GENE FLOW FROM TRANSGENIC GLUFOSINATE- OR GLYPHOSATE-TOLERANT OILSEED RAPE TO WILD RAPE

doi:10.17521/cjpe.2007.0093

Abstract

Abstract:

Aims Wild rape (Brassica juncea var. gracilis), a wild relative of oilseed rape (B. napus), was chosen for a case study because gene flow from herbicide-resistant oilseed rape to wild rape rarely has been explored. We asked a) are transgenic herbicide-resistant oilseed rape and wild rape sexually compatible, b) is the fitness of their F₁ improved and c) is gene flow possible?
Methods We used hand pollination to test sexual compatibility and examined resistance of the F₁ by applying herbicide of 2 000 μg a.i.·kg^-1 with 0.1 L·m^-2 volume at 3-4 leaf stage twice and by PCR and examining vegetative growth, pollen viability and seed setting rate. Crossing from herbicide-resistant oilseed rape to wild rape also was tested under open pollination.
Important findings Indexes of sexual compatibility of wild rape×glufosinate-tolerant oilseed and wild rape×glyphosate-tolerant oilseed were 13.87 and 13.83, respectively, under hand pollination, and these indexes were not significantly different from that of the female parent under self pollination and open pollination. All F₁ were significantly resistant to the corresponding herbicide. Moreover, herbicide-resistant gene of F₁ was confirmed by PCR and F₁ of wild rape×glufosinate-resistant oilseed and wild rape×glyphosate-resistant oilseed produced 426 bp and 931 bp identical special band, respectively. The fitness of the F₁ changed compared with that of the female parent. The germination rate of the F₁ was similar with that of the female parent, >98%. Due to taller individuals and larger leaves, the vegetative growth of the F₁ was much better than that of the female parent. However, pollen viabilities of both kinds of F₁ were only half that of the female parent. Furthermore, the seed number per silique of the F₁ was very low, only 0.59 and 0.58 seeds per silique, respectively. The cross rate of wild rape×glufosinate-resistant and wild rape×glyphosate-resistant was 0.02% and 0.01%, respectively, under open pollination. Our study suggests the possibility of gene flow from transgenic rape to wild rape. The possibilities of gene flow from the two herbicide-resistant transgenic oilseed rapes to wild rape were similar under hand pollination or open pollination. However, the effect of gene flow from transgenic herbicide-resistant oilseed to wild rape needs further research.

Key words: transgenic glufosinate-resistant oilseed rape, transgenic glyphosate-resistant oilseed rape, gene flow, fitness

SONG Xiao-Ling, HUANGFU Chao-He, QIANG Sheng. GENE FLOW FROM TRANSGENIC GLUFOSINATE- OR GLYPHOSATE-TOLERANT OILSEED RAPE TO WILD RAPE[J]. Chin J Plant Ecol, 2007, 31(4): 729-737.

