Identification and chromosomal locations of aconitase gene loci in Triticeae species

Summary Two systems of monomeric aconitase (ACO) isozymes, designated ACO-1 and ACO-2, were identified in Triticum aestivum and in five diploid Triticeae species. The gene loci Aco-A1, Aco-B1, and Aco-D1 were located in T. aestivum cv. Chinese Spring chromosome arms 6Aq, 6Bq, and 6Dq, respectively, and the gene loci Aco-A2, Aco-B2, and Aco-D2 in 5 Aq, 5 Bq, and 5Dq, respectively. Aco-1 gene loci were also identified in 6E of Elytrigia elongata, 6HL of Hordeum vulgare cv. Betzes, 6RL of Secale cereale PI 252003, 6S¹ of T. longissimum, and CSU-31 of T. umbellulatum. Other Aco-2 gene loci were identified in 5RL of S. cereale cv. King II and 4EL of E. elongata. Conservation of synteny relationships is indicated among the species studied for the genes identified, with the exception of Aco-E2; the presence of this gene in 4EL suggests that E. elongata differs from Chinese Spring and King II by a translocation involving 4E and 5E.Adapted from a thesis submitted to the Graduate College, Texas A&M University, by K.J.C. in partial fulfillment of the requirements for the M.S. degree in Genetics     

Genetic analysis of Triticeae shikimate dehydrogenase

Starch gel electrophoresis and polyacrylamide gel isoelectric focusing (IEF) were used to investigate the genetic control of Triticeae shikimate dehydrogenase-1 (SKDH-1). Studies of wheat-alien species chromosome addition lines established thatSkdh-1 ofHordeum vulgare cv. Betzes is located in chromosome 5H,Skdh-V1 ofDasypyrum villosum in 5V,Skdh-R1 ofSecale cereale cvs. Dakold and King II in 5R, andSkdh-S ¹1 ofTriticum longissimum in 5S¹S. Also, the chromosomal locations of the genes that encode SKDH-1 inT. aestivum cv. Chinese Spring,T. umbellulatum, andS. cereale cv. Imperial, determined earlier using zone electrophoresis, were reconfirmed using IEF. Zone electrophoresis and IEF do not differ markedly in their ability to detect the expression of alienSkdh-1 genes in wheat-alien species chromosome addition lines. However, IEF may be superior to zone electrophoresis as a technique for detecting and analyzing SKDH-1 genetic variants within Triticeae species; among the species studied, IEF generally resolved two or more isozymes perSkdh-1 allele present, while zone electrophoresis resolved only one.Technical article No. 22791 of the Texas Agricultural Experiment Station. This paper is based upon research supported in part by the U.S. Department of Agriculture under Agreement No. 83-CRCR-1-1322.     

Molecular characterization of the genome composition of partial amphiploids derived from Triticum aestivum×Thinopyrum ponticum and T. aestivum×Th. intermedium as sources of resistance to wheat streak mosaic virus and its vector, Aceria tosichella

 Wheat streak mosaic virus (WSMV), vectored by the wheat curl mite (WCM), is one of the most important viral diseases of wheat (Triticum aestivum) in the world. Genetic resistance to WSMV and the WCM does not exist in wheat. Resistance to WSMV and the WCM was evaluated in five different partial amphiploids namely Agrotana, OK7211542, ORRPX, Zhong 5 and TAF 46, which were derived from hybrids of wheat with decaploid Thinopyrum ponticum or with hexaploid Th. intermedium. Agrotana was shown to be immune to WSMV and the WCM; the other four partial amphiploids were susceptible to the WCM. Genomic in situ hybridization (GISH) using genomic DNA probes from Th. elongatum (EE, 2n=14), Th. bessarabicum (JJ, 2n=14), Pseudoroegneria strigosa (SS, 2n=14) and T. aestivum (AABBDD, 2n=42) demonstrated that three of the partial amphiploids, Agrotana, OK7211542 and ORRPX, have almost identical alien genome constitutions: all have 16 alien chromosomes, with 8 chromosomes being closely related to the J^s genome and 8 chromosomes belonging to the E or J genomes and no evidence of any S-genome chromosomes. GISH confirmed that the alien genomes of Agrotana and OK7211542, like ORRPX, were all derived from Th. ponticum, and not from Th. intermedium. However, in contrast to Agrotana, ORRPX and OK7211542 were susceptible to the WCM and WSMV. The partial amphiploid Zhong 5 had a reconstituted alien genome composed of 4 S-and 4 J^s-genome chromosomes of Th. intermedium with 6 translocated chromosomes between the S and J^s genomes. This line was highly resistant to WSMV, but was susceptible to the WCM. TAF 46, which contained a synthetic genome consisting of 3 pairs of S-genome chromosomes and 4 pairs of E- or J-genome chromosomes in addition to the 21 pairs of wheat chromosomes, was susceptible to the WCM, but moderately resistant to WSMV. Agrotana offers great potential for transferring WSMV and WCM resistance into wheat. Received: 27 January 1998 / Accepted: 10 February 1998     

