Two major genes for kernel weight in rye

Previously, kernel weight in rye was considered mostly as a quantitatively inherited trait. F₁ plants from parents showing a great difference in kernel weight were selfed and the segregating F₂ population was analysed with molecular markers. Two simple sequence repeats markers, one on chromosome 5 and the other on chromosome 7, were found which allowed the genetic analysis of two major genes acting in a complementary way. The analysis of major genes is an advantage in breeding for this important economic trait. In addition it can provide more insight into the genetic structure of this character. This is, in turn, also a prerequisite for future investigations of this trait at the molecular level.     

Isolation of a new repetitive DNA sequence from Secale africanum enables targeting of Secale chromatin in wheat background

A genome specific DNA sequence that detects Secale africanum chromatin incorporated into wheat was developed in this study. Random amplified polymorphic DNA (RAPD) analysis was used to search for genome specific DNA sequences of S. africanum in lines, R111, “mianyang11” (MY11) and wheat-rye 1RS/1BL translocations R25 and R57. A high copy rye-specific DNA segment pSaD15₉₄₀ of the S. africanum genome was obtained. The sequence of pSaD15 did not show any significant homology to other reported sequences in databases and it is therefore a new repetitive sequence of Secale. PCR primers were designed for pSaD15₉₄₀, which amplify a clear 887 bp fragment in S. africanum but not in any wheat. The primers also amplified an 887 bp fragment in other accessions of rye, Chinese Spring-Imperial rye chromosome additions and a diverse range of material carrying different rye chromosomes or chromosomal segments. In situ hybridization showed that probe pSaD15₉₄₀ was specifically hybridized throughout all rye chromosomes arms except for the terminal regions. The advantage of the rye-specific probe developed herein compared to those of previous reports is that it has been shown to be widely applicable to other Secale species. The probe will be useful as a molecular marker for the introgression of S. africanum and other rye chromosome segments into the wheat genome.     

Expression of nucleolus, endosperm storage proteins and disease resistance in an amphiploid between Aegilops tauschii and Secale silvestre

Cytology and gene expression of an amphiploid between Aegilops tauschiiL., native to China, and Secale silvestre L. were studied to reveal the genomic interaction between the donor species. High frequencies of aneuploids were observed in the progenies of the amphiploid, indicating its cytological instability. Feulgen staining and Giemsa-C banding showed that only the nucleolar organizing region from chromosome 5D of Ae. tauschii existed in the amphiploid (2n = 28). The nucleolus of S. silvestre was not observed. Endosperm storage protein electrophoresis indicated most gliadin and glutenin genes from both parents were expressed in the endosperm of the amphiploid. When inoculated by wheat stripe rust and powdery mildew isolates,the amphiploid did not express the resistance from its Secale parent,suggesting the presence of disease resistance suppressor(s) in the D genome of Ae. tauschii as well as nucleolar organizer suppressors. This revised version was published online in July 2006 with corrections to the Cover Date.     

Rye introgression lines as source of alleles for pollen-fertility restoration in Pampa CMS

Restoration of pollen-fertility is required to guarantee successful seed production of hybrid varieties based on cytoplasmatic male-sterility (CMS) induced by Pampa cytoplasm. As most European restorer lines have still numerous disadvantages, using genetic resources via introgression libraries might be promising to identify and localize new and more effective restorer genes. The objectives of this study were to (1) evaluate in field trials the performance of 39 candidate introgression lines (pre-ILs) for pollen-fertility restoration and (2) to identify donor chromosome segments (DCS) underlying pollen-fertility restoration. Exotic alleles from the Iranian primitive rye accession Altevogt 14160 were introduced into the elite non-restorer inbred line L2053-N by marker-assisted backcrossing. BC₂S₃ pre-ILs were genotyped using amplified fragment length polymorphisms and simple sequence repeats. Pollen-fertility restoration was evaluated in replicated field trials across two testers at two locations in 2006 for analysing the phenotypic effects of the exotic DCS. Out of 39 pre-ILs, we determined three showing full pollen-fertility across both testers and locations carrying a DCS on chromosome 4R. Moreover, one pre-IL with partial pollen-fertility was detected showing a DCS on chromosome 1R. We conclude that our introgression library can serve as a valuable resource for mining favourable genes from exotic primitive rye, which can greatly promote hybrid rye breeding.     

