麦类作物DNA原位杂交及其在远缘杂种染色体分析中的应用

 应用生物素标记的物种基因组DNA探针和克隆的专化性探针，对六倍体小黑麦双二倍体及其杂种和八倍体小麦簇毛麦双二倍体及其杂种进行了染色体DNA原位杂交。结果表明，应用这两种探针均可以准确地检测出小麦远缘杂种中的异源染色体和染色体片段。先后对二个六倍体小黑麦双二倍体（Badger、M#-2A）、一个八倍体小簇麦双二倍体、三个小黑麦易位系（宁麦8026、Amigo、Apollo）、一个小黑麦代换系（84056）和二个小簇麦附加系（94009-5-4、94009-5-9）进行原位杂交和异源染色体的检测，均获得较好的结果。此外，还讨论了应用生物素标记的探针进行原位杂交可能出现的一些技术问题。     

青蒿素对小麦种子萌发及幼苗生长的影响

为了解青蒿素对小麦种子萌发和幼苗生长的影响,运用室内培养皿法,分析了不同浓度的青蒿素溶液浸种后小麦种子发芽率及幼苗高度、根长、整株鲜重、根和茎鲜重及干重、根系活力等指标的变化。结果表明,青蒿素对小麦种子萌发及幼苗生长有抑制作用,且浓度越高,抑制作用越显著。与无菌水对照相比,青蒿素浸种处理显著降低了小麦的种子发芽率、苗高、根长、根鲜重和茎的鲜、干重,增加了根干重和根冠比。小麦的根系活力和可溶性蛋白含量随青蒿素浓度的增加而递减,可溶性糖含量则随之递增,叶绿素含量则呈先升高后降低的趋势,以20mg·L-1处理含量最高。此外,青蒿素浸种处理对小麦幼苗各项指标的影响与15mg·L-1多效唑相似。     

Effect of mannitol pretreatment to improve green plant regeneration on isolated microspore culture in Triticum turgidum ssp. durum cv. 'Jennah Khetifa'

The use of doubled haploids improves the efficiency of cultivar development in many crops and can be helpful in genetic and molecular studies. The major problem with this approach is the low efficiency of green plant regeneration. We describe here an efficient method for inducing embryos and regenerating green plants directly from isolated microspores of durum wheat cv. ‘Jennah Khetifa’. Tillers from donor plants were pretreated in 0.3 m mannitol and were stored at 4°C at various times: 3, 5, 6, 7, 8, 10 and 12 days. Our results showed clearly that the novel pretreatment combined mannitol 0.3 m and cold for 7 days had a strong effect on the number of embryos produced and regenerated green plants. Under this condition 13 475 mature embryos were produced from 2 693 500 microspores. Moreover, 85 green plants were obtained. High green plants regeneration frequency was recorded. As an average 11.55 green plants were produced per 100 000 microspores (about the equivalent of six plants per spike). Therefore, this study showed clearly that our results are the best ones published until now in durum wheat.     

小麦籽粒淀粉分支酶同工酶基因型与酶活性关系研究

测定了71个小麦品种的籽粒淀粉分支酶活性,并采用非变性聚丙烯酰胺凝胶电泳（Native-PAGE）,鉴定了分支酶同工酶的基因型,以明确小麦籽粒淀粉分支酶（SBE）各同工型等位基因的分布特点和基因型组成,阐明分支酶活性与分支酶同工酶基因型的相关性。结果表明,分支酶有SBEⅠ和SBEⅡ两个基因位点,SBEⅡ位点未显现多态性,SBEⅠ位点存在多样性。在SBEⅠ位点鉴定出A、B、D_ⅰ和D_ⅱ共4个等位基因位点,各等位基因位点出现频率分别为9.9%、76.1%、95.8%和57.8%;共检测到9种基因型,其中基因型D_ⅰD_ⅱB、D_ⅰB和D_ⅰD_ⅱ有较高的分布频率,分别为45.1%、28.2%和8.5%。从单一位点分析,分支酶活性与SBEⅠ位点有显著的相关性,与SBEⅡ位点相关性不显著。由此可见,小麦籽粒胚乳淀粉分支酶同工酶不同基因位点和基因型对酶活性具有不同的遗传效应。     

Relationship between allelic variation of Glu-1 and Gli-1/Glu-3 prolamin loci and gluten strength in hexaploid wheat

Summary Allelic variation of prolamin loci was examined in the F₂ from crosses between the hexaploid wheat varieties: Cajeme 71, Yécora 70, Ablaca, Anza, Pané 247 and Axona. Different allelic blocks for gliadins and LMW glutenin subunits were determined in Gli-1, Gli-2 and Glu-3 loci. A percentage of recombination of 1.5 ± 0.3 was determined between Gli-A1 and Glu-3 in the F₂ progeny of Yécora 70 x Axona. A significant positive association was found between gluten strength, measured by SDS-sedimentation volume, and the prolamins coded by Anza Gli-D1/Glu-D3 loci and Yécora 70 Gli-A1/Glu-A3 loci. Interactions between non homeologous loci Glu-1 and Gli-1/Glu-3 were also found.     

