不同播期对不同基因型油菜产量特性的影响

研究了 3个不同基因型油菜品种的产量与产量构成因素的特征及其与光温条件的生态关系 ,探讨了长江中游油菜区多熟种植条件下提高油菜单产的可行途径 .结果表明 ,因播种期不同而导致油菜生育期间光温条件的不同 ,使各播期产量及产量构成因素产生了较大的差异 .油菜单株角果数、角果皮指数、主茎产量、分枝产量均与积温和日照时数呈极显著正相关 ,而千粒重和SNPA与之无关 .播种期对油菜产量的影响主要是通过影响分枝产生及其产量形成而实现的 .因此多熟制油菜的播种期可适当提早 ,10月 2 0日以后播种的油菜 ,其种植密度可增加至 3 75× 10 5～ 4 5× 10 5株·hm-2 ,以主茎弥补分枝小而少的不足 ,实现油菜单产的提高 .     

不同播期对不同基因l型油菜产量特性的影响

研究了3个不同基因型油菜品种的产量与产量构成因素的特征及其与光温条件的生态关系,探讨了长江中游油菜区多熟种植条件下提高油菜单产的可行途径.结果表明,因播种期不同而导致油菜生育期间光温条件的不同,使各播期产量及产量构成因素产生了较大的差异.油菜单株角果数、角果皮指数、主茎产量、分枝产量均与积温和日照时数呈极显著正相关,而千粒重和SNPA与之无关.播种期对油菜产量的影响主要是通过影响分枝产生及其产量形成而实现的.因此多熟制油菜的播种期可适当提早,10月20日以后播种的油菜,其种植密度可增加至3.75×105～4.5×105株·hm-2,以主茎弥补分枝小而少的不足,实现油菜单产的提高.     

Genome-wide association study for candidate genes controlling seed yield and its components in rapeseed (Brassica napus subsp. napus)

Genetic improvement of seed yield per plant (SY) is one of the major objectives in Brassica napus breeding programme. SY, being a complex quantitative trait is directly and indirectly influenced by yield-component traits such as siliqua length (SL), number of seeds per siliqua (NSS), and thousand seed weight (TSW). Therefore, concurrent improvement in SL, NSS and TSW can lead to higher SY in B. napus. This study was conducted to identify significant SNPs and putative candidate genes governing SY and its component traits (SL, NSS, TSW). All these traits were evaluated in a diverse set of 200 genotypes representing diversity from wide geographical locations. Of these, a set of 125 genotypes were chosen based on pedigree diversity and multi-location trait variation for genotyping by sequencing (GBS). Best linear unbiased predictors (BLUPs) of all the traits were used for genome-wide association study (GWAS) with 85,126 SNPs obtained from GBS. A total of 16, 18, 27 and 18 SNPs were found to be significantly associated for SL, NSS, TSW and SY respectively. Based on linkage disequilibrium decay analysis, 150 kb genomic region flanking the SNP was used for the identification of underlying candidate genes for each test trait. Important candidate genes involved in phytohormone signaling (WAT1, OSR1, ARR8, CKX1, REM7, REM9, BG1) and seed storage proteins (Cruciferin) were found to have significant influence on seed weight and yield. Genes involved in sexual reproduction and fertilization (PERK7, PERK13, PRK3, GATA15, NFD6) were found to determine the number of seeds per siliqua. Several genes found in this study namely ATS3A, CKX1, SPL2, SPL6, SPL9, WAT1 showed pleiotropic effect with yield component traits. Significant SNPs and putative candidate genes identified for SL, NSS, TSW and SY could be used in marker-assisted breeding for improvement of crop yield in B. napus. Genotypes identified with high SL, NSS, TSW and SY could serve as donors in crop improvement programs in B. napus.Supplementary InformationThe online version contains supplementary material available at 10.1007/s12298-021-01060-9.     

