The yielding capacity of tetraploid sugar and fodder beets

Since the discovery of the use of colchicine in plant breeding, breeders have created tetraploid sugar and fodder beet. The results obtained were not very encouraging and therefore they soon proceeded to develop varieties containing triploids.At the Foundation for Agricultural Plant Breeding, Wageningen, polyploidy breeding was started in 1950. The initial material comprised two sugar beet varieties and five fodder beet varieties. By cytological examination of the aberrant individuals in the first and the second year after colchicine treatment, seed was obtained which produced a high percentage of tetraploids. In total 793 C₂ families were tested on trial fields and from these 32% appeared to be better than the original diploid material.The breeders interested in the material were provided with seed and stecklings of all the families in the C₁, C₂ or C₃ stage. With the distribution of this material the building up of tetraploid families at the Foundation for Agricultural Plant Breeding has been closed.     

Crosses between Beta intermedia bunge and B. vulgaris L.

Summary A tetraploid annual male sterile form of Beta vulgaris L. (2n=4x=36) was crossed with the wild beet species Beta intermedia Bunge (2n=36). The resulting F₁-plants were male sterile annuals being two or three times back-crossed to diploid and tetraploid sugar and fodder beets in the next years. Apart from tetraploid material (36 chromosomes) hexaploid (54 chromosomes) and a number of aneuploid plants developed.The results obtained justify the conclusion that, at a tetraploid level the material mostly propagates apomictically after the F₁ generation. The presence of penta-, hexa-, septa-and even octaploid plants might be explained by assuming that no meiosis has taken place in the crossing partners. Triploid plants are sometimes found in the progeny of hexaploid material and may presumably be considered haploids. Moreover some pentaploid plants were found in the progeny of the open pollinated F₁ which after two generations of bagging are still pentaploids although they produce no pollen. This is another clear indication of apomictic reproduction.The tetraploid generation from the cross between the hexaploid material and diploid sugar beets probably contains the best prospects for breeding.     

The creation of tetraploid beets

Summary In 1950, 1951 and 1952 germinated seed of various sugar beet and fodder beet strains were treated with colchicine, with the object of obtaining tetraploid plants. The plants were examined cytologically in their first and second year. Only stalks of the C₀ generation (the treated one) in which no diploid cells were found were kept for seed production.By raising the tetraploid C₁ plants in greenhouses the year that the seed has been harvested and by vernalizing the young plants seed production is made annual. For vernalization the plants are kept in a potato storage room cooled by outside air to a temperature of about 5°C and under a day length of 24 hours.The results of the cytological examination demonstrated that collecting seed from tetraploid stalks of the C₀ presents advantages. The C₁ had about 77% tetraploids.It should be noted that the determination of the chromosome numbers of polyploid material is preferable to measurements of stomata or pollen grains as the latter are not reliable.The investigations are being continued. Yield trials with tetraploid material have been made by D. Kloen.     

Studies on colchicine induced tetraploids of jute

Summary Colchicine induced tetraploid jute plants were shorter than their diploid counterparts. C₂ tetraploid plants became taller than C₁ plants. Leaves were larger in size and with higher breadth/length ratios compared to those of diploids. Stomata were bigger than those in diploid leaves but their frequency was less in tetraploid leaves. Tetraploid flowers were bigger than the diploids and bore a reduced number of stamens. Tetraploid pollen grains were larger but the percentage of sterility in them was more than that of the diploids. Fewer fruits with a reduced number of seeds were formed in tetraploid plants. They usually were shorter but wider than diploid fruits. Tetraploid seeds were larger and heavier. C₂ tetraploid plants bore more fruits with an increased number of seeds than C₁ plants. Breadth of individual fibres increased but length remained unchanged.The paper was read at the Eighth Pakistan Science Conference held at Dacca, 1956.     

