Congruence of conventional and molecular studies to locate genes that control flavone synthesis in maize silks

The presence of maysin, a flavone glycoside, and its analogues in the silks of corn is an important defence against invasion of the ear by corn earworm in the southeastern USA. Three dent maize inbreds with high silk‐maysin concentrations were evaluated for chromosomal location of major genes controlling synthesis of three antibiotic chemicals by crossing them to a series of waxy‐marked chromosome 9 reciprocal translocations. The data indicated that genes for maysin and its analogues are probably present on the short arms of chromosomes 1 and 10, and long arms of chromosomes 5 and 8 for inbred line GT114, the short arms of chromosome 1 and 6, and the long arms of chromosome 5 and 8 for inbred line GE37, and the short arms of chromosomes 1 and 10, and the long arm of chromosome 4 for inbred line SC102. These results are in general agreement with other translocation studies on corn earworm injury to sweet corn inbreds and gene and restriction fragment length polymorphism marker studies to locate quantitative trait loci (QTL) for maysin synthesis, with the exception that QTL on chromosome 9 have been found to be strongly associated with flavone synthesis. The most significant concordance between conventional and molecular techniques for locating chromosome regions influencing synthesis of antibiotic chemicals from silks of high silk antibiosis sources is found for the short arm of chromosome 1. This region is the most likely one on which to place emphasis during the initial stages of transferring high‐maysin silks to elite inbred lines. A chromosome 1 marker plus at least one more from any chosen high‐maysin inbred source should be sufficient to transfer high maysin silks to other lines. Other markers may be needed for transfer of specific traits when recovering recurrent parent genome types in a backcrossing procedure.     

基于寡核苷酸探针套painting的中国春非整倍体高清核型及应用

基于寡核苷酸探针套painting的染色体鉴定技术简单、经济和高效, 可以促进小麦品种及亲缘物种染色体识别和变异体鉴定, 提高染色体工程效率。我们前期开发了寡核苷酸探针套, 包含pAs1-1、pAs1-3、AFA-4、(GAA)₁₀和pSc119.2-1共5个探针。本研究通过一次荧光原位杂交(FISH), 对源于17个非整倍体的18份材料分析发现, 其中14个染色体组成正确, 可以清晰识别相应的缺体、四体和端体。还构建了基于寡核苷酸探针套涂染的、能准确识别3个基因组和7个部分同源群染色体的高清核型, 发现4个非整倍体发生变异, 其中从N5BT5D中鉴定出一个可能的小片段相互易位系T6AS·6AL-6DL和T6DS·6DL-6AL。进一步对7个地方品种、10个栽培品种(系)和1个人工合成小麦分析, 发现15条染色体存在多态性, 涉及6条B组(除4B)、5条A组(除1A和3A)和4条D组(1D、2D、4D和7D)染色体, 可以清晰识别我国小麦生产上广泛应用的3种易位类型(T1RS·1BL、T6VS·6AL及相互易位T1RS·7DL和T7DS·1BL), 省去了基因组原位杂交(GISH)程序。另外, 对5个亲缘物种分析发现, 该探针套可以识别栽培一粒小麦、硬粒小麦Langdon、荆州黑麦、长穗偃麦草(2n=2x=14)全部和中间偃麦草30条染色体, 并构建了这5个物种的核型。本研究结果证实该寡核苷酸探针套可以有效用于小麦及亲缘物种染色体鉴定, 高清晰的中国春非整倍体核型为小麦染色体工程提供了参考标准。     

Wildlife diseases in New Zealand: recent findings and future challenges

Our knowledge of diseases in New Zealand wildlife has expanded rapidly in the last two decades. Much of this is due to a greater awareness of disease as a cause of mortality in some of our highly threatened species or as a limiting factor to the successful captive rearing of intensely managed species such as hihi (Notiomystis cincta), kiwi (Apteryx spp.) and kakapo (Strigops habroptilus). An important factor contributing to the increase of our knowledge has been the development of new diagnostic techniques in the fields of molecular biology and immunohistochemistry, particularly for the diagnosis and epidemiology of viral and protozoan diseases. Although New Zealand remains free of serious exotic viruses there has been much work on understanding the taxonomy and epidemiology of local strains of avipox virus and circoviruses. Bacterial diseases such as salmonellosis, erysipelas and tuberculosis have also been closely investigated in wildlife and opportunist mycotic infections such as aspergillosis remain a major problem in many species. Nutritional diseases such as hyperplastic goitre due to iodine deficiency and metabolic bone disease due to Ca:P imbalance have made significant impacts on some captive reared birds, while lead poisoning is a problem in some localities. The increasing use of wildlife translocations to avoid the extinction of threatened species has highlighted the need for improved methods to assess the disease risks inherent in these operations and other intensive conservation management strategies such as creching young animals. We have also become more aware of the likelihood of inbreeding suppression as populations of many species decrease or pass through a genetic bottleneck. Climate change and habitat loss, however, remain the greatest threats to biodiversity and wildlife health worldwide. Temperature changes will affect our wildlife habitats, alter the distribution of disease vectors and wildlife predators, or directly harm threatened species in vulnerable localities.     

