# Effect of Planting Material Type on Experimental Trial Quality and Performance Ranking of Sugarcane Genotypes

Authors: Michel Choairy de Moraes; Ana Carolina Ribeiro Guimarães; Dilermando Perecin; Manuel Benito Sainz
Journal: International Journal of Agronomy
 (2018)

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## Abstract

In recent years, the use of presprouted setts (MPB, which stands for “mudas pre-brotadas” in Portuguese) to establish commercial sugarcane nurseries has grown in Brazil. MPB and single-bud setts (SBS) have the advantage of requiring less planting material and enabling a higher multiplication rate of the source material as compared with the conventional multibud sett (MBS) planting system. Sugarcane breeding programs could also potentially benefit from the precise spacing afforded by MPB or SBS planting materials, by reducing trial variability. However, the effect of planting material type on the performance ranking and consequent selection of sugarcane clones in a breeding program has not been previously investigated. We present results on possible interactions between genotype and the type of planting material (MPB, MBS, or SBS) on key performance parameters, like sugar content, cane yield, and sugar yield, in the context of the intermediate phase of a sugarcane breeding program. Our results indicate that trial quality does not necessarily improve with the use of MPB or SBS planting materials and that type of planting material has a significant effect on the ranking of sugarcane genotypes, and this needs to be taken into consideration when considering the use of new planting technologies in breeding trials of vegetatively propagated crops such as sugarcane.


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## Body

## 1. Introduction

Sugarcane is a vegetatively propagated crop. Manual planting of multibud setts (MBS) is the traditional planting material used in the planting of commercial nurseries and production fields. To minimize the risk of gaps in the resultant stand, manual planting rates are high (15–21 buds/meter), corresponding to 11–14 tonnes (T) of planting material/hectare (ha). With mechanized planting, the amount of planting material used is even larger, reaching levels greater than 20 T/ha. Sugarcane production costs have increased due to increased labor and agricultural input costs, with the cost of planting material accounting for almost 25% of operational production costs [1]. The large quantity of planting material required in traditional planting systems also leads to problems with logistics, storage, and loss of bud viability.New planting systems have been developed to overcome some of the disadvantages of traditional methods. The presprouted seedling (MPB) planting system allows for a reduction in the quantity of planting material and better control of seedling vigor [2–6]. Another planting system (Plene™) developed by Syngenta uses 5 cm, single-bud setts (SBS) treated with a pesticidal slurry [7]. Bud chips are also a promising alternative for reduction of sugarcane production costs, although improvement of survival rates and plant vigor under field conditions is needed [8].Genetic improvement of sugarcane is based on the selection and cloning of superior genotypes of segregating populations obtained through sexual crosses between different individuals [9]. Different methodologies have been used in the selection of individuals in the early stages of sugarcane breeding: mass selection [9], Australian sequential selection [10], modified sequential selection [11], and individual simulated best linear unbiased prediction (BLUP) [12, 13], among them. After the initial seedling phase, selection is based on planting clonal setts, as MBS, in plots that are assessed for key agronomic attributes. As selection progresses, the size of the plot and the number of replicates per clone increase, which allows better assessment of the materials under selection.Sugarcane breeding trials could benefit from using planting systems that increase efficiency and data quality. Higher trial data quality could result from more precise spacing in trial plots. Planting systems with the potential to do more replications in early breeding stages due to more efficient utilization of scarce planting material would provide an additional benefit. However, planting system modifications have the potential to affect the ranking and consequent selection of genotypes, since experimental results of important performance attributes such as yield and sugar content could differ depending on the planting system used. Studies on intrarow spacing and number of buds per sett in commercial varieties [14] and optimal planting rates using whole stalks for different varieties have been reported [15]. However, to date and to our knowledge, there have been no reports in the literature about the effect of presprouted seedling (MPB) or single-bud setts planting materials on clonal selection in sugarcane breeding programs.In the present study, we evaluate potential interactions between genotypes and the type of planting system and whether the type of planting material has an effect on trial data quality.


## 2. Materials and Methods

### 2.1. Trial Design

Three types of planting material were tested: 3-4 bud setts (MBS—conventional method), presprouted seedlings (MPB), and 5 cm, single-bud setts (SBS). All planting materials were generated from selected healthy stalks harvested approximately 9 months after planting.All three types of planting material were planted at a single location in three different adjacent trials. All trials were in a randomized complete block design with 3 replicates. Plots consisted of two 10 m rows spaced 1.5 m apart. Adjacent plots were spaced 3 m apart along their length.The same genotypes (cultivars and clones; see Table1) were planted in each of the 3 different trials, adjacent to each other. Clones were part of the 3rd stage of the selection process of the Syngenta sugarcane breeding program. This stage is the first one in which replicates are used and plot weight is directly measured following mechanical harvesting.Table 1
Varieties and clones used in the study.
Varieties
Clones
RB86-7515
S09-0001
S09-0040
S09-0114
RB96-6928
S09-0007
S09-0046
S09-0122
SP81-3250
S09-0011
S09-0048
S09-0140
S09-0022
S09-0052
S09-0144
S09-0023
S09-0055
S09-0146
S09-0031
S09-0069
S09-0148
S09-0036
S09-0080
S09-0153
S09-0037
S09-0081
S09-0154
S09-0038
S09-0098The process for making and planting the different types of planting materials is described below and in the accompanying figures (Figures1–3). In all cases, supplemental irrigation was applied until the crop stand was well established.Figure 1
Process for SBS (single-bud sett) production and planting.Figure 2
Process for MPB (presprouted seedling) production and planting.Figure 3
Process for MBS (multibud sett) production and planting.


