Pollen sterility induced by ethrel and its utilization in hybridization of wheat

Summary In 1974 and 1975 gametocidal effects of Ethrel were investigated under field conditions in the spring wheat cv. Zlatka. In experimental plants grain setting per spikelet and ear as well as ability to be cross-pollinated were ascertained.As compared with the control, treated variants achieved more than 90% sterility. Female fertility was not affected and cross-pollination was possible. Genetical check showed that the best variants, when cross-pollinated, produced 50–55% hybrids; grain setting, however, was brought down by more than 37%. As a result of Ethrel treatment haulm length was reduced and the vegetation period was somewhat prolonged.     

Induction of male sterility in wheat using organic ligands with high specificity for binding copper

Summary Because copper is extremely important to the development of normal polllen, an attempt was made to induce male sterility in wheat by applying specific copper-binding ligands to wheat plants. Four different chelates were used at two rates in three methods of application. All four chelates, cupferron, neocuproine, benzotriazole and cuprizone, reduced grain yield at high concentration applied to the soil at sowing but benzotriazole was most effective, even when applied at late tillering to either soil or foliage, and it also reduced yield to a lesser extent when applied at low concentration. At high concentration of benzotriazole (50 mg kg^-1 of dry soil) the percentage of pollen staining with I₂/KI was very low (0–7%) depending on method of chelate application), and this soil treatment resulted in complete male sterility. The appearance of the pollen, anthers, grain, ears and leaves in many cases mimicked that of normal copper deficiency, and also that caused by other recognised gametocides. These results raise the question of whether binding of copper or some other disturbance of copper metabolism may be the mechanism by which andro-gametocidal chemicals work and if so, dictate a theoretical basis for selecting such chemicals for testing.     

A comparison of cytoplasmic and chemically-induced male sterility systems for hybrid seed production in wheat (Triticum aestivum L.)

A comparison of two male sterility systems was carried out in wheat for hybrid seed production and hybrid performance. Seventeen hybrid combinations based on Triticum timopheevi cytoplasm were compared with their genetically equivalent CHA-facilitated combinations. The same set of restorer lines was used as parents in CMS- and CHA-based hybrids to maintain genetic equivalence. In the first experiment aimed at study of female line behavior and crossed seed production, the CHA treated lines showed significantly shorter heights whereas CMS lines were similar to the control. The two systems were equally effective in sterilizing rate. The outcrossing percentage of the CMS lines was almost twice that of the CHA treated lines. Thousand-grain weight of the crossed seeds on CMS lines was greater than on the CHA treated lines and control. On average, the germination percentage of seeds on CMS lines was double that of the CHA treated lines and the percentage of effective outcrossed seeds in CMS lines was 3 times more than that from CHA treated lines. The second experiment was conducted to examine the yield performance of the hybrids derived from the two systems of male sterility. A total of 40 entries including 20 hybrids and 20 parents were evaluated in the experiment. The mean grain yield of the CMS-based hybrids was greater than that of the CHA-based hybrids,the B and R lines. All the CMS-based hybrids showed significantly higher grain yields than their better parents whereas all the CHA-based hybrids, except two,showed no significant yield increases over their better parents. Possible reasons for differences in CMS- and CHA-based hybrid performance are discussed. This revised version was published online in July 2006 with corrections to the Cover Date.     

Photoperiod-sensitive cytoplasmic male sterility in wheat with Aegilops crassa cytoplasm

Summary Triticum aestivum cv. Norin 26 with Aegilops crassa, Ae. juvenalis or Ae. vavilovii cytoplasm (all D² type) has been studied relative to its photoperiodic response of male sterility and fertility restoration patterns. Alloplasmic lines of Norin 26 with a D² type cytoplasm showed almost complete male sterility under long-day conditions (15 h), but high male fertility under short-day conditions (14.5 h). No significant influence of temperature on reduction in male fertility was observed. Thus, this type of male sterility is called photoperiod-sensitive cytoplasmic male sterility (PCMS). The PCMS is expressed in the form of pistillody of stamens. Histological studies revealed that there were incomplete ovule-like structures instead of tapetal cells and pollen grains in the pistillate stamens. The floret differentiation stage of the plant is the stage that is sensitive to photoperiod. The PCMS can be used as a new means for hybrid wheat production, named two-line system. The PCMS line is maintained and multiplied by self-fertilization under short-day conditions, and hybrid seed can be produced by crossing the PCMS line with a pollinator line under long-day conditions. In contrast to the system of hybrid wheat production using the T. timopheevi cytoplasm, the present system requires only PCMS and pollinator lines.Abbreviation CS Chinese Spring - N26 Norin 26 - PCMS photoperiod-sensitive cytoplasmic male sterility     

