Genetic Variability for Haploid Production in Crosses Between Tetraploid and Hexaploid Wheats with Maize

Crosses were made between 14 wheat genotypes (11 tetraploid, 3 hexaploid) and a single Fl hybrid of maize that was used as the male parent. The experimental design consisted of randomized blocks with three replications. Plants were grown under controlled greenhouse conditions (day length 16 h and temperature 25 °C/15 °C, day/night). To enhance embryo survival, 2, 4-D treatment (10 mg/1) was applied to spikes 24 h after pollination with maize. Embryos were recovered from all tetraploid and hexaploid wheats at a rate of 2.09 to 26.76 per 100 pollinated florets. Haploid and doubled haploid plants were obtained from all hexaploid genotypes (T. aestivum) and from 5 of 11 tetraploid genotypes (T. turgidum var.). The most important point of these experiments was the ability to produce haploid plants from tetraploid wheat for two reasons: firstly, anther culture cannot be applied in tetraploid wheat (T. turgidum var.) due to the inefficiency of embryo formation and the high proportion of albino plants. Secondly, to date, crosses between tetraploid wheat and maize have resulted in embryo formation, but not in haploid plants.     

Factors Affecting Fertilization Frequency in Crosses of Triticum aestivum cv. 'Highbury'×Zea mays cv. 'Seneca 60';

Crosses of ‘Highbury’בSeneca 60’ were studied to determine the effect of four crossing procedures on the frequency of fertilization. The procedures were: 1) glumes of emasculated florets left intact, 2) glumes cut to expose the stigmas, 3) intact glumes given a 75 mg 1^-1 GA spray 2 h after pollination, and 4) cut glumes given a 75 mg 1^-1 GA spray 2 h after pollination. Within each treatment florets with developmental ages ranging from 3 days before an-thesis to 3 days after anthesis were pollinated. Fertilized florets were found in all four treatments. Overall, 20.2 % had only an embryo, 2.5 % had only an endosperm and 7.9 % had an embryo and an endosperm. Significant variation in the frequency of embryo formation, and hence in the frequency of potential plants, was found both between treatments and between developmental stages within treatments. The highest frequency (55.8 % of pollinated florets) was found in florets with intact glumes which had not received a GA spray and which had been pollinated on the day of anthesis. Cutting the glumes to expose the stigmas, pollinating earlier or later than the day of anthesis or application of a GA spray all tended to reduce the number of embryos. This was also the case when the total fertilization frequencies (embryos plus endosperms) were analyzed. The potential of wheat × maize crosses for wheat haploid production is discussed.     

Cytological Evidence for Fertilization in Hexaploid Wheat × Sorghum Crosses

Crosses were made between the hexaploid wheat ‘Chinese Spring’ (2n = 42) and the diploid grain sorghum ‘S9B’ (2n = 20). Sixty-nine out of 100 florets fixed 48 h after pollination contained an embryo, an endosperm, or both, a remarkably high frequency in view of the taxonomic distance spanned by the cross. Percentages of single or double fertilization ranged from 50 % to 91 % for individual spikes. The hybrid origin of the embryos was confirmed by examining zygotes from spikes fixed 25 to 27 h after pollination. Seven of the 8 zygotes in which chromosomes were sufficiently contracted to be counted contained 21 large wheat chromosomes and 10 much smaller sorghum chromosomes. The eighth contained 21 chromosomes from wheat and 20 from sorghum. Sorghum chromosomes did not appear to be attached to the spindle in zygote nietaphases and showed no evidence of movement towards the spindle poles in the single zygote anaphase found. Embryos with two or more cells invariably contained one or more micronuclei and metaphases in embryos with three or more cells contained only 21 wheat chromosomes showing that sorghum chromosomes were rapidly eliminated. Endosperm, when present, was always highly abnormal.     

