Allele Distribution at Locus WMS261Marking the Dwarfing Gene Rht8in Common Wheat Cultivars of Southern Ukraine

The use of codominant microsatellite molecular markers allows one to study the inheritance and distribution of alleles linked to important agronomic characters. A microsatellite locus WMS261tightly linked to a dwarfing geneRht8was analyzed in wheat cultivars and selection material of the Institute of Plant Breeding and Genetics. PCR screening of common wheat cultivars produced in the southern Ukraine showed the prevalence of a 192-bp allele at locus WMS261that indicates adaptive significance of a corresponding allele of the Rht8gene in the southern regions.     

Genetic analysis of the dwarfing gene Rht8 in wheat. Part II. The distribution and adaptive significance of allelic variants at the Rht8 locus of wheat as revealed by microsatellite screening

 Wheat microsatellite WMS 261 whose 192-bp allele has been shown to be diagnostic for the commercially important dwarfing gene Rht8 was used to screen over 100 wheat varieties to determine the worldwide spread of Rht8. The results showed Rht8 to be widespread in southern European wheats and to be present in many central European wheats including the Russian varieties ‘Avrora’, ‘Bezostaya’ and ‘Kavkaz’. Rht8 appears to be of importance to South European wheats as alternative giberellic acid (GA)-insensitive dwarfing genes do not appear to be adapted to this environment. The very successful semi-dwarf varieties bred by CIMMYT, Mexico, for distribution worldwide have been thought to carry Rht8 combined with GA-insensitive dwarfing genes. Additional height reduction would have been obtained from pleiotropic effects of the photoperiod-response gene Ppd1 that is essential to the adaptability of varieties bred for growing under short-winter days in tropical and sub-tropical areas. The microsatellite analysis showed that CIMMYT wheats lack Rht8 and carry a WMS 261 allelic variant of 165 bp that has been associated with promoting height. This presumably has adaptive significance in partly counteracting the effects of other dwarfing genes and preventing the plants being too short. Most UK, German and French wheats carry an allelic variant at the WMS 261 locus with 174 bp. This could be selected because of linkage with the recessive photoperiod-sensitive ppd1 allele that is thought to offer adaptive significance northern European wheats. Received: 17 October 1997 / Accepted: 12 November 1997     

Genetic analysis of the dwarfing gene (Rht8) in wheat. Part I. Molecular mapping of Rht8 on the short arm of chromosome 2D of bread wheat (Triticum aestivum L.)

 Two sets of single chromosome recombinant lines comparing 2D chromosomes from the wheat varieties ‘Ciano 67’ and ‘Mara’ with the common 2D chromosome of ‘Cappelle-Desprez’ in a ‘Cappelle-Desprez’ background were used to detect a diagnostic wheat microsatellite marker for the dwarfing gene Rht8. The genetic linkage maps place the wheat microsatellite marker WMS 261 0.6 cM distal to Rht8 on the short arm of chromosome 2D. By PCR analysis the WMS 261 alleles of ‘Mara’, ‘Cappelle-Desprez’ and ‘Ciano 67’ could be distinguished by different fragment sizes of 192 bp, 174 bp and 165 bp, respectively. A screen of over 100 international varieties of wheat showed that the three allelic variants were all widespread. It also demonstrated that a limited number of varieties carried novel WMS 261 variants of over 200 bp. Following classification of the individual recombinant lines for allelic variants at the WMS 261 locus it was possible to attribute a 7- to 8-cm reduction in plant height with the WMS 261-192-bp allele compared to the WMS 261-174-bp allele in the set of recombinant lines comparing 2D chromosomes of ‘Mara’ and ‘Cappelle-Desprez’. A height reduction of around 3 cm was detected between the WMS 261-174-bp allele and the WMS 261-165-bp allele in the recombinant lines comparing 2D chromosomes of ‘Cappelle-Desprez’ and ‘Ciano 67’. Received: 17 October 1997 / Accepted: 12 November 1997     

Analysis of the short-stem genes in the genotypes of bread wheat cultivars in Ukraine

Varieties of winter common wheat widely used in breeding programmes for the period of 1912-2002 in Ukraine were investigated using both the microsatellite Xgwm261 marker linked with the gibberellin-sensitive dwarf gene Rht8 and the biochemical test for the sensitivity of seedlings to gibberellic acid. Allelic characteristics of the locus Xgwm261 have been determined for 97 varieties. Near 98% of the modern varieties of winter common wheat derived from Odessa breeding institute and 50% of the varieties derived from the Mironovka wheat institute possess the allel 192 n.p. and gene Rht8 correspondingly. Estimation of sensitivity of the seedlings to gibberellic acid showed that the majority of the varieteis cultivated in Ukraine until the middle of the seventies are sensitive to gibberellic acid. The insusceptibility to gibberellic acid is predominantly manifested in the varieties derived from the south of Ukraine since the eighties owing to the introduction of the dwarf genes into the genotypes. At the same time the varieties of the Mironovka wheat institute still retain the sensitivity.     

