Novel variation for the tocopherol profile in a sunflower created by mutagenesis and recombination

α‐Tocopherol is the main tocopherol in sunflower seeds (>90%). Because it exerts a weak antioxidant action in vitro, its partial replacement by other tocopherols is an important breeding objective in this crop. The objective of this research was to develop novel tocopherol profiles in sunflower through mutagenesis and genetic recombination. Seeds of four ‘Peredovik’ accessions were used for chemical mutagenesis with ethyl methane sulfonate (EMS). Single‐seed screening in the M₂ generation resulted in two M₂ seeds, derived from different M₁ plants, with increased γ‐tocopherol contents of 19.2% and 96.7%, respectively. M₃ progeny from the M₂ seed with the 96.7% content bred true for high c‐tocopherol content, containing more than 90%γ‐tocopherol. M₃ progeny from the M₂ seed with only 19.2%γ‐tocopherol segregated in a range from 0 to 84.6%. Selection for high c‐tocopherol content produced an M_4:5 line, designated IAST‐1, with a stable high concentration of γ‐tocopherol. Crosses between IAST‐1 and T589, with an increased b‐tocopherol content, produced F₂ segregants with trans‐gressive levels of up to 77%β‐tocopherol or up to 68% d‐tocopherol. Both novel tocopherol profiles were confirmed in the F₃ generation.     

Gamma Ray-Induced Field Tolerance to Phytophthora Blight in Sesame

Due to the non-availability of sources of resistance in Sri Lanka to Phytopbthora blight in sesame (Sesamum indicum) caused by Phytopbthora nicotianae var. parasitica, a mutation breeding programme was initiated. Seeds of three genotypes were subjected to six doses of gamma rays from 100 Gy to 750 Gy from a ⁶⁰Co source. Seeds of M₂ bulks, sampled by variety and treatment from the first five capsules formed on M₁ plants, were grown in a field with a history of repeated incidence of the disease. The best 21 lines, having the highest survival and seed production, identified in the screening of M₃ and M₄ progeny rows of selected M₂ single plants were then tested in a replicated field trial with the recommended cultivar ‘MI 3’ as a control. The plant survival in selected lines averaged 43.3 percent as against 7.2 percent in the recommended variety. Eight selections recorded significantly higher seed yield than ‘MI 3’ at P < 0.01 and another three at P < 0.05. Gamma ray treatments of 450 Gy and 600 Gy produced more lines tolerant to the disease than the other doses used.     

理化诱变小豆京农6号突变体的鉴定

选用小豆品种京农6号种子,分别采用甲基磺酸乙酯(EMS)(0.5%、0.9%和1.4%处理12h和24h)、电子束(100、300、600Gy)、60Co-γ(400Gy)诱变处理,将处理后的种子种于大田,鉴定后代植株性状的变异。观察表明,EMS诱变的变异类型最丰富、60Co-γ射线次之、电子束产生的变异类型较单一。EMS处理小豆以浓度0.5%和0.9%处理24h为宜;0.5%EMS处理的粒色和荚色变异突出,有鲜红、黄白、绿白粒色和黑荚、褐荚、黑褐荚变异;0.9%处理的叶形变异突出,有鸡爪叶、剑叶、肾形叶、小密叶等突变类型;电子束诱变后,M2变异率分别为4.09%、3.64%和2.22%。400Gy60Co-γ射线处理种子,后代变异率为7.23%。通过两年的鉴定筛选,获得937个EMS诱变M3代株系,934个60Co-γ射线和电子束诱变M2代株系,已得到株高、叶形、叶色、粒形、粒色、荚色、无分枝、多分枝、叶簇生、分枝簇生、光叶、蔓生、有限结荚习性、株型松散、育性、成熟特性等突变体材料1490份。本研究为小豆基因遗传分析、基因定位与克隆及其进一步的基因功能分析奠定了基础,为小豆育种提供了重要的材料。     

Variability among advanced gamma-irradiation induced large-seeded mutant breeding lines in the 'Georgia Browne' peanut cultivar

