Experimental tobacco plot size and design to breeding lines evaluation
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1 Experimental tobacco plot size and design to breeding lines evaluation CORESTA CONGRESS September 2012 Sapporo, Japan
2 Experimental tobacco plot size and design to breeding lines evaluation TOLEDO, F.H.R.B.(1); RAMALHO, M.A.P.(2); PULCINELLI, C.E.(3); BRUZI, A.T.(4) (1) Department of Genetics, University of São Paulo, Piracicaba, Brazil; (2) Biology Department, Federal University of Lavras, Minas Gerais, Brazil; (3) Souza Cruz S.A., Tobacco Breeding Centre, Rio Negro, Brazil; (4) Agriculture Department, Federal University of Lavras, Minas Gerais, Brazil
3 Introduction Value for Cultivation and Use: VCU The experimental Value for Cultivation and Use (Piepho and Mohring, 2006) is a requirement of the Ministry of Agriculture, Livestock and Supply (MAPA) (Law Number 9456 of ), for the new cultivars release (BRAZIL, 1997); The rules for the VCU carry out are different according to the species and are set by the MAPA; Several aspects should be considered in the experimental strategy of VCU's, including the size, plot design and number of replications;
4 Introduction Value for Cultivation and Use:VCU To the tobacco crop information regarding to the plot size and design are scarce (FEHR, 1987). In the case of tropical and subtropical conditions were not found references. The most usual plot size for tobacco lines evaluation is formed by six rows, with seven plants per line (CREW and JOHNES, 1972; GUPTON and ARCHER, 1968); This relatively large plot limits, the number of the new lines to be tested (close to 10 lines).
5 Introduction Value for Cultivation and Use:VCU For the tobacco crop the rules for the VCU carry out were not defined; And tow major questions should be answered;
6 Introduction Value for Cultivation and Use:VCU Is it necessary the use of plots with borders? How is the number of plants per plot and their design to obtain reliable estimates of the cultivars mean under evaluation?
7 Objective The aim with this work was to obtain information about the strategies for carrying out Tobacco VCU trials to be evaluated at southern Brazil, in relation to the plot size and design.
8 Material and Methods Local The data was recorded from two different trials (for the set of 10 Burley and Virginia breeding lines), carried out by the Souza Cruz Company, in Rio Negro, Paraná State, Southern Brazil by 2009/2010 season.
9 Material and Methods Local The data was recorded from two different trials (for the set of 10 Burley and Virginia breeding lines), carried out by the Souza Cruz Company, in Rio Negro, Paraná State, Southern Brazil by 2009/2010 season.
10 Material and Methods Experimental Design and Evaluated Traits The experimental design was randomized complete blocks design with four replications, with ten lines in the final stage of evaluation. The plot was performed by six rows of seven plants each, total of 42 plants per plot. From each plant, referencing the position of its plant in the plot (row and column), was recorded the following traits: cured leaf weight (g/plant), total sugar content (%) and total alkaloids content (%);
11 Material and Methods Plot Arrangement: Row and Column 7 Utile Area Columns Rows Data was recorded from individual plants Border Area
12 Material and Methods Experimental Plan: Trial Map 37,0 meters Front Line Sense Lenght Back 1 Plot CMF Line 1 Treat 6 lines of 7 plants 7.2 m x 3.5 m 36,00 meters Left Front of the Trial Right Corridor 1 m Plot Line CMF CMF CMF CMF CMF Treat Corridor 1 m Replication I CMF CMF CMF CMF CSC Corridor 1 m CMF CMF CMF CSC 444 CMF Corridor 1 m CMF CMF CMF CMF CMF Replication II Corridor 1 m CMF CMF CMF CMF CMF Corridor 1 m CSC 444 CMF CMF CMF CMF Replication III Corridor 1 m CSC 444 CMF CMF CMF CMF Corridor 1 m CMF CMF CMF CMF CMF Replication IV Corridor 1 m Back of the Trial
13 Material and Methods Data Analysis and Estatistical Procedure It was performed the variance analysis following the split- plot model according to Pimentel-Gomes (2009) in order to answer the question: Is it necessary plots with border?