Autotetraploidy in caraway (Carum carvi L.) for the increase of the aetheric oil content of the seed

Summary An autotetraploid population of caraway (Carum carvi L.) was produced by treating the growing points of young plants with a 0.2% colchicine solution. Selection on morphology, pollen size and pollen shape, followed by a selection on seed size proved to be an effective way to identify the mixoploid C₀ plants. Three percent of the treated plants produced a tetraploid progeny. Moderate selection in the C₁ and the C₂ population resulted in an improvement of the average seed-set to the amount of 50% of the diploid control, with a range of 21–78%. The 1000-kernel weight of the tetraploid material was 7 g, compared to 3 g in the diploid control. The oil content of the autotetraploid seed was increased with 35.6%; the increase in carvone amounted to 6.9%. The results of the experiment indicate that breeding work in autotetraploid caraway may result in a new cultivar.     

The transfer of triazine resistance from Brassica napus L. to B. oleracea L. III. First backcross to parental species

Summary Atrazine resistant Brassica napus × B. oleracea F₁ hybrids were backcrossed to both parental species. The backcrosses to B. napus produced seeds in both directions but results were much better when the F₁ hybrid was the pollen parent. Backcrosses to B. oleracea failed completely but BC₁s were rescued by embryo culture both from a tetraploid hybrid (2n = 4x = 37; A₁C₁CC) and sesquidiploid hybrids (2n = 3x = 8; A₁C₁C). Progeny of crosses between the tetraploid hybrid and B. oleracea had between 25 and 28 chromosomes. That of crosses between the sesquidiploid hybrid and B. oleracea had between 21 and 27. A few plants that had chromosome counts outside the expected range may have originated from either diploid parthenogenesis, unreduced gametes or spontaneous chromosome doubling during in vitro culture. Pollen stainability of the BC₁s ranged from 0% to 91.5%. All the BC₁s to B. oleracea were resistant to atrazine.     

An efficient method to separate tetraploid from diploid plants after colchicine treatment of turnips (Brassica campestris var. rapa)

To obtain a C₁-population with a high percentage of tetraploids after colchicine treatment of turnips, a selection in the C₀ can be used. A comparison is made between two criteria for this selection, viz. the average pollen grain size and the variation in pollen grain size.C₀-selection on the basis of the average pollen grain size of five pollen grains per plant reduced the number of diploids in the C₁ considerably. When the variation in pollen grain size of these five pollen grains is included as a selection-criterion the number of tetraploids in the C₁ increased.The two methods are discussed in the light of the working efficiency and the genetical variation in the tetraploid C₁-population. It is concluded that the character of the initial material should govern the choice of the selection criterion.     

The creation of tetraploid beets

Summary The purity of the tetraploid material may decrease through various causes.In the first place foreign monoploid pollen exerts a detrimental influence because it has a greater germination energy and a greater mobility than diploid pollen.A good impression of the purity can be obtained by cytologically investigating the C₂ of a few beets from an isolated plot. For this purpose seed is sown out in the greenhouse in winter and at least 25 plants of the C₂ of some beets per plot are investigated. By taking care that material of early flowering and late flowering beets is disposed of, a sufficiently reliable impression of the purity of the tetraploid progeny can be obtained.The result of the cytological checking which was carried out in the winter of 1952/53, was disappointing.That a single aberrant beet can exercise a detrimental influence was proved in the varietyAlpha when an aberrant plant was not rejected. The number of non-tetraploids of this beet was much greater, while in a neighbouring plant 30% triploids were present on investigation.It is important that cytological investigation of further generations of the tetraploid material be continued because otherwise the material will soon become very impure.Finally it should be mentioned that the influence of early bolters was insignificant because of a different flowering time and that the occurrence of break down could not be established.

