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.     

Response of Wheat Grain to ABA and Imbibition at Low Temperature

During after-ripening of wheat grains, embryo responsiveness to abscisic acid (ABA) decreases in parallel with the loss of grain dormancy. Dormant grains of Triticum aestivum L. line ‘Kitakei-1354’ that had lost almost half their dormancy due to chilling still respond to ABA like fully dormant grains. Imbibition a: low temperature appears to break the dormancy without change of embryo responsiveness to ABA. Gibberellin (GA) and ABA-regulated α-amylase synthesis of the embryos. This is amplified when the grains are imbibed at low temperature. Imbibition at low temperature appears to condition cells and tissues of the embryo for germination and α-amylase synthesis.     

Wheat ABA-insensitive mutants result in reduced grain dormancy

This paper describes the isolation of wheat mutants in the hard red spring Scarlet resulting in reduced sensitivity to the plant hormone abscisic acid (ABA) during seed germination. ABA induces seed dormancy during embryo maturation and inhibits the germination of mature seeds. Wheat sensitivity to ABA gradually decreases with dry after-ripening. Scarlet grain normally fails to germinate when fully dormant, shows ABA sensitive germination when partially after-ripened, and becomes ABA insensitive when after-ripened for 8?C12?months. Scarlet ABA-insensitive (ScABI) mutants were isolated based on the ability to germinate on 5???M ABA after only 3?weeks of after-ripening, a condition under which Scarlet would fail to germinate. Six independent seed-specific mutants were recovered. ScABI1, ScABI2, ScABI3 and ScABI4 are able to germinate more efficiently than Scarlet at up to 25???M ABA. The two strongest ABA insensitive lines, ScABI3 and ScABI4, both proved to be partly dominant suggesting that they result from gain-of-function mutations. The ScABI1, ScABI2, ScABI3, ScABI4, and ScABI5 mutants after-ripen more rapidly than Scarlet. Thus, ABA insensitivity is associated with decreased grain dormancy in Scarlet wheat. This suggests that ABA sensitivity is an important factor controlling grain dormancy in wheat, a trait that impacts seedling emergence and pre-harvest sprouting resistance.     

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。     

Relationship between sprouting in wheat and embryo response to endogenous inhibition

Summary Seed dormancy in wheat (Triticum aestivum L. em Thell) is important for minimizing pre-harvest sprouting. To facilitate breeding cultivars that tolerate pre-harvest sprouting conditions, we assessed mode and magnitude of variation of seed dormancy among genotypes and investigated involvement of endogenous water-soluble inhibitor(s) in seed germination. Embryo bio-assays established that water-soluble inhibitor was ubiquitous among the wheat cultivars studied and did not diminish in quantity during after-ripening. Germination response of embryos was decreased by endogenous inhibitor, but the effect markedly declined as embryos aged at room temperature. Variation in dormancy among cultivars was primarily caused by differential response of their embryos to endogenous inhibitor. Gibberellic acid counteracted the initial inhibitory effect of endogenous inhibitor on germination but not the subsequent inhibitory effect on seedling growth. We concluded that pre-harvest sprouting resistance involves multiple factors, particularly embryo receptivity to endogenous inhibitor, and that variation in inhibitor quantity is not solely responsible for genotypic differences in susceptibility to pre-harvest sprouting. The possibility of additional approaches to breeding for pre-harvest sprouting resistance is indicated.Contribution no. 81-389-j, Department of Agronomy, Kansas State University, Manhattan, KS 66506, U.S.A.     

Mapping a diploid wheat abscisic acid 8′-hydroxylase homologue in the seed dormancy QTL region on chromosome 5A<Superscript>m</Superscript>

