Estimation of seed weight, oil content and fatty acid composition in intact single seeds of rapeseed ( Brassica napus} L.) by near-infrared reflectance spectroscopy

The potential of near-infrared reflectance spectroscopy (NIRS) for the simultaneous analysis of seed weight, total oil content and its fatty acid composition in intact single seeds of rapeseed was studied. A calibration set of 530 single seeds was analysed by both NIRS and gas-liquid chromatography (GLC) and calibration equations for the major fatty acids were developed. External validation with a set of 75 seeds demonstrated a close relationship between NIRS and GLC data for oleic (r = 0.92) and erucic acid (r = 0.94), but not for linoleic (r = 0.75) and linolenic acid (r = 0.73). Calibration equations for seed weight and oil content were developed from a calibration set of 125 seeds. A gravimetric determination was used as reference method for oil content. External validation revealed a coefficient of correlation between NIRS and reference methods of 0.92 for both traits. The performance of the calibration equations for oleic and erucic acid was further studied by analysing two segregating F2 seed populations not represented in the calibration set. The results demonstrated that a reliable selection for both fatty acids in segregating populations can be made by using NIRS. We concluded that a reliable estimation of seed weight, oil content, oleic acid and erucic acid content in intact, single seeds of rapeseed is possible by using NIRS technique. This revised version was published online in July 2006 with corrections to the Cover Date.     

Analysis of total glucosinolate content and individual glucosinolates in Brassica spp. by near-infrared reflectance spectroscopy

This study was conducted to test the applicability of near-infrared reflectance spectroscopy (NIRS) for estimating the total glucosinolate (GSL) content in samples of intact seed from a wide range of Brassica species, and to develop calibration equations to estimate simultaneously the percentage of individual GSLs. A total of 290 samples from 15 different Brassica species were scanned by NIRS and analysed for glucosinolate content by high-pressure liquid chromatography (HPLC). A calibration equation for total GSL content was developed using 270 samples of 14 species in a range between 6 and 193μmol/g seed, resulting in an r² of 0.99 in calibration and cross-validation, and 0.95 in independent validation with 20 samples of Brassica rapa, a species not represented in the calibration. Furthermore, calibration equations to estimate the relative amount (mol/mol) of progoitrin, sinigrin, and gluconapin were successfully developed (r² > 0.85 in cross-validation) and validated with samples from species not included in the calibration. It was also possible to discriminate between entries with high and low values of glucoiberin, 4-hydroxyglucobrassicin and glucoerucin.     

Genetic variation for sinapate ester content in winter rapeseed (Brassica napus L.) and development of NIRS calibration equations

Increasing the meal and protein quality of winter rapeseed (Brassica napus L.) for food and feed purposes is gaining importance in rapeseed breeding programmes. Rapeseed meal has a high content of phenolic acid esters, mainly sinapate esters, which have been shown to cause a dark colour and a bitter taste in rapeseed meal and derived protein products. The aim of the present study was to analyse the genetic variation for individual and total sinapate ester content, to develop Near Infrared Reflectance Spectroscopic (NIRS) calibrations, and to identify genotypes with a low sinapate ester content after testing in the field. The following sinapate esters were analysed by HPLC: sinapoylcholine (sinapine), sinapoylglucose, and a minor group of ‘other sinapate esters’ which includes free sinapate. A genotypically diverse set of seed samples of winter oilseed rape (old and new cultivars, breeding lines, resynthesized rapeseed) from different years and locations was collected, their NIRS spectra recorded and the samples were further analysed by HPLC. The complete NIRS calibration seed sample set (n = 575) showed a large variation in total sinapate ester content, ranging from 3.2 to 12.7 mg sinapate equivalents per g seeds. The NIRS calibration equations showed high fractions of explained variances in cross validation () ranging from 0.75 (other sinapate esters) to 0.85 (sinapoylglucose). The standard errors of cross validation (SECV) ranged from 0.38 (other sinapate esters) to 0.70 mg/g seed (total sinapate esters). In validation and in independent validations the predicted results were not always acceptable, indicating that the NIRS calibrations need to be extended by analysing samples from new populations. Following replicated field experiments, a doubled haploid line obtained from the old Dutch cultivar Mansholts’ Hamburger Raps, and related DH lines from the cross DH Mansholts’ × Express were confirmed to have a 30–40% lower sinapate ester content compared to check cultivars.     

