为揭示同时导入多个抗性基因对受体植物库安托杨代谢物层面的影响机制，我所遗传育种研究创新团队利用气相色谱-质谱联用仪（GC-MS）和超高效液相色谱-串联质谱（UPLC-MS/MS）技术，从代谢组水平研究了2个转多基因（Vgb、SACB、JERF36、BtCry3A和Oc-I）杨树新品种（多抗杨2号和多抗杨3号）和1个非转基因（D5-0）形成层代谢产物，检测鉴定转多基因与非转基因杨树代谢物种类和丰度变化差异，以期能够找到与转入目的基因相关的标记代谢物或与响应外界非生物胁迫目标性状关联的代谢物。研究共鉴定获得了239个命名的代谢物，其中197种代谢物在三个不同株系间差异显著，这些代谢产物占据了9条初级代谢途径和44条次级代谢途径。精氨酸和谷氨酸作为氮代谢的底物和中间产物在生长和胁迫相关过程中发挥着重要作用，细胞壁前体物质蔗糖、尿苷二磷酸葡萄糖及其衍生物和与转基因杨树抗虫性相关的邻苯二酚，这些代谢物在转基因品系中水平更高，且与非转基因杨树之间的差异较为显著。相关研究结果为进一步研究杨树形成层代谢奠定基础，并为鉴定获得与应激响应相关的标记代谢物提供参考。本研究是首次对转基因杨树进行代谢组水平研究，阐明多基因表达促进杨树生长及抗性增强的内在机制，鉴定了与靶基因及非预期性状相关代谢物，从代谢组层面扩展了转基因林木评价指标体系。本研究结果在SCI收录期刊Frontiers in Plant Science上发表。（影响因子：3.677，分区：生物2区；林业所：宁坤、丁昌俊、苏晓华等）
Figure 1.
Distribution of identified metabolites in the cambium of non-transgenic and
transgenic poplar. (A) Heat
map showing the clustering of detected metabolites into nine classes across the
three lines. Different colors represent nine metabolic pathways of the 239
metabolites, and the numerals in parentheses represent the numbers of
metabolites identified in the primary metabolic pathways. (B) The
distribution of metabolic pathways of the differential 197 metabolites.
Ordinate reports the various metabolic pathways, while abscissa represents the
number of metabolites. Different colors represent primary pathways.
Figure 2.
PCA score plots of metabolome data from the cambium of transgenic and
non-transgenic poplar. PCA score
plot of the three lines. The red circle indicates non-transgenic line D5-0,
while the green square and blue triangle indicate multi-gene transgenic line
D5-20 and D5-21, respectively. The plot displays PC1 on the ordinate axis and
PC2 on the abscissa.
Figure 3. Four most important compounds and top 30 metabolites. (A) The four most important compounds for correct classification
of MDA value >10 from non-transgenic and transgenic polar. The ordinates
represent scaled intensity. (B) The top 30 metabolites for classification.
Figure 4.
The ten most variable compounds. (A) The ratio between maximum and minimum mean
values for the most variable compounds by variety (>3 folds mean range). (B)
The relative abundance of the ten variable compounds. The ordinates represent
scaled intensity of the relative abundance. Error bars are shown for six
replicates of the relative abundance. Different letters indicate a
statistically significant difference when analyzed by one-way ANOVA and a
multiple comparison using Duncan’s test at p ≤ 0.05. For column diagram with no
standards deviations, eicosenoate (20:1n9 or 11), 13-HODE + 9-HODE and
12,13-DiHOME in D5-0 line, the standards deviations are not shown due to the
large variation in the presence of one to two biological replicates and the
standards deviations are greater than the average value.
Figure 5.
Metabolites associated with cell wall and polysaccharide metabolisms. The
ordinates represent scaled intensity of the relative abundance. Error bars are
shown for six replicates of the relative abundance. Different letters indicate
a statistically significant difference when analyzed by one-way ANOVA and a
multiple comparison using Duncan’s test at p ≤ 0.05.