白皮小麦收获前穗发芽及品种抗性机制探讨

对４８个白皮小麦种质及２个红皮小麦种质连续３年测定结果,品种间收获前穗发芽率,籽粒发芽率及收获后种子休眠特性差异显著。穗发芽敏感性因年份和种子发育时期而异,开花后３５－４０天较敏感,品种间差异大。休眠期长短与穗发芽率间呈极显著负相关。收获后的种子α－淀粉酶活性,降落值,与穗发芽率间分别呈极显著正相关和极显著负相关。     

Novel resistance to pre-harvest sprouting in Australian wheat from the wild relative Triticum tauschii

The diploid D-genome progenitor of hexaploid wheat, Triticum tauschii (Coss.) Schmahl., was screened to identify mechanisms for resistance to pre-harvest sprouting. A number of promising mechanisms were identified, and transferred to hexaploid wheat via wide-hybridisation. One identified mechanism, an inhibitory phenolic compound present in the bracts surrounding the grain, has been shown to function effectively in synthetic hexaploid wheats. A number of seed-borne dormancy mechanisms were also identified. Expression of embryo dormancy in synthetic hexaploid wheats was demonstrated when compared with non-dormant hexaploid wheat. Effects of the seed coat on dormancy were also studied, with the seed coat of synthetic hexaploids accelerating rather than inhibiting germination. Embryo dormancy was also demonstrated in two `direct-cross' hybrids. The results suggest that a combination of the described mechanisms may produce white wheats with resistance to pre-harvest sprouting adequate for most Australian climatic conditions. This revised version was published online in August 2006 with corrections to the Cover Date.     

Dormancy breaking procedures and the breeding of white-grained wheat with resistance to pre-harvest sprouting

Summary Gibberellic acid, cold (4°C) and a combination of these two treatments were tested for use in breaking dormancy in 27 lines of white-grained wheat with varying levels of resistance to pre-harvest sprouting. Germination increased in all lines treated with gibberellic acid. Dormancy could be broken with 1 M gibberellic acid. Response to cold varied. A combination of gibberellic acid and cold treatment was the most effective. This technique has been found useful in treating seed in a breeding program aimed at producing wheats with resistance to pre-harvest sprouting.     

Seed development-related expression of ARGONAUTE4_9 class of genes in barley: possible role in seed dormancy

Resistance to pre-harvest sprouting is an important breeding objective for cereal crops like barley and wheat. Seed dormancy, which determines the resistance or susceptibility to pre-harvest sprouting (PHS), is a complex trait. It is largely controlled by the antagonistic action of the plant hormones abscisic acid and gibberellic acid, but also has a large component of genotype?×?environment interaction. Recent studies have revealed a role for epigenetic changes through histone modification in controlling seed dormancy. However, the role of DNA methylation in seed development and dormancy is not known. In this study, we explored the role of ARGONAUTE4_9 class genes of the DNA methylation pathway in seed development and dormancy in barley. Our results show that the two AGO4_9 class genes in barley, i.e. AGO1002 and AGO1003, are preferentially expressed in ovaries at meiosis and in embryos 25?days after pollination (DAP). The expression of AGO1003 is two to fivefold higher than that of AGO1002 in these tissues, demonstrating differential expression of these genes. We also analysed the expression of AGO1003 in embryos of PHS-resistant and -susceptible varieties at 25?DAP and found a significant variation in the expression of this gene in seeds of dormant and non-dormant lines. The observed expression pattern of AGO1002 and AGO1003 suggests a possible role in sporogenesis and post-fertilization seed development. Indirectly these results imply a potential role of DNA methylation in seed development and seed dormancy.     

小麦籽粒休眠Vp1-B1基因的等位变异检测与分离

Vp1基因是控制籽粒休眠性的重要基因之一,检测该基因的等位变异类型,可以进一步理解籽粒休眠以及穗发芽抗性的遗传控制机理.本文根据GenBank中小麦Vp1-B1基因序列设计特异引物检测我国小麦微核心种质以及地方品种和推广品种,结果表明,本研究除了发现已报道的3种等位变异类型,分别为Vp1-B1a、Vp1-B1b和Vp1-B1c,另外检测到一种新的变异类型,暂命名为Vp1-B1x.经改良的变性PAGE凝胶电泳检测以及序列比对分析表明,Vp1-B1x与Vp1-B1c基因的序列相似性最高,达99%;其在第3内含子区域发生"ATAT"4个碱基的插入以及4个SNP,但是该等位类型在所检测的品种中分布较少,其与籽粒休眠水平及穗发芽抗性之间的关系尚待进一步验证.     

