A simple and efficient cryopreservation of in vitro-grown shoot tips of `Troyer' citrange [Poncirus trifoliata (L.) Raf. × Citrus sinensis (L.) Osbeck.] by encapsulation-vitrification

Wheat of two strong high-protein and two weak low-protein cultivars from New Zealand and Australia were milled to commercial specifications. All millstreams were tested for α-amylase, β-amylase, falling number, protein, starch, damaged starch, amylose, amylopectin, pentosan and ash. The distribution of β-amylase in millstream flours was more variable among cultivars than α-amylase. Generally, both enzymes had lowest activity in sizing and early reduction flours. α-Amylase was very high in the bran, pollard and germ fractions, in which ash content was very high, whereas β-amylase was low in these fractions. These observations, together with the moderate correlation of α-amylase and poor correlation of β-amylase to ash content, suggest that most α-amylasein flour derives from contamination with bran, pollard and germ, whereas most β-amylase derives from the endosperm. Falling numbers varied between the cultivars, but variation amongst millstreams for each cultivar was low, except for cv. Frame, which had particularly high falling number values (834 and 1197) in second and third break flours. These two flours had some of the highest α-amylase levels and lowest starch levels. However, they also had very high protein content (22 and 26%) and very low starch damage (3.2 and 4.5%), which may contribute to the high falling numbers. When endogenous α-amylase in the flour with the highest falling number was supplemented with high levels of barleyα-amylase, the flour withstood the detrimental effects of α-amylasein baking (sticky crumb, poor crumb texture and loaf volume) better than flours of lower falling number, but did not withstand the effects ofα-amylase on falling number. This revised version was published online in August 2006 with corrections to the Cover Date.     

Genetic variability of preharvest sprouting – the South African situation

Preharvest sprouting resistance is a major breeding criterion in many regions. Screening for preharvest sprouting tolerance is difficult owing to the low heritability of the trait and its tendency to be expressed as a quantitatively inherited character. The screening procedure used currently at the ARC-Small Grain Institute is designed to assess variation of sprouting in intact heads. Five winter wheat cultivars were crossed in a complete diallel fashion. The objectives of the study were to determine the genetic variability of sprouting resistance in the progeny of five winter wheat cultivars, including Elands, a cultivar with excellent sprouting resistance. The preharvest sprouting response and α-amylase activity of these cultivars and their ensuing progeny were compared. Elands was identified as contributing positively towards preharvest sprouting tolerance in various combinations. Elands also had the best overall combining ability for the improvement of sprouting tolerance.     

不同Wx蛋白缺失类型小麦穗发芽特性的初步研究

以11份Wx蛋白全部缺失类型（糯性），11份Wx蛋白部分缺失类型（非糯性）和大面积推广品种扬麦158、扬麦12（非糯性）为试验材料，以白皮品种豫麦47为穗发芽对照，研究了它们的穗发芽特性、α-淀粉酶活性和籽粒可溶性糖含量。结果表明，糯小麦的穗发芽率和籽粒可溶性糖含量均极显著高于非糯小麦，而α-淀粉酶活性与非糯小麦差异不显著。α-淀粉酶活性高是非糯小麦穗发芽的原因之一，两者呈指数关系，y=0.4145e^0.4864x(R²=0.3513*)；对于糯小麦，籽粒可溶性糖含量高可能是其穗发芽严重的诱导因素，两者亦呈指数关系，y=0.0001e^2.5599x(R²=0.5338**)。     

Regulation of high pI α-amylase synthesis in wheat aleurone by a gene(s) located on chromosome 6B

Synthesis of high and low pI α-amylase in germinating wheat grains and GA₃-treated de-embryonated grains of Chinese Spring (CS) and its ditelosomic derivative (CS dit 6BS) were compared and related to high pI α-amylase production in grains affected by late maturity α-amylase (LMA). In de-embryonated grains of CS dit 6BS (lacking the long arm of chromosome 6B) treated with GA₃, synthesis of high pI α-amylase isozymes controlled by Amy-1 genes on chromosomes 6A and 6D was dramatically reduced compared to Chinese Spring. The results suggest the presence of a gene(s) on the long arm of chromosome 6B, which is (are) required for GA-induced α-amylase synthesis in the aleurone. Similarly, in wheat grains affected by LMA, high pI α- Amy genes on the group 6 chromosomes are activated apparently by a single gene, tentatively located on chromosome 6B. Both genes may be part of the GA-induced amylase synthesis pathway in the aleurone. By contrast, synthesis of the high pI α-amylase isozymes in the early stages of germination of CS dit 6BS grains was very similar to CS. This contrast between GA₃-treated de-embryonated grains and germinating grains could be explained by control of α-amylase synthesis in scutellum in the early stages of germination being different to that in GA₃-treated aleurone. This revised version was published online in August 2006 with corrections to the Cover Date.     

