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then t ransfer t ransgenes t o f urther g enerations i n t he r ealistic ag ro-ecosystem w here u n- cropped and cropped areas coexist. In this study, we aim to investigate the behavior of the progenies of spontaneous hybrids between w ild B. j uncea and a he rbicide-resistant g ermplasm o f B. nap us similar to th ose commercialized cultivars under the Clearfield® technology. We studied the formation of F2 and BC1 progenies, whether they may grow and reproduce under field conditions, and whether their p erformances ar e ch anged acco rding t o t he p resence o f t he he rbicide-resistance g ene. Instead of exploring the possibility of transgene introgression into the wild species, which has been a lready explored [17], t his w ork f ocused on t he l ikelihood of t he e mergence of backcrossed plants that could behave like B. napus volunteer-like plants. Materials and methods Hybrids A w ild a ccession of Brassica j uncea (2n=36, AABB) from T urkey w as obt ained f rom t he INRA laboratory collection of weeds (ref. X04-021) in Dijon, France. A mutant B. napus cv. Brutor, ( 2n=38, A ACC), b earing a s ingle-gene dom inant r esistance t o c hlorsulfuron, a sulfonylurea he rbicide i nhibiting t he A cetoLactate S ynthase (ALS), w as pr ovided b y M . Renard [ 18]. T hese t wo s pecies were grown i n mixture i n i nsect-proof c ages c ontinuously provided w ith houseflies, Musca dom estica, i n order t o f acilitate r andom pol lination. T he seeds o f B. j uncea were s own i n a g reenhouse at 22°C unde r na tural l ight, a nd t hen t wo- leaved seedlings were sprayed with 2.5 g a.i. ha -1 chlorsulfuron in 300 L of water (Glean, Du Pont de Nemours, France). Eleven out of 8,238 seedlings survived (0.13 %), and these plants were confirmed t o be i nterspecific h ybrids s ince t hey s howed m olecular m arkers of bot h parents (not shown). Progeny of hybrids Branches of all the hybrid plants were cut and planted in the soil to multiply the hybrids. The hybrid cuttings were grown in the greenhouse and entirely covered by a pollen-proof net. Anthers of f lowers o f 2 4 c uttings f rom a ll e leven h ybrids were r emoved a nd t he s tigmas 61
eceived p ollens f rom wild B. j uncea by h and t o g enerate a BC1 generation ( BC1J). O n 2 2 other c uttings, e masculated f lowers r eceived pollens f rom a c onventional he rbicide- susceptible B. napus cv. Drakkar to obtain a BC1 generation (BC1N) segregating for herbicide resistance and s usceptibility (although not yet i dentified a t t hat s tage). The r est o f h ybrid cuttings were kept isolated by a pollen-proof net and served to get a bulked F2 generation (Fig. 2.3). Pollen fertility was checked by staining pollens of 5 f lowers of each of 6 h ybrids with Alexander s olution. N umber of pol linated f lowers, s eed num ber, p er-seed w eight an d germination were recorded. Five hundred F2 seeds, 100 BC1N and 50 BC1J seeds were sown in t he g reenhouse a nd sprayed w ith t he he rbicide G lean a s de scribed above, a nd t hen seedlings alive were counted. Fig. 2.3. Crossing chart indicating how the plant material was obtained and its phenotype for the herbicide-resistance (R or S) 62
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UNIVERSITE DE BOURGOGNE THÈSE Pour
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REMERCIEMENTS My heartfelt thanks g
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forming many of the ideas I present
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Abstract In the framework of commer
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2.3 A rticle 2: Les r étrocroiseme
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Table 3.2. Parameters of linear reg
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experiment. BC1NS is separated into
Page 15 and 16: Fig. 4.7. Mean and standard error (
Page 17 and 18: INTRODUCTION GENERALE Dans le cadre
Page 19 and 20: chapitre 1 dressant l’état des c
Page 21 and 22: CHAPTER 1 LITERATURE REVIEW ON THE
Page 23 and 24: oilseed rape and B. oleracea is ver
Page 25 and 26: populations is reduced as the dista
Page 27 and 28: into hybrids would affect the morph
Page 29 and 30: Snow e t a l. ( 1999) f ound no f i
Page 31 and 32: 1.5 Consequences of introgression I
Page 33 and 34: more flowers and larger plant size
Page 35 and 36: Moreover, beside the introgression
Page 37 and 38: their parental plants. Moon et al.
