不同甘蓝型油菜高含油量种质资源的脂肪酸成分分析

甘蓝型油菜是我国最为重要的油料作物之一.目前我国面临着植物油严重不足的局面.提高含油量是目前甘蓝型油菜育种的主要方向之一.目前.我国育种工作者已经筛选出了大量含油量超过50%的甘蓝型油菜种质.本文对两种类型的高含油量甘蓝型油菜种质进行了脂肪酸组分分析,结果显示芥酸含量高的种质和芥酸含量低的种质的脂肪酸在菜籽油中变化范围和平均值明显不同.油酸和亚油酸在低芥酸高含油量甘蓝型油菜种质中的含量较高,约占整个油分的80%左右,其他脂肪酸组分则在10%以下;而芥酸则是高芥酸高含油量种质的最主要的成分,约占整个油分的44%左右,油酸、亚油酸等的含量则均在10%左右.相关性分析表明:芥酸和其他脂肪酸之间呈现显著负相关,因此.可以通过降低高芥酸油菜种质中的芥酸含量来提高比如油酸和亚油酸的含量.从而使这些种质的油分更适合人类的健康.     

羽衣甘蓝种子主要品质性状的表现及其相关性

随机挑选148份羽衣甘蓝种质资源和高世代材料,分析了成熟种子的含油量、蛋白质、硫苷和7种主要脂肪酸成分的表现特征及其相关性。结果表明:羽衣甘蓝成熟种子平均含油量为29.48%,平均蛋白质含量为45.13%,含油量和蛋白质总量为74.61%。硫苷含量的变幅最大,变异系数为31.72%。7种主要脂肪酸成分中,油酸和芥酸的含量较高,其次为亚油酸,棕榈酸和硬脂酸的含量较低。除硫苷含量和硬脂酸含量外,其余9个性状的表现均呈单峰正态分布。相关性分析表明,大多数性状间都具有显著或极显著的相关性,这与对甘蓝型、白菜型和芥菜型3种类型油菜的研究结果相一致。在羽衣甘蓝中存在一些优异的种质资源,通过筛选可以在油菜优质育种中加以利用。     

国际半干旱热带地区作物研究所花生微核心种质含油量及脂肪酸分析与鉴定

以国际半干旱热带地区作物研究所(ICRISAT)花生微核心种质为材料,系统分析测试含油量和脂肪酸组成。分析结果表明,ICRISAT花生微核心种质的含油量平均为51.67%,变异范围49.16%~55.44%,珍珠豆型资源的含油量高于其他类型,发掘出高油种质1份。在主要脂肪酸中,棕榈酸平均含量10.74%,变异范围7.9%~13.5%;硬脂酸2.85%,变异范围1.8%~3.9%;油酸46.36%,变异范围37.0%~64.7%;亚油酸32.86%,变异范围18.0%~40.4%;饱和脂肪酸含量19.21%,变异范围15.2%~22.1%。普通型花生的油酸含量高于其他类型,而亚油酸和棕榈酸含量低于其他类型。发掘出高油酸种质4份,低棕榈酸种质19份,低饱和脂肪酸种质7份。通过脂肪酸组成的分析,高油酸种质和低饱和脂肪酸种质均同时具备低棕榈酸的优良特性。SSR分析结果表明,这些种质的遗传差异相对较大。根据5对SSR引物的扩增结果,绘制了20份资源的分子指纹图谱,为这些优质资源的保护和有效利用奠定了基础。     