Characterization of preharvest sprouting resistance in Clark's Cream white winter wheat

Summary Resistance to preharvest sprouting has been observed in several white wheat (Triticum aestivum L.) genotypes, but the mode of resistance has not been determined. Studies were conducted to characterize the strong preharvest sprouting resistance in Clark's Cream genotype to facilitate breeding for the trait. Clark's Cream and five other hard red and white wheat genotypes were grown in the glasshouse and field and effects of simulated rain, embryo maturity, inhibitors in floral parts and GA, endogenous inhibitors, and ABA and GA combinations on sprouting were measured. The six genotypes differed significantly in sprouting after simulated rain, -amylase activity, and response to GA and ABA. Embryo maturity and inhibition levels in floral parts and caryopses were nearly similar for all genotypes. Clark's Cream appeared to differ most in high embryo sensitivity, which was fairly nonspecific for ABA, GA, and endogenous inhibitor, and in low -amylase production during sprouting. Breeding procedures that lead to the preharvest sprouting resistance of Clark's Cream are discussed.     

Inheritance of late maturity α-amylase in wheat

Summary Two wheat cultivars that consistently show high levels of grain -amylase at harvest ripeness, in the absence of preharvest sprouting, were crossed with a control, low -amylase cultivar, and F₁, F₂ and BC₁ populations were developed. Grain of these populations was analysed for -amylase activity at harvest ripeness. Distribution and segregation patterns were consistent with control at a single locus with high -amylase the recessive allele. This mode of inheritance would make it extremely difficult to differentiate homozygous low -amylase lines from heterozygotes (low -amylase phenotype but carriers of high -amylase) and has important implications for wheat breeders. High -amylase, termed late maturity -amylase, was not linked with the awned inhibitor gene, B2, located on the long arm of chromsome 6B.     

Gibberellic acid effects on germination and α-amylase activity of winter wheats

Summary Sensitivity to GA in non-Gai genome winter wheat (Triticum aestivum L.) cultivars was investigated to determine magnitude of variation of the trait, its association with other traits, and effects of geographical location of production. -Amylase enzyme activity was measured before and after treatment with gibberellic acid in 18 cultivars grown at one location and in five cultivars grown at six locations. Dye-labeled starch and agar-starch media procedures were used for the first and second experiments, respectively. -Amylase activity differed significantly among the 18 cultivars after germination in water, germination in GA, and in absolute and relative sensitivity to GA. Cultivars that reacted similarly to GA had some common ancestors in their pedigrees, and -amylase activity and sensitivity to GA were significantly negatively associated with seed weight. -Amylase activity differed between tall and semi-dwarf cultivars in the second experiment, but not in the first experiment. The magnitude of variation in -amylase activity and its relationship to preharvest sprouting susceptibility of the cultivars suggested that the trait can be modified to improve seed dormancy. Significant interactions between cultivar responses to GA and geographical location of grain production suggested that selection should be carried out in several environments.Contribution no. 81-162-j, Department of Agronomy, Kansas State University, Manhattan KS 66506, USA.     

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.     

