A rapid,automated method for measuring α-amylase in pre-harvest sprouted (sprout damaged) wheat

The quality of wheat for baking is critically dependent on the level of α-amylase (1,4-α-D-glucan glucanohydrolase, EC 3.2.1.1), which can be present as “late maturity α-amylase” (LMA), or due to pre-harvest sprouting due to high rainfall and humidity at the time of harvesting. The most commonly used method to measure α-amylase in wheat grain is the Hagberg Falling Number method, but values are also influenced by rheological properties of starch in the grain. In this study we describe a simple, rapid, automated method (Amylase SD) for measurement of α-amylase in pre-harvest sprouted (sprout damaged) wheat grain. The method (Amylase SD) measures the release of p-nitrophenol from 4,6-O-ethylidene-α-4-nitrophenyl-maltoheptaoside by α-amylase in the presence of α-glucosidase. The absorbance of p-nitrophenolate measured at 405 nm in a ChemWell^®-T auto-analyser is directly related to the level of α-amylase activity present in the milled wheat grain extract. The Amylase SD method generated <6%CV and correlation to the Falling Number method was represented by an inflection point at ∼160 s. The precision, sensitivity and speed of this method provides an ideal alternative to the Falling Number method for measurement of α-amylase (sprout damage) in wheat grain in wheat breeding programmes or at grain receival points.     

The use of <Emphasis Type="Italic">Vp1</Emphasis> in real time RT-PCR to select for pre-harvest sprouting tolerance in triticale

In spite of the availability of laboratory and field tests there is still a major problem to select pre-harvest sprouting (PHS) tolerant triticale varieties in a reliable, field-independent way. One approach to minimize the influence of environmental conditions and physio-morphological traits on PHS detection is using molecular genetic tools. The ‘viviparous’ Vp1 gene has been repeatedly described to play an important role in dormancy in wheat. A quantitative RT-PCR assay based on the expression of the Vp1 gene has been developed. Specific primers were designed for detecting Vp1 in both wheat and triticale. The expression levels of Vp1 were normalized using reference genes and relatively quantified with the comparative C_t-method. However, the first results indicate that the achieved Vp1 expression levels at 50 days post anthesis are not useful to select for PHS tolerance, both in wheat and triticale. This negative outcome so far is possibly due to the existence of several splicing events or to the late assaying moment in the kernel development, when Vp1 expression is found to be low.     

RL4137 contributes preharvest sprouting resistance to Canadian wheats

Preharvest sprouting (PHS) in spring wheat (Triticum aestivum L.) causes significant economic losses due to a reduction in grain functionality, grain yield and viability of seed for planting. Genetic resistance to PHS reduces these losses. Development of PHS resistant cultivars is complicated by the effects of genotype, environment, kernel diseases and spike morphological factors. RL4137 has consistently exhibited high levels of resistance to PHS over years and environments. The mean PHS scores of Canada Western Red Spring (CWRS) wheat cultivars with RL4137 in their ancestry are lower than that of CWRS wheat cultivars without. RL4137 has two mechanisms for PHS resistance, one associated with kernel color and the other not associated with kernel color. RL4137 was the source of PHS resistance in white wheats HY361, AC Vista, Snowbird, Kanata, and Snowstar, all of which had significantly lower PHS scores than the white-seeded check, Genesis. Known DNA markers relating to PHS were used to compare haplotypes with and without RL4137 in the ancestry. Coefficients of parentage also demonstrated the relationship. Because cultivars that have RL4137 in their ancestry were grown on about 77% of the spring wheat area for 2003–2007, RL4137 continues to contribute to protecting market grade from preharvest sprouting.     

Grain dormancy in fixed lines of white-grained wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.) grown under controlled environmental conditions

Pre-harvest sprouting (PHS) in wheat (Triticum aestivum L.) can be a significant problem, causing deleterious effects on grain quality. However, the adverse impacts of PHS can be reduced by introgressing genes controlling grain dormancy into white-grained bread wheat. Screening for grain dormancy typically involves germination testing of harvest-ripe grain grown in a glasshouse or field. However, the more uniform environmental conditions provided by temperature controlled glasshouses (i.e. controlled environmental conditions—CEC) may provide significant benefits for the assessment of grain dormancy. In this study, the dormancy phenotype of grain grown under CEC incorporating an extended photoperiod, was compared with 2 years of data from field grown material. Four dormant double haploid lines (derived from SW95-50213 and AUS1408) and two locally adapted non-dormant cultivars EGA Gregory and EGA Wills were compared in three replicated experiments grown under CEC (22 ± 3°C and 24 h photoperiod). The germination response of harvest-ripe grain was examined to assess the expression of grain dormancy. Two measures of germination, the predicted time to 50% germination (G ₅₀) and a weighted germination index, both clearly differentiated dormant and non-dormant lines grown under CEC. In addition, levels of grain dormancy were similar to field-grown plants. These results demonstrated that CEC with an extended photoperiod can be used for rapid and reliable characterisation of grain dormancy in fixed lines of bread wheat.     

