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Nuclear genes control changes in the organization of the mitochondrial genome in tissue cultures derived from immature embryos of wheat.

Although the mitochondrial genomes of the Chinese Spring and Aquila varieties of wheat are normally similar in organization, this is not so in tissue cultures initiated from their immature embryos where the mitochondrial genomes of both are rearranged and in different, characteristic, ways. However, the mitochondrial genomes of tissue cultures of reciprocal F1 crosses between these varieties were almost identical to one another, showing that nuclear genes control the rearrangement processes. These rearrangements are either due to the appearance of new structures or else result from changes in the relative amounts of subgenomic components. The severe reduction in the amount of certain molecular configurations in tissue cultures from reciprocal crosses is probably due to the presence of dominant information in the Aquila nuclear genome. Data obtained from tissue cultures initiated from F2 embryos of the cross Aquila x Chinese Spring suggest that at least two complementary genes are involved in this control. In contrast, the presence of new molecular arrangements appears to be under the control of a dominant allelic form of a Chinese Spring gene or genes. Thus, this study demonstrates that at least two sets of nuclear genes control the reorganization of the mitochondrial genome which occurs when tissue cultures are initiated from the immature embryos of wheat.

Blotting, Southern

Low quality roughages for steers grazing wheat pasture. I. Effect on weight gains and bloat.

The effect of feeding low quality roughages (LQR) on live and carcass weight gains and the incidence and severity of bloat of stocker cattle grazed on wheat pasture was evaluated in a 3 yr study. One hundred eighty-five steer calves (172 kg mean initial weight) grazed clean-tilled wheat pasture and were either fed no LQR or had ad libitum access to wheat straw (WS) or sorghum-Sudan hay (SS). Grazing periods were (I) fall grazing, (II) winter grazing, (III) period of lush spring growth of wheat forage and (IV) period of advancing forage maturity and declining quality. Mean dry matter (DM), crude protein and acid detergent fiber (ADF) content (percentage of DM) of wheat forage averaged across years ranged, respectively, from 23.8 to 33.0, 19.8 to 26.4 and 21.5 to 27.7. Mean daily consumption (kg DM/head) of WS and SS by steers ranged from .076 to .100 and .199 to .248, respectively. Live and carcass weight gains of steers during Periods I through III (i.e., the usual wheat pasture grazing period) were not influenced (P greater than .05) by treatments. Carcass weight gains were about 74% of live weight gains. Bloat was observed only during the last 2 wk of Period III of the first year. The incidence (steer days of bloat) and severity (bloat score) of control, WS- and SS-fed steers were 9.5 and 1.2, .5 and .5 and 2.0 and 1.0, and were not different (P greater than .05) among treatments. Intake of WS and SS [g/body weight (BW).75kg] during Periods I to III was, respectively, only about 5 and 12% of roughage intakes (i.e., 37.5 g/BW.75kg) reported in the literature to "effectively control" or aid the prevention of bloat. It seems unlikely that LQR consumed to amounts similar to those of this study would control bloat of stocker cattle on wheat pasture.

Animals

Linked sucrose synthase genes in group-7 chromosomes in hexaploid wheat (Triticum aestivum L.).

A cDNA library from developing wheat endosperm was screened for sucrose-synthase clones using a maize cDNA probe corresponding to the Sh1 locus under non-stringent conditions. Five positive clones were isolated and initially classified into two types on the basis of their relative ability to hybridize with the probe and of their partial restriction maps. Determination of the nucleotide sequences indicated homology between the two types of wheat clones, with type 1 showing higher homology to the maize Sh1 locus than to type-2 sequences. The inserts cloned in plasmids pST8 (type 1) and pST3 (type 2) were used as probes to determine the chromosomal locations of the two types of genes. DNAs from compensated nulli-tetrasomic and ditelosomic lines of wheat cultivar Chinese Spring were cleaved with EcoRI and analysed in Southern blots. DNA segments of the two types were thus identified in the short arms of chromosomes 7A, 7D, and, possibly, 7B. The two types of linked loci have been designated Ss1 and Ss2, respectively.

