Effect of three anthocyaninless genes on germination in tomato (Lycopersicon esculentum Mill.) I. Seed germination under optimal conditions

The effect of three mutant genes affecting anthocyanin biosynthesis – anthocyaninless of Hoffmann (ah), anthocyaninwithout (aw) and baby lea syndrome (bls) – on tomato germination capacities at 24 ° C was investigated. The study was performed on nine anthocyanin containing lines – Apedice, Apeca, Ailsa Craig, Monfavet 167, Monfavet 168, Por, Piernita, VF 36 and VFNT cherry and 13 isogenic/near isogenic lines (IL/NIL) from them: 4 differing for gene ah, 3 differing for gene aw and 6 differing for gene bls). In the majority of the anthocyaninless IL/NILs germination began earlier than that in the wild type lines. Significant differences were observed in the time to 50% germination between the anthocyanin-containing and the anthocyaninless IL/NILs except for the most rapidly germinating line VFNT cherry. Seed weight, water uptake, effect of testa removal and presence or absence of maternal effects in F1 germination responses were investigated in order to elucidate the causes of the more rapid germination in the anthocyaninless IL/NILs. The increased amount of water uptake by the anthocyaninless lines, the shorter treatment time necessary for their testa removal, the important maternal effects in the genetic variability of the time to 50% germination and the differences in germination capacities between the wild type and the mutant IL/NILs after testa removal, indicated that the seed coat and the endosperm had to be affected by the effects of the three genes in a way that caused enhanced germination. This revised version was published online in July 2006 with corrections to the Cover Date.     

Anthocyanin mutations improving tomato and pepper tolerance to adverse climatic conditions

The effect of four mutated genes affecting anthocyanin biosynthesis was investigated relative to tomato and pepper germination capacities and early seedlings and plantlets growth. The study was performed on isogenic/near isogenic lines (IL/NIL) differing for genes ah (Hoffmann’s anthocyaninless), aw (anthocyanin without) and bls (babylea syndrome) in tomato and al ₁ (anthocyaninless – 1) in pepper. Germination responses of the IL/NILs showed that genes ah, aw and bls in tomato and al ₁ in pepper enhanced germination abilities under stress conditions regardless of the inherent seed properties that imparted rapid germination. This evaluation was conducted with four very different types of stress: 13 ^°C, 33 ^°C,120 mM NaCl and 15% PEG-6000 water solutions. The four genes exercised no effect on root and hypocotyl elongation under the same stress conditions and their effect on plantlets growth varied depending on the genotype and the treatment. The usefulness of genes ah, aw, blsand al ₁ in breeding tomato and pepper cultivars tolerant to adverse climatic conditions at germination when used as a morphological marker, is discussed. This revised version was published online in August 2006 with corrections to the Cover Date.     

Identification of near-isogenic lines: an innovative approach,validated for root and shoot morphological characters in a mapping population of rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.)

Near-isogenic lines (NILs) constitute valuable tools in genetic investigations and plant breeding programs. Conventional methods for developing these are time consuming and tedious. An innovative method for identifying NILs is proposed and validated. The method involves computation of simple correlation coefficients of all possible pairs of genotypes within a mapping population using molecular marker data, and phenotypic characterization of those pairs with very high positive correlation. The pairs having both genomic and phenotypic similarity except for a single trait are considered as NILs. This strategy was tested with a doubled haploid mapping population involving CT9993 and IR62266. This population was saturated with 315 markers and comprised 154 lines. The pairs showing very high correlation coefficients (0.70–0.97) and differing for less than 10% of the markers were considered as Genotypically Closely Related Pairs (GCRPs). Graphical genotyping was employed to visualize the genome of the closely related lines. A total of 39 such pairs were subjected to rigorous evaluation for root and shoot morphological traits in two contrasting moisture regimes. Four GCRPs under well-watered condition and ten GCRPs under low moisture stress condition are statistically significant for a single phenotypic trait and are considered as NILs for their respective traits and would be the valuable materials for genetic studies. Mapped QTLs and candidate genes were employed to explain the probable cause of phenotypic difference in NILs.     

