Production of intraspecific aneuploids in the genus Solanum. III. Intraspecific aneuploids

Summary The triploids recovered from 4x×2x crosses in three Solanum species were very vigorous and although few seeds/fruit were obtained when the triploids were crossed to diploids, the extensive crossing programme produced sufficient seed of four species cph, chc, pnt and tar. The average seed set for the 3x-2x crosses was an extremely low 3.5 seeds/fruit.Approximately 90% of the progeny of the 3x–2x crosses were aneuploids with chromosome numbers of 2n=25–29. The frequency of the aneuploids in the three species that were studied was chc 93%, pnt 94% and tar 92%. The aneuploids of chc and tar were extremely vigorous and fertile and they were used as females in crosses to the marker stocks. The aneuploids of pnt were vigorous, but the crossability barrier of pnt prevented their use in crosses to the marker stocks. A number of the aneuploids produced seed upon being selfed, but the ability to produce self seed may be related to the pseudo-compatibility of the parental clones. In only 7 aneuploids was there an indication that the self fertility was due to overcoming the self-incompatibility barrier as a result of competition-interaction of the aneuploid gamete.Plants in the families from the triploid-diploid crosses had a tremendous amount of variation in morphological characteristics (leaf shape, size and color; berry shape, color and degree or verrucose spotting: and plant habit and vigor). A large portion of variation exhibited in these families was due to the normal genetic segregation of the heterozygous parents. It was impossible to distinguish the aneuploids from their diploid sibs especially those having only one or two additional chromosomes because they were as vigorous and fertile as the diploids.There were some preliminary indications of the existence of distinct morphological characteristics among the aneuploids (separate petals, long berries, and extreme verrucose berries). However, there was no indication that these traits were due to the dosage effect of the extra chromosome. If it should be determined that this was true, it would be extremely useful in associating genes with chromosomes and determining the phenotypic effects due to the presence of an additional chromosome.Cooperative investigations of the College of Agriculture and Life Sciences, University of Winconsin and the Agricultural Research Service, U.S. Department of Agriculture, Madison, Wisconsin. Supported in part by grants from the Research Committee of the Graduate School, University of Wisconsin.First and third author respectively, formally Graduate Research Assistant Department of Horticulture, University of Wisconsin, Madison, and Geneticist Agriculture Research Service, U.S. Department of Agriculture, University of Wisconsin, Madison. Present address: Department of Genetics. University of Wisconsin, Madison, Wisconsin 53706 and Head Breeding and Genetics Department, International Potato Center, Lima, Peru.     

Aneuploids of perennial ryegrass (Lolium perenne)

Summary Aneuploid plants of perennial ryegrass (Lolium perenne L.) with 2n=15 to 30 chromosomes were obtained by crossing a near-triploid (2n=3x+1=22) with a diploid or on open-pollination with diploids and tetraploids. Aneuploids occurred with a frequency of 83% in near triploid × diploid progeny and 92% on open-pollination with diploid and tetraploid plants. Aneuploid plants with 15 to 18 chromosomes resembled diploids in morphology and those with 19 to 30 chromosomes were akin to tetraploids. Meiotic studies suggested that most aneuploid plants resulted from transmission of aneuploid egg cells (n=8 to 23). Aneuploid plants with 2n=27 to 30 chromosomes in the progeny of 22×14 cross originated from unreduced egg cells. Plants with 19 to 21 chromosomes were recovered only by immature seed culture. Aneuploid plants with 26 to 30 chromosomes and triploids (2n=21) had higher pollen fertility and bigger seeds than plants with 15 to 22 chromosomes.     