A modern Green Revolution gene for reduced height in wheat

Increases in the yield of wheat during the Green Revolution of the late 20^th century were achieved through the introduction of Reduced height (Rht) dwarfing genes. The Rht‐B1 and Rht‐D1 loci ensured short stature by limiting the response to the growth‐promoting hormone gibberellin, and are now widespread through international breeding programs. Despite this advantage, interference with the plant's response to gibberellin also triggers adverse effects for a range of important agronomic traits, and consequently modern Green Revolution genes are urgently required. In this study, we revisited the genetic control of wheat height using an association mapping approach and a large panel of 1110 worldwide winter wheat cultivars. This led to the identification of a major Rht locus on chromosome 6A, Rht24, which substantially reduces plant height alone as well as in combination with Rht‐1b alleles. Remarkably, behind Rht‐D1, Rht24 was the second most important locus for reduced height, explaining 15.0% of the genotypic variance and exerting an allele substitution effect of –8.8 cm. Unlike the two Rht‐1b alleles, plants carrying Rht24 remain sensitive to gibberellic acid treatment. Rht24 appears in breeding programs from all countries of origin investigated, with increased frequency over the last decades, indicating that wheat breeders have actively selected for this locus. Taken together, this study reveals Rht24 as an important Rht gene of commercial relevance in worldwide wheat breeding.     

The effects of gene Rht8 alleles in agronomic traits of winter bread wheat in the conditions of South Steppe of the Ukraine

Resistance to frost, winterhardiness, yield and its components were investigated in recombinant-inbred lines F5 Odesskaya 16/Bezostaya 1 and 61 winter wheat varieties differing on the alleles of Rht8 gene. The absence of influence of gene Rht8 different alleles on the tested traits was shown. The high frequency of distribution of alleles Rht8c in modern wheat varieties developed in Plant Breeding and Genetics Institute is a result of high frequency of use of this allele in genetic pool after 1959.     

Effects of the alleles of the <Emphasis Type="Italic">Rht8</Emphasis> gene on the agricultural traits of bread winter wheat under conditions of the steppe region of the Southern Ukraine

Resistance to frost and winter hardiness, as well as crop capacity and its components, were studied in the recombinant-inbred F₅ Odesskaya and 16/Bezostayal lines and also in 61 lines of winter wheat carrying different alleles of the Rht8 gene. We observed no influence of different alleles of the Rht8 gene on the traits under consideration. The preponderance of the Rht8c allele in modern wheat lines developed at the Plant Breeding and Genetics Institute is therefore a result of its being the most frequently used allele in the genetic pool since 1959.     

矮秆基因对小麦部分农艺性状的效应

以中国主要麦区的124份小麦品种为材料,利用分子标记和系谱分析相结合,对其按照所含的矮秆基因Rht-B1b、Rht-D1b和Rht8进行分类,结合田间株高、旗叶长、小穗数和穗粒数以及室内苗期根系长度等农艺形状的调查,分析不同矮秆基因对小麦农艺性状的效应.结果显示:(1)参试的124份小麦品种(系)中23份含有Rht-B1b,7份含有Rht-D1b,22份含有Rht8基因,34份同时含有Rht-B1b和Rht8,16份同时含有Rht-D1b和Rht8,可分为6组.(2)Rht-B1b和Rht-D1b在降低株高的同时也缩短了旗叶的长度和苗期叶长,Rht8对株高的影响较弱,对旗叶和苗期叶长的影响也较小;3个矮秆基因对苗期根系长度、小穗数没有显著影响;Rht-D1b和Rht8显著增加穗粒数.研究表明,矮秆基因Rht8对小麦株高以及其他农艺性状的影响均较小,但能够显著增加穗粒数,是小麦矮化育种中比较理想的矮秆基因.     

