An Efficient Transformation of Salinity-resistant Peppermint Stem Mediated with Agrobacterium tumefaciens

doi:10.13560/j.cnki.biotech.bull.1985.2016-1199

Abstract

Abstract: Transgenic technology is an important means to improve the essential yield and quality of peppermint（Mentha piperita L.）oil. An Agrobacterium tumefaciens-mediated transformation using bud peppermint stem was established，in order to solve the problems in adventitious bud induction of salinity-resistant peppermint leave，such as easy browning and difficult differentiation. The orthogonal test was carried out to screen the optimal concentration of A. tumefaciens，transformation time and co-culture time. The results showed that the effect of the three factors on transformation displayed as the transformation time（B）> bacterial concentration（A）> co-culture time（C）. The bacterial concentration OD600 = 0.8，the transformation time 15 min and the co-culture time 3 d proved to be the optimal transformation conditions. The optimal composition of transformants was screened to be 1/5MS + 0.5 g/L MES（2-（N-morpholino）ethanesulfonic acid）+ 1% glucose + 2% sucrose and PH 5.4. The transformation was conducted using different plant expression vectors，and the positive plants were detected by genomic PCR and GUS staining. And the results revealed that this transformation system was suitable for the transformation of many plant expression vectors，and the regenerated plants were easily obtained. This provides a new method and system for gene transformation of salinity-resistant peppermint.

Key words: peppermint, stem, Agrobacterium tumefaciens, plant expression vector, genomic PCR, GUS staining

ZHAO Zhong-juan, WEI Yan-li ,LI Ji-shun, WANG Yi-lian ,YANG He-tong. An Efficient Transformation of Salinity-resistant Peppermint Stem Mediated with Agrobacterium tumefaciens[J]. Biotechnology Bulletin, 2017, 33(7): 126-132.

References

[1] Murray MJ, Lincoln DE, Marble PM. Oil composition of Mentha aquatica×M. spicata F1 hybrids in relation to the origin of M.×piperita[J] . Canadian Journal of Genetics and Cytology, 1972, 14：13-29.
[2] Brody AL, Mukhin AG, La Charite J, et al. Up-regulation of nicotinic acetylcholine receptors in menthol cigarette smokers[J] . Int J Neuropsychopharmacol, 2012, 21：1-10.
[3] Pan R, Tian Y, Gao R, et al. Central mechanisms of menthol-induced analgesia[J] . J Pharmacol Exp Ther, 2012, 343：661-672.
[4] Patel R, Gon?alves L, Leveridge M, et al. Anti-hyperalgesic effects of a novel TRPM8 agonist in neuropathic rats：A comparison with topical menthol[J] . Pain, 2014, 155（10）：2097-2107.
[5] Li Z, Yang HT, Wu XQ, et al. Some aspects of salinity responses in peppermint（Mentha piperita L.）to NaCl treatment[J] . Protoplasma, 2015, 252（3）：885-899.
[6] 李哲, 吴晓青, 魏艳丽, 等. NaCl 胁迫对椒样薄荷生长剂抗氧化系统的影响[J] . 广东农业科学, 2014, 41（3）：46-51.
[7] 李哲, 吴晓青, 赵晓燕, 等. NaCl 胁迫对椒样薄荷光合作用和PSII光化学活性的影响[J] . 山东科学, 2014, 27（1）：27-33.
[8] Barton KA, Binns AN, Matzke AJM, et al. Regeneration of intact tobacco plants containing full length copies of genetically engineered T-DNA, and transmission of T-DNA to R1 progeny[J] . Cell, 1983, 32（4）：1033-1043.
[9] 柴志坚, 张芳, 黄园园, 等. 根瘤农杆菌基因工程[J] . 分子植物育种, 2016, 14（1）：92-97.
[10] 许红梅, 张立军, 刘淳. 农杆菌蘸花法侵染拟南芥的研究[J] . 北方园艺, 2010, 14：143-146.
[11] Horsch RB, Fry JE, Hoffmann NL, et al. A simple and general method for transferring genes into plants[J] . Science, 1985, 227：1229-1231.
[12] Girijashankar V, Swathisree V. Genetic transformation of Sorghum bicolor[J] . Physiology and Molecular Biology of Plants, 2009, 15（4）：287-302.
[13] Yasmeen A. An improved protocol for the regeneration and transformation of tomato（cv Rio Grande）[J] . Acta Physiologiae Plantarum, 2009, 31：1271-1277.
[14] 陆玉建, 张韩杰, 韩文瑜, 等. 农杆菌介导rd29A启动子驱动otsB基因转化紫茉莉的研究[J] . 植物研究, 2016, 3：401-408.
[15] Ajenifujah-Solebo SOA, Ingelbrecht I, Isu NR, et al. Reproducible Agrobacterium-mediated transformation of nigerian cultivars of tomato（Solanum lycopersicum L.）[J] . American Journal of Experimental Agriculture, 2014, 4（7）：797-808.
[16] Sujatha M, Sailaja M. Stable genetic transformation of castor（Ricinus communis L.）via Agrobacterium tumefaciens-mediated gene transfer using embryo axes from mature seeds[J] . Plant Cell Reports, 2005, 23（12）：803-810.
[17] Sailaja M, Tarakeswari M, Sujatha M. Stable genetic transformation of castor（Ricinus communis L.）via particle gun-mediated gene transfer using embryo axes from mature seeds[J] . Plant Cell Reports, 2008, 27（9）：1509-1519.
[18] 徐丹丹, 王丕武, 赵邯郸, 等. 农杆菌介导phyA 基因转化玉米愈伤组织培养体系的优化[J] . 湖北农业科学, 2015, 54（2）：465-467.
[19] 方星, 胡倩倩, 王建国, 等. 大豆悬浮细胞培养及作为外源基因转化受体的研究[J] . 大豆科学, 2015, 34（1）：36-41.
[20] Niu X, Hasegawa PM, Bressan RA, et al. Transgenic peppermint（Mentha×piperita L.）plants obtained by cocultivation with Agrobacterium tumefaciens[J] . Plant Cell Reports, 1998, 17（3）：165-171.
[21] Diemer F, Jullien F, Faure O, et al. High efficiency transformation of peppermint（Mentha×piperita L.）with Agrobacterium tumefaciens[J] . Plant Science, 1998, 136（1）：101-108.
[22] Niu X, Li X, Veronese P, et al. Factors affecting Agrobacterium tumefaciens -mediated transformation of peppermint[J] . Plant Cell Reports, 2000, 19（3）：304-310.
[23] Lange BM, Mahmoud SS, Wildung MR, et al. Improving peppermint essential oil yield and composition by metabolic engineering[J] . Proc Natl Acad Sci USA, 2011, 108（41）：16944-16949.
[24] 陈磊, 郭政宏, 程海丽, 等. 农杆菌介导的花魔芋遗传转化体系的优化[J] . 安徽农业科学, 2015, 43（5）：21-24.
[25] Bugbee BG, Salisbury FB. An evaluation of MES（2（N-Morpho-lino）ethanesulfonic acid）and Amberlite IRC-50 as pH buffers for nutrient solution studies[J] . Journal of Plant Nutrition, 1985, 8（7）：567-583 .