甘蓝型油菜油酸配合力的双标图分析

用GGE双标图法对甘蓝型油菜8×8完全双列杂交试验的脂肪酸组分进行分析,以期得到配合力高的亲本,为高油酸材料的转育和配组奠定基础。结果显示:(1)一般配合力(GCA)较高的亲本是父本Y511、Y539和母本Y511、Y520、Y539,而特殊配合力(SCA)较高的亲本是父本L308、Y511、Y539和母本L121、L307、L331、Y539。从GCA和SCA综合来看,Y511、Y520和Y539是配合力较高的父本,L307、L331、Y539是配合力较高的母本,父本L121和母本L308的配合力相对最低;(2)Y539是母本L121、L307、L308、L332、Y511、Y520、Y539的最佳组配父本,Y511是L331的最佳组配父本;Y520是父本L121、L307、L331、L332、Y511的最佳组配母本,Y539是L308、Y520、Y539的最佳组配母本;(3)SCA较高的组合有L307×L308、L331×L332、L332×Y511、Y520×Y511、Y539×L121等;(4)在芥酸含量为0、二十碳烯酸含量趋近于0(0.87%左右)、硬脂酸含量变幅不大(1.36%~1.75%)的遗传背景下,父本脂肪酸组分中油酸与棕榈酸、硬脂酸、亚油酸和亚麻酸均呈负相关;母本中油酸与硬脂酸呈正相关,与棕榈酸、亚油酸、亚麻酸呈负相关;(5)各杂交种脂肪酸组分的表型关系中,与油酸含量呈正相关的是硬脂酸,呈负相关的是棕榈酸,而与亚油酸、亚麻酸呈极显著负相关,这与母本脂肪酸组分之间的关系基本一致。     

油菜品质育种现状及展望

国际油菜品质改良始于20世纪60年代,以降低油菜籽中芥酸和硫代葡萄糖苷为主要目标.随着历史的不断发展,油菜的品质改良已不再局限于这两个指标.在食用油方面,已将提高油酸、亚油酸含量,降低饱和脂肪酸和亚麻酸含量作为今后的主攻方向,使之成为最健康的食用油;在工业用油方面,高芥酸和中等长度的脂肪酸改良已逐渐展开.今后常规育种、杂种优势利用和生物技术的有机结合,将使油菜品种的改良进入到一个新的阶段.     

贵州地方芝麻种质资源品质性状的分析与评价

为探究贵州芝麻种质资源的品质特征,并对地方芝麻资源进行初步鉴定与评价,本研究对73份贵州芝麻种质资源的8个品质性状进行测试分析。结果表明:(1)贵州芝麻种质资源含油量介于41.45%~52.12%之间,平均含量为49.69%。在脂肪酸组成中,油酸、亚油酸等不饱和脂肪酸的平均含量分别为35.65%和50.66%;而棕榈酸、硬脂酸等饱和脂肪酸的平均含量仅为8.40%和4.79%。此外,贵州芝麻资源中芝麻素、芝麻林素和木质素的平均含量分别为5.03 mg/g、2.63 mg/g和4.79 mg/g。8个品质性状的变异系数介于3.69%~32.62%范围内,其中芝麻素含量变异系数最大,含油量变异系数最小。而芝麻素含量、芝麻林素含量及硬脂酸含量的变异系数均大于10%,表明这3个性状在芝麻样本间存在较大差异。(2)相关性分析结果显示:含油量与油酸、芝麻素含量呈极显著正相关,与亚油酸含量呈极显著负相关;油酸含量与芝麻素含量呈极显著正相关,与亚油酸含量呈极显负相关;亚油酸含量与芝麻素含量呈极显著负相关。表明品质性状间相关性大、关联程度较高,性状间相互影响较大。(3)主成分分析将8个品质性状综合为3个主成分,分别为油酸因子、含油量因子和芝麻素因子,3个主成分因子包含了贵州芝麻种质资源品质性状的绝大部分信息,累计贡献率达96%以上。(4)在欧氏距离D=9.75处将73份贵州芝麻资源划分为6个类群:第Ⅰ类群包含2份资源、第Ⅱ类群有7份、第Ⅲ类群有12份、第Ⅳ类群有5份、第Ⅴ类群有16份、第Ⅵ类群有31份。其中第Ⅵ类群油酸含量最高,且含油量、芝麻素含量较高。本研究探明了贵州芝麻品质的特征特性,可为芝麻种质资源的利用和创新提供依据,为芝麻品种选育和遗传改良提供参考。     

