The role of awns in the resistance of cereals to the grain aphid, Sitobion avenae

Six genotypes of awned spring wheat were more resistant to Sitobion avenae than de-awned wheat. Aphids that fed on awns were up to 22% less fecund and were more likely to be dislodged than aphids that fed elsewhere on an ear. These two factors reduced aphid population growth on awned earing plants in the field to a third of that observed on awnless plants. As awns may also benefit yield and are easy to select for it is recommended that awned winter wheats could be bred and used to reduce the incidence of outbreaks of S. avenae.     

The DROOPING LEAF and OsETTIN2 genes promote awn development in rice

The awn is a long needle‐like appendage that, in some grass species, is formed on the lemma that encloses floral organs together with the palea. In rice, most wild species and most strains of Oryza sativa ssp. indica generate an awn, whereas most strains of O. sativa ssp. japonica do not. In japonica, the long‐awn characteristic appears to have been lost during domestication and breeding programs. Here, we found that the genes DROOPING LEAF (DL) and OsETTIN2 (OsETT2) are involved in awn development in the awned indica strain Kasalath. Genetic analyses and RNA‐silencing experiments indicate that DL and OsETT2 act independently in awn formation, and that either gene alone is not sufficient for awn development. Scanning electron microscopy revealed that the top region of the lemma (a putative awn primordium) is larger in an awned floret than in an awnless floret. OsETT2 is expressed in the awn primordium in the awned indica floret, but not in the awnless japonica floret except in the provascular bundle. DL is expressed underneath the primordium at similar levels in both indica and japonica florets, suggesting non‐cell‐autonomous action. We hypothesize that loss of expression of OsETT2 in the awn primordium is probably associated with the failure of awn formation in japonica strains.     

Ear and flag leaf photosynthesis of awned and awnless Triticum species

The net photosynthetic rates of ears and flag leaves of awned and awnless wheats were measured by infra-red gas analysis in a controlled environment experiment. At 107 W m^-2 the rates of net photosynthesis of the ears of two awned Triticum aestivum lines were two to three times greater than those of their isogenic awnless counterparts. The net rates of photosynthesis of the flag leaves of all four lines were, however, similar. Net photosynthetic rates of ears of T. durum and T. turgidum, which have much larger awns, were considerably higher than those of T. aestivum. Between 70 and 90% of carbon-14 assimilated by the flag leaves and by the ears of T. aestivum was translocated to the grains, with no effect of awns in this respect. In a glasshouse experiment with detached shoots the contributions of the component organs to gross photosynthesis were determined by carbon-14 labelling. Ears of awnless lines of T. aestivum contributed about 10% to the photosynthesis of the organs above the node of the penultimate leaf and awns increased this to about 18%. Fixation by awned ears of T. durum and T. turgidum was 21–29% and of this 78–86% was contributed by the awns. In both experiments the contribution of the flag leaf to shoot photosynthesis was greater in T. aestivum than in the other species.     

Photosynthetic and translocation pattern in contrasting winter wheat varieties

The rates of photosynthesis and patterns of translocation of ¹⁴C from the flag leaves of tall and semi-dwarf wheat varieties, with and without awns, were compared in a field experiment. The rates of photosynthesis of the awnless varieties fell during the 14 days after anthesis but those of the awned varieties did not change. Wide differences in translocation pattern of ¹⁴C supplied at anthesis were also found between varieties, but the differences in distribution pattern of ¹⁴C supplied 14 days later were less. At anthesis, the high yielding semi-dwarf selection translocated most carbohydrate to the ears and awned selections least. It is suggested that varietal differences in translocation pattern are determined by differences in the capacity of the grain to store carbohydrate.     

