QTL mapping and associated marker selection for the efficacy of green plant regeneration in anther culture of rice

Anther culturability of rice is a quantitative trait controlled by nuclear‐encoded genes. The identification of quantitative trait loci (QTL) and associated marker selection for anther culturability is important for increasing the efficiency of green plant regeneration from microspores. QTL associated with the capacity for green plant regeneration in anther culture of rice were mapped on chromosomes 3 and 10 using 164 recombinant inbred (RI) lines from a cross between ‘Milyang 23’ and ‘Gihobyeo’. The quantitative trait locus located on chromosome 10 was detected repeatedly when three anther culture methods were applied and was tightly linked to the markers, RG323, RG241 and RZ400. Associations between these markers and the efficacy of green plant regeneration in 43 rice cultivars and two F₂ populations, ‘MG RI036’/‘Milyang 23’, and ‘MG RI036’;/‘IR 36’ were analysed. One of these markers, RZ400, was able to identify effectively genotypes with good (> 10.0%) and poor (< 3.0%) regenerability, based on the marker genotypes in the cultivars and two F₂ populations. This marker enables the screening of rice germplasm for anther culturability and introgression into elite lines in breeding programmes.     

Homozygous transgenic rice lines expressing GNA with enhanced resistance to the rice sap-sucking pest Laodelphax striatellus

Homozygous transgenic rice lines expressing snowdrop lectin, Galanthus nivalis agglutinin (GNA), were investigated for resistance to the rice small brown planthopper (SBPH), Laodelphax striatellus, an important rice sap‐sucking pest causing yield losses and serving as vector for some important rice viruses. Insect bioassay results revealed that both the GNA‐expressing homozygous lines tested significantly inhibited SBPH by decreasing its survival, its overall fecundity and retarding development. This is the first report on homozygous transgenic rice lines expressing GNA conferring enhanced resistance to SBPH. This result suggests that transgenic rice expressing GNA will be useful in rice pest resistance breeding, as an alternative to conventional breeding for the control of SBPH, and potentially alleviating damage caused by the viruses it transmits.     

Homoeological relationships between the f chromosome of Brassica rapa and the e chromosome of Brassica oleracea

Eight plants of the putative double monosomic addition line (DMAL, 2n= 20) were developed by crossing a monosomic chromosome addition line of radish [f(A)‐type monosomic addition line (MAL) (2n= 19)] carrying the f chromosome of Brassica rapa (2n= 20, AA) with another [e(C)‐type MAL (2n= 19)] having the echromosome of Brassica oleracea (2n= 18, CC). The homoeological relationships between the two alien chromosomes were investigated by morphological, cytogenetic and random amplified polymorphic DNA (RAPD) analysis. Seventeen morphological traits that were not present in the radish cv. ‘Shogoin’ were observed in both MALs and these traits were substantially exhibited in DMAL plants. At the first metaphase of pollen mother cells (PMCs), the two parental MALs showed a chromosome configuration of 9II +1I, demonstrating impossibility of recombination between the R and the added chromosomes. The DMALs formed 10II in approximately 73% of PMCs, with one bivalent showing loose pairing between two chromosomes differing in size. In an attempt to identify the two MALs by RAPD‐specific markers using 26 selected random primers, 13 and 20 bands were specific for the f(A)‐type and the e(C)‐type MALs, respectively; 12 bands were common to both MALs (26.7%). In conclusion, the f chromosome of B. rapa is homoeologous to the e chromosome of B. oleracea. The genetic domain (genes) for 17 morphological traits are linked to each homoeologous chromosome bearing 27% of the corresponding RAPD markers.     

Breeding maize lines for complete and partial resistance to maize streak virus (MSV)

Summary S₁ to S₅ inbred lines, derived from a maize population bred for its overall resistance to three tropical viruses, were screened for resistance to maize streak virus (MSV) by artificial plant infection using viruliferous leafhoppers. Symptoms were rated and intra-line frequency distributions studied for all pedigree inbred lines. Mortality due to MSV was very low among these inbreds. Symptoms appeared later, developed slower and were less severe than in the susceptible control hybrid. Results of a study of 500 S₁ and 93 S₂ lines suggested that resistance is under genetic control via a system involving loci with major genes (with dominance for resistance) controlling high to complete resistance, associated with a genetic system involving loci with minor genes controlling partial resistance. Lines expressing complete resistance to MSV were developed from 5 cycles of inbreeding and selection. The relevance of such complete and partial resistance is discussed.Abbreviations MRPS Mean Rating for Plants exhibiting Symptoms     

Identification and Inheritance of Fertility Restorer Genes for 'tour' CMS in Rapeseed (Brassica napus L.)

Eighteen genotypes of Brassica napus were crossed to a cytoplasmic male sterile (CMS) line of B. napus BO 15 carrying B. tournefortii cytoplasm (‘tour’ cytoplasm). Fourteen genotypes were found to be stable maintainers of the ‘tour’ CMS. Of the remaining four genotypes, GSL-1 and ‘Asahi-natane’ were found to be heterozygous and ‘Mangun’ and ‘Yudal’ were homozygous for the restorer gene. Analysis of the F₁ and F₂ progenies of (CMS) BO 15 בMangun’ and (CMS) BO 15 בYudal’ showed that fertility restoration is controlled by a single dominant gene. The availability of a number of stable maintainer lines and the simple inheritance pattern of fertility restorer gene makes ‘tour’ CMS a useful system for hybrid seed production in rapeseed.     