Screening methods and identification of QTLs associated with late maturity α-amylase in wheat

The synthesis of high levels of germination-type (high pI isozymes) α-amylase was induced in wheat genotypes prone to late maturityα-amylase (LMA) following the exposure of detached tillers to cool temperature during grain development. The detached tiller method was successfully applied to a range of genotypes and to a doubled haploid (DH) population derived from the cross Cranbrook (LMA genotype) × Halberd (low amylase). The number of grains in ripe, treated tillers that contained high pI α-amylase isozymes was measured using an ELISA antibody kit highly specific for high pI isozymes. Quantitative trait loci (QTL) controlling the expression of LMA in wheat were detected in DH population Cranbrook × Halberd. The DH population and parents were sown in 2 replicated sowings at the same location with sowing times differing by 2 weeks. QTL analyses were conducted separately for each sowing, but results from both sowings were consistent and indicated a highly significant (it p > 0.01) QTL on the long arm of chromosome7B, with Cranbrook contributing the higher value allele. A second QTL with less significant effect was found on the long arm of chromosome 3B, on the basis of data from the first sowing. This revised version was published online in August 2006 with corrections to the Cover Date.     

Breeding for salinity resistance in mulberry (<Emphasis Type="Italic">Morus</Emphasis> spp.)

Late maturity α-amylase (LMA) is a genetic defect that is fairly widely spread in bread wheat (Triticum aestivum L.) germplasm, and recently detected in durum cultivars, which can result in unacceptably high α-amylase activity (low falling number) in ripe grain. LMA has also been observed at unexpectedly high frequency and severity in synthetic hexaploid wheats derived from the interspecific hybridisation of Triticum durum (AABB) and Aegilops tauschii (DD). Since synthetic hexaploids represent an important new source of resistances/tolerances to a range of biotic and abiotic stresses for wheat breeders, there is a pressing need to understand the mechanisms involved in LMA in synthetics and develop strategies for avoiding its adverse effects on grain quality. The objectives of this study were to firstly, compare the LMA phenotype of synthetics that varied for plant height, secondly, to characterise the LMA phenotype in groups of synthetics derived from the same durum parents and finally to determine whether LMA in primary synthetics is associated with the QTL previously reported in conventional bread wheat. More than 250 synthetic hexaploids, a range of durum cultivars and a doubled haploid population derived from Worrakatta (non-LMA) × AUS29663 (high LMA synthetic) were phenotyped and genotyped with markers reported to be linked to LMA in conventional bread wheat and markers diagnostic for the semi-dwarfing gene, Rht1. More than 85% of synthetics were prone to LMA, approximately 60% ranked as very high. Genetic control of LMA in synthetic hexaploids appeared to involve QTL located on 7B, and to a lesser extent 3B, similar to bread wheats. However, the LMA phenotype of many synthetic hexaploids appeared to be more extreme than could be explained by comparisons with bread wheat even taking into account the apparent absence of Rht1 in most genotypes. Other mechanisms, possibly triggered by the interaction between the AABB and DD genomes cannot be excluded. The presence of wild type rht1 in most synthetic hexaploids and their extreme height is difficult to reconcile with the semi-dwarf, Rht1, stature of many of the durums used in the interspecific hybridisation process. Mechanisms that could explain this observation remain unclear.     

中国小麦品种穗发芽抗性差异的研究

利用收获时种子发芽率和面粉降落值法，于2000—2002年2个小麦种植年度，研究了黄淮、北部、长江中下游、西南冬麦区和东北春麦区1950年以来的781个主要推广品种和新品系的穗发芽抗性。结果表明，小麦穗发芽抗性测定值在年度间极显著或显著正相关，不同年代小麦品种的穗发芽抗性差异较大。1990年以来育成品种的穗发芽抗性与20世纪80年代相近，但明显弱于50、60以及70年代。黄淮、西南和春小麦3个麦区种子发芽率低于2%的高抗品种数分别占各自麦区供试总数的1.7%、4.5%和5.7%，而长江中下游和北部2个冬麦区的种子发芽率都在10%以上；东北春麦区品种的抗性较强，种子发芽率平均为11.2%。利用等电聚焦电泳从发芽率和降落值均偏低的品种中鉴定出异源2号、蜀万24、蜀万761、陕160、孟县4号、京411、京9428、鉴26、燕大1817、农大45、衡水6404、晋麦5号、8号、鄂麦14和克辉等15个携带迟熟α-淀粉酶基因的品种，分布在5个不同麦区。对这些品种及其亲本进行SSR分子标记分析，发现有些与其亲缘关系密切的品种，却不携带迟熟α-淀粉酶基因。上述结果说明，该基因在我国五大小麦产区均有分布，但具该基因的品种数量少，占供试品种数的1.9%，通过育种程序容易选择出不携带该基因的小麦品种。     