Patterns of variation in potato dihaploid families

Summary Most dihaploids derived from cv. Pentland Crown had higher yields, when expressed as percentages of their parent's yield, than did those of cv. Record. Some genes of Pentland Crown may have a greater additive effect on yield than those of Record but there may also be differences in deleterious recessives which are expressed in their dihaploids. The frequency distributions of specific gravities and sprout lengths of the dihaploids were similar for both cultivars. Some genes may have an additive effect on specific gravity but the small sprout lengths of dihaploids may be due to their often weak constitution caused by the expression of deleterious genes. The distribution of foliage blight (Phytophthora infestans) resistance in dihaploids of resistant tetraploids indicates that multiplex genes may determine the resistances in the parents.

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Zusammenfassung Ertrag, mittleres Knollengewicht und die Knollenzahl pro Pflanze von Dihaploiden der Sorten Pentland Crown und Record (Abb. 1,2 und 3) zeigen, dass sich Unterschiede (signifikant für Ertrag und mittleres Knollengewicht,P<0,01) zwischen beiden Sorten ergeben k?nnen, wenn der Ertrag und seine Komponenten genetisch determiniert waren. Die meisten Dihaploiden von Record hatten weniger als 10% der Ertr?ge ihrer Eltern, w?hrend die Ertr?ge der meisten Dihaploiden von Pentland Crown zwischen 20 und 50% schwankten. Ein betr?chtlicher Teil von Pentland Crown-Dihaploiden hatten h?here als modale Werte für mittleres Knollengewicht und mittlere Knollenzahl pro Pflanze. Mehrere Gene von Pentland Crown und ihrer Dihaploiden dürften deshalb additive Einflüsse auf den Ertrag und seine Komponenten haben. Deshalb dürfte von diesen Klonen eine signifikante generelle Kombinationsf?higkeit (GCA) für diese Eigenschaften erwartet werden, wenn sie bei der Züchtung verwendet werden. Erste Daten von tetraploiden Nachkommen von dihaploid × tetraploid-Kreuzungen (siehe Tabelle I mit der Herkunft des Materials) zeigen jedoch, dass solche GCA-Effekte nicht wesentlich waren. Dies k?nnte so sein, weil die spezifische Kombinationsf?higkeit wichtiger war oder weil Unterschiede in Zahl und St?rke von nachteiligen Genen, die vorher bei den Sorten verborgen waren, vorhanden waren. Tabelle 2 zeigt die Korrelationskoeffizienten zwischen Ertrag, mittlerer Knollenzahl, mittlerem Knollengewicht, spezifischem Gewicht und Keiml?nge für Dihaploide von Pentland Crown und Record. Dabei ergaben sich signifikante Korrelationen zwischen Ertrag und seinen Komponenten, aber keine signifikante negative Korrelation zwischen Knollenzahl und mittleren Knollengewicht. Einige Dihaploide hatten ein h?heres spezifisches Gewicht als ihre tetraploiden Eltern. Die spezifischen Gewichte der Dihaploiden waren um ein Mittel verteilt, welches ann?hernd gleich mit dem der Eltern beider Sorten (Abb. 4) war; vermutlich waren sowohl additive als auch nicht-additive Effekte für die Determinierung des spezifischen Gewichtes von beiden Eltern erforderlich. Die Keiml?ngen dihaploider Knollen waren durchweg gering (Abb. 5), dies dürfte auf die Anwesenheit nachteiliger rezessiver Gene zurückzuführen sein, welche die oft nur schwachen Konstitutionen von Dihaploiden beider Sorten kennzeichnen. Genetische Unterschiede zwischen Sorten, welche anderenfalls als unterschiedliche dihaploide Variationsmuster bezeichnet worden w?ren. k?nnten deshalb maskiert gewesen sein. Hohe Grade an quantitativer Resistenz gegenüber Krautf?ule (Phytophthora infestans) in den meisten Dihaploiden von zwei resistenten Tetraploiden zeigten, dass einige dominante Gene, die die Resistenz bestimmen, multipel sein k?nnten (Abb. 6). Zus?tzlich zur Informationssteigerung über die genetische Determinierung landwirtschaftlich wichtiger Merkmale in Tetraploiden ist die Produktion von Dihaploiden eine Methode zur ?nderung von Gen-Verbindungen ohne Einführung von fremden genetischem Material. Es k?nnten mehr erwünschte Gen-Kombinationen ermittelt werden, die als Resultat für Kartoffel-Züchtungsprogramme ausgebeutet werden k?nnten.