Induction of a mutation in the male fertility gene of the preferentially transmitted Aegilops sharonensis chromosome 4S1 and its application for hybrid wheat production

Summary Wheat plants nullisomic for chromosome 4B are male sterile due to the absence of the male fertility gene Ms1. However, plants in which chromosome 4B has been substituted by the preferentially transmitted chromosome 4S¹ of Ae. sharonensis are male fertile due to the compensating effect of Ms4 on the alien chromosome. This substitution line has been mutated and three recessive mutation of Ms4 have been selected. Plants homozygous for these mutations are male sterile. The implication of these mutations for hybrid wheat production is discussed.     

Mutations blocking sensitivity to gibberellic acid promote ethylene-induced male sterility in wheat

Summary Ethylene is known to perturb normal reproductive development in wheat, particularly the development of functional pollen. Two experiments were carried out to test the hypothesis that increasing insensitivity to gibberellic acid (GA), conferred by the Rht genes, would be associated with increased male sterility in ethrel or Cerone^®-treated wheat. Wild type (WT=rht1/rht1, rht2/rht2), single dwarf (SD=Rht1/Rht1, rht2/rht2 or SD=rht1/rht1, Rht2/Rht2), and double dwarf (DD=Rht1/Rht1, Rht2/Rht2) near-isogenic lines in six genetic backgrounds were treated with ethrel or Cerone^® at the late tetrad to early uninucleate stage of pollen grain development. Ethrel induced pollen abortion in all genotypes but was highest for DD (41% above background) followed by SD (20%), and then WT genotypes (10%). Spikelet fertility decreased as the number of Rht alleles increased in response to ethrel or Cerone^® treatments. Expressed as a percent of controls, spikelet fertility was 56% for WT, 42% for SD, and 29% for DD. The consistent linear relationship between the number of Rht alleles and sensitivity of ethylene-induced male sterility suggests that GA and its recognition may exert a stabilizing effect in pollen development in the presence of stress or an ethylene shock.Paper No. 764 of the Department of Plant Breeding and Biometry, Cornell Agricultural Experiment Station.     

Gametocidal properties of RH-531, RH-532, RH-2956, and RH-4667 on spring wheat (Triticum aestivum L.)

Summary Several chemicals were evaluated for their gametocidal properties to control pollination in hybrid wheat (Triticum aestivum L. em Thell.) seed production. In a preliminary study, four chemicals were applied to three cultivars of spring wheat at two application times. RH-532 was the most effective of the four gametocides, reducing fertility to a 0 10% level for all three cultivars.In a second experiment, RH-532 was applied to one cytoplasmic male-sterile and three normal spring wheats at four rates (0.56, 1.12, 2.24, and 4.48 kg/ha) and at two times of application. Alternate strips of pollinator were planted throughout the experiment. All treatments decreased fertility. plant height, and spike length of the three normal wheats. RH-532 did not inhibit spike emergence of two semidwarf cultivars or one of conventional height. Cultivars did not respond similarly to treatments.Yields of cross-pollinated seed were only 1 21% of the normal yield when fertility levels on treated plants reached 0 10%. Hybrid seed content of the harvested seed ranged from 2 55%. On the treated male-sterile line, yield was significantly reduced in comparison with the open-pollinated check. Inhibitory properties of this chemical suppressed the fertilization potential of the female as well as inducing male sterility.The differential response of genotypes to treatments indicates that cultivars should be screened for male and female sensitivity to this chemical. If differential sensitivity is found, breeding and selection to improve this trait may be possible. Different environmental conditions may alter the response of cultivars to this chemical, with less effect on the female structures.Published with the approval of the Director, North Dakota Agricultural Experiment Station, as journal article No 71a.     

