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.     

A Dominant Gene for Male Sterility in Wheat

A dominant gene inducing male sterility in wheat is described. It is located on the short arm of chromosme 4D with a recombination percentage of 31.16 with the centromere. The potential use of this allele in breeding and cytogenetic studies in both tetraploid and hexaploid with is discussed.     

An EMS-induced wheat mutant restoring fertility against photoperiod-sensitive cytoplasmic male sterility

Triticum aestivum cv. 'Norin 26' with Aegilops crassa cytoplasm shows photoperiod-sensitive cytoplasmic male sterility (PCMS). This alloplasmic line is almost completely male-sterile under long-day conditions (≥ 15h), but highly male-fertile under short-day conditions (≤ 14.5h). To obtain male—fertile mutants against PCMS, seeds of the alloplasmic line were treated with ethyl methane sulfonate (EMS). The M₃ generation was evaluated for PCMS expression, and one fertility-restoring (FR-mutant) line showing high male fertility under the long-day conditions was selected. Reciprocal F, hybrids between the FR-mutant and the alloplasmic 'Norin 26' showed male sterility under the long-day conditions, and continuous segregation with respect to the degree of fertility restoration occurred in their F₂ generations. These results indicate that multiple recessive mutations with minor effects, induced in the nuclear genome, are involved in the fertility restoration. In fact, no restriction-fragment-length polymorphisms of mitochondrial DNA between the FR-mutant and the alloplasmic 'Norin 26' are found.     

Genetic diversity among populations and breeding lines from recurrent selection in Brassica napus as revealed by RAPD markers

Recurrent selection facilitated by dominant male sterility has been conducted to broaden the genetic basis for cultivar development in Brassica napus. This study aimed to evaluate the genetic variation in four base populations (C0‐C3) and breeding lines from two of the populations produced during recurrent selection by random amplified polymorphic DNA (Rapd) markers. Genetic variation in four populations declined gradually with the advance of selection cycles as measured by expected genetic heterozygosity (from 0.2058 in C0 to 0.1536 in C3) but the decline was not statistically significant. When compared with the average genetic distances for 21 germplasm collections with wide geographical and genetic origins (0.4712) and seven breeding lines from pedigree selection (0.2059), seven breeding lines selected from the C1 population and 11 from the C3 population had a larger average genetic distance (0.5339 and 0.5486, respectively). Clustering analysis indicated that the lines from recurrent selection had a much lower genetic similarity than lines from pedigree selection. Our results suggest that base populations derived from recurrent selection could provide a wider genetic variation for selection of breeding lines with more broad genetic bases.     

Inheritance and chromosome location of photoperiod‐thermo sensitive male sterility in wheat line Xinong 291S

The present research aimed to study the inheritance and chromosomal location of a photoperiod‐thermo sensitive male sterility (PTSMS) gene in Xinong 291S (XN291S), which is a new PTSMS wheat line. The inheritance was studied in F₁ and F₂ populations derived from crosses between XN291S and eight wheat cultivars. All F₁ plants were fertile and the F₂ populations segregated in either 15 : 1 or 3 : 1 fertile : sterile ratios indicating that PTSMS was controlled by one or two recessive major genes. Five wheat cultivars carried homozygous dominant alleles for fertility, whereas the other three, including ‘Chinese Spring’ (CS), carried a single homozygous dominant allele. Chromosomal location of the PTSMS gene was studied by crossing a set of CS nulli‐tetrasomic lines to XN291S. Self‐fertility of F₁ hybrids XN291S/N5BT5A and XN291S/N5BT5D were significantly lower than the others. Therefore, the PTSMS gene in XN291S that differed from CS was located on chromosome 5B. Location of the second PTSMS gene needs further study.     

Genetic analysis of fertility restoration against photoperiod-sensitive cytoplasmic male sterility in Triticum aestivum cv. Norin 61

Triticum aestivum cv. Norin 26 with the Aegilops crassa cytoplasm becomes almost completely male sterile when grown under a long-day condition (15 h of light or longer), but is highly male fertile under a short-day condition (14.5h or less). This type of male sterility is called photoperiod-sensitive cytoplasmic male sterility (PCMS). Genetic analyses were made of the fertility-restoring (Rf) genes effective against PCMS that are present in T. aestivum cv. Norin 61. Conventional and monosomic studies indicated that restoration of fertility is controlled by multiple Rf genes located on at least four chromosomes: 4 A, 1D, 3D and 5D. The genetic mechanism of fertility restoration by the genes of‘Norin 61’differs from the mechanisms reported for‘Chinese Spring’and a‘Norin 26’mutant line.     