Amino Acid Sequence

[Nutrient composition of some newly bred high protein and/or high lysine grains and their digestibility determined on growing pigs].

With the view to complementing the feedstuff data store and, consequently, to further improving the GDR Feed Evaluation System, some newly bred high-protein and/or high-lysine cereal varieties and strains (spring barley, winter wheat, maize) were studied for their nutrient composition and digestibility. Apart from from WEENDER's feed analysis technique, more recent methods were applied to determine total fat (after HCl treatment), carbohydrates (enzymatic method), lignin and amino acids. The digestibility of the nutrients was determined using growing pigs of different live weight, the test rations being made up of the cereals under and supplementations of limiting amino acids as well as vitamins and minerals. In comparison with the values from currently applied tables, the newly bred strains and varieties proved to have markedly higher contents of crude protein, digestible crude protein, lysine and energetic feed equivalents. Compared to crude fat, the total fat values proved markedly higher in the barley and wheat samples. The readily soluble and easily hydrolizable carbohydrates found with the new analysis procedure suggested, were 100% digestible in all cereal samples used. Lignin proved the constituent most difficult to digest and must be regarded as virtually undigestible in the case of pigs.

Animal Nutritional Physiological Phenomena

Detection of maize DNA sequences amplified in wheat.

Genomic in situ hybridization to somatic metaphase chromosomes of hexaploid wheat cv. Chinese Spring using biotinylated maize genomic DNA as a probe revealed the existence of amplified maize DNA sequences in five pairs of chromosomes. The in situ hybridization sites were located on chromosomes 1A, 7A, 2B, 3B, and 7B. One pair of in situ hybridization sites was also observed in hexaploid oat. The locations and sizes of in situ hybridization sites varied among progenitor species.

Chromosome Mapping

Genetics and evolution of multilocus isozymes in hexaploid wheat.

Aneuploid genetic studies of isozyme variation in cv Chinese Spring have disclosed that numerous enzymes of hexaploid wheat exist in multiple molecular forms as a direct consequence of polyploidy. Sixty-nine isozyme structural genes have been identified to date. Two of these belong to a duplicate set and at least 54 to triplicate sets of paralogous genes that are located one each in related chromosomes in different genomes. Each of these gene sets encodes either two or three isozymes. The role of regional gene duplication in the production of multilocus isozymes in hexaploid wheat is as yet poorly understood, although a considerable amount of indirect evidence suggests that a large number of isozymes are encoded by genes that were produced by ancient regional gene duplication events in a genome ancestral to the genomes now present in the species. A full assessment of the role of regional gene duplication in the production of hexaploid wheat isozymes must await further studies. The isozyme structural gene locations thus far determined indicate that the gene synteny relationships that existed in the ancestral wheat genome are in large part conserved in each of the three genomes of cv Chinese Spring and that the genetic content of most individual chromosome arms has also been in large part conserved.

Biological Evolution

Fluorescent in situ hybridization and C-banding analyses of highly repetitive DNA sequences in the heterochromatin of rye (Secale montanum Guss.) and wheat incorporating S. montanum chromosome segments.

The molecular characterization of C-banded regions of Secale montanum Guss. by means of in situ hybridization was performed in order to provide new information about their chromosome structure relative to cultivated rye, Secale cereale L. Accurate identification of individual chromosomes was achieved using simultaneous and (or) successive fluorescent in situ hybridization (FISH) and C-banding. FISH identification was performed using total rye DNA, three highly repetitive rye DNA sequences (pSc119.2, pSc74, and pSc34), and the ribosomal RNA probes pTa71 (18S, 5.8S, and 26S rDNA) and pTa794 (5S rDNA). FISH was also used to identify the chromosome segment involved in two spontaneous translocation lines recovered from a 'Chinese Spring'--S. montanum wheat-rye addition line. FISH analysis revealed the exact translocation breakpoints and allowed the identification of the transferred rye segments. The value of this type of analysis is discussed.

Chromosome Banding

[Effect of plant residues on the parasitic activity of soilborne pathogens and the saprophytic microflora of the soil. III. Influence of rye and rape grown as winter catch crop on the incidence of Cercosporella herpotrichoides Fron (author's transl)].

