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沙棘(Hippophae rhamnoides L.)是胡颓子科沙棘属的一种雌雄异株的灌木或小乔木,耐旱性和耐寒性良好。沙棘果实具有独特的药用和营养价值,在中国和俄罗斯有上百年的药用历史[1]。沙棘果实富含维生素、有机酸、氨基酸、脂肪酸、抗氧化剂和类黄酮[2-4],在炎症性疾病、肝脏性疾病、动脉粥样硬化等疾病的治疗以及保护心脏等方面具有显著功效[5]。在沙棘种子含有的众多脂肪酸中,α-亚麻酸(α-Linolenic acid,C18:3N3)的含量十分丰富,且α-亚麻酸对大脑发育、心血管健康、炎症治疗等都具有重要作用[6-9],但α-亚麻酸是人体必需脂肪酸,不能在人体内合成,只能通过食物摄取。豆油、花生油等常见植物食用油中的主要脂肪酸为油酸、亚油酸、棕榈酸和硬脂酸[10-11],α-亚麻酸含量极低,而沙棘中高积累20.3%~36.3%的α-亚麻酸[12],并且沙棘作为耐旱木本植物可以在沙地、荒山、盐碱地等边际土地生长,可避免与大田油料作物争夺土地,是理想的α-亚麻酸供给对象。
目前,油料作物脂肪酸合成途径及调控基因的研究较多,在沙棘的相关研究中多围绕脂肪酸组分及脂肪酸测定方法展开,少有针对沙棘中α-亚麻酸合成机制的研究。本研究选取α-亚麻酸含量存在差异的2个沙棘品种种子,对其含量差异及相关基因进行分析,在基因表达水平上研究沙棘中α-亚麻酸合成及代谢规律,为培育高α-亚麻酸含量的沙棘良种奠定理论基础。
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利用气相色谱分析,在2个品种沙棘的种子中共检测到27种脂肪酸(表1),其中,主要脂肪酸为C18:3N3(α-亚麻酸)、C18:2(亚油酸)、C16:0(棕榈酸)、C18:1(油酸)、C18:0(硬脂酸)。随着果实不断成熟,2个品种沙棘种子中,5种主要脂肪酸的含量均逐渐增加,其中,α-亚麻酸和亚油酸含量上升幅度较大。
表 1 XY与FN 2个沙棘品种种子在不同发育时期脂肪酸含量的动态变化
Table 1. Dynamic changes of fatty acid composition in the seeds of XY and FN at different developmental stages
μg·g−1 组分
CompositionsXY FN T1 T2 T3 T1 T2 T3 C10:0 1.04±0.09 b 1.75±0.13 ab 2.43±0.69 a 1.04±0.07 b 1.72±0.20 ab 2.24±0.20 a C12:0 1.11±0.27 a 2.06±0.78 a 5.62±0.32 a 0.34±0.40 a 1.34±0.17 a 2.09±1.25 a C13:0 0.08±0.05 b 0.30±0.08 ab 0.35±0.188 a 0.08±0.02 b 0.31±0.10 ab 0.48±0.10 a C14:0 20.70±2.32 c 40.23±2.638 a 53.98±18.17 a 17.66±1.34 c 40.85±7.66 a 58.91±7.54 a C14:1 136.34±14.20 a 145.86±15.94 a 122.48±14.74 a 121.50±9.22 a 121.08±9.35 a 117.06±7.36 a C15:0 6.20±1.43 c 19.42±1.16 b 28.45±10.91 ab 6.84±0.91 c 24.40±5.20 b 37.00±5.97 a C15:1 16.90±2.09 a 14.37±2.21 ab 8.59±0.85 c 14.65±0.78 ab 12.06±4.19 abc 10.52±1.83 bc C16:0 1 219.70±135.84 b 2 337.41±132.23 a 2 873.02±915.18 a 1 141.08±69.79 b 2 482.63±282.95 a 3 165.25±346.54 a C16:1 38.04±6.73 c 70.55±12.28 bc 112.18±36.82 ab 28.40±4.84 c 86.75±22.13 ab 126.20±25.12 a C17:0 7.89±1.39 b 16.93±1.34 a 19.21±6.58 a 9.02±0.85 b 15.92±1.62 a 21.51±2.86 a C17:1 66.01±7.57 a 51.99±6.67 ab 47.64±2.13 ab 56.48±12.46 ab 44.60±3.13 b 51.85±7.10 ab C18:0 695.59±62.14 c 1 330.90±97.82 ab 1 656.18±553.21 a 634.31±23.71 c 1 035.51±73.38 bc 1 280.61±137.68 ab C18:1 78.99±20.21 d 1 824.10±207.78 c 3 447.79±1 190.81 a 101.66±20.47 d 2 197.36±133.31 bc 3 029.64±464.58 ab C18:2 311.23±62.81 d 4 231.52±246.51 c 6 879.55±2 327.19 ab 265.09±48.17 d 5 028.36±863.33 bc 7 443.04±795.32 a C18:3N3 298.54±76.45 c 4 980.34±261.85 ab 7 463.53±2 636.16 a 223.36±46.56 c 4 307.42±690.29 b 6 526.07±969.49 ab