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Samenvatting Door allerlei oorzaken kan de zuiverheid van het tetraploide C₁ materiaal achteruitgaan.Vreemd monoploid stuifmeel zal een nadelige invloed uitoefenen omdat dit een grotere kiemenergie en een grotere bewegelijkheid bezit diploid stuifmeel.Een goede indruk kan worden verkregen van de zuiverheid door de C₂ van enkele bieten cytologisch te onderzoeken. Te dien einde wordt in de winter zaad in de kas uitgezaaid en van iedere nakomelingschap worden minstens 25 bieten onderzocht. Door er verder voor te zorgen dat materiaal van een vroegbloeiende en een laatbloeiende biet aanwezig is, zal een voldoend betrouwbare inddruk worden verkregen van de zuiverheid der tetraploide nakomelingschappen.Het resultaat van de cytologische contrôle, die in the winter van 1952/1953 werd uitgevoerd, viel tegen. Dit is waarschijnlijk een gevolg van het feit dat er bij het opppotten van de tetraploide C₁ bieten enkele fouten zijn gemaakt, waardoor later diploiden op het veld werden uitgeplant. Het volgend jaar werden daarom de rijtjes C₁ bieten in de kas zodanig gedund, dat de bietenafstand enkele cm bedroeg. Bovendien werden uit voorzorg de afgezonderde C₁ bieten vóór de bloei nogmaals onderzocht en planten, waarin een afwijkend aantal chromosomen werd aangetroffen, werdentijdig verwijderd.Bovengenoemde voorzorgsmaatregelen hebben een gunstige invloed uitgeoefend, want bij cytologisch onderzoek in de winter 1953/1954 bleek dat het aantal niettetraploiden veel geringer was. Dat een enkele afwijkende biet een nadelige invloed kan uitoefenen bleek in het rasAlpha, waarin een afwijker niet werd verwijderd. Het aantal niet-tetraploiden van deze biet was veel hoger, terwijl in een onderzochte buurplant 30 % triploiden voorkwam.Het is van belang dat het cytologische onderzoek in de volgende generaties van het materiaal wordt voortgezet aangezien anders het materiaal onzuiver zal worden.

     

Reproductive diversity and strategies for breeding in St. John's wort (Hypericum perforatum L.)

St. John's wort (Hypericum perforatumL.) is a medicinal crop plant that reproduces by apomixis. To study the reproductive diversity within the initial breeding material, 656 plants from 92 accessions of the species were analysed by flow cytometric seed screening (FCSS) and compared to 66 plants of cv. ‘Topaz’ as a control. Among plants from the 92 accessions, 16 were obligate sexuals, nine were obligate apomicts and all remaining ones were facultative apomicts. Among controls, there were six obligate and 60 facultative apomicts. Tests of reproductive pathways indicated that, in the investigated collection, all apomictic plants were tetraploid and all sexual plants were diploid. A strategy for a cross breeding program in St. John's wort is illustrated by crossing one diploid obligately sexual mother plant with six tetraploid facultative apomicts. All 91 F₁ progeny obtained were triploid with 58% being obligate sexuals and 42% facultative apomicts. To quantify the different routes of seed formation within facultatively apomictic individuals, the reproductive pathways were reconstructed from about 30 single seeds from each of five tetraploid and two triploid plants using FCSS. The frequency of seed formed by apomixis ranged from 54% to 67% among four tetraploid genotypes. One triploid and two tetraploids formed 23% to 73% of the so-called B_III hybrids (fertilization of unreduced eggs) and one triploid was obligately sexual. The results show that while the mode of reproduction in H. perforatum is highly variable, tetraploid obligate apomicts as well as diploid and triploid obligate sexuals can be selected as starting material for breeding purposes. This revised version was published online in July 2006 with corrections to the Cover Date.     

Effects of Inbreeding and Genetic Variation on Forage Quality Traits and Dry Matter Yield in Dactylis glomerata L. Subspecies

Cultivated 4× clones, 2× germplasm and their respective S; progeny were utilized in this study. Dry matter yield and forage quality data (in vitro dry matter disappearance, cell wall constituents, total nitrogen) were obtained on composited vegetative plant material harvested during four successive 4-week growth periods in the greenhouse. There was no detectable average inbreeding effect for any of the traits. The group mean of the diploids was significantly lower for in vitro dry matter disappearance than the tetraploid group mean. However, two out of six diploid 51 families exceeded the highest tetraploid family for this trait. The diploids had significantly higher cell wall constituents than tetraploids except for acid detergent lignin. There was more variation for any trait at the diploid level than at the tetraploid level among S₁ families as well as within families. Dry matter yield and total nitrogen were significantly correlated in diploids and in tetraploids. However, little variation was detected lot total nitrogen at either ploidy level. The results suggest that selection for improved quality in progeny from interploidy crosses may be more efficient by first selecting at the diploid level followed by scaling to the tetraploid level, rather than selection at the tetraploid level after hybridization.     