14 Material and Methods Data Analysis and Estatistical Procedure The variance analysis was performed using three different schemes: Between location in the plot (Utile or Border plants); Between rows; Between columns; Columns 7 Utile Area Border Area Rows
15 Material and Methods Data Analysis and Estatistical Procedure Estatistical Model in which: Y ijk = μ+r i +t j +(tr) ij +s k +(rs) ik +(ts) jk +e ijk y ijk is the observation in the replication i, treatment j, position k; μ is a constant, the average of observation; r i is the replication i, with i = 1, 2,..., 4; t j is treatment j, with j = 1, 2,..., 10; (tr) ij is the error (a), between plots; s k is the position effect; (rs) ik is the error (b); (ts) ik is the interaction effect treatment i position k; and ε ijk experimental error, error (c), with ε N(0, σ 2 ).
16 Material and Methods Data Analysis and Estatistical Procedure It was also performed the resampling method (simulation) and Maximum Curvature Method of the Variation Coefficient (CV%) in order to answer the question: How is the number of plants per plot and their design to obtain reliable estimates of the cultivars mean under evaluation?
17 Material and Methods Data Analysis and Estatistical Procedure The resampling method was utilized as follows: an algorithm (bootstrap) on the data sheet draw lots of n plants for each plot, and the variance analysis was carried out for each one of these lots; The number of sampled plants (n) varied from 2 to 41 randomly selected without replacement and the process was repeated 2000 times for each n; All analyze were performed and/or implemented using the software R (R Development Core Team, 2011). Columns 7 Utile Area Rows Border Area
18 Material and Methods Data Analysis and Estatistical Procedure For the maximum curvature of the coefficient of variation estimation the variance analysis errors was used to estimate the autocorrelation coefficient of first-order (ρ); According to Parnaiba, Francis & Mitchell (2009) the point of maximum curvature of the coefficient of variation can be obtained by;
19 Material and Methods Data Analysis and Estatistical Procedure Maximum curvature of the coefficient of variation in which: X 0 is the maximum curvature point of coefficient variation (CV%); ρ is the correlation coefficient; σ 2 is the experimental error variance; and Z is the plot average.
20 Results and Discussion Table 1. Summary of variance analysis for Burley varietal tobacco group, for cured leaf weight (g / plant), sugar content (%) and total alkaloids content (%), considering the border and the utile area plot. Rio Negro, Parana, 2009/2010. Source of Variation D.F. Curead Leaves (g/plant) Total Sugar (%) Tota Alkalloids (%) M.S. P (value) M.S. P (value) M.S. P (value) Replication < Lines (L) < 0.01 Error (a) Border vs Utile (P) < < 0.01 Error (b) Interaction L x P Error Average Accuracy Border Utile
21 Results and Discussion Table 2. Summary of variance analysis for Virginia varietal tobacco group, for cured leaf weight (g / plant), sugar content (%) and total alkaloids content (%), considering the border and the utile area plot. Rio Negro, Parana, 2009/2010. Source of Variation D.F. Curead Leaves (g/plant) Total Sugar (%) Tota Alkalloids (%) M.S. P (value) M.S. P (value) M.S. P (value) Replication < < 0.01 Lines (L) < < 0.01 Error (a) Border vs Utile (P) < < 0.01 Error (b) Interaction L x P Error Average Border Utile Accuracy
22 Results and Discussion The results for the two tobacco varieties groups for the three traits were very similar. The interaction lines position was not significant for the three traits. Buy another way, the behavior of the lines was similar regardless they were in the border or useful area.