#### 2.1.1. Presprouted Setts (MPB)

Eight-month-old sugarcane stalks were harvested and 5 cm, single-bud setts were cut and planted in soil mix. The resultant sprouted seedlings were manually planted in the field trial after 50 days at an intrarow spacing of 0.5 m.


#### 2.1.2. Single-Bud Setts (SBS)

Ten-month-old sugarcane stalks were harvested, and 5-cm, single-bud setts were cut and treated with a slurry consisting of industrial proprietary treatment. Subsequently, these were planted manually in the field at a rate of 8 single-bud setts per meter.


#### 2.1.3. Multibud Setts (MBS): Conventional Method

Ten-month-old sugarcane stalks were harvested manually and placed in row furrows. These were cut with a machete in the furrow into 30–40 cm pieces, as per conventional manual cane planting practice.The 3 field trials were planted in a single week in April 2014. Fertilization and cultural practices followed conventional commercial practice and were the same for all trials. Evaluations of sugarcane agronomic parameters were made over two harvest cycles (plant cane and 1st ratoon). In August 2015, a sample of 10 stalks per plot was subjected to laboratory POL analysis (a measure of sugar content). Subsequently, in the same month, the trial was mechanically harvested and whole, individual plots were weighed to estimate TCH (tonnes cane per hectare). From the POL and TCH parameters, the TPH (tonnes POL per hectare) was calculated for the plant cane harvest. It was not possible to measure POL in the 1st ratoon harvest, but in May 2016, the Brix of 5 stalks per plot was taken and averaged. Mechanized harvesting and weighing of trial plots were conducted in June 2016. Thus, Brix, TCH, and TBH parameters (tonnes Brix per hectare) were estimated for the 1st ratoon harvest.


### 2.2. Statistical Analysis

Analysis of variance was done by planting material (MPB, SBS, and MBS) and harvest cycle (plant cane, 1st ratoon), considering the effects of genotype (26 clones and three varieties) and blocks (3 per planting material). For each of the three traits (POL or Brix; TCH; and TPH or TBH), the ratio between the largest and smallest mean residual squares was less than three. As a result, we performed joint analyses by traditional ANOVA and mixed model restricted maximum likelihood (REML)/best linear unbiased prediction (BLUP), in which planting material, harvest cycle, and blocks nested within planting material were fixed effects, and genotype, as well as genotypic interactions with planting material and harvest cycle, were random effects. A split-plot design was not used due to possible plot border effects resulting from different growth rates of different adjacent plant material types. As stated above, mean residual squares within each trial were similar; consequently, we performed a joint analysis by ANOVA (despite the absence of randomization), much as experiments across locations are analyzed.Within each cycle (plant and ratoon cane), statistical modeling was done as for a split-plot design in time.


## 2.1. Trial Design

Three types of planting material were tested: 3-4 bud setts (MBS—conventional method), presprouted seedlings (MPB), and 5 cm, single-bud setts (SBS). All planting materials were generated from selected healthy stalks harvested approximately 9 months after planting.All three types of planting material were planted at a single location in three different adjacent trials. All trials were in a randomized complete block design with 3 replicates. Plots consisted of two 10 m rows spaced 1.5 m apart. Adjacent plots were spaced 3 m apart along their length.The same genotypes (cultivars and clones; see Table1) were planted in each of the 3 different trials, adjacent to each other. Clones were part of the 3rd stage of the selection process of the Syngenta sugarcane breeding program. This stage is the first one in which replicates are used and plot weight is directly measured following mechanical harvesting.Table 1
Varieties and clones used in the study.
Varieties
Clones
RB86-7515
S09-0001
S09-0040
S09-0114
RB96-6928
S09-0007
S09-0046
S09-0122
SP81-3250
S09-0011
S09-0048
S09-0140
S09-0022
S09-0052
S09-0144
S09-0023
S09-0055
S09-0146
S09-0031
S09-0069
S09-0148
S09-0036
S09-0080
S09-0153
S09-0037
S09-0081
S09-0154
S09-0038
S09-0098The process for making and planting the different types of planting materials is described below and in the accompanying figures (Figures1–3). In all cases, supplemental irrigation was applied until the crop stand was well established.Figure 1
Process for SBS (single-bud sett) production and planting.Figure 2
Process for MPB (presprouted seedling) production and planting.Figure 3
Process for MBS (multibud sett) production and planting.