C20:0 19.50±3.83 c 102.61±13.30 b 200.82±74.68 a 16.14±2.06 c 94.96±7.90 b 139.33±19.39 b C20:1 7.42±1.60 c 50.74±4.18 b 90.17±30.25 a 5.87±0.58 c 53.50±4.71 b 83.41±16.89 a C20:2 0.93±0.22 c 8.69±0.52 b 14.62±5.51 ab 0.81±0.13 c 9.72±3.29 b 16.89±3.08 a C21:0 3.85±0.72 b 8.44±0.77 ab 11.93±3.97 a 3.76±0.51 b 8.15±1.48 ab 11.94±2.08 a C20:3N3 1.90±0.21 bc 3.02±0.23 ab 3.86±0.93 a 1.53±0.28 c 2.51±0.55 bc 3.94±0.72 a C20:5N3 0 a 0.15±0.08 a 1.04±0.79 a 0 a 0.33±0.03 a 0.87±0.61 a C22:0 12.20±1.62 c 36.59±4.23 b 57.26±20.45 ab 14.29±2.13 c 44.80±9.54 b 68.61±10.83 a C22:1N9 24.60±3.00 ab 29.29±6.16 a 26.30±5.35 ab 16.34±0.58 b 21.45±3.73 ab 20.85±4.84 ab C23:0 2.00±0.44 c 10.73±1.28 b 18.25±7.01 a 2.41±0.44 c 9.76±2.18 b 15.84±3.27 ab C24:0 4.74±0.60 c 17.42±2.00 b 29.66±10.75 a 5.35±0.46 c 18.09±3.08 b 26.69±4.54 ab C24:1 4.05±3.29 a 11.48±0.91 a 6.68±3.95 a 4.38±3.87 a 3.94±3.01 a 4.47±1.55 a C22:6N3 2.62±0.11 c 3.43±0.05 a 3.25±0.25 ab 2.55±0.08 c 3.07±0.09 b 3.26±0.18.00 ab 注:同行不同字母代表显著差异(P< 0.05)。
Note: Different letters in the same line represent significant differences(P< 0.05)从图1可以看出:XY与FN的5种主要脂肪酸的含量均在T2时期表现出显著差异。按照脂肪酸含量从大到小排序,在XY中,依次为α-亚麻酸、亚油酸、油酸、棕榈酸、硬脂酸,含量最高的脂肪酸为α-亚麻酸,亚油酸次之;而在FN中,脂肪酸从大到小排序依次为亚油酸、α-亚麻酸、棕榈酸、油酸、硬脂酸,亚油酸大于α-亚麻酸含量(图1)。
图 1 沙棘XY与FN种子中主要脂肪酸组分的含量变化
Figure 1. Distribution of main fatty acid components in XY and FN seeds of sea buckthorn
在T2时期,2个沙棘品种种子中α-亚麻酸与亚油酸的比值(α-亚麻酸/亚油酸)存在显著差异(P<0.05)。XY中,T2时期3个样本的α-亚麻酸/亚油酸比值分别为1.21、1.17、1.16,而FN分别为0.92、0.75、0.92(表2)。
表 2 T2时期XY与FN种子中的C18:3N3/C18:2
Table 2. C18:3n3 / C18:2 in XY and FN seeds of sea buckthorn at T2
品种
CultivarC18:3N3/C18:2 T2 T2-1 T2-2 T2-3 XY 1.182a 1.21 1.17 1.16 FN 0.862b 0.92 0.75 0.92 注:不同字母代表显著差异(P<0.05),T2-1、T2-2、T2-3代表T2时期的3个生物学重复样本。
Notes: Different letters in the same column represent significant differences(P<0.05).T2-1、T2-2 and T2-3represent three biological replicates of the T2 period. -
以XY与FN种子的3个发育时期、每个发育时期3个生物学重复的18个样品为材料,通过RNA-seq测序分析共产生140 G clean data,933 483 428条高质量序列(Clean reads)(Q20>97%;Q30>93%),其中,81.5%的序列唯一比对到参考基因组上。对2个品种3个不同发育时期及相同时期不同品种间的表达基因进行差异分析,共鉴定出24 917个差异表达基因(DEGs)。从图2A样本DEGs的主成分分析(PCA)可以看出:2个沙棘品种各个时期生物学重复聚类效果良好,不同时期之间与不同品种之间数据离散效果良好。通过DEGs的分布图(图2B)可以看出:在T1-T3的发育过程中,在T2时期XY与FN之间的DEGs最多。
依据序列同源性将24 917个差异表达基因进行GO功能注释,2 605个基因注释为生物过程(Biological process),554个基因注释为细胞组成(Cellular component),1 354个基因注释为分子功能(Molecular function)。为了进一步了解基因的生物学功能和相互作用,使用KEGG数据库对基因进行通路富集分析。24 917个DEGs注释到122个KEGG通路,其中,有94个DEGs的功能富集于脂肪酸合成(Fatty acid biosynthesis)通路、脂肪酸代谢(Fatty acid metabolism)通路、α-亚麻酸代谢(α-Linolenicacid metabolism)通路和亚油酸代谢(Linoleicacid metabolism)通路。