Limited but specific variations of seed storage proteins in Japanese common wheat (Triticum aestivum L.)

The electrophoretic banding patterns ofgliadin in common wheat lines derived fromJapan were determined byacid-polyacrylamide gel electrophoresis. For the 107 wheat lines used in our study,27 different patterns were identified, 13corresponding to the -gliadin, 8 tothe , -gliadin and 6 to the-gliadin. The gliadin patterns ofJapanese wheat cultivars and landracesgreatly differed from the patterns of wheatlines from other countries, and thevariation seen in wheat lines from Japanwas limited to 46 patterns. Sevencollection or breeding areas in Japanshowed different frequencies in theirgliadin patterns. Combining the gliadinpatterns with high molecular weightglutenin subunit compositions, 67combinations were observed. One gliadinpattern consisting of -gliadinpattern F, , -gliadinpattern H and -gliadin pattern Dwas frequently found in many Japanese wheatlines, though the other patterns werelimited to only one or two wheat lines.     

Field evaluation of insect-resistant transgenic Populus nigra trees

The performance of insect-resistant transgenic poplar trees (Populusnigra) expressing a Cry1Ac gene from Bacillus thuringiensis subsp. Kurstaki HD-1 against poplar defoliators was evaluated in the field at the Manas Forest Station in Xinjiang Uygur Autonomous Region during1994–1997. The results showed that the average percentage of highly damaged leaves on the transgenic trees was 10% while that on the control trees in nearby plantations reached 80–90%. The average number of pupae per m² of soil at 20cm depth in transgenic poplar plantation was 18 which was only 20% of that found in the non-transgenic control field. The number of pupae and the leaf-damage on transgenic trees described above are all far below the threshold set for chemical protection measures. The non-transformed poplar trees grown in the same plantation with the transgenic trees were also protected indicating that cross protection occurred between these two kinds of trees. Insect-resistant transgenic poplar trees have a potential application value in afforestation. This revised version was published online in August 2006 with corrections to the Cover Date.     

Factors influencing tissue culture responses of mature seeds and immature embryos in turf-type tall fescue

In an effort to optimize tissue culture responses of turf‐type tall fescue for genetic transformation, the effects of five culture medium supplements on tissue culture responses were investigated with immature embryos and mature seeds of an elite cultivar, ‘Coronado’, as explant tissues. For both explants, calli induced on 6‐benzylaminopurine (BAP)‐containing medium had significantly improved regeneration ability. The optimal concentration of BAP for the induction of regenerable callus from mature seeds was 0.1 mg/l. Thidiazuron also improved callus regeneration frequency of mature seeds. Casein hydrolysate, L‐proline and myo‐inositol improved callus induction of immature embryos but not mature seeds. They did not improve callus regeneration frequencies with either explant tissue. By simply slicing the mature seeds into two halves longitudinally, the callus induction frequencies, as well as the corresponding overall plant regeneration frequencies, were increased approximately three‐ to six‐fold in all three combinations of 2,4‐dichlorophenoxyacetic acid and BAP in callus induction media.     

Genetic analysis of presence and absence of lint and fuzz in cotton

Cotton fibre mutants that were fuzzless and/or lintless were crossed with each other and a normal genotype (fuzzy, linted) to produce F₂ and BC₁ generations. F₂ segregation ratios from the cross of fuzzless‐lintless × fuzzy‐linted, for fuzzy‐linted, fuzzless‐linted and fuzzless‐lintless were 45 : 15 : 4. From the cross of fuzzless‐lintless × fuzzy‐linted, the F₂ segregation ratios were 9 : 39 : 16 whereas the BC₁F₁ segregation ratios from the F₁ backcrossed to fuzzless‐lintless were 1 : 3 : 4. These data suggest that the presence or absence of lint and fuzz are controlled by the interaction of four gene loci on non‐homologous chromosomes. We designate these loci as N₁, N₂, Li₃ and Li₄, where N₁ N₁ confers the presence of fuzzy, N₂N₂ confers inhibition of fuzzy initiation and development, and duplicate gene pairs, Li₃Li₃ and Li₄Li₄, determine the presence of lint. Homozygosity for li₃li₃ and li₄li₄ might also inhibit fuzz from development. In other words, they were recessive epistatic to fuzz genes.     

A high-resolution map of the rice blast resistance gene Pi15 constructed by sequence-ready markers

The gene Pi15 for resistance of rice to Magnaporthe grisea was previously mapped to a ≈0.7-cM region on chromosome 9. To further define the chromosomal region of the Pi15 locus, a contig spanning the locus was constructed, in silico , through bioinformatics analysis using a reference sequence of the cultivar 'Nipponbare'. One simple sequence repeat marker adopted from the International Rice Microsatellite Initiative and six candidate resistance gene (CRG) markers, developed from gene annotation of the reference sequence of the contig, were used for linkage analysis in a mapping population consisting of 504 extremely susceptible F₂ plants. The Pi15 locus was delimited to a ≈0.5-cM region flanked by the markers CRG5 and CRG2 and co-segregated with the markers BAPi15₇₈₂, CRG3 and CRG4, which was physically converted to a 44-kb interval.