Microsatellite Marker Linked to a Leaf Rust Resistance Gene from Triticum monococcum L Transferred to Bread Wheat

Triticum monococcum L, a diploid wheat species closely related to the A genome of cultivated wheats, is highly resistant to leaf rust. A synthetic amphiploid, T. monococcum — T. durum was crossed with T. aestivum cv WL711, highly susceptible to leaf rust. Leaf rust resistant derivatives were selected among backcross generations with the recurrent parent WL711 and cytologically analysed. Chromosome number of the leaf rust resistant BC₁F₃ progenies varied from 39 to 44. Six leaf rust resistant and susceptible bulks from different BC₁F₃ progenies were analysed using 29 wheat microsatellite(WMS) markers already mapped on A genome of bread wheat and found polymorphic among parents. One T. monococcum specific allele of WMS gwm136 locus was found to be closely linked to the leaf rust resistance gene in all the resistant bulks. Differential chromosome number, frequency of univalents and multivalents, however, indicated that the critical T. monococcum chromosome might be present in addition to the A genome chromosomes of wheat, substituted either for the B or D genome chromosome of wheat or translocated to chromosome 1A of wheat in one or the other bulks. The association of the T. monococcum specific allele of WMS gwm136 locus to leaf rust resistance was further confirmed from bulked segregant analysis in BC₂F₁ generation.     

A new reduced height gene found in the tetraploid semi-dwarf wheat landrace Aiganfanmai

Aiganfanmai is a dwarf tetraploid wheat landrace (Triticum turgidum var. turgidum) that stably produces the semi-dwarf trait. Plant height varies from 80-105 cm under cultivation. Compared with tall durum wheat (T. turgidum var. durum) variety Langdon, we found it to have short spikes and seeds, besides a semi-dwarf character. We crossed Aiganfanmai with Langdon to analyze the genetic basis of the semi-dwarf trait. The F(2) population segregated at a 1:3 ratio for the short trait to the normal, which demonstrates that Aiganfanmai carries a recessive reduced height (Rht) gene. This gene was found to be located between the molecular markers Xgwm471 and Xgwm350 on chromosome arm 7AS by microsatellite analysis. No Rht gene had been reported from this chromosome; we designated it as Rht22. Rht 22, unlike other previously reported Rht genes, does not reduce internodal cell length. Reduced cell numbers might explain the short stem trait.     

Characterization of a Putative New Semi-Dominant Reduced Height Gene,Rht_NM9, in Wheat(Triticum aestivum L.)

Plant height is an important agronomic trait in cereal crops, and can affect both plant architecture and grain yield. New dwarfing genes are required for improving the genetic diversity of wheat. In this study, a novel dwarf mutant, NM9, was created by treating seeds of the wheat variety NAU9918 with ethyl methanesulfonate(EMS). NM9 showed obvious phenotypic changes, which were distinct from those caused by other dwarfing genes, especially the reduced plant height, increased effective tiller number, and elongated spike and grain length. The reduced plant height in NM9 was attributable to a semi-dominant dwarfing gene Rht_NM9, which was flanked by two closely linked SNP markers, SNP34 and SNP41, covering an 8.86-Mb region on the chromosome arm 2AS. The results of gibberellic acid(GA) sensitivity evaluation, comparative genomics analysis and allelism test indicated that Rht_NM9 was neither allelic to Rht7 and Rht21 nor homoeoallelic to Rht8, so Rht_NM9 was proposed to be a new dwarfing locus on the homoeologous group 2 chromosomes of wheat. Rht_NM9has a negative effect on plant height and positive effects on effective tiller number and grain size, thus, Rht_NM9 could be used for elucidating the mechanisms underlying plant architecture and grain development.     

Microsatellite mapping of the powdery mildew resistance gene <Emphasis Type="Italic">Pm5e</Emphasis> in common wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.)

Powdery mildew, caused by Erysiphe graminis DM f. sp. tritici (Em. Marchal), is one of the most important diseases of common wheat world-wide. Chinese wheat variety 'Fuzhuang 30' carries the powdery mildew resistance gene Pm5e and has proven to be a valuable resistance source of powdery mildew for wheat breeding. Microsatellite markers were employed to identify the gene Pm5e in a F(2) progeny from the cross 'Nongda 15' (susceptible) x 'Fuzhuang 30' (resistant). The gene Pm5e was mapped in the distal region of chromosome 7BL. Seven microsatellite markers were found to be linked to the gene Pm5e, of which two codominant markers Xgwm783 and Xgwm1267 were relatively close to Pm5e with a linkage distance of 11.0 cM and 6.6 cM, respectively. It is possible to use the 136-bp allele of Xgwm1267 in 'Fuzhuang 30' for marker-assisted selection during the wheat resistance breeding process for facilitation of gene pyramiding. The mapping information in the present study provides a starting point for fine mapping of the Pm5 locus and map-based cloning to clarify the molecular structure and function of the different alleles at the Pm5 locus. A microsatellite linkage map of chromosome 7B was constructed with 20 microsatellite loci, nine on the short arm and 11 on the long arm. This information will be very useful for further mapping of agronomically important genes of interest on chromosome 7B.     