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Figures/Tables 9

References 41

[1]	Baranger A, Chèvre AM, Eber F, Renard M (1995). Effect of oilseed rape genotype on the spontaneous hybridization rate with a weedy species :an assessment of transgene dispersal. Theoretical and Applied Genetics, 91, 956-963. DOI URL PMID
[2]	Bing DJ, Downey RK, Rakow GFW(1996). Hybridizations among Brassica napus, B. rapa and B. juncea and their two weedy relatives B. nigra and Sinapis arvensis under open pollination conditions in the field. Plant Breeding, 115, 470-473. DOI URL
[3]	Chen LH(陈梁鸿), Wang XW(王新望), Zhang WJ(张文俊), Zhang XD(张晓东), Hu DF(胡道芬), Liu GT(刘广田)(1999). Transformation of common wheat (Triticum aestivum L.) with herbicide-resistant EPSPs gene. Acta Genetica Sinica (遗传学报), 26, 239-243. (in Chinese with English abstract)
[4]	Chèvre AM, Eber F, Baranger A, Renard M (1997). Gene flow from transgenic crops. Nature, 389, 924. DOI URL
[5]	Chèvre AM, Eber F, Darmency H, Renard M (1999). Last results concerning gene flow from transgenic oilseed rape to wild radish. Proceedings of the 10th International Rapeseed Congress, Canberra, Australia, 5.
[6]	Crawley MJ, Brown SL, Hails RS, Kohn DD, Rees M (2001). Transgenic crops in natural habitats. Nature, 409, 682-683. DOI URL PMID
[7]	Darmency H, Fleury A(2000). Mating system in Hirschfeldia incana and hybridization to oilseed rape. Weed Research, 40, 231-238. DOI URL
[8]	Geng YP(耿宇鹏), Zhang WJ(张文驹), Li B(李博), Chen JK(陈家宽)(2004). Phenotypic plasticity and invasiveness of alien plants. Biodiversity Science (生物多样性), 12, 447-455. (in Chinese with English abstract)
[9]	Goy PA, Duesing JH (1996). Assessing the environmental impact of gene transfer to wild relatives. Biotechnology, 14, 39-40. DOI URL PMID
[10]	Guan CY(官春云)(2005). Breeding of rapeseed by genetic trans-formation. Chinese Journal of Oil Crop Sciences (中国油料作物学报), 27(1), 97-103. (in Chinese with English abstract)
[11]	Guan CY(官春云), Li X(李木旬)(1997). Research and application of transgenic oilseed rape. Chinese Journal of Cell Biology (细胞生物学杂志), 19(1), 18-23. (in Chinese)
[12]	Gueritaine G, Sester M, Eber F, Chèvre AM, Darmency H(2002). Fitness of backcross six of hybrids between transgenic oilseed rape (Brassica napus) and wild radish (Raphanus raphamistrum). Molecular Ecology, 11, 1419-1426. DOI URL PMID
[13]	Guo QY(郭青云), Tu HL(涂鹤龄), Qiu XL(邱学林), Xin CY(辛存岳)(1998). The occurrence dynamics and control technique of wild rape (Brassica juncea (L.) Czern.et Coss.). Science and Technology of Qinhai Agriculture and Forestry (青海农林科技), (4), 38-41. (in Chinese)
[14]	Hauser TP, Jørgensen RB, Østergård H (1998). Fitness of backcross and F₂ hybrids between weedy Brassica rapa and oilseed rape (B. napus). Heredity, 81, 436-443. DOI URL
[15]	Hauser TP, Damgaard C, Jørgensen RB (2003). Frequency-dependent fitness of hybrids between oilseed rape (Brassica napus) and weedy B. rapa (Brassicaceae). American Journal of Botany, 90, 571-578. DOI URL PMID
[16]	Jørgensen RB, Andersen B, Hauser TP, Landbo L, Mikkelsen TR, Østerg⁈rd H (1998). Introgression of crop genes from oilseed rape ( Brassica napus) to related wild species - an avenue for the escape of engineered genes. Acta Horticulturae, 459, 211-217.
[17]	Jørgensen RB, Anderson B, landbo L, Mikkelsen TR (1996). Spontaneous hybridization between oilseed rape ( Brassica napus) and weedy relatives. Acta Horticulturae, 407, 193-200.
[18]	Kerlan MC, Chèvre AM, Eber F, Baranger A, Renard M (1992). Risk assessment of outcrossing of transgenic rapeseed to related species. I. Interspecific hybrid production under optimal condition with emphasis on pollination and fertilization. Euphytica, 62, 145-153. DOI URL
[19]	Kerlan MC, Chèvre AM, Eber F (1993). Interspecific hybrids between a transgenic rapeseed ( Brassica napus) and related species: cytogenetical characterization and detection of the transgene. Genome, 36, 1099-1106. DOI URL PMID
[20]	Kling J (1996). Could transgenic super crops one day breed super weeds? Science, 274, 180-181. DOI URL
[21]	Lefol E, Danielou V, Darmency H(1996). Predicting hybridization between transgenic oilseed rape and wild mustard. Field Crops Research, 45, 153-161 DOI URL
[22]	Lefol E, Seguin-Swartz G, Downey RK (1997). Sexual hybridisation in crosses of cultivated Brassica species with the crucifers Erucatrum gallicum and Raphanus raphanistrum: potential for gene introgression. Euphytica, 95, 127-139. DOI URL