### 2.1.1. Presprouted Setts (MPB)

Eight-month-old sugarcane stalks were harvested and 5 cm, single-bud setts were cut and planted in soil mix. The resultant sprouted seedlings were manually planted in the field trial after 50 days at an intrarow spacing of 0.5 m.


### 2.1.2. Single-Bud Setts (SBS)

Ten-month-old sugarcane stalks were harvested, and 5-cm, single-bud setts were cut and treated with a slurry consisting of industrial proprietary treatment. Subsequently, these were planted manually in the field at a rate of 8 single-bud setts per meter.


### 2.1.3. Multibud Setts (MBS): Conventional Method

Ten-month-old sugarcane stalks were harvested manually and placed in row furrows. These were cut with a machete in the furrow into 30–40 cm pieces, as per conventional manual cane planting practice.The 3 field trials were planted in a single week in April 2014. Fertilization and cultural practices followed conventional commercial practice and were the same for all trials. Evaluations of sugarcane agronomic parameters were made over two harvest cycles (plant cane and 1st ratoon). In August 2015, a sample of 10 stalks per plot was subjected to laboratory POL analysis (a measure of sugar content). Subsequently, in the same month, the trial was mechanically harvested and whole, individual plots were weighed to estimate TCH (tonnes cane per hectare). From the POL and TCH parameters, the TPH (tonnes POL per hectare) was calculated for the plant cane harvest. It was not possible to measure POL in the 1st ratoon harvest, but in May 2016, the Brix of 5 stalks per plot was taken and averaged. Mechanized harvesting and weighing of trial plots were conducted in June 2016. Thus, Brix, TCH, and TBH parameters (tonnes Brix per hectare) were estimated for the 1st ratoon harvest.


## 2.1.1. Presprouted Setts (MPB)

Eight-month-old sugarcane stalks were harvested and 5 cm, single-bud setts were cut and planted in soil mix. The resultant sprouted seedlings were manually planted in the field trial after 50 days at an intrarow spacing of 0.5 m.


## 2.1.2. Single-Bud Setts (SBS)

Ten-month-old sugarcane stalks were harvested, and 5-cm, single-bud setts were cut and treated with a slurry consisting of industrial proprietary treatment. Subsequently, these were planted manually in the field at a rate of 8 single-bud setts per meter.


## 2.1.3. Multibud Setts (MBS): Conventional Method

Ten-month-old sugarcane stalks were harvested manually and placed in row furrows. These were cut with a machete in the furrow into 30–40 cm pieces, as per conventional manual cane planting practice.The 3 field trials were planted in a single week in April 2014. Fertilization and cultural practices followed conventional commercial practice and were the same for all trials. Evaluations of sugarcane agronomic parameters were made over two harvest cycles (plant cane and 1st ratoon). In August 2015, a sample of 10 stalks per plot was subjected to laboratory POL analysis (a measure of sugar content). Subsequently, in the same month, the trial was mechanically harvested and whole, individual plots were weighed to estimate TCH (tonnes cane per hectare). From the POL and TCH parameters, the TPH (tonnes POL per hectare) was calculated for the plant cane harvest. It was not possible to measure POL in the 1st ratoon harvest, but in May 2016, the Brix of 5 stalks per plot was taken and averaged. Mechanized harvesting and weighing of trial plots were conducted in June 2016. Thus, Brix, TCH, and TBH parameters (tonnes Brix per hectare) were estimated for the 1st ratoon harvest.


## 2.2. Statistical Analysis

Analysis of variance was done by planting material (MPB, SBS, and MBS) and harvest cycle (plant cane, 1st ratoon), considering the effects of genotype (26 clones and three varieties) and blocks (3 per planting material). For each of the three traits (POL or Brix; TCH; and TPH or TBH), the ratio between the largest and smallest mean residual squares was less than three. As a result, we performed joint analyses by traditional ANOVA and mixed model restricted maximum likelihood (REML)/best linear unbiased prediction (BLUP), in which planting material, harvest cycle, and blocks nested within planting material were fixed effects, and genotype, as well as genotypic interactions with planting material and harvest cycle, were random effects. A split-plot design was not used due to possible plot border effects resulting from different growth rates of different adjacent plant material types. As stated above, mean residual squares within each trial were similar; consequently, we performed a joint analysis by ANOVA (despite the absence of randomization), much as experiments across locations are analyzed.Within each cycle (plant and ratoon cane), statistical modeling was done as for a split-plot design in time.