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T2时期,基因FAD2在2个沙棘品种中的表达量差异显著(图3),在XY中的表达量显著高于FN,是FN的3.83倍,为α-亚麻酸(C18:3N3)的合成提供充足底物。FAD2基因在XY和FN中表达量均为先上升后下降,在第2时期表达量急剧上升达到峰值,与XY和FN中的亚油酸(C18:2)含量在第二时期显著增加(图1)相一致。
图 3 沙棘种子发育过程中与α-亚麻酸合成相关的基因表达模式
Figure 3. Gene expression profiles of Alpha linolenic acid biosynthetic genes during seed development of sea buckthorn
基因FAD3和基因FAD7调控亚麻酸生成α-亚麻酸(图4),在FN中的整体表达水平远低于XY,与XY中α-亚麻酸含量高于FN(图1)的结果相一致。基因FAD3在FN种子中的表达呈下调趋势,在XY种子中的表达水平先急剧上升,在T2时期达到峰值,后在T3时期表达量下调;T1时期,基因FAD3在XY中的表达量是FN 中的2.02倍,T2时期基因FAD3在XY和FN中的表达差异显著,在XY中的表达量是FN中的13.63倍(图3)。基因FAD7在XY和FN中的表达趋势均为先上升后下降,T1和T2时期在XY中的表达量显著高于在FN中的表达量,T1时期在XY中的表达量是FN 中的2.09倍,T2时期在XY中的表达量是FN 中的1.72倍(图3)。
图 4 与α-亚麻酸生物合成与代谢相关的途径
Figure 4. Pathway related to the biosynthesis and metabolism of α-linolenic acid
基因LOX调控α-亚麻酸氧化(图4),生成脂肪酸氢过氧化物,是调控α-亚麻酸转化为其他物质的第一步,基因LOX3.1在XY和FN种子的生长发育过程中表达趋势均为逐渐下降,在FN中的表达量高于XY,T1时期基因LOX3.1在FN中的表达量是XY中的3.58倍,T2时期基因LOX3.1在FN中的表达量是XY中的8.02倍(图3)。基因LOX3.2在FN中的表达量同样高于XY,T1时期在FN中的表达量是XY中的3.24倍,T2时期在FN中的表达量是XY中的7.12倍(图3)。基因LOX的高表达不利于α-亚麻酸的积累,与图1中所显示的XY中α-亚麻酸含量高于FN的结果相一致。
不同沙棘品种种子中α-亚麻酸含量差异及相关基因分析
Analysis of α-Linolenic Acid Content and Related Genes in Seeds of Sea Buckthorn Cultivars
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摘要:
目的 探讨α-亚麻酸含量存在差异的2个沙棘品种种子中的基因调控差异,为进一步提高沙棘种子中α-亚麻酸含量、培育沙棘良种奠定基础。 方法 以2个不同品种的沙棘种子为研究对象,通过气相色谱-质谱分析和转录组测序分析确定2个沙棘品种种子中的α-亚麻酸含量差异以及相关差异表达基因。 结果 蒙古大果沙棘(Hippophae rhamnoides ‘Mongolia’向阳)(XY)和中国沙棘(H. rhamnoides ‘Sinensis’丰宁)(FN)种子中的α-亚麻酸含量占比不同,XY中含量最高的脂肪酸是α-亚麻酸,其次是亚油酸,而FN中α-亚麻酸含量次于亚油酸。半成熟期(T2),基因FAD2和基因FAD3在XY中的表达量均显著高于FN,分别是FN的3.83倍和13.63倍。未成熟期(T1)和T2时期基因FAD7在XY中的表达量均显著高于在FN中的表达量,分别是FN的2.09倍和1.72倍。相反,T2时期基因LOX3.1和LOX3.2在FN中的表达量均高于XY,分别是XY的8.02倍和7.12倍。 结论 沙棘种子中α-亚麻酸的高积累源于多个基因的协同作用,基因FAD2、FAD3、FAD7的高表达和基因LOX3.1和LOX3.2的低表达共同调控α-亚麻酸的高积累。 Abstract:Objective To study the differences of gene regulation in seeds of two sea buckthorn (Hippophae rhamnoides) cultivars with different α-linolenic acid content and to find the method of further increase the content of α-linolenic acid in H. rhamnoides seed. Method The difference in α-linolenic acid content and the expressed genes in the seeds of two H. rhamnoides cultivars were analyzed and determined by gas chromatography-mass spectrometry and transcriptomics. Result The proportion of α-linolenic acid in H. rhamnoides 'Mongolia' (XY) and H. rhamnoides ‘Sinensis’ (FN) seeds was