簇毛麦6VS特异转录序列P21461及P33259的获得及其分子标记在鉴定小麦-簇毛麦抗白粉病育种材料中的应用

簇毛麦6V#2S和6V#4S染色体臂分别携带抗白粉病基因Pm21和Pm V,在与小麦的杂种后代中,抗病基因与外源染色体臂共分离。开发鉴定2条外源染色体臂间多态性的序列,尤其是遗传信息相对缺乏的6V#4S染色体臂的序列,对于其在遗传与育种上的应用具有重要意义。本研究以携带6V#4S·6DL染色体的小麦易位系Pm97033及感病小麦亲本宛7107接种白粉菌的叶片转录组数据为资源,通过差异基因筛选、共线性分析、簇毛麦基因组扩增及测序验证的方法,鉴定出来自6V#4S的表达序列P21461和P33259,其中基于P21461序列设计的引物P461-5在簇毛麦6V#2S和6V#4S染色体臂的扩增产物具有30 bp的In Del和4 nt的多态性。用该引物转化的标记P461-5a可以鉴定抗白粉病小麦品种和高代品系所含的外源染色体,显示其在簇毛麦抗源鉴别和小麦抗病育种辅助选择中潜在的应用价值。根据P33259开发的标记P259-1可以对含有6V#4S染色体臂的材料进行特异扩增,但对6V#2S·6AL易位染色体没有扩增产物,因此P259-1可作为6V#4S·6DL易位染色体的特异分子标记。q RT-PCR分析结果显示,P21461的表达不受白粉菌诱导,而P33259在接菌后12 h和24 h的转录水平比接菌前提高约2倍,推测其可能参与Pm97033与白粉菌的早期互作。     

受辐照窄颖赖草花粉DNA进入小麦胚囊的电镜自显影证据及杂种原位杂交鉴定

采用3H 胸腺嘧啶标记窄颖赖草 (Leymusangustus)花粉结合电镜自显影的方法证明受辐照窄颖赖草花粉DNA进入了小麦胚囊 ,运用基因组原位杂交技术证明所得杂种为真杂种 ,经辐照花粉获得的普通小麦J 1 1与窄颖赖草杂种中窄颖赖草染色体发生了数目和结构变异     

Late nitrogen fertilization affects carbohydrate mobilization in wheat

An experiment was carried out to determine how the late application of nitrogen (N) fertilizer affects the use of pre‐anthesis carbon reserves during the grain‐filling period of pot‐grown wheat with no water shortage. Increasing doses (equivalent to 0, 140, and 180 kg N ha^–1) of N fertilizer were applied, either in two amendments (stages GS20 and GS30) or in three amendments (stages GS20, GS30, and GS37, according to Zadoks scale). The management of fertilizer by combining an increased N rate with late N application was able to stimulate canopy development, to raise photosynthetic capacity and carbohydrate accumulation during the vegetative stages, and to increase grain yield. Based on the dynamics of carbohydrate accumulation in the ear, three phases were differentiated during the grain‐filling period, whose temporary pattern remained stable regardless of the fertilizer management. The net remobilization of carbohydrates started 12 d after anthesis from the leaves and 28 d after anthesis from roots and stems. The increase of the N dose with late N application allowed on one hand a lower use of the pre‐anthesis carbon reserves in favor of greater de novo photosynthesis during the grain‐filling period, and on the other hand greater relative contribution of the leaf and ear C reserves to remobilization towards the grain. Further splitting the dose increased only the relative contribution of ear C reserves. The stem contribution seemed to be independent of N applied whereas the root contribution tended to diminish with late N application.     

DROUGHT STRESS: Soil Water Availability Alters the Inter‐ and Intra‐Cultivar Competition of Three Spring Wheat Cultivars Bred in Different Eras

Competition for water generates a classic aspect of the tragedy of the commons, the ‘race for fish’, where crops must allocate more resource to acquisition of the limiting resource than is optimal for crop yield allocation. A pot experiment using a simple additive (target–neighbour) design was conducted to examine the above‐ground and below‐ground growth of three spring wheat (Triticum aestivum L.) cultivars when grown alone and in mixtures at three levels of water availability. The effects of competition and water availability were compared by observing patterns of growth, biomass allocation and below‐ground outcomes. Competitive interactions were investigated among cultivars ‘HST’, ‘GY602’ and ‘LC8275’, target plant of each cultivar grown without neighbouring plants are referred to herein as control plant and one target plant of each cultivar sown surrounded either by same or another cultivar as intra‐ or inter‐cultivar competition. Competitive ability was assessed as the response ratio (lnRR) between the target plant surrounded by six other plants and the target plant in isolation. Our results showed that the cultivar ‘HST’, released over a century ago, produced a higher biomass and grain yield than the more recently released cultivars ‘LC8275’ and ‘GY602’ when grown as isolated plants with sufficient water supply. However, competition for resources from neighbours led to target plant biomass and grain yield being significantly reduced relative to controls in all three cultivars, particularly in ‘HST’. When subjected to intra‐cultivar competition, the two recently released cultivars ‘LC8275’ and ‘GY602’ had higher grain yields and water use efficiency for grain than ‘HST’ in all three water regimes. The landrace ‘HST’ had better and significantly linear relationships between biomass and biomass allocation, root length and specific root length, whereas the recent and modern cultivars had much more water‐related species‐specific changes in root morphology and allocation patterns. These results suggest that crop traits that influence competitive ability, such as biomass allocation to roots and root plasticity in response to drought have changed in modern wheat cultivars because of breeding and selection.     