Relationships between cold- and salt-tolerance during seed germination in tomato: Germplasm evaluation

Thirty tomato accessions representing six Lycopersicon species were evaluated for the rate of seed germination under no stress, cold-stress and salt-stress conditions. Most accessions responded similarly to both cold- and salt-stress conditions (i.e. they were equally sensitive or tolerant to both stresses), however, a few accessions exhibited more sensitivity (or tolerance) to one stress than the other. In addition, some accessions that germinated relatively rapidly under non-stress conditions exhibited great sensitivity to both cold stress and salt stress. Across accessions, significant (P < 0.01) positive phenotypic correlations were observed between germination rate under control and cold stress (r_P= 0.89), control and salt stress (r_P= 0.63) and cold stress and salt stress (r_P= 0.77). The results indicate that the rate of tomato seed germination under non-stress, cold- and salt-stress conditions may be controlled by the same genes (or physiological mechanisms), but additional components may be involved which affect germination rate under specific stress conditions.     

Comparison of QTLs for seed germination under non-stress, cold stress and salt stress in tomato

The purpose of this study was to examine whether rate of tomato seed germination under non-stress, cold-stress and salt-stress conditions was under similar genetic control by identifying and comparing quantitative trait loci (QTLs) which affect germination rate under these conditions. A fast-germinating accession (LA722) of the wild tomato species Lycopersicon pimpinellifolium Jusl. and a slow-germinating cultivar (NC84173, maternal and recurrent parent) of tomato (Lycopersicon esculentum Mill.) were hybridized and BC₁ and BC₁S₁ progeny produced. The BC₁ population was used to construct a linkage map with 151 restriction fragment length polymorphism (RFLP) markers. The BC₁S₁ population (consisting of 119 BC₁S₁ families) was evaluated for germination under non-stress (control), cold-stress and salt-stress conditions and the mean time to 50% germination (T50) in each treatment was determined. Germination analyses indicated the presence of significant (P < 0.01) phenotypic correlations between T50 under control and cold stress (r = 0.71), control and salt stress (r = 0.58) and cold stress and salt stress (r = 0.67). The QTL analysis indicated the presence of genetic relationships between germination under these three conditions: a few QTLs were identified which commonly affected germination under both stress- (cold-, salt- or both) and non-stress conditions, and thus were called stress-nonspecific QTLs. A few QTLs were also identified which affected germination only under cold or salt stress and thus were called stress-specific QTLs. However, the stress-nonspecific QTLs generally exhibited larger individual effects and together accounted for a greater portion of the total phenotypic variation under each condition than the stress-specific QTLs. Whether the effects of stress-nonspecific QTLs were due to pleiotropic effects of the same genes, physical linkage of different genes, or a combination of both could not be determined in this study. The results, however, indicate that the rate of tomato seed germination under different stress and nonstress conditions is partly under the same genetic control.     

Relationships between cold- and salt-tolerance during seed germination in tomato: Analysis of response and correlated response to selection

Seeds of F₂ progeny of a cross between a slow-germinating (UCT5) and a fast-germinating tomato line (PI120256) were evaluated for germination under non-stress (control), cold-stress and salt-stress conditions, and in each treatment the most rapidly (first 5%) germinating seeds were selected, grown to maturity and self-pollinated to produce F₃ progeny. The selected F₃ progeny from each experiment were evaluated for germination in each of the three treatments, and compared with germination rate of unselected F₃ progeny. Selection for rapid seed germination was effective under cold stress and salt stress, but not effective under non-stress conditions. Furthermore, selection in either cold-stress or salt-stress treatment significantly improved progeny germination rate under both cold-stress and salt-stress treatments, as well as the non-stress treatment. The results support the suggestion that the same genes contribute to rapid seed germination under cold-, salt- and non-stress conditions. In practice therefore, selection for rapid seed germination under a single-stress environment may result in progeny with improved seed germination under a wide range of environmental conditions. Furthermore, to improve germination rate under non-stress conditions, it may be more effective to make selections under a stress treatment.     

