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Biotechnology of α-linolenic acid in oilseed rape (Brassica napus) using FAD2 and FAD3 from chia (Salvia hispanica)
XUE Yu-fei, INKABANGA TSEKE Alain, YIN Neng-wen, JIANG Jia-yi, ZHAO Yan-ping, LU Kun, LI Jia-na, DING Yan-song, ZHANG Shi-qing, CHAI You-rong
2023, 22 (12): 3810-3815.   DOI: 10.1016/j.jia.2023.05.018
Abstract123)      PDF in ScienceDirect      
α-Linolenic acid (ALA, 18:3Δ9,12,15) is an essential fatty acid for humans since it is the precursor for the biosynthesis of omega-3 long-chain polyunsaturated fatty acids (LC-PUFA). Modern people generally suffer from deficiency of ALA because most staple food oils are low or lack ALA content. Biotechnological enrichment of ALA in staple oil crops is a promising strategy. Chia (Salvia hispanica) has the highest ALA content in its seed oil among known oil crops. In this study, the FAD2 and FAD3 genes from chia were engineered into a staple oil crop, oilseed rape (Brassica napus), via Agrobaterium tumefaciens-mediated transformation of their LP4-2A fusion gene construct driven by the seed-specific promoter PNapA. In seeds of T0, T1, and T2 lines, the average ALA contents were 20.86, 23.54, and 24.92%, respectively, which were 2.21, 2.68, and 3.03 folds of the non-transformed controls (9.42, 8.78, and 8.22%), respectively. The highest seed ALA levels of T0, T1, and T2 plants were 38.41, 35.98, and 39.19% respectively, which were 4.10–4.77 folds of the respective controls. FA-pathway enzyme genes (BnACCD, BnFATA, BnSAD, BnSCD, BnDGAT1, BnDGAT2, and BnDGAT3) and positive regulatory genes (BnWRI1, BnLEC1, BnL1L, BnLEC2, BnABI3, BnbZIP67, and BnMYB96) were all significantly up-regulated. In contrast, BnTT1, BnTT2, BnTT8, BnTT16, BnTTG1, and BnTTG2, encoding negative oil accumulation regulators but positive secondary metabolism regulators, were all significantly down-regulated. This means the foreign ShFAD2-ShFAD3 fusion gene, directly and indirectly, remodeled both positive and negative loci of the whole FA-related network in transgenic B. napus seeds.
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Identification of quantitative trait loci and candidate genes controlling seed pigments of rapeseed
ZHU Mei-chen, HU Ran, ZHAO Hui-yan, TANG Yun-shan, SHI Xiang-tian, JIANG Hai-yan, ZHANG Zhi-yuan, FU Fu-you, XU Xin-fu, TANG Zhang-lin, LIU Lie-zhao, LU Kun, LI Jia-na, QU Cun-min
2021, 20 (11): 2862-2879.   DOI: 10.1016/S2095-3119(20)63377-9
Abstract162)      PDF in ScienceDirect      
Rapeseed (Brassica napus L.) is an important source of edible vegetable oil and feed protein; however, seed pigments affect the quality of rapeseed oil and the feed value of the residue from oil pressing.  Here, we used a population of rapeseed recombinant inbred lines (RILs) derived from the black-seeded male parent cultivar Zhongyou 821 and the yellow-seeded female parent line GH06 to map candidate genes controlling seed pigments in embryos and the seed coat.  We detected 94 quantitative trait loci (QTLs) for seed pigments (44 for embryos and 50 for seed coat), distributed over 15 of the 19 rapeseed chromosomes.  These included 28 QTLs for anthocyanidin content, explaining 2.41–44.66% of phenotypic variation; 24 QTLs for flavonoid content, explaining 2.41–20.26% of phenotypic variation; 16 QTLs for total phenol content, accounting for 2.74–23.68% of phenotypic variation; and 26 QTLs for melanin content, accounting for 2.37–24.82% of phenotypic variation, indicating that these traits are under multigenic control.  Consensus regions on chromosomes A06, A09 and C08 were associated with multiple seed pigment traits, including 15, 19 and 10 QTLs, respectively, most of which were major QTLs explaining >10% of the phenotypic variation.  Based on the annotation of the B. napus “Darmor-bzh” reference genome, 67 candidate genes were predicted from these consensus QTLs regions, and 12 candidate genes were identified as potentially involved in pigment accumulation by RNA-seq and qRT-PCR analysis.  These preliminary results provide insight into the genetic architecture of pigment biosynthesis and lay a foundation for exploring the molecular mechanisms underlying seed coat color in B. napus.
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Over-Expression of BnMAPK1 in Brassica napus Enhances Tolerance to Drought Stress
WENG Chang-mei, LU Jun-xing, WAN Hua-fang, WANG Shu-wen, WANG Zhen, LU Kun, LIANG Ying
2014, 13 (11): 2407-2415.   DOI: 10.1016/S2095-3119(13)60696-6
Abstract1257)      PDF in ScienceDirect      
Mitogen-activated protein kinases (MAPKs) are a family of Ser/Thr protein kinases widely conserved in all eukaryotes and involved in responses to biotic and abiotic stresses. In this study, two over-expressing BnMAPK1 oilseed rape lines, ov3 and ov11, were used to study the drought-resistant mechanism of BnMAPK1 under natural drought and simulation drought through spraying 10% PEG 8000 in seedlings. Zhongyou 821 (WT) was used as control. Compared with wild type, transgenic seedlings had higher leaf water content, higher root activity, slightly higher peroxidase (POD) and superoxide dismutase (SOD) activity, higher proline content and lower malondialdehyde (MDA) content. The expression of drought-resistant related genes, including P5CSB, PLC, LEA4 and SCE1, have been up-regulated in some degree and the expressed time of transgenic lines were earlier than that of wild type. These results suggested that over-expression of BnMAPK1 can enhance the resistance to drought in oilseed rape (Brassica napus).
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