different. The fatty acid with the highest content of XY was α-linolenic acid, followed by linoleic acid. The content of α-linolenic acid in FN was lower than that of linoleic acid. In semi-mature stage (T2), the expression levels of gene FAD2 and gene FAD3 in XY were 3.83 times and 13.63 times that in FN respectively. The expression of gene FAD7 in XY was 2.09 times and 1.72 times that in FN in immature stage (T1) and T2respectively. It was consistent with the trend of linoleic acid and α-linolenic acid content. On the contrary, the expressions of gene LOX3.1 and gene LOX3.2 in FN were 8.02 times and 7.12 times that of XY in T2 respectively. Conclusion The high accumulation of α-linolenic acid in H. rhamnoides seeds is due to the synergistic effect of multiple genes. High expression of gene FAD2, FAD3, FAD7 and low expression of gene LOX3.1, LOX3.2 contribute to the accumulation of α-linolenic acid. -
表 1 XY与FN 2个沙棘品种种子在不同发育时期脂肪酸含量的动态变化
Table 1. Dynamic changes of fatty acid composition in the seeds of XY and FN at different developmental stages
μg·g−1 组分
CompositionsXY FN T1 T2 T3 T1 T2 T3 C10:0 1.04±0.09 b 1.75±0.13 ab 2.43±0.69 a 1.04±0.07 b 1.72±0.20 ab 2.24±0.20 a C12:0 1.11±0.27 a 2.06±0.78 a 5.62±0.32 a 0.34±0.40 a 1.34±0.17 a 2.09±1.25 a C13:0 0.08±0.05 b 0.30±0.08 ab 0.35±0.188 a 0.08±0.02 b 0.31±0.10 ab 0.48±0.10 a C14:0 20.70±2.32 c 40.23±2.638 a 53.98±18.17 a 17.66±1.34 c 40.85±7.66 a 58.91±7.54 a C14:1 136.34±14.20 a 145.86±15.94 a 122.48±14.74 a 121.50±9.22 a 121.08±9.35 a 117.06±7.36 a C15:0 6.20±1.43 c 19.42±1.16 b 28.45±10.91 ab 6.84±0.91 c 24.40±5.20 b 37.00±5.97 a C15:1 16.90±2.09 a 14.37±2.21 ab 8.59±0.85 c 14.65±0.78 ab 12.06±4.19 abc 10.52±1.83 bc C16:0 1 219.70±135.84 b 2 337.41±132.23 a 2 873.02±915.18 a 1 141.08±69.79 b 2 482.63±282.95 a 3 165.25±346.54 a C16:1 38.04±6.73 c 70.55±12.28 bc 112.18±36.82 ab 28.40±4.84 c 86.75±22.13 ab 126.20±25.12 a C17:0 7.89±1.39 b 16.93±1.34 a 19.21±6.58 a 9.02±0.85 b 15.92±1.62 a 21.51±2.86 a C17:1 66.01±7.57 a 51.99±6.67 ab 47.64±2.13 ab 56.48±12.46 ab 44.60±3.13 b 51.85±7.10 ab C18:0 695.59±62.14 c 1 330.90±97.82 ab 1 656.18±553.21 a 634.31±23.71 c 1 035.51±73.38 bc 1 280.61±137.68 ab C18:1 78.99±20.21 d 1 824.10±207.78 c 3 447.79±1 190.81 a 101.66±20.47 d 2 197.36±133.31 bc 3 029.64±464.58 ab C18:2 311.23±62.81 d 4 231.52±246.51 c 6 879.55±2 327.19 ab 265.09±48.17 d 5 028.36±863.33 bc 7 443.04±795.32 a C18:3N3 298.54±76.45 c 4 980.34±261.85 ab 7 463.53±2 636.16 a 223.36±46.56 