Screening and Analysis of Proteins Interacting with TaPDK from Physiological Male Sterility Induced by CHA in Wheat

To further research the regulatory network of pyruvate dehydrogenase kinase (designated as TaPDK) in physiological male-sterility (PHYMS) of wheat induced by chemical hybridizing agent (CHA) SQ-1, an anther cDNA library was constructed, and the proteins interacting with TaPDK were screened via yeast two-hybrid technique. Subsequently, a few candidate proteins in nucleotide expression levels were detected by real-time quantitative PCR. Yeast-two hybrid screening was performed by mating yeast strain Y2HGold containing BD-TaPDK bait plasmid with yeast strain Y187 including anther cDNA library plasmid. Diploid yeast cells were plated on synthetic dropout nutrient medium (SD/-Ade/-His/-Leu/-Trp) (QDO), and further were incubated on QDO medium containing AbA and X-α-Gal. The interactions between TaPDK and the proteins obtained from positive colonies were further confirmed by co-transformation validation. After plasmids DNA were extracted from blue colonies and sequenced, the sequences results were analyzed by bioinformatic methods. Finally, 24 colonies were obtained, including eight genes, namely non-specific lipid-transfer protein precursor (TanLTP), polyubiquitin (TaPUbi), glyceraldehyde-3-phosphate dehydrogenase, proliferating cell nuclear antigen (TaPCNA), CBS domain containing protein (TaCBS), actin, guanine nucleotide-binding protein beta subunit, chalcone synthase, and three new genes with unknown function. The results of quantitative RT-PCR showed that the expression levels of TanLTP, TaPUbi, and TaPCNA were obviously up-regulated in PHYMS anther, and TaCBS expression was only increased at the tricellular stage in PHYMS anther compared with in fertile lines. Whereas, the expression of TaPDK was obviously down-regulated in PHYMS lines. Collectively, these datas indicated that the majority of candidate proteins might be related to pollen abortion in PHYMS lines, which further suggested that TaPDK plays multiple roles in pollen development, besides participating in regulating pyruvate dehydrogenase complex activity.     

Assessment of the profile of powdery mildew and its damage function at low disease intensities in field experiments with winter wheat

Damage by mildew to winter wheat was studied in 11 field experiments in the Netherlands. Damage is described by the simple function: –0.013 (SE=0.003) kg are^–1 per pustule-day of mildew per leaf, from second node stage to early dough at yield levels of 70 to 90 kg are^–1, in diseasefree plots. No deviations from linearity at disease stresses from zero to two thousand pustuledays per leaf were observed. Years, cultivars or soil types did not affect the damage function significantly. The effects of mildew on some yield components were suggested.Mildew profiles in untreated plots could be described by the equation: CM=CLA^b, in which CM and CLA are the cumulative pustule number and the cumulative leaf area, respectively, calculated both from top to bottom of the canopy, and totals standardized at unity. Estimates of the gradient parameter b aveaged 3.4 (SE=0.9). Observed differences in steepness of the profiles did not affect the damage function significantly.Samenvatting Opbrengstderving van wintertarwe door meeldauw werd bestudeerd in 11 veldproeven in Nederland. De schade bedroeg gemiddeld 0.013 (SE=0.003) kg are^–1 per puistdag meeldauw per blad, vanaf het tweede-knoop stadium tot begin deegrijp bij opbrengstniveaus van 70 tot 90 kg are^–1, in de blanco. Bij een ziektestress van 0 tot 2000 puistdagen meeldauw per blad werd geen afwijking van een rechtlijnig verband gevonden. De schaderelatie werd niet significant beïnvloed door de verschillende jaren, rassen of grondsoorten. Het effect van meeldauw op enkele opbrengstcomponenten werd aangetoond.Meeldauwprofielen in de onbehandelde veldjes konden worden beschreven met de vergelijking: CM=CLA^b, waarin CM het cumulatieve aantal puistjes is en CLA het cumulatieve bladoppervlak, beide berekend van bovenin het bladerdek naar beneden, de totalen gestandaardiseerd op één. De gradiënt parameter b bedroeg gemiddeld 3.4 (SE=0.9). Waargenomen verschillen in steilte van de meeldauwprofielen beïnvloedde de schaderelatie niet aantoonbaar.     