The objective of this study was to compare the variability among advanced large‐seeded ‘Georgia Browne’ mutant breeding lines induced by γ‐irradiation. Seeds of the small‐seeded, high‐yielding, disease‐resistant peanut cultivar ‘Georgia Browne’ were exposed to a 200 Gy dose of γ‐radiation. Several advanced (M_6:7–M_6:9)‘Georgia Browne’ mutant breeding lines were developed and evaluated at the University of Georgia, Coastal Plain Experiment Station over three consecutive years 1997‐99 for disease incidence, pod yield, total sound mature kernels (TSMK) grade, pod weight, seed weight and seed size distribution. Field performance tests showed significant differences among the advanced large‐seeded mutant breeding lines compared with ‘Georgia Browne’ for each of these variables. The results obtained demonstrate the beneficial use of mutation breeding for inducing and developing variable and desirable advanced mutant breeding lines within peanut cultivars.     

Breeding for Modified Fatty Acid Composition by Induced Mutations in Linseed (Linum usitatissimum L.)

Seeds of linseed (Linum usitatissimum L.) cultivars ‘Raulinus’ and ‘Bionda’ were treated mutagenically with ethyl methanesulfonate (EMS) in order to broaden the variation of fatty acid composition in the seed oil and to select mutants with reduced linolenic acid (C18: 3) content. The M₂ generation was evaluated for fatty acid composition of the oil. Genetic variation for unsaturated fatty acids was demonstrated to be extended in the M₂ generation as compared to the controls. Finally in the M₅ generation of cv. ‘Raulinus’, lines with reduced C18:3 (< 40% vs. 55%) and correspondingly increased linoleic or oleic acid contents, respectively, could be selected. These mutants proved to be genetically stable as demonstrated by parent-offspring (M₄/M₅) regression.     

不同诱变处理对苜蓿叶绿素含量及光合作用的影响

为研究60Co-γ射线、紫外线、零磁空间和EMS诱变处理对苜蓿SP1代叶片叶绿素含量及光合作用的影响,筛选最优诱变处理梯度,选育适合高寒地区种植的苜蓿新品系或品种。以‘公农1 号’、‘Wega7F’、‘WL319HQ’、‘敖汉’4 个苜蓿品种试验材料,设置60Co-γ射线150、300、450 Gy射线强度,紫外线30、60、90 min 照射时间,EMS 0.1%、0.2%、0.4%(v/v)溶液浓度,零磁空间处理180 天;测定苜蓿叶片叶绿素a、叶绿素b、叶绿素总量、叶绿素a/b 值以及净光合速率。结果表明,‘公农1 号’450 Gy60Co-γ处理、紫外线处理时间为30 min和0.4%EMS处理下叶绿素a 含量最高,均高于对照3.17 mg/g,但与对照差异不显著,其他处理均低于对照。‘Wega7F’在紫外线60 min 时间处理下,叶绿素b 值明显增加,且显著高于其他处理(P<0.05)。‘公农1 号’在450 Gy 60Co-γ辐射处理下叶绿素含量最高,a/b 比值最小,其他3 个品种在150 Gy处理下叶绿素含量最高,但a/b 比值最小。各苜蓿品种不同诱变处理净光合速率大小依次为：60 min 紫外线处理的‘Wega7F’>零磁空间处理的‘敖汉’>0.4%EMS 处理的‘WL319HQ’>0.4%EMS处理下的‘公农1 号’。150 Gy60Co-γ射线、60 min 紫外线、0.4%浓度EMS处理为苜蓿正向变异处理,有利于筛选有益诱变突变体。     

Interaction Amongst Soybean [Glycine max (L.) Merrill] Genotype, Soil Type and Inoculant Strain with Regard to N2 Fixation