Improved Techniques for Haploid Production in Wheat using Chromosome Elimination

Wheat × maize and wheat × pearl millet crosses have proved efficient for haploid production using various genotypes of wheat; 22 and 27 % of florets produced embryos. In favourable conditions 6—9 haploid plants per spike were produced. The following simplifications or improvements in technique are recommended: 1. Only a single treatment with an aqueous solution of dicamba or 2,4-D (50–100 ppm) for embryo stimulation in vivo; 2. Application by spraying or dipping the spikes; 3. Application time two to four days after pollination; 4. Embryo rescue 15 to 18 days after pollination; 5. Crosses without emasculation are possible if pollination occurs 1–2 days before anthesis. More than 450 haploids and some doubled haploid (DH) lines (after colchicine treatment in vitro) were produced using these methods. No hybrid plants, chromosome additions or substitutions were found.     

Inheritance of Cold Hardiness in Triticum aestivum × Synthetic Hexaploid Wheat Crosses

Synthetic hexaploid wheat, produced by combining tetraploid wheat (AB genome) with Triticum tauschii (D genome), was crossed to modern hexaploid wheat (Triticum aestivum ABD genome) in an attempt to introduce new cold hardiness genes into the common hexaploid wheat gene pool. The cold hardiness levels of F) hybrids ranged from similar to parental means to equal to the hardy parent, indicating that cold hardiness was controlled by both additive and dominant genes. As expected when dominant gene action is involved, differences between F₂ and parental means were smaller than comparable differences in the F., Frequency distributions of F₂—derived F₃ lines also suggested that dominant genes were involved in the control of cold hardiness in some crosses. Heritability estimates for cold hardiness ranged from 63 to 70 % indicating that selection for cold hardiness should be effective in populations arising from crosses between common and synthetic hexaploid wheat. However, high selection pressure on the progeny of crosses that included the most hardy T. aestivum, T. durum, and T. tauschii accessions as parents did not identify transgressive segregates for improved cold hardiness. These observations indicate that the close wheat relatives, sharing common genomes with T. aestivum, are not promising sources of new genes to increase the maximum cold hardiness potential of common hexaploid wheat.     

Addition of Colchicine to Wheat Anther Culture Media to Increase Doubled Haploid Plant Production

Chromosome doubling is critical for obtaining doubled-haploid plants from wheat (Triticum aestivum L.) anther culture. The most common doubling method applies colchicine to the plant. However, colchicine is phytotoxic and can induce a high frequency of plant death. In this experiment, anthers from two wheat genotypes (“Pavon 76” and ‘Centurk’) were placed on nine embryoid initiation media having three sugar sources (maltose, sucrose, and maltose + glucose) with three colchicine concentrations (0.0, 0.1, and 0.2 g · l^-1). Wheat starch was used as a gelling agent. After three days, the anthers were washed and moved to fresh media without colchicine. Increasing the colchicine concentration decreased the number of embryoids produced from 77.4 embryoids/100 anthers to 29.9 embryoids/100 anthers, but did not significantly affect the frequency of plant regeneration (0.49 green plants/embryoid to 0.40 green plants/embryoid), and increased the frequency of doubled-haploid plants (19.0 doubled-haploid plants/100 green plants to 72.3 doubled-haploid plants/100 green plants). Considering the total number of doubled-haploid plants produced, low levels of colchicine added to the initiation media were very effective.     

Androgenesis in Hexaploid Spring Wheat F2 Populations and Their Parents Using a Multiple-Step Regeneration System

The main goals of the wheat androgenesis experiment were to study the main phenomena of in vitro androgenesis in anther culture of F₂ populations (10) and their parents (6), to compare the usage of P-4 and C-17 media for the formation of embryo/callus and to demonstrate a new plant regeneration system. The P-4 induction medium was found to be significantly better than the C-17 in the number of responsive anthers (RA) and calli induced (CI) at the 1 % and 0.1 % level, respectively. Genotypic effect was evident in both RA and CI. The yields of F₂ populations in RA and CI were significantly higher than those of their parents regarding both media. The data confirmed the existence of heterosis for RA and CI in F₂ populations. The ratio of green/albino plant regeneration was more favourable in the C-17 derived embryo/calli than in the P-4 derived ones. The frequency of plantlet regeneration was enhanced in the group of unresponsive calli by the application of the multiple-step regeneration system. In this system the calli lacking well developed morphogenic structure were transferred to a new regeneration medium, containing a higher concentration of the same cytokinin, other cytokinin or basic medium, before the occurrence of irreversible changes in their physiology.     