c 4 307.42±690.29 b 6 526.07±969.49 ab C20:0 19.50±3.83 c 102.61±13.30 b 200.82±74.68 a 16.14±2.06 c 94.96±7.90 b 139.33±19.39 b C20:1 7.42±1.60 c 50.74±4.18 b 90.17±30.25 a 5.87±0.58 c 53.50±4.71 b 83.41±16.89 a C20:2 0.93±0.22 c 8.69±0.52 b 14.62±5.51 ab 0.81±0.13 c 9.72±3.29 b 16.89±3.08 a C21:0 3.85±0.72 b 8.44±0.77 ab 11.93±3.97 a 3.76±0.51 b 8.15±1.48 ab 11.94±2.08 a C20:3N3 1.90±0.21 bc 3.02±0.23 ab 3.86±0.93 a 1.53±0.28 c 2.51±0.55 bc 3.94±0.72 a C20:5N3 0 a 0.15±0.08 a 1.04±0.79 a 0 a 0.33±0.03 a 0.87±0.61 a C22:0 12.20±1.62 c 36.59±4.23 b 57.26±20.45 ab 14.29±2.13 c 44.80±9.54 b 68.61±10.83 a C22:1N9 24.60±3.00 ab 29.29±6.16 a 26.30±5.35 ab 16.34±0.58 b 21.45±3.73 ab 20.85±4.84 ab C23:0 2.00±0.44 c 10.73±1.28 b 18.25±7.01 a 2.41±0.44 c 9.76±2.18 b 15.84±3.27 ab C24:0 4.74±0.60 c 17.42±2.00 b 29.66±10.75 a 5.35±0.46 c 18.09±3.08 b 26.69±4.54 ab C24:1 4.05±3.29 a 11.48±0.91 a 6.68±3.95 a 4.38±3.87 a 3.94±3.01 a 4.47±1.55 a C22:6N3 2.62±0.11 c 3.43±0.05 a 3.25±0.25 ab 2.55±0.08 c 3.07±0.09 b 3.26±0.18.00 ab 注:同行不同字母代表显著差异(P< 0.05)。
Note: Different letters in the same line represent significant differences(P< 0.05)表 2 T2时期XY与FN种子中的C18:3N3/C18:2
Table 2. C18:3n3 / C18:2 in XY and FN seeds of sea buckthorn at T2
品种
CultivarC18:3N3/C18:2 T2 T2-1 T2-2 T2-3 XY 1.182a 1.21 1.17 1.16 FN 0.862b 0.92 0.75 0.92 注:不同字母代表显著差异(P<0.05),T2-1、T2-2、T2-3代表T2时期的3个生物学重复样本。
Notes: Different letters in the same column represent significant differences(P<0.05).T2-1、T2-2 and T2-3represent three biological replicates of the T2 period. -
[1] 臧茜茜, 邓乾春, 从仁怀, 等. 沙棘油功效成分及药理功能研究进展[J]. 中国油脂, 2015, 40(5):76-81. doi: 10.3969/j.issn.1003-7969.2015.05.017 [2] Tiitinen K M, Hakala M A, Kallio H P. Quality components of Sea buckthorn (<italic>Hippophae rhamnoides</italic>) varieties[J]. J Agric Food Chem, 2005, 53(5): 1692-1699. doi: 10.1021/jf0484125 [3] Tiitinen K M, Yang B, Haraldsson G G,<italic> et al</italic>. Fast analysis of sugars, fruit acids, and vitamin C in Sea buckthorn (<italic>Hippophae rhamnoides</italic> L.) varieties[J]. J Agric Food Chem, 2006, 54(7): 2508-2513. doi: 10.1021/jf053177r [4] Andersson S C, Olsson M E, Johansson E,<italic> et al</italic>. Carotenoids in Sea buckthorn (<italic>Hippophae rhamnoides</italic> L.) berries during ripening and use of pheophytin a as a maturity marker[J]. Journal of Agricultural & Food Chemistry, 2009, 57(1): 250-258. [5] Kanayama Y, Kato K, Stobdan T,<italic> et al</italic>. Research progress on the medicinal