甘蓝型油菜种子中几种主要脂肪酸含量的遗传

通过对甘蓝型油菜杂交组合(华油8号×Altex)正反交 F_1、F_2、回交一代以及相应的杂交亲本种子中几种主要脂肪酸含量的气相色谱测定,研究了芥酸、廿碳烯酸、油酸、亚油酸和亚麻酸含量的遗传以及这几种脂肪酸之间的相互关系。研究结果表明:种子中的芥酸、廿碳烯酸和油酸含量都是由胚基因型决定的,这三种脂肪酸含量受到一个共同     

Inheritance of long chain fatty acid content in rapeseed (Brassica napus L.)

A large industrial oil market has recently developed for high erucic acid (>500 g kg⁻¹) rape (Brassica napus L.) cultivars. This research was conducted to: (i) determine genetic effects for five fatty acids, (ii) determine if maternal effects influence fatty acid content of progeny, and (iii) estimate correlations among fatty acid contents in hybrid progeny. Lines with very high erucic acid content and very low erucic acid content were used to develop eight generations to estimate additive, dominance, and epistatic effects for fatty acid content using Generation Means Analyses. Mean oleic, linoleic, linolenic, eicosenoic and erucic acid content differed among generations and additive genetic effects were important for control of all five fatty acids, contributing from 84% to 97% of the total sums of squares for each fatty acid. Epistasis was observed in the inheritance of eicosenoic acid. Maternal effects were not detected.     

Identification of genetic variation in Brassica napus seeds for tocopherol content and composition using near‐infrared spectroscopy technique

Tocopherol is an essential fat‐soluble nutrient for humans. Increasing the tocopherol content in Brassica napus seeds can add value to rapeseed vegetable oil; this has become an important breeding target. However, there is no efficient and non‐destructive method for selecting rapeseed accessions with high tocopherol contents. Here, we report the first near‐infrared reflectance spectroscopy (NIRS)‐based technique for predicting rapeseed tocopherol content. Individual seed tocopherol compositions were estimated from 373 rapeseed genotypes of different origins. This method and chemical methods produced comparable predicted values of the tocopherol constituents in the seeds. Three equations were generated for the prediction of tocopherol content by using a modified partial least squares (MPLS) model. The total tocopherol content for the determination coefficient of cross‐validation (R²_cv) (0.74), determination coefficient (RSQ) (0.76) and one minus the ratio of unexplained variance to total variance (1‐VR) (0.65) values indicates a strong correlation between the calibration and validation sets. Overall, our model confirmed the NIRS method as feasible for predicting tocopherol content in rapeseed and as an efficient screening tool for future breeding programs.     

Genetic control of fatty acid composition in seed oil of Sinapis alba L.

Summary Inheritance of fatty acid composition was studied in an F1 diallel cross in Sinapis alba. Crosses were made among accessions having contrasting amounts of oleic (C18:1) and erucic (C22:1) acid. Concentrations of oleic, linoleic (C18:2), eicosenoic (C20:1) and erucic (C22:1) acids were determined by gas-chromatography for each mating combination. Genetic analysis confirmed that the composition of the fatty acids was controlled mainly by the nuclear genes of the embryo. Additive gene action with partial dominance for the reducing alleles was noted for oleic and linoleic acids, while erucic acid showed an additive mode of inheritance with partial dominance for the enhancing alleles. Positive heterosis was demonstrated for eicosenoic acid content. Erucic acid content was strongly negatively correlated with oleic acid, suggesting a genetic interdependence between the two fatty acids. Broad-sense and narrow-sense heritability estimates for each of oleic, linoleic and erucic acids were very high, due to low between-plants non-genetic component of variance.Contribution No. 3662-E, 1992 series.     