[23]	Lu CM (卢长明), Xiao L (肖玲), Wu YH (武玉花) (2005). Ecological risk assessment of transgenic rapeseed in China. Journal of Agricultural Biotechnology (农业生物技术学报), 13, 267-275. (in Chinese with English abstract)
[24]	Metz PLJ, Jacobsen E, Nap JP, Pereira A, Stiekema WJ (1997). The impact on biosafety of the phosphinothricin-tolerance transgene in inter-specific B. rapa×B. napus hybrids and their successive backcrosses. Theoretical and Applied Genetics, 95, 442-450. DOI URL
[25]	Pan LW (潘良文), Chen JH (陈家华), Shen YF (沈禹飞), Hu YQ (胡永强), Tao J (陶军), Han W (韩伟) (2001). Detection comparison of CP4-EPSPS gene from genetically modified Roundup-Ready rapeseed and soyabean. Letters in Biothechnology (生物技术通讯), 12, 175-177, 207. (in Chinese with English abstract)
[26]	Pu HM (蒲惠明), Qi CK (戚存扣), Fu SZ (傅寿仲) (1991). A new method for estimating pollen viability of rapeseed-exosmosis of metabolic products. Chinese Journal of Oil Crop Sciences (中国油料作物学报), (2), 79-81. (in Chinese with English abstract)
[27]	Pu HM(浦惠明), Zhang JF(张洁夫), Qi CK(戚存扣), Gao JQ(高建芹), Chen XJ(陈新军) (2003). Inheritance of herbicide resistance in Brassica napus and its utilization. Jiangsu Journal of Agricultural Sciences (江苏农业科学), 19(2), 81-86. (in Chinese with English abstract)
[28]	Pu HM(浦惠明), Qi CK(戚存扣), Zhang JF(张洁夫), Fu SZ(傅寿仲), Gao JQ(高建芹), Chen XJ(陈新军), Chen S(陈松), Zhao XX(赵祥祥) (2005). The studies on gene flow from GM herbicide-tolerant rapeseed to cruciferous weeds. Acta Ecologica Sinica (生态学报), 25, 910-916. (in Chinese with English abstract)
[29]	Qian YQ (钱迎倩), Wei W (魏伟), Tian Y (田彦), Ma KP (马克平) (1999). Application and potential problems of transgenic crops. Chinese Journal of Applied and Environmental Biology (应用与环境生物学报), 5, 427-433. (in Chinese with English abstract)
[30]	Qiang S (强胜) (2001). The opportunities and challenges of weed science facing to the bioscience era. World Agriculture (世界农业), 264(4), 37-39. (in Chinese)
[31]	Rieger MA, Potter TD, Preston C, Powles SB(2001), Hybridisation between Brassica napus L. and Raphanus raphanistrum L. under agronomic field conditions. Theoretical and Applied Genetics, 103, 555-560. DOI URL
[32]	Schefler JA, Dale PJ (1994). Opportunities for gene transfer from transgenic oilseed rape ( Brassica napus) to related species. Transgenic Research, 3, 263-278. DOI URL
[33]	Snow AA, Palma PM (1997). Commercialization of transgenic plants: potential ecological risks. BioScience, 47, 86-96. DOI URL
[34]	Song XL (宋小玲), Qiang S (强胜) (2003). Sexual compatibility of three species of oilseed rapes (Brassica spp.) with wild rapes (B. juncea var. gracilis Tsenet Lee) and the fitness of F₁-potential for gene transfer. Chines Journal of Applied and Environmental Biology (应用与环境生物学报), 9, 357-361. (in Chinese with English abstract)
[35]	Song ZP, Lu BR, Wang B, Chen JK (2004). Fitness estimation through permance comparison of F₁ hybrids with their parental species Oryza rufipogon and O. sativa. Annals of Botany, 93, 311-316. DOI URL PMID
[36]	Su SQ (苏少泉) (2005). Glufosinate review. Chinese Journal of Pesticides (农药), 44, 529-532. (in Chinese with English abstract)
[37]	Vacher C, Weis AE, Hermann D, Kossler T, Young C, Hochberg ME (2004). Impact of ecological factors on the initial invasion of Bt transgenes into wild populations of birdseed rape (Brassica rapa). Theoretical and Applied Genetics, 109, 806-814. DOI URL
[38]	Wang XF (王新发), Wang HZ (王汉中), Liu GH (刘贵华), Hu ZM (胡赞民), Zheng YB (郑元本)(2005). Transgenic hybrid parents in Brassica napus transformed with bivalent genes for resistance to Sclerotinia sclerotiorum. Chinese Bulletin of Botany (植物学通报), 22, 292-301. (in Chinese with English abstract)
[39]	Warwick SI, Simard MJ, Legere A, Beckie HJ, braun L, Zhu B, Mason P, Segain-Swartz G, Stewart CN Jr (2003). Hybridization between transgenic Brassica napus L. and its wild relatives: Brassica rapa L.,Raphanus raphanistrum L., Sinapis arvensis L., and Erucastrum gallicum (Willd.) O. E. Schulz. Theoretical and Applied Genetics, 107, 528-539. DOI URL PMID
[40]	Wei W (魏伟), Qian YQ (钱迎倩), Ma KP (马克平)(1999). Gene flow between transgenic crops and their wild related species. Acta Botanica Sinica (植物学报), 41, 343-348. (in Chinese with English abstract)
[41]	Yang FY (杨逢玉), Zhang HJ (张宏军), Ni HW (倪汉文) (2002). Mechanism and application of non-selective herbicide gluphosinate. Journal of Beijing Agricultural College (北京农学院学报), 17(4), 100-105. (in Chinese)