23 Results and Discussion Table 5. Summary of variance analysis for Burley varietal tobacco group, for cured leaf weight (g/plant), sugar content (%) and total alkaloids content (%), considering the different plot rows. Rio Negro, Parana, 2009/2010. Source of Variation D.F. Curead Leaves (g/plant) Total Sugar (%) Tota Alkalloids (%) M.S. P (value) M.S. P (value) M.S. P (value) Replication ,55 0,08 10,11 < 0,01 2,67 0,05 Lines (L) ,19 0,06 1,08 0,57 3,49 < 0,01 Error (a) ,26 1,26 0,92 Betweenn Rows (P) ,44 0,42 0,18 0,03 0,29 0,06 Error (b) ,20 0,11 0,17 Interaction L x P ,46 0,86 0,05 0,86 0,20 0,04 Error ,44 0,07 0,13
24 Results and Discussion Table 6. Summary of variance analysis for Virginia varietal tobacco group, for cured leaf weight (g/plant), sugar content (%) and total alkaloids content (%), considering the different plot rows. Rio Negro, Parana, 2009/2010. Source of Variation D.F. Curead Leaves (g/plant) Total Sugar (%) Tota Alkalloids (%) M.S. P (value) M.S. P (value) M.S. P (value) Replication ,01 < 0,01 7,38 0,55 0,68 < 0,01 Lines (L) ,71 < 0,01 5,55 0,83 0,68 < 0,01 Error (a) ,96 10,25 0,11 Betweenn Rows (P) 5 570,02 0,22 3,65 0,01 0,05 0,07 Error (b) ,77 1,17 0,02 Interaction L x P ,34 0,36 1,02 0,59 0,03 0,26 Error ,61 1,09 0,03
25 Results and Discussion Table 7. Summary of variance analysis, for Burley varietal group, for cured leaf weight (g/plant), sugar content (%) and total alkaloids content (%), considering the different columns of the plot. Rio Negro, Parana, 2009/2010. Source of Variation D.F. Curead Leaves (g/plant) Total Sugar (%) Tota Alkalloids (%) M.S. P (value) M.S. P (value) M.S. P (value) Replication ,27 0,08 11,80 < 0,01 3,12 0,05 Lines (L) ,87 0,06 1,26 0,57 4,08 < 0,01 Error (a) ,23 1,47 1,08 Betweenn Columns (P) ,13 < 0,01 0,03 0,35 0,61 < 0,01 Error (b) ,62 0,02 0,11 Interaction L x P ,43 0,10 0,03 0,26 0,12 0,03 Error ,72 0,02 0,08
26 Results and Discussion Table 8. Summary of variance analysis, for Virginia varietal group, for cured leaf weight (g/plant), sugar content (%) and total alkaloids content (%), considering the different columns of the plot. Rio Negro, Parana, 2009/2010. Source of Variation D.F. Curead Leaves (g/plant) Total Sugar (%) Tota Alkalloids (%) M.S. P (value) M.S. P (value) M.S. P (value) Replication ,42 < 0,01 8,57 0,55 0,80 < 0,01 Lines (L) ,63 < 0,01 6,53 0,83 0,78 < 0,01 Error (a) ,20 12,02 0,12 Betweenn Columns (P) ,77 < 0,01 1,48 0,07 0,20 < 0,01 Error (b) ,63 2,28 0,04 Interaction L x P ,71 0,91 0,98 0,09 0,02 0,23 Error ,04 0,74 0,02
27 Results and Discussion It was found that there were no differences in average performance of lines in the different rows in the plot; The average performance of the lines varied between columns; The lines position interaction was not significant. That is, regardless of the row or column used was coincidence in the lines performance;
28 Results and Discussion FC Virginia Burley Figure 1. Curvature of the coefficient of variation (CV%) depending on the size of the plot (a) and point of maximum curvature (b). For cured leaves weight (g/plant), sugar content (%) and alkaloid content (%), Burley variety groups (top) and Virginia (below). Rio Negro, Parana, 2009/2010.
29 Conclusions The use of border is not justified because the interactions between lines position in the plots was not significant, showing that the behavior of the lines was similar in the different positions; The performance of the plants varied with the position column in the plot; The experimental precision evaluated by CV% increased with increasing the plot size, however, the method of maximum curvature of the coefficient of variation showed no significant increases in accuracy if the number of plants is greater than seven; The ideal number of plants for a tobacco plot would be larger than seven. The ideal plots would be formed by two rows of plants.
30 Siyabonga Muito Obrigado Many Thanks Alah hamdulila alah akbar Grazie mille Arigatougozaimasu बह त बह त धन यव Asante sana 非常感謝 Tack så mycket Terima kasih banyak Xie Xie ne Cảm ơn bạn rất nhiều Je vous remercie beaucoup Vielen Dank Dankie Çok teşekkür ederim
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