Page 39 and 40: 2.1 Introduction CHAPTER 2 CONDITIO
Page 41 and 42: ARTICLE 1 The effect of seed size o
Page 43 and 44: plant growth was evaluated without
Page 45 and 46: These two experiments were kept wee
Page 47 and 48: interactions were found among the t
Page 49 and 50: 2006) or m ore f it t han their pa
Page 51 and 52: they b ear b eneficial t ransgenes
Page 53 and 54: Diggle PK, Abrahamson NJ, Baker RL
Page 55 and 56: Ramachandran S , B untin G D, A ll
Page 57 and 58: Table 2.2. F -values f rom a f ive-
Page 59 and 60: Days to flowering Biomass Per. of s
Page 61 and 62: 2.3 Article 2: Les rétrocroisement
Page 63 and 64: Photo.2.3. S praying chlorsulfuron
Page 65: Introduction In t he f ramework o f
Page 69 and 70: Statistical analysis Mean values ar
Page 71 and 72: Characteristics of the backcrosses
Page 73 and 74: Table 2.7. Mean (±95% CL) of plant
Page 75 and 76: plants w ith B. napus morphology t
Page 77 and 78: Productivity of the resistant proge
Page 79 and 80: References [1] A .A. Snow, D .A. A
Page 81 and 82: insight into adaptive life-history
Page 83 and 84: CHAPTER 3 EFFETS DE LA RESISTANCE A
Page 85 and 86: 3.2 A rticle 3 : S imulation d e l
Page 87 and 88: compétition et le devenir des popu
Page 89 and 90: difficult to predict. Similarly, it
Page 91 and 92: Table 3.1: ANOVA results of the eff
Page 93 and 94: Table 3.2: Parameters of linear reg
Page 95 and 96: herbivores than when exposed to Bt
Page 97 and 98: esistant hybrid populations might i
Page 99 and 100: Letourneau D .K., H agen J .A., 200
Page 101 and 102: Germination rate 0.6 0.5 0.4 0.3 0.
Page 103 and 104: 3.3 Article 4: Compétition entre p
Page 105 and 106: Introduction The invasion of transg
Page 107 and 108: Whether a popul ation w ill e volve
Page 109 and 110: while ensuring that the same ratio
Page 111 and 112: silique number, seed number, seed w
Page 113 and 114: Our ga rden e xperiments s howed t
Page 115 and 116: experiment, harsh growth conditions
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References Agrawal AA. 2004. R esis
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density- dependent c osts of vi ral
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Table 3.5. F values of the analysis
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Table 3.7. Results of Helmert contr
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Fig. 3.4. Examples of experimental
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Flowering date Seed weight Biomass
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No. viable seeds Biomass 50000 4000
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Photo 3.3. Cotton bollworm (Helicov
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Introduction Spontaneous introgress
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screening as transgenic BC2 (trBC2)
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2002), but certain studies showed n
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Acknowledgements We t hank Z hixi T
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92, 368-374. Rieseberg, LH, and Bur
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Table 3.10. F-values of three-way A
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Seed number Biomass 140 120 100 80
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CHAPTER 4 RECHERCHE DES CONSEQUENCE
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Photo 4.1. Wild radish in a herbici
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Introduction Plant divergence is ge
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or t he popul ation of wild r adish
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estimated. All the seeds of every h
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Field experiment Number of siliques
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aphanistrum from hybrids and R. sat
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competition diminished the differen
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Conner, J. K., Rice A. M., Stewart
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Rattenbury J A. 1962. C yclic h ybr
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Fig. 4.1. Four r egions where w ild
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Fig. 4.3. Flower phot os showing pe
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Fig. 4.5. Plot of the first two axe
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Plant weight No. of seeds per plant
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Emergence rate Plant weight No.of a
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CONCLUSION 172
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véritable s uccès ad aptatif d an
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Dans l e cas d es p remières gén
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REFERENCES Abbott RJ, Comes HP.2007
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Bing D J, D owney R K and R akow G
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carinata and Brassica rapa. Plant B
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Hybridisation within Brassica and a
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etween weedy Brassica rapa and oils
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Hybridization between oilseed rape
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Nosil P. 2008. Speciation with gene
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32:305-32 Schadler, M., R. Brandl,
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Thalmann C, Guadagnuolo R, Felber F
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Zheng XM and Ge S. 2010. Ecological
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Therefore, after a review of the st
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more easily sieved out by harvester
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Transgenic F1 produced higher bioma
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