In Arabidopsis, two genes of abscisic acid (ABA) 8′-hydroxylase (cytochrome P450 (CYP) 707A1 and A2) have been found to play important roles in seed dormancy through the regulation of ABA content in seeds. In order to examine the role of wheat ABA 8′-hydroxylase gene in seed dormancy, a diploid wheat ABA 8′-hydroxylase gene was cloned that showed high similarity to a barley ABA8′-hydroxylase gene (HvABA8′OH-2), and the cloned gene was designated as TmABA8′OH-2. Using recombinant inbred lines derived from a cross between diploid wheat Triticum boeoticum L. (Tb) and Triticum monococcum L. (Tm), TmABA8′OH-2 has been mapped to near the centromeric region of the long arm of chromosome 5A^m, where the major seed dormancy QTL has been previously found. Comparison of the deduced amino acid sequences of TmABA8′OH-2 between Tb and Tm revealed five amino acid residue substitutions. These amino acid residues have distinctly different characteristics, and one of the substitutions occurs in the highly conserved amino acid residues in CYP707A family, indicating that these substitutions may have effects on the enzyme activities. Moreover, hexaploid wheat TmABA8′OH-2 homologue revealed that the level of its expression during seed development peaks at mid-maturation stage. This resembles the expression pattern of the Arabidopsis CYP707A1, which was shown to control seed dormancy. These results imply a possibility that TmABA8′OH-2 might be involved in seed dormancy, and associated with the QTL on chromosome 5A^m.     

花生种子休眠解除过程中相关基因的表达分析

种子休眠性是花生重要的农艺性状,外源乙烯利能诱导花生种子休眠的解除,为了阐明乙烯利作用下花生种子休眠解除的分子机制,设置吸胀的休眠种子为对照,100 mg L–1乙烯利处理吸胀休眠种子后不同时间的样品(AE1、AE2、AE3)进行转录组分析,比较了花生种子休眠解除过程中ABA、GA、ETH、auxin相关基因的表达。结果表明,15个与GA、40个与ABA、60个与ETH、56个与auxin相关的unigenes在花生种子休眠解除过程中表现显著差异表达。荧光定量PCR结果显示,ABA合成关键基因Ah NCED2和代谢关键基因Ah CYP707A1在种子休眠解除过程中均受外源乙烯利诱导,表达差异显著;在休眠和无休眠种子吸胀萌发过程中,Ah NCED2和Ah CYP707A1的表达趋势不同,Ah NCED2对于种子休眠的维持发挥积极作用,而Ah CYP707A1对于种子休眠解除发挥积极作用。     

Response of maize inbred lines to a defoliation treatment inducing tolerance to cold at germination

Defoliation during maize (Zea mays L.) kernel development has been observed to induce tolerance to cold of germinating seeds in responsive genotypes. The objectives of this study were to evaluate the response to defoliation of immature embryo and mature seed germinability at cold and to verify if the response was influenced by the developmental stage at which the treatment was applied. In three environments, six inbred lines (B73, IABO78, Lo1016, Lo964, Mo17, Os420) were defoliated (D) approximately 20 days after pollination (DAP) or not defoliated (ND). Immature embryos were excised three days after defoliation and germinated in vitro at 9 or 25 ^∘C. At maturation, kernel germination was tested at the same temperatures. Defoliation improved cold tolerance and mean time to germination (MTG) at 9 ^∘C of both embryos and kernels of Lo1016. To study the effect of kernel developmental stage on response to defoliation, plants of B73, Lo1016 and Lo964 were defoliated at 15, 18, 21, 24, 27, 30, 33, 36, and 39 DAP, or not defoliated. At the same DAP, immature grains were analyzed for dry weight, water and abscisic acid (ABA) content. In Lo1016, low amounts of kernel ABA were detected at all stages, while in Lo964 and B73 ABA increased during development. Lo1016 mature kernels showed an improvement of cold tolerance due to defoliation at all times, while the other genotypes did not. In conclusion inbred lines showed variability for mature seed and immature embryo tolerance to cold at germination and for the ability to acquire tolerance after defoliation.     

Exogenous Abscisic Acid Application During Grain Filling in Winter Wheat Improves Cold Tolerance of Offspring's Seedlings

Low temperature seriously depresses seed germination and seedling growth in winter wheat (Triticum aestivum L.). In this study, wheat plants were sprayed with abscisic acid (ABA) and fluridone (inhibitor of ABA biosynthesis) at 19 days after anthesis (DAA) and repeated at 26 DAA. The seeds of those plants were harvested, and seed germination and offspring's seedling growth under low temperature were evaluated. The results showed that exogenous ABA application decreased seed weight and slightly reduced seed set and seed number per spike. Under low temperature, seeds from ABA‐treated plants showed reduced germination rate, germination index, growth of radicle and coleoptile, amylase activity and depressed starch degradation as compared with seeds from non‐ABA‐treated plants; however, activities of the antioxidant enzymes in both germinating seeds and seedling were enhanced from those exposed to exogenous ABA, resulting in much lowered malondialdehyde (MDA) and H₂O₂ concentrations and production rate. In addition, the maximum quantum efficiency of photosystem II was also enhanced in ABA‐treated offspring's seedlings. It is concluded that exogenous ABA treatment at later grain‐filling stage could be an effective approach to improve cold tolerance of the offspring during seed germinating and seedlings establishment in winter wheat.     