The nitrogen (N₂)‐fixing bacterial inoculant strain for soybean [Glycine max (L.) Merrill] is not indigenous to South African soils. The interaction between soybean genotype, soil type and inoculant strain, however, has a definite influence on soybean production and compatibility should be optimized. This paper reports a growth chamber study using three different soybean genotypes (Barc‐9, Avuturda and Talana), three Bradyrhizobium japonicum inoculant strains (WB108, WB112 and WB1) and three soil types (Avalon, Arcadia and sand) to evaluate the effectiveness of N₂ fixation by different genotype × soil type × inoculant strain combinations, using different measuring parameters. These parameters included nodule fresh mass (NFM), amount of N₂ fixed (P_fix), as determined by the ureide method, seed protein content (SPC), average seed mass per plant (SMP) and average foliar N content (FNC). The comparison amongst the three‐way interactions, genotype × soil type × inoculant strain, did not differ significantly for the parameters used. Significant two‐way interactions were soil × inoculant for FNC, P_fix and SMP; soil × genotype for FNC and SMP, and inoculant × genotype for FNC (P < 0.05). The soil × inoculant strain interaction was significant for P_fix (P < 0.05). NFM, P_fix, FNC, SMP and SPC correlated positively with soil pH and negatively with soil clay content and soil NO₃^– and NH₄⁺ content (P < 0.05). SPC was significantly different (P < 0.05) for soil type, genotype and inoculant strain. P_fix and NFM did not reflect the protein content of the seeds, indicating that nodule evaluation should be used with caution as a N₂ fixation parameter. Low soil pH and high mineral N content inhibited N₂ fixation. NFM correlated negatively with the clay content of the soil. This finding confirms that soybean production in South Africa can be improved by appropriate selection of genotypes and inoculant strains for their compatibility in different soils.     

Screening of Chinese cabbage mutants produced by 60Co γ‐ray mutagenesis of isolated microspore cultures

Since the release of the Chinese cabbage genome sequence, increasing interest has focused on the functional analysis of unidentified genes in Chinese cabbage. Mutant analysis forms the basis of functional genomics research. To produce a variety of Chinese cabbage mutants in the same genetic background, buds containing late uninucleate spores from a doubled haploid line of the Chinese cabbage variety ‘Fukuda 50’ were irradiated with ⁶⁰Co γ‐rays at doses of 20, 40 and 60 Gy. Then, the treated microspores were isolated and cultured. A total of 492 putative M₀ mutants were isolated from 1483 regenerated plants. Of these, six M₁ mutants were verified; the mutant frequency was 0.41%. These mutants comprise a mutant library that includes one plant shape mutant, two flower mutants and three male sterile mutants. Pollen viability detection and DNA flow cytometry were used to determine the ploidy of the regenerated plants. Some of the mutants isolated in this study may be useful for Chinese cabbage breeding and functional genomics research.     

大豆“南农86-4”突变体筛选及突变体库的构建

应用⁶⁰Co γ射线和甲基磺酸乙酯(EMS)分别对“南农86-4”大豆种子进行诱变，并构建大豆突变体库。结果在M₃代分别获得40份和145份叶、茎、花、种子、子叶等性状变异的材料。在两种方式诱变的M₃代都获得蛋白质和油含量变化的材料，尤其在EMS诱变的M₃代中获得47份蛋白质含量比对照高5个百分点以上的材料，5份蛋白质和油总含量比对照高5个百分点以上的材料。这些突变体可以作为新的种质资源，构建的突变体库也有助于大豆功能基因组研究的开展。     

Quantitative trait loci underlying soybean seed tocopherol content with main additive,epistatic and QTL × environment effects

Soybean (Glycine max [L.] Merrill) seeds are a major source of tocopherols (Toc), which could significantly improve immune system health of human and prevent or treat many serious diseases. Selection for higher Toc contents of seeds could increase nutritional value of soybean‐derived food, laying on an important breeding goal for many soybean breeders. The present objectives of the work were to evaluate various genetic effects of QTL associated with individual and total Toc content based on a RIL population (“Beifeng 9” × “Freeborn”) in six environments to improve the efficiency of molecular marker‐assisted selection (MAS) for high‐Toc breeding. The results described that eighteen, thirteen, eleven and thirteen QTL were associated with α‐Toc, γ‐Toc, δ‐Toc and total Toc content, respectively, and have additive main effects (a) and/or additive × environment interaction effects (ae) in certain environments. Among them, four QTL for α‐Toc, two QTL for γ‐Toc, one QTL for δ‐Toc and four QTL for total Toc could increase α‐Toc, γ‐Toc, δ‐Toc and total Toc content via significant a effect, respectively, which have stronger stability in different years and locations. It implied a value for MAS. Additionally, twenty‐five, fifteen, eleven and twenty epistatic pairwise QTL associated with α‐Toc, γ‐Toc, δ‐Toc and total Toc contents, respectively, were detected. The genetic information of the QTL effects obtained here would be beneficial for breeding soybean variety with high‐Toc content by MAS.     