Effect of the Rht3 dwarfing gene on dynamics of cell extension in wheat leaves, and its modification by gibberellic acid and paclobutrazol

The second leaf of wheat was used as a model system to examinethe effects of the Rht3 dwarfing gene on leaf growth. Comparedto the rht3 wild type, the Rht3allele decreased final leaf length,surface area and dry mass by reducing the maximum growth rates,but without affecting growth duration. Gibberellic acid (GA₃)increased final leaf length and maximum growth rate in the rht3wild type, but was without effect on the Rht3 mutant, whichis generally regarded as being non-responsive to gibberellin(GA). Paclobutrazol, an inhibitor of GA biosynthesis, decreasedfinal leaf length and maximum growth rate in the rht3 wild typeto values similar to those in the untreated Rht3 mutant. NeitherGA₃ nor paclobutrazol affected the duration of leaf growth.The decrease in leaf length was produced by reduction of celllength rather than cell number. The maximum relative elementalgrowth rate (REGR) for cell extension was essentially the samein all treatments, as was the time between the cells leavingthe meristem and achieving maximum extension rate. The differencesbetween the genotypes and treatments were all almost entirelydue to differences in the time taken from the attainment ofmaximum REGR of cell extension to the cessation of extension.This was reflected in the length of the extension zone, whichwas approximately 6–8 per cent of final leaf length. Theeffects of the Rht3 allele, GA₃ and paclobutrazol all appearto be on the processes which promote the cessation of cell elongation. Key words: Cell extension, gibberellin, leaf growth, Rht3 gene, Triticum, wheat     

Mapping a resistance gene in wheat cultivar Yangfu 9311 to yellow mosaic virus, using microsatellite markers

Wheat yellow mosaic disease, which is caused by wheat yellow mosaic bymovirus (WYMV) and transmitted by soil-borne fungus, results in severe damage on wheat (Triticum aestivum L.) production in China. For development of resistant cultivars to reduce wheat yield losses due to wheat yellow mosaic disease, resistance test and genetic analysis indicated that a single dominant gene in wheat cultivar Yangfu 9311 contributed to the resistance. Bulk segregant analysis was used to identify microsatellite markers linked to the resistance gene in an F₂ population derived from the cross Yangfu 9311 (resistant) × Yangmai 10 (susceptible). Microsatellite markers Xwmc41, Xwmc181, Xpsp3039, and Xgwm349 were co-dominantly or dominantly linked with the gene responsible for WYMV resistance at a distance of 8.1–11.6 cM. Based on the wheat microsatellite consensus map and the results from amplification of the cultivar Chinese Spring nulli-tetrasomic stocks, the resistance gene to wheat yellow mosaic disease derived from Yangfu 9311, temporarily named as YmYF, was thus mapped on the long arm of chromosome 2D (2DL).     

Microsatellite markers reveal chimeric origin of redesignated chromosome 4A of wheat from Triticum urartu and other species.

Although a new nomenclature has been adopted for wheat in which chromosome 4A (4AO) has been renamed 4B (4BN) and chromosome 4B (4BO) has been renamed 4A (4AN), their specific origin remains uncertain. The use of wheat microsatellite (WMS) markers mapped to chromosomes 4AN and 4BN in a set of polyploid wheats and diploid genome donors has unequivocally indicated that the entire short arm of 4AN, some part of 4ANL. and a segment of 4BNL were derived from Triticum urartu. The presence of a T. urartu-specific allele at locus gwm368 on 4BNL and of an Aegilops speltoides allele at locus gwm397 on 4ANL suggests the possibility of a reciprocal translocation between 4ANL and 4BNL. The subcentromeric and telomeric regions of 4ANL corresponding to heterochromatic C-bands were derived neither from diploid wheats nor from Ae. speltoides or Aegilops longissima.     

New data on the distribution of hybrid necrosis genes in winter bread wheat (Triticum aestivum L.) cultivars

Hybrid necrosis genotypes have been identified in 125 Russian cultivars of winter bread wheat. More than half of them (56%) carry the Ne2 gene (genotype ne1ne1Ne2Ne2); others are free of necrosis genes (genotype ne1ne1ne2ne2). The possible causes of the increase in the Ne2 allele frequency and the loss of the Ne1Ne1ne2ne2 genotype in modem Russian cultivars of winter wheat are discussed. The principal component method has been used to compare the structures of the genetic diversity of cultivars differing in the hybrid necrosis genotype. It has been found that the Ne2 allele in winter wheat cultivars from northern Russia has originated from the cultivar Mironovskaya 808, whereas the cultivar Bezostaya 1 is not a source of this gene. In cultivars from southern Russia, the presence of the Ne2 allele is also mainly accounted for by the use of Mironovskaya 808 wheat in their breeding. The recessive genotype is explained by the presence of descendants of the cultivar Odesskaya 16 in the pedigrees of southern Russian winter wheats. The genetic relationship of cultivars with identical and different necrosis genotypes has been analyzed in nine regions of the Russian Federation.     