Ethrel-a male gametocide that can replace the male sterility genes in barley

Summary The use of ethrel for hand crossing in recurrent selection programs with barley is advocated.     

Molecular markers and their applications in wheat breeding

In recent years, considerable emphasis has been placed on the development of molecular markers to be used for a variety of objectives. This review attempts to give an account of different molecular markers—restriction fragment length polymorphisms (RFLPs), random amplified polymorphic DNAs (RAPDs), sequence-tagged sites (STS), DNA amplification fingerprinting (DAF), amplified fragment length polymorphisms (AFLPs) and microsatellites (STMS)—currently available for genome mapping and for tagging different traits in wheat. Other markers, including microsatellite-primed polymerase chain reaction (MP-PCR), expressed sequence tags (ESTs) and single nucleotide polymorphisms (SNPs) are also discussed. Recent information on synteny in cereal genomes, marker-assisted selection, marker validation and their relevance to cereal breeding in general and wheat breeding in particular are also examined.     

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.     

Improving male fertility restoration of common wheat for Triticum timopheevii cytoplasm

The fertile pure line R3‐37 of common wheat with cytoplasm of Triticum timopheevii Zhuk. is an R‐line (restorer) that can restore the male fertility of A‐lines (male sterile lines) with T. timopheevii cytoplasm. In breeding hybrid wheat, the hybrid of the cross R3‐37/ Baimian3 was found to be completely male sterile, indicating that Baimian3 has some genes that are epistatic to the Rf genes in R3‐37. In order to elucidate the essence of this phenomenon, the male fertilities of the hybrids of 27 crosses including R3‐37 and/or Baimian3 were studied. The results show that inheritance of male fertility of the hybrid R3‐37/Baimian3 involves interactions among Rf alleles, male fertility‐inhibiting genes and genetic background. Although more than 70 different kinds of male sterile cytoplasm to common wheat have been discovered, the systems of hybrid wheat production based on male sterile cytoplasm are all the A‐line/R‐line type and all have similar problems of hybrid fertility restoration. This study confirmed that there is a new model (A‐line/R*‐line//R‐line) for producing hybrid wheat with high fertility restoration. In the new model, the completely male sterile hybrids of A‐line/R*‐line can act as common A‐line.     

黄淮麦区Fhb1基因的育种应用

小麦赤霉病是由禾谷镰孢菌引起的一种世界性重要病害,严重威胁小麦生产安全。黄淮麦区作为我国小麦主产区,赤霉病危害日趋严重,因缺乏半冬性抗源,抗赤霉病育种进展缓慢。Fhb1基因是迄今发现的效应最大、抗性最稳定,也是被广泛应用于全球小麦赤霉病抗性育种的主效基因,但Fhb1基因在黄淮麦区尚未被广泛应用。本研究以感病品种矮抗58为轮回亲本, H35为Fhb1基因供体亲本,通过有限回交和分子标记辅助选择,同时利用双单倍体育种和传统系谱选育两种方法,培育出了一批综合性状较好、具有Fhb1基因的优良新品系,其中徐麦DH9和徐麦17252经多年鉴定均达到中抗水平。在以徐麦36和徐麦2023为杂交父本的后代品系中,含Fhb1基因的家系赤霉病平均抗性明显优于感病对照。Fhb1基因的导入显著提高了赤霉病抗性,但部分家系对赤霉病仍旧表现出高感水平,说明赤霉病抗性还受到Fhb1基因以外其他遗传因素的显著影响。本研究为Fhb1基因在黄淮麦区抗赤霉病小麦育种中的应用提供了成功的经验。     

Development of dominant nuclear male-sterile lines with a blue seed marker in durum and common wheat

In order to develop genie male‐sterile lines with a blue seed marker, male‐sterile plants, controlled by a dominant nuclear gene Ms2, were used as female parents against a 4E disomic addition line ‘Xiaoyan Lanli’(2n= 44, AABBDD+4EII) as the male parent to produce monosomic addition lines with blue seed. Male‐sterile plants from the monosomic addition lines were pollinated with durum wheat for several generations and in 1989 a male‐sterile line with the blue grain gene and the male‐sterile gene Ms2 on the same additional chromosome was detected and named line 89‐2343. Using this line, the blue seed marker was successfully added to a short male‐sterile line containing Ms2 and Rht10. The segregation ratios of male sterility and seed colour as well as the chromosome figurations of different plants indicated that the blue grain genes, Ms2 and Rht10 were located on the same additional chromosome. Cytological analysis showed that the blue marker male‐sterile lines in durum wheat and common wheat were monosomic with an additional chromosome 4E. The inheritance ratio for blue seed male‐sterile plants and white seed male‐fertile plants was 19.7% and 80.3%, respectively, in common wheat. The potential for using blue marker sterile lines in population improvement and hybrid production is discussed.     

