kr ₁ and Kr ₂, of wheat. The data obtained, by fluorescence microscopy, were compared to the controls (wheat x wheat). The results indicate that the crossability genes have little effect on pollen germination and on the time taken for the pollen tubes to reach the microphyle, irrespective of their crossabilities with rye. The number of pollen tubes reaching the microphyle is, however, affected by the Kr-genes, as high crossable genotypes have more pollen tubes than the low crossable ones. There was a high correlation between the mean number of pollen tubes at the micropyle with seed set, which also reflects the crossability. The Kr-genes seem to manifest themselves in the retardation and inhibition of pollen tube growth between the style base and the top of the embryo sac, where the effect is most distinct in the low crossable genotypes.     

Pollen germination,pollen tube growth and fertilization following self and interspecific pollination of Paspalum species

Summary Crossability between most Paspalum species is very low. This study was undertaken to identify the impediments to hybridization. Accessions of P. intermedium Munro. ex Morong, P. jurgensii Hackel and P. dilatatum Poir were self-pollinated and crossed with one anther. Paspalum intermedium is essentially self-sterile but P. jurgensii and P. dilatatum are highly self-fertile. Following pollination, pollen germination and tube growth were studied by observing the pollinated pistils with fluorescent microscopy. Examination of self-pollinated pistils revealed that the pollen germinated shortly after contacting the stigmas. Germination was over 80% for the P. intermedium and P. dilatatum accessions but only 57% for P. jurgensii. Pollen tubes grew to the micropyle within 45 minutes after pollination in P. dilatatum and 1 hour and 15 minutes in P. jurgensii. However, in the P. intermedium accessions most tubes did not grow beyond the stigma and very few penetrated the style and ovary. Apparently stylar-incompatibility is the reason for the low selfed seed set. In the cross-pollinations, pollen germinated shortly after pollination and germination ranged from 57 to 88% for the different crosses. In all crosses the pollen tubes grew to the micropyle within 30 minutes to 2 hours after pollination indicating that a cross-incompatibility system is not the cause for low crossability among these species. By examining embryo sacs from P. intermedium × P. dilatatum, its reciprocal and P. intermedium × P. urvillei crosses, it was determined that gametes failed to unite in some crosses and this is a major reason for low crossability.     

Cytoplasmic male-sterility and source of pollen influence the expression of resistance to sorghum midge, Stenodiplosis sorghicola

Sorghum midge, Stenodiplosis (Contarinia) sorghicola (Coquillett), is an important pest of grain sorghum, and host plant resistance is an important aspect of control of this pest. This research investigated how cytoplasmic male-sterility and source of pollen influence the expression of resistance to sorghum midge. Sorghum midge emergence was significantly lower in panicles of midge-resistant and midge-susceptible cytoplasmic male-sterile lines when pollinated with AF 28 - a midge-resistant restorer line, than those pollinated with Swarna - a midge susceptible restorer line, indicating the presence of xenia effects. Maintainer lines (B-lines) of midge-resistant parents had significantly lower numbers of eggs and larvae than the B-lines of midge-susceptible parents. Male-sterile lines of the both midge-resistant and midge-susceptible lines were equally susceptible, indicating that resistance to sorghum midge is influenced by factors in the cytoplasm of the B-line. These findings will have an important bearing on the production of hybrids with resistance to insects. This revised version was published online in August 2006 with corrections to the Cover Date.     

Cryopreservation of pollen from two rose cultivars

Summary An investigation was undertaken on the storage characteristics of pollen collected from two English rose cultivars. A rapid decline in viability was observed in pollen stored at +4° C and –20° C, whereas the viability of pollen, stored at ultra-low temperature (–196° C), remained constant. Cryopreserved pollen was shown to retain its ability for fertilisation. The effects of the stage of flower development and anther dehiscence were assessed on both pre-and post-cryopreservation viabilities. Successful long-term storage of pollen will facilitate hybridisation of rose species and cultivars that do not flower synchronously.     

A comparison of apple and pear seedlings with reference to the juvenile period. II. Mode of inheritance

Summary Analysis of two incomplete half-diallel schemes of crosses, involving 22 apple and 33 pear progenies with 2500 and 5400 seedlings respectively, showed a highly significant GCA and an insignificant SCA variance for the juvenile period (J.P.). This indicates that the inheritance of the J.P. is of an additive nature, a mode of inheritance which is a function of the inheritance of a complex of factors governing growth. The mean J.P. of apple progenies varied between 3.4 and 5.0 years, that of pear progenies between 4.7 and 7.0 years. The implications for breeding are discussed.Also presented at the Juvenility Symposium held at West-Berlin, November 1976.