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Résumé Le rendement, le poids moyen de tubercules et le nombre de tubercules par plante des diplo?des de Pentland Crown et Record (fig. 1, 2 et 3) montrent qu'il peut y avoir des différences (significatives pour le rendement et le poids moyen de tubercule,P<0.01) entre les deux variétés, du fait que les composantes du rendement étaient déterminées. La plupart des diplo?des de Record ont des rendements inférieurs de 10% à ceux de leurs parents, alors que les diplo?des de Pentland Crown donnent des rendements compris entre 20 et 50%. Une proportion assez grande de diplo?des de Pentland Crown a un poids moyen de tubercules et un nombre moyen de tubercules par plante plus élevés que les valeurs modales correspondantes. Quelques gênes de Pentland Crown et de diplo?des issus de cette variété peuvent avoir par conséquent un effet positif sur le rendement et ses composantes; ces cl?nes pourraient en définitive permettre des combinaisons générales pour les caractères utilisés en sélection. Des résultats préliminaires sur tétraplo?des obtenus à partir de croisements diplo?des × tétraplo?des (tableau 1) ont montré cependant que de tels effets de combinaisons générales n'étaient pas importants. Ceci est d? peut être du fait que l'aptitude spécifique à la combinaison est plus importante ou qu'il y a eu des différences dans le nombre et l'ampleur des gênes défavorables, masqués au départ dans les variétés. Le tableau 2 donne les coefficients de corrélation entre le rendement, le nombre moyen de tubercules, le poids moyen de tubercules, le poids spécifique et la longueur des germes pour les diplo?des de Pentland Crown et Record. Des corrélations significatives ont été observées entre le rendement et ses composantes mais pas entre le nombre de tubercules et leur poids moyen. Quelques diplo?des ont des poids spécifiques plus élevés que ceux de leurs parents tétraplo?des. Pour chaque variété, les poids spécifiques des diplo?des ont une distribution par rapport à la moyenne à peu près semblable à celle de leurs parents (fig. 4) et les effets génétiques additifs et non-additifs sont probablement importants dans la détermination du poids spécifique pour chaque parent. Les germes des tubercules diplo?des sont généralement courts (fig. 5) et ceci provient sans doute de la présence de gênes récessifs défavorables qui déterminent des constitutions souvent chétives des diplo?des de ces deux variétés. Les différences génétiques entre les variétés peuvent par conséquent être marquées, alors qu'elles auraient été mises en évidence par des exemples de variation entre différents diplo?des. De hauts niveaux de résistance quantitative au mildiou du feuillage (Phytophthora infestans) obtenus avec la plupart des diplo?des provenant de deux tétraplo?des résistants au mildiou indiquent que certains gènes dominants gouvernant la résistance peuvent être multipliés (fig. 6). Afin de mieux conna?tre la détermination génétique des caractères agronomiques importants des tétraplo?des, la production de diplo?des est une méthode d'association de gènes remaniés, sans l'introduction de matériel génétique étranger. Davantage de combinaisons de gènes souhaitables peuvent être identifiées afin d'être exploitées dans un programme de sélection de pommes de terre.

     

ASI蛋白的特性及其应用研究进展

ＡＳＩ是抑制小麦α－淀粉酶活性及改善小麦品质的一种重要蛋白,目前已将编码ＡＳＩ蛋白的基因定位于麦类作物第二同源染色体的长臂上。ＡＳＩ蛋白具有高度的多态性。大麦中ＡＳＩ蛋白对小麦α－淀粉酶的活性抑制作用较强,在小麦中导入大麦ＡＳＩ蛋白基因能够延缓小麦穗发芽。抗ＡＳＩ蛋白的单克隆抗体具有特异性,依赖抗小麦ＡＳＩ蛋白的ＥＬＩＳＡ是测定ＡＳＩ蛋白浓度的一种有效方法。本文对α－淀粉酶抑制蛋白的特性及应用进行     