The transfer of triazine resistance from Brassica napus L. to B. oleracea L. II. Morphology,fertility and cytology of the F1 hybrid

Summary F₁ hybrids of triazine resistant Brassica napus and triazine susceptible B. oleracea were morphologically intermediate to the parent species. Of 49 hybrids examined, 44 had 28 chromosomes, two had 37, one had 38 and two had 56. The 38-chromosome plant was thought to be a matromorph, the others, A₁C₁C (28), A₁C₁CC (37) or A₁A₁C₁C₁CC (56) type hybrids. Pollen stainability averaged 9.0% in the sesquidiploid, 32.0% in the tetraploids and 89.5% in the hexaploids. All the interspecific hybrids were resistant to 1.0×10^-4 mol L^-1 atrazine. The sesquidiploid hybrids produced gametes with chromosome numbers ranging from 9 to 17 and the tetraploid hybrid gametes had chromosome numbers from 15 to 22. Most hybrids produced self-seed. The partial fertility of these hybrids may permit their backcrossing to one or both parents.     

Introgression of Cajanus platycarpus genome into cultivated pigeonpea, C. cajan

Summary Cajanus platycarpus, an incompatible wild species from the tertiary gene pool of pigeonpea (C. cajan (L.) Millspaugh), has many desirable characteristics for the improvement of cultivated varieties. To necessitate such transfers, embryo rescue techniques were used to obtain F₁ hybrids. The F₁ hybrids were treated with colchicine to obtain tetraploid hybrids, that were selfed to obtain F₂, F₃ and F₄ progenies. All of the hybrids and subsequent progenies had an intermediate morphology between the two parents. Backcrossing of the tetraploid hybrids with cultivated pigeonpea was not possible given embryo abortion, with smaller aborted embryos than those obtained in the F₀ parental cross.As a route of introgression, diploid F₁ hybrids were backcrossed with cultivated pigeonpea and BC₁ progeny obtained by in vitro culture of aborting embryos. BC₂ plants were obtained by normal, mature seed germination. Although embryo rescue techniques had to be used to obtain F₁ and BC₁ plants, it was possible to produce BC₂ and subsequent generations through direct mature seed. Every backcross to cultivated pigeonpea increased pollen fertility and the formation of mature seeds.Special project assistant till December, 2003.     

Problems in breeding the legume Indigofera spicata for tropical pastures

A survey of I. spicata introductions showed the majority to be tetraploid perennials and similar to each other with red stems, strongly stoloniferous habit, and poor seeding ability. The rest were green stemmed annuals or biennials which were either diploid or tetraploid and which were free seeding and almost non-stoloniferous.Crosses were attempted using several green stemmed introductions as the female parents with a representative red stemmed one as the male. The ploidy of a diploid parent was raised with colchicine treatment before crossing. F₁ seed was obtained in all crosses except in that involving the induced tetraploid. The F₁ generations were sterile except for one cross from which a small quantity of F₂ seed was obtained. A high degree of sterility was maintained in the F₂ and F₃ generations but fertile F₃ selections produced mostly fertile progeny. The sterility could be followed by the percentage stained pollen and appeared to be controlled by genetic factors.Normal types of distribution of the phenotypic classes for yield and stoloniferous development were obtained for the F₃ population which contained 26.3 per cent perennial plants. Two F₃ lines were obtained which combined the desirable characters of the two parents.The effect of temperature and photoperiod on all introductions used as parents and the two promising lines was investigated in a phytotron. All types flowered at both the 8 hour and 16 hour photoperiods and temperatures 27°/22°C and 30°/25°C favoured flowering. The better flowering of the green stemmed parent had apparently been transferred to the selected F₃ lines. The highest plant dry weights occurred at 30°/25°C and the effect of photoperiod on yield was reversed as the temperature increased.     