油菜品质育种现状及展望

国际油菜品质改良始于20世纪60年代,以降低油菜籽中芥酸和硫代葡萄耱苷为主要目标。随着历史的不断发展,油菜的品质改良已不再局限于这两个指标。在食用油方面,已将提高油酸、亚油酸含量,降低饱和脂肪酸和亚麻酸含量作为今后的主攻方向,使之成为最健康的食用油;在工业用油方面,高芥酸和中等长度的脂肪酸改良已逐渐展开。今后常规育种、杂种优势利用和生物技术的有机结合,将使油菜品种的改良进入到一个新的阶段。     

世界红花种质的籽油脂肪酸组分评价

对引自 48个国家和地区在北京栽培的 2 0 48份红花 (CarthamustinctoriusL .)种质资源的籽油脂肪酸分析表明 ,棕榈酸、硬脂酸、油酸和亚油酸的平均含量分别为 7.30 %、1.2 8%、15 .76 %和 75 .33% ,其含量范围分别为 0 .99%～ 2 9.0 3%、0 .0 1%～ 5 .71%、5 .0 0 %～ 81.84%和 11.13%～ 88.30 %。来自不同地区的红花种质 ,各种脂肪酸的含量有较大的差异。来源于孟加拉国的红花 ,亚油酸平均含量为 5 0 .6 8% ,来源于奥地利的红花 ,亚油酸平均含量高达79.0 4%。通过评价 ,分别筛选出 10个高亚油酸和 10个高油酸的品种 ,高油酸的品种中有 3个来自孟加拉国 ,而高亚油酸的品种大多来自中国     

利用高代回交导入群体定位大豆品质性状QTL

通过对大豆品质性状的QTL定位,可以为大豆分子育种提供理论依据。本研究以中黄13(轮回亲本)×东山69(供体亲本)的142个家系的高代回交导入系群体BC2F7,利用具有多态性的113对SSR引物构建了一张涵盖除B2之外的19个大豆连锁群、总长度为1630.95 cM、平均遗传距离为14.43 cM的遗传连锁图谱。利用近红外光谱分析法(NIR)、气相色谱技术(GC)分析群体蛋白质、脂肪及5种脂肪酸主要组分含量;采用ICIM-ADD作图法定位QTL。两年共定位到34个QTL,其中与蛋白质、脂肪、硬脂酸、棕榈酸、亚油酸、油酸和亚麻酸相关的QTL分别为5、2、11、6、3、4、3个,Sat_003～Sat_147两年均被检测出与油酸含量呈负相关,5个区间与2个性状含量相关,Satt523～Sat_405与棕榈酸、蛋白质相关,Staga001～Satt658、Satt263～Satt651与油酸、硬脂酸相关,Satt228～Satt525与亚油酸、蛋白质相关,Satt658～Satt433与亚麻酸、硬脂酸相关;总体而言,定位结果在一定程度上阐明了品质性状的相关性,对大豆分子育种中优良品种的选育具有参考意义。     

不同种源山桐子果实脂肪酸组成变异分析

以采自11个种源的山桐子为材料,测定其果实脂肪酸的组成及其变异情况,结果表明：山桐子果实中不饱和脂肪酸含量较高,尤以亚油酸含量最高,11个种源的平均值为63．58％,且种源间差异显著,分宜、宜昌种源亚油酸相对含量明显高于其他9个种源;饱和脂肪酸以棕榈酸为主,11个种源山桐子果实棕榈酸差异显著,且以平武种源最高;其余脂肪酸含量均较低,变异幅度较大;种子中棕榈酸和棕榈烯酸含量明显高于果肉,而亚油酸、亚麻酸及硬脂酸含量明显低于果肉,油酸含量相近：果实不饱和脂肪酸含量依次为果肉〉全果〉种子,且变异系数及相对极差均较小,尤以果肉中最小;除硬脂酸外,山桐子果实中其他4种主要脂肪酸组分受海拔等地理环境的影响均较小。     

Eliminating expression of erucic acid-encoding loci allows the identification of “hidden” QTL contributing to oil quality fractions and oil content in <Emphasis Type="Italic">Brassica juncea</Emphasis> (Indian mustard)