Surveys of cereal diseases and pests in the Netherlands. 2. Stem-base diseases of winter wheat

In the period 1974–1986, ca. 100 commercial winter wheat fields were surveyed annually for stembase diseases. In these years, on average 6% of the tillers was infected with eyespot in spring at the first-second, node stage. Eyespot intensity in spring was high in years with high temperature during winter. Eyespot, intensity in the milky-ripe stage in July, averaged 13% of the culms and was high in years with high temperature in April and high precipitation in March, April and May. These data may improve forecasts.In the milky-ripe stage, 2% of the culms were infected with sharp eyespot, but its intensity has gradually increased during the survey years. No significant correlation of sharp eyespot intensity with dry periods in autumn, spring or summer was found.Symptoms of take-all were found on 0.2% of the stem-bases during ripening. Its intensity was low in years with high precipitation in March, April, May and June and high temperature in May and June.Samenvatting Een honderdtal percelen wintertarwe werd in 1974–86 jaarlijks op voetziekten geïnventariseerd. In het voorjaar, tijdens het, eerste en tweede knoopstadium van het gewas, was gemiddeld 6% van de spruiten aangetast door oogvlekkenziekte. De intensiteit van de ziekte was hoog in jaren met een hoge gemiddelde wintertemperatuur. Tijdens het melkrijpe stadium, in juli was gemiddeld 13% van de halmen aangetast door oogvlekkenziekte. De aantasting in juli was hoog in jaren met een hoge temperatuur in april en veel neerslag in de maanden maart, april en mei. Met deze gegevens kunnen adviessystemen worden verbeterd.Scherpe oogvlekkenziekte was op gemiddeld 2% van de halmen in juli aanwezig. De ziekte nam geleidelijk met de jaren toe. De jaarlijkse intensiteit was niet gecorreleerd met droge perioden in de herfst, voorjaar of zomer.Symptomen van halmdoder waren op gemiddeld 0.2% van de halmen aanwezig. De intensiteit van de ziekte was hoog in jaren met weinig neerslag in maart, april, mei en juni en met een lage temperatuur in mei en juni.     

An advisory model for control of Puccinia recondita in winter wheat

Epidemics ofPuccinia recondita and resulting yield loss of winter wheat were studied in field experiments over three seasons in the Netherlands. Results are reported and used to construct an advisory model for control of brown rust, based on rust monitoring. If the fraction of leaves with rust (I) at a certain development stage is determined, the average number of rust sori per leaf (M) was estimated by: M=EXP(1.84+1.39ln(ln[1/(1-I)]). The final number of sori per leaf (Mf) at early dough was forecast by an exponential growth: Mf=M·EXP(RGR·t). The relative growth rate (RGR) averaged 0.163/day and the forecasting period, t, until early dough, was derived from published data. The forecast number of sori-days per leaf (S, AUDPC-value) was then obtained by: S=(Mf-M)/RGR. Yield loss (kg/ha) by brown rust was 1.15 times the number of sori-days per leaf at low rust intensities. The efficacy of the fungicides used was 85%. The forecast avoidable yield loss (L, kg/ha) was calculated by: L=0.85 (1.15 S). Economic thresholds for brown rust control at different development stages are given for Dutch wheat fields at a cost level of 270 kg/ha for one fungicide application.     

Competition and interactions among stripe rust pathotypes in wheat-cultivar mixtures

A series of experiments was conducted with wheat stripe rust to analyse competition between simple and complex pathotypes in host mixtures. Two different pathotype combinations were tested, with different host components. Each combination included a complex (able to infect two host components) and two simple pathotypes. For one of the combinations, induced resistance was tested in a separate experiment as a possible interaction among pathotypes. Disease severity and pathotype frequencies were measured three times during the epidemic, on each host component grown in pure stands and in mixtures. In one of the experiments, pathotype frequencies were also measured within secondary foci. One of the complex pathotypes appeared to have a low fitness on one of the host components and did not significantly increase in frequency in host mixtures relative to pure stands. The average frequency of the other complex pathotype increased during the first epidemic cycles, but remained stable afterwards, below expected values. The results suggest that the development of complex pathotypes in host mixtures may be influenced by differential aggressiveness on the host components, by induced resistance and by random effects resulting from the formation of disease foci, and depends on pathogen autoinfection rate and dispersal mechanisms.     

Surveys of cereal diseases and pests in the Netherlands. 4. Occurrence of powdery mildew and rusts in winter wheat

Results of annual surveys of winter wheat fields from 1974 to 1986 were compiled to describe epidemics of powdery mildew and rusts in relation to weather and cultivar resistance.An average of 29 and 70% of fields were infected by powdery mildew in May and July, respectively. Mildew prevalence in May was positively correlated with average temperature in October and with average temperature over the months December, January, February and March. In addition, it was correlated negatively with the average grade of mildew resistance of the cultivars sown each year. Prevalence of mildew in July did not show consistent correlations with weather characteristics nor with mildew prevalence in May.Yellow rust was usually not detected in May and on average 18% of the fields was infected in July. The occurrence of yellow rust decreased after 1977, when the farmers adopted cultivars resistant or moderately resistant to yellow rust.Brown rust was usually not detected in May, while in July on average 48% of the fields was infected. Brown rust intensity in July was high in years with a high March temperature and high precipitation during April and May.Black rust was rare in the Netherlands, with 3 and 1% of the fields infected in July 1977 and 1981, respectively.