In a five-years field trial, the influence of two winter catchcrops, rye and rape, upon the incidence of Cercosporella herpotrichoides has been studied. The winter catch-crops had been repeated three times, followed either by stubble-crop or without in fall of the fourth year. The variants (blocks) had been completed by two variants without catch-crop, either carefully cultivated or overgrown with weeds. Each of the four blocks had been divided three times to be grown with three different crop sequences containing 80%, 60% (with the insusceptible corn), or 40% haulm fruit, respectively. Each of the 12 variants had been divided into two parts by continuous fallow stripe, one part being artificially infested with the pathogen in the beginning (1st winter wheat). In the 2nd, 3rd, and 4th year, only spring crops were grown, followed by winter wheat in the 5th year. The final exploration resulted in a medium to heavy degree of infection, being significantly higher in the 80% cereal crop sequence than in the 60% and 40% sequences. Concerning the catch-crops, infection in the rye variants was equal or significantly lower than in the carefully cultivated variants without catch-crop. On the other hand, infection of the rape variants had become significantly higher than that of the catch-crop-free ones, about as heavily as in the weed-infested variants. From fall 1966 to summer 1969, the development of the saprophytic soil microflora and several other edaphic parameters had been investigated. Remarkable differences in intensity of disintegration have been observed under the influence of the main crops, cereals or potatoes, respectively, but no differences could be stated resulting from the matter of disintegration--residues of rye or rape--, which might interpret the contrasting effects on the pathogen. Too, no indication for antagonistic activity of bacteria or actinomycetes has been noted. The results are compared with known facts of Cercosporella and of other soil borne pathogens. The difference in biology of Cercosporella on the one hand, and of root infecting fungi on the other hand seems to be most important to explain the results obtained.

Actinomycetales

The characterization of cDNA clones coding for wheat storage proteins.

Poly(A)+ RNA isolated from the developing wheat endosperm var. Chinese Spring, has been used as template for the construction of a cDNA library. Within the library, clones have been identified by in vitro translation of hybrid-selected mRNA which encode alpha/beta gliadin related sequences and gamma-gliadin related sequences. The DNA sequence of one such clone has been determined and it shows homology with that of a clone encoding a barley storage protein, B-hordein. The sequence includes a tandem DNA repeat which is discussed in relation to the generation of diversity within the gliadins.

Amino Acid Sequence

The developmental stage of inactivation of rye origin rRNA genes in the embryo and endosperm of wheat x rye F1 hybrids.

To identify the developmental stage during which the preferential inactivation of rRNA genes from the rye parent occurs in wheat x rye hybrids, nucleolar activity was evaluated in the embryo and endosperm of developing seeds of the hybrids. The hybrids were obtained from crosses of euploid and aneuploid lines of hexaploid wheat, Triticum aestivum cv. Chinese Spring, with rye, Secale cereale cv. Centeio do Alto. The number of nucleolar organizing regions (NORs) and nucleoli present in the embryo and endosperm cells of wheat, and wheat x rye F1 hybrids, at different times after fertilization was scored by silver staining. The inactivation of rDNA of rye origin in F1 hybrids occurs simultaneously in the embryo and in the endosperm between 4 and 5 days after fertilization, when these have been through six and 10 cell cycles respectively. We conclude that the genomic interactions leading to the inactivation of the rye origin rDNA is a time-dependent process, related to the developmental stage and independent of the number of cell cycles (DNA replication rounds) they have been through.

Cell Count

Identification of wheat (Triticum aestivum L.) chromosomes with genes controlling the level of nitrate reductase, nitrite reductase, and acid proteinase using the Chinese Spring-Hope substitution lines.

The levels of nitrate reductase, nitrite reductase, and acid proteinase were compared in the primary leaves of 8-day-old wheat seedlings of Chinese Spring, Hope, and the 21 disomic substitution lines of Hope in Chinese Spring. Two chromosomes, 7B and 7D, were considered to contain genes controlling the level of nitrate reductase. Substitution of Hope chromosome 7B caused a highly significant increase in the in vitro stability of nitrate reductase. Nitrite reductase appeared to be controlled by two major genes, located on chromosomes 4D and 7D, and two minor genes, located on chromosomes 3D and 5A. In the case of acid proteinase, substitution of chromosome 1D caused a significant reduction in enzyme activity.