Response of wheat tillers at different growing stages to gametocide treatment

Summary The effects of the gametocides Ethrel, RH-532, and RH-2956 on wheat tillers at various stages of development were studied. The gametocides were applied to Anza and Yecora 70, two spring wheat (Triticum aestivum L.) varieties, at three rates of foliar application when the main tillers were at meiosis. Ethrel was less effective in causing sterility than RH-532 and RH-2956. Anza showed greater induced sterility than Yecora 70 when treated with these gametocides. Ethrel affected only late tillers treated at or before meiosis. RH-532 and RH-2956 at the high application rate were equally effective for Anza. RH-2956 at the high application rate induced uniform and maximum sterility in early and late tillers of both Anza and Yecora 70.     

Development of photoperiod-sensitive cytoplasmic male sterile (PCMS) wheat lines showing high male sterility under long-day conditions and high seed fertility under short-day conditions

A “two-line system” using photoperiod-sensitive cytoplasmic male sterility (PCMS) caused by Aegilops crassa cytoplasm under long-day photoperiods (≧15 h) has been proposed as a means of producing hybrid varieties in common wheat (Triticum aestivum). The PCMS line is maintained by self-pollination under short-day conditions, and hybrid seeds can be produced through outcrossing of the PCMS line with a pollinator line under long-day conditions. Our previous studies revealed that PCMS lines showing complete male sterility under long-day conditions are necessary for practical hybrid wheat breeding, especially to obtain high hybrid purity in F₁ seeds. Furthermore, practical PCMS lines should have high seed fertility under short-day conditions, which is associated with female fertility. Wheat cv. Norin 26 with Ae. crassa cytoplasm exhibits high seed fertility under short-day conditions, and cv. Fujimikomugi with Ae. crassa cytoplasm shows high male sterility under long-day conditions. Here we developed practical PCMS lines derived from the F₁ generation of Norin 26 and Fujimikomugi (with Ae. crassa cytoplasm) that were then backcrossed to elite wheat lines.     

Temperature effects on seed germination and expression of seed dormancy in wheat

Because preharvest sprouting decreases quantity and quality of wheat grain, researchers need effective protocols to assess response to preharvest sprouting conditions. The aim of this study was to determine which temperature gives the greatest difference in seed germination and expression of seed dormancy in 10 spring wheat genotypes. The genotypes were grown in the field near Swift Current, Saskatchewan in 2000 in a randomized complete block with four replicates. Seed samples were harvested at approximately 25% moisture content (wet weight basis) and dried to 12% moisture content with minimal after-ripening. Germination was under controlled environment at temperatures of 10, 15, 20 and 30 °C in darkness. A weighted germination index (WGI) was calculated. The analysis of WGI, for each temperature, showed highly significant (p ≤ 0.01) genotype effects on germination. Most genotypes decreased in WGI (increased dormancy) as temperature was increased from 10 to 30 °C. The greatest differences in seed germination tended to be at 15 °C and 20 °C. The level of seed dormancy depended on the genotype and germination temperature. This revised version was published online in August 2006 with corrections to the Cover Date.     

The effect of gibberellic acid on male fertility in bread wheat

Summary The effect of gibberellic acid (GA₃) on male fertility was investigated in wheat. Greenhouse and field experiments were carried out. GA₃ induced high levels of male sterility both in gibberellin (GA)-sensitive and GA-insensitive genotypes. The optimum concentration was 2000 ppm of GA₃ when applied at successive sprays. The critical period for GA₃ treatment, in the general sense, extended from glume differentiation to premeiotic interphase in the oldest florets of the spike, though differences were found between GA-sensitive and GA-insensitive genotypes in the extension of this period. The effect of GA₃ on plant height was also studied. The potential use of GA₃ as a chemical hybridizing agent in wheat breeding is discussed.Abbreviations GA gibberellin - GA₃ gibberellic acid - ms male sterility Scientific Paper GEN-810 of the Instituto de Gen]'etica, CICA, INTA, Castelar, Argentina     

Functional male sterility (ps-2) in tomato (Lycopesicon esculentum Mill.) and its application in breeding and hybrid seed production

Data and findings concerning the development of an effective male sterility system (as already applied in the practice of producing tomato hybrid seed) through the use of the positional sterility (ps 2) gene are reported. Undesirable selfing and the necessity of emasculation are discussed as the two main disadvantages that limit the use of ps 2-sterile seed parents in tomato hybrid seed production. The following specific characteristics in the performance of ps 2-lines were evaluated: 1) the percentage of selfing in the ps 2-lines varies within and between the years of growing, being forever lowest during the period of hybrid seed production; 2) the percentage of selfed seeds when using a ps 2-line as seed parent in producing hybrid seed is significantly lower than the percentage of selfing, observed on this same line; 3) the ps 2-gene expressivity varies depending on the genotype. This enables the breeding of ps 2-lines exhibiting very low percentage of selfing; 4) stamen emasculation at anthesis (as applied in ps 2-lines) is significantly easier and more rapid than emasculation in floral buds. The easy maintenance of the sterile lines – by artificial selfing it is possible to produce 100% ps 2 progeny – and the high yield of hybrid seed obtained are discussed as important advantages in using this type of sterility in tomato hybrid seed production. This revised version was published online in July 2006 with corrections to the Cover Date.     