Synthesis of Ogura male sterile rapeseed (Brassica napus L.) with cold tolerance by protoplast fusion and effects of atrazine resistance on seed yield

Twenty-one cold-tolerant, male sterile Brassica napus somatic hybrids were produced by protoplast fusion. The fusion partners were a coldsensitive, Ogura cytoplasmic male sterile cauliflower inbred (B. oleracea var. botrytis inbred NY7642A) and a cold-tolerant, fertile canola-type B. rapa cv. Candle. Hybridity was confirmed by morphology, isozyme expression, flow cytometry, and DNA hybridization. Organellar analyses revealed a very strong bias for Brassica over Raphanus chloroplasts. Cold tolerance was confirmed by cold chamber studies and chloroplast DNA analyses. Good female fertility with 21.4 ± 3.1 seeds/pod was observed in the field using natural pollination vectors. Total seed yield was significantly greater for the atrazine-sensitive somatic hybrids produced in this study than for atrazine-resistant isolines.Abbreviations CMS cytoplasmic male sterility - IA iodoacetate - cpDNA chloroplast DNA     

Isolation of genomic DNA from defatted oil seed residue of rapeseed (Brassica napus)

A simple protocol for obtaining pure, restrictable and amplifiable megabase genomic DNA from oil-free seed residue of Brassica napus, an important oil seed plant, has been developed. Oil from the dry seeds was completely recovered in an organic solvent and quantified gravimetrically followed by processing of the residual biomass (defatted seed residue) for genomic DNA isolation. The isolated DNA can be cut by a range of restriction enzymes. The method enables simultaneous isolation and recovery of lipids and genomic DNA from the same test sample, thus allowing two independent analyses from a single sample. Multiple micro-scale oil extraction from the commercial seeds gave approximately 39% oil, which is close to the usual oil recovery from standard oil seed. Most of the amplified fragments were scored in the range of 2.5 to 0.5 kb, best suited for scoring as molecular diagnostics.     

Genetic enhancement of Brassica napus seed quality

The ultimate value of the Brassica napus (canola) seed is derived from the oil fraction, which has long been recognized for its premium dietary attributes, including its low level of saturated fatty acids, high content of monounsaturated fatty acids, and favorable omega-3 fatty acid profile. However, the protein (meal) portion of the seed has also received favorable attention for its essential amino acids, including abundance of sulfur-containing amino acids, such that B. napus protein is being contemplated for large scale use in livestock and fish feed formulations. Efforts to optimize the composition of B. napus oil and protein fractions are well documented; therefore, this article will review research concerned with optimizing secondary metabolites that affect the quality of seed oil and meal, from undesirable anti-nutritional factors to highl value beneficial products. The biological, agronomic, and economic values attributed to secondary metabolites have brought much needed attention to those in Brassica oilseeds and other crops. This review focuses on increasing levels of beneficial endogenous secondary metabolites (such as carotenoids, choline and tochopherols) and decreasing undesirable antinutritional factors (glucosinolates, sinapine and phytate). Molecular genetic approaches are given emphasis relative to classical breeding.     

Effect of enhanced UV-B radiation on pollen quantity,quality, and seed yield in Brassica rapa (Brassicaceae)

We conducted three experiments to examine the influence of ultraviolet-B radiation (UV-B; 280–320 nm) exposure on reproduction in Brassica rapa (Brassicaceae). Plants were grown in a greenhouse under three biologically effective UV-B levels that simulated either an ambient stratospheric ozone level (control), 16% (“low enhanced”), or 32% (“high enhanced”) ozone depletion levels at Morgantown, WV, USA in mid-March. In the first experiment, we examined whether UV-B level during plant growth influenced in vivo pollen production and viability, and flower production. Pollen production and viability per flower were reduced by ≈50% under both enhanced UV-B levels relative to ambient controls. While plants under high-enhanced UV-B produced over 40% more flowers than plants under the two lower UV-B treatments, whole-plant production of viable pollen was reduced under high-enhanced UV-B to 17% of that of ambient controls. Whole-plant production of viable pollen was reduced under low-enhanced UV-B to 34% of ambient controls. In the second experiment, we collected pollen from plants under the three UV-B levels and examined whether source-plant UV-B exposure influenced in vitro pollen germination and viability. Pollen from plants under both enhanced-UV-B treatments had initially lower germination and viability than pollen from the ambient level. After in vitro exposure to the high-enhanced UV-B levels for 6 h, viability of the pollen from plants grown under ambient UV-B was reduced from 65 to 18%. In contrast, viability of the pollen from plants grown under both enhanced UV-B treatments was reduced to a much lesser extent: only from ≈43 to 22%. Thus, ambient source-plant pollen was more sensitive to enhanced UV-B exposure. In the third experiment, we used pollen collected from source plants under the three UV-B levels to fertilize plants growing under ambient-UV-B levels, and assessed subsequent seed production and germination. Seed abortion rates were higher in plants pollinated with pollen from the enhanced UV-B treatments, than from ambient UV-B. Despite this, seed yield (number and mass) per plant was similar, regardless of the UV-B exposure of their pollen source. Our findings demonstrate that enhanced UV-B levels associated with springtime ozone depletion events have the capacity to substantially reduce viable pollen production, and could ultimately reduce reproductive success of B. rapa.     

