# Biometrics Assessment of Cluster- and Berry-Related Traits of Muscadine Grape Population

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## Abstract

**:**

## 1. Introduction

## 2. Results

#### 2.1. Determination of Flower Structure (FLS) in Muscadine Population

#### 2.2. Cluster-Related Traits

#### 2.2.1. Cluster Length

#### 2.2.2. Cluster Width

#### 2.2.3. Cluster Weight

#### 2.2.4. Number of Berries/Cluster

#### 2.2.5. Cluster Compactness

#### 2.2.6. Frequency Distribution of Cluster-Related Traits

#### 2.3. Berry-Related Traits

#### 2.3.1. Berry Length

#### 2.3.2. Berry Width

#### 2.3.3. Berry Weight

#### 2.3.4. Number of Seeds/Berry (N.S/B)

#### 2.3.5. Weight of Seeds/Berry (W.S/B)

#### 2.3.6. Berry Firmness

#### 2.3.7. Scar Pattern (SP)

#### 2.4. Frequency Distribution of Berry-Related Traits

#### 2.5. Classification of Muscadine Genotypes Based on the Evaluated Traits

#### 2.6. Principal Component Analysis of Different Evaluated Traits

_{1}showed the strongest positive correlations with BL, BWI, BWE, FF, and W.S/B. A less significant positive correlation was observed with N.S/B. Despite that the SP trait belongs to the same group, it exhibited a non-significant correlation with PC

_{1}. A modest significant negative correlation was detected between FLS and PC

_{1}. The PC

_{2}was strongly positively correlated with CL and CC. However, a less significant positive correlation was observed with CWI, CWE, and N.B/C.

#### 2.7. Dissimilarity Matrix Analysis among the Population

## 3. Discussion

## 4. Materials and Methods

#### 4.1. Plant Material

#### 4.2. Cluster-Related Traits

(mm)]/[Rachis length (cm) × Pedicel Length (mm)]

#### 4.3. Berry-Related Traits

#### 4.4. Statistical Analysis

## 5. Conclusions

## Supplementary Materials

## Author Contributions

## Funding

## Institutional Review Board Statement

## Informed Consent Statement

## Data Availability Statement

## Acknowledgments

## Conflicts of Interest

## References

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**Figure 1.**Close up view of staminate (male;

**left**), hermaphroditic (perfect;

**center**), and imperfect (female,

**right**) muscadine flowers collected from O24-10-2, Noble, and Onyx genotypes, respectively.

**Figure 2.**(

**A**) Characterization of cluster length (CL) trait among muscadine population (n = 90). The bars represent the mean cluster length (±SD) results from five biological replicates among three years (n = 15). The y-axis refers to the cluster length (cm), and the x-axis represents the muscadine genotypes. Means within columns for the same letter followed by different letters differ significantly by Tukey’s test (p < 0.05). Based on median cluster length (~7.6 cm), the population was divided into two equal groups of long and short clusters. The asterisk refers to the standard commercial colored (red) and bronze (green) cultivars selected as controls in the current study, including Noble, Carlos, Majesty, and Fry. (

**B**) A representative image of muscadine genotypes exhibiting the longest (Granny Val) and shortest (Sugargate) clusters.

**Figure 3.**(

**A**) Characterization of cluster width (CWI) trait among muscadine population (n = 90). The bars represent the mean cluster width (±SD) results from five biological replicates among three years (n = 15). The y-axis refers to the cluster width (cm). Means within columns for the same letter followed by different letters differ significantly by Tukey’s test (p < 0.05). Based on median cluster width (~6.3 cm), 47.8% and 52.2% of the muscadine population produced wide and tight clusters, respectively. Other details as in Figure 2. (

**B**) A representative image of muscadine genotypes exhibiting a maximal (O40-21-1) and a minimal (O15-17-1) cluster width.