The effect of germination temperature on seed dormancy in Croatian-grown winter wheats

The expression of seed dormancy related to germination temperature was studied in 25 wheat genotypes grown in the field at two locations near Zagreb and ?upanja in Croatia during 2008/2009 growing season. Germination tests were conducted at 15, 20 and 25?°C at harvest maturity (Time 1) as well as after 10?days (Time 2) and 15?months (Time 3) of seed after-ripening at room temperature, respectively. Significant (P?<?0.05) differences among locations (L), temperatures (T) and genotypes (G) as well as significant L?×?T, G?×?L, G?×?T and G?×?L?×?T interactions were observed for weighted germination index (WGI) at both Time 1 and Time 2. At Time 3 significant differences among genotypes for germination percentage were found only at the early stages of germination. The 25 wheat genotypes responded with decreasing WGI mean values (increasing dormancy) as temperature changed from 15 to 25?°C. The rate of dormancy increase with higher germination temperature varied among genotypes. Some genotypes, having similar values of WGI at 15?°C, significantly differed from each other at 25?°C and vice versa. This indicates that the range of germination temperatures included in the present study is useful when testing genotypes for their temperature-dependent dormancy potential. The number of genotypes with WGI values significantly different from the mean, as a measure of the power of germination test to detect differences in dormancy level among genotypes, as well as heritability estimates for WGIs were the highest at Time 1 for 15?°C and at Time 2 for 20?°C.     

外源生长调节物质对阔叶杨桐种子萌发的影响

试验研究了厚皮香科植物阔叶杨桐种子在3种外源生长调节物质GA、IAA和6-BA浸种处理下的萌发特性。结果表明,阔叶杨桐种子为休眠性种子,不同外源生长调节物质对种子萌发影响不同,与对照相比,50 mg/L和100 mg/L的GA浸种提高了种子的萌发率,而150 mg/L的GA处理抑制种子的正常萌发;低浓度的IAA(<100 mg/L)浸种处理促进种子的萌发,高浓度IAA浸种降低种子的萌发率;种子萌发对不同浓度6-BA的响应趋势与IAA相似,综合分析认为,低浓度的外源生长调节物质浸种处理可以在一定程度上解除种子的休眠,提高种子的萌发率,其中50 mg/L的6-BA浸种处理12 h效果最佳。     

The effects of parthenogenesis on wheat embryo formation and haploid production with and without maize pollination

Doubled haploid (DH) lines are important in wheat (Triticum aestivum L.) breeding, and haploids produced via maize pollination precede DH line development. Although maize pollination has proven reliable and broadly applicable to wheat, its success is determined by the wheat and maize genotypes employed. A wheat genotype consisting of nuclear and cytoplasm components predisposing it to parthenogenesis was compared with three other genotypes, each possessing only one or neither component necessary for parthenogenesis. In a glasshouse experiment, each genotype was pollinated with maize and subsequently treated with a2,4-Dichlorophenoxyacetic Acid (2,4-D) solution to determine if parthenogenesis affected embryo formation frequency (EFF)and haploid formation efficiency (HFE). Wheat genotypes were also treated with the2,4-D solution alone to determine if embryos and haploid plants could be produced in vivo without maize pollination. ‘Salmon(K)’, a parthenogenetic genotype consisting of a Salmon 1BL.1RS nucleus in a Ae. kotschyii cytoplasm, had a mean EFF of 32%; whereas, the non-parthenogenetic genotypes had mean EFF calculations ranging from 7 to 21%. Mean HFE for Salmon(K) was not significantly different than the mean HFE for non-parthenogenetic Salmon; however, EFF and HFE calculations for Salmon(K) and Salmon, each with a 1BL.1RS translocation, were generally higher than calculations for genotypes without the translocation. Salmon(K) was the only genotype to produce a 3% or higher EFF and HFE after treatment with 2,4-D alone. Parthenogenesis significantly affected the frequency at which embryos were produced after pollination with maize and the frequency at which embryos and haploid plants were produced after treatment with 2,4-D alone. This revised version was published online in July 2006 with corrections to the Cover Date.     