## 3. Results

Mean values of TPH_TBH were similar for the three planting material types tested (Table2). Average POL and Brix were higher for SBS and MPB than for conventional MBS planting material, whereas average TCH for SBS was lower than that of the other two types of planting material. Average TCH for MPB and the conventional MBS planting material was quite similar, as previously observed by [16], who noted that MPB required less planting material. Regarding trial coefficients of variation (CVs), these were similar across trials, planting material type, and harvest cycles, being in general lower for POL_Brix. TCH CVs for MPB and SBS were slightly higher than for the conventional MBS planting type. For TPH_TBH, the plant cane average was higher for MPB, but no differences between the 3 planting material types were observed in the 1st ratoon. The CV for TBH was slightly lower in the conventional MBS planting type than for the other two.Table 2
Average performance parameters and coefficients of variation (CV) in trials with 3 types of planting materials, 29 genotypes, and 3 blocks (replicates) per planting material.
Performance parameter
MBS average
SBS average
MPB average
POL-Brix
Plant cane POL
12.42
13.55
13.05
1st ratoon Brix
13.59
14.71
14.71
%CV, plant cane POL
7.47
5.89
7.34
%CV, 1st ratoon Brix
8.25
7.23
7.46
TCH
Plant cane TCH
136.11
134.78
139.10
1st ratoon TCH
104.79
97.35
106.95
%CV, plant cane TCH
12.54
15.15
13.95
%CV, 1st ratoon TCH
12.19
16.69
14.29
TPH_TBH
Plant cane TPH
16.85
18.10
17.94
1st ratoon TBH
14.29
14.21
14.84
%CV, plant cane TPH
14.44
17.01
16.85
%CV, 1st ratoon TBH
15.81
15.53
16.81Conventional analysis of variance (Table3) points to highly significant effects of genotype (p<0.0001) and of the interaction of genotype with planting material type (p≤0.1) across the different parameters evaluated, indicating that rank ordering of genotypes could be affected depending on the planting material type used in the trial. However, harvest cycle and the interaction of harvest cycle with genotype also had a highly significant effect (p<0.0001) on the parameters tested.Table 3
Mean squares (MS) andp values pooled joint analysis of variance. Conventional fixed model, with planting material type as test and harvest cycle (plant cane, 1st ratoon) as split-plot, was used.
Source of variation
POL_BRIX
TCH
TPH_TBH
Planting material (PM)
59.60 (p<0.0001)
1484.92 (p=0.02)
24.17(p=0.06)
Genotype
19.83 (p<0.0001)
4855.15 (p<0.0001)
104.95 (p<0.0001)
Genotype × PM
1.48 (p=0.01)
494.80 (p=0.07)
12.48 (p=0.03)
Harvest cycle
141.37 (p<0.0001)
12,2717.39 (p<0.0001)
1085.90 (p<0.0001)
Harvest cycle × PM
1.88 (p=0.22)
364.55 (p=0.17)
13.61 (p=0.08)
Harvest cycle × genotype
3.87 (p<0.0001)
1002.86 (p<0.0001)
16.51 (p<0.0001)Random effect variances obtained using a mixed model (Table4) show that genotype accounts for approximately 36–37% of the variability, which allows for selection of the best clones in a breeding program. Interactions of genotype with planting material type ranged from 2.7% to 7.1%: lower but significant (p<0.05) for POL_Brix and higher for TCH and TPH_TBH, indicating that planting material type has the potential to affect the rank order of clones in the selection process. Interaction of genotype with harvest cycle was significant (p<0.05), as expected, since varieties in commercial sugarcane fields will also differ in their performance depending on harvest cycle that includes the environmental effects of growing season. There is also a weak interaction of harvest cycle with planting material type (p<0.25; data not shown).Table 4
Components and percentage of variance as obtained in a pooled joint analysis with a mixed REML/BLUP model, considering genotype, genotype × planting material, genotype × harvest cycle, and residual as random effects.
Variable
POL_BRIX
TCH
TPH_TBH
Genotypes
0.8658 (37.08%)
232.02 (36.41%)