Evaluation of Near Infra-red Reflectance Spectroscopy for Predicting Grain and Stover Quality Traits in Maize

The prediction of grain and stover quality parameters in maize {Zea mays L.) by near infra-red reflectance spectroscopy (NIRS) was studied. A total of 110 grain and 135 stover samples originating from different genotypes and environments were assayed. Calibration equations for content of crude protein (CP), crude fat (CF), starch (ST), and water soluble carbohydrates (WSC) in grain were obtained by multiple linear regression of known manual values on NIRS data from the odd numbered samples. Calibrations for CP, acid detergent fibre (ADF), in vitro digestible organic matter according to the Tilley & Terry (IVDOM-T & T) and the gas production (IVDOM-Gp) method, respectively, and metabolizable energy (ME) in stover were developed analogously. Equations were validated with the evennumbered .samples and for ME additionally with the 1584 stover samples from an experiment with 66 F₁ hybrids tested in six environments. The coefficients of multiple determination (R²) of the prediction equations ranged from 0.80 for IVDOM-Gp and ME in stover to 0.94 for CP in grain. Standard errors of calibration (SEC) and prediction (SEP) were in most cases not higher than commonly reported for conventional manual assays. With regard to the correct ranking of hybrids, prediction equations for ME applied well to stover samples from other environments with one exception. We concluded that NIRS can evaluate the quality traits investigated to a similar degree to that of conventional methods of analysis. Since NIRS is simple and safe to operate and allows rapid screening of several quality traits simultaneously, it should be particularly attractive for breeding purposes.     

工业专用型高芥酸油菜新品种选育

高芥酸油菜品种是在工业上具有广泛用途的专用型品种。本研究通过两个常规芥酸品种杂交,采用以单株和单粒筛选相结合对芥酸含量正向选择为核心的技术,育成了芥酸含量达60%的甘蓝型高芥酸油菜新品种高芥1号。同时阐明了在高芥酸含量背景下各种脂肪酸间的相关性,为相关育种提供了—些理论依据。     

应用近红外反射光谱法整粒测定小样品油菜籽含油量的研究

以完整油菜籽为样品,通过光谱预处理和回归统计方法来优化油菜籽含油量的近红外反射光谱法所建立回归方程,同时采用三种不同用量的样品杯进行近红外扫描分析.结果表明,光谱预处理对较正结果影响较大,不同光谱数学处理以3级导数处理较好,综合考虑定标组和检验组的效果,采用"2,3,3,1"、 "2,6,6,1"和"3,3,3,1"的三种处理组合较好     

The measurements of acid detergent fibre in rapeseed by visible and near-infrared spectroscopy

Visible and near‐infrared spectroscopy (NIRS) calibrations for acid detergent fibre (ADF) in intact rapeseed (Brassica napus L.) were performed for two different sample volumes (10 ml, 500 seeds approximately; and 1 ml, 50 seeds approximately). The inclusion of brown and yellow‐seeded cultivars in this work has allowed the whole range of ADF currently described in the literature for this character to be covered. Chemometric techniques have been used for developing calibration equations for both procedures when measuring the two different seed sample volumes. On the basis of the coefficient of determination in the cross‐validation (R²cv) obtained for the 10 and 1 ml assays (0.80 and 0.73), and SECV/SEL ratios (2.30 and 2.57), respectively, both equations showed an accuracy sufficient for screening purposes in an ADF range from 6.80 to 13.46% dry wt, which is presented in this work.     