Chromosomes responsible for sensitivity of embryo to abscisic acid and dormancy in wheat

The sensitivity of the embryo to abscisic acid (ABA) has been reported toplay an important role in seed dormancy. Using ditelocentric lines of wheatcv. Chinese Spring (CS, nondormant and ABA insensitive), F₂ seedsbetween monosomic lines of CS and a wheat line Kitakei-1354 (dormant,ABA sensitive) and deletion lines of CS chromosome 4A, germinability ofseeds and embryo-half seeds incubated in water and ABA were examined. The results indicated that the long arm of chromosome 4A carried majorgene(s) for the embryo sensitivity to ABA and dormancy. Chromosome2D might be also involved in the sensitivity to ABA.     

Changes in wheat seed storage protein fingerprint due to soil mineral content

Wheat seed storage protein fingerprint is used to determine the gluten protein pattern in studies aimed at improving flour quality. Wild wheat with high seed protein content is used extensively in wheat breeding programs. Although the wild wheat growth and protein content may be influenced by environmental conditions, the gluten-protein pattern is generally considered as indicative of a genotype, without the superimposition of environmental influences. The effects of soil type, habitat, and deficiencies of N, P, K and S on seed storage protein composition were examined in nine accessions of wild wheat (Triticum turgidum var. dicoccoides) and three varieties (two T. aestivum and one T. durum). Soil from ten natural habitats of the wild wheat that had not previously received any fertilizers or manures was sampled and used to grow wheat in a greenhouse. Seed storage protein composition was characterized by SDS-PAGE. Although deficiencies in soil nutrient caused variations in the seed storage proteins, the genotype was the main factor determining the seed storage protein composition. Seed storage protein composition of genotypes varied when grown under different mineral nutrient conditions. Only one genotype was stable showing almost identical protein patterns under all growing conditions studied without any qualitative change in fingerprint pattern. In the other genotypes, as well as the cultivars, the seed storage protein was affected at least to some extent by the soil. The ‘soil effect’ is summarized in terms of three main quantitative changes in the seeds: 1 – the relative amounts of the high-molecular-weight proteins; 2 – the relative amounts of proteins in the range of 45 and 65 kD; 3 – the percentage distribution of the HMW glutenin and other groups of seed storage proteins. The soild induced also qualitative differences in the composition of seed storage proteins, mostly in those of 45–65 kD. These differences were observed whenever a deficiency of S, N, P, K or Mg was identified. Therefore, in breeding programs that use seed storage protein fingerprints of wild wheat germplasms should be exercise caution when the germplasms selected from wild habitats. This revised version was published online in July 2006 with corrections to the Cover Date.     

Wild wheat adaptation in different soil ecosystems as expressed in the mineral concentration of the seeds

Wild emmer wheat, Triticum dicoccoides, grows naturally in several habitats in northern Israel. The assumption that a genotype is better adapted to the `native' soil from which it was collected than to other soils, was tested. Each of nine T. dicoccoides accessions from nine different habitats and three wheat cultivars was sown in soils taken from all habitats, and grown in a greenhouse over 3 years. To enhance the biological nutrition absorption forces from the soil, three common wheat cultivars were added to the wild genotypes. No interaction in grain yield between wild wheat genotype and soil type was found within experiments. Soil type was the main factor that affected development and yield. Seed nutrient ability (SNA) of each soil was defined as the mineral element content in the seeds, averaged over all genotypes. Multiple regression analysis revealed diversity between the SNA related to growth and yield of the genotypes. Total seed yield per plant of each accession was related to several SNAs, and mainly to S and K, R² = 0.5–0.85. The spikelet number per spike was determined by N and Na in five accessions and by Ca in the other four (R² = 0.39–0.93).Heading date was affected mainly by the genotype, and the soil effect exhibited Fe and P dependence. A genotype-habitat adaptation exhibited by yield components was related to yield quality rather than to yield quantity. When a mineral nutrient is deficient in a natural soil, natural selection leads to establishment of plants that store a higher concentration of that nutrient in the seed, for the benefit of the succeeding generation. This revised version was published online in July 2006 with corrections to the Cover Date.     