The induction of tetraploidy in chicory (Cichorium intybus L. var. Magdebourg) by protoplast fusion

Summary Fusions of chicory leaf protoplasts were realized in order to obtain a higher number of tetraploid plants compared to the traditional doubling techniques with colchicine. Thanks to the fusion technique, using a mixture of PEG, DMSO and a solution of CaCl₂ (pH 10.5), 25% tetraploid (2n=36) plants were obtained. Among the 167 regenerated plants, only one aneuploid of a near tetraploid level (2n=33) was identified.     

Tetraploid Westerwolths ryegrass

Summary Germinated seeds of the annual Westerwolths ryegrass were treated with colchicine-solutions of 0.05 to 0.4% at a temperature of 29°C in order to obtain autotetraploids. The best results were obtained after treatment for 2 hours with 0.1 and 0.2% solutions or 4 hours with 0.05%. By these procedures about 8% of the treated seeds produced mixoploid or tetraploid mature plants. A preliminary progeny test was performed with seed of outstanding C₁-plants.In spaced plants and in drilled rows the dry matter production of these C₂-families was on average equally as high as that of the most productive original diploid variety. The proportion of leaves was higher in both the first and in the second cut, while drought resistance was considerably better. The tetraploids came later into ear, tillering ability was smaller and mildew attack was slightly heavier. In samples from drilled and from space-planted rows seed production per inflorescence was found to be higher in the tetraploids than in the diploids. The seed of the tetraploids was larger.From the data available it is concluded that chromosome doubling offers favourable possibilities for the improvement of Westerwolths ryegrass. Some suggestions for a breeding programme are given.     

Tetraploid Italian ryegrass (Lolium multiflorum Lam.)

1. 1.
   Some preliminary experiments are described with C₂- and C₃-generations of tetraploid Italian ryegrass. In all cases the tetraploids had a lower dry matter content than the diploids.     
   
   

Cross compatibility between diploid and tetraploid perennial ryegrass (Lolium perenne L.)

Summary A number of crosses were made between diploid and tetraploid Lolium perenne plants in order to determine the degree of cross compatibility between the two different ploidy levels. The range of hybridization undertaken involved diploidxdiploid, and tetraploid x tetraploid control crossings, diploid x tetraploid crosses, tetraploid x diploid crosses and mixed pollinations. The seed set, the germination capacity of resultant hybrid seeds, and the chromosome constitution of derived seedlings was determined. In addition attempts were made to culture 12 day-old hybrid embryos from the diploid x tetraploid and reciprocal crosses.The crossing results obtained indicated no barrier to fertilization between diploids and tetraploids, but the irregular and poor development of the seed accompanied by much reduced germination, indicated unsatisfactory endosperm establishment of large numbers of triploids, as confirmed by the result of culturing embryos dissected from 12-day old seeds.In preliminary studies undertaken to determine the extent to which diploid and tetraploid ryegrass varieties intercross when grown in close proximity for seed production, Griffiths and Pegler (1966) observed a very low incidence of triploids amongst the progenies of diploid plants exposed to the pollen of tetraploid plants and also amongst the progenies of tetraploid material exposed to the pollen of diploid plants.In these and subsequent studies it was noted that only a very small proportion of the so-called fully formed seeds derived from diploid x tetraploid crosses did actually germinate. When, in 1964, propagules of clones of S.24 perennial ryegrass, well separated from one another, were placed in drills of the tetraploid perennial ryegrass variety Reveille, the proportion of florets which formed caryopses, and of caryopses which germinated was as follows (Table 1).The data (Table 1) clearly indicate that a considerable proportion of the S.24 florets had been fertilized by 2n pollen and had formed caryopses, but most of these had proved to be defective. As noted in the previous studies, the frequency of triploid seedlings was low, thus suggesting incompatibility between the n and 2n gametes for the production of viable zygotes.Of the total progenies classed as tetraploids in the early seedling stage, 80% proved to be ryegrass x tall fescue F₁ hybrids. These arose as a result of free crossing with     