Oil content and oil quality fractions (viz., oleic, linoleic and linolenic acid) are strongly influenced by the erucic acid pathway in oilseed Brassicas. Low levels of erucic acid in seed oil increases oleic acid content to nutritionally desirable levels, but also increases the linoleic and linolenic acid fractions and reduces oil content in Indian mustard (Brassica juncea). Analysis of phenotypic variability for oil quality fractions among a high-erucic Indian variety (Varuna), a low-erucic east-European variety (Heera) and a zero-erucic Indian variety (ZE-Varuna) developed by backcross breeding in this study indicated that lower levels of linoleic and linolenic acid in Varuna are due to substrate limitation caused by an active erucic acid pathway and not due to weaker alleles or enzyme limitation. To identify compensatory loci that could be used to increase oil content and maintain desirable levels of oil quality fractions under zero-erucic conditions, we performed Quantitative Trait Loci (QTL) mapping for the above traits on two independent F1 doubled haploid (F1DH) mapping populations developed from a cross between Varuna and Heera. One of the populations comprised plants segregating for erucic acid content (SE) and was used earlier for construction of a linkage map and QTL mapping of several yield-influencing traits in B. juncea. The second population consisted of zero-erucic acid individuals (ZE) for which, an Amplified Fragment Length Polymorphism (AFLP)-based framework linkage map was constructed in the present study. By QTL mapping for oil quality fractions and oil content in the ZE population, we detected novel loci contributing to the above traits. These loci did not co-localize with mapped locations of the fatty acid desaturase 2 (FAD2), fatty acid desaturase 3 (FAD3) or fatty acid elongase (FAE) genes unlike those of the SE population wherein major QTL were found to coincide with mapped locations of the FAE genes. Some of the new loci identified in the ZE population could be detected as ‘weak’ contributors (with LOD < 2.5) in the SE population in which their contribution to the traits was “masked” due to pleiotropic effects of erucic acid genes. The novel loci identified in this study could now be used to improve oil quality parameters and oil content in B. juncea under zero-erucic conditions.     

Hairpin-RNA mediated silencing of endogenous FAD2 gene combined with heterologous expression of Crambe abyssinica FAE gene causes an increase in the level of erucic acid in transgenic Brassica carinata seeds

The 3′-UTR of the FAD2 gene from Brassica carinata was cloned by PCR and used to prepare an intron-spliced hairpin RNA (ihpRNA) construct. Compared to that of the wild type (WT) background, this construct, when expressed in B. carinata, resulted in a high degree of FAD2 gene silencing accompanied by strong increases of up to 16 and 10% in oleic acid and erucic acid proportions, respectively. The increase in 18:1 was accompanied by a concomitant proportional reduction in 18:2. A second construct containing ihpRNA targeted to the endogenous FAD2 gene in addition to the heterologous Crambe abyssinica FAE gene under the control of seed specific napin promoter, was used to transform B. carinata. This approach resulted in an even greater increase in erucic acid proportions, by up to 16% in T₁ segregating seeds as compared to that of the WT control. To our knowledge, this is currently the highest accumulation of erucic acid achieved in B. carinata seeds using transgenic approaches, making it an increasingly-attractive alternative to high erucic B. napus cultivars as an industrial oil crop. Database: The nucleotide sequence reported in this paper has been submitted to the EMBL/GenBank under accession number DQ250814.     

Mapping of QTLs for oil content and fatty acid composition in Indian mustard [Brassica juncea (L.) Czern. and Coss.]

An AFLP linkage map of Brassica juncea (L.) Czern and Coss was constructed using 88 recombinant inbred lines (RILs) from a cross between an Indian cultivar ‘Varuna’ and an accession from Poland ‘BEC-144’. The map included 91 AFLP markers organized on 19 linkage groups covering a total map distance of 1679.1 cM. A total of 14 QTLs were detected for oil content (2 QTLs), erucic acid (2 QTLs), eicosenoic acid (2 QTLs), linolenic acid (3 QTLs), linoleic acid (3 QTLs) and palmitic acid (2 QTLs). A specific genomic region on LG2 was associated with contents of three fatty acids: erucic acid, eicosenoic acid and linoleic acid. Some of the markers showed absolute linkage with the QTLs associated with the levels of linolenic acid, linoleic acid and oil content. These markers may be used for improvement of fatty acid profile of B. juncea.     