Variation in Doubled Haploid Plants of Wheat Obtained through Wheat (Triticum aestivum) × Maize (Zea mays) Crosses

Variation was investigated in 110 doubled haploid (DH) lines of wheat derived from wheat × maize crosses. Field observation revealed visible variations in 15 DH2 lines such as extreme dwarfism, low seed fertility, alteration of spike type and stripes. Six agronomic traits, i.e., heading date, spike number/ plant, culm length, spike length, seed fertility and grain weight were statistically analyzed in the DH2 and DH3 generations. Out of the 88 DH2 lines/DH3 groups, 26 %/64 % showed significant differences from the parental variety in the means of one or more traits. Ranges of the DH3 lines were larger than those of the DH2 lines, except for spike number/plant. Furthermore, analyses of variance within and between DH lines showed the presence of heterogeneity/heterozygosity in the DH2 lines/plants. These results indicated the occurrence of gametoclonal variation in the DH lines. It is considered that most of the variations detected were due to the colchicine treatment rather than to the 2,4-D treatment or in vitro culture.     

不同小麦组合与玉米杂交产生单倍体的差异

实验采用３种类型、１５个小麦Ｆ１组合与玉米杂交种中单１２０进行杂交，研究不同小麦基因型单倍体产生的差异。     

玉米和小麦在光合诱导期间非光化学猝灭（qN）差异

为了比较玉米(Zea mays L.)（C4）与小麦 (Triticum aestivum L.)（C3）黑暗向光照变化过程中的光诱导适应性反应，对非光化学猝灭（Non-photochemical quenching, qN）的动态变化特征进行了观察和分析。结果表明，在15 min至24 h的不同暗适应处理中，根据qN的光适应的动态变化可分为F型（快速稳定型）、M型（中速稳定型）和S型（慢速稳定型）3种。在暗适应后的光诱导期间，小麦q_N主要表现出M和F型，玉米则表现出S和M型。通过对主要的荧光参数分析，小麦与玉米的F_v/F_m,qp和Φ_psII差异较小，而且qN稳定值有一定的差异，但TqNmex(q_N达到最大值的时间）和Tq_N（q_N达到稳定值的时间）玉米明显高于小麦，差异极明显。进一步分析经黑暗处理后光诱导的光合速率和气孔导度的动态变化，两作物虽然在增加的速率和稳定后的绝对值有差异，但达到稳定的时间无明显的不同。通过塑料膜包封叶片的方法阴止CO₂同化，而qN变化特征也基本不改变。这意味着短时间的C代谢不明显影响q_N对光诱导的反应。对光系统I反应中心P₇₀₃的典型氧化还原方式的测定和电子库容能力的分析也表明它们不是造成qN动态变化的主要原因。然而，对qNmex成分的分析证明高能态猝灭（high-energy state quenching,q_E）、转化猝灭（transition quenching,q_T)和光抑制猝灭（photoinhibitory quenching,q1)分别为55.6%、18.5%和25.9%，说明qE是导致玉米qN在暗处理后的光诱导表现出TqN_mex高、Tq_N长的主要原因。这意味着玉米（C4）在暗一光的变化中维持较高的qE，保持较高的类囊体质子梯度，有益于启动NADPH和ATP的形成。     

AFLP analysis indicates no introgression of maize DNA in wheat x maize crosses

Amplified fragment length polymorphisms (AFLPs) were used to follow the possible introgression of maize DNA into haploids of wheat as a side‐effect of exploiting wheat x maize hybridization for haploid production. AFLPs were generated with 64 MseI/ EcoRI and 64 MseI/ PstI primer combinations, and the AFLP profiles of haploids were tested against those of maize and of the regular wheat varieties involved in the crosses. On average, 45.1 and 110.7 fragments were produced per assay with the MseI/EcoRI and MseI/PstI combinations, respectively. Different numbers of fragments were produced for wheat and maize: an average of 81 in the haploid, 80 in the wheat parent, and only 67.1 in maize. No evidence was found for introgression of maize into the wheat genome. Three unique AFLP fragments were detected in haploids, which were not present in the parental wheat genotypes. These ‘novel’ AFLP bands in the haploids could be caused by nucleo‐cytoplasmic interaction in the hybrid zygote. Such instability in the wheat genome is defined as temporal, as it was not detected in further generations when colchicine‐doubled progeny of the haploids was tested for the presence of polymorphic fragments.     