四川省小麦品种(系)穗发芽抗性评价

穗发芽严重影响小麦产量和品质,受到生长环境、种皮颜色和遗传基因的共同控制。由于四川地区在小麦收获季节容易出现高温高湿的气候条件,是小麦穗发芽的常发和重发区。依据材料的穗发芽抗性,有目的性地配制杂交组合选育抗性品种是控制小麦穗发芽的重要策略,而系统鉴定分析各优良品种及品系的穗发芽抗性是亲本选配的重要依据。本研究调查分析了242份2000年来四川省审定小麦品种及即将成为未来几年新品种的2017—2018 年四川省区试品系的田间穗发芽情况,发现分别有23(9.5%)和67 (27.7%)份材料具有强（穗发芽率低于5%）和高（穗发芽率介于5%~15%）的穗发芽抗性。这为穗发芽抗性育种亲本的选择提供了参考;并且发现穗发芽抗性在不同类型材料中存在差异：2000 年来审定品种＜区试普通小麦品系＜区试早播早熟小麦品系＜区试特殊用途小麦品系,穗发芽抗性在未来几年品种中将有所增强;种皮颜色对穗发芽抗性具有强烈影响,随着种皮颜色加深,穗发芽抗性越强,但仍发现3 份（‘川育26’、‘川辐9 号’、‘川辐17 号’）白皮小麦材料具有强的穗发芽抗性,为白皮小麦抗穗发芽育种提供参考。其次发现在四川环境下,早播可能会提高小麦品种的穗发芽抗性。     

Developing standardized methods for breeding preharvest sprouting resistant wheat, challenges and successes in Canadian wheat

Preharvest sprouting (PHS) in spring wheat (Triticum aestivum L.) and durum wheat (T. turgidum L. var durum) causes significant economic losses due to a reduction in grain yield, grain functionality and viability of seed for planting. Average annual estimated losses in Canada are about $100 million. Genetic resistance to PHS reduces these losses. Development of PHS resistant cultivars is complicated by the effects of factors under genetic control, such as spike morphology, seed dormancy, environment, and kernel diseases. Resistance to PHS has been a breeding priority since the late 1960s. Development of RL4137, which is the primary source of PHS resistance in the Canada Western Red Spring market class, has led to cultivar improvements. A white-seeded derivative of RL4137 is the primary source of PHS in the Canada Prairie Spring White and Canada Western Hard White Spring wheat market classes. Procedures to select for PHS resistance vary among breeding programs, market classes and by degree of inbreeding. Methods include artificial sprouting of intact spikes, germination tests, natural weathering in field trials, artificial weathering trials, and indirect assessment of sprouting by measuring Hagberg falling number. Although many genetic loci have been attributed to preharvest sprouting resistance, application of molecular markers is currently limited due to the complex inheritance of the trait. In Canada, cultivars are characterized for their relative level of PHS resistance and the information is made available to producers.     

Geographical differentiation and diallel analysis of seed dormancy in barley

Seed dormancy is one of the most important parameters affecting the malting process and pre-harvest sprouting in barley (Hordeum vulgare L.). Variation of seed dormancy in 4365 cultivated and 177 wild barley (ssp. spontaneum) accessions derived from different regions of the world was investigated in Okayama University, Kurashiki, Japan. Seed dormancy of each accession was estimated from their germination percentages at 0, 5, 10 and 15 weeks post-harvest after-ripening periods. All of the wild barley accessions showed less than 10% germination at 0 week after-ripening period. Level of seed dormancy in 4365 cultivated barley accessions showed a clear geographical differentiation. Seventy seven percent of Ethiopian accessions showed high germination percentages, while 86% of Japanese, Turkish and North African accessions showed low germination percentages at 0 week after-ripening period. A half diallel cross using eleven barley accessions with different level of dormancy revealed that seed dormancy was predominately controlled by additive gene effects. These results suggest that large genetic diversity for seed dormancy in barley is explained as different levels of additive accumulation of genetic factors. Barley varieties showing appropriate dormancy could be developed by crossing among barley germplasm accessions used in the present study.     

On the problem of pre-harvest sprouting of wheat

Summary The problem of pre-harvest sprouting in wheat is very intricate. New concepts in breeding for resistance to pre-harvest sprouting regard many components such as -amylase production potential, response to gibberellic acid and inhibition in the bracts as important as dormancy. In the research work carried out at The University of Sydney's Plant Breeding Institute, we found varietal differences for each of these components. We also developed criteria for screening breeding material for these components. The implications of our findings in planning efficient breeding programmes to evolve varieties with multiple resistance to pre-harvest sprouting have been discussed briefly.     