Genetic mapping reveals a dominant awn-inhibiting gene related to differentiation of the variety anathera in the wild diploid wheat <Emphasis Type="Italic">Aegilops tauschii</Emphasis>

Aegilops tauschii, a wild wheat relative, is the D-genome donor of common wheat. Subspecies and varieties of Ae. tauschii are traditionally classified based on differences in their inflorescence architecture. However, the genetic information for their diversification has been quite limited in the wild wheat relatives. The variety anathera has no awn on the lemma, but the genetic basis for this diagnostic character is unknown. Wide variations in awn length traits at the top and middle spikes were found in the Ae. tauschii core collection, and the awn length at the middle spike was significantly smaller in the eastward-dispersed sublineage than in those in other sublineages. To clarify loci controlling the awnless phenotype of var. anathera, we measured awn length of an intervariety F₂ mapping population, and found that the F₂ individuals could be divided into two groups mainly based on the awn length at the middle of spike, namely short and long awn groups, significantly fitting a 3:1 segregation ratio, which indicated that a single locus controls the awnless phenotype. The awnless locus, Anathera (Antr), was assigned to the distal region of the short arm of chromosome 5D. Quantitative trait locus analysis using the awn length data of each F₂ individual showed that only one major locus was at the same chromosomal position as Antr. These results suggest that a single dominant allele determines the awnless diagnostic character in the variety anathera. The Antr dominant allele is a novel gene inhibiting awn elongation in wheat and its relatives.     

The Effect of Substitution of Chromosome 5A with an Extra Chromosome 5B in the Progeny of a Somaclonal Common Wheat Line Monosomic for 5A

Cytogenetic analysis was employed in studying the cause of generation of fertile awned forms in the progeny of plants, which were selected from a speltoid somaclonal wheat line monosomic for chromosome 5A (2n = 41 = 20II + I), had speltoid spikes, and were reproduced by self pollination. On cytogenetic and genetic evidence, chromosome 5A was eliminated and the copy number of chromosome 5B increased in the plants examined. The appearance of an extra chromosome 5B is probably caused by nondisjunction of bivalent 5B in the presence of a telocentric originating from the long arm of chromosome 5A. A difference in meiotic segregation was observed for univalent chromosomes 5A and 5B.     

Effect of awns and drought on the supply of photosynthate and its distribution within wheat ears

The presence of awns doubled the net photosynthetic rate of wheat ears and also increased the proportion of ¹⁴CO₂ assimilated by the ear that moved to the grain. The effect of water supply on photosynthesis and movement of assimilates was greater for leaves than ears, so that drought increased the proportion of assimilate contributed by ear photosynthesis to grain filling from 13% to 24% in the awnless ears, and from 34% to 43% in the awned ears. ¹⁴C assimilated by the ears was most important to the economy of the upper spikelets and to the distal florets in each spikelet, whereas flag leaf assimilate went mainly to the spikelets in the lower half of the ear, and to the proximal florets. Awns increased grain yield in the dry but not in the irrigated treatment, despite the large contribution of awned ears to grain filling. Either the supply of assimilate did not limit grain yield when water supply was not limiting, or there were compensating disadvantages to awns. However, they did not seem to have any adverse effect on the development of the upper florets, nor did they reduce grain number per ear.     

Stable wheat transformation obtained without selectable markers

Transgenic wheat plants without the selectable marker gene were obtained either in the presence or in the absence of selective pressure during the transformation protocol. When using hygromycin as selective agent in a co-transformation experiment involving a mixture of plasmids pGL2, containing the hpt gene, and pAI₁Gus, containing the uidA gene, 3 out of 19 transgenic wheat plants had the uidA gene alone as shown by Southern blots. The gene was transmitted to the progeny following Mendelian rules. Segregation and loss of the selectable marker gene was also found in three out of six events from other experiments where high-molecular-weight glutenin genes were expressed or over-expressed. On the other hand, in 7 experiments where no selective pressure was applied and that involved 1016 bombarded explants, 23 transgenic wheat plants were obtained. The uidA gene was stably integrated as suggested by its transmission to the progeny.these authors contributed equally to the work     