Estimation of hybrid seed production and efficient pollen flow using insect pollination of male sterile soybeans in caged plots

The aptitude of leafcutter bees to pollinate male sterile soybean plants (ms2 gene) in caged plots was evaluated in four experiments from both quantitative and qualitative points of view. The m.s. plant seed set was satisfactory: on average, it represented 60% of the male fertile isogenic seed set (with a range between 44 and 69%). The lower yield of m.s. plants was linked with a smaller number of fertile reproductive nodes. The efficient pollen flow was observed over the flowering period with both morphological and electrophoretic markers. Insect behaviour was not influenced by flower colour. Differences between flowering duration of pollen donors appear to be the major factor inducing unbalanced populations. The interest of this technique as a tool for dynamic management of the genetic variation in populations or for hybrid seed production is discussed.     

Factors responsible for levels of male sterility in photoperiod-sensitive cytoplasmic male sterile (PCMS) wheat lines

A `two-line system' using photoperiod-sensitivecytoplasmic male sterility (PCMS) caused by Aegilops crassa cytoplasm has been proposed as a newmeans of producing hybrid wheat. The PCMS line ismaintained by self-pollination under short-dayconditions (14.5 h light period), and F<sub>1</sub> seedscan be produced by outcrossing of the PCMS line witha pollinator under long-day conditions (15 h lightperiod). As the levels of male sterility in PCMSlines under the long-day conditions is a crucialfactor in determining hybrid purity of the F<sub>1</sub>seeds, a study was conducted into the effect ofseeding rate on male sterility in PCMS lines. Threedifferent density levels were tested using analloplasmic line of Japanese wheat cultivar `Norin 26'which exhibits PCMS. Levels of male sterility of thePCMS line increased at sparse planting, because tiller(ear) number per plant increased at low seedingdensity and late-appearing ears tended to exhibithigher levels of male sterility than early-appearingears. On the other hand, male sterility levels of thePCMS lines depended on genotype, e.g., the PCMS`Fujimikomugi' was completely male sterile, whereasthe PCMS `Norin 26' showed partial male sterility. APCMS line showing complete male sterility, such as thePCMS `Fujimikomugi', should produce F<sub>1</sub> seeds withhigh purity. However, the PCMS `Fujimikomugi' showeda lower female fertility. For practical use, it isnecessary to produce PCMS lines having high malesterility with high female fertility under long-dayconditions.     

Genetic and cytogenetic studies with a male sterile mutant of common wheat (Triticum aestivum)

Summary Genetic and cytogenetic studies were done on a male sterile mutant of the wheat variety Probus. Association of the 4A chromosome carrying the ms gene was studied in the F₁ of the male sterile Probus with Chinese Spring ditelo 4AS, with Transec and with line T4AS-DRS respectively. The presumption that the genetic male sterility of the mutant was due to a terminal deletion of the short arm of chromosome 4A could be confirmed.Linkage studies showed that the ms gene was at 17 map units from the dwarfing gene (Rht3) of Minister dwarf. This allows selection of short male sterile plants at the seedling stage.     

Boron deficiency induced male sterility in wheat (Triticum aestivum L.) and implications for plant breeding

Boron (B) deficiency causes grain set in wheat to fail. A wide range of genotypic variation in the response to low B has been observed. Genotypes were screened in low B in soil and sand culture, and classified into five groups, namely, very sensitive, sensitive, moderately sensitive, moderately tolerant and tolerant. At very low levels of B, the very sensitive to sensitive genotypes were completely male sterile and set only a few or no grain, while the tolerant genotypes set grain normally. Natural outcrossing was detected in these male sterile plants when a tolerant genotype was growing nearby. Grain set by cross fertilisation was markedly enhanced by a B application directly on the ear of the male sterile plants. Three practical implications are suggested. Firstly, genotypes that are tolerant to low B can provide a solution for grain set failure caused by B deficiency. Secondly, the potential for outcrossing in male sterile B deficient wheat has to be taken into account in the maintenance of pure lines in low B soils even though wheat is normally self pollinated. Thirdly, a simple and novel method for hybridization is suggested, in which B deficiency is used as fertility selective medium and male sterile female parents and fertile male parents are provided by genotypic variation in the response to low B. This revised version was published online in July 2006 with corrections to the Cover Date.     