Stability Analysis of Yield in Maize,Wheat and Sorghum and its Implications in Breeding Programs*

Six stability statistics: (b_i, s²_di, , , and ) were estimated for maize, wheat and sorghum in different environments by using three statistical models. The significant linear portion of genotype × environment interaction for maize indicates different hybrids responded differently to environments, whereas the non-significant genotype × environment interaction (linear) were found for wheat and sorghum suggest that all genotypes responded similarly as the environments change. However, the highly significant pooled deviations (deviation from regression) for all three crops make yield predictions from the model less reliable. When regression coefficients (b_i) were non-significant, s²_di, became an important statistic in estimating stability. It appears that the regression coefficient, b_i, was best used to estimate genotypic adaptability, whereas s²_di, for stability. Maize and sorghum had negative correlations between the mean yield and stability statistics, s²_di, , and , suggesting that high yield and stability are not mutually exclusive in the range of environments used in this study; however, such correlations did not occur in wheat. Thus, maize and sorghum hybrids with high yield potential and high stability could be identified and selected. Correlations between mean yield and b_i, or , were positive and significant for maize and sorghum but were non-significant for wheat, indicating that such relationships may be species specific. Under a given set of testing environments, the stability ranking associated with each maize hybrid is correlated to and depends on other hybrids included in the analysis.     

Chromosomal Location of Dimeric Phosphatase Structural Genes in Hexaploid Wheat

Structural genes for leaf dimeric phosphatases (E.C. 3.1,3.2.) have been located on wheat chromosome arms 7BL and 7DL. No gene was found on 7AL. The results obtained pointed to the existence of two kinds of wheat phosphatases (monomers and dimers j located on different homoeologous groups, which is in agreement with results found in Secale cereale and Agropyron intermedium. These results allow homoeologous relationships to be established through the classification of dimeric phosphatases and also provides another useful genetic marker for the 7BL and 7DL arms., They also provide further support for die concept of the conservation of gene synteny groups within the Triticnae.     

The Growth in Culture of Detached Ovaries of Wheat (Triticum aestivum L. em. Thell)

Viable embryos have been recovered from fertilized wheat (Triticum aestivum) ovaries detached on the day of, or up to 7 days post-, anthesis and cultured aseptically fur up to 18 days. The most significant factors in determining the yield of embryos was the plating density, age and complexity of the explants. 26% of ovaries excised on the day of anthesis produced viable embryos if cultured as pairs of florets. The potential use of detached ovary cultures in gametophyte microinjection experiments, rescuing embryos from wide crosses and in chemically manipulating the early stages of embryo development are discussed.     

Detection of Resistance to the Russian Wheat Aphid in Hexaploid Wheat

The Russian wheat aphid (RWA), Diuraphis noxia (Mordvilko), poses a serious threat to wheat (Triticum aestivum L.) production in many parts of the world. This research was initiated to evaluate wheat accessions for detection of resistance to the RWA. Over 12,000 wheat cultivars and plant introductions (PIs) from the USDA-ARS National Small Grains Collection were evaluated for reaction to RWA feeding damage. Twenty-nine PIs from Iran, Afghanistan, and the former Soviet Union, of various agronomic backgrounds were identified as having moderate to high levels of RWA resistance. This information is useful to wheat breeders searching for sources of resistance to the RWA to incorporate into their breeding programmes.     