A cytological study of plants growing under exposure to different radiation levels

Genetically significant consequences of emergency at the Chernobyl Atomic Power Plant have been studied on wheat and rye plants. Plants grown in the 30-km zone of the plant after its emergency are determined to have high frequency of chromosome aberrations reaching 2.666% in rye and 1.075-2.572% in wheat and 2.235-3.187% as dependent on the variety, biotype (awnless, semi-awned and awned) and places of occurrence. These levels of aberrations are 5.34, 2.42-5.78 and 4.07-6.71 times higher than their frequency in control plants (0.499, 0.444 and 0.475%, respectively).     

Genetic analysis of the traits introgressed from <Emphasis Type="Italic">Aegilops speltoides</Emphasis> Tausch. to bread wheat and determined by chromosome 5A genes

A winter bread wheat accession from the Arsenal collection was genetically examined to study the results of introgression, which substantially changed the physiological and morphological traits of the original spring cultivar Rodina. Apart from its winter habit, the accession was characterized by awned speltoid spikes, suggesting introgression into chromosome 5A, which carries marker genes in the order Vrn-A1-Q-B1. Genetic analysis showed that the chromosome fragment introgressed from Aegilops speltoides recombined well with the homeologous region of bread wheat chromosome 5A in the region between the Vrn-A1 and Q genes. Recombination between the Vrn-A1 and B1 genes was not detected, and it was assumed that the order of the marker genes of chromosome 5A was inverted to produce Q-Vrn-A1-B1. When the winter introgression line was crossed with Triticum spelta L., an interaction of two dominant genes determining the spike character was for the first time detected in F₁, increasing the spike length and the number of spikelets, and followed with transgression in F₂. It was assumed that Ae. speltoides had a homeoallelic speltoid gene, which was designated as Q ^S .     

Molecular mapping of hybrid necrosis genes Ne1 and Ne2 in hexaploid wheat using microsatellite markers

Hybrid necrosis is the gradual premature death of leaves or plants in certain F₁ hybrids of wheat (Triticum aestivum L.), and it is caused by the interaction of two dominant complementary genes Ne1 and Ne2 located on chromosome arms 5BL and 2BS, respectively. To date, molecular markers linked to these genes have not been identified and linkage relationships of the two genes with other important genes in wheat have not been established. We observed that the F₁ hybrids from the crosses between the bread wheat variety ‘Alsen’ and four synthetic hexaploid wheat (SHW) lines (TA4152-19, TA4152-37, TA4152-44, and TA4152-60) developed at the International Maize and Wheat Improvement Center (CIMMYT) exhibited hybrid necrosis. This study was conducted to determine the genotypes of TA4152-60 and Alsen at the Ne1 and Ne2 loci, and to map the genes using microsatellite markers in backcross populations. Genetic analysis indicated that Alsen has the genotype ne1ne1Ne2Ne2 whereas the SHW lines have Ne1Ne1ne2ne2. The microsatellite marker Xbarc74 was linked to Ne1 at a genetic distance of 2.0 cM on chromosome arm 5BL, and Xbarc55 was 3.2 cM from Ne2 on 2BS. Comparison of the genetic maps with the chromosome deletion-based physical maps indicated that Ne1 lies in the proximal half of 5BL, whereas Ne2 is in the distal half of 2BS. Genetic linkage analysis showed that Ne1 was about 35 cM proximal to Tsn1, a locus conferring sensitivity to the host selective toxin Ptr ToxA produced by the tan spot fungus. The closely linked microsatellite markers identified in this study can be used to genotype parental lines for Ne1 and Ne2 or to eliminate the two hybrid necrosis genes using marker-assisted selection. Mention of trade names or commercial products in this article is solely for the purpose of providing specific information and does not imply recommendation or endorsement by the US Department of Agriculture.     

