A GmFAD3A mutation in the low linolenic acid soybean mutant C1640

The linolenic acid component is responsible for the flavour and odour instability of soybean oil. Thus, development of low linolenic acid soybean lines has been a major goal in crop science research. One such low linolenic acid soybean line, A5, contains a deletion of the omega‐3 fatty acid desaturase gene GmFAD3A. Another low linolenic acid soybean line, C1640, is allelic with A5, suggesting it too contains a mutation in the GmFAD3A gene. The purpose of this work was to characterize the genetic lesion in C1640 using GmFAD3A as a candidate gene. It could be that a premature stop codon is introduced in the gene, presumably rendering a non‐functional truncated enzyme. An assay to distinguish between the wild type and mutant alleles by using PCR followed by endonuclease digestion was developed. This assay will aid soybean breeders that are trying to incorporate the C1640 low linolenic acid trait into other lines.     

New mutants of winter rapeseed (Brassica napus L.) with changed fatty acid composition

Seeds of the winter oilseed rape (Brassica napus L.) line PN 3756/93 were treated with ethyl methanesulphonate to induce mutations in the fatty acid biosynthetic pathway. The seed mutagenic treatment was repeated in the M₂ generation. After treatments, individual seed and plant selections were made for changes in fatty acid composition during several generations of inbreeding. Self‐pollinated plants with changed fatty acid compositions were inbred to obtain genetically homozygous and stable mutant lines. Two mutants, M‐10453 and M‐10464, with increased levels of oleic acid (approximately 76%) and reduced linoleic and linolenic acid contents (8.5% and 7.5%, respectively) were selected. Gene or genes controlling desaturation of oleic acid were probably mutated in these plants. The third mutant, M‐681 had a very low linolenic acid content (approximately 2.6%) and increased linoleic acid content (approximately 26%). This would suggest the occurrence of mutations in genes controlling linoleic acid desaturation. The results of selection work during several generations showed that the environment had substantial influence on the composition of seed oil. This made the search for mutants with modify fatty acid compositions difficult. The induced mutants are not directly usable as new varieties, but can be used as parents in crosses for the development of high quality rapeseed varieties.     

Isolation of induced mutants in linseed (Linum usitatissimum) having reduced linolenic acid content

Summary Seeds of Linum usitatissimum cv. Glenelg were treated with either gamma-rays or EMS in an attempt to induce mutations with a lower level of linolenic acid in linseed oil. Two mutant lines were identified in which linolenic acid constituted approximately 29% of the total fatty acid content compared with 43% in seed oil from untreated Glenelg plants. The reduced level of linolenic acid in the mutants is accompanied by an increase in the level of linoleic acid to 30% compared with 18% in Glenelg, but there was no change in the proportions of other fatty acids. These proportions of linolenic acid and linolenic acid are respectively the highest and lowest yet reported in stable genotypes of L. usitatissimum. The strong inverse relationship between these two fatty acids in these genotypes suggests that linolenic acid is synthesised by desaturation of linolenic acid and indicates that it may be possible to breed an edible linseed oil having both low levels of linolenic acid and high levels of linolenic acid.     

Inheritance of reduced linolenic acid content in the Ethiopian mustard mutant N2-4961

The zero erucic acid Ethiopian mustard lines developed so far are characterized by an exceptionally high linolenic acid content in the seed oil. The mutant line N2‐4961, expressing low linolenic acid content in a high erucic acid background, was developed through chemical mutagenesis. The objective of this research was to study the inheritance of low linolenic acid content in this mutant. Line N2‐4961 was reciprocally crossed with its parent line C‐101 and the linolenic acid content of the reciprocal F₁, F₂ and BC₁ generations was studied. No maternal, cytoplasmic or dominance effects were detected in the analysis of F₁ seeds and F₁ plants from reciprocal crosses. Linolenic acid content segregated in 1: 2: 1 ratios in all the F₂ populations studied, suggesting monogenic inheritance. This was confirmed with the analysis of the reciprocal backcross generation. The simple inheritance of low linolenic acid content in N2‐4961 will facilitate the transference of this trait to zero erucic acid lines of Ethiopian mustard.     