Evaluation of common wheat cultivars for tan spot resistance and chromosomal location of a resistance gene in the cultivar 'Salamouni'

A total of 50 wheat (Triticum aestivum L.) cultivars were evaluated for resistance to tan spot, using Pyrenophora tritici‐repentis race 1 and race 5 isolates. The cultivars ‘Salamouni’, ‘Red Chief’, ‘Dashen’, ‘Empire’ and ‘Armada’ were resistant to isolate ASC1a (race 1), whereas 76% of the cultivars were susceptible. Chi‐squared analysis of the F₂ segregation data of hybrids between 20 monosomic lines of the wheat cultivar ‘Chinese Spring’ and the resistant cultivar ‘Salamouni’ revealed that tan spot resistance in ‘Salamouni’ was controlled by a single recessive gene located on chromosome 3A. This gene is designated tsn4. The resistant cultivars identified in this study are recommended for use in breeding programmes to improve tan spot resistance in common wheat.     

Relationship between Carbon Isotope Discrimination, Ash Content and Grain Yield in Wheat in the Peninsular Zone of India

Carbon isotope discrimination (Δ) and ash content (m_a) have been proposed as indirect selection criteria for grain yield in wheat. The associations between Δ, m_a and grain yield were found, however, to depend highly on the environmental conditions, the organ sampled and the time of sampling. In this study, carried out in the warm conditions of the Peninsular Zone of India, the relationship between Δ, m_a and yield was studied in 30 bread and durum wheat cultivars under residual soil moisture stress (RSMS), post‐anthesis water stress (PAWS) and well‐watered (WW) conditions. Both Δ and m_a were analysed in young seedlings (four‐leaf stage), leaves at anthesis and grain at maturity. Ash content was also evaluated in leaves at booting stage and maturity. Grain Δ was lower under PAWS and RSMS than under WW, while seedling and leaf Δ did not significantly differ among water regimes. Grain yield was positively correlated to grain Δ under PAWS and negatively correlated to grain m_a under RSMS. A significant positive correlation was noted under RSMS and WW treatments between m_aL_m and grain yield. Ash content in leaf at maturity consequently appears to be a useful indirect selection criterion in environments where Δ does not show any correlation with yield. The results highlight the potential of Δ and m_a as indirect selection criteria for wheat yield in the conditions of the Peninsular Zone of India.     

Assessment of partial resistance to powdery mildew in Chinese wheat varieties

Field trials in two cropping seasons and two locations in central China were conducted on 60 Chinese autumn‐sown wheat varieties to assess their partial resistance to powdery mildew. Mean levels of disease severity ranged from close to 0 to more than 90%. The method of inoculation and the location in which trials were conducted affected the relative performance of the varieties, but these effects were much smaller than the main effect of variety. The area under the disease progress curve was highly correlated with final disease severity, but both were poorly correlated with apparent infection rate. Disease severity was regressed against frequencies of virulence in the Blumeria graminis (syn. Erysiphe graminis) f sp. tritici populations in the trial plots. A vertical distance (D) from the mean mildew severity to the fitted line was calculated for each variety and was used to quantify partial resistance. Five of the 60 varieties, ‘Hx8541’, ‘E28547’, ‘Chuan1066’, ‘Zhe88pin6’ and ‘Lin5064’, consistently expressed relatively low levels of disease despite high frequencies of virulence in the pathogen and had consistently high D‐values. They may therefore have good levels of partial resistance.     

In vitro Selection for Tolerance to Fusarium in F1 Microspore Populations of Wheat

Microspore populations of eight Fhybrids of winter wheat (Triticum aestivum L.) whose parents had different levels of resistance to Fusarium were screened in vitro, using phytotoxins of Fusarium as biochemical probe. Two selection methods were compared for the in vitro selection: either embryoids and calli were first initiated from anthers in toxin-free medium and then grown on medium with 0.3—0.9 %Fusarium toxin; or anthers were immediately cultured in modified liquid potato-2 medium in the presence of 1.5, 2.0 and 2.5ml Fusarium toxin per liter culture medium, a concentration which reduced the number of calli and embryoids to about 10 % compared to the toxin-free controls. Microspores from donor hybrids which were produced from very susceptible cultivars were killed by lower toxin concentrations than micro-spores from hybrids of less susceptible parents. From surviving calli and embryoids, originally initiated from 242,000 anthers in both procedures a total of 375 green lines could be regenerated. The results indicate that it is possible to enrich the fraction of regenerating microspores by those which contain the gene complex responsible for reduced susceptibility to Fusarium by the use of a pathotoxin.     

Rapid Screening for Aluminum Tolerance in Cereals by Use of the Chlorophyll Fluorescence Test

The effects of aluminum on the photosynthetic apparatus were examined in cereals grown in nutrient solutions (pH 4.5) at two Al levels (0 and 148 μM). The chlorophyll fluorescence kinetics results confirmed that the soft wheat ‘BHG’ cultivar has the potential for growth on acid soils while triticale cultivars ‘Niovi’ and ‘Dada’ appeared to be relatively tolerant. The percentage decrease in F_v/F_m of the less tolerant cultivars after Al-treatment indicated a decrease in the efficiency of the primary photochemistry of PS II, while the decrease in the ratio F_V/F_osuggested that exposure of the cultivars ‘Dio’ and ‘Appulo E’ to aluminum caused injury to the thylakoid structure. The percentage fluctuations of the ratio F_v/F_mwere shown to correlate very closely with the assessment of injury as evaluated by the relative top fresh weight. Measurements of chlorophyll fluorescence in vivo could be used to monitor injury caused by “Al-stress”, and thus they may serve as a rapid screening test for Al tolerance.