Ear photosynthesis, carbon isotope discrimination and the contribution of respiratory CO2 to differences in grain mass in durum wheat

The role of ear photosynthesis in grain filling was studied in a number of durum wheat (Triticum turgidum var durum L.) landraces and varieties from the Middle East, North Africa, and from the collections of ‘Institut National de la Recherche Agronomique’ (INRA, France) and ‘Centro International de Mejora de Maiz y Trigo’ (CIMMYT, Mexico). Plants were grown in the field in a Mediterranean climate. Flag leaves (blade plus sheath) and ears were kept in the dark from 1 week after anthesis to maturity which reduced grain weight by 22.4% and 59.0%, respectively. In a further experiment, the carbon isotope discrimination ratio (Δ) of ear bracts, awns and flag leaves was measured on samples taken at anthesis and on mature kernels. The mean value of Δ for the water soluble fraction of bracts (17.0‰) and awns (17.7‰) were lower than those of leaves (19.5‰) and fairly similar to those of kernels (17.4‰) averaged across all genotypes. Data indicate that most of the photosynthates in the grain come from ear parts and not from flag leaves. In addition, a higher water use efficiency (WUE) of ear parts than of the flag leaf is suggested by their lower Δ values. Gas exchange in ears and flag leaves was measured during grain filling. Averaged over all genotypes, CO₂ diffusive conductance was about five times higher in the flag leaf than in the spike (with distal portions of awns outside the photosynthetic chamber) 2 weeks after anthesis. In absolute terms, the dark respiration rate (R_d) was greater than the net photosynthesis rate (P_n) by a factor of 1.74 in the spike, whereas R_d was much smaller, only 22.1, 65.7 and 24.8% of P_n in blade, sheath and awns, respectively. Data indicate that photosynthesis, and hence the water use efficiency (photosynthesis/transpiration), is greatly underestimated in ears because of the high rates of respiration which diminish the measured rates of net CO₂ exchange. Results of ¹³C discrimination and gas exchange show that genotypes from North Africa have higher WUE than those from the Middle East. The high R_d values of ears as well as their low diffusive conductance suggest that CO₂ from respiration may be used as source of carbon for ear photosynthesis. In the same way, the anatomy of glumes, for example, supports the role of bracts using internal CO₂ as source of photosynthesis. In the first experiment, the Δ in mature grains from culms with darkened ears compared with control culms provided further evidence in support of this hypothesis. Thus, the Δ from kernels of control plants was 0.40 higher than that from ear-darkened plants, probably because of some degree of refixation (recycling) of respired CO₂ in the grains.     

NADH dehydrogenase: a new molecular marker for homoeology group 4 in Triticeae. A map of the 4RS chromosome arm in rye

Summary Structural gene loci encoding the monomeric isozymes nicotin adenin dinucleotide dehydrogenase (NADH dehydrogenase or NDH) have been located on the 4AL, 4B, and 4DS chromosome arms of Triticum aestivum cv Chinese Spring, on the 4RS chromosome arm of Secale cereale cultivars Imperial, King II, Dakold, and Ailes, on the 4S¹ S/7S¹ chromosome of Aegilops longissima, the 4E of Elytrigia elongata, and the CSU-A of Aegilops umbellulata. All the results support the homoeologous relationships among these chromosomes in the five species studied. In addition, a map of the 4RS chromosome arm in cv Ailes has been realized, linking loci Pgm-1 (located on the 4RS chromosome arm) and Ndh-1 (17.91 cM), with an estimated distance between both loci and the centromere of 20.00 cM and 32.12 cM, respectively.     

Analysis of the barley and rice genomes by comparative RFLP linkage mapping

Comparative genetic mapping of rice and barley, both major crop species with extensive genetic resources, offers the possibility of uniting two well-established and characterized genetic systems. In the present study, we screened 229 molecular markers and utilized 110 polymorphic orthologous loci to construct comparative maps of the rice and barley genomes. While extensive chromosomal rearrangements, including inversions and intrachromosomal translocations, differentiate the rice and barley genomes, several syntenous chromosomes are evident. Indeed, several chromosomes and chromosome arms appear to share nearly identical gene content and gene order. Seventeen regions of conserved organization were detected, spanning 287 cM (24%) and 321 cM (31%) of the rice and barley genomes, respectively. The results also indicate that most (72%) of the single-copy sequences in barley are also single copy in rice, suggesting that the large barley genome arose by unequal crossing over and amplification of repetitive DNA sequences and not by the duplication of single-copy sequences. Combining these results with those previously reported for comparative analyses of rice and wheat identified nine putatively syntenous chromosomes among barley, wheat and rice. The high degree of gene-order conservation as detected by comparative mapping has astonishing implications for interpreting genetic information among species and for elucidating chromosome evolution and speciation.     