Drought and high temperature increases preharvest sprouting tolerance in a genotype without grain dormancy

Preharvest sprouting is common in cereals, which lack grain dormancy when maturing grain is exposed to rainfall or high moisture conditions. Environmental conditions such as drought and high temperature during grain filling have a large effect on the expression of sprouting tolerance. A dormant (DM 2001) and non-dormant (Cunderdin) hard white spring wheat were exposed to drought or irrigated conditions and either low or high temperature during grain filling. Dormancy and embryo sensitivity to ABA were analysed throughout grain filling. The conclusions from this investigation were as follows; firstly DM 2001 was more dormant than Cunderdin, with a four-fold lower germination index (GI) at maturity. Secondly during grain ripening drought increased dormancy and overrides any increase in dormancy with low temperature. Finally embryo sensitivity can be induced in a non-dormant genotype to the extent where the non-dormant genotype in a hot dry environment can have the same phenotype as a dormant genotype grown in a cool wet environment. In summary drought during grain filling increases dormancy suggesting breeders need to avoid drought when screening for sprouting tolerance in order to maximise the chances of identifying genetic differences in grain dormancy and avoid any maturity by drought interactions.     

New genetic sources of resistance in the genus <Emphasis Type="Italic">Phaseolus</Emphasis> to individual and combined aluminium toxicity and progressive soil drying stresses

Bean species and genotypes show wide phenotypic variability in relation to aluminium (Al) resistance and progressive soil drying. The objective of this study was to identify and characterize sources of resistance to Al toxicity and progressive soil drying among six genotypes of common bean (Phaseolus vulgaris), four of runner bean (P. coccineus), and one of tepary bean (P. acutifolius), using hydroponic and soil cylinder screening methods. One experiment on hydroponic screening of Al resistance was carried out using a basal nutrient solution with and without 20 μM Al. Two experiments were carried out using two oxisols in 80 cm long soil cylinders with high Al (HAl) and low Al (LAl) saturation treatments. The three experiments showed an average of 36.9–53.5% inhibition of root growth with HAl compared with LAl treatments. Differences in root development and distribution were observed among genotypes and species. Two accessions of P. coccineus (G35346-2Q, G35464-5Q) and one Andean common bean genotype (ICA Quimbaya) were outstanding in root and shoot growth in the HAl treatments. P. coccineus accession (G35346-3Q) was outstanding under combined stress of Al-toxic acid soil and progressive soil drying. Accessions of P. coccineus may represent unique sources of Al resistance for the improvement of common bean through interspecific crosses.     

Effect of Abscisic Acid on Pollen Germination and Tube Growth of Maize Genotypes1

As a preliminary step to using gametophytic selection to obtain genotypes of maize, Zea mays L., differing for abscisic acid (ABA) content and/or sensitivity, the influence of ABA on the in vitro germination and tube growth of maize pollen and the presence of genetic variability for these traits was investigated. In a first experiment, pollen samples of three genotypes were germinated on substrates containing increasing amounts of ABA. Low ABA concentration (0.5 νM) significantly increased pollen germination and tube growth, while 500 νM ABA significantly decreased both traits, thus indicating that ABA can influence the growth of maize pollen in vitro. In a second experiment, eleven inbred lines were screened for the in vitro reactivity of their microgametophytes to ABA. The lines differed significantly for the response (% of control) of tube length, but not of pollen germination to ABA.     

Antimicrotubule herbicides for in vitro chromosome doubling in Beta vulgaris L. ovule culture

In vitro chromosome doubling during ovule culture of sugar and fodder beets (Beta vulgaris L.) was studied with four anti-microtubule herbicides: amiprophos-methyl (APM), oryzalin, pronamide, and trifluralin at concentrations of 0–300 μM. Best chromosome doubling results were obtained by treatment of the ovules with 100 μM APM which produced 4.7 diploid plants per 100 ovules. Highest chromosome doubling was found with oryzalin using 1 μM, with trifluralin at 10 μM, and with pronamide at 10 μM producing 2.8, 2.0, and 2.0 diploid plants per 100 ovules, respectively. The APM treatments showed relatively low toxicity on embryo formation which in combination with a high chromosome doubling effect, resulted in up to 89 diploids per 100 plants regenerated. Oryzalin and trifluralin had more severe toxic effects, which reduced embryo formation, thereby lower percentages of chromosome doubled plants were obtained from these treatments. Pronamide had no significant toxic effect but it induced chromosome doubling at lower frequencies. Compared to colchicine, APM seems to be as efficient for chromosome doubling during beet ovule culture, but at molar concentrations 100 times lower than those used for chromosome doubling with colchicine. This revised version was published online in July 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.