QTL and epistatic analyses of heterosis for seed yield and three yield component traits using molecular markers in rapeseed (Brassica napus L.)

Aiming to explore the basis of heterosis in rapeseed, QTLs for yield and three yield component traits were mapped and the digenic interactions were detected in an F₂ population derived from a cross between two elite rapeseed lines, SI-1300 and Eagle, in this study. Twenty-eight QTLs were detected for the four yield traits, with only two of them detected simultaneously in the Wuhan and Jingmen environments. Additive, partial dominance, dominance, and overdominance effects were all identified for the investigated traits. Dominance (including partial dominance) was shown by 55% of the QTLs, which suggests that dominance is a major genetic basis of heterosis in rapeseed. At the P ?? 0.01 level with 1000 random permutations, 108 and 104 significant digenic interactions were detected in Wuhan and Jingmen, respectively, for the four yield-related traits using all possible locus pairs of molecular markers. Digenic interactions, including additive by additive, additive by dominance, and dominance by dominance, were frequent and widespread in this population. In most cases (78.3%), the interactions occurred among marker loci for which significant effects were not detected by single-locus analysis. Some QTLs (57.1%) detected by single-locus analysis were involved in epistatic interactions. It was concluded that epistasis, along with dominance (including partial dominance), is responsible for the expression of heterosis in rapeseed.     

Spontaneous gene flow from rapeseed (Brassica napus) to wild Brassica oleracea

Research on the environmental risks of gene flow from genetically modified (GM) crops to wild relatives has traditionally emphasized recipients yielding most hybrids. For GM rapeseed (Brassica napus), interest has centred on the 'frequently hybridizing' Brassica rapa over relatives such as Brassica oleracea, where spontaneous hybrids are unreported in the wild. In two sites, where rapeseed and wild B. oleracea grow together, we used flow cytometry and crop-specific microsatellite markers to identify one triploid F1 hybrid, together with nine diploid and two near triploid introgressants. Given the newly discovered capacity for spontaneous introgression into B. oleracea, we then surveyed associated flora and fauna to evaluate the capacity of both recipients to harm cohabitant species with acknowledged conservational importance. Only B. oleracea occupies rich communities containing species afforded legislative protection; these include one rare micromoth species that feeds on B. oleracea and warrants further assessment. We conclude that increased attention should now focus on B. oleracea and similar species that yield few crop-hybrids, but possess scope to affect rare or endangered associates.     

Segregation in the progeny of transformed rapeseed (Brassica napus)

The primary transformant of spring rapeseed cv. HM-81 contained TL- and TR-DNA of agropine plasmid pRi ofAgrobacterium rhizogenes 15834. The presence of TL-DNA corresponds to visible transformed phenotype in its progeny; the leaves are wrinkled and the plants are shorter than normal plants. R₁ R₂ and R₃ generations have mostly transformed phenotype. The normal phenotype appears in a low frequency in F₁ generation. Autogamised F₁ plants segregate in F₂ transformed and normal phenotype in 3:1 ratio. It is possible to suppose that TL-DNA is present in two differentloci of one pair of homologic chromosomes. The recombination frequency is 12 % (microsporogenesis) or 6 % (microsporogenesis and macrosporogenesis). In some crosses the transformed phenotype has a maternal type of inheritance. Maternal inheritance influences also several growth characteristics,e.g. length of plants and number of seeds/pods.     

Environmental variation for outcrossing rate in rapeseed (Brassica napus)

Summary Rapeseed (Brassica napus) is a predominantly selfpollinated crop with about one-third outcrossing. The outcrossing rate may be influenced by environmental factors, and hence changes in the heterozygosity level of a variety may occur during multiplication. In an investigation on environmental variation in outcrossing, we estimated the outcrossing rate in the Swedish spring rapeseed cv Topas by isozyme analysis and found that outcrossing varied from 12% to 47% over five locations in Sweden, Denmark and Germany. Among flowers at different positions on the same plant, average outcrossing varied from 11% at the top to 39% at the bottom of the plant. In conclusion, environmental factors can greatly influence the outcrossing rate in rapeseed, and an investigation therefore merit further studies.     