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     

Bilateral polyploidization inDactylis glomerata L. subsp.lusitanica: occurrence,morphological and genetic characteristics of first polyploids

Summary InDactylis glomerata L. subsp.lusitanica, triploid and tetraploid plants were obtained by bilateral sexual polyploidization in crosses between diploid parents known to produce 2n gametes. The polyploid and diploid progeny were compared for allozyme diversity (allele number and heterozygosity), phenological (pollen fertility, inflorescence emergence date), cellular (stomatic cell size) and morphological characters (vegetative biomass, seed weight, total seed number per plant, panicle number and seed number per panicle length) in four contrasting growing conditions. Stomatal cell size and the total number of alleles were significantly lower in diploids than in polyploids which sometimes recovered more than two distinct alleles at a locus via 2n gametes possessing heterozygous loci. Pollen fertility was similar in diploids and tetraploids, but significantly lower in the triploids. Seed weight was significantly higher in polyploids, although the average number of seeds per panicle length and the total number of seeds was higher in the diploid sibs. No phenological differences occurred among ploidy levels whereas such differences were observed inDactylis in all the natural areas of sympatry between diploids and their related polyploids. Our results provide evidence for an instantaneous effect of polyploidization on certain traits and suggest that natural selection would act subsequently on several other traits to allow the establishment of new polyploids.     

Identification of tetraploid regenerants from cotyledons of diploid watermelon cultured in vitro

Summary Adventitious shoots were obtained from the diploid watermelon [Citrullus lanatus (Thunb.) Matsum. & Nakai] cultivars Dixielee, Jubilee II, Mickylee, Minilee, and Royal Sweet by culturing excised cotyledons on shoot regeneration medium for six weeks. Tetraploid and diploid regenerants were identified by counting the number of chloroplasts per guard cell pair from leaves of regenerated plants. Cross fertilization of putative tetraploids with diploid pollinators and the production of triploid seed confirmed the efficacy of this approach. The mean number of chloroplasts for tetraploid regenerants was 19.1 whereas diploids averaged 11.2. These values were similar to tetraploid and diploid plants from seed. Ovary diameter, petal, and anther diameter of male flowers, and leaf length by width ratio were also good indicators of plant ploidy. Progeny obtained from self-fertile tetraploids of Mickylee were crossed with various diploid pollinators to produce triploid hybrid seed. All triploid plants from tissue culture-derived tetraploids produced fruit comparable in quality to fruit produced by currently-available triploid hybrids, demonstrating that in vitro tetraploid induction can be used to produce high quality tetraploid plants for use in triploid hybrid seed production.     

Crossability relationships of andean potato varieties of three ploidy levels

Summary The formation of triploids in 4x×2x crosses was studied in primitive potato varieties from the Andes of Peru and Bolivia. Triploid frequencies were as high as 80–100% in some crosses, although the actual number of seeds was low. The triploid block varied according to the tetraploid parent, and was never entirely absent. The likelihood of triploid formation was studied in 4x×2x crosses where pollen from both tetraploids and diploids of known genotype was used. Even though the diploids were chosen for their non-production of 2n gametes, as detected in pollen grain size, 66% of the hybrids were tetraploid. The amount and probability of gene-flow between the tetraploid and diploid gene-pools, either directly through 2n gametes or through triploid bridges was examined. Most of the 20 cultivars of S. x chaucha produced seed in crosses with tetraploids and diploids, although the fertility of the latter crosses was four times higher. The female fertility of some of the triploids thus allows them to exchange genes with the tetraploids and diploids, although gene-flow occurs more rapidly through direct transfer from diploids to tetraploids via 2n gametes.     

Isozyme variation between diploid and tetraploid cytotypes of Glycine tabacina (Labill.) Benth.

Summary Glycine tabacina (Labill.) Benth. is a wild perennial species related to the cultivated soybean, G. max (L.) Merr. It is composed of diploid (2n=40) and tetraploid (2n=80) cytotypes. Currently, to differentiate the cytotypes, plants are grown out in the greenhouse and chromosome counts made on pollen mother cells. It is a laborious and time consuming process. The objective of this study was to determine whether electrophoretic techniques could be utilized to separate the cytotypes. Electrophoretic examination of seven isozyme systems from seed of 67 G. tabacina accessions revealed banding patterns that could be used to differentiate between diploid and tetraploid cytotypes in the species. Among the tetraploid accessions, the number of bands observed were always greater than the diploids. Some tetraploid banding patterns consisted of bands similar to the diploid tabacina and/or additional bands previously identified in other Glycine species. The patterns of isozyme multiplicity and variation in the tetraploid tabacinas suggests more than one mode of origin for the tetraploids.     

