基于DNA条形码的向日葵田弯管列当种群遗传多样性研究

弯管列当是危害我国向日葵最严重的寄生杂草。为了明确我国向日葵田弯管列当种群的遗传多样性,本试验利用rbcL、matK、ITS2条形码序列对采自我国向日葵主产区的58份弯管列当样品进行PCR扩增及测序,采用Vector NTI软件对测序结果进行剪切比对,利用MEGA 6.0软件计算种内遗传距离并构建系统发育树。利用扫描电镜观察其种子的显微形态特征。结果表明,3个DNA条形码序列中仅ITS2片段扩增测序结果理想并表现出较好的聚类结果。各样品ITS2序列剪切比对后长度为453 bp,种内遗传距离为0.002～0.007,通过比较各样品ITS2序列的碱基组成和差异位点,能将不同弯管列当种群区分开。ITS2聚类结果表明58份弯管列当样品聚为3类,分别为Ⅰ型、Ⅱ型和Ⅲ型。形态分类结果表明,不同类型弯管列当在植株形态、种子形状及微观形态结构等方面存在差别。由于不同弯管列当种群的生境、寄主(向日葵栽培品种)不同,其种群遗传进化差异显著。基于ITS2条形码和扫描电镜形态学观察相结合的方法可用于弯管列当种群遗传多样性研究。     

Proteomics: a promising approach to study biotic interaction in legumes. A review

Summary During the 1990s and early 2000s, the genomes of different organisms have been completely sequenced. Nowadays, biological research is directed to understand gene expression and function. Proteomics, understood as protein biochemistry on an unprecedented and high-throughput scale, is becoming a promising and active approach in this post-genomic period. However, its application to plants is still rather limited as compared to other biological systems. After having referred to the most recent plant proteomic reviews, we focused on legume proteomics including studies with the model species Medicago truncatula. This review is aimed at providing to non-proteomic specialists a global overview of what might be expected in entering this field.     

Inheritance of resistance to race F of broomrape in sunflower lines of different origins

A new race F of broomrape overcomes all known resistance genes in cultivated sunflower, but recently, sources of resistance against race F have been developed. The objective of the present research was to study the inheritance of resistance to race F in crosses between 12 resistant sunflower breeding lines, derived from three different sources of resistance, and the susceptible male‐sterile line P‐21. Parental lines and F₁, F₂, F₃ and BC₁ generations were evaluated for broomrape resistance. Segregations in the F₂ and BC₁ to resistant parent approached resistant to susceptible ratios of 1: 15 and 1: 3, respectively, in most of the crosses, suggesting a double dominant epistasis. However, segregations of 3: 13 and 1: 1 for F₂ and BC₁, respectively, indicating a dominant‐recessive epistasis, were also found. The F₃ data confirmed these results. Owing to the recessive nature of this resistance, it must be incorporated into both parental lines for developing resistant hybrid cultivars.     

Evaluation of Differential Lines and a Collection of Sunflower Parental Lines for Resistance to Broomrape (Orobanche cernua) in Spain

Broomrape (Orobanche cernua Loelf.) is a sunflower parasite which has become a potential threat to this crop in Spam with an increasing incidence in recent last years. In order to investigate the racial situation of the parasite in Spam and to evaluate the resistance of differentials and inbred lines currently used in breeding programmes, two experiments were carried out. In the first, two sets of European sunflower differentials were inoculated with three broomrape populations from different geographical origin. The second experiment studied the reaction to the parasite of 38 inbred lines in comparison with resistant and susceptible checks. Reactions of differential lines resistant to known European broomrape races ranged from complete immunity of lines P-1380-2A and ‘Jdanovski 8281’ to complete susceptibility of ‘Record’ and the universal suscepts. These results confirmed the existence of several physiological races of broomrape in Spain and a different racial composition from that of Eastern Europe. Results from the second experiment showed good resistance of half the restorer lines tested and 36 % of the maintainers and cms lines. This material together with the resistant differential lines constitutes good starting material to be used in developing cultivars resistant to broomrape populations in Spain.     

ISSR标记揭示我国向日葵列当群体的遗传多样性

向日葵列当是目前严重危害我国向日葵生产的寄生性种子植物。为了明确我国不同地区向日葵列当的亲缘关系,本试验利用ISSR标记对采自我国不同省份的96份向日葵列当样本进行了群体遗传多样性的研究。结果表明：从100条ISSR引物中筛选出多态性高、重复性好的12条引物用于群体遗传分析;以供试的列当DNA为模板,利用上述筛选到的引物进行PCR 扩增,共扩增出147 条带,其中90 条具有多态性,多态性条带的百分率为61.2%。不同地区采集的列当群体的Shannon信息指数和Nei′s多样性指数均随着群体样本数量的增加而增大,其中内蒙古地区和新疆地区向日葵列当的多态性相对较高,Shannon信息指数分别为0.5560和0.5067。遗传聚类结果表明6个不同省区的向日葵列当可被聚成两个亚组,其中山西、河北和陕西聚成一个亚组,吉林、新疆和内蒙古聚成另外一个亚组;河北和陕西来源的列当样本亲缘关系最近,而新疆与河北两地的列当样本亲缘关系最远。     

Identification,characterisation and discriminatory power of microsatellite markers in the parasitic weed Orobanche cumana