甘蓝型油菜主要脂肪酸组成的QTL定位

应用RAPD、SSR和SRAP技术, 对甘蓝型油菜低芥酸品系APL01与高芥酸品系M083杂交组合的BC1F1群体进行检测, 获得251个分子标记, 构建了19个连锁群组成的分子标记遗传图谱; 应用WinQTLCart 2.0对油菜主要脂肪酸组成进行QTL扫描, 获得与棕榈酸含量相关的QTL 5个, 分别位于N3、N8、N10和N13连锁群, 其中效应值较大的主效QTL qPA8-1和qPA13分别可解释棕榈酸含量表型变异的11.31%和14.47%。获得与硬脂酸含量相关的QTL 3个, 分别位于N1、N8和N16连锁群, 其中效应值较大的主效QTL qST16可解释硬脂酸含量表型变异的12.22%。获得与油酸含量相关的QTL 2个, 位于N8和N13连锁群, 均为主效QTL, 其中qOL8位于N8连锁群的m11e37b~A0226Ba267区间, 可解释油酸含量表型变异的11.73%, qOL13位于N13连锁群的m18e46~m20e25a区间, 可解释表型变异的27.14%。获得与亚油酸含量相关的QTL 3个, 其中主效QTL qLI8-1位于N8连锁群, 可解释亚油酸含量表型变异的13.25%。获得与亚麻酸含量相关的QTL 3个, 效应值均较小, 属微效QTL。获得与廿碳烯酸含量相关的QTL 4个, 分别位于N8、N13和N15连锁群, 其中主效QTL qEI8-1、qEI8-2和qEI13分别可解释廿碳烯酸含量表型变异的12.20%、10.22%和11.14%。获得与芥酸含量相关的QTL 2个, 位于N8和N13连锁群, 均为主效QTL, 其中qER8位于N8连锁群的m11e37b~A0226Ba267区间, 可解释芥酸含量表型变异的16.74%; qER13位于N13连锁群的A0301Bb398~m18e46区间, 可解释芥酸含量表型变异的31.32%。在N8连锁群的分子标记m11e27b附近及N13连锁群的分子标记m18e46附近存在多个主要脂肪酸的主效QTL, 这些标记可用于油菜脂肪酸改良的分子标记辅助选择。     

Near infrared reflectance spectroscopy analysis of digestibility traits of maize stover

Summary Near infrared reflectance spectroscopy (NIRS) is emerging as a potentially useful tool in breeding plants for quality traits. Information is lacking, however, on its use in forage maize (Zea mays L.). The objectives of the present investigation were to evaluate the prediction of digestibility traits of maize stover using NIRS technique and to study the effect of laboratory (Lab) and NIRS assays on the estimates of variation and covariation. Twelve inbred lines, 66 diallel crosses among them and eight hybrid checks were evaluated at silage and grain harvests for 2 years at two agro-climatically diverse locations in the Federal Republic of Germany. Standard methods were used for Lab analysis of in vitro digestible organic matter (IVDOM), neutral detergent fibre (NDF), acid detergent fibre (ADF) and acid detergent lignin (ADL). In NIRS evaluation, calibration equations were developed by modified stepwise regression. The standard error of calibration was 2.5, 1.7, 1.4 and 0.4 for IVDOM, NDF, ADF and ADL, respectively. The coefficient of multiple determination was high (0.9) except for ADL. The validation statistics (standard error and correlation coefficient) were similar. In the diallel crosses, the estimates of variation (heritabilities in broad and narrow sense, genotypic and error coefficients of variation), generally, did not vary appreciably and consistently in the comparisons between Lab and NIRS methods particulary at silage harvest. Simple and rank correlations between Lab and NIRS analyses were positive and significant. These correlation coefficients based on the mean performance of the diallel crosses at silage harvest were >0.9 and at least 16 hybrids were common between the two analyses, among the upper one-third or lower one-third (22) hybrids. The study showed that NIRS analysis should be useful in maize breeding programmes wherein a large number of genotypes need to be evaluated.Abbreviations NIRS near infrared reflectance spectroscopy - Lab laboratory - IVDOM in vitro digestible organic matter - NDF neutral detergent fibre - ADF acid detergent fibre - ADL acid detergent lignin - SEC standard error of calibration - SEV standard error of validation - SD standard deviation - r simple correlation coefficient - r_s rank correlation coefficient - R² coefficient of multiple determination - h_b ² heritability in broad sense - h_n ² heritability in narrow sense - CV_g genotypic coefficient of variation - CV_e error coefficient of variation - GCA general combining ability effect     

Genetic variation in phytosterol content of winter rapeseed (Brassica napus L.) and development of NIRS calibration equations