Predicting pre-harvest sprouting susceptibility in New Zealand wheat cultivars

New Zealand has a maritime climate, hence conditions conducive to pre-harvest sprouting (PHS) in the cereal-growing regions are common. Wheat growers are faced with an ever-increasing list of potential cultivars, and the risk of sprouting damage is a very important consideration in choosing among them. Supported by the NZ Foundation for Arable Research and Flour Millers Association, sprouting nurseries based on artificial weathering and seed dormancy testing have been operated for the last three seasons in an attempt to quantify the risk level of sprouting for each cultivar. In late January/early February 2004, much of the New Zealand wheat crop, including five cultivar evaluation trial sites, was exposed to conditions conducive to pre-harvest sprouting. Data from a sample of the cultivars tested in both the 2003 and 2004 sprouting nurseries and these trials were analysed to assess the repeatability of the various tests used and their usefulness for predicting downgrading due to rainfall.     

Temperature effects on seed germination and expression of seed dormancy in wheat

Because preharvest sprouting decreases quantity and quality of wheat grain, researchers need effective protocols to assess response to preharvest sprouting conditions. The aim of this study was to determine which temperature gives the greatest difference in seed germination and expression of seed dormancy in 10 spring wheat genotypes. The genotypes were grown in the field near Swift Current, Saskatchewan in 2000 in a randomized complete block with four replicates. Seed samples were harvested at approximately 25% moisture content (wet weight basis) and dried to 12% moisture content with minimal after-ripening. Germination was under controlled environment at temperatures of 10, 15, 20 and 30 °C in darkness. A weighted germination index (WGI) was calculated. The analysis of WGI, for each temperature, showed highly significant (p ≤ 0.01) genotype effects on germination. Most genotypes decreased in WGI (increased dormancy) as temperature was increased from 10 to 30 °C. The greatest differences in seed germination tended to be at 15 °C and 20 °C. The level of seed dormancy depended on the genotype and germination temperature. This revised version was published online in August 2006 with corrections to the Cover Date.     

小麦种质多酚氧化酶的遗传变异及抗穂发芽品种的筛选

为了研究中国小麦品种资源多酚氧化酶(PPO)活性的变异,并对部分主推小麦品种的抗穗发芽能力进行评价和筛选,选用195份小麦种质资源和24份主推小麦品种,采用籽粒法测定了其多酚氧化酶（PPO）的活性。结果表明,供试小麦资源PPO活性表现出较大的变异,变异范围为4.95~13.99 AU/(min?g),变异系数为21.4%,表明小麦PPO活性遗传改良具有很大的潜力;我国主推小麦品种之间的PPO活性差异较大,筛选出‘中麦9’,‘京9428’,‘冀麦38’,‘陕229’,‘宁麦9’,‘川麦47’等PPO活性较低的品种,可以鉴定为抗穗发芽能力较强的品种。     

甘蓝型油菜成熟角果内源激素对带壳种子发芽的影响

以甘蓝型油菜品系832、780、1-1、1-13、8-6为材料,进行了田间带壳种子发芽调查、室内模拟降雨带壳种子发芽试验、种子发芽试验、不同浓度ABA处理的种子发芽试验,并利用HPLC测定了鲜果壳和种子的吲哚乙酸(IAA)、细胞分裂素(CK)和脱落酸(ABA)的含量。结果表明,各供试甘蓝型油菜品系带壳种子发芽率存在极显著差异,而不带壳种子的发芽率在第5 天以后没有显著差异。油菜成熟角果的果壳ABA含量与带壳种子发芽存在显著负相关（r＝－0.8941）,带壳种子发芽主要受果壳所含ABA的抑制;外源ABA能明显抑制种子发芽。     