**Figure 4.**(

**A**) Characterization of cluster-weight (CWE) trait among muscadine population (n = 90). The bars represent the mean CWE (±SD) results from five biological replicates among three years (n = 15). The y-axis refers to the cluster-weight (g). Means within columns for the same letter followed by different letters differ significantly by Tukey’s test (p < 0.05). Based on median CWE (~88.1 g), the population was divided into two equal groups of heavy and light clusters. Other details as in Figure 2. (

**B**) A representative image of muscadine genotypes exhibiting maximal (Granny Val) and minimal CWE (O15-17-1).

**Figure 5.**(

**A**) Characterization of the number of berries/cluster (N.B/C) trait among muscadine population (n = 90). The bars represent the mean N.B/C (±SD) results from five biological replicates among three years (n = 15). The y-axis refers to the number of berries/cluster. Means within columns for the same letter followed by different letters differ significantly by Tukey’s test (p < 0.05). Based on median N.B/C (~8.7 B/C), the population was divided into two equal groups of high and low number of berries/cluster. Other details as in Figure 2. (

**B**) A representative image of the VM genotypes exhibiting maximal N.B/C (O15-11-1, O15-16-1, and O16-9-2) and the muscadine genotype (Sugargate) showing minimal N.B/C.

**Figure 6.**A representative image demonstrates muscadine-grape cluster structure, including (

**A**) compact clusters, (

**B**) semi-compact clusters, and (

**C**) loose clusters.

**Figure 7.**(

**A**) Characterization of cluster compactness (CC) trait among muscadine population (n = 90). The bars represent the mean CC (±SD) results from five biological replicates among three years (n = 15). The y-axis refers to the cluster compactness levels. Means within columns for the same letter followed by different letters differ significantly by Tukey’s test (p < 0.05). Based on median CC (~12.2), the population was divided into three groups based on the cluster intensity, loose, semi-compact, and compact. Other details as in Figure 2. (

**B**) A representative image of the muscadine genotypes exhibiting minimal (E15-10-1) and maximal (O17-17-1) compactness level.

**Figure 8.**Frequency distribution of the cluster-related traits, including cluster length (

**A**), cluster width (

**B**), cluster weight (

**C**), number of berries/cluster (

**D**), and cluster compactness (

**E**) of the muscadine population (n = 90). The skewness degree and p-value of the Kolmogorov-Smirnov normal distribution test are indicated.

**Figure 9.**(

**A**) Characterization of berry length (BL) trait among muscadine population (n = 90). The bars represent the mean BL (±SD) results from five biological replicates among three years (n = 15). The y-axis refers to the berry length (mm). Means within columns for the same letter followed by different letters differ significantly by Tukey’s test (p < 0.05). Based on median BL (~27.9 mm), the population was divided into two groups of long berries (44 genotypes, 48.9% of the population) and short berries (46 genotypes, 51.1% of the population). Other details are as in Figure 2. (

**B**) A representative image of the muscadine genotypes exhibiting maximal (Supreme) and minimal (Fry Seedless) berry length.

**Figure 10.**(

**A**) Characterization of berry width (BWI) trait among muscadine population (n = 90). The bars represent the mean BWI (±SD) results from five biological replicates among three years (n = 15). The y-axis refers to the berry width (mm). Means within columns for the same letter followed by different letters differ significantly by Tukey’s test (p < 0.05). Based on median BWI (~26.0 mm), the population was divided into two equal groups of wide and tight berries. Other details are as in Figure 2. (

**B**) A representative image of the muscadine genotypes exhibiting maximal (Majesty) and minimal (O15-11-1) berry width.

**Figure 11.**Characterization of berry weight (BWE) trait among muscadine population (n = 90). The bars represent the mean BWE (±SD) results from five biological replicates among three years (n = 15). The y-axis refers to the berry weight (g). Means within columns for the same letter followed by different letters differ significantly by Tukey’s test (p < 0.05). Based on median BWE (~11.9 g), the population was divided into two equal groups of heavy and light berries. Other details are as in Figure 2. A representative image for the muscadine genotypes exhibiting maximal and minimal berry weight is as in Figure 10.

**Figure 12.**(

**A**) Characterization of number of seeds/berry (N.S/B) trait among muscadine population (n = 90). The bars represent the mean N.S/B (±SD) results from five biological replicates among three years (n = 15). The y-axis refers to the number of seeds/berry. Means within columns for the same letter followed by different letters differ significantly by Tukey’s test (p < 0.05). Based on median N.S/B (~3.6), the population was divided into two equal groups of a high and low seed number. Other details are as in Figure 2. (

**B**) A representative image of the muscadine genotypes exhibiting maximal (Onyx) and minimal (Fry Seedless) number of seeds/berry.

