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IUFRO Working Party 2.09.02: Somatic Embryogenesis and Other Vegetative Propagation Technologies, Vitoria 2014 www.bosquesnaturales.com Biotechnological.

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Presentation on theme: "IUFRO Working Party 2.09.02: Somatic Embryogenesis and Other Vegetative Propagation Technologies, Vitoria 2014 www.bosquesnaturales.com Biotechnological."— Presentation transcript:

1 IUFRO Working Party 2.09.02: Somatic Embryogenesis and Other Vegetative Propagation Technologies, Vitoria 2014 www.bosquesnaturales.com Biotechnological Approaches for Walnut Wood Production Ricardo Julián Licea Moreno 1,2, Angela Contreras 2, Irene Merino 2, Julia Quintana 2, Ignacio Urban 3, Marcos Daquinta Gradaille 4, Luis Gomez 2 1 Department of Biotechnology, Micropropagation Unit, Bosques Naturales S. A. Ave. de la Vega, 1, 28108 Madrid, Spain ricardolicea@bosquesnaturales.comricardolicea@bosquesnaturales.com 2 Department of Biotechnology, Center for Plant Biotechnology and Genomics, Universidad Politécnica de Madrid, Spain 3 Department of Forestry, Bosques Naturales S. A., Spain 4 Bioplantas Center, Universidad de Ciego de Avila, Cuba

2 IUFRO Working Party 2.09.02: Somatic Embryogenesis and Other Vegetative Propagation Technologies, Vitoria 2014 www.bosquesnaturales.com Outline 1.Basic lines of BN’s selection program for high quality wood production 2.Micropropagation of èlite trees: the key factors and the last results 3.Adaptability trials for micropropagated walnut clones 4.Differential expression profiles of walnut chaperones

3 IUFRO Working Party 2.09.02: Somatic Embryogenesis and Other Vegetative Propagation Technologies, Vitoria 2014 www.bosquesnaturales.com Walnut hybrid ( Juglans major var. 209 (Torrey) Heller x J. regia L.) -Clime adaptation - Vigour - Apical dominance - Straighness - Late sprouting 12 years-old trees

4 IUFRO Working Party 2.09.02: Somatic Embryogenesis and Other Vegetative Propagation Technologies, Vitoria 2014 www.bosquesnaturales.com Forest Management Unit Site Characteristics GeronaCáceresNavarra Coordinates 42º 09' 55'' N 02º 56' 37'' E 40º 06' 54'' N 05º 20' 32'' W 42º 10' 10'' N 01º 39' 09'' W Altitude (m)450 m350 m250 m Mean Annual Temperature (ºC)14.714.113.8 Total Annual Precipitation (mm)857844416 Type of ManagementIntensive (Fertirrigation) Extensive Age at Sampling (years)171816 Sib Progeny (trees)22105332835007 GeronaCáceresNavarra

5 IUFRO Working Party 2.09.02: Somatic Embryogenesis and Other Vegetative Propagation Technologies, Vitoria 2014 www.bosquesnaturales.com 15 years old trees

6 IUFRO Working Party 2.09.02: Somatic Embryogenesis and Other Vegetative Propagation Technologies, Vitoria 2014 www.bosquesnaturales.com 10 years old clonal plantation

7 IUFRO Working Party 2.09.02: Somatic Embryogenesis and Other Vegetative Propagation Technologies, Vitoria 2014 www.bosquesnaturales.com FeEDTAFeEDDHA + Phloroglucinol Moreno et al. 2012

8 IUFRO Working Party 2.09.02: Somatic Embryogenesis and Other Vegetative Propagation Technologies, Vitoria 2014 www.bosquesnaturales.com * Without repetition Genotype PG (mg/L) ControlMycorrhizeMycorrhize+Bacteria Rooted Microshoots Survivor Microshoots Survival (%) Rooted Microshoots Survivor Microshoots Survival (%) Rooted Microshoots Survivor Microshoots Survival (%) D53 0803442,59222,2*402050,0* 251608653,81117264,9946771,3 50491632,7*1206150,81064946,2 Total28913647,124013556,324013656,7 DM50281553,6*282071,4*282175,0* Effect of mycorrhization on vitroplant survival

9 IUFRO Working Party 2.09.02: Somatic Embryogenesis and Other Vegetative Propagation Technologies, Vitoria 2014 www.bosquesnaturales.com Arbuscules Root of control plants Root of mycorrhized plants

10 IUFRO Working Party 2.09.02: Somatic Embryogenesis and Other Vegetative Propagation Technologies, Vitoria 2014 www.bosquesnaturales.com La Mota (Arzúa) Plantation: May, 2013 Latitude: 43° 00´ 00´´ N Longitude: 08° 08´ 00´´ W Altitude: 450 m Clones: 9 July, 2014 June, 2014 El Soto (Toledo) Plantation: May,2014 Latitude: 39° 54´ 05´´ N Longitude: 04° 17´ 54´´ W Altitude: 475 m Clones: 8 August, 2014 IFAPA (Granada) Plantation: March, 2014 Latitude: 37° 10´ 30 ´´ N Longitude: 03° 38´ 10 ´´ W Altitude: 630 m Clones: 8 9 different clones Latinice experimental design 4 replications 4 blocks/ replication 4 trees/plot

11 IUFRO Working Party 2.09.02: Somatic Embryogenesis and Other Vegetative Propagation Technologies, Vitoria 2014 www.bosquesnaturales.com Sprouting & mortality (Granada clonal trial)

12 IUFRO Working Party 2.09.02: Somatic Embryogenesis and Other Vegetative Propagation Technologies, Vitoria 2014 www.bosquesnaturales.com Chaperones sHSP HSP40 Stress response Hybrid poplar thermoprotective (Merino et al., 2013 & 2014) 3 major walnut’s chaperones JuHSP 17.8 JuHSP 17.9 JuHSP 18.1 D15 & D117 different Western blot profiles unpublished data

13 IUFRO Working Party 2.09.02: Somatic Embryogenesis and Other Vegetative Propagation Technologies, Vitoria 2014 www.bosquesnaturales.com 4°C T0 Control T1 (1 day) T3 (3 days) T5 (5 days) T7 (7 days) RNA Extraction 6 Genotypes cDNA QRT-PCR

14 IUFRO Working Party 2.09.02: Somatic Embryogenesis and Other Vegetative Propagation Technologies, Vitoria 2014 www.bosquesnaturales.com JuHSP17.8 Expression Under Low Temperature

15 IUFRO Working Party 2.09.02: Somatic Embryogenesis and Other Vegetative Propagation Technologies, Vitoria 2014 www.bosquesnaturales.com

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