Dihaploid Plant Production from 4 × -Genotypes of Potato by the Use of Efficient Anther Plants Producing Tetraploid Strains (4 × EAPP-Clones) — Proposal of a Breeding Methodology

Tetraploid clones of potato with a superior efficiency in producing androgenetic plants (4 × EAPP-clones) have been obtained by culturing in vitro anthers of 2 × EAPP-clones. The latter were isolated by three cycles of recurrent selection from diploid breeding material (UHRIG 1985 a). In this paper we report on the capacity of 4 × EAPP-clones, when crossed to in vitro unresponsive 4 × genotypes, to transmit to their F₁ their androgenetic potential, Five 4 ×× 4 × F₁ crosses were considered, which produced on average 35 embryoids per flower – a value higher than that of nonresponsive 4 × genotypes (no embryoids obtained), but-also significantly better than the value found for 4× EAPP-clones (9.8 embryoids per flower). The hybrid families behaved differently from each other, with one producing up to 63 embryoids per flower. A range of per plant values was, moreover, found, revealing the existence of a large variability among sister plants belonging to the same F₁ cross. The presented data indicate a rather simple inheritance of dominant genetic factors acting in favor ot androgenesis. They also suggest that the utilized 4× EAPP-clones were possibly heterozygous for such genes. Data are also reported on the ploidy level of anther plants obtained from 4× and 2× EAPP-clones. In this respect 2× EAPP-clones show the interesting capacity of generating, via anther culture, a consistent fraction of tetraploid plants (13,7 %). Based on the findings reported in this paper we propose, for tetraploid S. tuherosum L., a cyclic breeding procedure making use of anther culture and where ploidy level alternates, within a cycle, between 2× and 4×.     

Colchicine mutagenesis is associated with substantial heritable variation in cotton

Summary Seedlings of the cotton cultivar Sicot 1 were treated with either 1%, 2% or 5% colchicine in lanolin which was topically applied to the apical meristem four days after emergence. Of 293 1%-treated seedlings 64 survived to produce fertile C₁ shoots which gave rise to 84 C₂ single boll families. C₂ and C₃ generations were examined in field experiments for morphological variants and for a number of quantitative characters (yield components and fibre quality). No morphological mutants were seen but considerable heritable variation for metrical characters of economic importance was detected. Of 18 extreme C₂ families consisting of 54 C₃ progenies, 19 were shown in a replicated spaced-plant trial to be significantly different to the Sicot 1 parental material for at least one (usually more) characters. The 222 C₃ progenies of the 66 non-extreme C₂ families were tested in an unreplicated plot experiment and at least 14 were suspected of exhibiting variation in metrical characters. Although no light has been shed on the mechanism responsible for the production of variants by this method, it is argued that the colchicine technique results in higher levels of genetic variation than found in commercial cultivars; variation which can be readily exploited by cotton breeders.     

Production of 2n pollen of Asiatic hybrid lilies by nitrous oxide treatment

Tetraploid varieties of lilies have superior agronomic traits such as large flowers and resistance to physiological disorders. In the present study, we attempted to induce 2n pollen of Asiatic hybrid lilies by arresting the meiotic process with nitrous oxide (N₂O) gas. To determine which meiotic stage is optimal for induction of 2n pollen, plants with attached buds at different meiotic stages were treated with N₂O for 24 h in a pressure-tolerant cylinder. A few 2n pollen grains were induced using plants with anthers in prophase I, whereas mixed pollen grains of differing size were produced using plants undergoing meiotic metaphase predominantly in anthers. Although normal lily pollen grains are elliptical, nitrous oxide exposure induced giant pollen grains that appeared spherical. Flow cytometry analysis showed that the giant pollen grains were diploid. When mixed pollen that included normal and giant pollen was crossed to tetraploid cultivars, the resulting seedlings were tetraploid and aneuploid, indicating that the giant pollen grains were diploids that could generate tetraploid seedlings through fusion to diploid eggs supplied from a tetraploid female parent. Thus, treatment with N₂O is useful for the production of 2n lily pollen and may provide a new approach for tetraploid lily breeding.