旗叶蜡质含量不同小麦近等基因系的抗旱性

于2013-2014和2014-2015年度,以多蜡质和少蜡质的4个小麦近等基因系为材料,采用田间旱棚方式控制土壤水分,研究了蜡质含量与小麦抗旱性的关系。结果表明,干旱处理后,多蜡质小麦品系旗叶的蜡质含量平均为15.15 mg g?1,较少蜡质小麦品系(8.43 mg g?1)高79.8%;多蜡质小麦品系旗叶的水势较高,干旱处理后下降幅度明显小于少蜡质小麦品系,水分散失率也显著低于少蜡质品系(P< 0.05);多蜡质小麦品系旗叶的光合速率平均下降7.5%,而少蜡质小麦品系下降9.8%;多蜡质小麦品系旗叶PSII最大光化学效率(Fv/Fm)平均下降幅度为3.4%,少蜡质小麦品系下降幅度达到5.8%;多蜡质小麦品系的籽粒产量高于少蜡质品系,平均高3.7%;多蜡质小麦品系的抗旱指数和干旱敏感指数均显著低于少蜡质小麦品系(P< 0.05)。以上结果表明,蜡质能够提高小麦的抗旱性,旗叶蜡质含量可以作为抗旱小麦品种的选择指标。     

Absence of a genetic relationship between salt tolerance during seed germination and vegetative growth in tomato

Two approaches were used to determine the relationship between salt tolerance during seed germination and vegetative growth in tomato. First, F₄ progeny families of a cross between a breeding line, ‘UCT5’ (salt sensitive at all developmental stages), and a primitive cultivar, ‘PI 174263’ (salt tolerant during germination and vegetative growth), were evaluated in separate experiments for salt tolerance during germination and vegetative growth. There were significant differences among the F₄ families in both the rate of seed germination and the plant growth (dry matter production) under salt stress. There was, however, no significant correlation between the ability of seeds to germinate rapidly and the ability of plants to grow under salt stress. In the second approach, selection was made for rapid germination under salt stress in an F₂ population of the same cross and the selected progeny was evaluated for salt tolerance during both germination and vegetative growth. The results indicated that selection for salt tolerance during germination significantly improved germination performance under salt stress; a realized heritability estimate of 0.73 was obtained. Selection for salt tolerance during germination, however, did not affect plant salt tolerance during vegetative growth; there was no significant difference between the selected and unselected progeny based on either absolute or relative growth under salt stress. Obviously, in these genetic materials, salt tolerance during germination and vegetative growth are controlled by different mechanisms. Thus, to develop tomato cultivars with improved salt tolerance, selection protocols that include all critical developmental stages would be desirable.     

Genetic analysis of low-temperature tolerance during germination in tomato, Lycopersicon esculentum Mill.

The genetic basis of low-temperature tolerance during germination of tomato seed was investigated using two approaches. First, a cold-tolerant (PI 120256) and a cold-sensitive tomato cultivar (UCT5) and their reciprocal F₂, F₃ and BC₁ progeny (total of 10 generations) were evaluated for germination at a low (11 ± 0.5°C) and a high (control) temperature 20 ±0.5° C) Weighted least-square regression analysis indicated that in the low-temperature treatment most of the variation resulted from additive genetic effects, and dominance and epistatic interactions were nonsignificant. Partitioning of the total genetic variance into those attributable to the effects of embryo, endosperm, testa and the cytoplasm indicated that additive effects of endosperm and embryo could individually account for 80% and 77% of the total variance, respectively. In the control treatment, greater than 60% of the variation could be explained by individual additive effects of endosperm or embryo and ? 27% of the variation could be explained by embryo dominance effects. Across generations, there was a positive correlation (r = 0.78, P < 0.01) between germination in the control and low-temperature treatments and there were no significant genotype × temperature interactions. The results indicate the presence of similar or identical genes with predominantly additive effects on germination under both low and high temperatures. In the second approach, the effectiveness of directional phenotypic selection to improve tomato cold tolerance during germination was evaluated by selecting (in an F₂ population of the same cross) the fastest germinating seeds under low temperature and comparing the germination of the selected F₃ progeny with germination of an unselected F₃ population. The results indicated that selection was highly effective and significantly improved germination performance of the progeny; a realized heritability of 0.74 was obtained for low-temperature tolerance during germination. It is concluded that in these tomato lines germination under low temperature is genetically controlled, with additivity being the major genetic component, and thus the trait can be improved by phenotypic selection.     