5.1067 (36.16%)
Genotypes × PM
0.0632 (2.71%)
37.75 (5.92%)
1.0028 (7.10%)
Genotypes × harvest cycle
0.3081 (13.20%)
93.47 (14.67%)
1.3193 (9.34%)
Residual
1.0978 (47.02%)
274.07 (43.00%)
6.6918 (47.39%)Predicted genotypic differences (PGDs) obtained in the mixed model (Tables5 to 7) are presented to show differences in genotype ranking for each performance parameter, by planting material type. For each genotype, the expected genotypic value is the mean + PGD.Table 5
Predicted genotypic differences (PGD) for TPH_TBH across 29 genotypes (mean plant cane TPH and 1st ratoon TBH) and three planting material types (MBS, SBS, or MPB).
Rank
Genotype
MBS
p value
Genotype
SBS
p value
Genotype
MPB
p value
1
S09-0146
4.7444
<0.001
RB966928
4.4965
<0.001
RB966928
4.592
0.0002
2
RB867515
4.4646
<0.001
S09-0144
4.1737
0.0002
RB867515
4.5099
0.0003
3
RB966928
3.702
0.001
S09-0146
4.1348
0.0003
S09-0122
3.0073
0.0095
4
S09-0031
3.0899
0.006
S09-0114
3.4487
0.0021
S09-0114
2.614
0.033
5
S09-0114
2.4153
0.031
S09-0038
3.2957
0.0384
S09-0140
2.5809
0.0254
6
S09-0038
2.0233
0.07
S09-0052
2.7208
0.0325
S09-0144
2.304
0.0597
7
S09-0052
1.887
0.09
S09-0153
2.1413
0.0919
S09-0031
1.773
0.1228
8
S09-0140
1.6314
0.143
S09-0140
2.0738
0.0607
S09-0069
1.5078
0.1889
9
S09-0144
1.2664
0.254
S09-0148
1.6054
0.1454
S09-0023
1.3682
0.2614
10
S09-0148
1.2593
0.257
RB867515
0.8213
0.4548
S09-0052
1.0307
0.3682
11
S09-0037
0.7224
0.515
S09-0037
0.7444
0.555
S09-0146
0.9092
0.4548
12
S09-0122
0.3986
0.719
S09-0031
0.3428
0.7548
S09-0007
0.5837
0.6099
13
SP813250
0.3901
0.725
S09-0069
0.1064
0.9227
S09-0153
0.0999
0.9345
14
S09-0046
0.2438
0.826
S09-0055
−0.045
0.9713
S09-0046
−0.209
0.8549
15
S09-0023
0.1558
0.888
S09-0122
−0.09
0.9431
S09-0148
−0.643
0.6231
16
S09-0055
−0.394
0.722
SP813250
−0.183
0.8674
S09-0055
−0.699
0.5416
17
S09-0069
−0.435
0.695
S09-0154
−1.439
0.1916
S09-0038
−0.912
0.4259
18
S09-0153
−0.659
0.552
S09-0080
−1.678
0.1282
S09-0040
−0.943
0.4101
19
S09-0007
−0.694
0.532
S09-0022
−1.793
0.1565
S09-0001
−1.12
0.3576
20
S09-0036
−0.708
0.524
S09-0081
−1.854
0.0932
S09-0048
−1.192
0.2983
21
S09-0022
−1.703
0.126
S09-0040
−2.074
0.0607
S09-0037
−1.501
0.218
22
S09-0001
−2.118
0.058
S09-0023
−2.299
0.0379
S09-0081
−1.512
0.2148
23
S09-0154
−2.368
0.034
S09-0046
−2.327
0.1421
S09-0036
−1.605
0.1621
24
S09-0048
−2.389
0.033
S09-0007
−2.401
0.0303
S09-0022
−1.733
0.1313
25
S09-0081
−2.808
0.012
S09-0036
−2.527
0.0228
SP813250
−2.164
0.0767
26
S09-0098
−2.91
0.01
S09-0011
−2.685
0.0157
S09-0154
−2.295
0.0464
27
S09-0011
−3.422
0.002
S09-0048
−2.789
0.0285
S09-0098
−2.682
0.0203
28
S09-0080
−3.856
0.0007
S09-0001
−2.907
0.0226
S09-0080
−3.669
0.0017
29
S09-0040
−3.93
0.0005
S09−0098
−3.015
0.0068
S09-0011
−4.001
0.0013
TM1
—
15.57B
—
—
16.16A
—
—
16.26A
—
1TM with different letters indicate significant (p value < 0.05) by T-test (LSD). TM = trial mean; p value testing PGD as null.Table 6
Predicted genotypic differences (PGD) for TCH across 29 genotypes (mean plant cane and 1st ratoon TCH) and three planting material types (MBS, SBS, or MPB).
Rank
Genotype
MBS
p value
Genotype
SBS
p value
Genotype
MPB
p value
1
S09-0146
39.089
<0.001
S09-0146
26.489
0.0008
RB867515
28.893
0.0003
2
RB867515
29.224
<0.001
RB966928
24.382
0.0019
RB966928
22.466
0.0046
3
S09-0114
20.817
0.006
S09-0114
23.463
0.0028
S09-0114
16.734
0.0337
4
S09-0038
20.557
0.006
S09-0153
22.079
0.0138
S09-0146
15.104
0.0548
5
RB966928
19.358
0.01
S09-0038
21.821
0.049
S09-0007
14.768
0.0461
6
S09-0031
14.071
0.06
S09-0144
21.342
0.0063
S09-0069
13.874
0.0608
7
S09-0144
12.829
0.086
S09-0052
16.24
0.0677
S09-0154
10.657
0.1486
8
S09-0052
8.2784
0.266
S09-0154
12.845
0.097
S09-0140
9.5021
0.1974
9
S09-0154
5.8661
0.43
RB867515
8.122
0.2923
S09-0038
9.2683
0.2086
10
S09-0153
5.3894
0.469
S09-0140
6.2557
0.4168
S09-0023
8.135
0.2985
11
S09-0007
4.4072
0.553
S09-0148
5.5204
0.4735
S09-0040
7.6597
0.2982
12
S09-0140