Association mapping for phenological, morphological, and quality traits in canola quality winter rapeseed (Brassica napus L.)

QTL mapping by association analysis has recently gained interest in plant breeding research as an alternative to QTL mapping in segregating populations from biparental crosses. In a first experiment on whole-genome association analysis in rapeseed, 684 mapped AFLP markers were tested for association with 14 traits in a set of 84 canola quality winter rapeseed cultivars. For association analysis a general linear model was used. By testing significance of marker-trait associations against a false discovery rate of 0.2, between 1 and 34 associated markers were found for 10 of the 14 traits. Taking into account linkage disequilibrium between the significant markers, these markers represent between 1 and 22 putative QTL for the respective traits. The minimum phenotypic variance explained by the QTL for the different traits ranged from 15% to 53%. A subset of 27 markers were significantly associated with two or more traits. These markers were predominantly shared between traits that were significantly correlated at the phenotypic level. The results show clearly that in rapeseed, QTL mapping by association analysis is a viable alternative to QTL mapping in segregating populations.     

Fitness of hybrids between rapeseed (Brassica napus) and wild Brassica rapa in natural habitats

Fitness of hybrids between genetically modified (GM) crops and wild relatives influences the likelihood of ecological harm. We measured fitness components in spontaneous (non-GM) rapeseed x Brassica rapa hybrids in natural populations. The F1 hybrids yielded 46.9% seed output of B. rapa, were 16.9% as effective as males on B. rapa and exhibited increased self-pollination. Assuming 100% GM rapeseed cultivation, we conservatively predict < 7000 second-generation transgenic hybrids annually in the United Kingdom (i.e. approximately 20% of F1 hybrids). Conversely, whilst reduced hybrid fitness improves feasibility of bio-containment, stage projection matrices suggests broad scope for some transgenes to offset this effect by enhancing fitness.     

Genetic control of linolenic acid concentration in seed oil of rapeseed (Brassica napus L.)

Summary Results from a diallel mating of two rapeseed lines with distinctly different linolenic acid concentration show that this trait is mainly under control of nuclear genes of the embryo. However, significant differences in reciprocal F₁, BC₁ and BC₂ indicate maternal control, which is realized by interaction between maternal genotype and nuclear genes of the embryo. Additionally, temperature exerts considerable influence on the degree of maternal control. Since no reciprocal differences are detectable in F₂, cytoplasmic factors seem not to be involved in the inheritance of linolenic acid concentration. Hypotheses on the physiological nature of maternal control of this trait are discussed.     

Effects of simulated acid rain on oilseed rape (Brassica napus) physiological characteristics at flowering stage and yield

A pot experiment was conducted to study the effects of different acidity simulated acid rain on the physiological characteristics at flowering stage and yield of oilseed rape (B. napus cv. Qinyou 9). Comparing with the control (pH 6.0), weak acidity (pH = 4.0-5.0) simulated acid rain stimulated the rape growth to some extent, but had less effects on the plant biomass, leaf chlorophyll content, photosynthetic characteristics, and yield. With the further increase of acid rain acidity, the plant biomass, leaf chlorophyll content, photosynthetic rate, antioxidative enzyme activities, and non-enzyme antioxidant contents all decreased gradually, while the leaf malonyldialdehyde (MDA) content and relative conductivity increased significantly. As the results, the pod number per plant, seed number per pod, seed weight, and actual yield decreased. However, different yield components showed different sensitivity to simulated acid rain. With the increasing acidity of simulated acid rain, the pod number per plant and the seed number per pod decreased significantly, while the seed weight was less affected.     

Inbreeding depression and dominance-suppression competition after inbreeding in rapeseed (Brassica napus)

Rapeseed plants, of the summer annual variety Topas, that had been selfed twice consecutively were compared to outcrossed half-sibs for inbreeding depression in a rapeseed population at mating equilibrium. The effect of dominance-suppression competition was included in the effect of inbreeding. Both female-and male-fitness characters showed significant inbreeding depression. Biomass decreased 17% with inbreeding and was highly correlated with seed weight. The total number of flowers decreased 15% with inbreeding. There was a significant effect of lines. The possible importance of experimental design in studies that estimate inbreeding depression is discussed.     