Orobanche cumana is an obligate root parasite of sunflower. It represents a major agricultural problem in many countries of southern and eastern Europe. Information on O. cumana population genetics, structure and dynamics is scarce, particularly due to the lack of suitable molecular markers for such studies. The objective of this study was to identify and characterise simple sequence repeat (SSR) markers for O. cumana. Four thousand two hundred SSR‐containing candidate sequences were obtained from O. cumana using next‐generation sequencing, from which 298 SSR primer pairs were designed and 217 of them used for validation. Seventy nine SSR primers produced reproducible, high quality amplicons of the expected size that were polymorphic among 18 O. cumana populations from different geographical locations and hosts (sunflower, wild hosts from the Compositae family). The number of alleles per locus ranged from 2 to 10, with an average polymorphism information content value of 0.37. The O. cumana SSR markers were highly transferable to the closely related species Orobanche cernua. SSR markers showed high resolving power; UPGMA cluster analysis allowed proper classification of Orobanche spp. samples into species (O. cumana and O. cernua), geographical origin and host. The functional SSR markers reported in this study constitute a valuable tool for genetic analyses in O. cumana and related species and will contribute insights into the biology and genetics of this parasitic weed.     

Interactions of sunflower (Helianthus annuus) and sunflower broomrape (Orobanche cumana) as affected by sowing date,resource supply and infestation level

The holoparasitic weed Orobanche cumana (sunflower broomrape) constrains sunflower (Helianthus annuus) production in many countries. The development of efficient control strategies requires an understanding of the processes underlying the complex environment–host–parasite interrelations. Growth and development of O. cumana and sunflower were quantified under field conditions in southeastern Romania. Sunflower hybrid Florom 350 was sown at two dates, in plots infested with 0, 50, 200 and 1600 viable O. cumana seeds kg⁻¹ dry soil, under low-input (rainfed, low nitrogen supply) and high-input (irrigated, high nitrogen supply) conditions. Sunflower shoot biomass reached peak values of 760–1287 g m⁻² between the end of anthesis and physiological maturity. Seed yield varied from 221 to 446 g m⁻². Sunflower biomass and yield were affected by all experimental factors. Seed yield responded positively to delaying sowing from early April to late May as well as to irrigation and fertilisation, and negatively to O. cumana infestation. Yield reductions, which were a product of reduced seed number and size, amounted to 13%, 25% and 37% at parasite seed densities of 50, 200 and 1600 viable seeds kg⁻¹ soil, respectively. Maximum O. cumana attachment numbers, recorded in late-sown high-input crops in 2004, ranged from 11 m⁻² in plots with 50 parasite seeds kg⁻¹ soil to 188 m⁻² with 1600 seeds kg⁻¹ soil. Parasite attachment number was a function of crop sowing date, water and nutrient supply, seedbank density, and sunflower biomass and root length density, via mechanisms of parasite seed stimulation, host carrying capacity and intraspecific competition. Delayed sowing and improved water and nitrogen supply were associated with increases in parasite number that neutralised yield-boosting effects of irrigation and fertilisation at the highest infestation level. Sunflower shoot biomass was significantly reduced by O. cumana infection, with reductions affecting organs in the order head > stem > leaves. Most of the discrepancy between infected and non-infected plants was accounted for by O. cumana biomass. Parasites mainly acted as an extra sink for assimilates during sunflower generative growth and impaired host photosynthesis to a much lesser degree. Results suggest that similar mechanisms govern infection level and host–parasite biomass partitioning across different Orobanche–host systems.     

列当(Orobanche spp. and Phelipanche spp.)种子的采集与预处理方法

列当(Orobanche spp.and Phelipanche spp.)是一种在世界范围内危害严重的根寄生杂草,防除列当的研究也越来越多。正确地采集和预培养列当种子是获得正确防除列当研究结果的前提。列出了大田正确采集列当种子的方法,以及对采集到的种子进行过筛提纯、加洗洁精和吐温20清洗后,再表面消毒和预培养,并通过发芽试验检验此方法的可行性。结果表明,经过处理的向日葵列当、瓜列当种子的发芽率最高分别为54.4%、59.1%。此方法可以应用于列当的采集和预处理过程中,可为列当研究和防除提供参考。     

Simple sequence repeats (SSRs) in faba bean: new loci from Orobanche-resistant cultivar 'Giza 402'

Genomic DNA from an inbred line from the Orobanche crenata resistant cultivar 'Giza 402' was restricted and enriched for AG-containing simple sequence repeats (SSRs). The overall SSR enrichment rate was 54.6%. Of 73 selected SSR markers that successfully amplified in 'Giza 402' 54 were polymorphic among a group of 10 Egyptian and Spanish faba bean genotypes extensively used in Orobanche crenata resistance/susceptibility studies and linkage map development. The polymorphism information content ranged from 0.16 to 0.72 and Unweighted Pair-Group Method using Arithmetic averages cluster analysis based on these loci placed all resistant genotypes descended from the parental line F 402 in the same group. Successful amplification from 28 primer pairs was also observed in five European faba bean genotypes. The SSR loci described here will be extremely useful in studies focusing on Orobanche crenata resistance, genetic diversity studies and will provide additional co-dominant loci to complement existing faba bean linkage maps based primarily on dominant markers.     

不同年代冬小麦品种根际土浸提液诱导小列当种子发芽的化感作用研究

为筛选在根寄生植物小列当多发地进行诱捕发芽的小麦品种,研究了不同品种冬小麦根分泌的化感物质对小列当(Orobanche minor)种子的诱导发芽率。以冬小麦根际土蒸馏水和甲醇浸提液刺激小列当种子发芽,结果表明,冬小麦品种间化感作用差异显著。20世纪70年代宁冬1号和90年代小偃22号在4个生育期都有较强的发芽刺激作用,而其余3个品种则在个别生育期有非常微弱化感作用。宁冬1号品种刺激小列当发芽率达到最高为42.8%,化感作用最强,可作为抑除小列当的首选品种。