and nutritional properties of Sea buckthorn (<italic>Hippophae rhamnoides</italic>)-a review[J]. Journal of Horticultural Science and Biotechnology, 2012, 87(3): 203-210. doi: 10.1080/14620316.2012.11512853 [6] Marta S M, Cuenca A P. The impact of Sea buckthorn oil fatty acids on human health[J]. Lipids in Health and Disease, 2019, 18(1): 145-157. doi: 10.1186/s12944-019-1065-9 [7] Das U. Essential fatty acids - a review[J]. Current Pharmaceutical Biotechnology, 2006, 7: 467-482. doi: 10.2174/138920106779116856 [8] Leaf A. Omega-3 fatty acids and prevention of arrhythmias[J]. Current Opinion in Lipidology, 2007, 18(1): 31-34. doi: 10.1097/MOL.0b013e328012d61b [9] Wang C, Harris W S, Mei C,<italic> et al</italic>. n 3 Fatty acids from fish or fish-oil supplements, but not -linolenic acid, benefit cardiovascular disease outcomes in primary- and secondary-prevention[J]. Am J Clin Nutr, 2006, 84(1): 5-17. doi: 10.1093/ajcn/84.1.5 [10] 姚振纯, 刘继德. 大豆脂肪酸组分与改良[J]. 大豆通报, 1997(1):14-14. [11] 迟晓元, 郝翠翠, 潘丽娟, 等. 不同花生品种脂肪酸组成及其积累规律的研究[J]. 花生学报, 2016, 45(3):32-36. [12] Yang B, Kallio H P. Fatty acid composition of lipids in Sea buckthorn (<italic>Hippophae rhamnoides</italic> L.) berries of different origins[J]. Journal of Agricultural & Food Chemistry, 2001, 49: 1939-1947. [13] Ingrid E J T, Chanderbali A S, Gitzendanner M A,<italic> et al</italic>. Modified CTAB and TRIzol protocols improve RNA extraction from chemically complex embryophyta[J]. Applications in Plant Sciences, 2015, 3(5): 1400105. doi: 10.3732/apps.1400105 [14] Wang L, Feng Z, Wang X,<italic> et al</italic>. DEGseq: An R package for identifying differentially expressed genes from RNA-seq data[J]. Bioinformatics, 2009, 26: 136-138. [15] Young M D, Wakefield M J, Smyth G K,<italic> et al</italic>. Gene ontology analysis for RNA-seq: accounting for selection bias[J]. Genome biology, 2010, 11(2): 1-12. [16] Mao X, Cai T, Olyarchuk JG,<italic> et al</italic>. Automated genome annotation and pathway identification using the KEGG Orthology (KO) as a controlled vocabulary[J]. Bioinformatics, 2005, 21(19): 3787-3793. doi: 10.1093/bioinformatics/bti430 [17] Jadhav A, Katavic V, Marillia E F,<italic> et al</italic>. Increased levels of erucic acid in <italic>Brassica carinata</italic> by co-suppression and antisense repression of the endogenous FAD2 gene[J]. Metabolic Engineering, 2005, 7(3): 215-220. doi: 10.1016/j.ymben.2005.02.003 [18] Niu B, Guo L, Zhao M,<italic> et al</italic>. Molecular