3.4683
0.641
S09-0069
3.8803
0.6142
S09-0153
6.2699
0.4226
13
S09-0148
3.4538
0.642
S09-0037
1.7567
0.8423
S09-0144
6.254
0.4238
14
SP813250
2.9916
0.687
S09-0040
0.8263
0.9145
S09-0122
4.4561
0.5445
15
S09-0069
0.5793
0.938
S09-0055
−0.536
0.9516
S09-0031
4.0024
0.5862
16
S09-0037
0.0882
0.991
SP813250
−2.002
0.7948
S09-0052
−0.356
0.9613
17
S09-0023
−2.079
0.7800
S09-0031
−2.242
0.7708
S09-0148
−1.780
0.8324
18
S09-0040
−2.310
0.756
S09-0122
−4.794
0.5873
S09-0001
−5.042
0.5189
19
S09-0036
−4.000
0.591
S09-0022
−6.746
0.4453
S09-0036
−7.231
0.326
20
S09-0122
−7.842
0.292
S09-0080
−8.124
0.2922
SP813250
−8.281
0.29
21
S09-0055
−9.301
0.212
S09-0001
−11.39
0.1986
S09-0055
−10.43
0.1572
22
S09-0022
−11.24
0.132
S09-0007
−12.61
0.1033
S09-0046
−11.11
0.1323
23
S09-0046
−11.71
0.117
S09-0081
−14.50
0.0614
S09-0037
−11.96
0.1274
24
S09-0001
−12.44
0.096
S09-0023
−20.16
0.0098
S09-0048
−14.78
0.046
25
S09-0098
−20.84
0.006
S09-0011
−21.57
0.0058
S09-0022
−15.45
0.0371
26
S09-0081
−23.99
0.002
S09-0036
−21.81
0.0053
S09-0098
−17.09
0.0214
27
S09-0048
−24.47
0.001
S09-0098
−22.26
0.0045
S09-0081
−18.45
0.0194
28
S09-0011
−28.56
2E−04
S09-0048
−22.57
0.0117
S09−0080
−25.76
0.0006
29
S09-0080
−31.69
<0.001
S09-0046
−23.71
0.0327
S09-0011
−30.31
0.0002
TM1
—
120.45A
—
—
116.07B
—
—
121.62A
—
1TM with different letters indicate significant (p value<0.05) by T-test (LSD). TM = trial mean; p value testing PGD as null.Table 7
Predicted genotypic differences (PGD) for POL_Brix across 29 genotypes (mean plant cane POL and 1st ratoon Brix) and three planting material types (MBS, SBS, or MPB). TM = trial mean;p value testing PGD as null.
Rank
Genotype
MBS
p value
Genotype
SBS
p value
Genotype
MPB
p value
1
S09-0046
1.4759
0.0021
S09-0046
1.7214
<0.001
S09-0122
1.8558
0.0002
2
S09-0122
1.2755
0.0076
S09-0140
0.9188
0.0279
S09-0144
1.1144
0.0211
3
S09-0140
0.8705
0.0667
S09-0144
0.7226
0.0828
S09-0046
1.1046
0.0222
4
S09-0031
0.8431
0.0757
S09-0052
0.7143
0.0864
S09-0140
0.9496
0.0488
5
RB966928
0.8321
0.0795
S09-0122
0.6489
0.119
S09-0081
0.9468
0.0495
6
S09-0055
0.7827
0.0989
S09-0036
0.6434
0.1222
RB966928
0.9301
0.0536
7
S09-0148
0.7676
0.1055
S09-0023
0.6406
0.1238
S09-0031
0.9022
0.0611
8
S09-0048
0.7429
0.1171
S09-0148
0.5182
0.2125
S09-0052
0.8589
0.0744
9
S09-0052
0.7291
0.124
RB966928
0.489
0.2392
S09-0048
0.6453
0.179
10
S09-0037
0.6221
0.1889
S09-0031
0.4834
0.2446
S09-0055
0.6145
0.2005
11
RB867515
0.3585
0.448
S09-0011
0.3832
0.3558
RB867515
0.44
0.3587
12
S09-0023
0.3516
0.4567
S09-0048
0.3053
0.4617
S09-0011
0.3297
0.4913
13
S09-0081
0.202
0.6687
S09-0037
0.2511
0.5449
S09-0037
0.2697
0.5734
14
S09-0080
0.154
0.7443
S09-0055
0.2191
0.5972
S09-0148
0.1943
0.6849
15
S09-0011
0.1306
0.782
S09-0114
0.2038
0.623
S09-0022
0.1859
0.6978
16
S09-0114
-0.1494
0.7517
SP813250
0.1482
0.7208
S09-0023
0.1105
0.8175
17
S09-0146
−0.1549
0.7429
S09-0081
0.1384
0.7384
S09-0114
0.01136
0.9811
18
SP813250
−0.1549
0.7429
S09-0146
0.137
0.741
S09-0069
−0.2428
0.6122
19
S09-0036
−0.1919
0.6843
S09-0098
−0.1356
0.7436
S09-0001
−0.3237
0.4992
20
S09-0022
−0.2455
0.6032
S09-0038
−0.1982
0.6326
S09-0080
−0.3433
0.4737
21
S09-0098
−0.2798
0.5536
RB867515
−0.2858
0.4908
S09-0098
−0.5695
0.2353
22
S09-0144
−0.3567
0.4503
S09-0069
−0.4527
0.2757
S09-0153
−0.6351
0.1859
23
S09-0069
−0.439
0.353
S09-0022
−0.5598
0.1781
S09-0146
−0.691
0.1504
24
S09-0038
−0.4733
0.3168
S09-0080
−0.5765
0.1656
S09-0036
−0.751
0.1183
25
S09-0001
−0.542
0.2519
S09-0153
−0.7323
0.0789
SP813250
−0.8515
0.0769
26
S09-0153
−0.9648
0.0424
S09-0007
−0.7852
0.0597
S09-0007
−1.1126
0.0213
27
S09-0007
−1.2119
0.0111
S09-0001
−1.272
0.0025
S09-0040
−1.4477
0.0029
28
S09-0154
−2.1933
<0.001
S09-0040
−1.7185
<0.001
S09-0038
−1.7828
0.0003
29
S09-0040
−2.7808
<0.001
S09-0154
−2.5698
<0.001
S09-0154
−2.7127
—
TM1
—
13.00C
—
—
14.13A
—
—
13.45B
—