Genetic relationships among cold, salt and drought tolerance during seed germination in an interspecific cross of tomato

Seed of BC₁ progeny of an interspecific cross between a slow germinating Lycopersicon esculentum breeding line(NC84173; maternal and recurrent parent) and a fast germinating L.pimpinellifolium accession (LA722) were evaluated for germination under cold stress, salt stress and drought stress, and in each treatment the most rapidly germinating seeds (first 2%) were selected. Selected individuals were grown to maturity and self-pollinated to produce BC₁S₁ progeny families. The selected BC₁S₁ progeny from each experiment were evaluated for germination rate in each of a non stress (control),cold-, salt- and drought-stress treatment, and their performances were compared with those of a non selected BC₁S₁population in the same treatments. Results indicated that selection for rapid seed germination in each of the three stress treatments was effective and significantly improved progeny germination rate under all three stress conditions. The results support the suggestion that same genes might control the rate of seed germination under cold, salt and drought stress. Furthermore, selection in each of the three stress treatments resulted in improved progeny seed germination rate under nonstress conditions, suggesting that genetic mechanisms that facilitate rapid seed germination under stress conditions might also contribute to rapid germination under nonstress conditions. In practice, therefore, selection for rapid seed germination under a single stress environment may result in progeny with improved seed germination under a wide range of environmental conditions. Furthermore, to improve germination rate under nonstress conditions, it might be more efficient to make selections under stress conditions. This revised version was published online in July 2006 with corrections to the Cover Date.     

Combining ability of low phytic acid (<Emphasis Type="Italic">lpa1</Emphasis>-<Emphasis Type="Italic">1</Emphasis>) and quality protein maize (QPM) lines for seed germination and vigour under stress and non-stress conditions

Grain yield is more likely to be compromised by poor seed germination and vigour in low phytic acid (LPA) and quality protein maize (QPM) than normal maize (Nm), especially when grown under stressful tropical environmental conditions. The objectives of this study were to determine the effect of stress conditions on seed germination and vigour traits (percentage germination, seedling dry weight, average root and shoot length and vigour index) and to determine the GCA effects of the parental lines and SCA effects of the crosses. A ten parent half diallel (LPA, Nm, QPM lines) was subjected to the standard germination (non-stress) and accelerated aging (AA) (stress) tests. All seed lots were at the same physiological age and produced under the same season and conditions. Genotypic and group differences were investigated. General combining ability (GCA) and specific combining ability (SCA) effects were significant (P ≤ 0.001) for all traits under both stress and non-stress conditions indicating that both additive and non-additive gene effects were significant. Generally SCA effects were superior for all traits. The LPA lines displayed 61 % reduction in germination and 23–52 % reduction in vigour under stress conditions thereby underscoring challenges that are expected in deploying LPA maize in stress conditions. However, LPA line CM 31 exhibited large positive and mostly significant GCA effects, while two LPA × LPA crosses showed significant positive SCA effects. Results indicate breeding is required to improve both germination and vigour of the LPA lines to adapt them to tropical conditions that are generally stress-prone.     

干旱条件下冬小麦近等基因系水分利用效率及其与气冠温差的关系

鉴定比较供体亲本京411和轮回亲本晋麦47及其34个近等基因系材料之间产量和水分利用效率(WUE),同时测定不同生育期的气冠温差(CTD),分析WUE在这些材料之间的变异范围,筛选WUE与轮回亲本显著差异的材料用于QTL定位,同时探索干旱条件下CTD与产量和WUE的关系及其随生育期进程变化的趋势。利用防雨棚和渗漏池开展模拟干旱试验。结果表明,34个近等基因系及其父母本WUE为1.30~1.92 kg/m~3,其中9个品系WUE显著低于轮回亲本晋麦47,表明WUE为多基因控制的数量性状,而且存在明显的加性效应。CTD与产量和WUE均呈极显著正相关关系。不同的是,随生育期进程推进,CTD与产量的相关性增强,在灌浆期最高,R2达到0.684 9,而与WUE的相关性减弱,R2在拔节-孕穗期最高,达到0.769 8。研究初步表明,CTD可以作为干旱条件下产量和WUE的鉴定指标。     

Effects of high oleic acid soybean on seed yield,protein and oil contents,and seed germination revealed by near‐isogeneic lines