Allelic variation in linolenic acid content of high erucic acid Ethiopian mustard and incorporation of the low linolenic acid trait into zero erucic acid germplasm

Zero erucic acid germplasm of Ethiopian mustard is characterized by high levels of linolenic acid (about 21%). Two genetic sources of low linolenic acid (N2‐4961 and HF‐186, about 5%), have been developed in a high erucic acid background. The objectives of this research were to study the genetic relationship between the two low linolenic acid lines and to transfer the trait to zero erucic acid germplasm. F1 seed generations from crosses between both lines had higher average linolenic acid concentration than both parents. F2 seeds segregated for linolenic acid content following a continuous variation from 1.8 to 7.4%, exceeding the limits of the parental distribution ranges. Transgressive recombinants with very low linolenic acid concentration (0.7‐2.7%) were confirmed in the F3 seed generation. The results suggested that N2‐4961 and HF‐186 possess alleles for low linolenic acid at different loci. Transgressive low linolenic acid F3 plants were crossed with plants of a zero erucic acid line and a selection for zero erucic, low linolenic acid was conducted. As a result, a zero erucic acid F3:4 line containing 1.5 ± 0.7% linolenic acid was developed.     

EMS Induction of Early Flowering Mutants in Spring Rape (Brassica napus)

Studies were undertaken to induce early flowering mutants by ethyl methanesulphonate (EMS) treatments of Brassica napus seeds. EMS treatments for 12 h of a highly inbred B. napus line TBS had adverse effects on M¹ plant development and fertility only when concentrations were greater than 1%. However, an EMS concentration of 1.5% did not reduce M₁ plant fertility to an extent which significantly reduced production of M₂ seeds. Genetic changes induced by EMS treatment and affecting flowering time were of three main types: (1) Changes within a polygenic system reflected by increased variation in flowering time among M₂ families. As the increase in variation was due primarily to a higher frequency of later flowering plants, these polygenic changes would be of little value in developing better-adapted cultivars. (2) Induction of a recessive mutation at a major gene locus which caused M₃ plants homozygous for the mutant gene to flower at least 20 days earlier than the parental line TB8. (3) Induction of a dominant mutation at a major gene locus which affects flowering time by causing a substantial reduction in vernalization requirement. M₂ plants carrying the mutant gene flowered as early as 59 days before the parental line. These major gene mutations could be rapidly exploited in the development of agronomically superior cultivars for short-season, lower rainfall environments in Western Australia.     

Selecting recombinants to stack high protein with high oleic acid and low linoleic acid in soybean (Glycine max)

Soybean accounts for over a quarter of the world's oilseed consumption and over 70% of the world's protein meal consumption. The separate development of high oleic, low linolenic acid (HOLL) soybean and high-protein (HP) soybean means that no soybean cultivar on the market has an optimal fatty acid profile and increased protein. The objective of this study was to develop and evaluate high protein, high oleic acid, and low linolenic acid (HP-HOLL) soybean. A five-gene stack was created using a two-phase forward breeding scheme and marker-assisted selection method. Forty-six HP-HOLL lines from three genetic backgrounds were grown in six environments in the Southeast United States. Although genotype-by-environment interaction was significant for seed composition traits, lines met the >75% and <3% cutoffs for oleic acid and linolenic acid, respectively, and met or exceeded the protein concentration of the HP parent. No negative interaction could be detected between the HP and HOLL traits. Additionally, yield testing in four environments indicated yield parity for some lines, suggesting HP and HOLL soybean cultivars with high yield could be selected.     

Identification of soybean genes related to fatty acid content based on a soybean genome collinearity analysis

Seed fatty acid content is an important consideration for soybean produced for food, feed, and industrial applications. In this study, MCScanX was used to analyze the entire soybean genome to generate a collinearity block, which was then used to assess the collinearity among the soybean fatty acid quantitative trait loci (QTL) in the SoyBase database. The hub‐QTLs located in the Gm06, Gm07, and Gm10 segments were identified. The Kyoto Encyclopedia of Genes and Genomes and gene ontology databases were used to analyze the genes in hub‐QTL regions, resulting in the identification of 17 candidate genes related to soybean fatty acid content. Two lines with different fatty acid contents and a recurrent parent were selected from a chromosome segment substitution line library for a subsequent quantitative real‐time polymerase chain reaction (qRT‐PCR) assay to verify the candidate gene expression patterns. Four genes were related to the total soybean fatty acid content, while three genes were related to the content of specific fatty acid types. The results of this study may be relevant for the fine mapping of soybean fatty acid QTLs/genes.     