小麦穗发芽抗性相关Vp1基因启动子的分离及功能验证

成熟期穗发芽严重影响小麦产量和品质。Vp1是调节胚发育, 促进胚成熟和休眠的重要转录因子, 对小麦种子休眠和穗发芽抗性具有重要作用。本研究分离了普通小麦B基因组Vp1基因的启动子, 生物信息学预测结果表明, 其含有9个脱落酸响应元件ABRE、2个DREB和6个MYB干旱响应元件、3个赤霉素响应元件GARE、1个水杨酸响应元件TCA-E、2个茉莉酸甲酯响应元件TGACG-motif、4个SKn-1和1个RYREPE胚乳特异表达元件。采用5′端缺失的方法, 构建了系列含Vp1启动子不同区段融合GUS报告基因的瞬时表达载体和植物表达载体。通过基因枪转化小麦愈伤组织, 瞬时表达结果显示, Vp1启动子在无诱导的情况下不能启动GUS基因表达, 在低温、ABA、GA、PEG和NaCl诱导后可以启动GUS基因表达, 表现诱导表达特性, 且其诱导表达强度随启动子缺失片段长度变短而减弱。利用Gateway方法成功构建了6个启动子各缺失片段类型的植物表达载体, 并通过农杆菌介导转化四倍体小麦Stewart, 获得转基因植株。该启动子可有效启动GUS基因在转基因植株的花药、糊粉层、穗轴及根中表达, 其他组织中没有表达。当启动子片段大于660 bp时, 外源ABA可诱导启动子启动GUS基因在转基因植株茎节中的表达。     

Breeding of white-grained wheats for Japan

The possibility of breeding white-grained wheats tolerant to pre-harvest sprouting under Japanese humid weather conditions is discussed. New genetic dormancy sources, such as, AUS1408, 8019R1 and RyuuMai7, were evaluated for seed dormancy in different weather conditions. Some white-grained dormant wheats showed a strong dormancy similar to that of red-grained dormant wheats in a greenhouse trial, in the field their dormancy expression was much less than the red wheats. Three populations involving crosses with these new sources for winter wheat breeding were examined under repeated selection for seed dormancy. Some dormant white-grained lines, as judged under glasshouse conditions, were developed. Again the level of dormancy in these lines was not sufficient compared to red dormant varieties in field trial. In order to develop truly superior dormant white-grained materials, one population involving crosses with two dormant varieties, AUS1408 and 8019R1, was examined under repeated selection. From these materials we succeeded in breeding lines that had not only a good dormancy but also showed adaptation to Japanese weather conditions, including earliness, scab resistance and good seed appearance. However again the dormancy of these lines in field trial was not sufficient compared to that of red wheats and there was not a clear difference for seed dormancy between breeding lines and their parents. We conclude therefore that more work involving the use of new genetic sources or new breeding techniques, will be necessary for breeding advanced lines that maintain a sufficient tolerance to PHS in humid Japanese weather condition. This revised version was published online in August 2006 with corrections to the Cover Date.     

Genetic analysis of pre-harvest sprouting in a durum wheat cross

Pre-harvest sprouting of durum wheat (Triticum turgidum L. var durum) reduces commercial grade, although the actual effects on processing quality are controversial. Little is known about the genetics of the dormancy component of pre-harvest sprouting resistance in durum. We studied the segregation of dormancy in 98 recombinant inbred lines from a cross of a relatively non-dormant line, CI13102, with a moderately dormant line, Kyle. The lines and parents were grown in field tests over three years, 1996, 1997 and 1998. Spikes were collected at approximately 20% moisture and stored at −23 ∘C. Hand-threshed grain of the lines was germinated, and number of seeds germinated was counted each day. A germination resistance index was calculated to characterize dormancy. Dormancy appeared to be complexly inherited in this cross. Lines were observed that were significantly (P < 0.05) more dormant than the parents. The lines transgressive for dormancy expressed in different combinations of the three environments, indicating an environmental interaction. DNA of lines and parents was tested with simple sequence repeat primers and AFLPs that were used in quantitative trait loci (QTL) analysis of dormancy. Significant QTLs for dormancy were found, with the most notable being on chromosome 1A, where other QTLs for pre-harvest sprouting resistance have been reported in common wheat.     

Mapping and validation of PCR-based markers associated with a major QTL for seed dormancy in wheat

Seed dormancy is one of the important factors controlling pre-harvest sprouting (PHS) resistance in wheat. We identified a major quantitative trait locus (QTL) for seed dormancy on the long arm of wheat chromosome 4A (4AL) via simple sequence repeat (SSR)-based genetic mapping using doubled haploid lines from a cross between Japanese PHS resistant variety ‘Kitamoe’ and the Alpine non-resistant variety “Münstertaler” (K/M). The QTL explained 43.3% of total phenotypic variation for seed dormancy under greenhouse conditions. SSR markers flanking the QTL were assigned to the chromosome long arm fraction length 0.59–0.66 on the basis of chromosome deletion analysis, suggesting that the gene(s) controlling seed dormancy are probably located within this region. Under greenhouse conditions, the QTL explained 28.5 and 39.0% of total phenotypic variation for seed dormancy in Haruyutaka/Leader (HT/L) and OS21-5/Haruyokoi (O/HK) populations, respectively. However, in field conditions, the effect was relatively low or not significant in both the K/M and HT/L populations. These markers were considered to be widely useful in common with various genetic backgrounds for improvement of seed dormancy through the use of marker-assisted selection. Further detailed research using near isogenic lines will be needed to define how this major QTL interacts with environmental conditions in our area.     