寡核苷酸探针套涂染结合基因组原位杂交和分子标记分析准确鉴定小麦-百萨偃麦草异源易位系的研究

培育小麦异源易位系是转移和利用外源基因的有效途径,快速准确的染色体鉴定方法有利于提高染色体工程效率。本研究以电离辐射诱致的2个普通小麦-百萨偃麦草染色体5J易位系（NAU16YJ127和NAU16YJ124）为例,分析了综合利用本课题组前期开发的寡核苷酸探针套［包括(GAA)₁₀、pSc119.2-1、pAs1-1、pAs1-3、AfA-3和AfA-4六个寡核苷酸探针］涂染,结合基因组原位杂交和分子标记等方法鉴定小麦异源易位系的效果。结果表明,NAU16YJ127（2n=6x=42）包含一对小麦-百萨偃麦草易位染色体T5JS·5JL-5AL#1,同时在染色体6A和2D长臂顶端也发生了明显的相互易位;NAU16YJ124（2n=6x=44）附加了一对小麦-百萨偃麦草易位染色体,同时包含一对6A长臂缺失染色体del6AL,其缺失区段易位到5JL近端部,形成T5JS·5JL-6AL,其他染色体未见明显变化。利用21个5J特异分子标记（19个位于长臂和2个位于短臂）分析发现,两个易位系均含有全部5J短臂标记,同时T5JS·5JL-5AL#1含有12个长臂标记,而T5JS·5JL-6AL仅含有2个长臂标记,确证两个易位系断点不同,并将21个标记定位于4个不同区段,其中位于5J短臂的2个标记位于1个区段,位于5J长臂的19个标记位于3个区段（5JL-1~3）,分别包含2、10和7个标记;发现3个为可以同时识别5J和5A的共显性标记,其中NAU16YJ124含有全部5A标记,而NAU16YJ127仅包含其中2个标记,进一步验证了细胞学鉴定结果。本研究结果表明,寡核苷酸探针套涂染、基因组原位杂交和分子标记分析相结合为准确鉴定小麦异源易位系提供了新方法。     

Network analysis of wildlife translocations in New Zealand

AIMS: To identify network measures with relevance to disease spread in a network of movements derived from the Department of Conservation (DOC) translocation records from 1970 to mid-2014, and to identify conservation sites that should be prioritised for surveillance activities and improvements to data collection to make the best use of network analysis techniques in the future.

METHODS: Data included the source and destination of translocated specimens, the species and the dates the translocations were expected to occur. The data were used to construct a directed, non-weighted network in which a translocation event represented a tie in the network. Network density, in-degree (movements entering a node of interest) and out-degree (movements leaving a node of interest) and reciprocity were calculated.

RESULTS: The data analysed consisted of 692 unique translocations between 307 sites, with the majority (518; 73%) being for birds. The constructed network for bird, reptile and frog translocations comprised 260 nodes, with 34/260 (13%) having two-way movements and 47/260 (18%) non-reciprocal movements. The median degree score (sum of in- and out-degree) was two (min 0, max 36) with a mean of 3.5 in a right skewed distribution. Most sites acted as receivers or senders of consignments with only a few having both high in- and high out-degree, and thus had characteristics that made them sites of interest for surveillance activities. These included the National Wildlife Centre at Mount Bruce, Tiritiri Matangi Island and Te Kakahu (Chalky Island).

CONCLUSIONS: The presence of linking sites that join larger clusters within the network creates the potential for rapid disease spread if a pathogen were to be introduced. The important sites that supply or receive specimens for translocations are already well recognised by those performing translocations in New Zealand, and this paper provides further information by quantifying their role within the network.     