Characterization of broad‐spectrum resistance to Soybean mosaic virus in soybean [Glycine max (L.) Merr.] cultivar ‘RN‐9’

Soybean mosaic virus (SMV) can cause serious yield losses in soybean. Soybean cultivar ‘RN‐9’ is resistant to 15 of 21 SMV strains. To well‐characterize this invaluable broad‐spectrum SMV‐resistance, populations (F₁, F₂ and F_2:3) derived from resistant (R) × susceptible (S) and R × R crosses were tested for SMV‐SC18 resistance. Genetic analysis revealed that SC18 resistance in ‘RN‐9’ plus two elite SMV‐resistant genotypes (‘Qihuang No.1’ and ‘Kefeng No.1’) are controlled by independently single dominant genes. Linkage analysis showed that the resistance of ‘RN‐9’ to SMV strains SC10, SC14, SC15 and SC18 is controlled by more than one gene(s). Moreover, Rsc10‐r and Rsc18‐r were both positioned between the two simple sequence repeats markers Satt286 and Satt277, while Rsc14‐r was fine‐mapped in 136.8‐kb genomic region containing sixteen genes, flanked by BARCSOYSSR_06_0786 and BARCSOYSSR_06_0790 at genetic distances of 3.79 and 4.14 cM, respectively. Allelic sequence comparison showed that Cytochrome P450‐encoding genes (Glyma.06g176000 and Glyma.06g176100) likely confer the resistance to SC14 in ‘RN‐9’. Our results would facilitate the breeding of broad‐spectrum and durable SMV resistance in soybeans.     

Analysis of generation means and components of variance for parthenocarpy in cucumber (Cucumis sativus L.)

Parthenocarpy (seedless fruit) has potential for increasing yield in cucumber (Cucumis sativus var. sativus L.). To determine the inheritance of parthenocarpy in gynoecious cucumber, P₁, P₂, F₁, F₂, BC₁P₁, and BC₁P₂ generations derived from crossing two non‐parthenocarpic gynoecious inbred lines [Gy8 (P₂; processing type) and ‘Marketmore 80’ (P₂; MM, fresh market type)] with a highly parthenocarpic inbred line [2A (P₁; processing type)] were evaluated for fruit number in a greenhouse at Arlington, Wisc. in 1999 (designated 2A × Gy8 1999) and in the open‐field at Hancock, Wisc. in 2000 (designated 2A × Gy8 2000 and 2A × MM 2000). There were significant location and location × generation interaction effects, and therefore generation means analyses were conducted separately for each location. The minimum numbers of effective factors controlling parthenocarpy were estimated to be at least one (2A × Gy8 1999), two (2A × Gy8 2000) and four (2A × MM 2000). Results suggest that selection for parthenocarpy for multiple hand harvest operations will likely be more effective than that for once‐over machine harvest operations. However, the selection efficiency will likely vary across different populations and environments.     

Relationships between different prolamin proteins and some quality properties in durum wheat

The objective of this paper was to study the differences between some prolamin variants coded at the loci Glu-3/Gli-1, Glu-1 and Gli-A2 and their relative roles in durum-wheat quality. F₃ lines from four durum wheat crosses (‘Abadia’בMexicali’. ‘Oscar’בArdente’, ‘Oscar × Mexicali’ and ‘Alaga’בC. of Balazote’) were analysed for gliadin and glutenin composition by electrophoresis. Whole-grain-derived samples were analysed for SDS sedimentation (SDSS) value, mixing properties, and contents of protein and vitreousness. The glutenin patterns LMW-2. LMW-2^? and LMW-2 (CB) at Glu-B3/Gli-B1 were associated with better gluten quality than were LMW-1 and LMW-2*. The glutenin subunits LMW4 and LMW3 + 15 at Glu-A3/Gli-A1 and HMW-1 showed better mixing properties than LMW7 + 12, LMW5 and the null phenotype. respectively. The HMW glutenin subunits 20 + 8 at Glu-B1 showed a negative association with gluten quality, but the rest of the HMW glutenin subunits and α-gliadins did not show any influence on gluten quality. Correlations between the results of the SDSS test and the mixograph were highly significant, but no correlation was found between these results and protein and vitreousness contents. The results are discussed in relation to the development of durum wheat varieties with improved qualities.     