Use of herbicide-resistant genic male sterility in hybrid rice seed production

One of the difficult problems in hybrid rice seed production is the low outcrossing frequency and requirement for much labor to produce hybrid seeds. In order to simplify the process of hybrid rice seed production, herbicide-resistant photoperiod sensitive genic male sterile (HRPGMS) rice was utilized in this study. The herbicide resistance gene bar was transferred into the photoperiod sensitive genic male sterile (PGMS) rice 920S by Agrobacterium-mediated transformation and the HRPGMS line YA3530ms with good agronomic characteristics was bred by applying conventional pedigree breeding technique. The seeds of HRPGMS and pollen parent were mixed with the ratio of 4:1 in weight, and were sowed in seedling box. The mixed seedlings of HRPGMS and pollen parent grown for 30 days were transplanted by the small transplanting machine in the field. The herbicide glufosinate ammonium was sprayed at 7 days after flowering to kill all the plants of pollen parent, whereas hybrid seeds were harvested from the survived HRPGMS parent at maturity. The outcrossing frequency of HRPGMS line from two combinations in 2002 and from five combinations in 2004 were compared with a control cultivated by the conventional 2-line system. As the result, the mean outcrossing frequency in HRPGMS of the treatments were 10.6–24.5% compared with 5.5% in PGMS of the control in 2002, and that were 24.7–32.0% compared with 7.5% in the control in 2004. Consequently, using HRPGMS in two-line system was proved to be a new method that would simplify the process of hybrid rice seed production and to increase outcrossing frequency without any artificial supplementary pollination processes.     

Effects of nucleus,cytoplasm and male sterile nucleus-cytoplasm combinations on callus initiation in anther culture of wheat

Summary Nuclear and cytoplasmic factors affect tissue culture response in wheat (Triticum aestivum), and cytoplasmic male sterility may enhance callus initiation in anther culture. Three wheat nuclear genotypes, each in normal and two alien cytoplasms conferring cytoplasmic male sterility, were evaluated for callus initiation frequency in anther culture. Nuclear genotype had the greatest effect on callus initiation, but cytoplasm and nucleus X cytoplasm interaction also produced significant effects. The nuclear genotype of Chris outperformed Butte and Coteau in all cytoplasms. Ordinary wheat and Triticum timopheevi cytoplasms outperformed Aegilops speltoides cytoplasm. Cytoplasmic male sterility did not increase callus initiation. This suggests manipulating the nuclear genotype is the best strategy for improving the capacity of wheat to initiate callus in anther culture.     

Selection for yield in wheat breeding

Summary If selection based on F₃ yield tests is to be effective, the yield tests must be successful in discriminating among yield genotypes. The available literature indicates that simple tests with limited or no replication are not very effective, although more extensive, replicated tests may be.Data from an experiment comparing F₃ yield tests with a single seed descent procedure showed that F₃ selection based on a two-replicate test with single seed descent procedure did not justify the extra work involved.     

Utilization of auxotrophy in hybrid seed production for elimination of the disadvantages of gametocids

Summary Chemically induced male sterility cannot be widely used in practice, primarily of the impractibility of regulating the dosage of gametocides under field conditions. A suboptimal dose can result in incomplete male sterility, and consequently self-seeds of the parental genotypes can contaminate the hybrid. The use of too high doses can cause not only male, but also female sterility.This problem can be eliminated through the use of recessive biochemical deficiency mutants (syn. auxotrophs). If the female plant is auxotrophic, all self-or sib-seedlings are eliminated, whereas all hybrid grains are vital.In this way, in the event of incomplete male sterility induced by gametocides, a high-value F₁ hybrid seed can be obtained by using auxotrophic mutants. This problem can also be solved in principle in the case of biparental auxotrophy, which has further advantages (blending of parents, etc.).