An STS marker distinguishing the rye-derived powdery mildew resistance alleles at the Pm8/Pm17 locus of common wheat

A sequence‐tagged site marker has been developed from restriction fragment length polymorphism marker probe IAG95 for the rye‐derived powdery mildew resistance Pm8/Pm17 locus of common wheat. This polymerase chain reaction marker enables the amplification of DNA fragments with different sizes from T1AL.1RS and T1BL.1RS wheat‐rye translocation cultivars with chromatin from ‘Insave’ and ‘Petkus’ rye, respectively, and therefore will be very useful in distinguishing Pm8‐carrying cultivars from Pm17‐carrying cultivars. Results obtained with that marker were compared with resistance tests performed on detached primary leaves of 29 wheat lines from two populations derived from doubled haploid production. The molecular assay corresponded well with the resistance tests in all the lines, and therefore will be helpful for the identification of Pm17 in lines in which other Pm genes or quantitative trait loci are present.     

Genetic variation in germplasm accessions of Secale vavilovii Grossh

Summary The accurate identification and classification of wild species enhance the utilization of potential plant genetic resources. The wild rye species Secale vavilovii Grossh. may serve to broaden the genetic variation in cultivated rye, S. cereale. The combined analyses of cytological, isozymic and other genetic characteristics in the present study revealed that of the fourteen rye accessions designated as S. vavilovii, only three accessions were correctly classified. Thus it is essential to increase multidisciplinary, international research and collaboration to aid in the classification and utilization of the yet untapped plant germplasm.     

Genomic constitution of Festulolium cultivars released in the Czech Republic

Genomic in situ hybridization (GISH) was used to characterize the chromosome constitutions of individual plants from a set of tetraploid and hexaploid cultivars of Festulolium developed and released in the Czech Republic from hybrids of Lolium multiflorum with Festuca pratensis and F. arundinacea. A simplified GISH protocol readily discriminated parental genomes in the hybrids and facilitated the screening of large numbers of plants per accession. The contribution of parental genomes in the cultivars tested ranged from predominance of chromatin from one of the parents to a more balanced contribution from both parents. However, in none of the cultivars were equal proportions of chromatin from both parents present. The parental contribution to the hybrids was both in the form of complete chromosomes or as chromosome translocations. In hexaploid cultivars from (L. multiflorum × F. arundinacea) × F. arundinacea hybrids the average numbers of complete L. multiflorum chromosomes ranged from 4.95 to 7.5 and the numbers of translocations from 6.33 to 10.21. Two tetraploid cultivars from (L. multiflorum × F. arundinacea) × L. multiflorum hybrids showed a strong prevalence of L. multiflorum chromatin and intergeneric translocations were rare. In the tetraploid cultivar ‘Perun’ of the L. multiflorum × F. pratensis hybrid there were 11.7 chromosomes of L. multiflorum and 14.7 recombined chromosomes on average. Reasons for the domination of one of the parental genomes in hybrid cultivars are not clear and are only partially explained by breeding history. Recombination rates of individual genomes in hybrids involving F. arundinacea were evaluated in double hybridization experiments. The results indicated a strong affinity of the L. multiflorum genome for the F. pratensis genome present in F. arundinacea and little affinity for the F. glaucescens genome. This suggests that introgressions from F. arundinacea into L. multiflorum are primarily limited to the F. pratensis genome which can be more readily accessed in L. multiflorum × F. pratensis hybrids.     

Identification of Rye Chromosome 1R Translocations and Substitutions in Hexaploid Wheats Using Storage Proteins as Genetic Markers

Grain protein compositions of 106 advanced generation backcross lines from crosses involving ‘Amigo’ (1AL.1RS), ‘Aurora’, ‘Kavkaz’, ‘Skorospelka-35’ and ‘Sunbird’ (all 1BL.1RS) and ‘Gabo’ 1DL.1RS parents and 152 cultivars with unknown pedigree were analysed by one-dimensional SDS-PAGE. Eighty seven backcross lines and 16 cultivars carried one or other of these translocations, 2 cultivars had a 1R (1B) substitution, whereas 5 backcross lines were found to be heterogeneous for the 1BL.1RS translocation. The translocation lines were easily identified by the presence of secalins (Sec-1) controlled by rye chromosome arm IRS and a simultaneous loss of the gliadin (Gli-1) and/or triticin (Tri-1) protein bands controlled by the replaced wheat chromosome arm (1AS, 1BS or 1DS). Certain gliadins, showing no allelic variation among the genotypes analysed, were identified as markers for chromosome arms 1AS (M_r= 34 kd) and IBS (M_r= 42,33 kd). The whole chromosome substitutions 1R (1B) were recognized by scoring for the presence of Sec-1 and HMW secalin bands, Sec-3 (controlled by rye chromosome arm 1RL) and the absence of Gli-B1 and HMW glutenin subunits, Glu-B1 (controlled by wheat chromosome arm 1BL). The results have shown that protein electrophoresis provides a rapid and reliable technique for screening genotypes for these translocations and substitutions in a breeding programme.