Rapeseed oil is one of the richest natural sources of phytosterols, known to reduce the LDL-cholesterol levels, one of the major cardiovascular disease risk factors. Increasing the phytosterol content in rapeseed could give an added value to the oil and derived products. Our objective was to analyse the genetic variation of phytosterol content in modern winter rapeseed cultivars and resynthesized lines following field experiments and to develop a near-infrared reflectance spectroscopic (NIRS) calibration for high throughput estimation of phytosterol content. Phytosterol content as analysed by gas–liquid chromatography ranged from 3565 to 4800 mg/kg seed for modern cultivars and from 2079 to 4329 mg/kg seed for resynthesized lines. The NIRS calibration showed a high fraction of explained variance in cross-validation of 0.81 for total phytosterol content and the standard error of cross-validation was 241 mg/kg. The results show that the cultivars contain already high phytosterol contents. The NIRS calibrations developed for total phytosterol content should be useful for germplasm screening and in breeding programmes aimed at increasing the phytosterol content in rapeseed.     

Evaluation of microspore-derived embryos as models for studying lipid biosynthesis in seed of rapessed (Brassica napus L.)

Summary Microspore culture of rapeseed (Brassica napus L.) has provided a powerful tool not only for breeding but also in developmental studies. In this study, microspore-derived embryos (MDE) of B. napus were evaluated as a model in seed for studying accumulations of triacylglyceride (TAG) fatty acids in both a low and high erucic acid rapeseed line; and accumulations of TAG and free fatty acids (FFA) in a high erucic acid rapessed line. The accumulation patterns confirmed that MDE had a similar TAG fatty acid profile to seed during the embryo development within each genotype. The oil accumulation in MDE after 36 days in culture (DIC) approached levels similar to those in zygotic seed 25 days after flowering (DAF). Significant differences were detected in contents of both total free fatty acids and specific free fatty acids between MDE and seed. During the developmental period, total free fatty acids changed from 16% to 2.1% in MDE, but from 10.5% to 0.1% in seed. MDE had much higher percentage of free linolenic and erucic acids than seed, particularly during the late developmental stages. The current study indicated that MDE can be used as a model to study TAG and TAG fatty acids in seed but caution must be taken to study free fatty acid metabolism.     

Increasing erucic acid content in Ethiopian mustard through mutation breeding

Ethylmethane sulphonate (EMS) was applied to seeds of the Ethiopian mustard (Brassica carinata A. Braun) line C-101. Bulk samples of M₃ seeds from 8331 M₂ plants were evaluated for the fatty acid composition of their oil by near-infrared reflectance spectroscopy (NIRS) and by further gas chromatography on selected samples. A putative mutant, N2-6230, showing very low oleic acid content (4.7% vs. average of 8.6% in C-101) and erucic acid content within the range of variation of the line C-101 (40-49.3%) was identified. The M₃ progeny of this mutant showed a wide segregation for erucic acid content (39.1-57.9% vs. 41.8-50.3% in C-101), and maintained levels of oleic acid lower than in line C-101. Selection for high erucic acid content in the M₃ and M₄ generations led to the fixation of this mutation in the M₅ generation (52.2-59.3% vs. 39.0-47.6% in C-101). This is the first high erucic acid line obtained in Brassica species through mutation breeding. Its utility in future programmes to develop very high erucic acid lines is discussed.     

Nondestructive assessment of protein content in single seeds of rapeseed (Brassica napus L.) by near-infrared reflectance spectroscopy

The potential of near-infrared reflectance spectroscopy (NIRS) to detectwithin-plant differences for seed protein content was investigated. Fourhundred and fifty-one single seeds were scanned by NIRS using a specialadapter. After non-destructive NIRS scanning, the seeds were analysed forprotein content by the Dumas combustion method and a calibrationequation was developed. A validation set of 117 additional seeds fromthree individual plants from the cultivars Bristol, Lirajet and Maplus wasanalysed for protein content both by NIRS and combustion. The coefficientof determination between NIRS and combustion values in the validation setwas 0.94, with a standard error of performance (SEP) of 0.77% and aratio of the SEP to the standard deviation (SD) of the validation set of0.28. The coefficient of variation (CV) for seed protein content inindividual plants, as determined by the combustion method, was 11.7%for Bristol, 8.9% for Lirajet, and 9.5% for Maplus. The comparison ofsuch variation with the standard error (SE) of NIRS analysis, defined as thecombination of the SE of the combustion method and the SEP of NIRScalibration equation, revealed that the maximum explainable variance withinindividual plants that can be detected using NIRS analysis of proteincontent in single seeds was 0.86 for Bristol, 0.83 for Lirajet, and 0.85 forMaplus. These results demonstrated that NIRS is a powerful tool fornon-destructive assessment of within-plant variation for seed protein contentin rapeseed.     