Mapping quantitative trait loci (QTLs) underlying seed vitamin E content in soybean with main,epistatic and QTL × environment effects

Soybean (Glycine max (L.) Merr.) seed contains small amounts of tocopherol, a non‐enzymatic antioxidant known as lipid‐soluble vitamin E (VE). Dietary VE contributes to a decreased risk of chronic diseases in humans and has several beneficial effects on resistance to stress in plants, and increasing VE content is an important breeding goal for increasing the nutritional value of soybean. In this study, quantitative trait loci (QTLs) underlying VE content with main, epistatic and QTL × environment effects were identified in a population of F_5 : 6 recombinant inbred lines from a cross between ‘Hefeng 25’ (a low‐VE cultivar) and ‘OAC Bayfield’ (a high‐VE cultivar). A total of 18 QTLs were detected that showed additive main effects (a) and/or additive × environment interaction effects (ae) in different environments. Moreover, 19 epistatic pairs of QTLs were found to be associated with α‐tocopherol (α‐Toc), γ‐tocopherol (γ‐Toc), δ‐tocopherol (δ‐Toc) and total VE (TE) contents. The QTLs identified in multienvironments could provide more information about QTL by environment interactions and could be useful for the marker‐assistant selection of soybean cultivars with high seed VE contents.     

Identification and validation of an over‐dominant QTL controlling soybean seed weight using populations derived from Glycine max × Glycine soja

Heterosis, or hybrid vigour, has been used to improve seed yield in several important crops for decades and it has potential applications in soybean. The discovery of over‐dominant quantitative trait loci (QTL) underlying yield‐related traits, such as seed weight, will facilitate hybrid soybean breeding via marker‐assisted selection. In this study, F₂ and F_2 : 3 populations derived from the crosses of ‘Jidou 12’ (Glycine max) × ‘ZYD2738’ (Glycine soja) and ‘Jidou 9’ (G. max) × ‘ZYD2738’ were used to identify over‐dominant QTL associated with seed weight. A total of seven QTL were identified. Among them, qSWT_13_1, mapped on chromosome 13 and linked with Satt114, showed an over‐dominant effect in two populations for two successive generations. This over‐dominant effect was further examined by six subpopulations derived from ‘Jidou12’ × ‘ZYD2738’. The seed weight for heterozygous individuals was 1.1‐ to 1.6‐fold higher than that of homozygous individuals among the six validation populations examined in different locations and years. Therefore, qSWT_13_1 may be a useful locus to improve the yield of hybrid soybean and to understand the molecular mechanism of heterosis in soybean.     

AFLP–SSR maps of maize × teosinte and maize × maize: comparison of map length and segregation distortion

Two genetic linkage maps of Zea mays were constructed: one population comprised 94 F₂ individuals of a dent ‘B64’ × teosinte (Z. mays ssp. huehuetenangensis) cross while the second consisted of 94 F₂ individuals of a ‘B64’ × Caribbean flint ‘Na4’ cross. The level of polymorphism was higher in the ‘B64’ × teosinte combination than the ‘B64’ × ‘Na4’ combination. In the ‘B64’ × teosinte cross, a total of 338 amplified fragment length polymorphism (AFLP) and 75 simple sequence repeat (SSR) markers were mapped to 10 chromosomes, which covered 1402.4 cM. In the ‘B64’ × ‘Na4’ cross, a total of 340 AFLP and 97 SSR markers were mapped to 10 chromosomes, covering 1662.8 cM. Segregation distortion regions were found on chromosomes 4, 5 and 8 in the ‘B64’ × teosinte cross and on chromosome 9 in the ‘B64’ × ‘Na4’ cross. Comparison of the two maps revealed that the maize × teosinte map was 11.5% shorter than the maize × maize map. The maps generated in this study may be useful to identify genes controlling flooding tolerance.     