Note on the probable cause of the occurrence of tetraploid plants in commercial triploid varieties of sugar beet

In crossing diploid male-sterile beets (18 chromosomes) with a tetraploid pollinator (36 chromosomes) it was established that of 532 individuals from this cross 8.4% were diploid, 83.9% triploid and 7.7% tetraploid.The most reasonable explanation was that the diploid male-sterile plants had given a number of unreduced egg cells. This could be put to a test by crossing diploid male-sterile plants with normal diploids. The average number of triploids found by chromosome counts was 2.4%; in a normal diploid no triploids were found.The type of cytoplasm associated with the male-sterility may lead to abnormal egg cells.Trirave (Triplex) with 84% of triploids, represents an enormous advance over normal polyploid types, which very commonly have no more than 40 per cent triploids and very rarely have more than 50 per cent.Bush Johnsons Limited and D.J. van der Have n.v.Paper read on the XXIInd Winter Meeting of the Institut International de Recherches Betteravières on February 24th, 1959.     

大花香石竹多倍体育种研究

对102个大花香石竹品种的染色体倍性进行鉴定,二倍体、三倍体和四倍体品种分别占77%、6%和17%,当前生产上的主栽品种马斯特、达拉斯和卡曼（具有大花苞性状）都是二倍体品种,71%的四倍体品种为花边复色类型。初步认为部分香石竹四倍体栽培品种是远缘杂交的后代。通过用秋水仙碱处理香石竹品种马斯特的试管苗,获得两株四倍体植株,其花蕾直径增大,而节间变短。     

The importance of two-seeded pods in red clover (Trifolium pratense L.)

Summary The effect of selection for two-seeded pods in diploid and tetraploid populations and the effect of chromosome doubling on two-seededness were studied. The material obtained was examined to find out how important two-seeded pods are for seed set, grain weight and seed yield.In a mixoploid plant the percentage two-seeded pods in the tetraploid heads was far smaller than that in the diploid heads. Only when the character had developed relatively strongly in the diploid, did it exceed a very low value.By selection of individual plants in the best families of a diploid group of plants the percentage two-seeded pods could be raised from on average 7.4 to 30.6% as early as the following generation. Similar selection in tetraploid material with 2.5% two-seeded pods did not increase the percentage.The reduction of grain weight of seed in two-seeded pods compared with that in one-seeded pods was greater in the tetraploid plants than in the diploids.No effect of two-seededness on seed yield was found in our material. A marked effect on seed-set was observed in a diploid population (286 plants) when the percentages two-seeded pods rose about 16%.As the correlation between the calculated number of heads and the seed yield per plant is high, it may be adopted that in our material the number of heads per plant was decisive of seed yield. This will contribute to the small effect of two-seededness on the seed yield of the plant.The results of the present study showed that the production of tetraploids with a high percentage two-seeded pods will take more advantage of selection of diploid material followed by doubling of chromosomes than of direct selection in the tetraploids.Protrusion of pods in heads indicates a very high percentage two-seeded pods. This will enable relatively rapid selection for two-seeded pods.A quite small percentage two-seeded pods also occurred in two tetraploid populations that had not been selected for two-seeded pods.     

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.     

Unreduced pollen frequencies versus hybrid production in diploid-tetraploid Vaccinium crosses

Summary Reciprocal crosses were made between highbush Vaccinium corymbosum L. cultivars (4X) and high, medium and low frequency diplandroid pollen producing diploid clones of V. elliotti (Chapm.) and diploid pubescent forms of V. corymbosum. Pollination of 3,342 flowers produced 15 hybrids-11 tetraploids and 4 triploids. All V. elliottii hybrids were tetraploid while each successful diploid-tetraploid V. corymbosum cross produced at least one triploid. The ratio of tetraploid hybrids to 100 pollinations (H: 100P) was considered to be the best criterion for assessing crossability. Vaccinium elliottii produced a total of 8 4X hybrids while 2X V. corymbosum only produced 3. Vaccinium elliottii was more successful as a female parent and diploid V. corymbosum only produced hybrids as male parents. High frequency diplandroid clones showed a lower mean crossability as pollen parents (H: 100P=0.2) than medium frequency diplandroid clones (H: 100P=0.8). As predicted, 0% diplandroid clones wer unsuccessful as pollen parents. Unreduced pollen production was essential for tetraploid hybrid production in 4X×2X crosses. There appeared to be no consistent, inherent pattern between 2n pollen and 2n egg production in a clone nor was a tetraploid genotype effect on crossability consistently observed.Paper No. 10781 of the Journal Series of the North Carolina Agricultural Research Service, Raleigh, NC., 27695-7601     