Effect of genome, induction medium and temperature pretreatment on green plant production in durum ( Triticum turgidum var. durum ) x bread wheat ( Triticum aestivum L. em Thell) crosses

The recalcitrancy of durum wheat (Triticum turgidum var. durum) to anther culture, was attempted to be overcome by transferring the responsible genes form bread wheat B-genome to the respective on durum wheat, determining an appropriate induction medium and clarifying the necessity of cold pretreatment. For this, three durum wheat cultivars were crossed to two bread wheat (Triticum aestivum L. em Thell) cultivars. The resulting F₁ plants and their original cultivars were grown in the field and anthers at the appropriate microspore stage were cultured on potato-2 and W₁₄ media with and without low temperature pretreatment. No green plants were produced from the parental durum wheat cultivars. In contrast, green plants were produced from the F₁ plants. The best results in three of the four F₁ hybrids were recorded when potato-2 was used as induction medium. A more variable response of the examined genotypes was noticed with respect to temperature pretreatment. Regarding green plant production, a negative effect of cold pretreatment was observed in two of the F₁ hybrids when they were cultured on potato-2. Chromosome counts on root tips from the resulting green plants revealed that they all carried D-genome chromosomes. The last observation could suggest that D-genome chromosomes are necessary for anther culture response in wheat. Yet, the production of one green plant with 15 chromosomes may indicate that the development of extracted durum genotypes from bread wheat genotypes with good response to in vitro anther culture might be possible. Further work however, is needed for this to be verified.     

Construction of Immune Lines with Complex Resistance to Leaf Rust and Powdery Mildew in Common Spring Wheat Cultivar Saratovskaya 29

Immune lines resistant both to leaf rust and to powdery mildew were constructed on the basis of common wheat cultivar Saratovskaya 29. Synthetic wheat Triticum timopheevii/Aegilops squarrosa (AAGGDD, 2n = 42) of Savov (Bulgaria) was used as a source of resistance genes. Using cytological analysis of BC₂, we selected resistant plants (21") free from meiosis 1 (M1) defects. With these plants and continuous selection, BC₈–BC₉ immune lines were obtained. The lines were shown to carry new resistance genes differing from the known ones, and were proposed as donors of immunity to the diseases.     

Antifungal Activity of Aqueous Extracts of the Leaves of Cowparsnip and Comfrey

We found that extracts from the leaves of medicinal comfrey and cowparsnip strongly inhibit the germination of Erysiphe graminisconidia and uredospores of Puccinia graminis. Spraying wheat seedlings with these extracts, in contrast to the irrigation of soil, markedly diminished infection in plants with powdery mildew. Antifungal activity in vitroand protective activity (when plants were sprayed) correlated with the level of phenolic compounds in these extracts. Experiments with healthy plants have demonstrated that the photosynthetic apparatus of wheat plants is stimulated by extracts. Spraying seedlings with the extracts resulted in an increased rate of O₂evolution calculated per unit of chlorophyll, an increase in the ratio (F_M– F_T)/F_Tin the experiments that recorded slow fluorescence induction, an increase in the relative light intensity of band A, and a decrease of relative intensity of band Cin experiments with thermoluminescence of wheat leaves. These results provide evidence that the protective activity of comfrey and cowparsnip extracts is associated with their action on the pathogenic fungus and with the activation of natural defense reactions of the host plant.     

Variation and geographical distribution of the genotypes controlling the diagnostic spike morphology of two varieties of Aegilops caudata l