Rapid identification of Triticeae genotypes from single seeds using the polymerase chain reaction

An easy and quick protocol has been developed for DNA analysis via PCR. Single cereal endosperm or small leaf pieces can be separately processed in several PCR reactions. The resultant PCR patterns are equivalents to those obtained with standard DNA extraction protocols using either specific or random primers. Intra-and inter-specific variability can be detected. This method allows the analysis of a large number of individuals in early stages prior to the plant sowing.     

Phylogenetic analysis of Hordeum marinum Huds. based on nucleotide sequences linked to the vrs1 locus

 More than 100 accessions of Hordeum marinum ssp. marinum, ssp. gussoneanum 2x and ssp. gussoneanum 4x were studied by analyzing the nuclear DNA homologous to the chloroplast translation elongation factor. PCR amplification using the locus-specific primers generated a 932 bp fragment from ssp. marinum, a 915 bp fragment from ssp. gussoneanum 2x, and 915 bp and 931 bp fragments from ssp. gussoneanum 4x. Sequencing of the entire DNAs showed that the 915-bp fragments of diploids and tetraploids are identical, indicating that the diploid probably is the immediate donor of the 915-bp fragment of tetraploids. Although the donor of the 931-bp fragment was not determined, phylogenetic analysis of the data showed that the clade of the 931-bp fragment was included in the H. marinum complex. A MspI site was diagnostic to distinguish ssp. marinum into two clades with disjunct distributions, the Iberian Peninsula (type 2) and other regions (type 1). Nucleotide variation was revealed in the type 2 accessions but all the other clades were highly uniform. Received January 3, 1999 Accepted January 24, 2001     

Allelic diversity of high-molecular-weight glutenin protein subunits in natural populations of Dasypyrum villosum (L.) Candargy

Dasypyrum villosum (L.) Candargy (2n=14, V genome) is a wild, allogamous, diploid grass species that is a potential genetic resource for wheat improvement. The diversity of high-molecular-weight (HMW) glutenin subunits of the seed storage proteins of this species was examined in populations sampled in their natural habitats in Italy and Yugoslavia where the species is widely distributed. The results of selfed progeny tests confirmed that the allelic variation of HMW-glutenin subunits in D. villosum is controlled by a single locus (Glu-V1). Fourteen alleles at Glu-V1 were found among 982 individuals representing 12 populations from Italy and two from Yugoslavia, with a mean of seven alleles per population. Among the 14 Glu-V1 alleles, one produced no HMW-glutenin subunits, ten coded for a single subunit, and three for two subunits. The mobilities of all the subunits in SDS-PAGE gels were greater than that of reference subunit 7 of Triticum aestivum cv Chinese Spring. Eight of the alleles were relatively abundant (mean frequency over all populations ranged from 0.08 to 0.17) and distributed widely among the 14 populations (8 to 14); five of the alleles were rare (0.003 to 0.021) and found in only 1 to 5 populations. The frequencies of two alleles could not be individually estimated because of the similar electrophoretic mobility of their subunits. The multiple-allelic diversity at Glu-V1 was high (H_e ranged from 0.700 to 0.857) but similar from population to population. Overall, about 7% of the total allelic variation was distributed among populations (G_st=0.072), and more than 90% within populations. Whether the allelic variation at Glu-V1 is subject to natural selection is unknown, but the discovery of the homozygous null Glu-V1 alleles in the present study may be useful in pursuing this question. The multiple-allelic diversity in Glu-V1 presents the plant breeder with an opportunity to evaluate and select the most useful alleles for transfer to wheat. The importance of an evaluation genetic diversity in a wild species before interspecific gene transfers are attempted is well illustrated in this study.     

Est-7, a set of genes controlling green tissue esterases in wheat and related species

Summary Analysis using isoelectric focusing of Chinese Spring wheat genetic stocks revealed a set of coleoptile and leaf esterase loci, designated Est-7, on the long arms of the group 2 chromosomes. A survey of 38 other hexaploid genotypes revealed onyl a single variant, at Est-D7. Homoeoloci were found on chromosome (arm) 2HL of Hordeum vulgare, 2RL of Secale cereale, 2R ^m of S. montanum, 2U of Aegilops umbellulata, 2E of Agropyron elongation and 2V of Dasypyrum villosa.