**Figure 13.**Characterization of the weight of seeds/berry (W.S/B) trait among muscadine population (n = 90). The bars represent the mean W.S/B (±SD) results from five biological replicates among three years (n = 15). The y-axis refers to the weight of seeds/berry (g). Means within columns for the same letter followed by different letters differ significantly by Tukey’s test (p < 0.05). Based on median W.S/B (~0.34 g), the population was divided into two equal groups of heavy and light seed weight. Other details are as in Figure 2. A representative image of the muscadine genotypes exhibiting maximal and minimal weight of seeds/berry is as in Figure 12.

**Figure 14.**(

**A**) Characterization of firmness (FF) trait among muscadine population (n = 90). The bars represent the mean FF (±SD) results from five biological replicates among three years (n = 15). The y-axis refers to the firmness (N). Means within columns for the same letter followed by different letters differ significantly by Tukey’s test (p < 0.05). The population was divided into a firm, mid-firm, and soft berries based on median FF (~0.11 N). Other details are as in Figure 2. (

**B**) A representative image of the muscadine genotypes exhibiting maximal (A14-13-1) and minimal (C11-2-2) berry firmness.

**Figure 15.**(

**A**) Characterization of scar pattern (SP) trait among muscadine population (n = 90). The bars represent the mean SP (±SD) results from five biological replicates among three years (n = 15). The y-axis refers to the dry scar pattern (%). Means within columns for the same letter followed by different letters differ significantly by Tukey’s test (p < 0.05). Based on median SP (~48%), the population was divided into two equal groups of dry scar and wet scar. Other details are as in Figure 2. A close-up view of muscadine berries showing dry scar vs. wet scar patterns in muscadine berries is indicated. (

**B**) A representative image of the muscadine genotypes exhibiting extreme dry (O28-9-2) and wet (O18-2-1) berry scar.

**Figure 16.**Frequency distribution of the berry-related traits, including berry length (

**A**), berry width (

**B**), berry weight (

**C**), number of seeds/berry (

**D**), the weight of seeds/berry (

**E**), firmness (

**F**), and scar pattern (

**G**) of the muscadine population (n = 90). The skewness degree and p-value of the Kolmogorov-Smirnov normal distribution test are indicated.

**Figure 17.**Hierarchical clustering of the different evaluated traits of muscadine population (n = 90). Data of different traits, including variables of cluster length (CL), cluster width (CWI), cluster weight (CWE), number of berries/cluster (N.B/C), cluster compactness (CC), berry length (BL), berry width (BWI), berry weight (BWE), number of seeds/berry (N.S/B), the weight of seeds/berry (W.S/B), firmness (FF), and scar pattern (SP) are presented as an average of five biological replicates among three years (n = 15). The log2-transformed values of each character are represented by colors. Green and red boxes indicate higher and lower values, respectively. The color change is proportional to the two extremes (see the color scale at the top of the figure).

**Figure 18.**Principal Component Analysis (PCA) scatter plots of the different cluster- and berry-related traits, including variables of cluster length (CL), cluster width (CWI), cluster weight (CWE), number of berries/cluster (N.B/C), cluster compactness (CC), berry length (BL), berry width (BWI), berry weight (BWE), number of seeds/berry (N.S/B), the weight of seeds/berry (W.S/B), firmness (FF), and scar pattern (SP). The flower structure (FLS) trait was also covered in the analysis. The scatter was generated using the average of three biological replicates. According to the PCA model, 40.53% and 20.91% of the variance were explained by the first principle component (PC

_{1}) and the second principle component (PC

_{2}), respectively.

**Figure 19.**Dissimilarity matrix showing the distances among the genotypes. The gradient of color indicates the distance between genotypes; the green color denotes the highest dissimilarity, and the pink color means the lowest genetic distance. In addition, the pink color represents the diagonal.

**Figure 20.**A representative image of leaves, shoots, and cluster of promising muscadine advanced selection genotypes suitable for white (C8-6-1 and O44-14-1) and red (B20-18-2 and C11-2-2) wine production. The standard cultivars Carlos and Noble were used as controls.