Aegilops caudata L. is an annual wild relative of wheat distributed over the northeastern Mediterranean basin. It consists of two taxonomic varieties, var. typica with awnless lateral spikelets and var.polyathera with awned lateral spikelets. To clarify the variation and the geographical distribution of the genotypes controlling the diagnostic spike morphology of the two taxonomic varieties, three crossing experiments were carried out. First, two varieties collected from nine sympatric populations in the Aegean islands were crossed reciprocally. All of the F1 hybrids were var. typica and the segregation ratio in the F2 generation was 3 typica: 1 polyathera. Secondly, 13 typica accessions collected from the entire distribution area of the variety were crossed with a common polyathera accession. The F1 hybrids involving eight typica accessions from Greece and West Anatolia were var. typica, while those involving five typica accessions from East Anatolia, Syria and Iraq were var. polyathera. Thirdly, the typica F1 hybrids between the Aegean and the Syrian typica accessions were backcrossed to the latter. Four of the seven BC1F1 plants obtained were var. typica, but the other three were var. polyathera. Based on these results, the following two conclusions were made. First, the awnless lateral spikelets characteristic of var. typica are due to two different genotypes: one is a dominant allele suppressing awn development on lateral spikelets and the other is a recessive allele(s) for awnless lateral spikelets with no dominant suppressor allele. Secondly, the former genotype occurs only in the western region of the distribution area of the species, while the latter occurs in the eastern region. The present results and the recent palaeopalynological evidence also suggested that var. polyathera, with more awns than var. typica, rapidly colonized Central Anatolia from the Levant or East Taurus/Zagros mountains arc after the last glacial period.     

Generation of marker‐free transgenic hexaploid wheat via an Agrobacterium‐mediated co‐transformation strategy in commercial Chinese wheat varieties

Genotype specificity is a big problem lagging the development of efficient hexaploid wheat transformation system. Increasingly, the biosecurity of genetically modified organisms is garnering public attention, so the generation of marker‐free transgenic plants is very important to the eventual potential commercial release of transgenic wheat. In this study, 15 commercial Chinese hexaploid wheat varieties were successfully transformed via an Agrobacterium‐mediated method, with efficiency of up to 37.7%, as confirmed by the use of Quickstix strips, histochemical staining, PCR analysis and Southern blotting. Of particular interest, marker‐free transgenic wheat plants from various commercial Chinese varieties and their F₁ hybrids were successfully obtained for the first time, with a frequency of 4.3%, using a plasmid harbouring two independent T‐DNA regions. The average co‐integration frequency of the gus and the bar genes located on the two independent T‐DNA regions was 49.0% in T₀ plants. We further found that the efficiency of generating marker‐free plants was related to the number of bar gene copies integrated in the genome. Marker‐free transgenic wheat plants were identified in the progeny of three transgenic lines that had only one or two bar gene copies. Moreover, silencing of the bar gene was detected in 30.7% of T₁ positive plants, but the gus gene was never found to be silenced in T₁ plants. Bisulphite genomic sequencing suggested that DNA methylation in the 35S promoter of the bar gene regulatory region might be the main reason for bar gene silencing in the transgenic plants.     

Inheritance of hypersensitive resistance to Bean yellow mosaic virus in narrow-leafed lupin (Lupinus angustifolius)

In narrow‐leafed lupin (Lupinus angustifolius), segregation for the necrotic (systemic hypersensitive) response to infection with a necrotic strain of Bean yellow mosaic virus (BYMV‐N) was studied in progeny plants from six crosses. The parents were two cultivars that always developed necrosis when infected (Danja and Merrit) and two genotypes that always responded without necrosis (90L423‐07‐13 and P26697). In the four possible combinations of crosses between the different necrotic and non‐necrotically reacting genotypes, segregation for the necrotic response in F₂ progeny plants always fitted a 3:1 ratio (necrotic: non‐necrotic). All F₂ progeny plants from the cross between the two non‐cultivar genotypes became infected without necrosis while 99% of the F₂ from the cross between the two cultivars developed necrosis. These results indicate that the systemic necrotic response to infection with BYMV‐N is probably controlled by a single dominant hypersensitivity gene for which we propose the name Nbm‐1. However, its expression seemed influenced by independently segregating modifier genes in the genetic background since necrosis developed at widely different rates within affected F₂ progeny plants resulting in staggered killing.