Field Performance of Androgenetic Doubled Haploid Spring Wheat Lines in Comparison with Lines Selected by the Pedigree System

A total of 316 doubled haploid lines (DH) of spring wheat were compared with 621 lines selected in a pedigree system (PS) under field conditions in a breeding nursery. The lines originated from 21 crosses and the samples tested represent mean values for variables comprising the main breeding goals such as disease resistance, baking quality, and agronomic traits. In general, the DH lines were later in days to heading and shorter than PS lines of different homozygosity levels. Except for leaf rust (Puccinia recondite), the DHs were more resistant to the artificially infected diseases such as powdery mildew (Erysiphe graminis), stripe rust (Puccinia striiformis), and septoria nodorum blotch (Septoria nodorum). These differences, which were relatively small for practical selection purposes, were probably due to easier and more precise disease assessment of the homozygous DH lines. For quality characters, the DHs had a higher protein level, while the rest of the parameters were similar for both origins. The data analyzed suggest so far that androgentic doubled haploids in spring wheat are very similar to lines selected in a pedigree system in respect to all the agronomic characters tested. However, the DH lines were produced in a much shorter period of time. It is suggested that androgenetic doubled haploids be produced from F₁ hybrids and that the well-established bulk method should continue to allow selection bulk method should continue to allow selection for rare recombinants as soon as homozygosity is reached.     

Agronomic Performance of Hexaploid Wheat Lines Derived from Direct Crosses between Wheat and Aegilops squarrosa1)

Direct introgression of genes from diploid Aegilops squarrosa L into hexaploid wheat (Tnticum aestivumL.) is an efficient way of developing stable, hexaploid lines with unique, useful genes. 39 such lines derived front direct crosses to determine the effects of Ac. squarrosa germplasm were field-tested on productivity. Whereas some lines were agronomically inferior to the recurrent parent, others periormed at an equall or better level. In addition to the larget traits (Hessian-fly and leaf-rust resistance), other traits, including soilborne-mosaic-virus resistance and protein concentration, were improved.     

A simple technique for the identification of embryo-carrying seeds from wheat × maize crosses prior to dissection

Embryo rescue is an important step in the wheat x maize system of induced haploidy in wheat. On average, only one-third of the seeds carry embryos, but they all have to be dissected because no morphological distinction is available to distinguish between seeds with and without embryos. We here report a simple technique in which immature seeds from wheat × maize crosses are placed on a transparent surface and illuminated from above. The free-floating embryo, settled at the bottom of the seed, can be seen from below or in an image reflected on a plane mirror. Using this technique 97.8% of the embryo-containing seeds could be detected prior to dissection. The technique will be useful in reducing the time and labour involved in embryo culture, thereby scaling up haploid production.     

Effects of Wheat Chromosome 1D Telosomes on Hybrid Seed Development in Wheat × Rye Crosses

Pollination of ‘Chinese Spring,’ monosome 1D plants with rye results in failure of hybrid seed development in a proportion of the F₁ seeds corresponding to the transmission rate of the nullisomic 1D egg cells. Development and viability of these hybrid seeds closely resemble that normally observed in T. aurum× rye crosses. Using ‘Chinese Spring’ chromosome ID telosomic plants in crosses with rye, it was possible to illustrate that the observed effect was associated with the long arm of this chromosome.     

The Response of Anther Culture in a Genetically Wide Material of Winter Wheat (Triticum aestivum L.)

In a screening experiment with 215 different winter cultivars of Triticum aestivum L. for response in anther culture, pollen embryos or callus were obtained from 200 different cultivars and green plants from 93 different cultivars. On average, from the whole material, 1.3 green plants were obtained per 100 anthers cultured. Variance components estimated from replicates with selected parts of the plant material indicated that for the formation of pollen embryos from anthers, interactions between genotypes and replications were dominating, accounting for 45 to 50 per cent of the variation. Main effects from genotypes were less prominent, accounting for 25 to 30 per cent of the variation, and the replications showed relatively little effect, accounting, for about 4 to 12 per cent of total variation. Regeneration of plants from pollen embryos or callus was not significantly influenced by either genotype or replicates. The frequencies of plants regenerated being green, however, were influenced from both genotype and environments contributing 42.5 and 34.9 per cent of the total variation respectively, while only minor interactions between genotype and environments were indicated for this character.