Typical soybean oil is composed of palmitic, stearic, oleic, linoleic and linolenic acids. High oleic acid content in soybean seed is a key compositional trait that improves oxidative stability and increases oil functionality and shelf life. Using a marker‐assisted selection method, near‐isogenic lines (NILs) of G00‐3213 for the high oleic trait were developed and yield tested. These NILs have various combinations of FAD2‐1A and FAD2‐1B alleles that were derived from the same backcrossing populations. The results indicated that G00‐3213 NILs with both homozygous mutant FAD2‐1A and FAD2‐1B alleles produced an average of 788 g/kg oleic acid content. The results also demonstrated that possessing these mutant alleles did not cause a yield reduction. Furthermore, seed germination tests across 12 temperatures (12.8–32.0°C) showed that modified seed composition for oleic acid in general did not have a major impact on seed germination. However, there was a possible reduction in seed germination vigour when high oleic seeds are planted in cold soil. The mutant FAD2‐1A and FAD2‐1B alleles did not hinder either seed or plant development.     

In vitro pollen germination and tube growth of tomato (Lycopersicon esculentum Mill.) and its relation with plant growth

Summary In vitro pollen germination at 10°C, 14°C and 22°C of four groups of two pure line tomato varieties was compared with their plant growth at 19°C D/10°C N under controlled environmental conditions. Generally, pollen germination was slow at 10°C but after 6 h the percentage of germination was similar to that at 22°C. Maximum germination was obtained at 14°C already after 4 h. The longest pollen tubes occurred at 22°C. The two varieties within each group differed significantly from each other for percentage of pollen germination. For one group, this difference was greater at 10°C than at 22°C. The varieties in two groups also differed significantly for pollen tube growth. These differences in pollen tube growth rate were greater at 22°C than at 10°C. There was no clear relationship between pollen germination, pollen tube growth and plant growth in any of the four pairs of varieties. The results are discussed with regard to the possibility of pollen selection at low temperature in order to improve the efficiency of breeding for growth at low temperature.     

Tagging the juvenile locus in soybean [Glycine max (L.) Merr.] with molecular markers

Soybean cultivars carrying the `long juvenile trait' show a delayed flowering response under short day conditions. The incorporation of this character into genotypes of agronomic interest may allow a broader range of sowing dates and latitudes for a single cultivar adaptation. The objective of this work was to identify molecular markers linked to the juvenile locus in soybean. Experiments were carried out using two pairs of near isogenic lines(NILs) differing in the presence of the long juvenile trait, and RAPD markers. Four hundred primers were first screened to find polymorphism associated with the trait. Additional differences between NILs were sought by digesting the genomic DNA with five restriction enzymes. Polymorphic fragments detected between NILs were tested for linkage to the juvenile locus in the corresponding F₂ segregating populations. Marker bc357-HaeIII was linked (χ²L = 46.316) to the juvenile locus with an estimated recombination frequency of 0.13 ± 0.03in one of the genetic backgrounds studied. The fragment was cloned, sequenced and converted into a SCAR marker. Moreover,bc357-HaeIII was used as RFLP probe. Both, SCAR and RFLP generated markers linked to the juvenile locus in the two genetic backgrounds analysed. Results presented in this work can be utilised for both, the localisation of the gene associated with the character and for tagging the juvenile trait in soybean breeding programs. This revised version was published online in August 2006 with corrections to the Cover Date.     

Development of NILs from heterogeneous inbred families for validating the rust resistance QTL in peanut (Arachis hypogaea L.)

Heterogeneous inbred families segregating for rust resistance were identified from the two crosses involving susceptible (TAG 24 and TG 26) and resistant (GPBD 4) varieties of peanut. Rust‐resistant (less than score 5) and rust‐susceptible (more than score 5) plants were identified in each HIF and evaluated under rust epiphytotic conditions. The set of plants belonging to the same HIF, but differing significantly in rust resistance, not in other morphological and productivity traits, was regarded as near‐isogenic lines (NILs). Largely, rust‐resistant NILs had GPBD 4‐type allele, and susceptible NILs carried either TAG 24 or TG 26‐type allele at the three SSR loci (IPAHM103, GM1536 and GM2301) linked to a major genomic region governing rust resistance. Comparison of the remaining genomic regions between the NILs originating from each of the HIFs using transposon markers indicated a considerably high similarity of 86.4% and 83.1% in TAG 24 × GPPBD 4 and TG 26 × GPBD 4, respectively. These NILs are useful for fine mapping and expression analysis of rust resistance.     