一个来自硬粒小麦的抗白粉病基因的鉴定和微卫星标记

在起源于硬粒小麦(Triticum durum Desf.accession DR147)和尾状山羊草(Aegilops caudata L.acc.Ae14)合成的双二倍体与普通小麦品种"莱州953"杂交组合衍生的BC3F2群体中鉴定了一个抗小麦白粉病基因.遗传分析表明,该基因为一个显性单基因.应用分离群体分组法(BSA),鉴定了两个与抗病基因紧密连锁的微卫星标记Xgwm311和Xgwm382,它们与抗病基因的遗传距离分别为5.9 cM和4.9 cM.对双二倍体亲本硬粒小麦DR147和尾状山羊草Ae14及轮回亲本"莱州953"的DNA PCR扩增结果表明,与抗病基因相关的微卫星标记Xgwm311和Xgwm382来源于硬粒小麦DR147.根据已发表的小麦微卫星图谱和对"中国春"缺-四体系DNA扩增结果,抗病基因被定位在小麦2A染色体的长臂末端.     

Polymorphism of cattle prolactin gene: microsatellites, PSR-RFLP]

In the samples of Russian Ayrshire and Gorbatov Red cattle breeds, distribution of frequencies of prolactin (PRL) gene alleles generated due to the presence of polymorphic RsaI site in exon 3 were studied. In the breeds, the frequencies of the B allele of the PRL gene (with RsaI(+) site) detected by the PCR-RFLP method were 14.1 and 8.6%, respectively. In Black Pied, Ayrshire and Gorbatov Red cattle breeds, variation of the microsatellite dinucleotide repeat in the regulatory region of the gene PRL was also studied. Gorbatov Red breed was monomorphic at the microsatellite locus with the only allele 164 bp in length. Two alleles (164 bp and 162 bp) were detected in the other breeds studied. The frequencies of 164-bp allele of the microsatellite locus were 93.7 and 90.0% in Black Pied and Ayrshire breeds, respectively. In Gorbatov Red breed of dairy type with good beef qualities and low milk-fat yield, lower level of heterozygosity for PRL gene was demonstrated compared to Ayrshire and Black Pied breeds with high milk-fat yield. In three cattle breeds, higher mean estimate of polymorphism information content of PCR-RFLP in exon 3 (PIC = 0.21) was revealed compared with the same estimate (PIC = 0.09) for the microsatellite locus variability in the regulatory region of the PRL gene. Characteristics of allele B distribution of the PRL gene in the representatives of the Bovidae family are considered.     

Sequence tagged microsatellites for the Xgwm533 locus provide new diagnostic markers to select for the presence of stem rust resistance gene Sr2 in bread wheat ( Triticum aestivum L.)

The stem rust resistance gene Sr2 has provided durable broad-spectrum, adult-plant resistance to the fungal pathogen Puccinia graminis Pers. f. sp. tritici throughout wheat-growing regions of the world for more than 50 years. The ability to select for Sr2 in wheat breeding programs was recently improved by the identification of a tightly linked microsatellite marker gwm533. This marker typically amplifies a 120-bp polymerase chain reaction fragment from wheat lines carrying Sr2. In instances where the 120-bp fragment is not associated with the presence of Sr2, DNA sequence analysis has shown that a second allele was amplified, differing in the structure of the microsatellite repeat. To discriminate this allelic homoplasy (alleles identical in size, but not identical by descent), sequence-tagged microsatellites (STM) markers were developed for the Xgwm533 locus. These markers were shown to be diagnostic for the presence of Sr2 in a wide range of germplasm, representative of all major wheat varieties historically grown in Australia. The STMs will be particularly useful for marker-assisted selection in Southern Australian breeding programs, where the use of the marker gwm533 is often precluded by the presence of the non-Sr2-associated 120-bp allele in the pedigree of current breeding germplasm. The STMs also revealed a high incidence of previously undetected allelic homoplasy at the Xgwm533 locus and may have broader utility in genetic research and breeding, as this locus is also reported to be strongly associated with a major gene conferring resistance to Fusarium head blight.