1TM with different letters indicate significant (p value<0.05) by T-test (LSD).For TPH_TBH (Table5), the MPB and SBS planting materials produced similar means (p<0.05) that were higher than those obtained with the MBS conventional planting material. Results with the 10 best genotypes show differences in rank ordering. For example, using conventional MBS planting material, the best clone was S09-0146, yielding 4.74 tonnes above the 15.57 TPH_TBH general average, and slightly surpassing the two RB variety checks. S09-0146 is ranked 3rd for TPH_TBH when planted as SBS, but is not among the top 10 genotypes when MPB is used as the planting material. Clone S09-0031 is ranked 4th just below the two RB checks when planted as MBS and is in the top 10 when MPB planting material is used, but not when planted as SBS. Similarly, S09-0122 is the top ranked clone when MPB planting material is used, suggesting good adaptation to this planting methodology, but is not in the top 10 when using the other planting methodologies. Among the checks, RB966928 was ranked 3rd when conventional MBS planting material is used, but 1st when MPB or SBS planting materials are used, whereas RB867515 did not perform well when SBS planting material was used.For TCH (Table6), the highest average yields were obtained from conventional MBS and MPB planting materials, with SBS planting material having a significantly lower (p<0.05) mean. In terms of rank, MPB and SBS methods improved the performance of check variety RB966928 while SBS leads to deterioration in performance of the RB867515 check. Among the clones, S09-0146 is again at the 1st place when conventional MBS and SBS planting materials are used, but at the 4th place using MPB, whereas clone S09-114 ranks 3rd with all 3 planting material types. The clone S09-0038, at the 4th and 5th place when planted as MBS or SBS, respectively, drops to the 9th place when MPB is used, whereas the clone S0-007, at the 4th place when the MPB planting material is used, drops to 11th with MBS and 22nd with SBS.For POL and Brix (Table7), means were significantly different (p<0.05) depending on the type of planting material, being higher (14.13) when SBS planting material was used; intermediate for MPB (13.45); and lowest for conventional MBS (13.00). Four clones showed higher POL_Brix than the RB966928 check: clones S09-0046 and S9-0122 ranked highest in POL_Brix but were not among the top 10 for TCH. On the other hand, clone S09-0146 which ranked high for TCH was not among the best clones for POL_Brix.Results of the correlations between predicted genotypic differences (PDG; Tables5 to 7) were used to evaluate correlations between the 3 different planting material types for the three performance parameters (Table 8). Correlations of 0.82–0.85 were observed when comparing conventional MBS and the MPB and SBS planting materials for POL_Brix, and when comparing conventional MBS with MPB for TCH. Slightly lower correlations (0.75–0.79) were observed when comparing conventional MBS with MPB and SBS for TPH_TBH, and for conventional MBS and SBS for TCH. Hence, there are relatively good but not perfect correlations between the conventional MBS and the SBS and MPB planting materials, possibly due to interactions between some genotypes and planting material type. Correlations between MPB and SBS were generally lower, ranging from 0.61 for TPH_TBH, 0.65 for TCH, and 0.76 for POL_Brix, possibly due to higher risks to stand establishment when using SBS planting materials.Table 8
Pearson’s correlation coefficients between average performance parameters for 29 genotypes across three different planting material types, using predicted genotypic differences (PDG) from Tables5 to 7, for each performance parameter.
Performance parameter
MBS
MPB
SBS
POL_Brix
MBS
—
0.82
0.85
MPB
—
—
0.75
TCH
MBS
—
0.85
0.79
MPB
—
—
0.65
TPH_TBH
MBS
—
0.77
0.78
MPB
—
—
0.61
N = 29 for all correlations (Prob > (r) < 0.05 under rho = 0.