Identifying and exploring significant genomic regions associated with soybean yield,seed fatty acids,protein and oil

Soybean [Glycine max (L.) Merrill] yield and seed fatty acids, protein, and oil content are important traits for which an improved understanding of significant genomic regions would be useful. To accomplish this, a soybean population consisting of 203 F5 derived recombinant inbred lines (RILs) was developed and genotyped with 11,633 polymorphic single nucleotide polymorphisms (SNPs). Each RIL was grown in a single plot at Knoxville, TN in 2010; followed by replicated, multi-location field trials in 2013 and 2014. The data from 2010, 2013, and 2014 were analyzed together in order to detect quantitative trait loci (QTL) for these traits, and 30 total QTLs were detected. Five QTLs are candidates for confirmed status and one QTL is a candidate for positional confirmation. Many of the genes with mutations in close proximity to the fatty acid QTLs are involved in biological processes for fatty acids and/or lipids and could be considered possible candidate genes. Similarly, genes with mutations in genomic regions near yield, protein, and oil QTLs were plentiful and may contribute to the variation observed in these traits. Except for yield and stearic acid, each trait displayed pleiotropic effects with other traits in this study. Notable are the pleiotropic effects for oleic and linolenic acid on chromosomes 9, 13, and 19. Overall, the findings from this research contribute new information to the genetic understanding of soybean yield and seed fatty acids, protein and oil content. This understanding will be useful in making trait improvements.     

大豆7S球蛋白α'亚基缺失新种质中黄608的分子鉴定

大豆7S球蛋白α'亚基含量与大豆的营养品质和加工特性关系密切。本研究利用聚丙烯酰胺凝胶电泳(SDS-PAGE)和免疫印迹法(Western blot)从中品661的EMS突变库中筛选出α'亚基缺失突变体中黄608。利用中黄608与登科1号创建了一个由210个个体组成的F2分离群体。遗传分析表明,α'亚基缺失由1对隐性单基因控制。利用连锁分析方法将该基因定位于第10染色体标记SSR10-1489与SSR10-1612之间,其中,包括控制α'亚基合成基因Cgy-1(Glyma.10G246300),序列分析发现,中黄608在Cgy-1第1外显子第84个碱基发生单碱基突变(G84→A84),导致氨基酸翻译提前终止。根据新发现的变异位点开发了共显性分子标记,并检测F2个体基因型,结果表明, Cgy-1基因型与α'亚基表型共分离。本研究不仅为大豆优质育种提供了新材料,同时也为分子育种提供了技术支持。     

EMS-induced mutation of an endoplasmic reticulum oleate desaturase gene (<Emphasis Type="Italic">FAD2</Emphasis>-<Emphasis Type="Italic">2</Emphasis>) results in elevated oleic acid content in rapeseed (<Emphasis Type="Italic">Brassica napus</Emphasis> L.)

The development of rapeseed cultivars (Brassica napus L.) with high oleic acid and low linolenic acid is highly desirable for food and industrial applications. In this study, the Korean rapeseed cultivar Tamla was used for ethyl methanesulfonate (EMS)-induced mutagenesis and seed oils were screened up to generation M₇ for high oleate mutants. Two mutant populations (M₇) with an average of approximately 76% oleic acid content were isolated. Yield components between two mutant populations and untreated Tamla plants were not substantially different, although the mutants in the vegetative stage were slightly smaller in size than Tamla. Genomic analyses of six fatty acid desaturase (four FAD2 and two FAD6) genes revealed that the elevated oleic acid content in the mutants is the result of single gene mutations. Changes in DNA sequence were observed in two genes out of six fatty acid desaturase (four FAD2 and two FAD6). FAD2-2 exhibited a 2-bp deletion in the upstream region of the gene in the two mutants, resulting in a severely truncated polypeptide (57 aa instead of 469 aa), while six point mutations in the other gene did not result in changes in the amino acid sequence. Based on these results, FAD2-2, an endoplasmic reticulum (ER) oleic acid desaturase, is affected in the mutants, resulting in a ~ 7% increase in oleic acid content in comparison to untreated Tamla plants. The induced mutants could be utilized for the development of high oleic oil rapeseed varieties and for regulatory studies of lipid metabolism in seed oils.     

EMS诱导小麦TcLr19感叶锈病突变体的筛选

为了利用小麦感叶锈病突变体进行抗病基因功能和机制研究,用EMS处理小麦抗叶锈病近等基因系Tc Lr19的种子,对获得的M2、M3植株进行农艺性状观察和田间抗叶锈性鉴定。结果显示,不同浓度的EMS溶液处理种子共获得367个M1单株,处理浓度为1.0%时,接近半致死剂量,且M2表型突变频率最高,适宜突变筛选。在2 359个M2突变群体中,共筛选到表型突变体38个,表型突变频率1.61%;筛选出感叶锈病突变体53个,感病突变频率2.25%。在459个M3感病突变群体中,共鉴定出146个感病突变体,其中M36-2、M333-8、M333-9、M333-11、M344-4、M396-8这6个M3感病突变材料,遗传稳定率较高,达70%以上。为保证结果的可靠性,试验中还利用Lr19的分子标记对突变体进行分子辅助筛选。结果表明,EMS诱变是获得小麦感叶锈病突变体的有效手段,不仅为小麦抗叶锈基因的克隆和功能研究提供了重要的基础材料,还为小麦育种提供了新的种质。     