Genetics of the Erucic Acid Content in Interspecific Hybrids of Ethiopian Mustard (Brassies carinata Braun) and Rapeseed (B. napus L.)

Ethiopian mustard (Brassica carinata Braun) is a potential oil crop in which genes for low erucic acid content of the seed oil have not yet been found. In order to solve this problem the potential of rapeseed (B. napus L.) varieties as a source of these genes has been tested. Reciprocal F₁ hybrids between B. carinata and a low erucic acid variety of B. napus, F₂, and backcrosses with B. carinata were obtained. The fatty acid composition was determined in half seeds of F₁ and segregating generations from reciprocal interspecific crosses. The genetic analysis indicated that the erucic acid content of the seed oil of B. carinata is controlled by two genes with no dominance and additive in action.     

Fatty Acid Composition of Oil from Erucic Acid-Free Summer Rape Seed (Brassica napus L.) in Relation to Nitrogen Nutrition and Raceme Position

A greenhouse study was conducted to determine the effect of nitrogen supply (30, 100 or 170 ppm N) and raceme position on the fatty acid composition of oil extracted from erucic acid-free summer rape seed ( Brassica napus cv. Callypso ). The seven fatty acids analyzed for include palmitic, palmitolcic, stearic, oleic, linoleic, linolemc, and eicosenoic acids; of which oleic (59.54–64.84 %) and palmitoleic (0.36–0.4 %) acids were the highest and lowest levels respectively. Generally, N nutrition influenced fatty acid pattern only to a little extent. Palmitic, palmitoleic and stearic acid levels were increased by 170 ppm N, depending on raceme position, but oleic and linolenic acids were unaffected. Similarly, 170 ppm N produced the highest fatty acid levels in seeds on the lower portions of racemes, with the exception of oleic acid. This was also true in the case of the upper portions of racemes, except that 30 ppm N produced the highest levels of oleic and linoleic acids in rape seeds. Under the optimum N supply level (i.e. 100 ppm N), position of raceme on the rape plant did not greatly influence the levels of different fatty acids in lipids.     

Combination of resistance to Verticillium longisporum from zero erucic acid Brassica oleracea and oilseed Brassica rapa genotypes in resynthesized rapeseed (Brassica napus) lines

Resynthesized (RS) forms of rapeseed (Brassica napus L.; genome AACC, 2n = 38) generated from interspecific hybridization between suitable genotypes of its diploid progenitors Brassica rapa L. (syn. campestris; genome AA, 2n = 20) and Brassica oleracea L. (CC, 2n = 18) represent a potentially useful resource to introduce resistance against the fungal pathogen Verticillium longisporum into the gene pool of oilseed rape. Numerous cabbage (B. oleracea) accessions are known with resistance to V. longisporum; however, B. oleracea generally has high levels of erucic acid and glucosinolates in the seed, which reduces the suitability of resulting RS rapeseed lines for oilseed rape breeding. In this study resistance against V. longisporum was identified in the cabbage accession Kashirka 202 (B. oleracea convar. capitata), a zero erucic acid mutant, and RS rapeseed lines were generated by crossing the resistant genotype with two spring turnip rape accessions (B. rapa ssp. olerifera) with zero erucic acid. One of the resulting zero erucic acid RS rapeseed lines was found to have a high level of resistance to V. longisporum compared with both parental accessions and with B. napus controls. A number of other zero erucic acid RS lines showed resistance levels comparable to the parental accessions. In the most resistant RS lines the resistance and zero erucic acid traits were combined with variable seed glucosinolate contents. Erucic acid‐free RS rapeseed with moderate seed glucosinolate content represents an ideal basic material for introgression of quantitative V. longisporum resistance derived from B. oleracea and B. rapa into elite oilseed rape breeding lines.