**Table 1.**Cluster characteristics of muscadine genotypes with high performance to promote into new standard cultivars.

Genotype | Flower | Cluster Wgt. (g) | No. Berries/Cluster | Cluster Compactness | Yield (kg) |
---|---|---|---|---|---|

Carlos * | Perfect | 41.9 ± 1.4 | 7.6 ± 0.6 | 6.8 ± 0.8 | 5.4 ± 0.8 |

C8-6-1 | Female | 149.1 ± 3.1 | 20.8 ± 2.7 | 28.9 ± 1.5 | 17.3 ± 1.5 |

O44-14-1 | Perfect | 56.4 ± 0.6 | 13.8 ± 0.4 | 11.4 ± 1.1 | 9.9 ± 1.0 |

Noble * | Perfect | 47.8 ± 1.5 | 12.2 ± 0.4 | 6.9 ± 0.8 | 3.9 ± 0.4 |

B20-18-2 | Perfect | 61.1 ± 1.1 | 13.2 ± 1.1 | 9.2 ± 1.0 | 11.2 ± 1.2 |

C11-2-2 | Perfect | 97.9 ± 5.7 | 27.6 ± 4.7 | 18.9 ± 1.3 | 11.3 ± 1.6 |

Average ** | - | 96.9 ± 42.7 | 11.1 ± 6.8 | 13.2 ± 6.1 | 6.6 ± 4.5 |

Median | - | 88.1 | 8.7 | 12.2 | 5.3 |

**Table 2.**Berry characteristics of muscadine genotypes with high performance to promote into new standard cultivars.

Genotype | Color | Berry Wgt. (g) | Dry Scar (%) | Firmness (N) | No. Seeds/Berry | Wgt. Seeds/Berry (g) |
---|---|---|---|---|---|---|

Carlos * | Bronze | 6.7 ± 0.4 | 54 ± 1.1 | 0.16 ± 0.04 | 4.0 ± 0 | 0.34 ± 0.05 |

C8-6-1 | Bronze | 8.6 ± 0.5 | 18 ± 0.9 | 0.10 ± 0 | 4.0 ± 0 | 0.44 ± 0.05 |

O44-14-1 | Bronze | 4.6 ± 0.5 | 72 ± 2.9 | 0.05 ± 0 | 3.2 ± 0.4 | 0.22 ± 0.04 |

Noble * | Black | 4.2 ± 0.3 | 28 ± 1.1 | 0.06 ± 0.02 | 3.8 ± 0.4 | 0.24 ± 0.05 |

B20-18-2 | Black | 5.2 ± 0.3 | 18 ± 0.7 | 0.10 ± 0 | 4.2 ± 0.4 | 0.27 ± 0.03 |

C11-2-2 | Dark red | 3.0 ± 0.3 | 64 ± 2.6 | 0.03 ± 0 | 3.2 ± 0.4 | 0.28 ± 0.04 |

Average ** | - | 12.2 ± 5.6 | 49.7 ± 22.4 | 0.12 ± 0.05 | 3.6 ± 0.7 | 0.35 ± 0.11 |

Median | - | 11.9 | 48 | 0.11 | 3.6 | 0.34 |

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**MDPI and ACS Style**

Campbell, J.; Sarkhosh, A.; Habibi, F.; Ismail, A.; Gajjar, P.; Zhongbo, R.; Tsolova, V.; El-Sharkawy, I.
Biometrics Assessment of Cluster- and Berry-Related Traits of Muscadine Grape Population. *Plants* **2021**, *10*, 1067.
https://doi.org/10.3390/plants10061067

**AMA Style**

Campbell J, Sarkhosh A, Habibi F, Ismail A, Gajjar P, Zhongbo R, Tsolova V, El-Sharkawy I.
Biometrics Assessment of Cluster- and Berry-Related Traits of Muscadine Grape Population. *Plants*. 2021; 10(6):1067.
https://doi.org/10.3390/plants10061067

**Chicago/Turabian Style**

Campbell, Jiovan, Ali Sarkhosh, Fariborz Habibi, Ahmed Ismail, Pranavkumar Gajjar, Ren Zhongbo, Violeta Tsolova, and Islam El-Sharkawy.
2021. "Biometrics Assessment of Cluster- and Berry-Related Traits of Muscadine Grape Population" *Plants* 10, no. 6: 1067.
https://doi.org/10.3390/plants10061067