对一个新发现的棉纤维突变体的鉴定及特性分析

在农杆菌介导的转基因组织培养再生后代中发现了一个无绒有絮的纤维发育突变体,通过自交选择T₃代获得其纯合体,命名为CM突变体。尽管CM突变体从转基因后代中发现,但和转基因插入位点无关,推测是在组织培养过程中产生的点突变所致。通过与陆地棉遗传标准系TM-1,海岛棉军海1号,以及新乡小吉无绒有絮(XinFLM),新乡小吉无绒无絮(XinWX),徐州142无绒无絮(XZ142WX),显性光子N1N1,隐性光子n2n2、SL-7-1、MD17及T586等一系列纤维发育突变体分别配制F₂组合进行突变基因的遗传及等位性分析,结果表明CM突变体与纤维发育正常的TM-1和军海1号杂交,F₁表型均为无绒有絮,F₂表现无绒有絮和有绒有絮3∶1分离,说明该突变体与纤维发育正常材料相比,在短绒发育方面存在一个位点的差异,该突变性状由单显性基因控制。等位性测验及分子定位均表明, 控制该突变体短绒的基因与控制N1N1显性光子的N1基因等位。扫描电镜进一步证明该基因突变会造成纤维起始突起延迟。与N1N1突变体相比,CM突变体的衣分比N1N1显著高,而百粒重比N1N1极显著低。推测CM突变体中的突变基因与显性N1基因为复等位基因。     

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.     

Inheritance of the four-seeded-pod trait in a soybean mutant and marker-assisted selection for this trait

A high seeds‐per‐pod value is a crucial component of soybean seed yield, but reliable information is lacking on the inheritance and early selection for the four‐seeded pod (4SP) trait. The inheritance of 4SP was followed in crosses originating from an EMS‐derived mutant line (E182, 15% 4SP) with its parental cultivar, ‘Ludou No. 4’ (no 4SP), and the efficiencies of both a molecular and morphological marker selection were determined. The plants of two F₂ populations (with E182 as one of the parents) showed a segregation ratio of 3 : 1 (low to high SP values), indicating a single recessive locus; this mode of inheritance was confirmed by subsequent analysis of the F_{2 : 3} families. Among four microsatellite markers linked with the 4SP trait, Sat_107 was the closest to the 4SP locus (3.2 ± 1.11 cM). Both the molecular marker Sat_107 and the morphological marker, narrow leaflet (NL) associated with the 4SP locus were effective in selecting the 4SP trait, although with different efficiencies. The molecular marker was efficient in selecting soybean progenies from the cross in which one parent was the E182 mutant line because it was tightly linked to the mutated 4SP locus. In the other cross, with 4SP not derived from E182, the efficiency was reduced in comparison with the morphological marker.     

Identification of RAPD markers linked to linolenic acid genes in rapessed

Summary The inheritance of the low linolenic acid content (derivated from mutant lines) in rapeseed was investigated. Molecular techniques of gene mapping through RAPD markers were applied on a microspore-derived progeny from a high × low linolenic acid F₁ hybrid. Bulked segregant analysis made it possible to test rapidly number of RAPD primers. Two linkage groups of 6 markers (72.7 cM and 75.6 cM) were determined. Each corresponded to a major QTL which explained 24% and 30.7% of the total phenotypic variation of the linolenic acid content. It was confirmed that two independant mutations were implied in the low linonenic acid content.     

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.     

高油酸玉米突变体的诱导和遗传分析

油酸,亚油酸及饱和脂肪酸含量决定玉米油的品质,研究结果表明,利用ＥＭＳ花粉诱变技术,可获得高油酸等玉米突变体,高油酸含量的遗传受部分显性生效基因控制,基因的显性方向指向增效,但是,在同一基因位点上,可能受一些微恢复等位基因累加作用的影响,卡平方检验符合１对部分显性基因的遗传规律,与同类研究结果基本一致,优质油玉米育种,不仅要提高玉米的含油量,还应提高油分中的油酸含量,降低饱和脂肪酸含量,改善玉米的