作物学报 ›› 2017, Vol. 43 ›› Issue (11): 1588-1595.doi: 10.3724/SP.J.1006.2017.01588
杨静,邢国杰,牛陆,贺红利,杜茜,郭东全,袁英*,杨向东*
YANG Jing,XING Guo-Jie,NIU Lu,HE Hong-Li,DU Qian,GUO Dong-Quan,YUAN Ying*,YANG Xiang-Dong*
摘要:
油酸含量是评价大豆油食用品质和稳定性的重要指标。本研究采用农杆菌介导转化法,将反义GmFAD2-1B基因导入栽培大豆品种,获得油酸含量显著提高的转基因大豆新品系。Southern杂交检测表明,外源GmFAD2-1B基因片段已导入大豆基因组,其插入拷贝数为1~5个。qRT-PCR检测表明,外源GmFAD2-1B主要在大豆种子中表达,并导致种子中内源GmFAD2-1 mRNA表达水平显著降低,而根、茎、叶、花组织中内源GmFAD2-1 mRNA表达水平无显著变化。脂肪酸组分分析表明,12份转基因大豆种子油酸含量为27.38%~80.42%,其中,L40和L72油酸含量分别为68.91%~80.42%和65.98%~80.22%,较对照品种Williams 82 (17.8%~22.0%)提高2.65倍以上,亚油酸降低至4.84%~14.55%,饱和脂肪酸降低至10.34%~11.16%,但总脂肪和总蛋白含量与对照品种相比没有显著变化。农艺性状分析表明,转基因大豆在熟期、株高、叶形、花色、结荚高度、百粒重等方面与对照品种也没有显著差异。
| [1]Clemente T E, Cahoon E. Soybean oil: genetic approaches for modification of functionality and total content. Plant Physiol, 2009, 151: 1030–1040 [2]Sari A R, Thomas G, Christoph B, John B O. Contrapuntal networks of gene expression during Arabidopsis seed filling. Plant Cell, 2002, 14: 1191–1206 [3] Okuley J, Lightner J, Feldmann K, Yadav N, Lark E, Browse J. Arabidopsis FAD2 gene encodes the enzyme that is essential for polyunsaturated lipid synthesis. Plant Cell, 1994, 6: 147–158 [4]Li L Y, Wang X L, Gai J Y, Yu D. Molecular cloning and characterization of a novel microsomal oleate desaturase gene from soybean. Plant Physiol, 2007, 64: 516–526 [5] Lee K R, Sohn S I, Jung J H, Kim S H, Roh K H, Kim J B, Suh M C, Kim H U. Functional analysis and tissue-differential expression of four FAD2 genes in amphidiploid Brassica napus derived from Brassica rapa and Brassica oleracea. Gene, 2013, 531: 253–262 [6] Hongtrakul V, Slabaugh M, Knapp S J. A seed specific Δ-12 oleate desaturase is duplicated, rearranged, and weakly expressed in high oleic acid sunflower lines. Crop Sci, 1998, 38: 1245–1249 [7] López Y, Nadaf H L, Smith O D, Fritz A K. Isolation and characterization of the Δ12-fatty acid desaturase in peanut (Arachis hypogaea L.) and search for polymorphisms for the high oleate trait in Spanish market-type lines. Theor Appl Genet, 2000, 101: 1131–1138 [8] Zhang D, Irma L P, Stacy J P, Mongkol N, Purnima N, Sylvia W W, Robert M P, Kent D C. Identification and expression of a new delta-12 fatty acid desaturase (FAD2-4) gene in upland cotton and its functional expression in yeast and Arabidopsis thaliana plants. Plant Physiol Biochem, 2009, 47: 462–471 [9] Heppard E P, Kinney A J, Stecca K L, Miao G H. Developmental and growth temperature regulation of two different microsomalω-6saturase genes in soybeans. Plant Physiol, 1996, 110: 311–319 [10] Tang G Q, Novitzky W P, Griffin H C, Huber S C, Dewey R E. Oleate desaturase enzymes of soybean: evidence of regulation through differential stability and phosphorylation. Plant J, 2005, 44: 433–446 [11] Pham A T, Lee J D, Shannon J G, Bilyeu K D. Mutant alleles of FAD2-1A and FAD2-1B combine to produce soybeans with the high oleic acid seed oil trait. BMC Plant Biol, 2010, 10: 195 [12] Li L Y, Wang X L, Gai J Y, Yu D Y. Isolation and characterization of a seed-specific isoform of microsomal omega-6 fatty acid desaturase gene (FAD2-1B) from soybean. DNA Sequence. 2008. 19: 28–36 [13] Zhang L, Yang X D, Zhang Y Y, Yang J, Qi G X, Guo D Q, Xing G J, Yao Y, Xu W J, Li H Y, Li Q Y, Dong Y S. Changes in oleic acid content of transgenic soybeans by antisense RNA mediated posttranscriptional gene silencing. Int J Genom, 2014, 921–950 [14] Wang G L, Xu Y N. Hypocotyl-based A grobacterium-mediated transformation of soybean (Glycine max) and application for RNA interference. Plant Cell Rep, 2008, 27: 1177–1184 [15]Haun W, Coffman A, Clasen B M, Demorest Z L, Lowy A, Ray E, Retterath A, Stoddard T, Juillerat A, Cedrone F, Mathis L, Voytas D F, Zhang F. Improved soybean oil quality by targeted mutagenesis of the fatty acid desaturase 2 gene family. Plant Biotechnol J, 2014, 12: 934–940 [16] 杨向东, 牛陆, 张伟, 杨静, 杜茜, 邢国杰, 郭东全, 李启云, 董英山. RNAi介导SMV-P3基因沉默增强大豆对花叶病毒病的抗性. 作物学报, 2016, 42: 1647–1655 Yang X D, Niu L, Zhang W, Yang J, Du Q, Xing G J, Guo D Q, Li Q Y, Dong Y S. RNAi-mediated SMV-P3 silencing increases soybean resistance to soybean mosaic virus. Acta Agron Sin, 2016, 42: 1647–1655 (in Chinese with English abstract) [17] Edwards K, Johnstone C, Thompson C. A simple and rapid method for the preparation of plant genomic DNA for PCR analysis. Nucl Acids Res, 1991, 19: 1349 [18]Tel-Zur N, Abbo S, Myslabodski D, Mizrahi Y. Modified CTAB procedure for DNA isolation from epiphytic cacti of the genera Hylocereus and Selenicereus (Cactaceae). Plant Mol Biol Rep, 1999, 17: 249–254 [19] Yoshino M, Nagamatsu A, Tsutsumi K, Kanazawa A. The regulatory function of the upstream sequence of the beta-conglycinin alpha subunit gene in seed-specific transcription is associated with the presence of the RY sequence. Genes Genet Syst, 2006, 81: 135–141 [20] Alt J L, Fehr W R, Welke G A, Shannon J G. Transgressive segregation for oleate content in three soybean populations. Crop Sci, 2005, 45: 2005–2007 [21] Scherder C W, Fehr W R. Agronomic and seed characteristics of soybean lines with increased oleate content. Crop Sci, 2008, 48: 1755–1758 |
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