普通小麦辉县红-荆州黑麦异染色体系的选育及其梭条花叶病抗性鉴定

为发掘和利用荆州黑麦所携抗梭条花叶病基因，综合利用分子细胞遗传学与分子标记技术结合多年抗性鉴定，从高感梭条花叶病小麦地方品种辉县红与荆州黑麦杂交后代（F₇~F₉）中选育出二体异附加系5个(分别添加1R、2R、R3、5R和R7)、5RS端二体异附加系1个和多重异附加代换系2个（染色体组成分别为20’’+2R(2D)’’+4R’’和19’’+1R（1B）’’+2R（2B）’’+4R’’）。鉴定表明，双二倍体荆辉1号高抗梭条花叶病，表明黑麦抗性基因可在小麦背景中稳定表达，2R、R7二体异附加系及2个含2R的多重异附加代换系均表现高抗，推测2R和R7上可能携带抗病基因。这些材料是研究荆州黑麦抗性基因遗传及小麦抗病育种的新种质。     

Significance of host complexity and diversity for race-specific leaf-rust resistance in self-fertile synthetic rye populations

Leaf rust (Puccinia recondita Rob. ex. Des.) is the most frequently occurring leaf disease in German winter rye (Secale cereale L.). To test the usefulness of race‐specific resistance genes, the effects of increased host diversity and complexity by producing two‐ and four‐line synthetics from inbred lines carrying different resistance genes were analysed. Thirty‐three synthetics along with two full‐sib families and one hybrid variety were tested in 17 environments in Germany under natural infections. For comparison, the parent lines of the synthetics were evaluated in 11 environments. Only two synthetics and the full‐sib families were resistant across all environments. Observed resistance levels of the synthetics were highly correlated (r = 0.83, P = 0.01) with those predicted from the parental values. Host complexity had a minor effect in two‐line synthetics only. In conclusion, the effectiveness of race‐specific leaf‐rust resistances among environments, and increasing the host complexity and diversity does not lead to a higher resistance level than that expected from the resistances of the parents.     

小麦EST-SSR标记的开发和染色体定位及其在追踪黑麦染色体中的应用

根据小麦盐胁迫诱导和茎秆组织相关EST序列开发了81对EST-SSR引物, 其中67、46、18和61对分别在小麦、黑麦、簇毛麦和大麦基因组中稳定扩增, 在不同小麦和大麦品种间具有多态性的引物分别有22和23对。利用小麦缺体-四体系共定位了43对引物的81个位点, 其中A、B和D染色体组上分别有29、30和22个位点, 涉及除4B、3D和6D外的18条染色体。此外30对引物在黑麦基因组中具有特异扩增, 其中8对分别在黑麦1R、4R、5R和R7染色体上具有特异扩增, 7对在多条黑麦染色体具有相同扩增。这些新标记可有效用于小麦及其近缘物种的遗传作图与比较遗传研究。     

Stability of RAPD markers for determining cultivar specific DNA profiles in rye (Secale cereale L.)

Summary Cultivar specific DNA profiles in rye were revealed by polymerase chain reaction (PCR) using randomly amplified polymorphic DNA (RAPD) sequences. Ten base primers were used for the amplification of genomic DNA of rye cultivars by PCR. RAPD analysis was found to be reproducible among samples between PCR runs. When amplification profiles of different rye cultivars were compared using various primers, the overall profiles were cultivar specific. However, not all primers revealed polymorphisms. These primers appear to amplify conserved sequences in all rye cultivars. Intracultivar studies were conducted on two of the cultivars. In the cultivar Imperial, no polymorphisms were observed among ten plants analyzed with five primers. In the cultivar Balboa, polymorphisms were observed among fifty plants with four of the ten primers analyzed. Despite the small amount of intracultivar variability, RAPD analysis has the potential to be a rapid and reliable method of cultivar identification in this outcrossing species.     

Identification of somatic hybrids of dihaploid Solanum tuberosum lines and S. brevidens by species specific RAPD patterns and assessment of disease resistance of the hybrids

Summary Symmetric somatic hybrids were produced by electrofusion of protoplasts of two dihaploid tuber-bearing potato (Solanum tuberosum L.) lines and Solanum brevidens Phil., a diploid non-tuber-bearing wild potato species. A total of 985 plants was obtained. Verification of nuclear hybridity of putative hybrids was based on additive RAPD patterns, general morphological characteristics and chromosome counts. 53 (90%) calli regenerated into plants which were identified as somatic hybrids. Most of the hybrids were aneuploids at the tetraploid (4×) or hexaploid (6×) level. The 20 hybrids tested expressed a high level of resistance to potato virus Y (PVY ^N ) characteristic of the S. brevidens parent. Resistance to late blight (Phytophthora infestans (Mont.) de Bary) varied between hybrids, but was on average better than that of the fusion parents. Resistance of hybrids to bacterial stem rot (Erwinia carotovora subsp. atroseptica (van Hall) Dye) was not superior to that of commercial potato cultivars.