Evidence for Differential Resistance to Peronospora parasitica (downy mildew) in Accessions of Brassica juncea (mustard) at the Cotyledon Stage

Thirty-one Brassica juncea accessions were screened at the cotyledon stage for resistance to four isolates of Peronospora parasitica. Isolates R1 and P003 were derived from crops of oilseed rape (B. napus ssp. oleifera) in the UK and isolates IP01 and IP02 were derived from crops of mustard (B. juncea) in India. B. napus cv. Ariana, which was used as a susceptible control for isolates from B. napus, was resistant to isolates from B. juncea. All, B. juncea accessions were resistant to isolates from B. napus except one accession which expressed moderate resistance to isolate P003. Five groups of B. juncea accessions with differential resistance were identified. Lines homogeneous for resistance were selected from seedling populations of accessions that exhibited a heterogeneous reaction to isolates from B. juncea. The differential resistance identified in the B. juncea-P. parasitica combination can be used as a foundation for future studies of the genetics of the host-pathogen interaction and for breeding for disease resistance.     

Modification of erucic acid content in Indian mustard (Brassica juncea) by up-regulation and down-regulation of the Brassica juncea FAT TY ACID ELONGATION1 (BjFAE1) gene

In Brassicas, the Fatty Acid Elongation1 (FAE1) gene product, a 3-ketoacyl-CoA synthase, is the first in a 4-enzyme complex involved in the synthesis of erucic acid from oleic acid. The FAE1 homologue from Brassica juncea cv. Pusa Bold was cloned in a binary vector both in sense and antisense orientations under the control of the CaMV35S promoter. The recombinant binary vectors were used to transform B. juncea cv. RLM 198 via Agrobacterium tumefaciens. The presence of the transgene was confirmed by polymerase chain reaction and Southern hybridization. Northern and western analyses showed the expression of the gene and protein, respectively, in the transgenic plants. Analyses of the fatty acid profile of the seed oil from homozygous T4 generation seeds revealed that over-expression of the FAE1 gene caused a 36% increase in the percent of erucic acid (37–49% compared to 36% in untransformed control). The down-regulation of FAE1 caused an 86% decrease in the percent of erucic acid to as low as 5% in the seed oil of transgenic plants. Thus, it is clearly possible to alter erucic acid content of mustard by altering the expression level of the FAE 1 gene. S. Kanrar and J. Venkateswari equally contributed to this work.     

Mapping loci controlling the concentrations of erucic and linolenic acids in seed oil of Brassica napus L.

The quality of plant oil is determined by its component fatty acids. Relatively high levels of linolenic acid reduce the oxidative stability of the oil, and high levels of erucic acid in the diet have been associated with health problems. Thus, oilseed Brassica napus cultivars with low linolenic and low erucic acid contents are highly desirable for edible oil production. In order to identify genes controlling the levels of erucic and linolenic acids, we analyzed the oil composition of 99 F₁-derived doubled haploid lines from a cross between cv Major (high levels of erucic and linolenic acids) and cv Stellar (low levels of both fatty acids). A molecular marker linkage map of 199 loci for this population was used to identify quantitative trait loci (QTL) controlling oil composition. We identified two regions that accounted for nearly all of the phenotypic variation in erucic acid concentration and one region that accounted for 47% of the variation in linolenic acid concentration. The QTL associated with linolenic acid concentration mapped near a RFLP locus detected by a cDNA clone encoding an omega-3 desaturase, suggesting that the low linolenic acid content of Stellar may be due to a mutation in this gene.     