Influence of flower and flowering characteristics on seed yield in diploid and tetraploid red clover

Tetraploid red clover (Trifolium pratense L.) often produces insufficient seed to be economically interesting. Numerous studies have identified poor pollination due to long corolla tubes as its cause, but other studies disagree. Therefore, we investigated seed yield, corolla tube length and flowering traits in 244 plants from five diploid and five tetraploid cultivars during 2 years. Tetraploids produced fewer seeds, fewer flower heads and fewer seeds per head, but an equal number of flowers per head when compared to diploids. Although corolla tubes were longer in tetraploid than in diploid red clover (P < 0.001), no correlation between corolla tube length and seed yield was detected. Therefore, the corolla tube length is likely neither the main nor only cause of low seed yield in tetraploid red clover. Instead, three putative causes for low seed yield were identified: tetraploids produced less inflorescences per plant, possessed a lower degree of determinacy and had markedly less seeds per head when compared to diploids. Possible causes for such differences include lower pollen viability and higher rates of embryo abortion.     

不结球白菜同源四倍体苏州青的诱导鉴定

为获得高产优质的不结球白菜四倍体苏州青,本研究使用浓度分别为0.1%,0.2%及0.3%的秋水仙素溶液,点滴处理二倍体苏州青幼苗植株的子叶生长点,进行5次处理后,通过形态学和细胞学方法筛选鉴定四倍体苏州青植株,并比较二、四倍体苏州青的农艺性状及品质差异。结果表明：秋水仙素浓度为0.2%时加倍效果最好。形态解剖学鉴定结果表明：四倍体在株型、叶片、花、种荚、种子大小都有较明显的变大趋势;四倍体植株叶片气孔较二倍体变大,气孔密度变小;同时四倍体植株花粉粒比二倍体更膨大,更不规则;细胞学鉴定结果表示,四倍体植株染色体数为40条,二倍体植株为20条;通过流式细胞仪鉴定,四倍体植株荧光强度约为二倍体的2倍。通过品质比较分析,结果显示四倍体苏州青的各项生理指标含量较于二倍体均有不同。通过农艺性状比较,单株质量、叶宽、叶厚都有一定程度的增加。本研究利用秋水仙素操作技术,抑制有丝分裂后期纺锤丝的形成,实现染色体加倍,创制不结球白菜新种质,为选育四倍体不结球白菜提供理论基础和种质资源。     

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.     

Cytogeography and variation of stomatal size of Paspalum Glaucescens (Gramineae; Paniceae) in Southern Brazil

Paspalum glaucescens belongs to the informal group Plicatula, reproductively characterized by the dominance of tetraploid apomitic lines in most of its populations, with rare diploid, sexual counterparts. The species shows high phenotypic variation. Twenty nine Southern Brazilian accessions were cytologically and morphologically analysed. Most of the accessions were tetraploid (2n = 4x = 40). Meiotic study of three tetraploids showed their irregular behaviour. Eight accessions presented the diploid level (2n = 2x = 20).This can be considered a very high frequency of diploids in a member of the Plicatula group. While the tetraploids are concentrated at the highest elevations, diploids were detected mostly in the lowlands. Average stomatal sizes were quite distinct on different ploidy levels, being larger in tetraploids. This revised version was published online in July 2006 with corrections to the Cover Date.     