Pre-maturity α-amylase and incipient sprouting in UK winter wheat,with special reference to the variety Rialto

In 1998 and 1999 the UK winter wheat variety Rialto produced unexpected low Hagberg falling numbers that could not be directly linked to sprouting. It was proposed that these reductions in quality could be due to pre-maturity α-amylase activity (PMAA). The problem was not identified during the selection and commercial development stages. Our study tested the hypothesis that the variety Rialto is PMAA-susceptible. Analysis was done on 13 year-location combinations of field grown Rialto. Together, visual and chemical assessments of sprouting and iso-electric focusing of α-amylase isozymes identified several samples with significant α-amylase activity in the apparent absence of sprouting. In addition, tests with α-amylase sensitive Phadebas gel revealed distinctive PMAA discoloration patterns in 10–44% of the grain from the 13 samples, leading to the conclusion that Rialto is PMAA-susceptible. Diurnal temperature range accumulated for an 11 day period during a warm spell in early simulated grain development displayed a significant but negative correlation with the number of grains showing clear PMAA discoloration patterns on Phadebas gel. The number of clear PMAA grains correlated positively with rainfall accumulated over 11 days. These results suggest that PMAA can increase under conditions similar to those conducive to pre-harvest sprouting. It is however also possible that in some instances both PMAA and incipient sprouting could have produced similar patterns of α-amylase activity. In addition to tests with Rialto, Phadebas gel tests were therefore also done with the known high Hagberg varieties Option and Malacca, sprouted in a controlled environment. Results from the additional gel tests suggest that visual and chemical assessments of sprouting in the grain combined with Phadebas gel analysis could identify PMAA more reliably in grain sub-samples than Phadebas gel analysis alone.     

Heritabilities and genetic variation for preharvest sprouting in progenies of Clark's Cream white winter wheat

Summary Preharvest sprouting resistance is a major breeding criterion in many regions where white wheat (Triticum aestivum L.) is produced. Availability of genetic sources of sprouting resistance should significantly advance this goal. Objectives of the present investigation were to determine heritabilities and genetic variation of sprouting resistance in progeny of six susceptible genotypes and Clark's Cream, a cultivar with excellent sprouting resistance. Sixty-five randomly selected F₂:F₃ lines from the six parental combinations were evaluated in blocks-within-replication designs at two locations during 1984–85. Falling number, -amylase activity, and sprouting percentage in simulated rain were usually more favorable in F₂:F₄ grain than in grain of the susceptible parents and frequently equalled levels in grain of the resistant parent. Broad sense heritability estimates were moderate to high for falling number and -amylase activity, low to moderate for visual sprouting, and inconsistent for embryo germination in ABA. Phenotypic correlations with sprouting were significant most often for falling number and least often for the agronomic traits, days to heading and kernel weight. We concluded that increased preharvest sprouting resistance is an attainable objective when genetic sources of strong resistance and appropriate selection criteria, such as sprouting after simulated rain, are used.     

Exploiting the diversity of <Emphasis Type="Italic">Viviparous-1</Emphasis> gene associated with pre-harvest sprouting tolerance in European wheat varieties

Pre-harvest sprouting (PHS) reduces the quality of wheat (Triticum aestivum L.) and the economic value of the grain. The objective of this study was to evaluate the diversity of the Viviparous-1B (Vp-1B) gene associated with PHS tolerance in a collection of 490 widely grown winter wheat varieties from central and northern Europe. Four alleles of Vp-1B were found in the wheat varieties tested, three of which (Vp-1Ba, Vp-1Bb and Vp-1Bc) had previously been identified in Chinese wheat varieties. The fourth was a new allele which had a 25-bp of deletion in the third intron region compared with the nucleotide sequence of Vp-1Ba, and was designated as Vp-1Bd. The frequencies of different alleles in this set of European wheat germplasm were: Vp-1Ba (54%) > Vp-1Bc (21%) > Vp-1Bd (20%) > Vp-1Ba + c (4%) > Vp-1Bb (1%), with Vp-1Bb being present only in two French varieties, ‘Altria’ and ‘Recital’. In addition, the frequencies of the alleles differed in varieties from different European countries. For example, Vp-1Ba had the highest frequency (76%) in varieties included in the UK National List (NL), but was least frequent in the Recommended List (RL) of Sweden (19%). Similarly, Vp-1Bc was present with the highest frequency (58%) in wheat varieties from Sweden, and the lowest in UK NL varieties (8%) while Vp-1Bd had the highest frequency of 32% in German varieties, and the lowest in Sweden varieties with only 8%. The Vp-1Ba allele was present in over half of the UK wheat varieties tested but the frequency was lower in RL varieties than in NL ones. Furthermore, heterogeneities were found between Vp-1Ba and Vp-1Bc in the varieties from Sweden, Netherlands, Germany and UK.     