Chromosome Mapping

Genetic control of the mitochondrial form of superoxide dismutase in hexaploid wheat.

Extracts of mature grains of a large number of aneuploid derivatives of Triticum aestivum cv. Chinese Spring and of the members of five wheat-alien chromosome addition series were subjected to isoelectric focusing in polyacrylamide gels in order to study the genetic control of superoxide dismutase (SOD). Evidence was obtained that homologous structural genes for the mitochondrial form of SOD are located in the long arms of the homologous group 2 chromosomes of Chinese Spring and in chromosome 2R of Secale cereale cv. Imperial. The SOD gene loci located in chromosomes 2A, 2B, 2D, and 2R were designated Sod-A1, Sod-B1, Sod-D1, and Sod-R1, respectively. Chromosome-arm pairing data indicate that 2DL is not homologous to either 2AS or 2BL. The results of this study suggest, however, that 2BL is partially homologous to both 2AL and 2DL.

Aneuploidy

Chromosome painting in plants: in situ hybridization with a DNA probe from a specific microdissected chromosome arm of common wheat.

We report here on the successful painting of a specific plant chromosome within its own genome. Isochromosomes for the long arm of chromosome 5 of the wheat B genome (5BL) were microdissected from first meiotic metaphase spreads of a monoisosomic 5BL line of the common wheat Triticum aestivum cv. Chinese Spring. The dissected isochromosomes were amplified by degenerate oligonucleotide-primed PCR in a single tube reaction. The amplified DNA was used as a complex probe mixture for fluorescent in situ hybridization on first meiotic metaphase spreads of lines carrying 5BL as a distinctive marker. Hybridization signals were observed, specifically, along the entire 5BL. In some of the cells, labeling was also detected in two bivalents, presumably those of the 5B "homoeologues" (partial homologues) found in common wheat (5A and 5D). The probe also revealed discrete domains in tapetal nuclei at interphase, further supporting the probe's high specificity. These data suggest that chromosome and homoeologous group-specific sequences are more abundant in 5BL than genome-specific sequences. Chromosome-painting probes, such as the one described here for 5BL, can facilitate the study of chromosome evolution in polyploid wheat.

Base Sequence

Effects of corn oil and wheat brans on bile acid metabolism in rats.

High concentrations of colonic bile acids may promote tumor formation. Some studies have found that high levels of dietary fat increase fecal bile acid excretion, whereas others report no effect. Wheat bran appears to reduce fecal bile acid concentration. This study was conducted to determine the effect of different dietary fat levels and types of wheat bran on bile acid metabolism. Rats were fed diets containing either no fiber, 2% cholestyramine (CHO) or brans of hard red spring, soft white winter or durum wheat--at both a 5 or 20% fat level. Animals were fed for 7 wk, and feces were collected in the last week. Wheat bran (all types) significantly increased fecal mass approximately fourfold, and CHO significantly increased fecal mass twofold compared to the fiber-free diet. Increasing the fat level did not increase fecal bile acid excretion, nor did the addition of wheat bran. Addition of CHO, however, more than doubled it. CHO increased fecal bile acid concentration, all wheat brans decreased it and fat level had no effect. Bile acid pool size was increased slightly by fat level and cholestyramine feeding but not by wheat brans. These results indicate that fat level slightly alters bile acid metabolism but that wheat brans do not.

Animals

High-resolution cytological mapping of the long arm of chromosome 5A in common wheat using a series of deletion lines induced by gametocidal (Gc) genes of Aegilops speltoides.