Effect of row type,flowering type and several other spike characters on resistance to Fusarium head blight in barley

Fusarium head blight (FHB) is a destructive disease of barley. The genetics and expression of resistance to FHB in barley is complex, and various spike characters are thought to possibly influence resistance. Tests using spray-inoculation of Fusarium graminearum at anthesis in greenhouse environments showed that two-rowed and cleistogamous varieties from Japan belong to the highest resistance group, while six-rowed and chasmogamous varieties are mostly susceptible. In order to evaluate the effect of such spike characters, including row type and flowering type, on FHB resistance, near-isogenic lines (NILs) differing in these characters were tested for their resistance. Two testing methods were used: the pot-plant and cut-spike methods, in which spikes at anthesis were spray-inoculated in greenhouse environments. The chasmogamous NILs and some six-rowed NILs were significantly more diseased than cleistogamous and two-rowed parent lines, respectively, and the difference in FHB severity was greater and more stable between cleistogamous/chasmogamous NIL pairs than between two-/six-rowed pairs. Slight or no differences were observed in glaucous/non-glaucous, normal/dense spike, normal/uzu type and normal/deficiens NIL pairs. The results indicate that the contribution of cleistogamy and/or the genetic background toward FHB resistance is more than that of row type and the other tested spike characters. Further, it should be possible to develop six-rowed varieties with FHB resistance nearly as good as that of the two-rowed varieties.     

Genetic and non-genetic factors affecting germination and vitality in spring barley seed

Germination of eight spring barley varieties, grown in two years at eight different locations in State varietal trials in the Czech republic, was analysed. All the combinations were tested under optimal conditions for germination as recommended by ISTA and also at low temperatures (5 and 10°C), and underwater stress (- 2 and - 4 bar). Germination under low temperature and water stress was considered as a character of seed vitality. Varieties in nearly all experiments were a significant source of variation and their relative contribution to the total variation was higher for vitality (5.7–38.7%) than for germination (2.0–4.3%). However, the percentage of germination were not related to the percentage of vitality. The location in nearly all the experiments caused the greatest variation and the impact was greater for germination than for vitality. Provenance effects were different for germination and for vitality; grains harvested in the lowland had the highest germination values, but not the highest vitality values. The results should encourage breeding for seed vitality and some reconsideration of provenance effects on seed vitality.     

转ER-sHSP基因番茄植株的耐盐性检测

热激蛋白在植物应对各种环境胁迫的应答中执行重要的功能并存在交叉保护现象。前期结果表明内质网小分子热激蛋白（ER-sHSP）的过量表达能够显著增强转基因番茄的抗冷能力,为继续深入研究ER-sHSP的生理功能,本文对高盐胁迫下转ER-sHSP基因番茄的耐受性进行了检测,结果显示：与对照相比,转基因番茄抗性表型明显,持有较高的植物干鲜重,脯氨酸含量大幅度提高,SOD酶活性增加明显,而MDA含量和电导率则较低。说明ER-sHSP的过量表达提高了转基因番茄的抗盐胁迫能力     

Development of near-isogenic lines for a major QTL on 3BL conferring Fusarium crown rot resistance in hexaploid wheat

By essentially fixing the genetic background, near-isogenic lines (NILs) are ideal for studies of the function of specific loci. We report in this paper the development of NILs for a major QTL located on the long arm of chromosome 3B conferring Fusarium crown rot (FCR) resistance in hexaploid wheat. These NILs were generated based on the method of the heterogeneous inbred family analysis. 13 heterozygous lines were initially selected from three segregating populations using a single SSR marker linked with the major FCR QTL. The two isolines for each of the putative NILs obtained showed no obvious morphological differences, but differences among the NIL pairs were large. Significant differences in FCR resistance between the isolines were detected for nine of the 13 putative NIL pairs. The presence of the FCR allele from the resistant parent reduced FCR severity by 29.3–63.9% with an average of 45.2% across these NILs. These NILs will be invaluable in further characterising this major FCR locus, in studying the mechanism of FCR resistance and in investigating possible interactions between FCR resistance and other traits of agronomic importance.