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.     

Utilization of tom thumb source of pre-harvest sprouting tolerance in a wheat breeding programme

Summary Segegating populations of the cross Tordo (a spring white-grained Tom Thumb derivative) and Tr 454-16 (a spring white-grained Sonora-Gabo derivative) were studied for plant height, apparent -amylase synthesis (AAS) and response to gibberellic acid. Means, variances and heritabilities for these characters were computed and their association was studied by computing correlation coefficients. Plant height showed high heritability while AAS and response to GA₃ showed moderate heritability. Plant height was positively correlated with AAS and response to GA₃. The results pointed out the importance of using Tom Thumb as a source of pre-harvest sprouting tolerance in a wheat breeding programme, particularly in areas where wheat breeding is restricted to evolving white-grained cultivars.     

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。     

More precise map position and origin of a durable non-specific adult plant disease resistance against stripe rust (Puccinia striiformis) in wheat

Recently a major gene determining non-specific adult plant disease resistance against stripe rust (Puccinia striiformis) designated Yrns-B1 was mapped in wheat Triticum aestivum L. by using a cross between ‘Lgst. 79-74’ (resistant) and ‘Winzi’ (susceptible). Linkage to five Gatersleben wheat microsatellite (GWM) markers was discovered, previously mapped on chromosome arm 3BS. In the present study this map was improved by the incorporation of four additional GWM markers. QTL-analysis revealed high LOD values for the resistance at all nine loci, whereas the largest LOD (20.76) was found for the newly mapped marker Xgwm1329. Microsatellite analysis and resistance tests of a collection of old German/UK wheat varieties, including probable ancestors of ‘Lgst.79-74’ were carried out. A high coincidence of non-specific adult plant disease resistance against stripe rust and the presence of ‘Lgst. 79-74’ allele (117 bp) of the marker Xgwm533 was observed among the varieties tested. Linkage during the inheritance of both the resistance and the 117 bp allele of Xgwm533 was demonstrated. The probable origin of Yrns-B1 is discussed. Carriers of this resistance gene were grown on large areas since more than 100 years. To estimate the capability of Xgwm533 as a diagnostic marker for non-specific adult plant disease resistance against stripe rust, microsatellite analysis and resistance tests of a collection of Russian spring wheat varieties were performed. The 117 bp allele of Xgwm533 was found in about 35% of the Russian cultivars analysed, however, none of them possessed the expected disease resistance. Thus, the utilisation of Xgwm533 as diagnostic marker seems to be restricted to certain genepools.     

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.     

Expression of late maturity α-amylase in wheat containing gibberellic acid insensitivity genes

Summary Two wheat cultivars, Spica and Lerma 52, which consistently produce high levels of -amylase during the later stages of grain development (late maturity -amylase), were crossed with a set of four near-isogenic lines carrying the tall (rht) allele or one of the dwarfing genes Rht1, Rht2 or Rht3 (GA-insensitive alleles). The F₁ and F₂ populations were developed and analysed for grain -amylase and plant height. The Rht3 gene exhibited the strongest influence on plant height and strongly inhibited new -amylase synthesis during the later part of grain ripening. By comparison, Rht1 and Rht2 had a less pronounced effect but still significantly reduced the expression of late maturity -amylase. These observations suggest that gibberellic acid is involved either directly or indirectly in this phenomenon. The implications of the effect of dwarfing genes on expression of late maturity -amylase are discussed in relation to cultivar improvement and to the identification and control of high -amylase germplasm.