滇重楼种子休眠机理研究

探讨滇重楼种子休眠机理,为打破种子休眠促进滇重楼实生育苗提供理论基础。采用重量法、胚率测定、ELISA等方法对种子吸水特性、萌发抑制物及种胚休眠与发育的内源机理进行了研究。结果表明,滇重楼种子休眠属于形态学-生理学休眠类型。滇重楼种胚发育不完全、种子中存在萌发抑制物质是种子休眠的主要原因。种子不存在吸水障碍,与休眠无关。内源激素ABA和GA3在滇重楼种子休眠与萌发过程中起着重要作用,ABA是引起休眠的关键因素,ABA含量的降低是滇重楼种胚发育的始动因子,GA3有助于种子的生理后熟。萌发抑制物的存在可能抑制了种胚发育,而与生理学后熟过程无关。在自然条件下,滇重楼生境中较长的低温时期可能是滇重楼种子具有较长时间休眠的外界原因。     

Einfluß eines variierten N-Angebots auf α-Amylase-Aktivität, primäre Dormanz und Auswuchsresistenz von Weichweizen (Triticum aestivum L.), Roggen (Secale cereale L.) und Triticale (X Triticosecale Wittmack)

Influence of Varying N-Fertilization Rates on α-Amylase Activity, Primary Dormancy and Resistance to Pre-Harvest Sprouting in Wheat ( Triticum aestivum L.), Rye ( Secale cereale L.) and Triticale (X Triticosecale Wittmack)
Pre-harvest sprouting, induced by unfavourable ecological conditions, can affect the grain growers success considerably. Positive correlations are reported between resistance to pre-harvest sprouting and primary dormancy. Genotypes with a short dormancy period have a high pre-harvest sprouting risk. In the case of a premature germination of caryopses in the head of grain a hydrolysis of intact starch granules caused by the endoenzyme α-amylase takes place in the endosperm.
Negative correlations between falling number and protein content are reported, however, it is unknown, if a varying N-application influences pre-harvest sprouting rates, dormancy periods and amylase activity. For this reason, both greenhouse and field trials were conducted with different N-fertilization rates and (additional in the greenhouse) a rain simulation treatment.
High amounts of α-amylase a few days post anthesis are opposed to small enzyme activities in mature kernels. Stratificating temperatures and germination inducing precipitations at the same time are inducing pre-harvest sprouting and a high α-amylase activity especially in rye and triticale. It seems as if N-deficiency reduces the possibility of pre-harvest sprouting, on the other hand high N-rates increase the enzymes' activity and promote germination processes in the kernel. Effects of N-fertilization on dormancy are not known.
In the discussion of reasons for an increase of α-amylase activity in sprouted grain caryopses, changes in the relation of the phytohormones gibberellic acid (promoter of enzyme activities) and abscisine acid are mainly presumed.     

The effects of cis-trans ABA on embryo germination and seed dormancy in wheat

Pre-harvest sprouting of wheat grain can cause economic losses especially in cultivars with low levels of seed dormancy. The aim of this study was to determine genotype differences in embryo sensitivity to germination in response to exogenous (+/–) cis-trans ABA treatments at different concentrations. Six white and four red seed-colored bread wheat genotypes that differed in dormancy were grown in a field near Swift Current, Saskatchewan in 2000 as a randomized complete block design with four replicates. The seed samples from this experiment were germinated in a controlled environment at 20 °C without light. The exogenous ABA treatments were 0 μM – whole seed (control), 0 μM-embryos, 25 μM – embryos and 50 μM – embryos. The ABA experiment was a factorial design with four randomized complete blocks with four ABA treatments in all combinations with the ten genotypes. A weighted (by day) germination index (WGI) was calculated for each genotype in each ABA treatment. Genotypes differed in response to ABA. The genotypes, ABA concentration and genotype by ABA concentration interaction effects were significant (p ≤ 0.05). Excised embryos showed significantly decreased dormancy in most of the experimental genotypes. The addition of exogenous ABA enhanced embryo dormancy of most genotypes. This revised version was published online in August 2006 with corrections to the Cover Date.