Gametocidal (Gc) genes of Aegilops in the background of the wheat genome lead to breakage of wheat chromosomes. The Q gene of wheat was used as a marker to select 19 deletion lines for the long arm of chromosome 5A of common wheat, Triticum aestivum cv. Chinese Spring (CS). The extents of deleted segments were cytologically estimated by the C-banding technique. The DNAs of deletion lines were hybridized with 22 DNA probes recognizing sites on the long arm of the chromosome (5AL) to determine their physical order. Based on the breeding behavior of the deletion lines, the location of a novel gene (Pv, pollen viability) affecting the viability of the male gamete was deduced. The segment translocated from 4AL to 5AL in CS was cytologically estimated to represent 13% of the total length of 5AL. Although DNA markers were almost randomly distributed along the chromosome arm, DNA markers located around the centromere and C-banded regions were obtained only rarely. Some deletion lines were highly rearranged in chromosome structure due to the effect(s) of the Gc gene. Applications of Gc genes for manipulating wheat chromosomes are discussed.

Blotting, Southern

Genetic effect of the Ph1 locus on transcriptome atlas of anther development-related genes, meiotic chromosome behavior and agronomic traits in bread wheat.

Proper spatiotemporal expression of meiosis-related genes (MRGs) and other male-microsporogenesis/microgametogenesis-related genes (MMRGs) is crucial for normal anther development, yet their expression patterns remain largely unknown in wheat. The Ph1 locus in wheat is known to contain the Ph1 gene that plays a dual role in promoting pairing between homologous chromosomes but repressing pairing between homoeologous chromosomes, but its genetic function is still unclear. Here, we investigated these issues by conducting a comprehensive transcriptome analysis during wheat anther development in Chinese Spring (CS) and its ph1b deletion mutant under greenhouse and field conditions. Our results revealed that MRGs and MMRGs are predominantly expressed during pre-meiosis stages, with MMRGs also being highly expressed in meiotic-II. Gene co-expression analysis showed that C2H2 and B3 transcriptional factors (TFs) are associated with MRGs, and MYB regulators interacted mainly with MMRGs during microgametogenesis. Deletion of genes within the Ph1 locus failed to induce compensatory transcriptional activation of their homoeologous counterparts, while genes outside the Ph1 locus showed environmental-specific responses, especially during meiotic-II and mature pollen stages. Notably, early disjunction of bivalent chromosomes is a primary factor leading to defective meiocytes during metaphase I. Furthermore, the ph1b deletion mutant exhibited a substantially delayed heading date, potentially contributing to environment-stable and environment-specific alterations in fertility and grain-related traits. Our study highlights the significant impact of the Ph1 locus on the transcriptome during anther development, and a previously unheeded effect on meiotic chromosome pairing and agronomic traits, suggesting potential for genetic manipulations within the Ph1 locus for wheat improvement.

Triticum

Chromosomal locations of the genes for histones and a histone gene-binding protein family HBP-1 in common wheat.

The chromosomal locations of the genes in common wheat that encode the five histones and five members of the HBP (histone gene-binding protein)-1 family were determined by hybridizing their cloned DNAs to genomic DNAs of nullitetrasomic and telosomic lines of common wheat, Triticum aestivum cv. Chinese Spring. The H1 and H2a genes are located on different sets of homoeologous chromosomes or chromosome arms, namely, 5A, 5B and 5D, and 2AS, 2BS and 2DS, respectively. Genes for the other histones, H2b, H3 and H4, are found in high copy number and are dispersed among a large number of chromosomes. The genes for all members of the HBP-1 family are present in small copy numbers. Those for HBP-1a(1) are located on six chromosome arms, 3BL, 5AL, 5DL, 6AL, 6BS and 7DL, whereas those for each HBP-1a(c14), 1a(17), 1b(c1), and 1b(c38) are on a single set of homoeologous chromosome arms; 4AS, 4BL, 4DL; 6AS, 6BS, 6DS; 3AL, 3BL, 3DL; and 3AS, 3BS, 3DS, respectively. The genes for histones H1 and H2a, and for all members of the HBP-1 family except HBP-1a(1) are assumed to have different phylogenetic origins. The genes for histone 2a and HBP-1a(17) are located in the RFLP maps of chromosomes 2B and 6A, respectively. Gene symbols are proposed for all genes whose chromosomal locations have been determined.

Aneuploidy