SSR and SCAR markers linked to the fertility-restoring gene for a D2-type cytoplasmic male-sterile line in wheat

‘Yi 4060’ is an elite restorer line of a non‐photoperiod‐sensitive D²‐type cytoplasmic male‐sterile (CMS) line of wheat. Random amplified polymorphic DNA (RAPD) and simple sequence repeat (SSR) markers were employed to map one major fertility‐restoring gene (D²Rf₁) in ‘Yi 4060′. The sterile and fertile DNA pools were established from individuals in BC₆, based on bulked segregant analysis. One RAPD marker E09, linked to D²Rf₁, was converted to a SCAR marker and designated as E09‐SCAR₈₆₅. The genetic distance between E09‐SCAR₈₆₅ and D²Rf₁ is 9.5 cM. Two SSR markers, Xgwm11 and Xgwm18, were also linked to a D²Rf₁ and co‐segregated with E09‐SCAR₈₆₅. The three molecular markers are useful in marker‐assisted breeding of the elite restorer lines for D² ‐type CMS lines in wheat.     

Comparison of two fertility restoration systems against photoperiod-sensitive cytoplasmic male sterility in wheat

A ‘two‐line system’ using photoperiod‐sensitive cytoplasmic male sterility (PCMS) caused by Aegilops crassa cytoplasm under a long‐day photoperiod ( 15 h) has been proposed as a new means of producing hybrid varieties in common wheat. 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 under long‐day conditions. Two kinds of fertility restoration systems against the PCMS are known. One is involved with a set of multiple fertility‐restoring (Rf) genes in the wheat cultivar ‘Norin 61’ located on (at least) chromosomes 4A, 1D, 3D and 5D. The other is controlled by a single dominant major Rf gene, Rfd1, located on the long arm of chromosome 7B in the wheat cultivar ‘Chinese Spring’. To examine the degree of fertility restoration by these two systems, nine PCMS lines were crossed with ‘Norin 61’ and ‘Chinese Spring’ as the restorer lines, and the F₁ hybrids were investigated. The degree of fertility restoration was estimated by comparing the seed set rates in the F₁ hybrids having the Ae. crassa cytoplasm and those with normal cytoplasm. The results revealed that the fertility restoration ability of a set of multiple Rf genes in ‘Norin 61’ was higher than that of the Rfd1 gene in ‘Chinese Spring’.     

Variation in sterility among wheat (Triticum aestivum L.) genotypes in response to boron deficiency in Nepal

Two field experiments were carried out at the Khairanitar Farm, Nepal during the winter season of 1993 to investigate whether wheat genotypes differ in their sterility response to low soil boron and whether boron added to soil can correct sterility. A nursery consisting of 41 diverse genotypes from Nepal, India, Bangladesh, Thailand, China, Pakistan and CIMMYT/Mexico were evaluated in a plot where severe sterility had been observed in previous years. The second experiment involved six genotypes of known sterility responses to boron grown with or without an application of boron at 1 kg ha^-1. Large differences were observed among genotypes in terms of number of grain set per ear (range 0.5 to 30 ) and sterility (5.5% to 97.5%). About one third of all the genotypes had <25% sterility and 16 of them suffered >75% sterility. The genotypes Fang-60, Sonalika, De Mai 6-22, BUC/FLK/MYNA/VL and HDW-234 were highly tolerant to boron deficiency with <10% sterility. In sharp contrast, genotypes BOW/BUT, SERI/THB, Glennson, SW-41, Yunmai-33 and UHU were highly susceptible and set virtually no grains in the boron deficient plots. Strong responses to boron application were observed and genotypic variations were evident. Boron at 1 kg ha^-1significantly reduced the number of late ears, increased number of grains per ear and grain yield in boron-responsive genotypes. Tolerant genotypes were not influenced by added boron. Given these striking genetic differences, progress in breeding wheat for B-deficient soils in Nepal should be worthwhile. This revised version was published online in July 2006 with corrections to the Cover Date.     

Ethyloxanilates as specific male gametocides for wheat (Triticum aestivum L.)