## 4. Discussion

The observed CVs for the parameters analyzed (Table2) were in line with those previously observed in the scientific literature. Couto et al. [17] evaluated the range of CVs observed in sugarcane experiments and concluded that the TCH and TPH parameters presented the highest CV ranges, whereas % sucrose presented the lowest. These authors also indicated that upper CV limits for good to high precision as being 10%, 15%, and 19% for % sucrose, TCH, and TPH, respectively. Using these numbers as a guide, in the present study, only the TCH in the SBS trial is higher than these limits, which indicates that trial quality was high enough to draw conclusions from this study. Although we observe lower POL and Brix CVs with the SBS and MPB planting material (Table 2), in general our data do not provide evidence that MPB or SBS planting materials decrease variability, and thus improve trial quality, in sugarcane breeding selection trials.Conventional analysis of variance (Table3) points to highly significant effects of genotype and harvest cycle, and also of the interaction of harvest cycle with planting material. Milligan et al. [18] also found that genotype by harvest cycle interaction was important for sugar yield and its component traits. In this study, the pooled variation due to genotype (Table 4) was found to be sufficient to select the top genotypes to advance to the next stage. We found a weaker but still significant interaction (p≤0.1) of genotype by planting material, suggesting that the rank ordering of genotypes could be affected depending on the planting material type used in the trial. In contrast, planting material alone had a highly significant effect only on variation of POL_Brix. These results were verified by the differences in the rank ordering of clones we observed the TPH_TBH, TCH, and POL_Brix performance parameters (Tables 5–7).Changes in rank order of genotypes can impact the effectiveness of clonal selection in sugarcane breeding. For example, the TPH_TBH parameter can be considered the most important one for selection in the intermediate phases of a program. Selection indices in different breeding stages vary by breeding program, as shown by [19]. In our breeding program, a 10 to 20% selection index was used in the third stage. A selection index of 20% from the trials described in this study, and considering data from 2 harvest cycles, would advance 6 clones to the next phase. If we consider conventional MBS planting material as the most relevant to the current commercial sugarcane planting practice, clones S09-0146, S09-0031, S09-0114, S09-0038, S09-0052, and S09-0140 would then be selected. However, of these 6 clones, only 4 (S09-0146, S09-0114, S09-0038, and S09-0052) would be advanced if using the SBS planting material, and only 3 (S09-0114, S09-0140, and S09-0031) would be advanced by using MPB planting material.Orgeron et al. [15], studying whole stalks planting rating effect in 8 different genotypes, found no planting rate by genotype interaction for cane and sugar yield. Similarly, Netsanet and Tegene [14], comparing three different commercial varieties and their behavior in terms of intrarow spacing and buds per setts, found no significant interaction effect or spacing used on sugar and cane yield. On the contrary, what we found in the present study was that planting material type by genotype interaction is significant. The different conclusions between the studies possibly are due to the higher number of genotypes, and the genotypes per se, used in the present study and due to the different planting material used (MPB and SBS), compared with the other cited studies.On an average, good correlations of performance parameter were observed between the different types of planting material. These correlations mask the significant effect on the ranking of individual clones in these trials. MPB and SBS planting methodologies have generated enormous interest in the Brazilian sugarcane industry and have undeniable advantages in terms of reduction of planting costs and material handling logistics. However, the use of these new planting methodologies in a sugarcane breeding program will result in likely selection of genotypes well adapted to the particular type of planting material used, but may not have the best agronomic performance if used in commercial plantings using other planting systems, such as the current conventional MBS planting system.Our study indicates that trial quality does not necessarily improve with the use of MPB or SBS planting materials compared with the conventional MBS. Additionally, the type of planting material has a significant effect on the ranking of sugarcane genotypes. Because of that, when considering the use of new planting technologies in breeding trials of sugarcane, this needs to be taken into consideration for the selection of genotypes for cane yield and sugar parameters.



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*Source: 3723471-2018-09-02.xml*