cloning, characterization, and expression of an ω-3 fatty acid desaturase gene from <italic>Sapium sebiferum</italic>[J]. Journal of Bioscience & Bioengineering, 2008, 106(4): 375-380. [19] DellaPenna, D. Plant metabolic engineering[J]. Plant Physiology, 2001, 125(1): 160-163. doi: 10.1104/pp.125.1.160 [20] Huang J, Zhang T, Zhang Q,<italic> et al</italic>. The mechanism of high contents of oil and oleic acid revealed by transcriptomic and lipidomic analysis during embryogenesis in <italic>Carya cathayensis </italic>Sarg<italic></italic>[J]. BMC Genomics, 2016, 17(1): 113-131. doi: 10.1186/s12864-016-2434-7 [21] 李 丽, 崔顺立, 穆国俊, 等. 高油酸花生遗传改良研究进展[J]. 中国油料作物学报, 2019, 41(6):986-997. [22] Liang L H, Jie Y Z, Li X,<italic> et al</italic>. Identification and evaluation of ω-3 fatty acid desaturase genes for hyperfortifying α-linolenic acid in transgenic rice seed[J]. Journal of Experimental Botany, 2012, 63(8): 3279-3287. doi: 10.1093/jxb/ers051 [23] Hiroaki K, Tatsurou H, Gorou H,<italic> et al</italic>. Genetic enhancement of cold tolerance by expression of a gene for chloroplast ω-3 fatty acid desaturase in transgenic tobacco[J]. Plant Physiology, 1994, 105: 601-605. doi: 10.1104/pp.105.2.601 [24] 韩 平, 阮成江, 丁 健, 等. 紫斑牡丹种子高积累碳十八不饱和脂肪酸的多基因调控[J]. 分子植物育种, 2019, 17(7):2101-2108. [25] Bhunia R K, Chakraborty A, Kaur R,<italic> et al</italic>. Seed-specific increased expression of 2S albuminpromoter of sesame qualifies it as a useful genetic tool for fatty acid metabolic engineering and related transgenic intervention in sesame and other oil seed crops[J]. Plant Molecular Biology, 2014, 86: 351-365. doi: 10.1007/s11103-014-0233-6 [26] Wu P, Zhang S, Zhang L,<italic> et al</italic>. Functional characterization of two microsomal fatty acid desaturases from <italic>Jatropha curcas</italic> L.[J]. Journal of Plant Physiology, 2013, 170(15): 1360-1366. doi: 10.1016/j.jplph.2013.04.019 [27] Kühn H, Borchert A. Regulation of enzymatic lipid peroxidation: the interplay of peroxidizing and peroxide reducing enzymes[J]. Free Radical Biology and Medicine, 2002, 33(2): 154-172. doi: 10.1016/S0891-5849(02)00855-9 [28] 陈 昊, 谭晓风. 基于油脂合成期油桐种仁转录组数据的α-亚麻酸代谢途径解析[J]. 林业科学, 2015, 51(3):41-48. [29] Xie D, Dai Z, Yang Z,<italic> et al</italic>. Combined genome-wide association analysis and transcriptome sequencing to identify candidate genes for flax seed fatty acid metabolism[J]. Plant science: an international journal of experimental plant biology, 2019, 286: 98-107.