Artificially induced polyploidization in Humulus lupulus L. and its effect on morphological and chemical traits

Chemically induced polyploids were obtained by the colchicine treatment of shoot tips of Humulus lupulus L. ‘Sybilla’. Flow cytometry revealed that most of the treatments resulted in the production of tetraploids. The highest number of tetraploids was obtained when explants were immersed in 0.05% colchicine for 48 h. A field experiment was conducted to compare diploid and tetraploid plants and assess the effect of genome polyploidization on the morphological and chemical characteristics. Tetraploids showed significant differences in relation to diploids. They had thinner and shorter shoots. The influence of chromosome doubling was also reflected in the length, width and area of leaves. The length of female flowers in the tetraploids was significantly shorter than that observed in diploids. Tetraploids produced a diverse number of lupuline glands that were almost twice as large as those observed in diploids. The most distinct effect of genome polyploidization was a significant increase in the weight of cones and spindles. Contents of major chemical constituents of hop cones was little affected by ploidy level. Total essential oils were significantly lower than those in diploids. However there was a significant increase in the proportion of humulene, caryophyllene and farnesene, oils desired by the brewing industry.     

Evaluation of FDR diploid and tetraploid parents in potato under two different day-length environments

Five tetraploid clones of potato were crossed to each of eight diploid first division restitution (FDR) 2n pollen producers and four tetraploid males using a line × tester mating design. A total of 59 families were obtained and evaluated under long days at Rhinelander (USA). A subset of 49 families were grown under short days at four Peruvian locations. Tuber yield (kg per plant) of 4x-2x families was similar to, or greater than, that of 4x-4x families in short day environments. The diploid clones had breeding values greater than or equal to those of the tetraploid clones. Only families derived from FDR 2n pollinators combined high tuber yield with stability and high dry matter content. Reciprocal recurrent selection would be the best breeding scheme, in which the diploids are testers of the tetraploids and vice-versa. Intra-population improvement could be achieved through phenotypic recurrent selection. The best materials from both breeding pools could then be crossed to produce tetraploid hybrids through unilateral sexual polyploidization (4x-2x crosses).     

The breeding of the cultivated potato species solanum x juzepczukii and S. x curtilobum II. The resynthesis of S. x juzepczukii and S. x curtilobum

Summary The possibility of combining anew the genomes of wild and cultivated progenitors of triploid S. x juzepczukii and pentaploid S. x curtilobum by following the known evolutionary pathway of these species was investigated.The resynthesis of S. x juzepczukii was easy, and a wide range of synthetic forms was bred. Among these were forms with higher frost resistance (-5°C) than has been found in natural S. x juzepczukii. The total tuber glycoalkaloid content of several synthetic hybrids was lower than or as low as that of natural clones. Most synthetic hybrids were more vigorous than natural S. x juzepczukii and produced about the same types of tubers as are found in the natural range of variation. The best diploid parents were found in the species S. goniocalyx.The attempt to resynthesize pentaploid S. x curtilobum has not been successful but tetraploid plants were obtained in the process. An explanation for the occurrence of tetraploids resulting from triploid x tetraploid and/or diploid crosses is offered.The newly bred tetraploids contain at least one genome from S. acaule (possibly two) and hybridize easily with ssp. andigena. They thus provide a means for the transfer of S. acaule germ plasm into the tetraploid cultivated gene pool which would profit from the frost resistance of S. acaule.     

Breeding of polyploid spinach

2x, 3x and 4x forms of spinach were compared in 1959, 1960 and 1961.At the same stage of development the tetraploids outyielded the diploids in total green matter and leaf blades in 1959 and 1961. In 1960 there was no significant difference. This year the tetraploids had a very small increase in green matter in the period between the two harvest dates. Yield of dry matter was determined in 1960 and 1961, and in both years the tetraploids outyielded the diploids. The rate of development decreased with increasing degree of ploidy. Compared at the same age, when the diploids were about to start bolting they were superior to the polyploids with regard to total yield, yield of leaf blades and yield of dry matter.The onset of flowering was considerably delayed in the polyploids. Generally the triploids were intermediate between the diploids and the tetraploids. The triploids and tetraploids had a larger proportion of monoecious plants, and a smaller proportion of males than the diploids.Report No. 27