## 3 Results
### Metabolomics Analysis of Three Raspberry Cultivars Leaves
\begin{table}
\begin{tabular}{c c c c c c c} \hline
**Up-Regulated Metabolites** & **VIP** & **log2(FC)** & _p._****Ajusted** & **Down-Regulated Metabolites** & **VIP** & **log2(FC)** & _p._**Ajusted** \\ \hline \multicolumn{7}{c}{(1R,6R)-6-Hydroxy-2-} \\ Castosterone & 3.533 & 8.7869 & 0.000019 & succinylcyclohex-2,4-diene- & 2.2308 & \(-\)7.1632 & 0.015585 \\ \multicolumn{7}{c}{1-carboxylate} \\ Propinol adenylate & 2.316 & 6.9795 & 0.032560 & Phenylacetaldehyde & 1.826 & \(-\)5.8687 & 0.000909 \\ Pantetheine 4\({}^{\prime}\)-phosphate & 2.2522 & 5.545 & 0.000002 & Adenosine phosphosulfate & 1.7621 & \(-\)4.6282 & 0.000007 \\
4-Hydroxycinnamic acid & 2.2308 & 5.3486 & 0.000000005 & SAICAR & 1.6775 & \(-\)4.57 & 0.017379 \\ \multicolumn{7}{c}{6-Germany/jeranyl-2-} \\ \multicolumn{7}{c}{methylbenzene-1,4-diol} \\ Kaur-16-en-18-oic acid & 2.0679 & 5.0538 & 0.000150 & Prostaglandin H2 & 1.2666 & \(-\)3.282 & 0.016800 \\ Behenic acid & 1.9624 & 4.769 & 0.000002 & 15(S)-HPETE & 1.2294 & \(-\)3.1716 & 0.005833 \\ \hline \end{tabular}
\end{table} 
Table 3: Differential accumulated metabolites top 10 (Autumn Bliss vs. Red Autumn, negative ion mode). \begin{table}
\begin{tabular}{c c c c c c c c} \hline
**Up-Regulated Metabolites** & **VIP** & **log2(FC)** & _p._**Ajusted** & **Down-Regulated Metabolites** & **VIP** & **log2(FC)** & _p._**Ajusted** \\ \hline \multicolumn{7}{c}{(1R,6R)-6-Hydroxy-2-} \\ Castosterone & 3.533 & 8.7869 & 0.000019 & succinylcyclohex-2,4-diene- & 2.2308 & \(-\)7.1632 & 0.015585 \\ \multicolumn{7}{c}{1-carboxylate} \\ Propinol adenylate & 2.316 & 6.9795 & 0.032560 & Phenylacetaldehyde & 1.826 & \(-\)5.8687 & 0.000909 \\ Pantetheine 4\({}^{\prime}\)-phosphate & 2.2522 & 5.545 & 0.000002 & Adenosine phosphosulfate & 1.7621 & \(-\)4.6282 & 0.0000007 \\
4-Hydroxycinnamic acid & 2.2308 & 5.3486 & 0.000000005 & SAICAR & 1.6775 & \(-\)4.57 & 0.017379 \\ \multicolumn{7}{c}{6-Germany/jeranyl-2-} \\ \multicolumn{7}{c}{methylbenzene-1,4-diol} \\ Kaur-16-en-18-oic acid & 2.0679 & 5.0538 & 0.000150 & Prostaglandin H2 & 1.2666 & \(-\)3.282 & 0.016800 \\ Behenic acid & 1.9624 & 4.769 & 0.000002 & 15(S)-HPETE & 1.2294 & \(-\)3.1716 & 0.005833 \\ \hline \end{tabular}
\end{table} 
Table 1: Differential accumulated metabolites top 10 (Autumn vs. Autumn Bliss, negative ion mode). In order to further distinguish the differentially accumulated metabolites between different cultivars, comparisons between each two different cultivars were employed to identify up or down regulated metabolites. Then the differentially accumulated metabolites in different comparisons were grouped, and the selected differential metabolites in each group were analyzed for KEGG pathway enrichment. The comparison of Autumn Britten vs. Autumn Bliss found 36 up-regulated metabolites enriched in 38 metabolic pathways, and 65 down-regulated differential metabolites enriched in 46 metabolic pathways; Autumn Britten vs. Red Autumn found 48 up-regulated metabolites enriched in 44 metabolic pathways, and 108 down-regulated metabolites enriched in 55 metabolic pathways; Autumn Bliss vs. Red Autumn found 88 up-regulated metabolites enriched in 50 metabolic pathways (Figure 3). Figure 2: Volcano plots of metabolites detected in cultivars for different comparisons: (**A**) Autumn Bliss vs. Autumn Britten; (**B**) Autumn Bliss vs. Red Autumn; (**C**) Autumn Britten vs. Red Autumn; in negative ion mode: (**D**) Autumn Bliss vs. Autumn Britten; (**E**) Autumn Bliss vs. Red Autumn; (**F**) Autumn Britten vs. Red Autumn; in positive ion mode. The vertical dotted lines indicate fold change of 1.5, and the horizontal dotted line indicates \(p\) = 0.05. Figure 3: Metabolic pathway analysis of different comparisons of three cultivars. (**A**) Autumn Bliss vs. Autumn Britten (up-regulation metabolites); (**B**) Autumn Britten vs. Red Autumn (up-regulation metabolites); (**C**) Autumn Bliss vs. Red Autumn (up-regulation metabolites); (**D**) Autumn Bliss vs.