Epicuticular waxes and regulation of ear wetting and pre-harvest sprouting in barley and wheat

Morphological features of the cereal ear, including awns, alter pre-harvest sprouting damage by changing the rate of water absorption during rainfall. In this paper, the potential for wheat (Triticum sp.) arid barley (Hordeum vulgare L.) waxes to reduce sprouting by increasing water repellency of the mature ear has been examined. Six barley F₂ populations segregating for different non-glaucous single-gene mutants controlling waxes on ears were examined. Water repellency was assessed by measuring both the contact angle of a water drop placed on the lemma surface (internal angle) and by repetitive weighings of whole ears during their exposure to simulated rainfall. The lemma of glaucous (wild type) lines had larger water drop contact angles, an indication of poorer spread of water over the surface. In simulated rainfall, ears of the glaucous lines showed a clear reduction of wetting (20–30% less) and, after 72 h of wetting, their in-ear sprouting was reduced by 50 to 65%. When pre-wet, the glaucous ears also shed water more readily when shaken to simulate the combined effect of wind and rainfall. To reduce pre-harvest sprouting of barley it may be possible to screen visually for ears that are more glaucous but a more specific screen would be to select for lemma water drop contact angle since it is a good indicator of ear wettability and so allows differences in surface properties to be assessed. For bread wheat (T. aestivumL.), as for barley, the more glaucous the ear, the greater the water drop contact angle and the more tubular surface wax coverage seen in scanning electron microscope images. In addition, surface wax amount apparently affected in-ear wettingin lines of durum wheat, (T. turgidum L.). Possible genetic relationships between waxy/waxless genes in wheat and barley are suggested with the aim, ultimately, of altering ear glaucousness to give increased water repellency and a reduction of in-ear sprouting of wheat. This revised version was published online in July 2006 with corrections to the Cover Date.     

H2CN2和CaCN2对酿酒葡萄萌芽开花及成熟期的影响

试验于2001年10月-2002年8月在攀枝花进行。处理药剂及其浓度为16.7?CN2、0.5%~2.0%H2CN2、0.5%~2.0%H2CN2 1%吐温80;处理时期为12月25日、1月10日、1月25日。最佳处理时间是1月10日;最佳处理组合是1.0%H2CN2 1.0%吐温80,霞多丽、佳美、梅尔诺、黑虎香分别提前萌芽16、13、11、4d;提前成熟17、13、14、6d;提高萌芽率4%、12%、17%、9%;芽眼成枝率提高40%、29%、37%、12%;提高果枝率为9%、8%、28%、4%,提高结果系数7%、7%、37%、3%。不同品种对处理反应有差异,对照黑虎香有药害现象。     

杉木干基萌芽能力的遗传变异

分析了两片杉木子代林伐桩和杉木无性系采穗圃萌条数量的遗传变异情况。结果表明：杉木子代林伐桩萌条数量在产地间、产地内家系间及组合间均存在极显著差异，并有中等以上遗传力；遗传变异系数在３７％～３９％之间；产地的遗传方差分量与产地内家系间的相近。伐桩萌条数量与基径粗度的关系随伐桩年龄的变化而表现不一；不同砍伐季节伐桩萌条数量也存在较大差异。杉木无性系采穗圃无性系间有效萌条数量存在极显著差异，无性系的萌芽能力有较高的重复力。     

不同穗发芽抗性的小麦胚对ABA敏感性及抗性机制研究

以20个小麦品种为材料,测定了不同穗发芽抗性的小麦品种处于种子发育过程中的胚对ABA敏感性和后熟过程中种子的萌发能力,以揭示小麦对穗发芽的抗性机制。结果表明,在种子发育过程中,不同抗性品种的胚萌发能力和胚对ABA敏感性存在差异。胚萌发能力随胚龄的增加,呈"低～高～低"的变化趋势。扬花后25d到35d的胚萌发势的降幅值在抗穗发芽基因型中所占的比重明显较易穗发芽基因型大。易穗发芽基因型对ABA的敏感性随胚萌发能力的提高而下降,而抗穗发芽的基因型对ABA的敏感性随胚萌发能力的提高则有升有降。因此,胚休眠主要在发育后期获得。红皮易穗发芽品种的成熟胚因脱离母体而提高的萌发势明显较白皮易穗发芽品种的高。品种的成熟胚离体萌发能力越高,则种子休眠期越短。