Characterization of disomic addition lines Brassica napus-Brassica nigra by isozyme,fatty acid,and RFLP markers

Summary Six Brassica napus — B. nigra disomic addition lines were characterized by isozyme, fatty acid, and RFLP markers. The markers were arranged in six synteny groups, representing six of the eight chromosomes present in the B. nigra genome. Synteny group 1 displayed high levels of linoleic and linolenic acids in the seeds of the B. nigra parent. Synteny group 3 accumulated higher levels of eicosenoic and erucic acid than B. nigra. Three of the lines transmitted the alien chromosome to 100% of the progeny. The rest had variable transmission rates but all were above 50%. Most of the lines produced disomic addition plants in their progeny, suggesting pollen transmission of the alien chromosome. In addition to the marked lines, six others remained unmarked. These could be grouped into two classes according to their alien chromosome transmission. It is likely that they represent the two other B. nigra chromosomes that remained uncharacterized by the markers. No diploid individuals carrying B. nigra genome-specific markers were detected in the progenies studied.     

Towards the genetic engineering of triacylglycerols of defined fatty acid composition: major changes in erucic acid content at the sn-2 position affected by the introduction of a 1-acyl-sn-glycerol-3-phosphate acyltransferase from Limnanthes douglasii into oil seed rape

A cDNA encoding a 1-acyl-sn-glycerol-3-phosphate acyltransferase from Limnanthes douglasii was introduced into oil seed rape (Brassica napus) under the control of a napin promoter. Seed triacylglycerols from transgenic plants were analysed by reversed-phase HPLC and trierucin was detected at a level of 0.4% and 2.8% in two transgenic plants but was not found in untransformed rape seed. Total fatty acid composition analysis of seeds from these selected plants revealed that the erucic acid content was no higher than the maximum found in the starting population. Analysis of fatty acids at the sn-2 position showed no erucic acid in untransformed rape but in the selected transgenic plants 9% (mol/mol) and 28.3% (mol/mol) erucic acid was present. These results conclusively demonstrate that the gene from L. douglasii encodes a 1-acyl-sn-glycerol-3-phosphate acyltransferase which can function in rape and incorporate erucic acid at the sn-2 position of triacylglycerols in seed. Additional modifications may further increase levels of trierucin.     

The role of glucosinolates and their hydrolysis products in oviposition and host-plant finding by cabbage webworm, Hellula undalis

The cabbage webworm, Hellula undalis (Fabricius) (Lepidoptera: Pyralidae), a tropical pest on crucifers (Brassicaceae), differentiated among host-plant species for oviposition in laboratory and field tests. White mustard, Sinapis alba (L.) var. Selinda, was the preferred host-plant, followed by Brassica juncea (L.) Czern. et. Coss var. Canadian brown mustard, and pak-choi, Brassica campestris L. ssp. chinensis var. Joi Choi, Black Behi and Bai Tsai. Glucosinolates (GS), secondary plant compounds characteristic to the Cruciferae plant family, and their breakdown products were analyzed by using HPLC and GC-MS-techniques. Species differed in GS composition and concentration. Content of GS was highest in S. alba with progressively lower contents detected in B. juncea and B. chinensis. The aromatic GS, 4-hydroxybenzyl-GS and benzyl-GS, were detected in S. alba. In B. juncea the alkenyl GS, allyl-GS, dominated, whereas in varieties of B. chinensis indolyl and alkenyl GS predominated. Oviposition of H. undalis females on the non-host-plant Vigna unguiculata ssp. sesquipedalis (L.) Fruwirth was stimulated by application of GS extracts from the crucifer species; the extract from S. alba was preferred, followed by extracts from B. juncea and B. chinensis. Hydrolysis of GS in the plant extract from B. chinensis causes loss of the oviposition stimulatory effect of the extract. Application of the GS, allyl-GS, and benzyl-GS also stimulated oviposition by H. undalis. Significantly more eggs were laid on leaves treated with the aromatic GS, benzyl-GS, than with the alkenyl GS, allyl-GS. Host-plant odor attracted H. undalis females but not males, in behavioral assays conducted in a Y-tube olfactometer. Low concentrations of the GS hydrolysis product, allyl-isothiocyanate, induced anemotaxis of females, but a high concentration of allyl-isothiocyanate was repellent. Oviposition by H. undalis females was not stimulated by host-plant volatiles. Females laid eggs on inserted traps and the walls of the Y-tube regardless of presence or absence of host-plant odor. The relevance of these results in the context of crucifer-insect interactions is discussed.