Induction of male sterility by deployment of male gametocides holds immense potential in heterosis breeding of wheat. The efficacy of a new class of male gametocide for wheat (Triticum aestivum L.) is described: ethyloxanilates, the most active example of this class being ethyl 4‐fluorooxanilate (E4FO). E4FO induces male sterility, specifically, without detectable effects on various agronomic features and female fertility. The plants sprayed once with 0.15% E4FO exhibited 100% pollen and floret sterility without causing a significant reduction in total yield. E4FO was screened on 29 genotypes of wheat at 0.15% test concentration and was observed to induce 99.76 ± 0.37% male sterility. Thirteen F₁‐hybrids of wheat were produced using the gametocide in Winter 2000–2001 and were evaluated for their agronomic performance in Winter 2001–2002. The cross combinations viz., lines WR 544 × HW 2046 and HW 2044 × WR 956 have outperformed their respective better parents by 48.17% and 23.42% in grain yield/plant and thus have potential as hybrids.     

Molecular mapping of a dominant genic male sterility gene Ms in rapeseed (Brassica napus)

Rs1046AB is a genic male sterile two‐type line in rapeseed that has great potential for hybrid seed production. The sterility of this line is conditioned by the interaction of two genes, i.e. the dominant genic male sterility gene (Ms) and the suppressor gene (Rf). The present study was undertaken to identify DNA markers for the Ms locus in a BC₁ population developed from a cross between a male‐sterile plant in Rs1046AB and the fertile canola‐type cultivar ‘Samourai’. Bulked segregant analysis was performed using the amplified fragment length polymorphism (AFLP) methodology. From the survey of 480 AFLP primer combinations, five AFLP markers (P10M13₃₅₀, P13M8₄₀₀, P6M6₄₁₀, E7M1₂₃₀ and E3M15₁₀₀) tightly linked to the target gene were identified. Two of them, E3M15₁₀₀ and P6M6₄₁₀, located the closest, at either side of Ms at a distance of 3.7 and 5.9 cM, respectively. The Ms locus was subsequently mapped on linkage group LG10 in the map developed in this laboratory, adding two additional markers weakly linked to it. This suite of markers will be valuable in designing a marker‐assisted genic male sterility three‐line breeding programme.     

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     

A selection tool for freezing tolerance in common wheat using the fast chlorophyll a fluorescence transient

The analysis of fast chlorophyll fluorescence induction kinetics in dark-adapted leaves (JIP-test) is proposed as a rapid method for evaluating freezing tolerance in wheat. Wheat leaves were detached from the plants of 40 cultivars grown in an open-field in Smolice, during December and March of winters 2004/2005 and 2005/2006, transferred to laboratory in Krakow and frozen in −15°C. Next, measurements of chlorophyll fluorescence were taken. The correlation coefficients between the JIP-test parameters with freezing tolerance data obtained in multiple field-laboratory studies, varied between 0.58 and 0.70 depending on sampling time and parameter measured. The chlorophyll fluorescence parameter, which can always be used for the screening of freezing tolerance in the method described here, is the performance index (PI), which characterizes the overall energy flow efficiency in photosystem II (PSII). The results indicate, that the technique proposed here can be useful for freezing tolerance screening in plant breeding programmes. However, this method seems to be a more reliable tool in the selection of freezing tolerant germplasm, than for discarding freezing susceptible materials.     

Cytoplasmic male sterility in Brassicaceae crops

Brassicaceae crops display strong hybrid vigor, and have long been subject to F₁ hybrid breeding. Because the most reliable system of F₁ seed production is based on cytoplasmic male sterility (CMS), various types of CMS have been developed and adopted in practice to breed Brassicaceae oil seed and vegetable crops. CMS is a maternally inherited trait encoded in the mitochondrial genome, and the male sterile phenotype arises as a result of interaction of a mitochondrial CMS gene and a nuclear fertility restoring (Rf) gene. Therefore, CMS has been intensively investigated for gaining basic insights into molecular aspects of nuclear-mitochondrial genome interactions and for practical applications in plant breeding. Several CMS genes have been identified by molecular genetic studies, including Ogura CMS from Japanese radish, which is the most extensively studied and most widely used. In this review, we discuss Ogura CMS, and other CMS systems, and the causal mitochondrial genes for CMS. Studies on nuclear Rf genes and the cytoplasmic effects of alien cytoplasm on general crop performance are also reviewed. Finally, some of the unresolved questions about CMS are highlighted.