Extent and structure of linkage disequilibrium in canola quality winter rapeseed (Brassica napus L.)

Linkage disequilibrium was investigated in canola quality winter rapeseed to analyze (1) the prospects for whole-genome association analyses and (2) the impact of the recent breeding history of rapeseed on linkage disequilibrium. A total of 845 mapped AFLP markers with allele frequencies ≥0.1 were used for the analysis of linkage disequilibrium in a population of 85 canola quality winter rapeseed genotypes. A low overall level of linkage disequilibrium was found with a mean r ² of only 0.027 over all 356,590 possible marker pairs. At a significance threshold of P = 2.8 × 10⁻⁷, which was derived by a Bonferroni correction from a global α-level of 0.1, only 0.78% of the marker pairs were in significant linkage disequilibrium. Among physically linked marker pairs, the level of linkage disequilibrium was about five times higher with more than 10% of marker pairs in significant linkage disequilibrium. Linkage disequilibrium decayed rapidly with distance between linked markers with high levels of linkage disequilibrium extending only for about 2 cM. Owing to the rapid decay of linkage disequilibrium with distance association analyses in canola quality rapeseed will have a significantly higher resolution than QTL analyses in segregating populations by interval mapping, but much larger number of markers will be necessary to cover the whole genome. A major impact of the recent breeding history of rapeseed on linkage disequilibrium could not be observed.     

Chloroplast transformation of rapeseed (Brassica napus) by particle bombardment of cotyledons

A protocol for chloroplast transformation of an elite rapeseed cultivar (Brassica napus L.) was developed based on optimized conditions for callus induction and regeneration from cotyledonary tissues. Comparison of six different media with three elite cultivars showed that B5 medium plus 3 mg/l AgNO₃ supplemented with 0.6 mg/l 2,4-dichlorophenoxyacetic acid and 0.2 mg/l 6-furfurylaminopurine was optimal for callus formation and maintenance without differentiation, while the medium suitable for regeneration was B5 medium supplemented with 1 mg/l 6-benzylaminopurine, 1 mg/l 6-furfurylaminopurine and 0.5 mg/l α-naphthaleneacetic acid. A rapeseed-specific chloroplast transformation vector was constructed with the trnI and trnA sequences amplified from the rapeseed chloroplast genome using two primers designed according to Arabidopsis homologs. The aadA gene was used as a selection marker regulated by the ribosome-binding site from the bacteriophage T7 gene 10L, the tobacco 16S rRNA promoter and the psbA terminator. After bombardment, cotyledonary segments were cultured for callus formation on media containing 10 mg/l spectinomycin and regeneration was carried out on medium with 20 mg/l spectinomycin. Heteroplasmic plastid transformants were isolated. An overall efficiency for the chloroplast transformation was one transplastomic plant per four bombarded plates. Southern blot analyses demonstrated proper integration of the target sequence into the rapeseed chloroplast genome via homologous recombination. The expression of the aadA gene was confirmed by Northern blot analysis. Analysis of T1 transplastomic plants revealed that the transgenes integrated into the chloroplast were inheritable with a ratio of about 8%. These results suggest that rapeseed may be a suitable crop for chloroplast transformation with cotyledons as explants under appropriate conditions.     

用EST-SSR标记检测我国甘蓝型油菜品种的遗传多样性

杂交育种依然是我国油菜育种的主要方法,杂种优势的利用仍然是提高产量的重要途径.为了解我国甘蓝型油菜的遗传变异,采用16个EST-SSR标记对近年来推广的91个品种的遗传多样性进行了分析.共扩增到100个条带,其中84个多态性带,多态性比率为84%.平均每对引物扩增的带数和多态性带数分别为6.25个和5.25个.多态性信息含量(PIC)变化在0.022-0.926之间,平均为0.677,所揭示的基因型数变化于2-24之间,平均为12.44个.供试材料间遗传距离变幅较大(0.0530-0.7223之间),说明它们具有广泛的遗传变异.其中,杂交种和2000年以后育成品种的遗传基础较宽,遗传多样性分别明显高于常规品种和2000年以前育成的品种.按非加权成对平均数法(UPGMA)进行的聚类分析显示,在遗传距离为0.313处,参试材料可以分为三大类,其中,包含87份材料的第一大类在遗传距离为0.233处又可进一步分为10个亚类.聚类结果与系谱来源基本一致,比较真实反映了所用材料的遗传变异情况.