Inheritance of fiber quality and lint yield in a chemically mutated population of cotton

The narrow germplasm base of the upland cotton (Gossypium hirsutum L.), grown on the Texas high plains historically, has limited improvement of fiber quality. Chemical mutagenesis and subsequent selection have helped the development of lines with improved fiber quality in cultivars adapted to this region. This study was conducted to determine the inheritance of improvements in fiber quality. M₃ lines with divergent fiber properties of micronaire, length, and strength were selected from a population of Paymaster HS 200 treated with 3% v/v ethyl methanesulfonate (EMS) for two hours. The 115 selected lines of M₄ and M₅ generation were evaluated for fiber quality and lint yield. Regression of the M₄ and M₅ on the M₃ generation, as well as the M₅ on the M₄ was used to generate narrow sense heritability coefficients. Significant variations were observed between the mutant lines in all generations except for lint yield in the M₅ (1997). The highest heritability estimates were found in fiber length (h ²= 0.29** to 0.46**). Micronaire and strength showed intermediate heritability estimates of h ²= 0.14 to 0.19, while lint yield had a very low heritability estimate of h ²= 0.03. Fiber length and strength were correlated (r= 0.58** to 0.46**) in all the three generations. The mutants identified in these studies have the potential to improve fiber quality of upland cotton without introducing alien genes that may reduce adaptation to short growing season production regions.     

Induced Vine Mutant in Vigna mungo

Pure dry seeds of variety T₉ of Vigna mungo were irradiated by 10, 20 and 30 kR of gamma rays at 33 °C followed by post irradiation treatment with 0.25 % EMS in phosphate buffer (pH 7.0) at 30 ± 1 °C. Vine mutants were isolated from segregating families after 20 kR and 10kR ± EMS treatment. The genetic nature of the mutants was tested by crossing with the wild type. The segregation in F² proved the mutant to be monogenic recessive. The mutant showed slight improvement in protein content and marked improvement in yield of seeds.     

Induced mutations for reduced tuber glycoalkaloid content in potatoes

Mutation breeding was used to produce mutants of the potato breeding selection NDA1725-1 with reduced levels of tuber glycoalkaloids. Excised tuber eye-pieces were exposed to 35 Gy of gamma rays from a ⁶⁰Co source. In the M₁V₁, leaf tissue was screened using high-performance thin layer chromatography and each plant given a relative score based on a subjective rating of band intensity. Tubers were harvested from all plants that produced leaves with relatively low levels of glycoalkaloids. Tuber glycoalkaloids were quantified in the M₁V₂ and M₁V₃ using high-performance liquid chromatography, and 35% and 23% of clones with the lowest glycoalkaloid content were selected for further evaluation in each generation, respectively. In the M₁V₄, M₁V₅ and M₁V₆, selection was based on statistically significant differences between the mutant clones and the controls for glycoalkaloid content, as determined using a gravimetric quantification method, as well as on agronomic and quality characteristics. At the end of six clonal generations three selections exhibited acceptable levels of tuber glycoalkaloids and retained enough of the parent clone's exceptional traits to have potential in the commercial potato chip (crisp) industry.     

Plant Regeneration from in vitro Cultured Anthers of Black Mustard (Brassica nigra Koch)

Anthers of Brassica nigra, excised from fresh as well as cold-pretreated (3 days at 3 ± 2°C) buds cultivated on modified B₅ medium (Gamborg et al. 1968) containing sucrose level varying from 2 % to 10 %, along with 1O^?6M BAP (benzylaminopurine) and 9 × 10^?6M 2,4-D (2,4-dichlorophenoxyacetic acid), developed calli and/or embryos. The latter response was observed only in anthers reared on media containing 6 % or higher levels of sucrose. On media containing two or four per cent sucrose, the anthers produced calli, exclusively. The growth of embryos was inhibited or else they started callusing if left on the media containing higher levels of sucrose. However, on transfer to MS medium (Murashige and Skoog 1962), containing 2 % sucrose, embryos started callusing and subsequently a few secondary embryos differentiated. Such embryos were sub-cultured on MS + 5 × 10^?6M BAP + 2 % sucrose, wherein numerous shoots developed from embryos. The shoots were rooted by transferring to a medium containing 5 × 10^?6M NAA (naphthalene acetic acid). Within two months of culture, some of these plants started flowering in vitro.