被测基因 Detected gene	引物序列 Primer sequence	预期扩增产物长度 Expected product length (bp)
EPSPS	P1:5'-GGT-CCT-TCA-TGT-TCG-GCG-GTC-3'	931
	P2:5'-CTC-GCC-CTC-ATC-GCA-ATC-CAC-3'
bar	P1:5'-CGA-GAC-AAG-CAC-GGT-CAA-CTT-3'	426
	P2:5'-AAA-CCC-ACG-TCA-TGC-CAG-TTC-3'

被测基因 Detected gene	引物序列 Primer sequence	预期扩增产物长度 Expected product length (bp)
EPSPS	P1:5'-GGT-CCT-TCA-TGT-TCG-GCG-GTC-3'	931
	P2:5'-CTC-GCC-CTC-ATC-GCA-ATC-CAC-3'
bar	P1:5'-CGA-GAC-AAG-CAC-GGT-CAA-CTT-3'	426
	P2:5'-AAA-CCC-ACG-TCA-TGC-CAG-TTC-3'

杂交组合 Combination	授粉花蕾数(个) NFP	收获种子数(粒) NSH	亲和性指数 IC	角果长度 LS (cm)
野芥菜自交 Wild rape self-pollination	201	2 623	13.05^Aa	2.80±0.31^Aa
野芥菜开放授粉 Wild rape open-pollination	280	3 850	13.75^Aa	2.82±0.28^Aa
与抗草丁膦油菜杂交 Wild rape×glufosinate-resistant oilseed rape	511	7 087	13.87^Aa	2.79±0.24^Aa
与抗草甘膦油菜杂交 Wild rape×glyphosate-resistant oilseed rape	502	6 943	13.83^Aa	2.71±0.27^Aa

杂交组合 Combination	授粉花蕾数(个) NFP	收获种子数(粒) NSH	亲和性指数 IC	角果长度 LS (cm)
野芥菜自交 Wild rape self-pollination	201	2 623	13.05^Aa	2.80±0.31^Aa
野芥菜开放授粉 Wild rape open-pollination	280	3 850	13.75^Aa	2.82±0.28^Aa
与抗草丁膦油菜杂交 Wild rape×glufosinate-resistant oilseed rape	511	7 087	13.87^Aa	2.79±0.24^Aa
与抗草甘膦油菜杂交 Wild rape×glyphosate-resistant oilseed rape	502	6 943	13.83^Aa	2.71±0.27^Aa

母本 Female	萌发率 Germination rate(%)
母本 Female	自交 Self-pollination	× 抗草丁膦油菜 ×glufosinate-resistant	× 抗草甘膦油菜 × glyphosate-resistant
野芥菜 Wild rape	100.00^Aa	98.34^Aa	98.89^Aa